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Sample records for chemically-induced rat mesenchymal

  1. Altered magnesium transport in slices of kidney cortex from chemically-induced diabetic rats

    Energy Technology Data Exchange (ETDEWEB)

    Hoskins, B.

    1981-10-01

    The uptake of magnesium-28 was measured in slices of kidney cortex from rats with alloxan-diabetes and from rats with streptozotocin-diabetes of increasing durations. In both forms of chemically-induced diabetes, magnesium-28 uptake by kidney cortex slices was significantly increased over uptake measured in kidney cortex slices from control rats. Immediate institution of daily insulin therapy to the diabetic rats prevented the diabetes-induced elevated uptake of magnesium without controlling blood glucose levels. Late institution of daily insulin therapy was ineffective in restoring the magnesium uptake to control values. These alterations in magnesium uptake occurred prior to any evidence of nephropathy (via the classic indices of proteinuria and increased BUN levels). The implications of these findings, together with our earlier demonstrations of altered calcium transport by kidney cortex slices from chemically-induced diabetic rats, are discussed in terms of disordered divalent cation transport being at least part of the basic pathogenesis underlying diabetic nephropathy.

  2. Absence of p53 gene expression in selenium molecular prevention of chemically Induced hepatocarcinogenesis in rats

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    Nasar Y Alwahaibi

    2011-01-01

    Full Text Available Background/Aim: p53 pathway is thought by many researchers to be critically involved in selenium′s chemoprevention or in hepatocarcinogenesis. The aim of this study was to investigate the gene expression of p53, p21 and B-cell lymphoma-2 (bcl-2 using preventive and therapeutic approaches of selenium in chemically induced hepatocarcinogenesis in rats. Materials and Methods: Rats were divided randomly into six groups: Negative control, positive control (diethyl nitrosamine +2-acetylaminofluorene, preventive group, preventive control (respective control for preventive group, therapeutic group and therapeutic control (respective control for therapeutic group. p53, p21 and bcl-2 genes on liver tissues were measured using real-time polymerase chain reaction. Results: The expression of p53 was only significant in the therapeutic control. The expression of bcl-2 was insignificant in all the groups. p21 expression was significant in all the groups except the preventive group. Conclusions: The selenium molecular mechanism for liver cancer prevention is not through the p53 pathway. Also, the absence of p53 is not necessary for chemically induced liver cancer in rats.

  3. Antiinflammatory effects of human milk on chemically induced colitis in rats.

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    Grazioso, C F; Werner, A L; Alling, D W; Bishop, P R; Buescher, E S

    1997-11-01

    We examined the effects of a human milk diet on rats with chemical colitis induced with a 4% acetic acid enema. Colonic myeloperoxidase activity was used as a surrogate marker for neutrophil infiltration. Control rats fed rat chow had little colonic myeloperoxidase activity; geometric mean, 0.27 U/g of tissue. Rats with colitis fed rat chow had significantly increased colonic myeloperoxidase activity (geometric mean, 6.76 U/g, p Pedialyte (geometric mean, 6.92 and 8.13 U/g, respectively, both p < 0.01 versus no colitis). Animals with colitis fed human milk had significantly lower colonic myeloperoxidase activity (geometric mean, 2.34 U/g) than did animals with colitis fed either chow or infant formula (p < 0.001). Similar effects were seen in rats with colitis fed infant formula supplemented with recombinant human IL-1 receptor antagonist (geometric mean, 1.95 U/g). These data show that orally administered human milk has an antiinflammatory effect on chemically induced colitis in rats, which may be mediated in part by IL-1 receptor antagonist contained in human milk.

  4. The relationship between chemical-induced kidney weight increases and kidney histopathology in rats.

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    Craig, Evisabel A; Yan, Zhongyu; Zhao, Q Jay

    2015-07-01

    The kidney is a major site of chemical excretion, which results in its propensity to exhibit chemically-induced toxicological effects at a higher rate than most other organs. Although the kidneys are often weighed in animal toxicity studies, the manner in which these kidney weight measurements are interpreted and the value of this information in predicting renal damage remains controversial. In this study we sought to determine whether a relationship exists between chemically-induced kidney weight changes and renal histopathological alterations. We also examined the relative utility of absolute and relative (kidney-to-body weight ratio) kidney weight in the prediction of renal toxicity. For this, data extracted from oral chemical exposure studies in rats performed by the National Toxicology Program were qualitatively and quantitatively evaluated. Our analysis showed a statistically significant correlation between absolute, but not relative, kidney weight and renal histopathology in chemically-treated rats. This positive correlation between absolute kidney weight and histopathology was observed even with compounds that statistically decreased terminal body weight. Also, changes in absolute kidney weight, which occurred at subchronic exposures, were able to predict the presence or absence of kidney histopathology at both subchronic and chronic exposures. Furthermore, most increases in absolute kidney weight reaching statistical significance (irrespective of the magnitude of change) were found to be relevant for the prediction of histopathological changes. Hence, our findings demonstrate that the evaluation of absolute kidney weight is a useful method for identifying potential renal toxicants.

  5. Bromophenacyl bromide, a phospholipase A2 inhibitor attenuates chemically induced gastroduodenal ulcers in rats

    Institute of Scientific and Technical Information of China (English)

    Mohammad Tariq; Ibrahim Elfaki; Haseeb Ahmad Khan; Mohammad Arshaduddin; Samia Sobki; Meshal Al Moutaery

    2006-01-01

    AIM: To study the effect of bromophenacyl bromide (BPB), a phospholipase A2 inhibitor on gastric secretion and to protect chemically induced gastric and duodenal ulcers in rats.METHODS: Acid secretion studies were undertaken in pylorus-ligated rats with BPB treatment (0, 5, 15 and 45 mg/kg). Gastric and duodenal lesions in the rats were induced by ethanol and cysteamine respectively. The levels of gastric wall mucus, nonprotein sulfhydryls (NPSH) and myeloperoxidase (MPO) were also measured in the glandular stomach of rats following ethanol induced gastric lesions.RESULTS: BPB produced a dose-dependent inhibition of gastric acid secretion and acidity in rats. Pretreatment with BPB significantly attenuated the formation of ethanol induced gastric lesion. BPB also protected intestinal mucosa against cysteamine-induced duodenal ulcers.The antiulcer activity of BPB was associated with significant inhibition of ethanol-induced depletion of gastric wall mucus, NP-SH and MPO. These findings pointed towards the mediation of sulfhydryls in BPB induced gastrointestinal cytoprotection.CONCLUSION: BPB possesses significant antiulcer and cytoprotective activity against experimentally induced gastroduodenal lesions.

  6. Protective role of food supplement Spirulina fusiformis in chemical induced hepatotoxicity: A Bromobenzene model in rats

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    Evan Prince Sabina

    2014-03-01

    Full Text Available The present study evaluated the efficacy of Spirulina fusiformis in protecting against chemical induced hepatotoxicity in rats using Bromobenzene as the candidate toxin. A single oral dose of bromobenzene (BB (10mmol/kg b.w. resulted in significant (p< 0.05 decrease in antioxidant levels (catalase, superoxide dismutase, glutathione-S-transferase, glutathione peroxidese, total reduced glutathione and total protein, and significant (p< 0.05 increase in the levels of serum bilirubin, liver enzymes (alanine transaminase, aspartate transaminase and alkaline phosphatase indicating the induction of hepatotoxicity. Spirulina fusiformis (400 mg/kg b.w was orally administered for 8 days prior to the administration of BB and was seen to protect the above parameters from significant changes upon challenge with bromobenzene. This was also confirmed by the histological examination of liver tissues after sacrifice. The protective effect of Spirulina fusiformis was comparable to that of the standard hepatoprotective drug sylimarin.

  7. Comparative sensitivity of human and rat neural cultures to chemical-induced inhibition of neurite outgrowth

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    Harrill, Joshua A.; Freudenrich, Theresa M.; Robinette, Brian L.; Mundy, William R., E-mail: mundy.william@epa.gov

    2011-11-15

    cultures were more sensitive to neurite outgrowth inhibitors, they also had a lower dynamic range for detecting chemical-induced neurite outgrowth inhibition and greater variability from culture-to-culture as compared to rat primary cortical cultures.

  8. In vivo photo-detection of chemically induced premalignant lesions and squamous cell carcinoma of the rat palatal mucosa

    NARCIS (Netherlands)

    Nauta, JM; Speelman, OC; vanLeengoed, HLLM; Nikkels, PGJ; Roodenburg, JLN; Witjes, MJH; Vermey, A

    1997-01-01

    Photo-detection using in vivo fluorescence was studied for different stages of chemically induced premalignant lesions and squamous cell carcinoma (SCC) of the Wistar rat palatal mucosa. It was found that the epithelial dysplasia (numerically expressed in the epithelial atypia index (EAI)) of the ra

  9. Use of a chemically induced-colon carcinogenesis-prone Apc-mutant rat in a chemotherapeutic bioassay

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    Yoshimi Kazuto

    2012-10-01

    Full Text Available Abstract Background Chemotherapeutic bioassay for colorectal cancer (CRC with a rat model bearing chemically-induced CRCs plays an important role in the development of new anti-tumor drugs and regimens. Although several protocols to induce CRCs have been developed, the incidence and number of CRCs are not much enough for the efficient bioassay. Recently, we established the very efficient system to induce CRCs with a chemically induced-colon carcinogenesis-prone Apc-mutant rat, Kyoto Apc Delta (KAD rat. Here, we applied the KAD rat to the chemotherapeutic bioassay for CRC and showed the utility of the KAD rat. Methods The KAD rat has been developed by the ENU mutagenesis and carries a homozygous nonsense mutation in the Apc gene (S2523X. Male KAD rats were given a single subcutaneous injection of AOM (20 mg/kg body weight at 5 weeks of age. Starting at 1 week after the AOM injection, they were given 2% DSS in drinking water for 7 days. Tumor-bearing KAD rats were divided into experimental and control groups on the basis of the number of tumors observed by endoscopy at week 8. The 5-fluorouracil (5-FU was administrated intravenously a dose of 50 or 75 mg/kg weekly at week 9, 10, and 11. After one-week interval, the 5-FU was given again at week 13, 14, and 15. At week 16, animals were sacrificed and tumor number and volume were measured macroscopically and microscopically. Results In total 48 tumors were observed in 27 KAD rats with a 100% incidence at week 8. The maximum tolerated dose for the KAD rat was 50 mg/kg of 5-FU. Macroscopically, the number or volume of tumors in the 5-FU treated rats was not significantly different from the control. Microscopically, the number of adenocarcinoma in the 5-FU treated rats was not significantly different (p Conclusion The use of the AOM/DSS-treated tumor-bearing KAD rats could shorten the experimental period and reduce the number of animals examined in the chemotherapeutic bioassay. The

  10. Detection and characterization of chemical-induced abnormal tissue and rat tumors at different stages using fluorescence spectroscopy

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    Chen, Wei R.; Jassemnejad, Baha; Crull, Jason; Knobbe, Edward T.; Nordquist, Robert E.

    1996-04-01

    Fluorescence spectroscopy of diseased tissues, including chemical-induced rat liver, kidney and testis lesions, as well as murine mammary tumor, was studied. The rat liver, kidney and testis tissues were excited by radiation of 350 and 366 nm, which appeared to provide the optimal differentiation between normal and lesion tissues; the tumor tissues were excited by both 350 nm and 775 nm wavelengths. In comparison with normal liver tissue, at (lambda) ex equals 366 nm, the fluorescent spectrum of liver lesion showed a clear red shift around the emission peak of 470 nm, the major native fluorescent peak of organized tissue. When excited by 350 nm wavelength, all the chemically induced lesion tissues (liver, kidney and testis) appeared to cause a significant reduction of emission intensity at the 470 nm peak. While the 775 nm excitation did not reveal any significant difference among tumor, muscle and skin tissues, the 350 nm excitation did provide some interesting features among the tumor tissues at different stages. Compared with muscle tissue, the viable tumor showed an overall reduction of emission intensity around 470 nm. In addition, the viable tumor tissue showed a secondary emission peak at 390 nm with necrotic tumor tissue having a reduced intensity. The histology of both viable and necrotic tumor tissue was examined and appeared to correlate with the results of the fluorescent spectroscopy observation.

  11. Chemically induced Salmonella enteritidis ghosts as a novel vaccine candidate against virulent challenge in a rat model.

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    Vinod, Nagarajan; Oh, Sung; Kim, Seongdae; Choi, Chang Won; Kim, Sei Chang; Jung, Cheong-Hwan

    2014-05-30

    Salmonella enteritidis ghosts (SEGs), non-living empty bacterial cell envelopes were generated by using the minimum inhibitory concentration (MIC) of sodium hydroxide (NaOH) and investigated as a vaccine candidate in rats. To determine the immunogenicity and protective efficacy of SEG vaccine, rats were divided into four groups: group A (non-vaccinated control), group B (orally vaccinated), group C (intramuscularly vaccinated) and group D (intramuscularly vaccinated with complete Freund's adjuvant). Vaccination of rats with SEGs induced significant immune responses before and after virulent challenge. Rats vaccinated with SEGs showed significant increases in serum IgG antibodies after challenging with virulent S. enteritidis on week 8 and week 10 (P<0.01). During the vaccination period, groups B, C and D showed significantly higher serum bactericidal activity (SBA) compared to group A (P<0.01). Most importantly, bacterial loads in vaccinated groups were significantly lower than in the non-vaccinated group (P<0.01). In conclusion, these results show that the chemically induced SEGs as a vaccine candidate against virulent challenge.

  12. Saffron Aqueous Extract Inhibits the Chemically-induced Gastric Cancer Progression in the Wistar Albino Rat

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    S. Zahra Bathaie

    2013-01-01

    Full Text Available Objective(s: Gastric cancer is the first and second leading cause of cancer related death in Iranian men and women, respectively. Gastric cancer management is based on the surgery, radiotherapy and chemotherapy. In the present study, for the first time, the beneficial effect of saffron (Crocus sativus L. aqueous extract (SAE on the 1-Methyl-3-nitro-1-nitrosoguanidine (MNNG-induced gastric cancer in rat was investigated. Materials and Methods: MNNG was used to induce gastric cancer and then, different concentrations of SAE were administered to rats. After sacrificing, the stomach tissue was investigated by both pathologist and flow cytometry, and several biochemical parameters was determined in the plasma (or serum and stomach of rats. Results: Pathologic data indicated the induction of cancer at different stages from hyperplasia to adenoma in rats; and the inhibition of cancer progression in the gastric tissue by SAE administration; so that, 20% of cancerous rats treated with higher doses of SAE was completely normal at the end of experiment and there was no rat with adenoma in the SAE treated groups. In addition, the results of the flow cytometry/ propidium iodide staining showed that the apoptosis/proliferation ratio was increased due to the SAE treatment of cancerous rats. Moreover, the significantly increased serum LDH and decreased plasma antioxidant activity due to cancer induction fell backwards after treatment of rats with SAE. But changes in the other parameters (Ca2+, tyrosine kinase activity and carcino-embryonic antigen were not significant. Conclusion: SAE inhibits the progression of gastric cancer in rats, in a dose dependent manner.

  13. Aqueous suspension of anise “Pimpinella anisum” protects rats against chemically induced gastric ulcers

    OpenAIRE

    2007-01-01

    AIM: To substantiate the claims of Unani and Arabian traditional medicine practitioners on the gastroprotective potential effect of a popular spice anise, “Pimpinella anisum L.” on experimentally-induced gastric ulceration and secretion in rats.

  14. Effect of Cu supplementation on genomic instability in chemically-induced mammary carcinogenesis in the rat

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    Bobrowska Barbara

    2011-12-01

    Full Text Available Abstract Backround The aim of the present study was to assess the effect of dietary supplementation (copper or copper and resveratrol on the intensity of carcinogenesis and the frequency of microsatellite instability in a widely used model of mammary carcinogenesis induced in the rat by treatment with 7,12-dimethylbenz[a]anthracene (DMBA. Methods DNA was extracted from rat mammary cancers and normal tisues, amplified by PCR, using different polymorphic DNA markers and the reaction products were analyzed for microsatellite instability. Results It was found that irrespectively of the applied diet there was no inhibition of mammary carcinogenesis in the rats due to DMBA. Besides, in the groups supplemented with Cu (II or Cu (II and resveratrol the tumor formation was clearly accelerated. Unlike the animals that were fed with standard diet, the supplemented rats were characterized by the loss of heterozygosity of microsatellite D3Mgh9 in cancer tumors (by respectively 50 and 40%. When the animals received Cu (II and resveratrol supplemented diet the occurrence of genomic instability was additionally found in their livers in the case of microsatellite D1Mgh6 (which was stable in the animals without dietary supplementation. Conclusions Identification of the underlying mechanisms by which dietary factors affect genomic stability might prove useful in the treatment of mammary cancer as well as in the incorporation of dietary factors into mammary cancer prevention strategies.

  15. Aqueous suspension of anise "Pimpinella anisum" protects rats against chemically induced gastric ulcers

    Institute of Scientific and Technical Information of China (English)

    Ibrahim A Al Mofleh; Abdulqader A Alhaider; Jaber S Mossa; Mohammed O Al-Soohaibani; Syed Rafatullah

    2007-01-01

    AIM:To substantiate the claims of Unani and Arabian traditional medicine practitioners on the gastroprotective potential effect of a popular spice anise,"Pimpinella anisum L." on experimentally-induced gastric ulceration and secretion in rats.METHODS:Acute gastric ulceration in rats was produced by various noxious chemicals including 80% ethanol,0.2 mol/L NaOH,25% NaCl and indomethacin.Anti-secretory studies were undertaken using pylorusligated Shay rat technique.Levels of gastric non-protein sulfhydryls(NP-SH)and wall mucus were estimated and gastric tissue was also examined histologically.Anise aqueous suspension was used in two doses(250 and 500 mg/kg body weight)in all experiments.RESULTS:Anise significantly inhibited gastric mucosal damage induced by necrotizing agents and indomethacin.The anti-ulcer effect was further confirmed histologically.In pylorus-ligated Shay rats,anise suspension significantly reduced the basal gastric acid secretion,acidity and completely inhibited the rumenal ulceration.On the other hand,the suspension significantly replenished ethanol-induced depleted levels of gastric mucosal NP-SH and gastric wall mucus concentration.CONCLUSION:Anise aqueous suspension possesses significant cytoprotective and anti-ulcer activities against experimentally-induced gastric lesions.The anti-ulcer effect of anise is possibly prostaglandin-mediated and/or through its anti-secretory and antioxidative properties.

  16. Effect of chronic administration of green tea extract on chemically induced electrocardiographic and biochemical changes in rat heart

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    Patil Leena

    2010-01-01

    Full Text Available Many chemicals induce cell-specific cytotoxicity. Chemicals like doxorubicin induce oxidative stress leading to cardiotoxicity causing abnormalities in ECG and increase in the biomarkers indicating toxicity. Green tea extract (GTE, Camellia sinensis (Theaceae, is reported to exert antioxidant activity mainly by means of its polyphenolic constituent, catechins. Our study was aimed to find out the effect of GTE (25, 50, 100 mg/kg/day p.o. for 30 days on doxorubicin-induced (3 mg/kg/week, i.p. for 5 weeks electrocardiographic and biochemical changes in rat heart. It is observed that GTE administered rats were less susceptible to doxorubicin-induced electrocardiographic changes and changes in biochemical markers like lactate dehydrogenase (LDH, creatine kinase (CK, and glutamic oxaloacetate transaminase (GOT in serum, and superoxide dismutase (SOD, catalase (CAT and reduced glutathione (GSH, membrane bound enzymes like Na + K + ATPase, Ca 2+ ATPase, Mg 2+ ATPase and decreased lipid peroxidation (LP in heart tissue, indicating the protection afforded by GTE administration.

  17. Puerarin exerts the protective effect against chemical induced dysmetabolism in rats.

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    Li, Rong; Song, Jianhua; Wu, Wei; Wu, Xinmou; Su, Min

    2016-12-31

    In this study, we aim to explore the potential benefits of puerarin on metabolic function of liver fibrosis (LF) rat induced by carbon tetrachloride (CCl4), and to investigate with the underlying molecular mechanism targeted on liver and pancreas tissues. In methodology, The LF rats were prepared through intragastrically giving CCl4 twice each week (2ml/kg, v/w) for 8weeks, and dosed puerarin (20, 40mg/kg) were given three times each week via intraperitoneal injection. After being conducted with oral glucose tolerance test (OGTT), the blood samples of rat were harvested for biochemical tests, as well as the liver and pancreas were isolated for histological examination and biochemical assays. The findings showed that puerarin-administered rats resulted in reduced glucose tolerance, blood insulin level, sero-enzymes of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and increased plasma level of high-density lipoprotein cholesterol (HDL-C) and reduced low-density lipoprotein cholesterol (LDL-C) content in serum. Further, the intrahepatic collagen deposits were lessened and positive cell of alpha smooth muscle actin (α-SMA) was lessened in puerarin treatment, while the pro-apoptotic cell numbers of Caspase 3, Bax in pancreatic islets were reduced dose-dependently. Moreover, the mRNA expressions of toll-like receptor 4 (TLR4) and tumor necrosis factor (TNF-α) in liver and pancreas were downregulated. In addition, TNF-α protein in the CCl4-lesioned liver and pancreas was reduced. Our findings demonstrate that puerarin contributes to attenuating the metabolic dysfunctions of CCl4-damaged liver and pancreas, in which the possible mechanisms may be linked to inhibition of inflammatory stress and normalization of metabolic homoeostasis in the liver and pancreas.

  18. Chemically-induced alteration of UDP-glucuronic acid concentration in rat liver.

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    Watkins, J B; Klaassen, C D

    1983-01-01

    Since many xenobiotics alter hepatic UDP-glucuronosyltransferase activity, their effect on UDPGA concentration was determined. Rats were pretreated with: 1) microsomal enzyme inducers (7,8-benzoflavone, benzo(a)pyrene, butylated hydroxyanisole, isosafrole, 3-methylcholanthrene, phenobarbital, pregnenolone-16 alpha-carbonitrile (PCN), 2,3,7,8-tetrachlorodibenzo-p-dioxin, trans-stilbene oxide); 2) inhibitors of microsomal enzymes (cobaltous chloride, piperonyl butoxide, SKF 525-A, borneol, galactosamine); 3) hepatotoxins (allyl alcohol, aflatoxin B1, alpha-naphthylisothiocyanate, bromobenzene, cadmium chloride, carbon tetrachloride, 1,1-dichloroethylene), and 4) commonly used anesthetics (pentobarbital, urethane, diethyl ether, halothane, enflurane, methoxyflurane). Rats were decapitated before removal of the liver. All inducers except PCN and isosafrole increased UDPGA 36-85% above control. Mixed-function oxidase inhibitors had no effect whereas borneol and galactosamine reduced UDPGA 85-90%. Aflatoxin B1 and cadmium produced decreases of 59 and 25%, respectively. Hepatic UDPGA content was diminished 70-95% after exposure to the inhalation anesthetics, whereas the other anesthetics reduced UDPGA about 25%. Thus, numerous xenobiotics alter the concentration of UDPGA in rat liver, which may influence the rate of glucoronidation.

  19. Dietary administration of inositol and/or inositol-6-phosphate prevents chemically-induced rat hepatocarcinogenesis.

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    Lee, Hae-Jeung; Lee, Sang-Ah; Choi, Haymie

    2005-01-01

    Chemoprevention is considered a rational strategy for dietary approaches to prevention of cancer. Multiple lines of evidence suggest that many of our dietary principles are able to intervene in the multistage carcinogenesis process and phytic acid (inositol hexaphosphate, IP6), a phytochemical present in a variety of plant species, has been shown to prevent various cancers, including those of the mammary gland, colon and liver. However, the mechanism of chemoprevention by IP6 has not been fully elucidated. In the present study, we examined the effects of inositol and/or IP6 supplementation on rat hepatocarcinogenesis initiated by diethylnitrosamine (DEN) and promoted by partial hepatectomy (PH). Supplementation with either inositol or IP6, or their combination, starting one week prior to administration of DEN, resulted in a significant decrease in both the area and the number of placental glutathione S-transferase positive (GST-P+) foci, a preneoplastic marker for DEN-initiated hepatocarcinogenesis. The administration of inositol and/or IP6 in drinking water caused marked enhancement in the glutathione S-transferase (GST) activity. In addition, the production of thiobarbituric acid reactive substances and the catalase activity were significantly reduced in rats supplemented with inositol and /or IP6. Based on these findings, it is likely that the chemopreventive effects of inositol and/or IP6 on rat hepatocarcinogenesis initiated by DEN and promoted by PH are associated with induction of GST activity and suppression of lipid peroxidation.

  20. Efficacy of Mesenchymal Stem Cells in Suppression of Hepatocarcinorigenesis in Rats: Possible Role of Wnt Signaling

    LENUS (Irish Health Repository)

    Abdel Aziz, Mohamed T

    2011-05-05

    Abstract Background The present study was conducted to evaluate the tumor suppressive effects of bone marrow derived mesenchymal stem cells (MSCs) in an experimental hepatocellular carcinoma (HCC) model in rats and to investigate the possible role of Wnt signaling in hepato-carcinogenesis. Methods Ninety rats were included in the study and were divided equally into: Control group, rats which received MSCs only, rats which received MSCs vehicle only, HCC group induced by diethylnitroseamine (DENA) and CCl 4 , rats which received MSCs after HCC induction, rats which received MSCs before HCC induction. Histopathological examination and gene expression of Wnt signaling target genes by real time, reverse transcription-polymerase chain reaction (RT-PCR) in rat liver tissue, in addition to serum levels of ALT, AST and alpha fetoprotein were performed in all groups. Results Histopathological examination of liver tissue from animals which received DENA-CCl4 only, revealed the presence of anaplastic carcinoma cells and macro-regenerative nodules type II with foci of large and small cell dysplasia. Administration of MSCs into rats after induction of experimental HCC improved the histopathological picture which showed minimal liver cell damage, reversible changes, areas of cell drop out filled with stem cells. Gene expression in rat liver tissue demonstrated that MSCs downregulated β-catenin, proliferating cell nuclear antigen (PCNA), cyclin D and survivin genes expression in liver tissues after HCC induction. Amelioration of the liver status after administration of MSCs has been inferred by the significant decrease of ALT, AST and Alpha fetoprotein serum levels. Administration of MSCs before HCC induction did not show any tumor suppressive or protective effect. Conclusions Administration of MSCs in chemically induced HCC has tumor suppressive effects as evidenced by down regulation of Wnt signaling target genes concerned with antiapoptosis, mitogenesis, cell proliferation

  1. Efficacy of Mesenchymal Stem Cells in Suppression of Hepatocarcinorigenesis in Rats: Possible Role of Wnt Signaling

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    Sabry Dina

    2011-05-01

    Full Text Available Abstract Background The present study was conducted to evaluate the tumor suppressive effects of bone marrow derived mesenchymal stem cells (MSCs in an experimental hepatocellular carcinoma (HCC model in rats and to investigate the possible role of Wnt signaling in hepato-carcinogenesis. Methods Ninety rats were included in the study and were divided equally into: Control group, rats which received MSCs only, rats which received MSCs vehicle only, HCC group induced by diethylnitroseamine (DENA and CCl4, rats which received MSCs after HCC induction, rats which received MSCs before HCC induction. Histopathological examination and gene expression of Wnt signaling target genes by real time, reverse transcription-polymerase chain reaction (RT-PCR in rat liver tissue, in addition to serum levels of ALT, AST and alpha fetoprotein were performed in all groups. Results Histopathological examination of liver tissue from animals which received DENA-CCl4 only, revealed the presence of anaplastic carcinoma cells and macro-regenerative nodules type II with foci of large and small cell dysplasia. Administration of MSCs into rats after induction of experimental HCC improved the histopathological picture which showed minimal liver cell damage, reversible changes, areas of cell drop out filled with stem cells. Gene expression in rat liver tissue demonstrated that MSCs downregulated β-catenin, proliferating cell nuclear antigen (PCNA, cyclin D and survivin genes expression in liver tissues after HCC induction. Amelioration of the liver status after administration of MSCs has been inferred by the significant decrease of ALT, AST and Alpha fetoprotein serum levels. Administration of MSCs before HCC induction did not show any tumor suppressive or protective effect. Conclusions Administration of MSCs in chemically induced HCC has tumor suppressive effects as evidenced by down regulation of Wnt signaling target genes concerned with antiapoptosis, mitogenesis, cell

  2. Resveratrol enhances the chemopreventive effect of celecoxib in chemically induced breast cancer in rats.

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    Kisková, Terézia; Jendželovský, Rastislav; Rentsen, Erdenetsetsek; Maier-Salamon, Alexandra; Kokošová, Natália; Papčová, Zuzana; Mikeš, Jaromír; Orendáš, Peter; Bojková, Bianka; Kubatka, Peter; Svoboda, Martin; Kajo, Karol; Fedoročko, Peter; Jäger, Walter; Ekmekcioglu, Cem; Kassayová, Monika; Thalhammer, Theresia

    2014-11-01

    Resveratrol and celecoxib were used as chemopreventive agents in animal models of carcinogenesis, and exert antiproliferative and proapoptotic effects on cancer cells. Therefore, the aim of this study was to evaluate whether combining resveratrol with celecoxib may exert more potent anticarcinogenic effects than the single agents. Mammary carcinogenesis was initiated in 70 female Sprague-Dawley rats with N-methyl-N-nitrosourea (NMU). The chemoprevention with resveratrol, celecoxib, and their combination started 2 weeks before the first carcinogen dose and lasted until the end of the experiment. Tumor incidence and frequency, latency period, tumor volume, the expression of cyclooxygenase 2 (COX2) and growth differentiation factor 15 (GDF15), and also the formation of reactive oxygen species were analyzed using different methods. In addition, the levels of resveratrol and its metabolites in blood and selected tumor tissues were determined by high-performance liquid chromatography. Finally, the anticancer effects of the reagents were studied in the human breast cancer cell line MCF-7. Celecoxib as a single agent significantly decreased tumor frequency, prolonged tumor latency, and decreased the total number of malignant tumors compared with the NMU conditions. Tumor volume was nonsignificantly reduced (0.68±0.25 vs. 0.93±0.28 cm3). Importantly, the addition of resveratrol to celecoxib reduced tumor volume by 60% compared with celecoxib alone (from 0.68±0.25 to 0.27±0.07 cm3, Pcancer-preventive effects of this application. This study showed that in NMU-induced mammary cancer in rats, the combination of resveratrol and celecoxib led to a significant reduction in all tumor parameters. In addition, in terms of tumor volume, the combination was more efficient than celecoxib as a single agent.

  3. Antineoplastic potential of Bryophyllum pinnatum lam. on chemically induced hepatocarcinogenesis in rats

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    Muhammad Afzal

    2013-01-01

    Full Text Available Background: Bryophyllum pinnatum Lam. used in folk medicine in tropical Africa, tropical America, India, China and Australia contains a wide range of active compounds, well known for their haemostatic and wound-healing properties. Objective: The present study was designed to evaluate the effect of Bryophyllum pinnatum Lam. on N-diethylnitrosamine (DENA-induced hepatic injury in rats. Material and Methods: The aerial part of B. pinnatum aqueous and ethanolic extract was prepared in doses of 250 mg/kg and 500 mg/kg. Hepatic injury was induced by DENA. Acute toxicity was also carried out. Result: Treatment with different doses of ethanolic extract of B. Pinnatum (250 mg/kg, p.o. was not significantly able to treat the liver injury induced by DENA, but 500 mg/kg dose of ethanolic extract of B. Pinnatum protects the liver slightly. Treatment with different doses of aqueous extract of B. Pinnatum (250 and 500 mg/kg, p.o. significantly (PFNx01<0.05; PFNx08<0.01 and PFNx18<0.001 treated the liver injury induced by DENA. Conclusion: It may be inferred from the present study that the hepatoprotective activities of the aqueous extract of B. Pinnatum leaves in DENA-induced hepatotoxicity may involve its antioxidant or oxidative free radical scavenging activities by alleviating lipid peroxidation through scavenging of free radicals, or by enhancing the activity of antioxidants.

  4. Rat epidermal keratinocyte organotypic culture (ROC) as a model for chemically induced skin irritation testing.

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    Pappinen, Sari; Pasonen-Seppänen, Sanna; Suhonen, Marjukka; Tammi, Raija; Urtti, Arto

    2005-11-01

    The potential of rat epidermal keratinocyte (REK) organotypic culture (ROC) with proper stratum corneum barrier as a model for screening skin irritants was evaluated. The test chemicals were selected from ECETOC database (1995) and the observed in vitro irritation potential was compared to ECETOC in vivo primary irritation index (PII), to EU risk phrases, and to the harmonized OECD criteria. Chemicals were applied onto the stratum corneum surface of ROC for 30 min and samples were taken from the underlying medium at 4 and 8 h after exposure. Cell membrane integrity (determined by LDH assay) and pro-inflammatory effect (determined by IL-1alpha release) were verified at both time points and correlated to PII values. The best correlation (R(2) = 0.831) was seen with LDH leakage test. Based on obtained data, chemicals were classified according to criteria defined by EU and OECD. From 12 chemicals, only two were incorrectly classified according to OECD criteria when using LDH leakage and IL-1alpha release as irritation markers. At the end of experiment, chemical-treated ROC cultures were fixed and histological changes were assessed. Typical signs for irritation were lightly stained cytoplasm, condensed nuclei, cellular vacuolization, eosinophilic cytoplasms, and blebbing. These irritation effects of chemicals were graded visually into four classes (A-D). The extent of morphological perturbations of the cultures mostly correlated with PII. The present results indicate the validity of the ROC model in predicting skin irritation potential of chemicals and show that the use of set of irritation markers with different mechanistic responses gives more information on irritation than if only one marker was used.

  5. DNA Microarray technology reveals similar gene expression patterns in rats with vitamin A deficiency and chemically induced colitis

    NARCIS (Netherlands)

    Nur, T.; Peijnenburg, A.A.C.M.; Noteborn, H.P.J.M.; Baykus, H.; Reifen, R.

    2002-01-01

    Previous studies suggest that vitamin A deficiency may induce or intensify inflammatory changes in the rat gastrointestinal system. The present study was designed to compare the expression profiles of rat models of vitamin A deficiency and induced colitis. cDNA-microarray technology was used to dete

  6. Enhancement of mouse germ cell-associated genes expression by injection of human umbilical cord mesenchymal stem cells into the testis of chemical-induced azoospermic mice

    Institute of Scientific and Technical Information of China (English)

    RuiFeng Yang; TaiHua Liu; Kai Zhao; ChengLiang Xiong

    2014-01-01

    Various methods are currently under investigation to preserve fertility in males treated with high‑dose chemotherapy and radiation for malignant and nonmalignant disorders. Human umbilical cord mesenchymal stem cells(HUC‑MSCs), which possess potent immunosuppressive function and secrete various cytokines and growth factors, have the potential clinical applications. As a potential alternative, we investigate whether injection of HUC‑MSCs into the interstitial compartment of the testes to promote spermatogenic regeneration efifciently. HUC‑MSCs were isolated from different sources of umbilical cords and injected into the interstitial space of one testis from 10 busulfan‑treated mice(saline and HEK293cells injections were performed in a separate set of mice) and the other testis remained uninjected. Three weeks after MSCs injection, Relative quantitative reverse transcription polymerase chain reaction was used to identify the expression of 10 of germ cell associated, which are all related to meiosis, demonstrated higher levels of spermatogenic gene expression(2–8 fold) in HUC‑MSCs injected testes compared to the contralateral uninjected testes(ifve mice). Protein levels for germ cell‑speciifc genes,miwi, vasa and synaptonemal complex protein (Scp3)were also higher in MSC‑treated testes compared to injected controls 3weeks after treatment. However, no different expression was detected in saline water and HEK293cells injection control group. We have demonstrated HUC‑MSCs could affect mouse germ cell‑speciifc genes expression. The results also provide a possibility that the transplanted HUC‑MSCs may promote the recovery of spermatogenesis. This study provides further evidence for preclinical therapeutic effects of HUC‑MSCs, and explores a new approach to the treatment of azoospermia.

  7. Effects of artificial cordyceps sinensis on epithelial-mesenchymal transition in the podocytes of diabetic rats

    Institute of Scientific and Technical Information of China (English)

    蔡芸莹

    2013-01-01

    Objective To assess the effects of artificial cordyceps sinensis(Jin shuibao) on the numbers of podocytes and epithelial-mesenchymal transition in diabetic rats. Methods Diabetes was induced by intraperitoneal injection of low dose streptozocin.

  8. Characterization of chemically induced ovarian carcinomas in an ethanol-preferring rat model: influence of long-term melatonin treatment.

    Directory of Open Access Journals (Sweden)

    Luiz Gustavo A Chuffa

    Full Text Available Ovarian cancer is the fourth most common cause of cancer deaths among women, and chronic alcoholism may exert co-carcinogenic effects. Because melatonin (mel has oncostatic properties, we aimed to investigate and characterize the chemical induction of ovarian tumors in a model of ethanol-preferring rats and to verify the influence of mel treatment on the overall features of these tumors. After rats were selected to receive ethanol (EtOH, they were surgically injected with 100 µg of 7,12-dimethyl-benz[a]anthracene (DMBA plus sesame oil directly under the left ovarian bursa. At 260 days old, half of the animals received i.p. injections of 200 µg mel/100 g b.w. for 60 days. Four experimental groups were established: Group C, rats bearing ovarian carcinomas (OC; Group C+EtOH, rats voluntarily consuming 10% (v/v EtOH and bearing OC; Group C+M, rats bearing OC and receiving mel; and Group C+EtOH+M, rats with OC consuming EtOH and receiving mel. Estrous cycle and nutritional parameters were evaluated, and anatomopathological analyses of the ovarian tumors were conducted. The incidence of ovarian tumors was higher in EtOH drinking animals 120 days post-DMBA administration, and mel efficiently reduced the prevalence of some aggressive tumors. Although mel promoted high EtOH consumption, it was effective in synchronizing the estrous cycle and reducing ovarian tumor mass by 20%. While rats in the C group displayed cysts containing serous fluid, C+EtOH rats showed solid tumor masses. After mel treatment, the ovaries of these rats presented as soft and mobile tissues. EtOH consumption increased the incidence of serous papillary carcinomas and sarcomas but not clear cell carcinomas. In contrast, mel reduced the incidence of sarcomas, endometrioid carcinomas and cystic teratomas. Combination of DMBA with EtOH intake potentiated the incidence of OC with malignant histologic subtypes. We concluded that mel reduces ovarian masses and the incidence of

  9. Characterization of Chemically Induced Ovarian Carcinomas in an Ethanol-Preferring Rat Model: Influence of Long-Term Melatonin Treatment

    Science.gov (United States)

    Chuffa, Luiz Gustavo A.; Fioruci-Fontanelli, Beatriz A.; Mendes, Leonardo O.; Fávaro, Wagner J.; Pinheiro, Patricia Fernanda F.; Martinez, Marcelo; Martinez, Francisco Eduardo

    2013-01-01

    Ovarian cancer is the fourth most common cause of cancer deaths among women, and chronic alcoholism may exert co-carcinogenic effects. Because melatonin (mel) has oncostatic properties, we aimed to investigate and characterize the chemical induction of ovarian tumors in a model of ethanol-preferring rats and to verify the influence of mel treatment on the overall features of these tumors. After rats were selected to receive ethanol (EtOH), they were surgically injected with 100 µg of 7,12-dimethyl-benz[a]anthracene (DMBA) plus sesame oil directly under the left ovarian bursa. At 260 days old, half of the animals received i.p. injections of 200 µg mel/100 g b.w. for 60 days. Four experimental groups were established: Group C, rats bearing ovarian carcinomas (OC); Group C+EtOH, rats voluntarily consuming 10% (v/v) EtOH and bearing OC; Group C+M, rats bearing OC and receiving mel; and Group C+EtOH+M, rats with OC consuming EtOH and receiving mel. Estrous cycle and nutritional parameters were evaluated, and anatomopathological analyses of the ovarian tumors were conducted. The incidence of ovarian tumors was higher in EtOH drinking animals 120 days post-DMBA administration, and mel efficiently reduced the prevalence of some aggressive tumors. Although mel promoted high EtOH consumption, it was effective in synchronizing the estrous cycle and reducing ovarian tumor mass by 20%. While rats in the C group displayed cysts containing serous fluid, C+EtOH rats showed solid tumor masses. After mel treatment, the ovaries of these rats presented as soft and mobile tissues. EtOH consumption increased the incidence of serous papillary carcinomas and sarcomas but not clear cell carcinomas. In contrast, mel reduced the incidence of sarcomas, endometrioid carcinomas and cystic teratomas. Combination of DMBA with EtOH intake potentiated the incidence of OC with malignant histologic subtypes. We concluded that mel reduces ovarian masses and the incidence of adenocarcinomas in

  10. Graptopetalum paraguayense ameliorates chemical-induced rat hepatic fibrosis in vivo and inactivates stellate cells and Kupffer cells in vitro.

    Directory of Open Access Journals (Sweden)

    Li-Jen Su

    Full Text Available BACKGROUND: Graptopetalum paraguayense (GP is a folk herbal medicine with hepatoprotective effects that is used in Taiwan. The aim of this study was to evaluate the hepatoprotective and antifibrotic effects of GP on experimental hepatic fibrosis in both dimethylnitrosamine (DMN- and carbon tetrachloride (CCl(4-induced liver injury rats. METHODS: Hepatic fibrosis-induced rats were fed with the methanolic extract of GP (MGP by oral administration every day. Immunohistochemistry, biochemical assays, and Western blot analysis were performed. The effects of MGP on the expression of fibrotic markers and cytokines in the primary cultured hepatic stellate cells (HSCs and Kupffer cells, respectively, were evaluated. RESULTS: Oral administration of MGP significantly alleviated DMN- or CCl(4-induced liver inflammation and fibrosis. High levels of alanine transaminase, aspartate transaminase, bilirubin, prothrombin activity and mortality rates also decreased in rats treated with MGP. There were significantly decreased hydroxyproline levels in therapeutic rats compared with those of the liver-damaged rats. Collagen I and alpha smooth muscle actin (α-SMA expression were all reduced by incubation with MGP in primary cultured rat HSCs. Furthermore, MGP induced apoptotic cell death in activated HSCs. MGP also suppressed lipopolysaccharide-stimulated rat Kupffer cell activation by decreasing nitric oxide, tumor necrosis factor-α and interleukin-6 production, and increasing interleukin-10 expression. CONCLUSIONS: The results show that the administration of MGP attenuated toxin-induced hepatic damage and fibrosis in vivo and inhibited HSC and Kupffer cell activation in vitro, suggesting that MGP might be a promising complementary or alternative therapeutic agent for liver inflammation and fibrosis.

  11. Hydroferrate fluid, MRN-100, provides protection against chemical-induced gastric and esophageal cancer in Wistar rats.

    Science.gov (United States)

    Ghoneum, Mamdooh H; Badr El-Din, Nariman K; Abdel Fattah, Salma M; Pan, Deyu; Tolentino, Lucilene

    2015-01-01

    In the current study, we examined the protective effect of hydroferrate fluid MRN-100 against the carcinogen methylnitronitrosoguanidine (MNNG)-induced gastric and esophageal cancer in rats. MRN-100 is an iron-based compound composed of bivalent and trivalent ferrates. At 33 weeks post treatment with MNNG, rats were killed and examined for the histopathology of esophagus and stomach; liver, spleen, and total body weight; and antioxidant levels in the blood and stomach tissues. Results showed that 17/20 (85%) gastroesophageal tissues from carcinogen MNNG-treated rats developed dysplasia and cancer, as compared to 8/20 (40%) rats treated with MNNG plus MRN-100. In addition, MRN-100 exerted an antioxidant effect in both the blood and stomach tissues by increasing levels of GSH, antioxidant enzymes SOD, CAT, and GPx, and total antioxidant capacity (TAC) level. This was accompanied by a reduction in the total free-radical and malondialdehyde levels. Furthermore, MRN-100 protected against body and organ weight loss. Thus, MRN-100 exhibited significant cancer chemopreventive activity by protecting tissues against oxidative damage in rats, which may suggest its effectiveness as an adjuvant for the treatment of gastric/esophageal carcinoma.

  12. Characterization Of Chemically Induced Ovarian Carcinomas In An Ethanol-preferring Rat Model: Influence Of Long-term Melatonin Treatment

    OpenAIRE

    Luiz Gustavo A Chuffa; Fioruci-Fontanelli, Beatriz A; Mendes, Leonardo O; Fávaro, Wagner J; Pinheiro, Patricia Fernanda F.; Marcelo Martinez; Francisco Eduardo Martinez

    2013-01-01

    Ovarian cancer is the fourth most common cause of cancer deaths among women, and chronic alcoholism may exert cocarcinogenic effects. Because melatonin (mel) has oncostatic properties, we aimed to investigate and characterize the chemical induction of ovarian tumors in a model of ethanol-preferring rats and to verify the influence of mel treatment on the overall features of these tumors. After rats were selected to receive ethanol (EtOH), they were surgically injected with 100 μg of 7,12-dime...

  13. Gravity, a regulation factor in the differentiation of rat bone marrow mesenchymal stem cells

    Directory of Open Access Journals (Sweden)

    Wan Yu-Min

    2009-09-01

    Full Text Available Abstract Background Stem cell therapy has emerged as a potential therapeutic option for tissue engineering and regenerative medicine, but many issues remain to be resolved, such as the amount of seed cells, committed differentiation and the efficiency. Several previous studies have focused on the study of chemical inducement microenvironments. In the present study, we investigated the effects of gravity on the differentiation of bone marrow mesenchymal stem cells (BMSCs into force-sensitive or force-insensitive cells. Methods and results Rat BMSCs (rBMSCs were cultured under hypergravity or simulated microgravity (SMG conditions with or without inducement medium. The expression levels of the characteristic proteins were measured and analyzed using immunocytochemical, RT-PCR and Western-blot analyses. After treatment with 5-azacytidine and hypergravity, rBMSCs expressed more characteristic proteins of cardiomyocytes such as cTnT, GATA4 and β-MHC; however, fewer such proteins were seen with SMG. After treating rBMSCs with osteogenic inducer and hypergravity, there were marked increases in the expression levels of ColIA1, Cbfa1 and ALP. Reverse results were obtained with SMG. rBMSCs treated with adipogenic inducer and SMG expressed greater levels of PPARgamma. Greater levels of Cbfa1- or cTnT-positive cells were observed under hypergravity without inducer, as shown by FACS analysis. These results indicate that hypergravity induces differentiation of rBMSCs into force-sensitive cells (cardiomyocytes and osteoblasts, whereas SMG induces force-insensitive cells (adipocytes. Conclusion Taken together, we conclude that gravity is an important factor affecting the differentiation of rBMSCs; this provides a new avenue for mechanistic studies of stem cell differentiation and a new approach to obtain more committed differentiated or undifferentiated cells.

  14. A robust method for assessing chemically induced mutagenic effects in the oral cavity of transgenic Big Blue® rats.

    Science.gov (United States)

    Young, Robert R; Thompson, Chad M; Dinesdurage, Harshini R; Elbekai, Reem H; Suh, Mina; Rohr, Annette C; Proctor, Deborah M

    2015-08-01

    The Big Blue® (BB) in vivo mutation assay uses transgenic rodents to measure treatment-induced mutations in virtually any tissue. The BB assay can be conducted in rats or mice and is ideal for investigating tissue-specific mutagenic mode of action of tumor induction. Some tissues such as oral mucosa have not been thoroughly studied. Due to the small quantity and cartilaginous nature of oral cavity tissues, development of special prosection and DNA isolation methods was required to permit robust analysis of mutations in these tissues. Improved surgical methods permitted collection of adequate and reproducible quantities of tissue (∼45 mg gingiva/buccal and ∼30 mg gingiva/palate). Optimized DNA isolation methods included use of liquid nitrogen pulverization, homogenization, nuclei pelleting, digestion, and phenol/chloroform extraction, to yield sufficient quantities of DNA from these tissues. In preliminary optimization work, mutant frequency (MF) in tongue and gingiva was increased in rats exposed to the promutagen, benzo[a]pyrene, and the direct mutagen, N-ethyl-N-nitrosourea. The oral cavity carcinogen, 4-nitroquinoline-1-oxide (4-NQO; 10 ppm in drinking water; 28 days), was qualified as a positive control for mutagenesis in oral tissues since it caused significant increases in cII MFs in gingiva/palate (50.2-fold) and gingiva/buccal tissues (21.3-fold), but not in liver or bone marrow (0.9- and 1.4-fold, respectively). These results are consistent with the observation that 4-NQO primarily induces tumors in oral cavity. Results also demonstrate the utility of the BB rat mutation assay and optimized methods for investigation of oral cavity mutagenicity, and by extension, analysis of other small and cartilaginous tissues.

  15. Edible oils for liver protection: hepatoprotective potentiality of Moringa oleifera seed oil against chemical-induced hepatitis in rats.

    Science.gov (United States)

    Al-Said, Mansour S; Mothana, Ramzi A; Al-Yahya, Mohammed A; Al-Blowi, Ali S; Al-Sohaibani, Mohammed; Ahmed, Atallah F; Rafatullah, Syed

    2012-07-01

    In the present study, in vitro antioxidant, antioxidative stress and hepatoprotective activity of Moringa oleifera Lam. seed oil (Ben oil; BO) was evaluated against carbon tetrachloride (CCl(4) ) induced lipid peroxidation and hepatic damage in rats. The oil at 0.2 and 0.4 mL/rat was administered orally for 21 consecutive days. The substantially elevated serum enzymatic (GOT, GPT, ALP, GGT) and bilirubin levels were significantly restored towards normalization by the oil. There was a significant elevation in the level of malondialdehyde (MDA), non-protein sulfhydryl (NP-SH), and total protein (TP) contents in the liver tissue. The results obtained indicated that BO possesses potent hepatoprotective action against CCl(4) -induced hepatic damage by lowering liver marker enzymes, MDA concentration, and elevating NP-SH and TP levels in liver tissue. The biochemical observations were supplemented with histopathological examination of rat liver. The results of this study showed that treatment with Ben oil or silymarin (as a reference) appears to enhance the recovery from hepatic damage induced by CCl(4) . The pentobarbital induced narcolepsy prolongation in mice was retarded by the Ben oil. Acute toxicity test in mice showed no morbidity or mortality. In vitro DPPH radical scavenging and β-carotene-linolic acid assay tests of the BO exhibited a moderate antioxidant activity in both tests used. The possible mechanism(s) of the liver protective activity of Ben oil activity may be due to free radical scavenging potential caused by the presence of antioxidant component(s) in the oil. Consequently, BO can be used as a therapeutic regime in treatment of some hepatic disorders.

  16. Oxymatrine could promote mesenchymal stem cell therapy in hepatic fibrosis rats:an experimental research

    Institute of Scientific and Technical Information of China (English)

    柴宁莉

    2013-01-01

    Objective To investigate whether oxymatrine (OM) could promote mesenchymal stem cell (MSC) therapy in CCl4-induced hepatic fibrosis (HF) in rats and to initially explore its mechanisms.Methods Totally 50 male SD rats were randomly divided into five groups,i.e.,nor-

  17. The effect of marrow mesenchymal stem cell transplantation on pulmonary fibrosis in rats

    Institute of Scientific and Technical Information of China (English)

    黄坤

    2012-01-01

    Objective To study the possible mechanisms of marrow mesenchymal stem cells(MSC) in therapy of bleomycin(BLM)-induced pulmonary fibrosis in rats. Methods Fifty-four female Wistar rats were randomly divided into a control group,a BLM group and a MSC group. The control group receivel intratracheal normal

  18. Isolation andin vivo hepatoprotective activity ofMelothria heterophylla (Lour.) Cogn. against chemically induced liver injuries in rats

    Institute of Scientific and Technical Information of China (English)

    Arijit Mondal; Tapan Kumar Maity; Dilipkumar Pal; Santanu Sannigrahi; Jagadish Singh

    2011-01-01

    Objective:To investigate hepatoprotective activity of ethanol extract of Melothria heterophylla Lour Cogn.(EEMH) againstCCl4-induced hepatic damage in rats.Methods:β-sitosterol was isolated by column chromatography and characterized spectroscopically. Two different doses (200 and400mg/kg bw) ofEEMHwere administered orally in alternate days. The hepatoprotective activity was studied in liver by measuring biochemical parameters such as serum aspartate amino transferase (AST), alanine amino transferase(ALT), alkaline phosphatase(ALP), total protein and total bilirubin. Lipid peroxidation product and different antioxidant enzyme activities were assessed in liver homogenate.Results:EEMH reduced all biochemical parameters and lipid peroxidation, as well as it increased the antioxidant enzyme activities in comparison with silymarin. The protective effect of the extract on CCl4 induced damage was confirmed by histopathological examination of the liver.Conclusions: This result strongly supports the protective effect ofEEMH against acute liver injury, and may be attributed to its antioxidative activity.

  19. Effects of metformin, buformin, and phenformin on the post-initiation stage of chemically induced mammary carcinogenesis in the rat.

    Science.gov (United States)

    Zhu, Zongjian; Jiang, Weiqin; Thompson, Matthew D; Echeverria, Dimas; McGinley, John N; Thompson, Henry J

    2015-06-01

    Metformin is a widely prescribed drug for the treatment of type II diabetes. Although epidemiologic data have provided a strong rationale for investigating the potential of this biguanide for use in cancer prevention and control, uncertainty exists whether metformin should be expected to have an impact in nondiabetic patients. Furthermore, little attention has been given to the possibility that other biguanides may have anticancer activity. In this study, the effects of clinically relevant doses of metformin (9.3 mmol/kg diet), buformin (7.6 mmol/kg diet), and phenformin (5.0 mmol/kg diet) were compared with rats fed control diet (AIN93-G) during the post-initiation stage of 1-methyl-1-nitrosourea-induced (50 mg/kg body weight) mammary carcinogenesis (n = 30/group). Plasma, liver, skeletal muscle, visceral fat, mammary gland, and mammary carcinoma concentrations of the biguanides were determined. In comparison with the control group, buformin decreased cancer incidence, multiplicity, and burden, whereas metformin and phenformin had no statistically significant effect on the carcinogenic process relative to the control group. Buformin did not alter fasting plasma glucose or insulin. Within mammary carcinomas, evidence was obtained that buformin treatment perturbed signaling pathways related to energy sensing. However, further investigation is needed to determine the relative contributions of host systemic and cell autonomous mechanisms to the anticancer activity of biguanides such as buformin.

  20. Non-Structural protein 1 (NS1) gene of Canine Parvovirus-2 regresses chemically induced skin tumors in Wistar rats.

    Science.gov (United States)

    Santra, Lakshman; Rajmani, R S; Kumar, G V P P S Ravi; Saxena, Shikha; Dhara, Sujoy K; Kumar, Amit; Sahoo, Aditya Prasad; Singh, Lakshya Veer; Desai, G S; Chaturvedi, Uttara; Kumar, Sudesh; Tiwari, Ashok K

    2014-10-01

    The Non-Structural protein 1 of Canine Parvovirus-2 (CPV2.NS1) plays a major role in viral cytotoxicity and pathogenicity. CPV2.NS1 has been proven to cause apoptosis in HeLa cells in vitro in our laboratory. Here we report that CPV2.NS1 has no toxic side effects on healthy cells but regresses skin tumors in Wistar rats. Histopathological examination of tumor tissue from CPV2.NS1 treated group revealed infiltration of mononuclear and polymorphonuclear cells with increased extra cellular matrix, indicating signs of regression. Tumor regression was also evidenced by significant decrease in mitotic index, AgNOR count and PCNA index, and increase in TUNEL positive apoptotic cells in CPV2.NS1 treated group. Further, CPV2.NS1 induced anti-tumor immune response through significant increase in CD8(+) and NK cell population in CPV2.NS1 treated group. These findings suggest that CPV2.NS1 can be a possible therapeutic candidate as an alternative to chemotherapy for the treatment of cancer.

  1. Expression of Neural Markers by Undifferentiated Rat Mesenchymal Stem Cells

    Directory of Open Access Journals (Sweden)

    Dana Foudah

    2012-01-01

    Full Text Available The spontaneous expression of neural markers by mesenchymal stem cells (MSCs has been considered to be a demonstration of MSCs’ predisposition to differentiate towards neural lineages. In view of their application in cell therapy for neurodegenerative diseases, it is very important to deepen the knowledge about this distinctive biological property of MSCs. In this study, we evaluated the expression of neuronal and glial markers in undifferentiated rat MSCs (rMSCs at different culture passages (from early to late. rMSCs spontaneously expressed neural markers depending on culture passage, and they were coexpressed or not with the neural progenitor marker nestin. In contrast, the number of rMSCs expressing mesengenic differentiation markers was very low or even completely absent. Moreover, rMSCs at late culture passages were not senescent cells and maintained the MSC immunophenotype. However, their differentiation capabilities were altered. In conclusion, our results support the concept of MSCs as multidifferentiated cells and suggest the existence of immature and mature neurally fated rMSC subpopulations. A possible correlation between specific MSC subpopulations and specific neural lineages could optimize the use of MSCs in cell transplantation therapy for the treatment of neurological diseases.

  2. Mesenchymal stem cell conditioning promotes rat oligodendroglial cell maturation.

    Directory of Open Access Journals (Sweden)

    Janusz Joachim Jadasz

    Full Text Available Oligodendroglial progenitor/precursor cells (OPCs represent the main cellular source for the generation of new myelinating oligodendrocytes in the adult central nervous system (CNS. In demyelinating diseases such as multiple sclerosis (MS myelin repair activities based on recruitment, activation and differentiation of resident OPCs can be observed. However, the overall degree of successful remyelination is limited and the existence of an MS-derived anti-oligodendrogenic milieu prevents OPCs from contributing to myelin repair. It is therefore of considerable interest to understand oligodendroglial homeostasis and maturation processes in order to enable the development of remyelination therapies. Mesenchymal stem cells (MSC have been shown to exert positive immunomodulatory effects, reduce demyelination, increase neuroprotection and to promote adult neural stem cell differentiation towards the oligodendroglial lineage. We here addressed whether MSC secreted factors can boost the OPC's oligodendrogenic capacity in a myelin non-permissive environment. To this end, we analyzed cellular morphologies, expression and regulation of key factors involved in oligodendroglial fate and maturation of primary rat cells upon incubation with MSC-conditioned medium. This demonstrated that MSC-derived soluble factors promote and accelerate oligodendroglial differentiation, even under astrocytic endorsing conditions. Accelerated maturation resulted in elevated levels of myelin expression, reduced glial fibrillary acidic protein expression and was accompanied by downregulation of prominent inhibitory differentiation factors such as Id2 and Id4. We thus conclude that apart from their suggested application as potential anti-inflammatory and immunomodulatory MS treatment, these cells might also be exploited to support endogenous myelin repair activities.

  3. Use of FK506 and bone marrow mesenchymal stem cells for rat hind limb allografts

    Institute of Scientific and Technical Information of China (English)

    Youxin Song; Zhujun Wang; Zhixue Wang; Hong Zhang; Xiaohui Li; Bin Chen

    2012-01-01

    Dark Agouti rat donor hind limbs were orthotopically transplanted into Lewis rat recipients to verify the effects of bone marrow mesenchymal stem cells on neural regeneration and functional recovery of allotransplanted limbs in the microenvironment of immunotolerance. bone marrow mesenchymal stem cells were intramuscularly (gluteus maximus) injected with FK506 (tacrolimus) daily, and were transplanted to the injured nerves. Results indicated that the allograft group not receiving therapy showed severe rejection, with transplanted limbs detaching at 10 days after transplantation with complete necrosis. The number of myelinated axons and Schwann cells in the FK506 and FK506 + bone marrow mesenchymal stem cells groups were significantly increased. We observed a lesser degree of gastrocnemius muscle degeneration, and increased polymorphic fibers along with other pathological changes in the FK506 + bone marrow mesenchymal stem cells group. The FK506 + bone marrow mesenchymal stem cells group showed significantly better recovery than the autograft and FK506 groups. The results demonstrated that FK506 improved the immune microenvironment. FK506 combined with bone marrow mesenchymal stem cells significantly promoted sciatic nerve regeneration, and improved sensory recovery and motor function in hind limb allotransplant.

  4. Human bone marrow mesenchymal stem cell transplantation attenuates axonal injur y in stroke rats

    Institute of Scientific and Technical Information of China (English)

    Yi Xu; Shiwei Du; Xinguang Yu; Xiao Han; Jincai Hou; Hao Guo

    2014-01-01

    Previous studies have shown that transplantation of human bone marrow mesenchymal stem cells promotes neural functional recovery after stroke, but the neurorestorative mechanisms remain largely unknown. We hypothesized that functional recovery of myelinated axons may be one of underlying mechanisms. In this study, an ischemia/reperfusion rat model was established using the middle cerebral artery occlusion method. Rats were used to test the hypothesis that in-travenous transplantation of human bone marrow mesenchymal stem cells through the femoral vein could exert neuroprotective effects against cerebral ischemia via a mechanism associated with the ability to attenuate axonal injury. The results of behavioral tests, infarction volume analysis and immunohistochemistry showed that cerebral ischemia caused severe damage to the myelin sheath and axons. After rats were intravenously transplanted with human bone marrow mesenchymal stem cells, the levels of axon and myelin sheath-related proteins, including mi-crotubule-associated protein 2, myelin basic protein, and growth-associated protein 43, were elevated, infarct volume was decreased and neural function was improved in cerebral ischemic rats. These ifndings suggest that intravenously transplanted human bone marrow mesenchymal stem cells promote neural function. Possible mechanisms underlying these beneifcial effects in-clude resistance to demyelination after cerebral ischemia, prevention of axonal degeneration, and promotion of axonal regeneration.

  5. Photofrin-mediated photodynamic therapy of chemically-induced premalignant lesions and squamous cell carcinoma of the palatal mucosa in rats

    NARCIS (Netherlands)

    Nauta, JM; vanLeengoed, HLLM; Witjes, MJH; Nikkels, PGJ; Vermey, A; Roodenburg, JLN

    1997-01-01

    Photodynamic therapy (PDT), an experimental cancer therapy, was studied in an animal model of chemically-induced epithelial dysplasia and squamous cell carcinoma. PDT was performed 24 hours after i.v. injection of 2.5 mg/kg bw Photofrin, and using 100 J/cm(2) incident light at two activation wavelen

  6. Autologous adipose tissue-derived mesenchymal stem cells are involved in rat liver regeneration following repeat partial hepatectomy

    OpenAIRE

    Liu, Tao; MU, HONG; Shen, Zhongyang; SONG, ZHUOLUN; Chen, Xiaobo; Wang, Yuliang

    2016-01-01

    Adipose tissue-derived mesenchymal stem cells (ADSCs) have been considered to be attractive and readily available adult mesenchymal stem cells, and they are becoming increasingly popular for use in regenerative cell therapy, as they are readily accessible through minimally invasive techniques. The present study investigated whether autologous ADSC transplantation promoted liver regeneration following a repeat partial hepatectomy in rats. The rats were divided into three groups as follows: 70%...

  7. Glucocorticoids induce autophagy in rat bone marrow mesenchymal stem cells

    DEFF Research Database (Denmark)

    Wang, L.; Fan, J.; Lin, Y. S.;

    2015-01-01

    and their responses to diverse stimuli, however, the role of autophagy in glucocorticoidinduced damage to bone marrow mesenchymal stem cells (BMSCs) remains unclear. The current study confirmed that glucocorticoid administration impaired the proliferation of BMSCs. Transmission electron microscopy......Glucocorticoidinduced osteoporosis (GIOP) is a widespread clinical complication following glucocorticoid therapy. This irreversible damage to boneforming and resorbing cells is essential in the pathogenesis of osteoporosis. Autophagy is a physiological process involved in the regulation of cells...

  8. Tangzhiqing Granules Alleviate Podocyte Epithelial-Mesenchymal Transition in Kidney of Diabetic Rats

    Directory of Open Access Journals (Sweden)

    Haiyan Xu

    2017-01-01

    Full Text Available This study discussed the effect of Tangzhiqing granules on podocyte epithelial-mesenchymal transition in kidney of diabetic rats. The diabetic rats were divided randomly into five groups: DM group treated with vehicle, Tangzhiqing granules low-dose treatment group, Tangzhiqing granules middle-dose treatment group, and Tangzhiqing granules high-dose treatment group. Eight Wistar rats used as control group were given saline solution. The intervention was all intragastric administration for 8 weeks. At the end of the 8 weeks, biochemical parameters and kidney weight/body weight ratio were measured. The kidney tissues were observed under light microscope and transmission electron microscopy. To search for the underlying mechanism, we examined the epithelial-to-mesenchymal transition (EMT related molecular markers and TGF-β/smad signaling pathway key proteins expression. The results showed that Tangzhiqing granules relieved the structural damage and functional changes of diabetic kidneys. Kidney podocyte EMT related molecular markers nephrin and CD2AP expression were increased, when desmin and α-SMA levels were decreased by Tangzhiqing granules in diabetic rats. Further TGF-β/smad signaling pathway key proteins TGF-β1 and p-smad2/3 levels were decreased in diabetic rats after treatment with Tangzhiqing granules. These findings suggest that Tangzhiqing granules may protect the podocytes of diabetic nephropathy rats via alleviating podocyte EMT and likely activating TGFβ/smad signaling pathway.

  9. A Survey of Mesenchyme-related Tumors of the Rat Kidney in the National Toxicology Program Archives, with Particular Reference to Renal Mesenchymal Tumor.

    Science.gov (United States)

    Hard, Gordon C; Seely, John Curtis; Betz, Laura J

    2016-08-01

    In order to harmonize diagnostic terminology, confirm diagnostic criteria, and describe aspects of tumor biology characteristic of different tumor types, a total of 165 cases of mesenchyme-related tumors and nephroblastomas of the rat kidney were reexamined from the National Toxicology Program (NTP) Archives. This survey demonstrated that renal mesenchymal tumor (RMT) was the most common spontaneous nonepithelial tumor in the rat kidney, also occurring more frequently in the NTP studies than nephroblastoma. Renal sarcoma was a distinct but very rare tumor entity, representing a malignant, monomorphous population of densely crowded, fibroblast-like cells, in which, unlike RMT, preexisting tubules did not persist. Nephroblastoma was characterized by early death of affected animals, suggesting an embryonal origin for this tumor type. Male and female rats were equally disposed to developing RMT, but most of the cases of nephroblastoma occurred in female rats and liposarcoma occurred mostly in male rats. This survey confirmed discrete histopathological and biological differences between the mesenchyme-related renal tumor types and between RMT and nephroblastoma. Statistical analysis also demonstrated a lack of any relationship of these renal tumor types to test article administration in the NTP data bank.

  10. Therapeutic effect of bone marrow mesenchymal stem cells on cold stress induced changes in the hippocampus of rats

    Institute of Scientific and Technical Information of China (English)

    Saravana Kumar Sampath Kumar; Saraswathi Perumal; Vijayaraghavan Rajagopalan

    2014-01-01

    The present study aims to evaluate the effect of bone marrow mesenchymal stem cells on cold stress induced neuronal changes in hippocampal CA1 region of Wistar rats. Bone marrow mes-enchymal stem cells were isolated from a 6-week-old Wistar rat. Bone marrow from adult femora and tibia was collected and mesenchymal stem cells were cultured in minimal essential medium containing 10% heat-inactivated fetal bovine serum and were sub-cultured. Passage 3 cells were analyzed by lfow cytometry for positive expression of CD44 and CD90 and negative expression of CD45. Once CD44 and CD90 positive expression was achieved, the cells were cultured again to 90% conlfuence for later experiments. Twenty-four rats aged 8 weeks old were randomly and evenly divided into normal control, cold water swim stress (cold stress), cold stress + PBS (intra-venous infusion), and cold stress + bone marrow mesenchymal stem cells (1 × 106; intravenous infusion) groups. The total period of study was 60 days which included 1 month stress period followed by 1 month treatment. Behavioral functional test was performed during the entire study period. After treatment, rats were sacriifced for histological studies. Treatment with bone marrow mesenchymal stem cells signiifcantly increased the number of neuronal cells in hippocampal CA1 region. Adult bone marrow mesenchymal stem cells injected by intravenous administration show potential therapeutic effects in cognitive decline associated with stress-related lesions.

  11. Mycophenolate mofetil ameliorates diabetic nephropathy through epithelial mesenchymal transition in rats.

    Science.gov (United States)

    Xiao, Xiaoyan; Wang, Jie; Chang, Xiangdi; Zhen, Junhui; Zhou, Gengyin; Hu, Zhao

    2015-09-01

    Recent studies in animal models have revealed that mycophenolate mofetil (MMF) has certain protective effects against experimental diabetic nephropathy. The present study therefore aimed to investigate the hypothesis that diabetic nephropathy may be ameliorated by mycophenolate mofetil and benazepril treatment alone or in combination, and identify the potential underlying mechanisms in a rat model. Diabetes was induced in rats by a single intraperitoneal injection of streptozotocin. Rats were subsequently treated with benazepril, MMF or a combination of the two drugs, and blood glucose, normalized kidney weight, urine protein and serum creatinine were determined. The pathological changes in renal tissue were also observed. In addition, indices of epithelial mesenchymal transition, including α‑smooth muscle actin (α‑SMA) and transforming growth factor (TGF)‑β1 expression, were examined. Normalized kidney weight, urine protein and serum creatinine levels were significantly improved in the diabetic rats treated with benazepril or mycophenolate mofetil, compared with those of rats in the untreated diabetic group. Pathological changes in the kidney were detected concurrently with increasing kidney weight and urinary albumin excretion, with a similar trend in variation among groups. In addition, the expression of epithelial mesenchymal transition indices, including α‑SMA and TGF‑β1, in the renal tubule interstitium were significantly decreased in the benazepril‑ and MMF‑treated groups compared with those of the diabetic group. As expected, the aforementioned indices were markedly lower in the benazepril and MMF combined treatment group than those in the single medication groups. These data suggested that MMF may have a protective role in diabetic nephropathy, and that the underlying mechanism may be partially dependent upon the suppression of the epithelial mesenchymal transition. Furthermore, the combination of benazepril and MMF conferred enhanced

  12. Transplanted bone marrow mesenchymal stem cells improve memory in rat models of Alzheimer's disease.

    Science.gov (United States)

    Babaei, Parvin; Soltani Tehrani, Bahram; Alizadeh, Arsalan

    2012-01-01

    The present study aims to evaluate the effect of bone marrow mesenchymal stem cells (MSCs) grafts on cognition deficit in chemically and age-induced Alzheimer's models of rats. In the first experiments aged animals (30 months) were tested in Morris water maze (MWM) and divided into two groups: impaired memory and unimpaired memory. Impaired groups were divided into two groups and cannulated bilaterally at the CA1 of the hippocampus for delivery of mesenchymal stem cells (500 × 10(3)/μL) and PBS (phosphate buffer saline). In the second experiment, Ibotenic acid (Ibo) was injected bilaterally into the nucleus basalis magnocellularis (NBM) of young rats (3 months) and animals were tested in MWM. Then, animals with memory impairment received the following treatments: MSCs (500 × 10(3)/μL) and PBS. Two months after the treatments, cognitive recovery was assessed by MWM in relearning paradigm in both experiments. Results showed that MSCs treatment significantly increased learning ability and memory in both age- and Ibo-induced memory impairment. Adult bone marrow mesenchymal stem cells show promise in treating cognitive decline associated with aging and NBM lesions.

  13. Transplanted Bone Marrow Mesenchymal Stem Cells Improve Memory in Rat Models of Alzheimer's Disease

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    Parvin Babaei

    2012-01-01

    Full Text Available The present study aims to evaluate the effect of bone marrow mesenchymal stem cells (MSCs grafts on cognition deficit in chemically and age-induced Alzheimer's models of rats. In the first experiments aged animals (30 months were tested in Morris water maze (MWM and divided into two groups: impaired memory and unimpaired memory. Impaired groups were divided into two groups and cannulated bilaterally at the CA1 of the hippocampus for delivery of mesenchymal stem cells (500×103/ and PBS (phosphate buffer saline. In the second experiment, Ibotenic acid (Ibo was injected bilaterally into the nucleus basalis magnocellularis (NBM of young rats (3 months and animals were tested in MWM. Then, animals with memory impairment received the following treatments: MSCs (500×103/ and PBS. Two months after the treatments, cognitive recovery was assessed by MWM in relearning paradigm in both experiments. Results showed that MSCs treatment significantly increased learning ability and memory in both age- and Ibo-induced memory impairment. Adult bone marrow mesenchymal stem cells show promise in treating cognitive decline associated with aging and NBM lesions.

  14. Mesenchymal stem cell infusion on skin wound healing of dexamethasone immunosuppressed wistar rats

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    Betânia Souza Monteiro

    Full Text Available ABSTRACT: To evaluate the therapeutic contribution of MSC intravenous infusion to surgical wound healing in dexamethasone-immunosuppressed rats, thirty-five rats were randomly divided into 2 groups: in the Control Group (CG, five rats received normal saline as 0.2ml subcutaneous (SC injections every 24 hours, for 30 consecutive days and, in the Dexamethasone Group (DG, 30 rats were given 0.2mL subcutaneous dexamethasone (0.1mg kg-1 every 24 hours, for 30 consecutive days. After 30 days, all rats underwent surgery to create an experimental skin wound. The 30 animals of the DG group were divided into two equal groups, which received different treatments: the dexamethasone group (DG received a single application of 0.5ml normal saline, via the intravenous route (IV, 48 hours after wound creation; and the Mesenchymal Stem Cells Dexamethasone group (MSCDG received MSC transplantation at a concentration of 9x106 cells in a single IV application, 48 hours after wound creation. The surgical wounds of CG rats closed on average 14.75 days after creation and DG rats had wounds closed within 22 days; whereas, the surgical wounds of MSCDG rats were closed in 14 days. MSC infusion in dexamethasone-immunosuppressed patients contributed positively to epithelial healing in less time.

  15. Pelvic Organ Distribution of Mesenchymal Stem Cells Injected Intravenously after Simulated Childbirth Injury in Female Rats

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    Michelle Cruz

    2012-01-01

    Full Text Available The local route of stem cell administration utilized presently in clinical trials for stress incontinence may not take full advantage of the capabilities of these cells. The goal of this study was to evaluate if intravenously injected mesenchymal stem cells (MSCs home to pelvic organs after simulated childbirth injury in a rat model. Female rats underwent either vaginal distension (VD or sham VD. All rats received 2 million GFP-labeled MSCs intravenously 1 hour after injury. Four or 10 days later pelvic organs and muscles were imaged for visualization of GFP-positive cells. Significantly more MSCs home to the urethra, vagina, rectum, and levator ani muscle 4 days after VD than after sham VD. MSCs were present 10 days after injection but GFP intensity had decreased. This study provides basic science evidence that intravenous administration of MSCs could provide an effective route for cell-based therapy to facilitate repair after injury and treat stress incontinence.

  16. Bone marrow mesenchymal stem cells protect against retinal ganglion cell loss in aged rats with glaucoma

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    Hu Y

    2013-10-01

    Full Text Available Ying Hu,1,2 Hai Bo Tan,1 Xin Mei Wang,3 Hua Rong,1 Hong Ping Cui,1 Hao Cui2 Departments of Ophthalmology, 1Shanghai East Hospital of Tongji University, Shanghai, 2First Affiliated Hospital, 3Fourth Affiliated Hospital, Harbin Medical University, Harbin, People's Republic of China Abstract: Glaucoma is a common eye disease in the aged population and has severe consequences. The present study examined the therapeutic effects of bone marrow mesenchymal stem cell (BMSC transplantation in preventing loss of visual function in aged rats with glaucoma caused by laser-induced ocular hypertension. We found that BMSCs promoted survival of retinal ganglion cells in the transplanted eye as compared with the control eye. Further, in swimming tests guided by visual cues, the rats with a BMSC transplant performed significantly better. We believe that BMSC transplantation therapy is effective in treating aged rats with glaucoma. Keywords: glaucoma, stem cell, transplantation, cell therapy, aging

  17. Differential effects of acute and repeated electrically and chemically induced seizures on ( sup 3 H)Nimodipine and ( sup 125 I)omega-conotoxin GVIA binding in rat brain

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    Gleiter, C.H.; Cain, C.J.; Weiss, S.R.; Post, R.M.; Marangos, P.J. (National Institute on Alcohol Abuse and Alcoholism, Bethesda, MD (USA))

    1989-07-01

    ({sup 3}H)Nimodipine and high-affinity ({sup 125}I)omega-conotoxin GVIA (CgTX) binding were investigated in membranes from rat cerebral cortex, cerebellum, and hippocampus after electrically and chemically induced seizures. Animals were decapitated 30 min after a single electroconvulsive shock (ECS) or lidocaine-induced seizure and 24 h after the last of 10 once-daily ECS or six once-daily lidocaine-induced seizures. After a single ECS, ({sup 3}H)nimodipine and ({sup 125}I)CgTX binding sites decreased in cerebral cortex (by 10% and 17%, respectively). A downregulation of ({sup 3}H)nimodipine binding sites in hippocampus occurred after single and repeated lidocaine-induced seizures (by 24% and 11%, respectively), whereas ({sup 125}I)CgTX binding remained unaltered. An earlier report on changes in ({sup 3}H)nitrendipine binding after chronic ECS in cortex and hippocampus was not confirmed.

  18. Effect of intravenous transplantation of bone marrow mesenchymal stem cells on neurotransmitters and synapsins in rats with spinal cord injury

    Institute of Scientific and Technical Information of China (English)

    Shaoqiang Chen; Bilian Wu; Jianhua Lin

    2012-01-01

    Bone marrow mesenchymal stem cells were isolated,purified and cultured in vitro by Percoll density gradient centrifugation combined with the cell adherence method.Passages 3-5 bone marrow mesenchymal stem cells were transplanted into rats with traumatic spinal cord injury via the caudal vein.Basso-Beattie-Bresnahan scores indicate that neurological function of experimental rats was significantly improved over transplantation time (1-5 weeks).Expressions of choline acetyltransferase,glutamic acid decarboxylase and synapsins in the damaged spinal cord of rats was significantly increased after transplantation,determined by immunofluorescence staining and laser confocal scanning microscopy.Bone marrow mesenchymal stem cells that had migrated into the damaged area of rats in the experimental group began to express choline acetyltransferase,glutamic acid decarboxylase and synapsins,3 weeks after transplantation.The Basso-Beattie-Bresnahan scores positively correlated with expression of choline acetyltransferase and synapsins.Experimental findings indicate that intravenously transplanted bone marrow mesenchymal stem cells traverse into the damaged spinal cord of rats,promote expression of choline acetyltransferase,glutamic acid decarboxylase and synapsins,and improve nerve function in rats with spinal cord injury.

  19. Polysaccharide hydrogel combined with mesenchymal stem cells promotes the healing of corneal alkali burn in rats.

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    Yifeng Ke

    Full Text Available Corneal chemical burns are common ophthalmic injuries that may result in permanent visual impairment. Although significant advances have been achieved on the treatment of such cases, the structural and functional restoration of a chemical burn-injured cornea remains challenging. The applications of polysaccharide hydrogel and subconjunctival injection of mesenchymal stem cells (MSCs have been reported to promote the healing of corneal wounds. In this study, polysaccharide was extracted from Hardy Orchid and mesenchymal stem cells (MSCs were derived from Sprague-Dawley rats. Supplementation of the polysaccharide significantly enhanced the migration rate of primarily cultured rat corneal epithelial cells. We examined the therapeutic effects of polysaccharide in conjunction with MSCs application on the healing of corneal alkali burns in rats. Compared with either treatment alone, the combination strategy resulted in significantly better recovery of corneal epithelium and reduction in inflammation, neovascularization and opacity of healed cornea. Polysaccharide and MSCs acted additively to increase the expression of anti-inflammatory cytokine (TGF-β, antiangiogenic cytokine (TSP-1 and decrease those promoting inflammation (TNF-α, chemotaxis (MIP-1α and MCP-1 and angiogenesis (VEGF and MMP-2. This study provided evidence that Hardy Orchid derived polysaccharide and MSCs are safe and effective treatments for corneal alkali burns and that their benefits are additive when used in combination. We concluded that combination therapy with polysaccharide and MSCs is a promising clinical treatment for corneal alkali burns and may be applicable for other types of corneal disorder.

  20. Efficacy of a mesenchymal stem cell loaded surgical mesh for tendon repair in rats

    OpenAIRE

    Schon, Lew C.; Gill, Nicholas; Thorpe, Margaret; Davis, Joel; Nadaud, Joshua; Kim, Jooyoung; Molligan, Jeremy; Zhang, Zijun

    2014-01-01

    Objectives The purpose of this study was to investigate the efficacy of a composite surgical mesh for delivery of mesenchymal stem cells (MSCs) in tendon repair. Methods The MSC-loaded mesh composed of a piece of conventional surgical mesh and a layer of scaffold, which supported MSC-embedded alginate gel. A 3-mm defect was surgically created at the Achilles tendon-gastrocnemius/soleus junction in 30 rats. The tendon defects were repaired with either 1) MSC-loaded mesh; or 2) surgical mesh on...

  1. Influence of age on rat bone-marrow mesenchymal stem cells potential.

    Science.gov (United States)

    Fafián-Labora, J; Fernández-Pernas, P; Fuentes, I; De Toro, J; Oreiro, N; Sangiao-Alvarellos, S; Mateos, J; Arufe, M C

    2015-11-19

    Mesenchymal stem cells promising role in cell-based therapies and tissue engineering appears to be limited due to a decline of their regenerative potential with increasing donor age. Six age groups from bone marrow mesenchymal stem cells of Wistar rats were studied (newborn, infant, young, pre-pubertal, pubertal and adult). Quantitative proteomic assay was performance by iTRAQ using an 8-plex iTRAQ labeling and the proteins differentially expressed were grouped in pluripotency, proliferative and metabolism processes. Proliferation makers, CD117 and Ki67 were measure by flow cytometry assay. Real time polymerase chain reaction analysis of pluripotency markers Rex1, Oct4, Sox2 and Nanog were done. Biological differentiation was realized using specific mediums for 14 days to induce osteogenesis, adipogenesis or chondrogenesis and immunostain analysis of differentiated cell resulting were done. Enzimoimmunoassay analysis of several enzymes as L-lactate dehydrogenase and glucose-6-phosphate isomerase were also done to validate iTRAQ data. Taking together these results indicate for the first time that mesenchymal stem cells have significant differences in their proliferative, pluripotency and metabolism profiles and those differences are age depending.

  2. Differentiation of rat marrow mesenchymal stem cells into pancreatic islet beta-cells

    Institute of Scientific and Technical Information of China (English)

    Li-Bo Chen; Xiao-Bing Jiang; Lian Yang

    2004-01-01

    AIM: To explore the possibility of marrow mesenchymal stem cells (MSC)in vitro differentiating into functional isletlike cells and to test the diabetes therapeutic potency of Islet-like cells.METHODS: Rat MSCs were isolated from Wistar rats and cultured. Passaged MSCs were induced to differentiate into islet-like cells under following conditions: pre-induction with L-DMEM including 10 mmol/L nicotinamide+1 mmol/L β-mercaptoethanol+200 mL/L fetal calf serum (FSC) for 24 h,followed by induction with serum free H-DMEM solution including 10 mmol/L nicotinamide+ 1 mmol/L,β-mercaptoethanol for 10 h. Differentiated cells were observed under inverse microscopy, insulin and nestin expressed in differentiated cells were detected with immunocytochemistry. Insulin excreted from differentiated cells was tested with radioimmunoassay. Rat diabetic models were made to test in vivo function of differentiated MSCs.RESULTS: Typical islet -like clustered cells were observed.Insulin mRNA and protein expressions were positive in differentiated cells, and nestin could be detected in predifferentiated cells. Insulin excreted from differentiated MSCs (446.93±102.28 IU/L) was much higher than that from pre-differentiated MSCs (2.45±0.81 IU/L (P<0.01).Injected differentiated MSCs cells could down-regulate glucose level in diabetic rats.CONCLUSION: Islet-like functional cells can be differentiated from marrow mesenchymal stem cells, which may be a new procedure for clinical diabetes stem -cell therapy, these cells can control blood glucose level in diabetic rats. MSCs may play an important role in diabetes therapy by islet differentiation and transplantation.

  3. Effect of dietary supplementation on the prognostic value of urinary and serum 8-isoprostaglandin F2α in chemically-induced mammary carcinogenesis in the rat

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    Białek Sławomir

    2011-03-01

    Full Text Available Abstract Backround The aim of the present study was to assess the effects of zinc or copper and polyphenolic compounds on the 8-isoprostaglandin F2α concentration in the serum and urine of rats with mammary cancer (adenocarcinoma induced with 7,12-dimethylbenz[a]antracene. The research focused on the kinetics of alterations in urinary 8-isoPGF2α at the early stage of carcinogenesis as well as the influence of dietary factors on the process. The impact of selected compounds on the intensity of DMBA - induced carcinogenesis was also assessed. Result and conclusions Administration of DMBA, a compound that inducers mammary tumors in experimental animals, increased the serum and urinary 8-isoPGF2α levels in study rats. In the rat model, diet supplementation with zinc, combined with selected polyphenolic compounds (resveratrol or genistein yielded a statistically significant decrease in the rat serum and urinary biomarker concentration with a simultaneously significant stimulation of carcinogenesis. The results indicate that there is an inverse correlation between the intensity of DMBA-induced carcinogenicity and the level of 8-isoPGF2α in urine and serum of rats.

  4. Chemically Induced Oncogenesis in the Peripheral Nervous System Is Suppressed in Congenic BDIX.BDIV-Mss1 and -Mss7 Rats

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    Bernd Koelsch

    2016-01-01

    Full Text Available Human malignant peripheral nerve sheath tumors (MPNSTs are highly aggressive soft-tissue sarcomas with a poor prognosis that arise either in the context of neurofibromatosis 1 or sporadically. Inbred BDIX and BDIV rat strains highly susceptible and resistant, respectively, to the development of ethylnitrosourea-induced MPNST enable us to identify, by using methods not applicable in humans, variant alleles involved in the pathways underlying individual MPNST risk. On the basis of a genome-wide association analysis using reciprocal intercrosses of BDIX and BDIV, BDIV alleles of two loci on chromosome 10, Mss1 and Mss7, were predicted to lower the risk of MPNST, the latter locus with a female bias. In this study we confirm the two nonoverlapping loci by exposing two congenic strains, BDIX.BDIV-Mss1 (Mss1 and BDIX.BDIV-Mss7 (Mss7, each carrying a BDIV genomic segment spanning the respective locus, to ethylnitrosourea. Compared with BDIX rats, the rate of MPNST is reduced 6.2-fold and 2.0-fold for Mss1 and Mss7 rats of both sexes, respectively. Although a moderate gain of survival time (30−50 days is seen in Mss1 rats of both sexes and Mss7 males, Mss7 females survive 134 days longer than BDIX females. BDIV alleles at Mss7 obviously cause a markedly increased intrastrain sex difference regarding survival time in Mss7 compared with BDIX rats. Fine mapping will lead to the identification of allelic variants modulating rat MPNST risk and subsequently to their human counterparts. This is of particular relevance, because so far neither gene nor anonymous sequence variants have been identified that influence the risk of human sporadic Schwann cell malignancy.

  5. Mesenchymal stem cells transplantation mildly ameliorates experimental diabetic nephropathy in rats

    Institute of Scientific and Technical Information of China (English)

    ZHOU Hong; TIAN Hao-ming; LONG Yang; ZHANG Xiang-xun; ZHONG Li; DENG Li; CHEN Xiao-he; LI Xiu-qun

    2009-01-01

    Background Diabetic nephropathy is a common complication of diabetes mellitus.This study aimed to explore whether mesenchymal stem cells(MSCs)transplantation could attenuate diabetic nephropathy in experimental diabetic rats.Methods Sprague-Dawley rats received a single intraperitoneal injection of streptozotocin(STZ)(60 mg/kg).Diabetic rats were randomized to four groups:diabetes control group(DC),ciclosporin A group(CsA),MSC group,and MSC+CsA group(MSCA).Bone marrow mesenchymal stern cells were cultured,identified and labeled by 5-bromo-2'-deoxyuridine(BrdU)in vitro.Then they were transplanted to diabetic rats via introcardiac infusion.Ciclosporin A was administered daily at 5 mg/kg.At 1,2,4,8 weeks after transplantation,random blood glucose,urine albumin/creatinine ratio(Alb/Cr),endogenous creatinine clearance rate and renal mass index were tested.Renal morphology and labeled cells were examined.Results Cultured MSCs expressed mesenchymal cell phenotype,and could be multidifferentiated to osteogenic and adipogenic cells.Labeled MSCs could be detected in the kidney of nephropathic rats,mainly in renal interstitium,but they did not propagate after engrafting in kidney.Over the course of the experiment,MSCA group showed a significant decrease in blood glucose compared with MSC group,CsA group and DC group(P<0.05,respectively).The Alb/Cr in MSCA group and MSC group were significantly lower than CsA group and DC group(P<0.05).And the Alb/Cr in MSCA group showed a significant decrease compared with MSC group(0.74 vs 0.84,P<0.05).There was a significant difference in renal mass index between the MSCA group and DC group(5.66 vs 6.37,P<0.05).No significant difference was found in creatinine clearance rate among 4 groups(P>0.05).Treatment with MSC+CsA significantly ameliorated the morphology of diabetic kidney.Conclusion MSC could mildly ameliorate diabetic nephropathy by decreasing blood glucose,Alb/Cr ratio and renal mass index.

  6. Decreased proliferation ability and differentiation potential of mesenchymal stem cells of osteoporosis rat

    Institute of Scientific and Technical Information of China (English)

    Qiang Wang; Bing Zhao; Chao Li; Jie-Sheng Rong; Shu-Qing Tao; Tian-Zun Tao

    2014-01-01

    Objective:To explore decreased proliferation ability and differentiation potential of mesenchymal stem cells(MSCs) of osteoporosis rat.Methods:MSCs were obtained from osteoporosis rat, and proliferation potency and impaired osteogenic differentiation potential were determined. Results:The result showed a significant downregulation ofMSCs pluripotency related gene(Oct 4) and osteogenic genes(BSP,OCN) expression inOVXMSCs compared withShamMSCs(P<0.05). Conclusions:These data suggest thatMSCs are aging in osteoporosis body, and autologous OVXMSCs transplantation is not appropriate to treat osteoporosis if necessary.There will be a possibility in establishing a new clinical application ofMSCs autologous transplantation to treat osteoporosis, ifOVXMSCs have stronger proliferation and differentiation.

  7. Does injection of metanephric mesenchymal cells improve renal function in rats?

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    Yu-qing Jiao

    2011-01-01

    Full Text Available Chronic kidney disease (CKD is a massive global health-care problem. Cell therapy offers a potential treatment for CKD. The aim of this study was to investigate whether the administration of a population of stem cells could be used to treat adriamycin (ADR-induced glomerulopathy in rats, a form of CKD. We intravenously transplanted metanephric mesenchymal cells (MMCs into rats treated with ADR. We also induced MMC differentiation in vitro using a medium derived from serum and homogenates of ADR-induced glomerulopathy rats. We detected the induction of an early epithelial phenotype (cytokeratin-18 expression and a proximal tubule phenotype (vitamin D receptor expression in vitro, and MMC-derived epithelial cells corresponding to the proximal tubule and glomeruli in vivo. Transplantation of MMCs after induction of glomerulopathy significantly increased the creatinine clearance rate (Ccr, a marker for glomerular filtration rate, but had no significant effect on other parameters (24-hour urinary protein excretion, serum albumin, total cholesterol. In addition, there was no significant difference in blood urea nitrogen or serum creatinine levels in rats with and without ADR administration. Our results indicate that MMCs might survive, engraft and differentiate into renal epithelia in vivo when transplanted into ADR-treated rats. However, further studies are needed to determine whether MMC transplantation improves renal function and causes renal repair in this model.

  8. Mesenchymal stem cells that located in the electromagnetic fields improves rat model of Parkinson's disease

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    Majid Jadidi

    2016-07-01

    Full Text Available Objective(s: The main characteristic of mesenchymal stem cells (MSCs is their ability to produce other cell types. Electromagnetic field (EMF stimulates differentiation of MSCs into other cells. In this study, we investigated whether EMF can effect on the differentiation of MSCs into dopaminergic (DA neurons. Materials and Methods: An EMF with a frequency of 50 Hz and two intensities of 40 and 400 µT 1hr/day was generated around the cells for a week. Afterwards, these cells were injected into the left ventricle of Parkinsonian rats. The rats survived for 2 weeks, and then sampling was performed. Results: The injected cells differentiated into DA neurons and sporadically settled in the substantia nigra pars compacta (SNpc. Transplanted rats exhibited significant partial correction apomorphine-induced rotational behavior compared to Parkinsonian rats (5.0±0.1 vs 7.57±0.08. Results demonstrated that endogenous serum and brain derived neurotrophic factor (BDNF were altered in all experimental groups. The greatest increase was in group of 400 µT EMF in comparison with Parkinsonian rats (398±15 vs. 312±11.79 pg ⁄ mg. Current study have shown that 6-Hydroxydopamine can cause severe loss of dopaminergic neurons (68±6.58, but injected MSCs that exposed to 40 and 400 µT EMF increased dopaminergic neurons in SNpc ( 108±2.33  & 126±3.89 (P

  9. Mesenchymal stem cells from rat olfactory bulbs can differentiate into cells with cardiomyocyte characteristics.

    Science.gov (United States)

    Huang, Yuahn-Sieh; Li, I-Hsun; Chueh, Sheau-Huei; Hueng, Dueng-Yuan; Tai, Ming-Cheng; Liang, Chang-Min; Lien, Shiu-Bii; Sytwu, Huey-Kang; Ma, Kuo-Hsing

    2015-12-01

    Mesenchymal stromal/stem cells (MSCs) are widely distributed in different tissues such as bone marrow, adipose tissues, peripheral blood, umbilical cord and amnionic fluid. Recently, MSC-like cells were also found to exist in rat olfactory bulb and are capable of inducing differentiation into mesenchymal lineages - osteocytes, chondrocytes and adipocytes. However, whether these cells can differentiate into myocardial cells is not known. In this study, we examined whether olfactory bulb-derived MSCs could differentiate into myocardial cells in vitro. Fibroblast-like cells isolated from the olfactory bulb of neonatal rats were grown under four conditions: no treatment; in the presence of growth factors (neuregulin-1, bFGF and forskolin); co-cultured with cardiomyocytes; and co-cultured with cardiomyocytes plus neuregulin-1, bFGF and forskolin. Cell differentiation into myocardial cells was monitored by RT-PCR, light microscopy immunofluorescence, western blot analysis and contractile response to pharmacological treatments. The isolated olfactory bulb-derived fibroblast-like cells expressed CD29, CD44, CD90, CD105, CD166 but not CD34 and CD45, consistent with the characteristics of MSCs. Long cylindical cells that spontaneously contracted were only observed following 7 days of co-culture of MSCs with rat cardiomyocytes plus neuregulin-1, bFGF and forskolin. RT-PCR and western blot analysis indicated that the cylindrical cells expressed myocardial markers, such as Nkx2.5, GATA4, sarcomeric α-actinin, cardiac troponin I, cardiac myosin heavy chain, atrial natriuretic peptide and connexin 43. They also contained sarcomeres and gap junction and were sensitive to pharmacological treatments (adrenal and cholinergic agonists and antagonists). These findings indicate that rat olfactory bulb-derived fibroblast-like cells with MSC characteristics can differentiate into myocardial-like cells.

  10. Human Umbilical Cord Mesenchymal Stem Cells Therapy in Cyclophosphamide-Induced Premature Ovarian Failure Rat Model

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    Dan Song

    2016-01-01

    Full Text Available Premature ovarian failure (POF is one of the most common causes of infertility in women. In our present study, we established cyclophosphamide- (CTX- induced POF rat model and elucidated its effect on ovarian function. We detected the serum estrogen, follicle stimulating hormone, and anti-Müllerian hormone of mice models by ELISA and evaluated their folliculogenesis by histopathology examination. Our study revealed that CTX administration could severely disturb hormone secretion and influence folliculogenesis in rat. This study also detected ovarian cells apoptosis by deoxy-UTP-digoxigenin nick end labeling (TUNEL and demonstrated marked ovarian cells apoptosis in rat models following CTX administration. In order to explore the potential of human umbilical cord mesenchymal stem cells (UCMSCs in POF treatment, the above indexes were used to evaluate ovarian function. We found that human UCMSCs transplantation recovered disturbed hormone secretion and folliculogenesis in POF rat, in addition to reduced ovarian cell apoptosis. We also tracked transplanted UCMSCs in ovaries by fluorescence in situ hybridization (FISH. The results manifested that the transplanted human UCMSCs could reside in ovarian tissues and could survive for a comparatively long time without obvious proliferation. Our present study provides new insights into the great clinical potential of human UCMSCs in POF treatment.

  11. Assessment of the effect of betaine on p16 and c-myc DNA methylation and mRNA expression in a chemical induced rat liver cancer model

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    Ling Wen-hua

    2009-07-01

    Full Text Available Abstract Background The development and progression of liver cancer may involve abnormal changes in DNA methylation, which lead to the activation of certain proto-oncogenes, such as c-myc, as well as the inactivation of certain tumor suppressors, such as p16. Betaine, as an active methyl-donor, maintains normal DNA methylation patterns. However, there are few investigations on the protective effect of betaine in hepatocarcinogenesis. Methods Four groups of rats were given diethylinitrosamine (DEN and fed with AIN-93G diets supplemented with 0, 10, 20 or 40 g betaine/kg (model, 1%, 2%, and 4% betaine, respectively, while the control group, received no DEN, fed with AIN-93G diet. Eight or 15 weeks later, the expression of p16 and c-myc mRNA was examined by Real-time PCR (Q-PCR. The DNA methylation status within the p16 and c-myc promoter was analyzed using methylation-specific PCR. Results Compared with the model group, numbers and areas of glutathione S-transferase placental form (GST-p-positive foci were decreased in the livers of the rats treated with betaine (P . Although the frequency of p16 promoter methylation in livers of the four DEN-fed groups appeared to increase, there is no difference among these groups after 8 or 15 weeks (P > 0.05. Betaine supplementation attenuated the down-regulation of p16 and inhibited the up-regulation of c-myc induced by DEN in a dose-dependent manner (P P . Finally, enhanced antioxidative capacity (T-AOC was observed in both the 2% and 4% betaine groups. Conclusion Our data suggest that betaine attenuates DEN-induced damage in rat liver and reverses DEN-induced changes in mRNA levels.

  12. Effect of zinc and polyphenols supplementation on antioxidative defense mechanisms and the frequency of microsatellite instability in chemically-induced mammary carcinogenesis in the rat.

    Science.gov (United States)

    Bobrowska-Korczak, Barbara; Skrajnowska, Dorota; Tokarz, Andrzej; Bialek, Slawomir; Jezierska, Ewelina

    2015-01-01

    The aim of the present study was to assess the effect of dietary supplementation (with zinc or zinc and polyphenolic compounds - resveratrol or genistein) on antioxidant enzymes (glutathione peroxidase - GPx, catalase - CAT and superoxide dismutase - SOD) and the frequency of microsatellite instability (MSI) in a widely used model of mammary carcinogenesis induced in the rat by treatment with 7,12-dimethyl-1,2-benz[a]anthracene (DMBA). The impact of selected compounds on the intensity of DMBA-induced carcinogenesis was also assessed. Sixty four Sprague-Dawley female rats were divided into study groups which, apart from the standard diet and DMBA, were treated with zinc, zinc and resveratrol or zinc and genistein via gavage for a period ranging from 40 days to 20 weeks of age. On the basis of the obtained results it can be said that synergistic reaction between Zn(II) and genistein causes a delay in cancer development as compared with the animals treated with DMBA but with no food supplementation. Supplementation with Zn(II) and polyphenolic compounds resulted in the occurrence of microsatellite instabilities in tumors. LOH (loss of heterozygosity) was found in tumor samples at microsatellite D1Mgh6 and D3Mgh9. DMBA treatment increased significantly the glutathione peroxidase activity whereas it had no effect on the SOD and CAT activities, as compared with control rats. Diet supplementation has an effect on the activity of selected antioxidant enzymes. Diet supplementation has an effect on the occurrence of microsatellite instabilities as well as on the intensity of the neoplastic process. The intensity of occurrence of microsatellite instabilities does not depend on the activity of selected antioxidant enzymes.

  13. Early combined treatment with sildenafil and adipose-derived mesenchymal stem cells preserves heart function in rat dilated cardiomyopathy

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    Fu Morgan

    2010-09-01

    Full Text Available Abstract Background We investigated whether early combined autologous adipose-derived mesenchymal stem cell (ADMSC and sildenafil therapy offers an additive benefit in preserving heart function in rat dilated cardiomyopathy (DCM. Methods Adult Lewis rats (n = 8 per group were divided into group 1 (normal control, group 2 (saline-treated DCM rats, group 3 [2.0 × 106 ADMSC implanted into left ventricular (LV myocardium of DCM rats], group 4 (DCM rats with sildenafil 30 mg/kg/day, orally, and group 5 (DCM rats with combined ADMSC-sildenafil. Treatment was started 1 week after DCM induction and the rats were sacrificed on day 90. Results The results showed that mitochondrial protein expressions of connexin43 and cytochrome-C were lowest in group 2, and lower in groups 3 and 4 than in group 5 (p Conclusion Early combined ADMSC/sildenafil is superior to either treatment alone in preserving LV function.

  14. Bone marrow-derived mesenchymal stem cells protect rats from endotoxin-induced acute lung injury

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    LIANG Zhi-xin; SUN Ji-ping; WANG Ping; TIAN Qing; YANG Zhen; CHEN Liang-an

    2011-01-01

    Background Acute lung injury (ALI) is a serious and common condition for which there are currently no specific strategies for treatment.Recent studies have suggested that bone marrow-derived multipotent mesenchymal stem cells (MSCs) may have therapeutic applications in multiple clinical disorders.We explored the biological effects of MSCs during endotoxin-induced ALl and the mechanisms involved.Methods MSCs were isolated from male rat bone marrow and the ALl model was induced by intravenous endotoxin injection.Female rats were sacrificed at 6 hours,24 hours,4 days,1 week and 3 weeks post-injection of MSCs or saline and the lung tissue,bronchoalveolar lavage fluid,and serum were harvested for analysis.We further evaluated the survival of the rats and examined the effects of endotoxin-induced injury on the interaction between alveolar macrophages (AMs) and MSCs in ex vivo.Results There was a significant decrease in numbers of neutrophils in bronchoalveolar lavage fluid (P <0.05),and myeloperoxidase activity in the lung (P<0.01),and of TNF-α and IL-1β in serum (P <0.05) in the MSC treated rats at 4 days.Furthermore,MSC treated rats exhibited improved survival,lower lung injury score,higher concentration of IL-10 in the serum and a reduced hydroxyproline content,but these differences were not statistically significant.Moreover,co-cultures of MSCs and AMs had significantly reduced levels of TNF-α,IL-1β and macrophage inflammatory protein (MIP)-1α and significantly increased levels of IL-10 (P<0.05) in the culture supernatants.Conclusions Treatment with intravenous injection of bone marrow-derived MSCs have beneficial effects on endotoxin-induced ALl in rats.The beneficial effect might be achieved through the engraftment of differentiated MSCs in the lungs and appears derive more from their capacity to secrete soluble factors that modulate immune responses.

  15. Intravenous transplantation of mesenchymal stem cells attenuates oleic acid induced acute lung injury in rats

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    XU Yu-lin; LIU Ying-long; WANG Qiang; LI Gang; L(U) Xiao-dong; KONG Bo

    2012-01-01

    Background Acute lung injury (ALI) and end-stage acute respiratory distress syndrome (ARDS) were among the most common causes of death in intensive care units.The activation of an inflammatory response and the damage of pulmonary epithelium and endotheliumwerethe hallmark of ALI/ARDS.Recent studies had demonstrated the importance of mesenchymal stem cells (MSCs) in maintaining the normal pulmonary endothelial and epithelial function as well as participating in modulating the inflammatory response and they are involved in epithelial and endothelial repair after injury.Here,our study demonstrates MSCs therapeutic potential in a rat model of ALI/ARDS.Methods Bone marrow derived MSCs were obtained from Sprague-Dawley (SD) rats and their differential potential was verified.ALl was induced in rats byoleic acid (OA),and MSCs were transplanted intravenously.The lung injury and the concentration of cytokines in plasma and lung tissue extracts were assessed at 8 hours,24 hours and 48 hours after OA-injection.Results The histological appearance and water content in rat lung tissue were significantly improved at different time points in rats treated with MSCs.The concentration of tumor necrosis factor-α and intercellular adhesion molecular-1 in rats plasma and lung tissue extracts were significantly inhibited after intravenous transplantation of MSCs,whereas interleukin-10 was significantly higher after MSCs transplantation at 8 hours,24 hours and 48 hours after OA-challenge.Conclusions Intravenous transplantation of MSCs could maintain the integrity of the pulmonary alveolar-capillary barrier and modulate the inflammatory response to attenuate the experimental ALI/ARDS.Transplantation of MSCs could be a novel cell-based therapeutic strategy for prevention and treatment of ALI/ARDS.

  16. Use of high pressure liquid chromatography to study chemically induced alterations in the pattern of benzo(a)pyrene metabolism. [Rat liver microsomes

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    Freudenthal, R.I.; Leber, A.P.; Emmerling, D.; Clarke, P.

    1975-11-01

    The metabolism of radiolabeled benzo(a)pyrene (BP) by control, 3-methylcholanthrene (3-MC) induced and 1,1,1-trichloropropene-2,3-oxide (TCPO)-inhibited rat liver microsomes was measured using fluorescence, radiometric, and high-pressure liquid chromatographic (HPLC) assays. Significant differences in the total measurable metabolism of BP by the three microsomal enzyme incubations resulted from the use of the three assay procedures. Appreciable differences in the concentration of the metabolite fractions after 3-MC induction and TCPO inhibition are clearly demonstrated. NMR analysis revealed that while the 3-hydroxy-BP fraction is greater than 90 percent pure, the 9-hydroxy fraction contains a number of metabolites having essentially identical retention times.

  17. Bone marrow-derived mesenchymal stem cells protect against experimental liver fibrosis in rats

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    Dong-Chang Zhao; Jun-Xia Lei; Rui Chen; Wei-Hua Yu; Xiu-Ming Zhang; Shu-Nong Li; Peng Xiang

    2005-01-01

    AIM: Recent reports have shown the capacity of mesenchymal stem cells (MSCs) to differentiate into hepatocytes in vitro and in vivo. MSCs administration could repair injured liver, lung, or heart through reducing inflammation, collagen deposition, and remodeling. These results provide a clue to treatment of liver fibrosis. The aim of this study was to investigate the effect of infusion of bone marrow (BM)-derived MSCs on the experimental liver fibrosis in rats.METHODS: MSCs isolated from BM in male Fischer 344 rats were infused to female Wistar rats induced with carbon tetrachloride (CCl4) or dimethylnitrosamine (DMN).There were two random groups on the 42nd d of CCl4:CCl4/MSCs, to infuse a dose of MSCs alone; CCl4/saline,to infuse the same volume of saline as control. There were another three random groups after exposure to DMN: DMN10/MSCs, to infuse the same dose of MSCs on d 10; DMN10/saline, to infuse the same volume of saline on d 10; DMN20/MSCs, to infuse the same dose of MSCson d 20. The morphological and behavioral changes ofrats were monitored everyday. After 4-6 wk of MSCs administration, all rats were killed and fibrosis index were assessed by histopathology and radioimmunoassay. Smooth muscle alpha-actin (alpha-SMA) of liver were tested by immunohistochemistry and quantified by IBAS 2.5 software. Male rats sex determination region on the Y chromosome (sry) gene were explored by PCR.RESULTS: Compared to controls, infusion of MSCsreduced the mortality rates of incidence in CCl4-induced model (10% vs 20%) and in DMN-induced model (2040% vs 90%).The amount of collagen deposition and alpha-SMA staining was about 40-50% lower in liver of rats with MSCs than that of rats without MSCs. The similar results were observed in fibrosis index. And the effect of the inhibition of fibrogenesis was greater in DMN10/MSCs than in DMN20/MSCs. The sry gene was positive in the liver of rats with MSCs treatment by PCR.CONCLUSION: MSCs treatment can protect against

  18. Effect of estrogen and antiestrogen therapy in rats bearing mesenchymal tumors.

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    Remedi, M M; Demarchi, M; Hliba, E; Depiante-Depaoli, M

    1999-01-01

    We applied both hormonal and antiestrogen treatment in female Wistar rats to analyze the estrogen dependence of the growth of sarcomas induced with 9,10-dimethyl-1,2-benzanthracene. Animals bearing tumors of 10 mm in diameter were divided at random into five groups and submitted to different treatments during 24 weeks. The treatment with ovariectomy and tamoxifen in tumor-bearing animals resulted in tumor growth suppression and prolonged survival by a protection against the lethal tumor. On the other hand, the estrogen treatment exerted an adverse effect showing a faster growth of the tumors and a great decrease in survival. In summary, the antiestrogen treatment can have an antitumor effect in mesenchymal tumors, possibly by modifying the immunological status of the host.

  19. The Three-Dimensional Collagen Scaffold Improves the Stemness of Rat Bone Marrow Mesenchymal Stem Cells

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    Sufang Han; Yannan Zhao; Zhifeng Xiao; Jin Han; Bing Chen; Lei Chen; Jianwu Dai

    2012-01-01

    Mesenchymal stem cells (MSCs) show the great promise for the treatment of a variety of diseases because of their self-renewal and multipotential abilities.MSCs are generally cultured on two-dimensional (2D) substrate in vitro.There are indications that they may simultaneously lose their stemness and multipotentiality as the result of prolonged 2D culture.In this study,we used three-dimensional (3D) collagen scaffolds as rat MSCs carrier and compared the properties of MSCs on 3D collagen scaffolds with monolayer cultured MSCs.The results demonstrated that collagen scaffolds were suitable for rat MSCs adherence and proliferation.More importantly,compared to MSCs under 2D culture,3D MSCs significantly maintained higher expression levels of stemness genes (Oct4,Sox2,Rex-1 and Nanog),yielded high frequencies of colony-forming units-fibroblastic (CFU-F) and showed enhanced osteogenic and adipogenic differentiation efficiency upon induction.Thus,3D collagen scaffolds may be beneficial for expanding rat MSCs while maintaining the stem cell properties in vitro.

  20. Improvement of Heart Failure by Human Amniotic Mesenchymal Stromal Cell Transplantation in Rats

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    Seyed Mohammad Taghi Razavi Tousi

    2016-11-01

    Full Text Available Background: Recently, stem cells have been considered for the treatment of heart diseases, but no marked improvement has been recorded. This is the first study to examine the functional and histological effects of the transplantation of human amniotic mesenchymal stromal cells (hAMSCs in rats with heart failure (HF.Methods: This study was conducted in the years 2014 and 2015. 35 male Wistar rats were randomly assigned into 5 equal experimental groups (7 rats each as 1- Control 2- Heart Failure (HF 3- Sham 4- Culture media 5- Stem Cell Transplantation (SCT. Heart failure was induced using 170 mg/kg/d of isoproterenol subcutaneously injection in 4 consecutive days. The failure confirmed by the rat cardiac echocardiography on day 28. In SCT group, 3×106 cells in 150 µl of culture media were transplanted to the myocardium. At the end, echocardiographic and hemodynamic parameters together with histological evaluation were done.Results: Echocardiography results showed that cardiac ejection fraction in HF group increased from 58/73 ± 9% to 81/25 ± 6/05% in SCT group (p value < 0.001. Fraction shortening in HF group was increased from 27/53 ± 8/58% into 45/55 ± 6/91% in SCT group (p value < 0.001. Furthermore, hAMSCs therapy significantly improved mean diastolic blood pressure, mean arterial pressure, left ventricular systolic pressure, rate pressure product, and left ventricular end-diastolic pressure compared to those in the HF group, with the values reaching the normal levels in the control group. A marked reduction in fibrosis tissue was also found in the SCT group (p value < 0.001 compared with the animals in the HF group.Conclusion: The transplantation of hAMSCs in rats with heart failure not only decreased the level of fibrosis but also conferred significant improvement in heart performance in terms of echocardiographic and hemodynamic parameters.

  1. Therapeutic effect of mesenchymal stem cells transplantation on pulmonary hypertension model rats

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    Ke-yan ZHAO

    2013-11-01

    Full Text Available Objective  To observe the therapeutic effect of mesenchymal stem cells (MSCs transplantation on monocrotaline (MCT-induced pulmonary hypertension in rats. Methods MSCs were isolated from Wistar rats and cultivated by bone marrow adherent culture. The third to fifth passages of MSCs were used for cell transplantation. Forty male Wistar rats were randomly divided into control group, MCT group, MCT/MSCs 5×105 group and MCT/MSCs 1×106 group (10 each. MCT was injected intraperitoneally (60mg/kg, and MSCs were transplanted into rats through external jugular vein. Right ventricular systolic pressure (RVSP of rats was determined, then the animals were sacrificed and the ventricular ratio (i.e. the mass ratio of right to left ventricle plus interventricular septum was calculated. The lung tissue was observed by light microscopy after HE staining, orcein staining and smooth muscle actin immunohistochemical staining. Results Four weeks after MSCs administration, the RVSP and ventricular ratio were 35.6±8.4mmHg and 0.357±0.092 in MCT/MSCs 1×106 group respectively, and significantly lower than those in MCT group (47.2±10.5mmHg and 0.445±0.094, respectively, P0.05. Pathological observation found the intima-media thickness of pulmonary arterioles was 19.2%±3.8% in MCT/MSCs 1×106 group, and significantly thinner than that in MCT group (26.4%±4.9%, P0.05. Conclusion  Intravenous MSCs administration could inhibit the MCT-induced pulmonary hypertension and 1×106 is the optimal number of MSCs for transplantation. DOI: 10.11855/j.issn.0577-7402.2013.10.010

  2. In vitro behaviors of rat mesenchymal stem cells on bacterial celluloses with different moduli

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    Taokaew, Siriporn [Department of Chemical Engineering, Faculty of Engineering, Chulalongkorn University, Bangkok 10330 (Thailand); Department of Chemical and Biomolecular Engineering, The University of Akron, Akron, OH 44325-3906 (United States); Phisalaphong, Muenduen [Department of Chemical Engineering, Faculty of Engineering, Chulalongkorn University, Bangkok 10330 (Thailand); Zhang Newby, Bi-min, E-mail: bimin@uakron.edu [Department of Chemical and Biomolecular Engineering, The University of Akron, Akron, OH 44325-3906 (United States)

    2014-05-01

    Compressive moduli of bacteria-synthesized cellulose (BC) were altered by two drying techniques: ambient-air drying and freeze drying. While no significant differences in dry weight were found, their cross-sectional structures and thickness varied greatly. Freeze dried BCs had loose cross-sectional structures and a thickness of ∼ 4.7 mm, whereas air dried BCs had more compacted cross-sectional structures and a thickness of ∼ 0.1 mm. The compressive moduli of the rehydrated freeze dried and rehydrated air dried BCs were measured to be 21.06 ± 0.22 kPa and 90.09 ± 21.07 kPa, respectively. When rat mesenchymal stem cells (rMSCs) were seeded on these BCs, they maintained a round morphology in the first 3 days of cultivation. More spread-out morphology and considerable proliferation on freeze dried BCs were observed in 7 days, but not on air-dried BCs. The cells were further grown for 3 weeks in the absence and presence of differentiation agents. Without using any differentiation agents, no detectable differentiation was noticed for rMSCs further cultivated on both types of BC. With differentiation inducing agents, chondrogenic differentiation, visualized by histological staining, was observed in some area of the rehydrated freeze dried BCs; while osteogenic differentiation was noticed on the stiffer rehydrated air dried BCs. - Graphical abstract: In the presence of induction agents, rat mesenchymal stem cells (rMSCs) preferentially differentiated into osteocytes on stiffer air dried BC films. - Highlights: • Bacterial cellulose (BC) sheets with different moduli generated by drying differently • Air-dried BC exhibited a modulus similar to that of bone. • Freeze-dried BC showed a modulus in the range of that of muscle. • Air-dried BC promoted the differentiation of rMSCs into osteocytes. • Freeze-dried BC promoted the differentiation of rMSCs into chondrocytes.

  3. Mesenchymal Stem Cells Inhibit Proteoglycan Degeneration in a Rat Model of Osteoarthritis

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    Zirak Javanmard

    2015-10-01

    Full Text Available Background Osteoarthritis (OA is a degenerative disease, which is characterized pathologically by degeneration of articular cartilage. One of the mechanisms of cartilage lesion in OA is decreased synthesis of cartilage matrix and amongst the variety of treatment methods for OA, cell base therapies has a crucial role. Objectives The purpose of this study was to understand the regeneration effect of mesenchymal stem cells as well as secretions of these cells after induction of early-stage Osteoarthritis (OA. Materials and Methods To achieve this aim articular cartilages of rat knees were cured by monosodium iodoacetate (1 mg/50 µL. Mesenchymal Stem Cells (MSCs were cultured and injected into the knee joint of rats after two weeks. The left knee was kept as the control group and injected with either sterill saline. The animals were sacrificed at two weeks after transplantation. The knee joints were harvested and safranin-o and toluidine-blue microscopic analysis were performed. In order to assess changes, the amount of control and experimental cartilage extracellular matrix structure (proteoglycan and cartilage fibrillation were determined by using histological approaches. Results The results revealed a decrease of proteoglycan content in the superficial and intermediate zones and presence of surface fibrillation. Therefore, OA induced matrix degeneration, and the reparative effects of MSCs were higher than cell secretions. Conclusions It can be concluded that due to some bioactive and nutrient factors, MSCs secretions could be used for recovery of OA as well as prevention of articular cartilage matrix degeneration, using a high dose of cell secretions.

  4. L-carnitine significantly decreased aging of rat adipose tissue-derived mesenchymal stem cells.

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    Mobarak, Halimeh; Fathi, Ezzatollah; Farahzadi, Raheleh; Zarghami, Nosratollah; Javanmardi, Sara

    2017-03-01

    Mesenchymal stem cells are undifferentiated cells that have the ability to divide continuously and tissue regeneration potential during the transplantation. Aging and loss of cell survival, is one of the main problems in cell therapy. Since the production of free radicals in the aging process is effective, the use of antioxidant compounds can help in scavenging free radicals and prevent the aging of cells. The aim of this study is evaluate the effects of L-carnitine (LC) on proliferation and aging of rat adipose tissue-derived mesenchymal stem cells (rADSC). rADSCs were isolated from inguinal region of 5 male Rattus rats. Oil red-O, alizarin red-S and toluidine blue staining were performed to evaluate the adipogenic, osteogenic and chondrogenic differentiation of rADSCs, respectively. Flow cytometric analysis was done for investigating the cell surface markers. The methyl thiazol tetrazolium (MTT) method was used to determine the cell proliferation of rADSCs following exposure to different concentrations of LC. rADSCs aging was evaluated by beta-galactosidase staining. The results showed significant proliferation of rADSCs 48 h after treatment with concentrations of 0.2 mM LC. In addition, in the presence of 0.2 mM LC, rADSCs appeared to be growing faster than control group and 0.2 mM LC supplementation could significantly decrease the population doubling time and aging of rADSCs. It seems that LC would be a good antioxidant to improve lifespan of rADSCs due to the decrease in aging.

  5. Human umbilical cord blood-derived mesenchymal stem cells promote regeneration of crush-injured rat sciatic nerves

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    Mi-Ae Sung; Jong-Ho Lee; Hun Jong Jung; Jung-Woo Lee; Jin-Yong Lee; Kang-Mi Pang; Sang Bae Yoo; Mohammad S. Alrashdan; Soung-Min Kim; Jeong Won Jahng

    2012-01-01

    Several studies have demonstrated that human umbilical cord blood-derived mesenchymal stem cells can promote neural regeneration following brain injury. However, the therapeutic effects of human umbilical cord blood-derived mesenchymal stem cells in guiding peripheral nerve regeneration remain poorly understood. This study was designed to investigate the effects of human umbilical cord blood-derived mesenchymal stem cells on neural regeneration using a rat sciatic nerve crush injury model. Human umbilical cord blood-derived mesenchymal stem cells (1 × 106) or a PBS control were injected into the crush-injured segment of the sciatic nerve. Four weeks after cell injection, brain-derived neurotrophic factor and tyrosine kinase receptor B mRNA expression at the lesion site was increased in comparison to control. Furthermore, sciatic function index, Fluoro Gold-labeled neuron counts and axon density were also significantly increased when compared with control. Our results indicate that human umbilical cord blood-derived mesenchymal stem cells promote the functional recovery of crush-injured sciatic nerves.

  6. Celecoxib in the histological grade of chemically induced mammary tumor in rats%塞莱昔布对化学诱导大鼠乳腺肿瘤病理分级影响的研究

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    钱跃清; 姚雨石; 于忠和; 杨光之; 战淑君

    2011-01-01

    OBJECTIVE: To evaluate the impact of celecoxib, a selective cyclooxygenase-2 inhibitor, on the histological grades of chemically induced mammary tumor and normal mammary tissue in rats, METHODS: Single dose of 7, 12 dimethlbenz anthracene, DMBA) was administered I. G. Into female SD rats to induce mammary tumors, during which various dose levels of celecoxib was dosed. The incidence, number and size of tumor were recorded, and mammary tumors and normal mammary tissues were analyzed via pathological observation. RESULTS: Compared with control group, 100 mg/kg and 150 mg/kg celecoxib treatment groups had significantly decreased incidence and number of tumor, while comparable tumor size. More importantly, celecoxib could significantly reduced histopathological grades of both mammary tumors (100 mg/kg, P = 0. 021; 150 mg/kg, P = 0. 036) and DMBA caused lesions in mammary lobule and alveolus (100 mg/kg, P = 0.004). CONCLUSIONS: Prophylactic administer of celecoxib can reduce pathological grade of chemically induced rat mammary tumor, as well as protecting normal mammary tissues. Its clinical application is worthy of further investigation.%目的:评价预防性应用选择性环氧化酶-2(COX-2)抑制剂塞莱昔布对化学诱导大鼠乳腺肿瘤及正常乳腺组织病理分级的影响.方法:雌性SD大鼠予二甲基苯并蒽(DMBA)1次性灌胃诱发乳腺肿瘤过程中,给予不同剂量塞莱昔布,观察肿瘤发生率、数量和体积,并与正常对照组进行病理学分析.结果:与对照组相比,塞莱昔布100和150 mg/kg两剂量组大鼠乳腺肿瘤发生率和肿瘤数量均显著低于模型对照组,但各组间肿瘤平均体积差异无统计学意义,P>0.05.塞莱昔布显著降低大鼠乳腺肿瘤的组织病理分级(100mg/kg,P=0.021;150 mg/kg,P=0.036),显著降低DMBA导致的乳腺小叶和腺泡的病变程度(100mg/kg,P=0.004).结论:预防性应用塞莱昔布具有降低乳腺肿瘤的病理分级,保护正常乳腺组织的作

  7. Nicorandil enhances the efficacy of mesenchymal stem cell therapy in isoproterenol-induced heart failure in rats.

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    Mohamed, Sarah S; Ahmed, Lamiaa A; Attia, Wael A; Khattab, Mahmoud M

    2015-12-01

    Stem cell transplantation has emerged as a promising technique for regenerative medicine in cardiovascular therapeutics. However, the results have been less than optimal. The aim of the present study was to investigate whether nicorandil could offer an additional benefit over bone marrow-derived mesenchymal stem cell therapy in isoproterenol-induced myocardial damage and its progression to heart failure in rats. Isoproterenol was injected subcutaneously for 2 consecutive days at doses of 85 and 170 mg/kg/day, respectively. Nicorandil (3 mg/kg/day) was then given orally with or without a single intravenous bone marrow-derived mesenchymal stem cell administration. Electrocardiography and echocardiography were recorded 2 weeks after the beginning of treatment. Rats were then sacrificed and the ventricle was isolated for estimation of tumor necrosis factor-alpha, vascular endothelial growth factor and transforming growth factor-beta. Moreover, protein expressions of caspase-3, connexin-43 as well as endothelial and inducible nitric oxide synthases were evaluated. Finally, histological studies of myocardial fibrosis and blood vessel density were performed and cryosections were done for estimation cell homing. Combined nicorandil/bone marrow-derived mesenchymal stem cell therapy provided an additional improvement compared to cell therapy alone toward reducing isoproterenol-induced cardiac hypertrophy, fibrosis and inflammation. Notably, combined therapy induced significant increase in angiogenesis and cell homing and prevented isoproterenol-induced changes in contractility and apoptotic markers. In conclusion, combined nicorandil/bone marrow-derived mesenchymal stem cell therapy was superior to cell therapy alone toward preventing isoproterenol-induced heart failure in rats through creation of a supportive environment for mesenchymal stem cells.

  8. Micro-Computed Tomography Detection of Gold Nanoparticle-Labelled Mesenchymal Stem Cells in the Rat Subretinal Layer

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    Pooi Ling Mok

    2017-02-01

    Full Text Available Mesenchymal stem cells are widely used in many pre-clinical and clinical settings. Despite advances in molecular technology; the migration and homing activities of these cells in in vivo systems are not well understood. Labelling mesenchymal stem cells with gold nanoparticles has no cytotoxic effect and may offer suitable indications for stem cell tracking. Here, we report a simple protocol to label mesenchymal stem cells using 80 nm gold nanoparticles. Once the cells and particles were incubated together for 24 h, the labelled products were injected into the rat subretinal layer. Micro-computed tomography was then conducted on the 15th and 30th day post-injection to track the movement of these cells, as visualized by an area of hyperdensity from the coronal section images of the rat head. In addition, we confirmed the cellular uptake of the gold nanoparticles by the mesenchymal stem cells using transmission electron microscopy. As opposed to other methods, the current protocol provides a simple, less labour-intensive and more efficient labelling mechanism for real-time cell tracking. Finally, we discuss the potential manipulations of gold nanoparticles in stem cells for cell replacement and cancer therapy in ocular disorders or diseases.

  9. Micro-Computed Tomography Detection of Gold Nanoparticle-Labelled Mesenchymal Stem Cells in the Rat Subretinal Layer

    Science.gov (United States)

    Mok, Pooi Ling; Leow, Sue Ngein; Koh, Avin Ee-Hwan; Mohd Nizam, Hairul Harun; Ding, Suet Lee Shirley; Luu, Chi; Ruhaslizan, Raduan; Wong, Hon Seng; Halim, Wan Haslina Wan Abdul; Ng, Min Hwei; Idrus, Ruszymah Binti Hj.; Chowdhury, Shiplu Roy; Bastion, Catherine Mae-Lynn; Subbiah, Suresh Kumar; Higuchi, Akon; Alarfaj, Abdullah A.; Then, Kong Yong

    2017-01-01

    Mesenchymal stem cells are widely used in many pre-clinical and clinical settings. Despite advances in molecular technology; the migration and homing activities of these cells in in vivo systems are not well understood. Labelling mesenchymal stem cells with gold nanoparticles has no cytotoxic effect and may offer suitable indications for stem cell tracking. Here, we report a simple protocol to label mesenchymal stem cells using 80 nm gold nanoparticles. Once the cells and particles were incubated together for 24 h, the labelled products were injected into the rat subretinal layer. Micro-computed tomography was then conducted on the 15th and 30th day post-injection to track the movement of these cells, as visualized by an area of hyperdensity from the coronal section images of the rat head. In addition, we confirmed the cellular uptake of the gold nanoparticles by the mesenchymal stem cells using transmission electron microscopy. As opposed to other methods, the current protocol provides a simple, less labour-intensive and more efficient labelling mechanism for real-time cell tracking. Finally, we discuss the potential manipulations of gold nanoparticles in stem cells for cell replacement and cancer therapy in ocular disorders or diseases. PMID:28208719

  10. Electrofusion of mesenchymal stem cells and islet cells for diabetes therapy: a rat model.

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    Goichi Yanai

    Full Text Available Islet transplantation is a minimally invasive treatment for severe diabetes. However, it often requires multiple donors to accomplish insulin-independence and the long-term results are not yet satisfying. Therefore, novel ways to overcome these problems have been explored. Isolated islets are fragile and susceptible to pro-apoptotic factors and poorly proliferative. In contrast, mesenchymal stem cells (MSCs are highly proliferative, anti-apoptotic and pluripotent to differentiate toward various cell types, promote angiogenesis and modulate inflammation, thereby studied as an enhancer of islet function and engraftment. Electrofusion is an efficient method of cell fusion and nuclear reprogramming occurs in hybrid cells between different cell types. Therefore, we hypothesized that electrofusion between MSC and islet cells may yield robust islet cells for diabetes therapy. We establish a method of electrofusion between dispersed islet cells and MSCs in rats. The fusion cells maintained glucose-responsive insulin release for 20 days in vitro. Renal subcapsular transplantation of fusion cells prepared from suboptimal islet mass (1,000 islets that did not correct hyperglycemia even if co-transplanted with MSCs, caused slow but consistent lowering of blood glucose with significant weight gain within the observation period in streptozotocin-induced diabetic rats. In the fusion cells between rat islet cells and mouse MSCs, RT-PCR showed new expression of both rat MSC-related genes and mouse β-cell-related genes, indicating bidirectional reprogramming of both β-cell and MSCs nuclei. Moreover, decreased caspase3 expression and new expression of Ki-67 in the islet cell nuclei suggested alleviated apoptosis and gain of proliferative capability, respectively. These results show that electrofusion between MSCs and islet cells yield special cells with β-cell function and robustness of MSCs and seems feasible for novel therapeutic strategy for diabetes

  11. Adipose tissue-derived mesenchymal stem cells repair germinal cells of seminiferous tubules of busulfan-induced azoospermic rats

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    Davood Mehrabani

    2015-01-01

    Full Text Available Context: Adipose tissue-derived mesenchymal stem cells (AT-MSCs are less invasive than bone marrow mesenchymal stem cells to obtain for cell therapy. Aims: The aims of this study were to evaluate the germinal cells characteristics and repairs in seminiferous tubules of busulfan-induced azoospermic rats after AT-MSCs transplantation. Settings and Design: Experimental case-control study. Materials and Methods: In the present experimental study, donors AT-MSCs were isolated from subcutaneous adipose tissue of two Sprague-Dawley rats. The recipients (n = 5 were received two doses of 10 mg/kg of busulfan with 21 days interval to stop endogenous spermatogenesis. After induction of azoospermia by busulfan, rats were injected with the AT-MSCs into the efferent duct of right testes. After 60 days, the right testes were injected AT-MSCs were compared to left azoospermic testes. Five untreated male rats served as negative control. Statistical Analysis Used: Stereological indices were analyzed by one-way ANOVA and LSD post-hoc test. The spermatogenesis index was compared using Mann-Whitney U test. Results: After stereological analyses, the seminiferous tubules treated with AT-MSCs had normal morphology. The untreated seminiferous tubules were empty. Spermatogenesis was observed in most cell-treated seminiferous tubules. Conclusions: The testis of busulfan-induced azoospermic rats accepted transplanted AT-MSCs. The transplanted AT-MSCs could induce spermatogenesis in seminiferous tubules of the rat.

  12. DAPI diffusion after intravitreal injection of mesenchymal stem cells in the injured retina of rats.

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    Castanheira, Paula; Torquetti, Leonardo Torquetti; Magalhãs, Débora Rodrigues Soares; Nehemy, Marcio B; Goes, Alfredo M

    2009-01-01

    To evaluate DAPI (4',6-diamidino-2-phenylindole) as a nuclear tracer of stem cell migration and incorporation it was observed the pattern of retinal integration and differentiation of mesenchymal stem cells (MSCs) injected into the vitreous cavity of rat eyes with retinal injury. For this purpose adult rat retinas were submitted to laser damage followed by transplantation of DAPI-labeled BM-MSCs grafts and double-labeled DAPI and quantum dot-labeled BM-MSCs. To assess a possible DAPI diffusion as well as the integration and differentiation of DAPI-labeled BM-MSCs in laser-injured retina, host retinas were evaluated 8 weeks after injury/transplantation. It was demonstrated that, 8 weeks after the transplant, most of the retinal cells in all neural retinal presented nuclear DAPI labeling, specifically in the outer nuclear layer (ONL), inner nuclear layer (INL), and ganglion cell layer (GCL). Meanwhile, at this point, most of the double-labeled BM-MSCs (DAPI and quantum dot) remained in the vitreous cavity and no retinal cells presented the quantum dot marker. Based on these evidences we concluded that DAPI diffused to adjacent retinal cells while the nanocrystals remained labeling only the transplanted BM-MSCs. Therefore, DAPI is not a useful marker for stem cells in vivo tracing experiments because the DAPI released from dying cells in moment of the transplant are taken up by host cells in the tissue.

  13. Infusion of Bone Marrow Mesenchymal Stem Cells Attenuates Experimental Severe Acute Pancreatitis in Rats

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    Huang, Dandan; Gao, Jun; Gong, Yanfang; Wu, Hongyu; Xu, Aifang

    2016-01-01

    Background & Aims. Severe acute pancreatitis (SAP) remains a high-mortality disease. Bone marrow (BM) mesenchymal stem cells (MSCs) have been demonstrated to have plasticity of transdifferentiation and to have immunomodulatory functions. In the present study, we assessed the roles of MSCs in SAP and the therapeutic effects of MSC on SAP after transplantation. Methods. A pancreatitis rat model was induced by the injection of taurocholic acid (TCA) into the pancreatic duct. After isolation and characterization of MSC from BM, MSC transplantation was conducted 24 hrs after SAP induction by tail vein injection. The survival rate was observed and MSCs were traced after transplantation. The expression of TNF-α and IL-1β mRNA in the transplantation group was also analyzed. Results. The survival rate of the transplantation group was significantly higher compared to the control group (p pancreas and BM 3 days after transplantation. The expression of TNF-α and IL-1β mRNA in the transplantation group was significantly lower than in the control group in both the pancreas and the lungs (p < 0.05). Conclusions. MSC transplantation could improve the prognosis of SAP rats. Engrafted MSCs have the capacity of homing, migration, and planting during the treatment of SAP. PMID:27721836

  14. Cysteine: A Novel Neural Inducer for Rat Bone Marrow Mesenchymal Stem Cells

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    Malek Soleimani Mehranjani

    2014-06-01

    Full Text Available Objective: Mesenchymal stem cells (MSCs can differentiate into various cell types. Since cysteine has structural similarities to neuronal inducers β-mercaptoethanol and glutathione, we examined its effect on neural induction of rat bone marrow MSCs. Materials and Methods: In this experimental study, cells were treated in a medium containing 1mM cysteine for 24 hours prior to treatment with neuron inducing medium containing 10 mM cysteine for 1, 2 and 3 hours. Cell viability and morphology were assessed by 3-(4,5-dimethylthiazol-2-Yl-2,5-diphenyltetrazolium bromide (MTT assay and, Hoechst, propidium iodide and acridine orange staining respectively. Expression of nestin and β-Tubulin III genes, as neural cell-specific markers, was studied reverse transcription polymerase chain reaction (RT-PCR. The data was statistically analyzed using One-Way ANOVA and Tukey’s test and p<0.05 was considered significant. Results: After 3 hours of treatment, neuron like morphology with a considerable expression of nestin and β-Tubulin III genes was apparent. The mean cell viability was not significantly different at 1, 2 and 3 hours following induction, compared with the control cells. Conclusion: Cysteine can induce neural features in rat bone marrow MSCs without reducing cell viability. Therefore, it can be considered as a safer alternative to toxic neural inducer agents such as β-mercaptoethanol.

  15. In vitro effect of direct current electrical stimulation on rat mesenchymal stem cells

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    Thottakkattumana Parameswaran, Vishnu; Barker, John Howard

    2017-01-01

    Background Electrical stimulation (ES) has been successfully used to treat bone defects clinically. Recently, both cellular and molecular approaches have demonstrated that ES can change cell behavior such as migration, proliferation and differentiation. Methods In the present study we exposed rat bone marrow- (BM-) and adipose tissue- (AT-) derived mesenchymal stem cells (MSCs) to direct current electrical stimulation (DC ES) and assessed temporal changes in osteogenic differentiation. We applied 100 mV/mm of DC ES for 1 h per day for three, seven and 14 days to cells cultivated in osteogenic differentiation medium and assessed viability and calcium deposition at the different time points. In addition, expression of osteogenic genes, Runx2, Osteopontin, and Col1A2 was assessed in BM- and AT-derived MSCs at the different time points. Results Results showed that ES changed osteogenic gene expression patterns in both BM- and AT-MSCs, and these changes differed between the two groups. In BM-MSCs, ES caused a significant increase in mRNA levels of Runx2, Osteopontin and Col1A2 at day 7, while in AT-MSCs, the increase in Runx2 and Osteopontin expression were observed after 14 days of ES. Discussion This study shows that rat bone marrow- and adipose tissue-derived stem cells react differently to electrical stimuli, an observation that could be important for application of electrical stimulation in tissue engineering.

  16. Tissue engineering of rat bladder using marrow-derived mesenchymal stem cells and bladder acellular matrix.

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    Daniel L Coutu

    Full Text Available Bladder replacement or augmentation is required in congenital malformations or following trauma or cancer. The current surgical solution involves enterocystoplasty but is associated with high complication rates. Strategies for bladder tissue engineering are thus actively sought to address this unmet clinical need. Because of the poor efficacy of synthetic polymers, the use of bladder acellular matrix (BAM has been proposed. Indeed when cellular components are removed from xenogenic or allogeneic bladders, the extracellular matrix scaffold thus obtained can be used alone or in combination with stem cells. In this study, we propose the use of BAM seeded with marrow-derived mesenchymal stem cells (MSCs for bladder tissue engineering. We optimized a protocol for decellularization of bladder tissue from different species including rat, rabbit and swine. We demonstrate the use of non-ionic detergents followed by nuclease digestion results in efficient decellularization while preserving the extracellular matrix. When MSCs were seeded on acellular matrix scaffold, they remained viable and proliferative while adopting a cellular phenotype consistent with their microenvironment. Upon transplantation in rats after partial cystectomy, MSC-seeded BAM proved superior to unseeded BAM with animals recovering nearly 100% normal bladder capacity for up to six months. Histological analyses also demonstrated increased muscle regeneration.

  17. Cultivation and optimized tracing of rat bone marrow mesenchymal stem cells

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    Xiu-hua HE

    2011-02-01

    Full Text Available Objective To investigate labelling and tracing methods of bone marrow mesenchymal stem cells(MSCs of rat,and to optimize the trace labelling technique.Methods Rat MSCs were isolated and cultured in vitro.The surface antigens(CD29,CD34,CD45,CD90 of MSCs were identified by flow cytometry,and MSCs were labelled with BrdU,DAPI and GFP,respectively.The labelling efficiency of BrdU was aseessed with immunocytochemistry,and that of DAPI and GFP were observed under fluorescence microscope.The advantages and disadvantages of the three tracer techniques were analyzed.Results Flow cytometry showed that MSCs expressed CD29 and CD90 but not CD34 or CD45.The three kinds of markers showed no significant toxicity to the cells.The optimal dosage and timing of BrdU labeling were respectively 10 μmol/L and 48 hours.And that of DAPI labeling were 1μg/ml and 12 hours.The infected MSCs with lentivirus-GFP at MOI(multiplicity of infection = 8 for 12h expressed GFP with high efficiency(above 90%.Conclusion Comparison with the three tracing methods for MSCs,transfection with GFP gene is a stable,reliable,safe tracing method,and they are important in tracing adult stem cells.

  18. Salvianolic acid B promotes survival of transplanted mesenchymal stem cells in spinal cord-injured rats

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    Xiao-bin BI; Yu-bin DENG; Dan-hui GAN; Ya-zhu WANG

    2008-01-01

    Aim: Stem cells hold great promise for brain and spinal cord injuries (SCI), but cell survival following transplantation to adult central nervous system has been poor. Salvianolic acid B (Sal B) has been shown to improve functional recovery in brain-injured rats. The present study was designed to determine whether Sal B could improve transplanted mesenchymal stem cell (MSC) survival in SCI rats. Methods: SCI rats were treated with Sal B. The Basso-Beatie-Bresnahan (BBB) scale was used to test the functional recovery. Sal B was used to protect MSC from being damaged by TNF-α in vitro. Bromodeoxyuridine-labeled MSC were transplanted into SCI rats with Sal B intraperitoneal injection, simul-taneously. MSC were examined, and the functional recovery of the SCI rats was tested. Results: Sal B treatment significantly reduced the lesion area from 0.26±0.05 mm2 to 0.15±0.03 mm2 (P<0.01) and remarkably raised the BBB scores on d 28, post-injury, from 7.3±0.9 to 10.5±1.3 (P<0.05), compared with the phosphate-buffered saline (PBS) control group. MSC were protected from the damage of TNF-α by Sal B. The number of surviving MSC in the MSC plus Sal B groups were 1143.3± 195.6 and 764.0±81.3 on d 7 and 28, post-transplantation, more than those in the MSC group, which was 569.3±72.3 and 237.0±61.3, respectively (P<0.05). Rats with MSC trans-planted and Sal B injected obtained higher BBB scores than those with MSC transplanted alone (P<0.05) and PBS (P<0.01). Conclusion: Sal B provides neuroprotection to SCI and promotes the survival of MSC in vitro and after cell transplantation to the injured spinal cord in vivo.

  19. Failure of intravenous or intracardiac delivery of mesenchymal stromal cells to improve outcomes after focal traumatic brain injury in the female rat.

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    L Christine Turtzo

    Full Text Available Mesenchymal stromal cells secrete a variety of anti-inflammatory factors and may provide a regenerative medicine option for the treatment of traumatic brain injury. The present study investigates the efficacy of multiple intravenous or intracardiac administrations of rat mesenchymal stromal cells or human mesenchymal stromal cells in female rats after controlled cortical impact by in vivo MRI, neurobehavior, and histopathology evaluation. Neither intravenous nor intracardiac administration of mesenchymal stromal cells derived from either rats or humans improved MRI measures of lesion volume or neurobehavioral outcome compared to saline treatment. Few mesenchymal stromal cells (<0.0005% of injected dose were found within 3 days of last dosage at the site of injury after either delivery route, with no mesenchymal stromal cells being detectable in brain at 30 or 56 days post-injury. These findings suggest that non-autologous mesenchymal stromal cells therapy via intravenous or intracardiac administration is not a promising treatment after focal contusion traumatic brain injury in this female rodent model.

  20. Do adipose tissue-derived mesenchymal stem cells ameliorate Parkinson's disease in rat model?

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    Ahmed, Hh; Salem, Am; Atta, Hm; Ghazy, Ma; Aglan, Ha

    2014-12-01

    Parkinson's disease (PD) is a common neurodegenerative disorder in middle-aged and elderly people. This study aimed to elucidate the role of mesenchymal stem cells (MSCs) in management of PD in ovariectomized rat model. MSCs were excised from adipose tissue of both the omentum and the inguinal fat pad of male rats, grown, and propagated in culture; then characterized morphologically; and by the detection of surface markers gene expression. In this study, 40 ovariectomized animals were classified into 5 groups; group 1 was ovariectomized control, groups 2 to 5 were subcutaneously administered with rotenone for 14 days after 1 month of ovariectomy for induction of PD. Group 2 was left untreated; groups 3, 4, and 5 were treated with Sinemet(®), Cerebrolysin(®), and a single dose of adipose tissue-derived MSCs (ADMSCs), respectively. Y-chromosome gene (sry) was assessed by polymerase chain reaction (PCR) in brain tissue of the female rats. Serum transforming growth factor β (TGF-β), monocyte chemoattractant protein 1 (MCP-1), and brain-derived neurotrophic factor (BDNF) levels were assayed using enzyme-linked immunosorbent assay technique. Brain dopamine level was assayed fluorometrically, while brain tyrosine hydroxylase (TH) gene expression was detected by semiquantitative real-time PCR. The PD group showed significant increase in serum TGF-β and MCP-1 levels associated with significant decrease in serum BDNF, brain dopamine, and brain TH gene expression levels. In contrast, all treatments produce significant decrease in serum TGF-β and MCP-1 levels in concomitant with significant increase in serum BDNF, brain dopamine, and brain TH gene expression levels. In conclusion, the observed improvements in the studied biomarkers due to ADMSCs infusion might be attributed to their immunomodulatory, anti-inflammatory, and neurotrophic effects.

  1. Radix Astragali-induced differentiation of rat bone marrow-derived mesenchymal stem cells

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    Xinsheng Wang; Haifeng Li; Ying Zhao; Xiaoli Zhang; Aihua Bo

    2009-01-01

    BACKGROUND: Chemical induction has been shown to be effective at promoting the differentiation of bone marrow-derived mesenchymal stem cells (MSCs). However, these inductors have cytotoxicity side effects that may damage cells over time. Traditional Chinese medicines avoid this disadvantage while still producing effective induction.OBJECTIVE: To investigate the influence of Radix Astragali (Huangqi) on the differentiation of MSCs.DESIGN, TIME AND SETTING: In vitro study of traditional Chinese medicine in neural stem cell differentiation. The experiment was performed at the Central Laboratory of Hebei North University between April and June 2007.MATERIALS: Radix Astragafi solution (lot No. 060105; license No. Z53021585) was purchased from Daii Pharmaceutical Co., Ltd., China; rabbit anti-rat nestin, rabbit anti-rat neuron-specific enolase (NSE), mouse anti-rat microtubule-associated protein 2, and rabbit anti-rat glial fibrillary acidic protein were purchased from Wuhan Boster, China.METHODS: Whole bone marrow was isolated from the femur and tibia of 6-week-old male Wistar rats and subcultured. The fourth passage of MSCs were harvested and induced by different concentrations (50, 100, 200, 400 g/L) of Radix Astragali.MAIN OUTCOME MEASURES: Hematoxylin-eosin staining was used to observe MSC morphology after 24 hours of induction. Immunocytochemistry was employed to observe the expression of NSE (specific neuronal marker), nestin (marker of neural stem cell), glial fibrillary acidic protein and microtubule-associated protein 2 (markers of astrocytes).RESULTS: Following Radix Astragafi treatment, changes occurred in cell morphology including: cell body pyknosis; thin and long processes formed in some cells, with growth corresponding to drug concentration and induction time; and the formation of network-like connections between some cells.With increasing drug concentration and induction time, nestin expression was upregulated, and the number of positive cells increased

  2. Plasmid-encapsulated polyethylene glycol-grafted polyethylenimine nanoparticles for gene delivery into rat mesenchymal stem cells

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    Chen X

    2011-04-01

    Full Text Available Xiao-Ai Chen1,5*, Li-Jun Zhang2*, Zhi-Jie He3, Wei-Wei Wang4, Bo Xu1, Qian Zhong1, Xin-Tao Shuai4, Li-Qun Yang4, Yu-Bin Deng11Department of Pathophysiology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China; 2Futian Affiliated Hospital, Guangdong Medical College, Shenzhen, China; 3Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China; 4Institute of Polymer Science, School of Chemistry and Chemical Engineering, BME Center, State Key Laboratory of Optoelectronic Materials and Technologies, Sun Yat-sen University, Guangzhou, China; 5Yunnan Cancer Hospital, The Third Affiliated Hospital, Kunming Medical College, Kunming, China*Both authors contributed equally to this workBackground: Mesenchymal stem cell transplantation is a promising method in regenerative medicine. Gene-modified mesenchymal stem cells possess superior characteristics of specific tissue differentiation, resistance to apoptosis, and directional migration. Viral vectors have the disadvantages of potential immunogenicity, carcinogenicity, and complicated synthetic procedures. Polyethylene glycol-grafted polyethylenimine (PEG-PEI holds promise in gene delivery because of easy preparation and potentially targeting modification.Methods: A PEG8k-PEI25k graft copolymer was synthesized. Agarose gel retardation assay and dynamic light scattering were used to determine the properties of the nanoparticles. MTT reduction, wound and healing, and differentiation assays were used to test the cytobiological characteristics of rat mesenchymal stem cells, fluorescence microscopy and flow cytometry were used to determine transfection efficiency, and atomic force microscopy was used to evaluate the interaction between PEG-PEI/plasmid nanoparticles and mesenchymal stem cells.Results: After incubation with the copolymer, the bionomics of mesenchymal stem cells showed no significant change. The mesenchymal stem cells still maintained high viability, resettled the

  3. A comparative study on the transplantation of different concentrations of human umbilical mesenchymal cells into diabetic rats

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    Jia-Hui; Kong; Dan; Zheng; Song; Chen; Hong-Tao; Duan; Yue-Xin; Wang; Meng; Dong; Jian; Song

    2015-01-01

    AIM: To observe the effects of intravitreal injections of different concentrations of human umbilical mesenchymal stem cells on retinopathy in rats with diabetes mellitus.METHODS: Healthy and adult male Sprague-Dawley(SD) rats were randomly assigned to a normal control group(group A), a diabetic retinopathy(DR) blank control group(group B), a high-concentration transplantation group(group C), a low-concentration transplantation group(group D) and a placebo transplantation group(group E). The expression of nerve growth factor(NGF)protein in the retinal layers was detected by immunohistochemical staining at 2, 4, 6 and 8wk.RESULTS: The expression of NGF was positive in group A and most positive in the retinal ganglion cell layer. In groups B and E, the expression of NGF was positive 2wk after transplantation and showed an increase in all layers. However, the level of expression had decreased in all layers at 4wk and was significantly reduced at 8wk. In groups C and D, the expression of NGF had increased at 2wk and continued to increase up to 8wk. The level of expression in group C was much higher than that in group D.CONCLUSION: DR can be improved by intravitreal injection of human umbilical mesenchymal stem cells.High concentrations of human umbilical mesenchymal stem cells confer a better protective effect on DR than low concentrations.

  4. Bone marrow mesenchymal stem cells overexpressing human basic fibroblast growth factor increase vasculogenesis in ischemic rats

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    Zhang, J.C. [Department of Vascular Surgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou (China); Zheng, G.F. [Department of Vascular Surgery, The People' s Hospital of Ganzhou, Ganzhou (China); Wu, L.; Ou Yang, L.Y.; Li, W.X. [Department of Vascular Surgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou (China)

    2014-08-08

    Administration or expression of growth factors, as well as implantation of autologous bone marrow cells, promote in vivo angiogenesis. This study investigated the angiogenic potential of combining both approaches through the allogenic transplantation of bone marrow-derived mesenchymal stem cells (MSCs) expressing human basic fibroblast growth factor (hbFGF). After establishing a hind limb ischemia model in Sprague Dawley rats, the animals were randomly divided into four treatment groups: MSCs expressing green fluorescent protein (GFP-MSC), MSCs expressing hbFGF (hbFGF-MSC), MSC controls, and phosphate-buffered saline (PBS) controls. After 2 weeks, MSC survival and differentiation, hbFGF and vascular endothelial growth factor (VEGF) expression, and microvessel density of ischemic muscles were determined. Stable hbFGF expression was observed in the hbFGF-MSC group after 2 weeks. More hbFGF-MSCs than GFP-MSCs survived and differentiated into vascular endothelial cells (P<0.001); however, their differentiation rates were similar. Moreover, allogenic transplantation of hbFGF-MSCs increased VEGF expression (P=0.008) and microvessel density (P<0.001). Transplantation of hbFGF-expressing MSCs promoted angiogenesis in an in vivo hind limb ischemia model by increasing the survival of transplanted cells that subsequently differentiated into vascular endothelial cells. This study showed the therapeutic potential of combining cell-based therapy with gene therapy to treat ischemic disease.

  5. Bone marrow mesenchymal stem cells overexpressing human basic fibroblast growth factor increase vasculogenesis in ischemic rats

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    J.C. Zhang

    2014-10-01

    Full Text Available Administration or expression of growth factors, as well as implantation of autologous bone marrow cells, promote in vivo angiogenesis. This study investigated the angiogenic potential of combining both approaches through the allogenic transplantation of bone marrow-derived mesenchymal stem cells (MSCs expressing human basic fibroblast growth factor (hbFGF. After establishing a hind limb ischemia model in Sprague Dawley rats, the animals were randomly divided into four treatment groups: MSCs expressing green fluorescent protein (GFP-MSC, MSCs expressing hbFGF (hbFGF-MSC, MSC controls, and phosphate-buffered saline (PBS controls. After 2 weeks, MSC survival and differentiation, hbFGF and vascular endothelial growth factor (VEGF expression, and microvessel density of ischemic muscles were determined. Stable hbFGF expression was observed in the hbFGF-MSC group after 2 weeks. More hbFGF-MSCs than GFP-MSCs survived and differentiated into vascular endothelial cells (P<0.001; however, their differentiation rates were similar. Moreover, allogenic transplantation of hbFGF-MSCs increased VEGF expression (P=0.008 and microvessel density (P<0.001. Transplantation of hbFGF-expressing MSCs promoted angiogenesis in an in vivo hind limb ischemia model by increasing the survival of transplanted cells that subsequently differentiated into vascular endothelial cells. This study showed the therapeutic potential of combining cell-based therapy with gene therapy to treat ischemic disease.

  6. Optimal Route for Mesenchymal Stem Cells Transplantation after Severe Intraventricular Hemorrhage in Newborn Rats.

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    So Yoon Ahn

    Full Text Available Recently, we showed that intracerebroventricular (IC transplantation of human umbilical cord blood (UCB-derived mesenchymal stem cells (MSCs significantly attenuates posthemorrhagic hydrocephalus (PHH and brain damage after severe IVH in newborn rats. This study was performed to determine the optimal route for transplanting MSCs for severe IVH by comparing IC transplantation, intravenous (IV transplantation, and IV transplantation plus mannitol infusion. Severe IVH was induced by injecting 100 uL of blood into each ventricle of Sprague-Dawley rats on postnatal day 4 (P4. After confirming severe IVH with brain magnetic resonance imaging (MRI at P5, human UCB-derived MSCs were transplanted at P6 by an IC route (1×105, an IV route (5×105, or an IV route with mannitol infused. Follow-up brain MRIs and rotarod tests were performed. At P32, brain tissue samples were obtained for biochemical and histological analyses. Although more MSCs localized to the brain after IC than after IV delivery, both methods were equally effective in preventing PHH; attenuating impaired rotarod test; increasing the number of TUNEL-positive cells, inflammatory cytokines, and astrogliosis; and reducing corpus callosal thickness and myelin basic protein expression after severe IVH regardless of mannitol co-infusion. Despite the superior delivery efficacy with IC than with the IV route, both IC and IV transplantation of MSCs had equal therapeutic efficacy in protecting against severe IVH. These findings suggest that the less invasive IV route might be a good alternative for clinically unstable, very preterm infants that cannot tolerate a more invasive IC delivery of MSCs.

  7. Curcumin increases rat mesenchymal stem cell osteoblast differentiation but inhibits adipocyte differentiation

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    Qiaoli Gu

    2012-01-01

    Full Text Available Background: Curcumin is a phenolic natural product isolated from the rhizome of Curcuma longa (turmeric and has effects on bone health and fat formation. The bone marrow mesenchymal stem cells (MSCs are multipotent cells capable of differentiating into osteoblasts and adipocytes. Osteoblast differentiation of MSCs can be a result of upregulation of heme oxygenase (HO-1 expression. Curcumin can potently induce HO-1 expression. Objective: The present study describes the effects of curcumin on rat MSC (rMSCs differentiation into osteoblasts and adipocytes. Materials and Methods: Rat bone marrow MSCs were isolated and treated with or without curcumin. Osteoblast differentiation was confirmed and determined by alkaline phosphatase (ALP activity, mineralized nodule formation, the expression of Runx2 (runt-related transcription factor 2 and osteocalcin. Adipocyte differentiation was determined by Oil red O staining and the expression of peroxisome proliferator-activated receptor-γ 2 (PPARγ2 and CCAAT/enhancer-binding protein (C/EBP α. Results: Curcumin increased ALP activity and osteoblast-specific mRNA expression of Runx2 and osteocalcin when rMSCs were cultured in osteogenic medium. In contrast, curcumin decreased adipocyte differentiation and inhibited adipocyte-specific mRNA expression of PPARγ2 and C/EBPα when rMSCs were cultured in adipogenic medium. HO-1 expression was increased during osteogenic differentiation of rMSCs. Conclusions: These findings demonstrate that curcumin can promote osteogenic differentiation of rMSCs and inhibit adipocyte formation. The effect of curcumin on osteogenic differentiation of rMSCs is correlated with HO-1 expression.

  8. Protective effect of bone marrow mesenchymal stem cells combined with erythropoietin therapy on spinal cord injury rat model

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    Peng Xie; Wen-Hui Ruan

    2016-01-01

    Objective:To study the protective effect of bone marrow mesenchymal stem cells combined with erythropoietin therapy on spinal cord injury rat model.Methods: SD rats were selected as experimental animals, spinal cord injury rat model was built by striking spinal cord with Hatteras Instruments PCI3000, and model rats were divided into control group, bone marrow mesenchymal stem cells (BMSCs) group, erythropoietin (EPO) group and BMSCs combined with EPO group according to different treatment methods. Then number of apoptotic cells in spinal cord tissue, contents of neural markers and neurotrophic factors as well as expression of apoptosis and injury molecules was detected.Results:Number of apoptotic cells as well as mRNA contents of Caspase-3 and c-fos of BMSCs group, EPO group and BMSCs+EPO group was lower than those of control group, and number of apoptotic cells as well as mRNA contents of Caspase-3 and c-fos of BMSCs+EPO group were lower than those of BMSCs group and EPO group; mRNA contents of NF-200 and MBP as well as protein contents of NGF and BDNF in spinal cord tissue of BMSCs group, EPO group and BMSCs+EPO group were higher than those of control group, and mRNA contents of NF-200 and MBP as well as protein contents of NGF and BDNF in spinal cord tissue of BMSCs+EPO group were higher than those of BMSCs group and EPO group.Conclusions:Bone marrow mesenchymal stem cells combined with erythropoietin therapy can inhibit cell apoptosis in the injured spinal cord tissue, increase neurotrophic factor levels and inhibit apoptosis and injury molecule expression; it has protective effect on spinal cord injury.

  9. Low power laser and LED irradiation effect on proliferation and differentiation of Wistar rats mesenchymal stem cells

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    Mancera, Diana; Solarte, Efrain; Fierro, Leonardo; Criollo, William

    2013-11-01

    It has been demonstrated that appropriately cultured and stimulated mesenchymal cells, can give rise to cells of all tissues of the body. We evaluate the cell proliferation and differentiation induced by low power light irradiation in cell cultures of mesenchymal cells, isolated and previously characterized, from Wistar rats. Roche® XTT and LDH tests were used to assess proliferation and cytotoxicity. Cellular differentiation was determined by optical microscopy and using specific fluorescent markers. We report laser cellular proliferation enhancement by 532 and 473 nm, and the best cell culture response by a dose of 2 Jcm-2. Although a three day irradiation protocol the cultures grown and no cytotoxicity was detected. Cellular differentiation occurred, and the production of cardiomyocytes was promoted by the cell proliferation stimulated by low power laser irradiation.

  10. In vivo transplantation of bone marrow mesenchymal stem cells accelerates repair of injured gastric mucosa in rats

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    CHANG Qing; YAN Li; WANG Chang-zheng; ZHANG Wen-hui; HU Ya-zhuo; WU Ben-yan

    2012-01-01

    Background Adult stem cells provide a promising alternative for the treatment of injured tissues.We aimed to investigate the effect of in vivo transplantation of bone marrow mesenchymal stem cells (BMMSCs) on injured gastric mucosa in rats.Methods The gastric ulcer in rats was induced by indomethacin.BMMSCs from male rats,labeled with the fluorescent cell linker 5,6-carboxyfluorescein diacetate succinimidyl ester (CFDA SE),were transplanted into the female rats via tail vein injection.The healing process of gastric ulcers was monitored by HE staining.The protein levels of vascular endothelial growth factor (VEGF) and the epidermal growth factor receptor (EGFR) in the injured gastric mucosa were determined by immunohistochemistry.Results At 48 and 72 hours after BMMSCs transplantation,the CFDA SE labeled cells were found scattered in the injured gastric mucosa,but not in the gastric mucosa of control rats.At 72 hours after BMMSCs transplantation,the mean ulcer index was 12.67±2.16 in the BMMSCs transplanted group and 17.33±1.97 in vehicle-treated controls (P <0.01).Both VEGF and EGFR protein expression levels were significantly higher in the gastric section from the rats that received BMMSCs transplantation as compared to rats without BMMSCs transplantation.Conclusion Autologous BMMSCs transplantation can accelerate gastric ulcer healing in injured gastric mucosa in a rodent model.

  11. Human umbilical cord mesenchymal stem cells transplantation promotes cutaneous wound healing of severe burned rats.

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    Lingying Liu

    Full Text Available BACKGROUND: Severe burns are a common and highly lethal trauma. The key step for severe burn therapy is to promote the wound healing as early as possible, and reports indicate that mesenchymal stem cell (MSC therapy contributes to facilitate wound healing. In this study, we investigated effect of human umbilical cord MSCs (hUC-MSCs could on wound healing in a rat model of severe burn and its potential mechanism. METHODS: Adult male Wistar rats were randomly divided into sham, burn, and burn transplanted hUC-MSCs. GFP labeled hUC-MSCs or PBS was intravenous injected into respective groups. The rate of wound closure was evaluated by Image Pro Plus. GFP-labeled hUC-MSCs were tracked by in vivo bioluminescence imaging (BLI, and human-specific DNA expression in wounds was detected by PCR. Inflammatory cells, neutrophils, macrophages, capillaries and collagen types I/III in wounds were evaluated by histochemical staining. Wound blood flow was evaluated by laser Doppler blood flow meter. The levels of proinflammatory and anti-inflammatory factors, VEGF, collagen types I/III in wounds were analyzed using an ELISA. RESULTS: We found that wound healing was significantly accelerated in the hUC-MSC therapy group. The hUC-MSCs migrated into wound and remarkably decreased the quantity of infiltrated inflammatory cells and levels of IL-1, IL-6, TNF-α and increased levels of IL-10 and TSG-6 in wounds. Additionally, the neovascularization and levels of VEGF in wounds in the hUC-MSC therapy group were markedly higher than those in other control groups. The ratio of collagen types I and III in the hUC-MSC therapy group were markedly higher than that in the burn group at indicated time after transplantation. CONCLUSION: The study suggests that hUC-MSCs transplantation can effectively improve wound healing in severe burned rat model. Moreover, these data might provide the theoretical foundation for the further clinical application of hUC-MSC in burn areas.

  12. Rat bone marrow mesenchymal stem cells differentiate into hepatocytes in vitro

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    Xin-Qin Kang; Wei-Jin Zang; Tu-Sheng Song; Xiao-Li Xu; Xiao-Jiang Yu; Dong-Ling Li; Ke-Wei Meng; Sheng-Li Wu; Zhi-Ying Zhao

    2005-01-01

    AIM: To investigate the mechanism and regulation of differentiation from bone marrow mesenchymal stem cells (MSCs) into hepatocytes and to find a new source of celltypes for therapies of hepatic diseases. METHODS: MSCs were isolated by combining gradient density centrifugation with plastic adherence. The cells were cultured in osteogenic or adipogenic differentiation medium and determined by histochemical staining. MSCs were plated in plastic culture flasks that were not coated with components of extracellular matrix (ECM). When MSCs reached 70% confluence, they were cultured in low glucose Dulbecco's modified Eagle's medium supplemented with 10 mL/L fetal bovine serum, 20 ng/mL hepatocyte growth factor (HGF) and 10 ng/mL fibroblast growth factor-4 (FGF-4). The medium was changed every 3 d and stored for albumin, alpha-fetoprotein (AFP) and urea assay. Glycogen store of hepatocytes was determined by periodic acid-Schiff staining.RESULTS: By combining gradient density centrifugation with plastic adherence, we isolated a homogeneous population of cells from rat bone marrow and differentiated them into osteocytes and adipocytes. When MSCs were cultured withFGF-4 and HGF, approximately 56.6% of cells became smallround and epithelioid on d 24 by morphology. Compared with the control, levels of AFP increased significantly from d 12 to 15.5±1.4 μg/L (t = 2.31, P<0.05) in MSCs cultured with FGF-4and HGF, and were higher (46.2±1.5 μg/L)ond 21 (t = 41.926, P<0.01), then decreased to 24.8±2.2 μg/L on d 24 (t = 10.345, P<0.01). Albumin increased significantly on d 21 (t= 3.325, P<0.01) to 1.4±0.2 μg/mL,and to 2.1±0.7 μg/mL on d 24 (t= 3.646, P<0.01). Urea(2.3±0.4 mmol/L) was first detected on d 21 (t = 6.739, P<0.01), and continued to increase to 2.6±0.9 mmol/Lon d 24 (t= 4.753, P<0.01). Glycogen storage was first seen on d 21.CONCLUSION: The method combining gradient density centrifugation with plastic adherence can isolate MSCs. Rat MSCs may be

  13. Use of Rats Mesenchymal Stem Cells Modified with mHCN2 Gene to Create Biologic Pacemakers

    Institute of Scientific and Technical Information of China (English)

    马金; 张存泰; 黄深; 王国强; 全小庆

    2010-01-01

    The possibility of rats mesenchymal stem cells (MSCs) modified with murine hyperpolarization-activated cyclic nucleotide-gated 2 (mHCN2) gene as biological pacemakers in vitro was studied. The cultured MSCs were transfected with pIRES2-EGFP plasmid carrying enhanced green fluorescent protein (EGFP) gene and mHCN2 gene. The identification using restriction enzyme and sequencing indicated that the mHCN2 gene was inserted to the pIRES2-EGFP. Green fluorescence could be seen in MSCs after transfection for 24-48...

  14. Cognitive improvement following transvenous adipose-derived mesenchymal stem cell transplantation in a rat model of traumatic brain injury

    Institute of Scientific and Technical Information of China (English)

    Dongfei Li; Chun Yang; Rongmei Qu; Huiying Yang; Meichun Yu; Hui Tao; Jingxing Dai; Lin Yuan

    2011-01-01

    The effects of adipose-derived mesenchymal stem cell (ADMSC) transplantation for the repair of traumatic brain injury remain poorly understood. The present study observed neurological functional changes in a rat model of traumatic brain injury following ADMSC transplantation via the tail vein.Cell transplants were observed in injured cerebral cortex, and expression of brain-derived nerve growth factor was significantly increased in the injured hippocampus following transplantation. Results demonstrated that transvenous ADMSC transplants migrated to the injured cerebral cortex and significantly improved cognitive function.

  15. Collagen-GAG Scaffolds Grafted Onto Myocardial Infarcts in a Rat Model:A Delivery Vehicle for Mesenchymal Stem Cells

    Institute of Scientific and Technical Information of China (English)

    Z.; XIANG; R.; LIAO; M.; KELLY; M.; SPECTOR

    2005-01-01

    1 IntroductionThe objective of the present study was to investigate the response of rat myocardial scar tissue to type I collagen-glycosaminoglycan (GAG) tissue engineering scaffolds, and to assess the feasibility of using a collagen-GAG scaffold as a delivery vehicle for bone marrow-derived mesenchymal stem cells. The benefits of employing the collagen-GAG scaffold for this application include the following:(1) the large surface area of the three-dimensional sponge-like material allows for the delivery of ...

  16. Theobromine Upregulates Osteogenesis by Human Mesenchymal Stem Cells In Vitro and Accelerates Bone Development in Rats.

    Science.gov (United States)

    Clough, Bret H; Ylostalo, Joni; Browder, Elizabeth; McNeill, Eoin P; Bartosh, Thomas J; Rawls, H Ralph; Nakamoto, Tetsuo; Gregory, Carl A

    2017-03-01

    Theobromine (THB) is one of the major xanthine-like alkaloids found in cacao plant and a variety of other foodstuffs such as tea leaves, guarana and cola nuts. Historically, THB and its derivatives have been utilized to treat cardiac and circulatory disorders, drug-induced nephrotoxicity, proteinuria and as an immune-modulator. Our previous work demonstrated that THB has the capacity to improve the formation of hydroxyl-apatite during tooth development, suggesting that it may also enhance skeletal development. With its excellent safety profile and resistance to pharmacokinetic elimination, we reasoned that it might be an excellent natural osteoanabolic supplement during pregnancy, lactation and early postnatal growth. To determine whether THB had an effect on human osteoprogenitors, we subjected primary human bone marrow mesenchymal stem cells (hMSCs) to osteogenic assays after exposure to THB in vitro and observed that THB exposure increased the rate of osteogenesis and mineralization by hMSCs. Moreover, THB exposure resulted in a list of upregulated mRNA transcripts that best matched an osteogenic tissue expression signature as compared to other tissue expression signatures archived in several databases. To determine whether oral administration of THB resulted in improved skeletal growth, we provided pregnant rats with chow supplemented with THB during pregnancy and lactation. After weaning, offspring received THB continuously until postnatal day 50 (approximately 10 mg kg(-1) day(-1)). Administration of THB resulted in neonates with larger bones, and 50-day-old offspring accumulated greater body mass, longer and thicker femora and superior tibial trabecular parameters. The accelerated growth did not adversely affect the strength and resilience of the bones. These results indicate that THB increases the osteogenic potential of bone marrow osteoprogenitors, and dietary supplementation of a safe dose of THB to expectant mothers and during the postnatal period

  17. Potential Spermatogenesis Recovery with Bone Marrow Mesenchymal Stem Cells in an Azoospermic Rat Model

    Directory of Open Access Journals (Sweden)

    Deying Zhang

    2014-07-01

    Full Text Available Non-obstructive azoospermia is the most challenging type of male infertility. Stem cell based therapy provides the potential to enhance the recovery of spermatogenesis following cancer therapy. Bone marrow-derived mesenchymal stem cells (BMSCs possess the potential to differentiate or trans-differentiate into multi-lineage cells, secrete paracrine factors to recruit the resident stem cells to participate in tissue regeneration, or fuse with the local cells in the affected region. In this study, we tested whether spermatogenically-induced BMSCs can restore spermatogenesis after administration of an anticancer drug. Allogeneic BMSCs were co-cultured in conditioned media derived from cultured testicular Sertoli cells in vitro, and then induced stem cells were transplanted into the seminiferous tubules of a busulfan-induced azoospermatic rat model for 8 weeks. The in vitro induced BMSCs exhibited specific spermatogonic gene and protein markers, and after implantation the donor cells survived and located at the basement membranes of the recipient seminiferous tubules, in accordance with what are considered the unique biological characteristics of spermatogenic stem cells. Molecular markers of spermatogonial stem cells and spermatogonia (Vasa, Stella, SMAD1, Dazl, GCNF, HSP90α, integrinβ1, and c-kit were expressed in the recipient testis tissue. No tumor mass, immune response, or inflammatory reaction developed. In conclusion, BMSCs might provide the potential to trans-differentiate into spermatogenic-like-cells, enhancing endogenous fertility recovery. The present study indicates that BMSCs might offer alternative treatment for the patients with azoospermatic infertility after cancer chemotherapy.

  18. Differentiation potential of bone marrow mesenchymal stem cells into retina in normal and laser-injured rat eye

    Institute of Scientific and Technical Information of China (English)

    ZHANG Jie; SHAN Qing; MA Ping; JIANG Yanming; CHEN Peng; WEN Jingxia; ZHOU You; QIAN Huanwen; PEI Xuetao

    2004-01-01

    Bone marrow mesenchymal stem cells (MSCs) can develop into hematopoietic and mesenchymal lineages but have not been known to participate in the production of retina. Here we report that bone marrow mesenchymal stem cells, after being subretinally transplanted into normal or Nd: YAG laser-injured rat eye, can integrate into RPE layer, photoreceptor layer, bipolar cell layer and ganglion layer. DAPI-labeling detection was used to trace the origin of the repopulating cells. DAPI fluorescence was used to identify retina cells of bone marrow origin 10, 20, 35 and 50 days after transplantation. No formation of rosettes was found but some random cells were found at the end of the observation. MSCs-originated cells spread more widely in the injured retinas than in the normal ones. Immunohistochemical detection showed that though the cells could express neuronal nuclei (NeuN), neuron specific enolase (NSE), glial fibrillary acidic protein (GFAP) and cytokeratin (CK), the proteins expression in the injured transplantation group was abnormal in some region compared with that in the normal transplantation group. Electroretinogram (ERG) showed that ERG-b wave of the injured transplantation group is significantly higher than that of the two laser-injured control groups. These results suggest that a proportion of MSCs can differentiate into retina-like structure in vivo and the differentiation differs in normal and laser-injured retinas.

  19. Effects of strontium on proliferation and differentiation of rat bone marrow mesenchymal stem cells

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    Li, Yunfeng; Li, Jihua; Zhu, Songsong; Luo, En; Feng, Ge; Chen, Qianming [State Key Laboratory of Oral Diseases, West China College of Stomatology, Sichuan University, No. 14, Section 3, Southern Renmin Road, Chengdu 610041 (China); Hu, Jing, E-mail: drhu@vip.sohu.com [State Key Laboratory of Oral Diseases, West China College of Stomatology, Sichuan University, No. 14, Section 3, Southern Renmin Road, Chengdu 610041 (China)

    2012-02-24

    Highlights: Black-Right-Pointing-Pointer Strontium ranelate (SrR) inhibits proliferation of BMMSCs. Black-Right-Pointing-Pointer SrR increases osteoblastic but decreases adipocytic differentiation of BMMSCs. Black-Right-Pointing-Pointer SrR increases expression of Runx2, BSP and OCN by BMMSCs in osteogenic medium. Black-Right-Pointing-Pointer SrR decreases expression of PPAR{gamma}, aP2/ALBP and LPL by BMMSCs in adipogenic medium. -- Abstract: Strontium ranelate (SrR) was an effective anti-osteoporotic drug to increase bone formation and decrease bone resorption. However, reports about the effect of SR on osteoblastic and adipocytic differentiation from bone marrow mesenchymal stem cells (BMMSCs) are limited. The purpose of this study is to evaluate whether SrR affects the ability of BMMSCs to differentiate into osteoblasts or adipocytes. Rat BMMSCs were identified by flow cytometry and exposed to SR (0.1 and 1.0 mM Sr{sup 2+}) under osteogenic or adipogenic medium for 1 and 2 weeks. The proliferation and differentiation of BMMSCs were analyzed by MTT, alkaline phosphatase (ALP), Oil red O staining, quantitative real-time RT-PCR and Western blot assays. SrR significantly inhibited the proliferation, increased osteoblastic but decreased adipocytic differentiation of rat BMMSCs dose-dependently. In osteogenic medium, SrR increased the expression of ALP, the mRNA levels of Cbfa1/Runx2, bone sialoprotein, and osteocalcin by RT-PCR, and the protein levels of Cbfa1/Runx2 by Western blot. In adipogenic medium, SrR decreased the mRNA levels of PPAR{gamma}2, adipocyte lipid-binding protein 2 (aP2/ALBP), and lipoprotein lipase (LPL) by RT-PCR, and the protein expression of PPAR{gamma} in Western blot analysis. These results indicated that the effects of SrR to promote osteoblastic but inhibit adipocytic differentiation of BMMSCs might contribute to its effect on osteoporosis treatment.

  20. Therapeutic efficacy of intranasally delivered mesenchymal stem cells in a rat model of Parkinson disease.

    Science.gov (United States)

    Danielyan, Lusine; Schäfer, Richard; von Ameln-Mayerhofer, Andreas; Bernhard, Felix; Verleysdonk, Stephan; Buadze, Marine; Lourhmati, Ali; Klopfer, Tim; Schaumann, Felix; Schmid, Barbara; Koehle, Christoph; Proksch, Barbara; Weissert, Robert; Reichardt, Holger M; van den Brandt, Jens; Buniatian, Gayane H; Schwab, Matthias; Gleiter, Christoph H; Frey, William H

    2011-02-01

    Safe and effective cell delivery remains one of the main challenges in cell-based therapy of neurodegenerative disorders. Graft survival, sufficient enrichment of therapeutic cells in the brain, and avoidance of their distribution throughout the peripheral organs are greatly influenced by the method of delivery. Here we demonstrate for the first time noninvasive intranasal (IN) delivery of mesenchymal stem cells (MSCs) to the brains of unilaterally 6-hydroxydopamine (6-OHDA)-lesioned rats. IN application (INA) of MSCs resulted in the appearance of cells in the olfactory bulb, cortex, hippocampus, striatum, cerebellum, brainstem, and spinal cord. Out of 1 × 10⁶ MSCs applied intranasally, 24% survived for at least 4.5 months in the brains of 6-OHDA rats as assessed by quantification of enhanced green fluorescent protein (EGFP) DNA. Quantification of proliferating cell nuclear antigen-positive EGFP-MSCs showed that 3% of applied MSCs were proliferative 4.5 months after application. INA of MSCs increased the tyrosine hydroxylase level in the lesioned ipsilateral striatum and substantia nigra, and completely eliminated the 6-OHDA-induced increase in terminal deoxynucleotidyl transferase (TdT)-mediated 2'-deoxyuridine, 5'-triphosphate (dUTP)-biotin nick end labeling (TUNEL) staining of these areas. INA of EGFP-labeled MSCs prevented any decrease in the dopamine level in the lesioned hemisphere, whereas the lesioned side of the control animals revealed significantly lower levels of dopamine 4.5 months after 6-OHDA treatment. Behavioral analyses revealed significant and substantial improvement of motor function of the Parkinsonian forepaw to up to 68% of the normal value 40-110 days after INA of 1 × 10⁶ cells. MSC-INA decreased the concentrations of inflammatory cytokines-interleukin-1β (IL-1β), IL-2, -6, -12, tumor necrosis factor (TNF), interferon-γ (IFN-γ, and granulocyte-macrophage colony-stimulating factor (GM-CSF)-in the lesioned side to their

  1. Effects of human mesenchymal stem cell transplantation in the bilateral corpus striatum in a rat model of Tourette's syndrome

    Institute of Scientific and Technical Information of China (English)

    Xiumei Liu; Yuwei Wang; Mingji Yi

    2010-01-01

    Tourette's syndrome is treated by behavioral or pharmacological therapy.However,patients with malignant Tourette's syndrome also exhibit life-threatening symptoms,which are unresponsive to conservative treatments or neurosurgical procedures,such as deep brain stimulation.In recent years,mesenchymal stem cells(MSCs)have shown therapeutic potential in many neurological diseases.Therefore,the present study proposed to use MSC transplantation as a novel therapy for Tourette's syndrome.Stereotypic behaviors in Tourette's syndrome rats decreased significantly at21 days after human MSCs transplantation into the striatum.Immunohistochemistry analyses revealed survival of transplanted human MSCs and differentiation into neurons and astrocytes in the rat brain.Results suggest that intrastriatal transplantation of human MSCs could provide therapeutic potential for Tourette's syndrome.

  2. Effect of Rougan Huaqian granules combined with human mesenchymal stem cell transplantation on liver fibrosis in cirrhosis rats

    Institute of Scientific and Technical Information of China (English)

    Zhen-Chang Wang; Shan Yang; Jing-Jing Huang; Song-Lin Chen; Quan-Qiang Li; Yuan Li

    2014-01-01

    Objective:To observe the effect ofRouganHuaqian granules combined with human mesenchymal stem cell(hMSC) transplantation on the liver fibrosis in carbon tetrachloride-induced cirrhosis rats.Methods:SixtySD rats were randomly divided into five groups.The rats in control group received intraperitoneal injection of saline, while those in model control group, treatment groupA, groupB and groupC received intraperitoneal injection of carbon tetrachloride oily solution to induce liver cirrhosis within8 weeks.Then, the rats in the model control group, treatment groupA, treatment groupB, treatment groupC received vein tail injection of saline, RouganHuaqian granules, hMSC suspension andRouganHuaqian granules combined with hMSC suspension.Results:The treatment groups had significantly different liverfunction(AST levels), liver fibrosis index(laminin andHA), hepatic sinusoidal wallsα-smooth muscle actin,Ⅳ collagen and laminin protein expression andⅠ,Ⅲ collagen from the model group(P<0.05). The transplanted cells showed human hepatocyte-like cells differentiation trend in the liver. Conclusions:TheRouganHuaqian granules combined with hMSC transplantation can alleviate liver fibrosis in cirrhosis rats.

  3. In Vivo Tracking of Systemically Administered Allogeneic Bone Marrow Mesenchymal Stem Cells in Normal Rats through Bioluminescence Imaging

    Directory of Open Access Journals (Sweden)

    Juan Cao

    2016-01-01

    Full Text Available Recently, mesenchymal stem cells (MSCs are increasingly used as a panacea for multiple types of disease short of effective treatment. Dozens of clinical trials published demonstrated strikingly positive therapeutic effects of MSCs. However, as a specific agent, little research has focused on the dynamic distribution of MSCs after in vivo administration. In this study, we track systemically transplanted allogeneic bone marrow mesenchymal stem cells (BMSCs in normal rats through bioluminescence imaging (BLI in real time. Ex vivo organ imaging, immunohistochemistry (IHC, and RT-PCR were conducted to verify the histological distribution of BMSCs. Our results showed that BMSCs home to the dorsal skin apart from the lungs and kidneys after tail vein injection and could not be detected 14 days later. Allogeneic BMSCs mainly appeared not at the parenchymatous organs but at the subepidermal connective tissue and adipose tissue in healthy rats. There were no significant MSCs-related adverse effects except for transient decrease in neutrophils. These findings will provide experimental evidences for a better understanding of the biocharacteristics of BMSCs.

  4. The Effect of Bone Marrow-Derived Mesenchymal Stem Cells and Their Conditioned Media Topically Delivered in Fibrin Glue on Chronic Wound Healing in Rats

    OpenAIRE

    Mehanna, Radwa A.; Iman Nabil; Noha Attia; Bary, Amany A.; Razek, Khalid A.; Ahmed, Tamer A. E.; Fatma Elsayed

    2015-01-01

    Bone marrow-derived mesenchymal stem cells (BM-MSCs) represent a modern approach for management of chronic skin injuries. In this work, we describe BM-MSCs application versus their conditioned media (CM) when delivered topically admixed with fibrin glue to enhance the healing of chronic excisional wounds in rats. Fifty-two adult male rats were classified into four groups after induction of large-sized full-thickness skin wound: control group (CG), fibrin only group (FG), fibrin + MSCs group (...

  5. Recovery of neurological function of ischemic stroke by application of conditioned medium of bone marrow mesenchymal stem cells derived from normal and cerebral ischemia rats

    OpenAIRE

    2014-01-01

    Background Several lines of evidence have demonstrated that bone marrow-derived mesenchymal stem cells (BM-MSC) release bioactive factors and provide neuroprotection for CNS injury. However, it remains elusive whether BM-MSC derived from healthy donors or stroke patients provides equal therapeutic potential. The present work aims to characterize BM-MSC prepared from normal healthy rats (NormBM-MSC) and cerebral ischemia rats (IschBM-MSC), and examine the effects of their conditioned medium (C...

  6. Salt-induced epithelial-to-mesenchymal transition in Dahl salt-sensitive rats is dependent on elevated blood pressure

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Y.; Mu, J.J.; Liu, F.Q.; Ren, K.Y.; Xiao, H.Y. [Xi' an Jiaotong University, Medical College, First Affiliated Hospital, Cardiovascular Department, Xi' an, China, Cardiovascular Department, First Affiliated Hospital, Medical College, Xi' an Jiaotong University, Xi' an (China); Ministry of Education, Key Laboratory of Environment and Genes Related to Diseases, Xi' an, China, Key Laboratory of Environment and Genes Related to Diseases, Ministry of Education, Xi' an (China); Yang, Z. [Xi' an Jiaotong University, Medical College, First Affiliated Hospital, Department of Pathology, Xi' an, China, Department of Pathology, First Affiliated Hospital, Medical College, Xi' an Jiaotong University, Xi' an (China); Yuan, Z.Y. [Xi' an Jiaotong University, Medical College, First Affiliated Hospital, Cardiovascular Department, Xi' an, China, Cardiovascular Department, First Affiliated Hospital, Medical College, Xi' an Jiaotong University, Xi' an (China); Ministry of Education, Key Laboratory of Environment and Genes Related to Diseases, Xi' an, China, Key Laboratory of Environment and Genes Related to Diseases, Ministry of Education, Xi' an (China)

    2014-03-03

    Dietary salt intake has been linked to hypertension and cardiovascular disease. Accumulating evidence has indicated that salt-sensitive individuals on high salt intake are more likely to develop renal fibrosis. Epithelial-to-mesenchymal transition (EMT) participates in the development and progression of renal fibrosis in humans and animals. The objective of this study was to investigate the impact of a high-salt diet on EMT in Dahl salt-sensitive (SS) rats. Twenty-four male SS and consomic SS-13{sup BN} rats were randomized to a normal diet or a high-salt diet. After 4 weeks, systolic blood pressure (SBP) and albuminuria were analyzed, and renal fibrosis was histopathologically evaluated. Tubular EMT was evaluated using immunohistochemistry and real-time PCR with E-cadherin and alpha smooth muscle actin (α-SMA). After 4 weeks, SBP and albuminuria were significantly increased in the SS high-salt group compared with the normal diet group. Dietary salt intake induced renal fibrosis and tubular EMT as identified by reduced expression of E-cadherin and enhanced expression of α-SMA in SS rats. Both blood pressure and renal interstitial fibrosis were negatively correlated with E-cadherin but positively correlated with α-SMA. Salt intake induced tubular EMT and renal injury in SS rats, and this relationship might depend on the increase in blood pressure.

  7. MR tracking of SPIO-labeled mesenchymal stem cells in rats with liver fibrosis could not monitor the cells accurately.

    Science.gov (United States)

    Zhou, Bin; Li, Dan; Qian, Jiesheng; Li, Zhengran; Pang, Pengfei; Shan, Hong

    2015-01-01

    Our previous study showed that in vivo magnetic resonance (MR) imaging is effective in tracking superparamagnetic iron oxide (SPIO)-labeled bone marrow mesenchymal stem cells (BMSCs) in rats with liver fibrosis. SPIO-labeling-induced signal reduction on MR images was completely reversed within 15 days after transplantation. It is still unclear whether the signal changes in MR imaging could reflect the number of transplanted cells in the liver. In the present study, BMSCs of male rats were doubly labeled with enhanced green fluorescent protein (EGFP) and SPIO and injected intravascularly into female rats with liver fibrosis. At different time points after injection, MR imaging was performed. The distribution of SPIO particles and EGFP-positive cells was determined by Prussian blue staining and EGFP immunohistochemistry, respectively. The distribution of transplanted BMSCs in various organs was assessed by detection of the SRY gene using real-time quantitative PCR. At 15 days post transplantation, the numbers of transplanted cells were significantly decreased in the lung, kidney, spleen and muscle, but not liver and heart, in comparison with those at 7 days after transplantation. EGFP staining-positive cells were observed in the liver intralobular parenchyma, while Prussian blue staining was negative at 42 days after transplantation. Taken together, SPIO particles and EGFP-labeled BMSCs show a different tissue distribution pattern in rats with liver fibrosis after a long-term period of monitoring. SPIO-based MR imaging may not be suitable for long-term tracking of transplanted BMSCs in vivo.

  8. Effect of mesenchymal stem cells on anti-Thy1,1 induced kidney injury in albino rats

    Institute of Scientific and Technical Information of China (English)

    Saber Sakr; Laila Rashed; Waheba Zarouk; Rania El-Shamy

    2013-01-01

    Objective: To evaluate the effect of mesenchymal stem cells (MSCs) in rats with anti-Thy1,1 nephritis. Methods: Female albino rats were divided into three groups, control group, anti-Thy1,1 group and treatment with i.v. MSCs group. MSCs were derived from bone marrow of male albino rats, Y-chromosome gene was detected by polymerase chain reaction in the kidney. Serum urea and creatinine were estimated for all groups. Kidney of all studied groups was examined histologically and histochemically (total carbohydrates and total proteins). DNA fragmentation and expression of α-SMA were detected. Results:Kidney of animals injected with anti-Thy1,1 showed inflammatory leucocytic infiltration, hypertrophied glomeruli, tubular necrosis and congestion in the renal blood vessels. The kidney tissue also showed reduction of carbohydrates and total proteins together with increase in apoptosis and in expression ofα-SMA. Moreover, the levels of urea and creatinine were elevated. Treating animals with MSCs revealed that kidney tissue displayed an improvement in the histological and histochemical changes. Apoptosis and α-SMA expression were decreased, and the levels of urea and creatinine decreased. Conclusions:The obtained results demonstrated the potential of MSCs to ameliorate the structure and function of the kidney in rats with anti-Thy1,1 nephritis possibly through the release of paracrine growth factor(s).

  9. Human embryonic mesenchymal stem cell-derived conditioned medium rescues kidney function in rats with established chronic kidney disease.

    Directory of Open Access Journals (Sweden)

    Arianne van Koppen

    Full Text Available Chronic kidney disease (CKD is a major health care problem, affecting more than 35% of the elderly population worldwide. New interventions to slow or prevent disease progression are urgently needed. Beneficial effects of mesenchymal stem cells (MSC have been described, however it is unclear whether the MSCs themselves or their secretome is required. We hypothesized that MSC-derived conditioned medium (CM reduces progression of CKD and studied functional and structural effects in a rat model of established CKD. CKD was induced by 5/6 nephrectomy (SNX combined with L-NNA and 6% NaCl diet in Lewis rats. Six weeks after SNX, CKD rats received either 50 µg CM or 50 µg non-CM (NCM twice daily intravenously for four consecutive days. Six weeks after treatment CM administration was functionally effective: glomerular filtration rate (inulin clearance and effective renal plasma flow (PAH clearance were significantly higher in CM vs. NCM-treatment. Systolic blood pressure was lower in CM compared to NCM. Proteinuria tended to be lower after CM. Tubular and glomerular damage were reduced and more glomerular endothelial cells were found after CM. DNA damage repair was increased after CM. MSC-CM derived exosomes, tested in the same experimental setting, showed no protective effect on the kidney. In a rat model of established CKD, we demonstrated that administration of MSC-CM has a long-lasting therapeutic rescue function shown by decreased progression of CKD and reduced hypertension and glomerular injury.

  10. Effect of mesenchymal stem cells transplantation on glycaemic profile & their localization in streptozotocin induced diabetic Wistar rats

    Directory of Open Access Journals (Sweden)

    Shobhit Bhansali

    2015-01-01

    Full Text Available Background & objectives: Bone marrow is a rich source of adult stem cells that can differentiate into various cell types. Administration of mesenchymal stem cells (MSCs in irradiated diabetic rat model has transiently shown to decrease blood glucose level. This study examines the effect of high dose and multiple injections of MSCs on glycemic profile, their localization and regeneration of islet in diabetic Wistar rat. Methods: The study was carried out in male Wistar rats categorized into three groups (n=6, in each group: Group 1 as control, group 2 streptozotocin (STZ (50 mg/kg induced diabetic group and group 3 experimental group; 5-bromo-2-deoxyuridine (BrdU labelled allogenic MSCs were injected in the non-irradiated diabetic rat of the experimental group through tail vein. The blood glucose profile was subsequently monitored at regular intervals. Rats were sacrificed on day 45 and pancreas was examined for localization of BrdU labelled stem cells by immunofluorescence and islet-neogenesis by immunohistochemistry . Results: There was a significant reduction in blood glucose level after administration of MSCs in the experimental group (P<0.001. The presence of BrdU labelled MSCs in islet suggested their localization in the pancreas. Co-expression of anti-BrdU and anti-insulin antibody indicated trans-differentiation / fusion into insulin producing cells evidenced by significant increase in total number of islet (P=0.004 and insulin positive cells ( P<0.0001 in experimental group. Interpretation & conclusions: Our results showed that the MSCs administration in non-irradiated diabetic Wistar rat reduced hyperglycaemia and was accompanied by increased islet-neogengesis, possibly through trans- differentiation/fusion.

  11. A novel rat fibrosarcoma cell line from transformed bone marrow-derived mesenchymal stem cells with maintained in vitro and in vivo stemness properties.

    Science.gov (United States)

    Wang, Meng-Yu; Nestvold, Janne; Rekdal, Øystein; Kvalheim, Gunnar; Fodstad, Øystein

    2017-02-09

    Increasing evidence suggests a possible relationship between mesenchymal stem cells (MSCs) and sarcoma. MSCs are hypothesized to be the cells initiating sarcomagenesis, and cancer stem cells (CSCs) sharing features of MSCs have been identified in sarcomas. Here, we report on the characteristics of a bone marrow-derived rat mesenchymal stem cell line that spontaneously transformed in long-term culture. The rat transformed mesenchymal stem cells (rTMSCs) produced soft-tissue fibrosarcomas in immunocompromised mice and immunocompetent rats. In vitro, the rTMSCs displayed increased proliferation capacity compared to the untransformed cell line. The transformed MSCs maintained the mesenchymal phenotype by expression of the stem cell marker CD 90 and the lack of hematopoietic and endothelial markers. Cytogenetic analysis detected trisomy 6 in the rTMSCs. Side population (SP) isolation and tumorsphere cultivation of the transformed cells confirmed the presence of CSCs among the rTMSCs. Importantly, the rTMSCs retained their differentiation capacity towards osteogenic and adipogenic lineages. This transformed MSC-based cell line may be valuable in examining the balance in a mixed cell population between cancer stem cell properties and the ability to differentiate to specific non-transformed cell populations. Moreover, it may also be a useful tool to evaluate the efficacy of novel targeted immunotherapies in vivo.

  12. Increased stromal-cell-derived factor 1 enhances the homing of bone marrow derived mesenchymal stem cells in dilated cardiomyopathy in rats

    Institute of Scientific and Technical Information of China (English)

    ZHOU Yan-li; Michael Fu; ZHANG Hai-feng; LI Xin-li; DI Ruo-min; YAO Wen-ming; LI Dian-fu; FENG Jian-lin; HUANG Jun; CAO Ke-jiang

    2010-01-01

    Background Stem cell transplantation has been shown to have beneficial effects on dilated cardiomyopathy. However,mechanism for stem cell homing to cardiac tissue in dilated cardiomyopathy has not yet been elucidated.Methods Mesenchymal stem cells were obtained from rat bone marrow, expanded in vitro, and labeled with 99mTc.Cardiomyopathy model was induced by doxorubicin in rats. 99mTc labeled cells were infused into the left ventricles in cardiomyopathy and control rats. Sixteen hours after injection, animals were sacrificed and different tissues were harvested to measure specific radioactivity. By use of real-time polymerase chain reaction and immunohistochemistry,Mrna and protein expressions for stromal-cell-derived factor 1 in cardiac tissue were measured.Results Labeling efficiency of mesenchymal stem cells was (70.0±11.2)%. Sixteen hours after mesenchymal stem cell transplantation, the heart-to-muscle radioactivity ratio was increased significantly in cardiomyopathy hearts as compared to control hearts. Both Mrna and rotein expressions of stromal-cell-derived factor 1 were up-regulated in cardiomyopathy hearts as compared with control hearts.Conclusion In dilated cardiomyopathy induced by doxorubicin up-regulated expression of stromal-cell-derived factor 1in heart may induce mesenchymal stem cells home to the heart.

  13. The effects of bone marrow mesenchymal stromal cells transplantation after mannitol pretreatment on behavioral performance and synaptophysin expression in the CA3 region in hippocampus of vascular dementia rats

    Institute of Scientific and Technical Information of China (English)

    农伟东

    2013-01-01

    Objective To investigate the effects of bone marrow mesenchymal stromal cells (BMSCs) transplantation after mannitol pretreatment on behavioral performance and synaptophysin expression in the CA3region in hippocampus of vascular dementia (VD) rats.Methods The

  14. Basic fibroblast growth factor-loaded, mineralized biopolymer-nanofiber scaffold improves adhesion and proliferation of rat mesenchymal stem cells.

    Science.gov (United States)

    Kim, Tae-Hyun; Kim, Jung-Ju; Kim, Hae-Won

    2014-02-01

    Nanofibrous matrices are attractive scaffolding platforms for tissue regeneration. Modification of the nanofiber surface, particularly with biological proteins, improves cellular interactions. Here, we loaded basic fibroblast growth factor (bFGF) onto mineralized nanofibers and investigated the effect on adhesion and proliferation of rat mesenchymal stem cells. bFGF loading was significantly higher on the mineralized nanofiber than on the non-mineralized one. Release of bFGF from the mineralized nanofibers was continuous over 2 weeks. Cells cultured on the bFGF-loaded nanofiber attached and proliferated in significantly higher numbers than those on the bFGF-free nanofiber. bFGF-receptor inhibition study confirmed the biological role played by the loaded bFGF. This study details the advantages of the mineralized nanofiber surface for the loading and delivery bFGF, and thus the bFGF-loaded nanofiber scaffold may be useful for tissue repair and regeneration.

  15. Collagen-GAG Scaffolds Grafted Onto Myocardial Infarcts in a Rat Model: A Delivery Vehicle for Mesenchymal Stem Cells

    Institute of Scientific and Technical Information of China (English)

    Z. XIANG; R. LIAO; M. KELLY; M. SPECTOR

    2005-01-01

    @@ 1 Introduction The objective of the present study was to investigate the response of rat myocardial scar tissue to type Ⅰ collagen-glycosaminoglycan (GAG) tissue engineering scaffolds, and to assess the feasibility of using a collagenGAG scaffold as a delivery vehicle for bone marrow-derived mesenchymal stem cells. The benefits of employing the collagen-GAG scaffold for this application include the following: (1) the large surface area of the three-dimensional sponge-like material allows for the delivery of a high cell density to the infarct site; (2) the scaffold allows for the localization and retention of the cells at the site of implantation; (3) the tissue response to the scaffold may promote angiogenesis at the implant site.

  16. The mobilization of rat's mesenchymal stem cells into peripheral blood by LiCL and its potency differentiation

    Institute of Scientific and Technical Information of China (English)

    DENG Jun; ZOU ZhongMin; ZHOU TaoLi; AI GuoPing; WANG JunPing; DONG ShiWu; SU YongPing

    2008-01-01

    Mesenchymal stem cells (MSCs) are multipotent cells, which can differentiate into different tissues. It is still controversial whether MSCs can be isolated from adult peripheral blood (PB) under normal condi-tions and whether they can be mobilized in a way similar to that of hematopoietic stem cells (HSCs), In this study, rat MSCs circulating in the PB under normal conditions can be isolated and cultured, and MSCs from PB and BM can be mobilized by LiCL. The mobilized MSCs can be induced to differentiate into neuron by such factors as β-mercaptoethanol and supernatants of nerve cell cultures. The present study provides a broader perspective on the repair of neural trauma.

  17. In vitro Culture of Bone Marrow Mesenchymal Stem Cells in Rats and Differentiation into Retinal Neural-like Cells

    Institute of Scientific and Technical Information of China (English)

    SUN Xufang; JIANG Huanrong; YANG Hong

    2007-01-01

    In order to study the in vitro culture and expansion of bone marrow mesenchymal stem cells in rats (rMSCs) and the possibility of rMSCs differentiation into retinal neural cells, the bone marrow-derived cells in SD rats were isolated and cultured in vitro. The retinal neural cells in SD rats were cultured and the supernatants were collected to prepare conditioned medium. The cultured rMSCs were induced to differentiate by two steps. Imrnunofluorescence method and anti-nestin, anti-NeuN, anti-GFAP and anti-Thy1.1 antibodies were used to identify the cells derived from the rMSCs. The results showed that the in vitro cultured rMSCs grew well and expanded quickly. After induction with two conditioned media, rMSCs was induced to differentiate into neural progenitor cells, then into retinal neural-like cells which were positive for nestin, NeuN, GFAP and Thy1.1 de-tected by fluorescence method. The findings suggested that rMSCs could be culture and expanded in vitro, and induced to differentiate into retinal neural-like cells.

  18. In vitro cultivation of rat bone marrow mesenchymal stem cells and establishment of pEGFP/Ang-1 transfection method

    Institute of Scientific and Technical Information of China (English)

    Xiu-Qun Zhang; Long Wang; Shu-Li Zhao; Wei Xu

    2014-01-01

    Objective:To obtain the bone marrow mesenchymal stem cells (BMSCs), complete phenotypic identification and successfully transfect rat BMSCs by recombinant plasmid pEGFP/Ang-1. Methods:BMSCs were isolated from bone marrow using density gradient centrifugation method and adherence screening method, and purified. Then the recombinant plasmid pEGFP/Ang-1 was used to transfect BMSCs and the positive clones were obtained by the screen of G418 and observed under light microscopy inversely. Green fluorescent exhibited by protein was enhanced to measure the change time of the expression amount of Ang-1. Results: BMSCs cell lines were obtained successfully by adherence screening method and density gradient centrifugation. Ang-1 recombinant plasmid was transfected smoothly into rat BMSCs, which can express Ang-1 for 3 d and decreased after 7 d. Conclusions:Adherence screening method and density gradient centrifugation can be effective methods to obtain BMSCs with high purity and rapid proliferation. Besides, the expression of transfected recombinant plasmid pEGFP/Ang-1 in rat BMSCs is satisfactory.

  19. Transplantation of bone marrow mesenchymal stem cells on collagen scaffolds for the functional regeneration of injured rat uterus.

    Science.gov (United States)

    Ding, Lijun; Li, Xin'an; Sun, Haixiang; Su, Jing; Lin, Nacheng; Péault, Bruno; Song, Tianran; Yang, Jun; Dai, Jianwu; Hu, Yali

    2014-06-01

    Serious injuries of endometrium in women of reproductive age are often followed by uterine scar formation and a lack of functional endometrium predisposing to infertility or miscarriage. Bone marrow-derived mesenchymal stem cells (BM-MSCs) have shown great promise in clinical applications. In the present study, BM-MSCs loaded onto degradable collagen membranes were constructed. Collagen membranes provided 3-dimmensional architecture for the attachment, growth and migration of rat BM-MSCs and did not impair the expression of the stemness genes. We then investigated the effect of collagen/BM-MSCs constructs in the healing of severe uterine injury in rats (partial full thickness uterine excision). At four weeks after the transplantation of collagen/BM-MSCs constructs, BM-MSCs were mainly located to the basal membrane of regenerative endometrium. The wounded tissue adjacent to collagen/BM-MSCs constructs expressed higher level of bFGF, IGF-1, TGFβ1 and VEGF than the corresponding tissue in rats receiving collagen construct alone or in spontaneous regeneration group. Moreover, the collagen/BM-MSCs system increased proliferative abilities of uterine endometrial and muscular cells, facilitated microvasculature regeneration, and restored the ability of endometrium to receive the embryo and support its development to a viable stage. Our findings indicate that BM-MSCs may support uterine tissue regeneration.

  20. Nampt Expression Decreases Age-Related Senescence in Rat Bone Marrow Mesenchymal Stem Cells by Targeting Sirt1

    Science.gov (United States)

    Ma, Cao; Pi, Chenchen; Yang, Yue; Lin, Lin; Shi, Yingai; Li, Yan; Li, Yulin; He, Xu

    2017-01-01

    Senescence restricts the development of applications involving mesenchymal stem cells (MSCs) in research fields, such as tissue engineering, and stem cell therapeutic strategies. Understanding the mechanisms underlying natural aging processes may contribute to the development of novel approaches to preventing age-related diseases or slowing individual aging processes. Nampt is a rate-limiting NAD biosynthetic enzyme that plays critical roles in energy metabolism, cell senescence and maintaining life spans. However, it remains unknown whether Nampt influences stem cell senescence. In this study, the function of Nampt was investigated using a rat model of natural aging. Our data show that Nampt expression was significantly lower in MSCs obtained from aged rats than in those obtained from young rats during physiological aging. Reducing the level of Nampt in aged MSCs resulted in lower intracellular concentrations of NAD+ and downregulated Sirt1 expression and activity. After the Nampt inhibitor FK866 was added, young MSCs were induced to become aged cells. The enhanced senescence was correlated with NAD+ depletion and Sirt1 activity attenuation. In addition, Nampt overexpression attenuated cell senescence in aged MSCs. Our findings provide a new explanation for the mechanisms underlying stem cell senescence and a novel target for delaying stem cell senescence and preventing and treating age-related diseases. PMID:28125705

  1. Intravenous Injections of Human Mesenchymal Stromal Cells Modulated the Redox State in a Rat Model of Radiation Myelopathy

    Directory of Open Access Journals (Sweden)

    Jing Zhang

    2015-01-01

    Full Text Available The main aim of the present study was to assess the antioxidative effects of human umbilical cord-derived mesenchymal stromal cells (UC-MSCs in a rat model of radiation myelopathy. UC-MSCs were isolated from Wharton’s jelly (WJ of umbilical cords. An irradiated cervical spinal cord rat model (C2-T2 segment was generated using a 60Co irradiator to deliver 30 Gy of radiation. UC-MSCs were injected through the tail vein at 90 days, 97 days, 104 days, and 111 days after-irradiation. Histological damage was examined by cresyl violet/Nissl staining. The activities of two antioxidant enzymes catalase (CAT and glutathione peroxidase (GPX in the spinal cord were measured by the biomedical assay. In addition, the levels of vascular endothelial growth factor (VEGF and angiopoietin-2 (Ang-2 in the spinal cord were determined by ELISA methods. Multiple injections of UC-MSCs through the tail vein ameliorated neuronal damage in the spinal cord, increased the activities of the antioxidant enzymes CAT and GPX, and increased the levels of VEGF and Ang-2 in the spinal cord. Our results suggest that multiple injections of UC-MSCs via the tail vein in the rat model of radiation myelopathy could significantly improve the antioxidative microenvironment in vivo.

  2. Repair of Rat Segmental Defects with Mineralized Collagen Grafts Combined with or without Mesenchymal Stem Cells and BMP-2

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    The aim of the present study was to investigate and compare the bone formation capacity with three different grafts. Four millimeter segmental defects were created in adult rat tibias and were either left empty(control defects) or implanted with (1) nano-hydroxyapatite/collagen/PLA (nHAC/PLA) composite, (2)nHAC/ PLA composite added with bone marrow mesenchymal stem cells ( BMSCs ), (3) nHAC/PLA composite added with bone morphogenetic protein 2 ( BMP- 2). Radiographs of the defects were taken weekly post-surgery.After 1 or 2 months, the rats were euthanized. Histologic analyses were performed on the harvested tissue.nHAC/ PLA composite could enhance the repair of rat tibia segmental defects. Addition of BMSCs or BMP- 2 to nHAC/ PLA led to an increase in osteogenesis. nHAC/ PLA composite could be an ideal alternative bone-graft material and it could also be used as an ideal carrier of BMSCs or BMP- 2.

  3. Chemically Induced Breast Tumors in Rats Are Detectable in Early Stages by Contrast Enhanced Magnetic Resonance Imaging but Not by Changes in the Acute-Phase Reactants in Serum

    Directory of Open Access Journals (Sweden)

    Onn Haji Hashim

    2011-02-01

    Full Text Available The present study was undertaken to develop a rat model for monitoring the early development of breast cancer. Twelve female rats were divided into two groups of six rats that were either treated with N-methyl-N-nitrosourea to induce breast cancer or with bacterial lipopolysaccharide to induce inflammation. Serum samples taken from the rats prior to the treatment were used as controls. By the 14th week, presence of the tumor was detectable by contrast enhanced magnetic resonance imaging and confirmed by histopathology. When the serum proteins of the rats were examined by 2-dimensional electrophoresis (2-DE, no difference could be detected in the profiles of all proteins before and 18 weeks after administration of N-methyl-N-nitrosourea. However, higher expression of alpha-1B glycoprotein was detectable by 2-DE in serum samples of rats at the 18th week post-treatment with lipopolysaccharide.

  4. Effects of Nitric Oxide Production Inhibitor Named, NG-Nitro-L-Arginine Methyl Ester (L-NAME, on Rat Mesenchymal Stem Cells Differentiation

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    E Arfaei

    2010-04-01

    Full Text Available Introduction & Objectives: Recently, the findings of some studies have shown that, nitric oxide (NO probably has an important role in differentiation of mesenchymal stem cells to osteoblasts. The aim of the present investigation was to study the effects of nitric oxide production inhibitor named, NG-nitro-L-arginine methyl ester (L-NAME, on rat mesenchymal stem cells differentiation to osteoblasts in vitro. Materials & Methods: This was an experimental study conducted at Hamedan University of Medical Sciences in 2009, in which rat bone marrow stem cells were isolated in an aseptic condition and cultured in vitro. After third passage, the cells were cultured in osteogenic differentiation medium. To study the effects of L-NAME on osteogenic differentiation, the L-NAME was added to the culture medium at a concentration of 125, 250, and 500 μM in some culture plates. During the culture procedure, the media were replaced with fresh ones, with a three days interval. After 28 days of culturing the mineralized matrix was stained using Alizarian red staining method. The gathered data were analyzed by SPSS software version 12 using one way ANOVA. Results: The findings of this study showed that in the presence of L-NAME, differentiation of bone marrow mesenchymal stem cells to osteoblasts was disordered and matrix mineralization significantly decreased in a dose dependent manner. Conclusion: This study revealed that, inhibition of nitric oxide production using L-NAME can prevent the differentiation of rat bone marrow mesenchymal stem cells to osteoblast. The results imply that NO is an important constituent in differentiation of mesenchymal stem cell to osteoblasts.

  5. The preventive effects of neural stem cells and mesenchymal stem cells intra-ventricular injection on brain stroke in rats

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    Seyed Mojtaba Hosseini

    2015-01-01

    Full Text Available Introduction: Stroke is one of the most important causes of disability in developed countries and, unfortunately, there is no effective treatment for this major problem of central nervous system (CNS; cell therapy may be helpful to recover this disease. In some conditions such as cardiac surgeries and neurosurgeries, there are some possibilities of happening brain stroke. Inflammation of CNS plays an important role in stroke pathogenesis, in addition, apoptosis and neural death could be the other reasons of poor neurological out come after stroke. In this study, we examined the preventive effects of the neural stem cells (NSCs and mesenchymal stem cells (MSCs intra-ventricular injected on stroke in rats. Aim: The aim of this study was to investigate the preventive effects of neural and MSCs for stroke in rats. Materials and Methods: The MSCs were isolated by flashing the femurs and tibias of the male rats with appropriate media. The NSCs were isolated from rat embryo ganglion eminence and they cultured NSCs media till the neurospheres formed. Both NSCs and MSCs were labeled with PKH26-GL. One day before stroke, the cells were injected into lateral ventricle stereotactically. Results: During following for 28 days, the neurological scores indicated that there are better recoveries in the groups received stem cells and they had less lesion volume in their brain measured by hematoxylin and eosin staining. Furthermore, the activities of caspase-3 were lower in the stem cell received groups than control group and the florescent microscopy images showed that the stem cells migrated to various zones of the brains. Conclusion: Both NSCs and MSCs are capable of protecting the CNS against ischemia and they may be good ways to prevent brain stroke consequences situations.

  6. Erythropoietin Modification Enhances the Protection of Mesenchymal Stem Cells on Diabetic Rat-Derived Schwann Cells: Implications for Diabetic Neuropathy

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    Shuyun Zhang

    2017-01-01

    Full Text Available Diabetes-triggered apoptosis of Schwann cells (SC contributes to the degradation of diabetic peripheral neuropathy (DNP. In recent years, mesenchymal stem cells (MSC were applied to DPN repair and it was demonstrated that paracrine secretion played a key role in neuroprotection exerted by MSC. Erythropoietin (EPO is a potent cytokine capable of reducing apoptosis of SC. However, the expression of EPO in MSC is limited. In this study, we hypothesized that overexpression of EPO in MSC (EPO-MSC may significantly improve their neuroprotective potentials. The EPO overexpression in MSC was achieved by lentivirus transduction. SC derived from the periphery nerve of diabetic rats were cocultured with MSC or EPO-MSC in normal or high glucose culture condition, respectively. In normal glucose culture condition, the overexpression of EPO in MSC promoted the MSC-induced restoration of SC from diabetic rats, including increases in GSH level and cell viability, decrease in TUNEL apoptosis, upregulation of antiapoptotic proteins, p-Akt, and Bcl-2, and downregulation of proapoptotic proteins, cleaved caspase-3, and Bax. The subsequent results in high glucose culture condition showed similar promotions achieved by EPO-MSC. Thus, it could be concluded that EPO-MSC possessed a potent potential in hampering apoptosis of SC, and the suppression was probably attributed to attenuating oxidative stress and regulating apoptosis related protein factors.

  7. Neuroprotective Effects of Bone Marrow Mesenchymal Stem Cells on Bilateral Common Carotid Arteries Occlusion Model of Cerebral Ischemia in Rat

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    Bagher Pourheydar

    2016-01-01

    Full Text Available Cell therapy is the most advanced treatment of the cerebral ischemia, nowadays. Herein, we discuss the neuroprotective effects of bone marrow mesenchymal stem cells (BMSCs on rat hippocampal cells following intravenous injection of these cells in an ischemia-reperfusion model. Adult male Wistar rats were divided into 5 groups: control, sham (surgery without blockage of common carotid arteries, ischemia (common carotid arteries were blocked for 30 min prior to reperfusion, vehicle (7 days after ischemia PBS was injected via the tail vein, and treatment (injections of BMSC into the tail veins 7 days after ischemia. We performed neuromuscular and vestibulomotor function tests to assess behavioral function and, finally, brains were subjected to hematoxylin and eosin (H&E, anti-Brdu immunohistochemistry, and TUNEL staining. The ischemia group had severe apoptosis. The group treated with BMSCs had a lower mortality rate and also had significant improvement in functional recovery (P<0.001. Ischemia-reperfusion for 30 min causes damage and extensive neuronal death in the hippocampus, especially in CA1 and CA3 regions, leading to several functional and neurological deficits. In conclusion, intravenous injection of BMSCs can significantly decrease the number of apoptotic neurons and significantly improve functional recovery, which may be a beneficial treatment method for ischemic injuries.

  8. Application of Bone Marrow-Derived Mesenchymal Stem Cells in the Treatment of Intrauterine Adhesions in Rats

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    Jianmei Wang

    2016-09-01

    Full Text Available Aims: To investigate the therapeutic effects of bone marrow-derived mesenchymal stem cells (BMSCs transplantation on intrauterine adhesions (IUA. Methods: BMSCs were isolated and labeled by green fluorescence protein. IUA model was established by mechanical injury. 48 rats were randomly divided into control, IUA model, BMSCs vein injection and BMSCs intrauterine injection groups (n=12 in each group. The third generation of BMSCs was injected through tail vein or intrauterine. Three rats were killed at time 0 h, 7 d, 14 d and 28 d and bilateral uterus were obtained at each time points for the subseqent experiments. Morphological changes were determined by hematoxylin-eosin staining or Masson staining. Estrogen receptor (ER and progesterone receptor (PR were detected by immunohistochemistry. Results: BMSCs were specifically stained by CD44 and CD90, but not by CD45. Before treatment, the numbers of endometrial glands were significantly decreased, while fibrosis area rate was increased in IUA model group (PConclusion: BMSCs transplantation was effective to repair the damaged endometrium likely through promoting the ER and PR expressions.

  9. Effects of Exendine-4 on The Differentiation of Insulin Producing Cells from Rat Adipose-Derived Mesenchymal Stem Cells

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    Layasadat Khorsandi

    2016-02-01

    Full Text Available Objective: To evaluate the effect of Exendine-4 (EX-4, a Glucagon-like peptide 1 (GLP-1 receptor agonist, on the differentiation of insulin-secreting cells (IPCs from rat adipose-derived mesenchymal stem cells(ADMSCs. Materials and Methods: In this experimental study, ADMSCs were isolated from rat adipose tissue and exposed to induction media with or without EX-4. After induction, the existence of IPCs was confirmed by morphology analysis, expression pattern analysis of islet-specific genes (Pdx-1, Glut-2 and Insulin and insulin synthesis and secretion. Results: IPCs induced in presence of EX-4 were morphologically similar to pancreatic islet-like cells. Expression of Pdx-1, Glut-2 and Insulin genes in EX-4 treated cells was significantly higher than the cells exposed to differentiation media without EX-4. Compared to EX-4 untreated ADMSCs, insulin release from EX-4 treated ADMSCs showed a nearly 2.5 fold (P<0.05 increase when exposed to a high glucose (25 mM medium. The percentage of insulin positive cells in the EX-4 treated group was approximately 4-fold higher than in the EX-4 untreated ADMSCs. Conclusion: The present study has demonstrated that EX-4 enhances the differentiation of ADMSCs into IPCs. Improvement of this method may help the formation of an unlimited source of cells for transplantation.

  10. Adenovirus-Mediated Over-Expression of Nrf2 Within Mesenchymal Stem Cells (MSCs Protected Rats Against Acute Kidney Injury

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    Mohammad Mohammadzadeh-Vardin

    2015-06-01

    Full Text Available Purpose: Recent developments in the field of cell therapy have led to a renewed interest in treatment of acute kidney injury (AKI. However, the early death of transplanted mesenchymal stem cells (MSCs in stressful microenvironment of a recipient tissue is a major problem with this kind of treatment. The objective of this study was to determine whether overexpression of a cytoprotective factor, nuclear factor erythroid-2 related factor 2 (Nrf2, in MSCs could protect rats against AKI. Methods: The Nrf2 was overexpressed in MSCs by recombinant adenoviruses, and the MSCs were implanted to rats suffering from cisplatin-induced AKI. Results: The obtained results showed that transplantation with the engineered MSCs ameliorates cisplatin-induced AKI. Morphologic features of the investigated kidneys showed that transplantation with the MSCs in which Nrf2 had been overexpressed significantly improved the complications of AKI. Conclusion: These findings suggested that the engineered MSCs might be a good candidate to be further evaluated in clinical trials. However, detailed studies must be performed to investigate the possible carcinogenic effect of Nrf2 overexpression.

  11. Mesenchymal stem cell-conditioned medium triggers neuroinflammation and reactive species generation in organotypic cultures of rat hippocampus.

    Science.gov (United States)

    Horn, Ana Paula; Bernardi, Andressa; Luiz Frozza, Rudimar; Grudzinski, Patrícia Bencke; Hoppe, Juliana Bender; de Souza, Luiz Fernando; Chagastelles, Pedro; de Souza Wyse, Angela Terezinha; Bernard, Elena Aida; Battastini, Ana Maria Oliveira; Campos, Maria Martha; Lenz, Guido; Nardi, Nance Beyer; Salbego, Christianne

    2011-07-01

    Cell therapy using bone marrow-derived mesenchymal stem cells (MSCs) seems to be a new alternative for the treatment of neurodegenerative diseases. Despite several promising results with their use, possible side effects are still unknown. In a previous work, we have shown that MSC-conditioned medium is toxic to hippocampal slice cultures and aggravates cell death induced by oxygen and glucose deprivation. In this work, we investigated whether the inflammatory response and/or reactive species formation could be involved in that toxicity. Rat organotypic hippocampal cultures were exposed for 24 h to conditioned medium from MSCs isolated from rat bone marrow. A marked glial activation was observed after exposure of cultures to MSC-conditioned medium, as evidenced by glial fibrillary acid protein (GFAP) and isolectin B(4) increase. Tumor necrosis factor-α and interleukin-6 levels were increased in the culture medium, and 2,7-dihydrodichlorofluorescein diacetate oxidation (indicating reactive species generation) and inducible nitric oxide synthase (iNOS) immunocontent were also higher after exposure of cultures to MSC-conditioned medium. Antioxidants (ascorbic acid and TROLOX(®)), N(ω)-nitro-l-arginine methyl ester hydrochloride, and anti-inflammatory drugs (indomethacin and dexamethasone) reduced cell death in hippocampal organotypic cultures after their exposure to MSC-conditioned medium. The results obtained here suggest that MSC-secreted factors trigger reactive species generation and neuroinflammation in organotypic cultures of hippocampus, introducing a note of caution in the use of these cells for neurological application.

  12. Dynamic Tracking of Injected Mesenchymal Stem Cells after Myocardial Infarction in Rats: A Serial 7T MRI Study

    Science.gov (United States)

    Chen, Xiuyu; Lu, Minjie; Ma, Ning; Yin, Gang; Cui, Chen

    2016-01-01

    Purpose. To track the fate of micron-sized particles of iron oxide (MPIO) labeled mesenchymal stem cells (MSCs) in vivo in a rat myocardial infarction model using 7T magnetic resonance imaging (MRI) scanner. Materials and Methods. Male MSCs (2 × 106/50 μL) dual-labeled with MPIO and CM-DiI were injected into the infarct periphery 7 days after myocardial infarction (MI). The control group received cell-free media injection. The temporal stem cell location, signal intensity, and cardiac function were dynamically assessed using a 7T MRI at 24 h before transplantation (baseline), 3 days, 2 weeks, and 4 weeks after transplantation, respectively. Results. MR hypointensities caused by MPIOs were observed on T2⁎-weighted images at all time points after MSCs injection. Cine-MRI showed that MSCs moderated progressive left ventricular remodeling. Double staining for iron and CD68 revealed that most of the iron-positive cells were CD68-positive macrophages. Real-time PCR for rat SRY gene showed the number of survival MSCs considerably decreased after transplantation. MSC-treated hearts had significantly increased capillary density in peri-infarct region and lower cardiomyocytes apoptosis and fibrosis formation. Conclusions. Iron particles are not a reliable marker for in vivo tracking the long-term fate of MSCs engraftment. Despite of poor cell retention, MSCs moderate left ventricular remodeling after MI. PMID:27656215

  13. Erythropoietin Modification Enhances the Protection of Mesenchymal Stem Cells on Diabetic Rat-Derived Schwann Cells: Implications for Diabetic Neuropathy

    Science.gov (United States)

    Zhang, Shuyun

    2017-01-01

    Diabetes-triggered apoptosis of Schwann cells (SC) contributes to the degradation of diabetic peripheral neuropathy (DNP). In recent years, mesenchymal stem cells (MSC) were applied to DPN repair and it was demonstrated that paracrine secretion played a key role in neuroprotection exerted by MSC. Erythropoietin (EPO) is a potent cytokine capable of reducing apoptosis of SC. However, the expression of EPO in MSC is limited. In this study, we hypothesized that overexpression of EPO in MSC (EPO-MSC) may significantly improve their neuroprotective potentials. The EPO overexpression in MSC was achieved by lentivirus transduction. SC derived from the periphery nerve of diabetic rats were cocultured with MSC or EPO-MSC in normal or high glucose culture condition, respectively. In normal glucose culture condition, the overexpression of EPO in MSC promoted the MSC-induced restoration of SC from diabetic rats, including increases in GSH level and cell viability, decrease in TUNEL apoptosis, upregulation of antiapoptotic proteins, p-Akt, and Bcl-2, and downregulation of proapoptotic proteins, cleaved caspase-3, and Bax. The subsequent results in high glucose culture condition showed similar promotions achieved by EPO-MSC. Thus, it could be concluded that EPO-MSC possessed a potent potential in hampering apoptosis of SC, and the suppression was probably attributed to attenuating oxidative stress and regulating apoptosis related protein factors.

  14. Proliferation and Differentiation of Rat Osteoporosis Mesenchymal Stem Cells (MSCs) after Telomerase Reverse Transcriptase (TERT) Transfection

    OpenAIRE

    Li, Chao; Wei, Guojun; Gu, Qun; Wang, Qiang; Tao, Shuqin; Liang XU

    2015-01-01

    Background The aim of this study was to determine whether MSC are excellent materials for MSCs transplantation in the treatment of osteoporosis. Material/Methods We studied normal, osteoporosis, and TERT-transfected MSC from normal and osteoporosis rats to compare the proliferation and osteogenic differentiation using RT-PCR and Western blot by constructing an ovariectomized rat model of osteoporosis (OVX). The primary MSC from model rats were extracted and cultured to evaluate the proliferat...

  15. Microbubble-mediated ultrasound promotes accumulation of bone marrow mesenchymal stem cell to the prostate for treating chronic bacterial prostatitis in rats

    OpenAIRE

    2016-01-01

    Chronic bacterial prostatitis (CBP) is an intractable disease. Although bone marrow mesenchymal stem cells (BMMSCs) are able to regulate inflammation in CBP, the effect of microbubble-mediated ultrasound- induced accumulation of BMMSCs on CBP remains unclear. To address this gap, a model of CBP was established in SD rats, which were then treated with BMMSCs alone (BMMSC group), BMMSCs with ultrasound (ultrasound group), BMMSCs with microbubble-mediated ultrasound (MMUS group) and compared wit...

  16. Immunomodulatory effects of bone marrow mesenchymal stem cells derived from homologous recipients in rats after heart transplantation

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    De-zhong LIU

    2012-03-01

    Full Text Available Objective To observe the immunomodulatory effects of homologous bone marrow mesenchymal stem cells (MSCs obtained from the bone marrow in rats after heart transplantation. Methods Twenty adult male Lewis rats were used as donors for the heart transplantation, whereas twenty adult male Wistar rats served as recipients. The recipients with cervical heart transplantation were randomly divided into two groups (10 each. Approximately 3ml 0.9% NaCl solution was injected through the tail vein 24h after heart transplantation in the control group (group A. About 2×106 MSCs (suspended in 3ml 0.9% NaCl solution were injected through the tail vein 24h after heart transplantation in the MSCs treatment group (group B. Four recipient rats from each group were randomly chosen one week after transplantation for determining proportion of CD4+ T, CD8+ T, CD4+CD25high T, and CD4+CD25highfoxp3+ T cells in the lymphocytes in the venous blood and grafts. Subsequently, the CD4+/CD8+ ratio was calculated. The survival time of the grafts were observed in the remaining six rats in each group. Results (1The survival time of the transplanted hearts was 7.2±1.3d in group A, and 14.8±2.9d in group B, showing a significant difference between the two groups (P 0.05. The ratios of CD4+CD25high T cells/total lymphocytes and CD4+CD25highFoxp3+ T cells/ total lymphocytes in the allografts were evidently higher in group B (2.74%±0.28%, 2.54%±0.31% than in group A (0.61%±0.06%, 0.53%±0.06%, showing a significant statistical difference (P < 0.01. Conclusion Intravenous infusion with MSCs from the bone marrow of the recipients can induce immune tolerance and prolong the survival time of transplanted heart in rats.

  17. Cryopreservation of Rat Bone Marrow Derived Mesenchymal Stem Cells by Two Conventional and Open-pulled Straw Vitrification Methods

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    Mohammad Hadi Bahadori

    2009-01-01

    Full Text Available Objective: Mesenchymal stem cells (MSCs are obtained from a variety of sources, mainlythe bone marrow. These cells have a great potential for clinical research, however they cannotstay alive for long periods in culture. The aim of this study is to determine whether vitrificationcan be a useful freezing method for the storage of MSCs.Materials and Methods: Mesenchymal stem cells were isolated from rat bone marrow basedon their capacity to adhere to plastic culture surfaces. MSCs were cryopreserved using boththe vitrification method and open-pulled straw (OPS vitrification and stored in liquid nitrogenwith ethylene glycol ficoll (EFS as a cryoprotectant for two months. The morphology andviability of thawed MSCs were evaluated by trypan blue staining. Furthermore, pre and postcryopreserved MSCs were induced to osteocyte and adipocyte with corresponding osteogenicand adipogenic medium.Results: After thawing, the viability rates were 81.33% ± 6.83 for the vitrification method and80.83% ± 6.4 for OPS vitrification, while the values in the pre-vitrification control group were88.16% ± 6.3 (Mean ± SD, n = 6. Post-cryopreserved cells from both the vitrification methodand OPS vitrification also had a similar cellular morphology and colony-formation that wasindistinguishable from non-vitrified fresh MSCs. In addition, the resuscitated cells cultured ininduction medium showed osteogenesis. Mineral production and deposition was detectableby alizarine red S staining. Moreover, by applying an adipogenic differentiation condition,both pre and post cryopreserved cells differentiated into adipocyte and lipid vacuole accumulationthat was stained by oil red O.Conclusion: Vitrification is a reliable and effective method for the cryopreservation of MSCs.

  18. Propofol injection combined with bone marrow mesenchymal stem cell transplantation better improves electrophysiological function in the hindlimb of rats with spinal cord injury than monotherapy

    Institute of Scientific and Technical Information of China (English)

    Yue-xin Wang; Jing-jing Sun; Mei Zhang; Xiao-hua Hou; Jun Hong; Ya-jing Zhou; Zhi-yong Zhang

    2015-01-01

    The repair effects of bone marrow mesenchymal stem cell transplantation on nervous system damage are not satisfactory. Propofol has been shown to protect against spinal cord injury. Therefore, this study sought to explore the therapeutic effects of their combination on spinal cord injury. Rat models of spinal cord injury were established using the weight drop method. Rats were subjected to bone marrow mesenchymal stem cell transplantationvia tail vein injection and/or propofol injectionvia tail vein using an infusion pump. Four weeks after cell transplan-tation and/or propofol treatment, the cavity within the spinal cord was reduced. The numbers of PKH-26-positive cells and horseradish peroxidase-positive nerve ifbers apparently increased in the spinal cord. Latencies of somatosensory evoked potentials and motor evoked potentials in the hindlimb were noticeably shortened, amplitude was increased and hindlimb motor function was obviously improved. Moreover, the combined effects were better than cell transplantation or propofol injection alone. The above data suggest that the combination of propofol injection and bone marrow mesenchymal stem cell transplantation can effectively improve hindlimb electro-physiological function, promote the recovery of motor funtion, and play a neuroprotective role in spinal cord injury in rats.

  19. Propofol combined with bone marrow mesenchymal stem cell transplantation improves electrophysiological function in the hindlimb of rats with spinal cord injury better than monotherapy

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    Yue-xin Wang

    2015-01-01

    Full Text Available The repair effects of bone marrow mesenchymal stem cell transplantation on nervous system damage are not satisfactory. Propofol has been shown to protect against spinal cord injury. Therefore, this study sought to explore the therapeutic effects of their combination on spinal cord injury. Rat models of spinal cord injury were established using the weight drop method. Rats were subjected to bone marrow mesenchymal stem cell transplantation via tail vein injection and/or propofol injection via tail vein using an infusion pump. Four weeks after cell transplantation and/or propofol treatment, the cavity within the spinal cord was reduced. The numbers of PKH-26-positive cells and horseradish peroxidase-positive nerve fibers apparently increased in the spinal cord. Latencies of somatosensory evoked potentials and motor evoked potentials in the hindlimb were noticeably shortened, amplitude was increased and hindlimb motor function was obviously improved. Moreover, the combined effects were better than cell transplantation or propofol injection alone. The above data suggest that the combination of propofol injection and bone marrow mesenchymal stem cell transplantation can effectively improve hindlimb electrophysiological function, promote the recovery of motor funtion, and play a neuroprotective role in spinal cord injury in rats.

  20. Mesenchymal stem cells improve the outcomes of liver recipients via regulating CD4+ T helper cytokines in rats

    Institute of Scientific and Technical Information of China (English)

    Yang Yang; Zhong-Yang Shen; Bin Wu; Ming-Li Yin; Bo-Ya Zhang; Hong-Li Song

    2015-01-01

    BACKGROUND: Bone marrow mesenchymal stem cells (BMMSCs) exert immunosuppressive activities in transplantation. This study aimed to determine whether BMMSCs reduce acute re-jection and improve outcomes of liver transplantation in rats. METHODS: Orthotopic liver transplantation from Lewis to Brown Norway rats was performed, which was followed by the infusion of BMMSCs through the penile superifcial dorsal vein. Normal saline infusion was used as a control. Animals were sacriifced at 0, 24, 72, or 168 hours after BMMSCs infu-sion. Liver grafts, and recipient serum and spleen tissues were obtained. Histopathology, apoptosis, serum liver enzymes, se-rum cytokines, and circulating regulatory T (Treg), Th1, Th2 and Th17 cells were assessed at each time point. RESULTS: BMMSCs signiifcantly attenuated acute rejection and improved the survival rate of allogeneic liver transplanta-tion recipients. Liver enzymes and liver apoptosis were signiif-cantly alleviated. The levels of the Th1/Th2 ratio-associated cytokines such as IL-2 and IFN-γ were signiifcantly reduced and IL-10 was signiifcantly increased. The levels of the Th17/Tregs axis-associated cytokines such as IL-6, IL-17, IL-23, and TNF-α were signiifcantly reduced, whereas TGF-β concentra-tion was signiifcantly increased. Moreover, lfow cytometry analysis showed that the infusion of BMMSCs signiifcantly increased Th2 and Treg cells and decreased Th1 and Th17 cells. CONCLUSION: BMMSCs had immunomodulatory effects, at-tenuated acute rejection and improved outcomes of allogeneic liver transplantation in rats by regulating the levels of cyto-kines associated with Th1/Th2 and Th17/Treg ratios.

  1. Mesenchymal stem cells improve the outcomes of liver recipients via regulating CD4+ T helper cytokines in rats

    Institute of Scientific and Technical Information of China (English)

    Yang Yang; Zhong-Yang Shen; Bin Wu; Ming-Li Yin; Bo-Ya Zhang; Hong-Li Song

    2016-01-01

    BACKGROUND: Bone marrow mesenchymal stem cells (BMMSCs) exert immunosuppressive activities in transplantation. This study aimed to determine whether BMMSCs reduce acute re-jection and improve outcomes of liver transplantation in rats. METHODS: Orthotopic liver transplantation from Lewis to Brown Norway rats was performed, which was followed by the infusion of BMMSCs through the penile superifcial dorsal vein. Normal saline infusion was used as a control. Animals were sacriifced at 0, 24, 72, or 168 hours after BMMSCs infu-sion. Liver grafts, and recipient serum and spleen tissues were obtained. Histopathology, apoptosis, serum liver enzymes, se-rum cytokines, and circulating regulatory T (Treg), Th1, Th2 and Th17 cells were assessed at each time point. RESULTS: BMMSCs signiifcantly attenuated acute rejection and improved the survival rate of allogeneic liver transplanta-tion recipients. Liver enzymes and liver apoptosis were signiif-cantly alleviated. The levels of the Th1/Th2 ratio-associated cytokines such as IL-2 and IFN-γ were signiifcantly reduced and IL-10 was signiifcantly increased. The levels of the Th17/Tregs axis-associated cytokines such as IL-6, IL-17, IL-23, and TNF-α were signiifcantly reduced, whereas TGF-β concentra-tion was signiifcantly increased. Moreover, lfow cytometry analysis showed that the infusion of BMMSCs signiifcantly increased Th2 and Treg cells and decreased Th1 and Th17 cells. CONCLUSION: BMMSCs had immunomodulatory effects, at-tenuated acute rejection and improved outcomes of allogeneic liver transplantation in rats by regulating the levels of cyto-kines associated with Th1/Th2 and Th17/Treg ratios.

  2. Engineered myocardial tissues constructed in vivo using cardiomyocyte-like cells derived from bone marrow mesenchymal stem cells in rats

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    Xing Yujie

    2012-01-01

    Full Text Available Abstract Background To explore the feasibility of constructing engineered myocardial tissues (EMTs in vivo, using polylactic acid -co-glycolic acid (PLGA for scaffold and cardiomyocyte-like cells derived from bone marrow mesenchymal stem cells (BMMSCs for seeded cells. Methods BMMSCs were isolated from femur and tibia of Sprague-Dawley (SD rats by density-gradient centrifugation. The third passage cells were treated with 10 μmol/L 5-azacytidine (5-aza and 0.1 μmol/L angiotensin II (Ang II for 24 h, followed by culturing in complete medium for 3 weeks to differentiated into cardiomyocyte-like cells. The cardiomyocyte-like cells were seeded into PLGA scaffolds to form the grafts. The grafts were cultured in the incubator for three days and then implanted into the peritoneal cavity of SD rats. Four weeks later, routine hematoxylin-eosin (HE staining, immunohistochemical staining for myocardium-specific cardiac troponin I (cTnI, scanning electron microscopy and transmission electron microscopy were used to analyze the morphology and microconstruction of the EMTs in host rats. Results HE staining showed that the cardiomyocyte-like cells distributed equally in the PLGA scaffold, and the nuclei arranged in the spindle shape. Immunohistochemical staining revealed that majority of engrafted cells in the PLGA -Cardiomyocyte-like cells group were positive for cTnI. Scanning electron microscopy showed that the inoculated cells well attached to PLGA and grew in 3 dimensions in construct. Transmission electron microscopy showed that the EMTs contained well arranged myofilaments paralleled to the longitudinal cell axis, the cells were rich in endoplasmic reticulum and mitochondria, while desmosomes, gap junction and Z line-like substances were also can be observed as well within the engrafted cells. Conclusion We have developed an in vivo method to construct engineered myocardial tissue. The in vivo microenvironment helped engrafted cells/tissue survive and

  3. Bone marrow-derived mesenchymal stem cells expressing the Shh transgene promotes functional recovery after spinal cord injury in rats.

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    Jia, Yijia; Wu, Dou; Zhang, Ruiping; Shuang, Weibing; Sun, Jiping; Hao, Haihu; An, Qijun; Liu, Qiang

    2014-06-24

    Spinal cord injury (SCI) is one of the most disabling diseases. Cell-based gene therapy is becoming a major focus for the treatment of SCI. Bone marrow-derived mesenchymal stem cells (BMSCs) are a promising stem cell type useful for repairing SCI. However, the effects of BMSCs transplants are likely limited because of low transplant survival after SCI. Sonic hedgehog (Shh) is a multifunctional growth factor which can facilitate neuronal and BMSCs survival, promote axonal growth, prevent activation of the astrocyte lineage, and enhance the delivery of neurotrophic factors in BMSCs. However, treatment of SCI with Shh alone also has limited effects on recovery, because the protein is cleared quickly. In this study, we investigated the use of BMSCs overexpressing the Shh transgene (Shh-BMSCs) in the treatment of rats with SCI, which could stably secrete Shh and thereby enhance the effects of BMSCs, in an attempt to combine the advantages of Shh and BMSCs and so to promote functional recovery. After Shh-BMSCs treatment of SCI via the subarachnoid, we detected significantly greater damage recovery compared with that seen in rats treated with phosphate-buffered saline (PBS) and BMSCs. Use of Shh-BMSCs increased the expression and secretion of Shh, basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF), improved the behavioral function, enhanced the BMSCs survival, promoted the expression level of neurofilament 200 (NF200), and reduced the expression of glial fibrillary acidic protein (GFAP). Thus, our results indicated that Shh-BMSCs enhanced recovery of neurological function after SCI in rats and could be a potential valuable therapeutic intervention for SCI in humans.

  4. Therapeutic efficacy of differentiated versus undifferentiated mesenchymal stem cells in experimental type I diabetes in rat

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    M.A. Wassef

    2016-03-01

    Full Text Available Selective MSCs differentiation protocol into pancreatic beta cells was conducted in the present study using exendin-4 and TGF-beta. Differentiated and undifferentiated MSCs were assessed in experimental type I diabetes in rats. Ninety female white albino rats were included in the study and divided equally (n=15/group into 6 groups: healthy control, healthy control rats received acellular tissue culture medium, diabetic rats, diabetic rats received acellular tissue culture medium, diabetic rats received undifferentiated MSCs and diabetic rats received differentiated MSCs. Therapeutic efficacy of undifferentiated versus differentiated MSCs was evaluated via assessment of quantitative gene expressions of insulin1, insulin 2, Smad-2, Smad-3, PDX-1, PAX-4, neuroD. Blood glucose and insulin hormone levels were also assessed. Results showed that quantitative gene expressions of all studied genes showed significant decrease in diabetic rat groups. Use of undifferentiated and differentiated MSCs led to a significant elevation of expression levels of all genes with more superior effect with differentiated MSCs except smad-2 gene. As regards insulin hormone levels, use of either undifferentiated or differentiated MSCs led to a significant elevation of its levels with more therapeutic effect with differentiated MSCs. Blood glucose levels were significantly decreased with both undifferentiated and differentiated MSCs in comparison to diabetic groups but its levels were normalized 2 months after injection of differentiated MSCs. In conclusion, use of undifferentiated or differentiated MSCs exhibited significant therapeutic potentials in experimental type I diabetes in rats with more significant therapeutic effect with the use of differentiated MSCs.

  5. Electrophysiological functional recovery in a rat model of spinal cord hemisection injury following bone marrow-derived mesenchymal stem cell transplantation under hypothermia

    Institute of Scientific and Technical Information of China (English)

    Dong Wang; Jianjun Zhang

    2012-01-01

    Following successful establishment of a rat model of spinal cord hemisection injury by resecting right spinal cord tissues, bone marrow stem cells were transplanted into the spinal cord lesions via the caudal vein while maintaining rectal temperature at 34 ± 0.5°C for 6 hours (mild hypothermia). Hematoxylin-eosin staining showed that astrocytes gathered around the injury site and formed scars at 4 weeks post-transplantation. Compared with rats transplanted with bone marrow stem cells under normal temperature, rats transplanted with bone marrow stem cells under hypothermia showed increased numbers of proliferating cells (bromodeoxyuridine-positive cells), better recovery of somatosensory-evoked and motor-evoked potentials, greater Basso, Beattie, and Bresnahan locomotor rating scores, and an increased degree of angle in the incline plate test. These findings suggested that hypothermia combined with bone marrow mesenchymal stem cells transplantation effectively promoted electrical conduction and nerve functional repair in a rat model of spinal cord hemisection injury.

  6. Ex Vivo Gene Therapy Using Human Mesenchymal Stem Cells to Deliver Growth Factors in the Skeletal Muscle of a Familial ALS Rat Model.

    Science.gov (United States)

    Suzuki, Masatoshi; Svendsen, Clive N

    2016-01-01

    Therapeutic protein and molecule delivery to target sites by transplanted human stem cells holds great promise for ex vivo gene therapy. Our group has demonstrated the therapeutic benefits of ex vivo gene therapy targeting the skeletal muscles in a transgenic rat model of familial amyotrophic lateral sclerosis (ALS). We used human mesenchymal stem cells (hMSCs) and genetically modified them to release neuroprotective growth factors such as glial cell line-derived neurotrophic factor (GDNF) and vascular endothelial growth factor (VEGF). Intramuscular growth factor delivery via hMSCs can enhance neuromuscular innervation and motor neuron survival in a rat model of ALS (SOD1(G93A) transgenic rats). Here, we describe the protocol of ex vivo delivery of growth factors via lentiviral vector-mediated genetic modification of hMSCs and hMSC transplantation into the skeletal muscle of a familial ALS rat model.

  7. Use of hybrid chitosan membranes and human mesenchymal stem cells from the Wharton jelly of umbilical cord for promoting nerve regeneration in an axonotmesis rat model

    Institute of Scientific and Technical Information of China (English)

    Andrea G(a)rtner; Yuri Nakamura; Satoshi Hayakawa; Akiyoshi Osakah; Beatriz Porto; Ana Lúcia Luís; Artur SP Varej(a)o; Ana Colette Maurício; Tiago Pereira; Maria Jo(a)o Sim(o)es; Paulo AS Armada-da-Silva; Miguel L Fran(c)a; Rosa Sousa; Simone Bompasso; Stefania Raimondo; Yuki Shirosaki

    2012-01-01

    Many studies have been dedicated to the development of scaffolds for improving post-traumatic nerve regeneration.The goal of this study was to assess the effect on nerve regeneration,associating a hybrid chitosan membrane with non-differentiated human mesenchymal stem cells isolated from Wharton's jelly of umbilical cord,in peripheral nerve reconstruction after crush injury.Chromosome analysis on human mesenchymal stem cell line from Wharton's jelly was carried out and no structural alterations were found in metaphase.Chitosan membranes were previously tested in vitro,to assess their ability in supporting human mesenchymal stem cell survival,expansion,and differentiation.For the in vivo testing,Sasco Sprague adult rats were divided in 4 groups of 6 or 7 animals each:Group 1,sciatic axonotmesis injury without any other intervention (Group 1-Crush); Group 2,the axonotmesis lesion of 3 mm was infiltrated with a suspension of 1 250- 1 500 human mesenchymal stem cells (total volume of 50 μL) (Group 2-CrushCell); Group 3,axonotmesis lesion of 3 mm was enwrapped with a chitosan type Ⅲ membrane covered with a monolayer of non-differentiated human mesenchymal stem cells (Group 3-CrushChitlllCell) and Group 4,axonotmesis lesion of 3 mm was enwrapped with a chitosan type Ⅲ membrane (Group 4-CrushChitlll).Motor and sensory functional recovery was evaluated throughout a healing period of 12 weeks using sciatic functional index,static sciatic index,extensor postural thrust,and withdrawal reflex latency.Stereological analysis was carried out on regenerated nerve fibers.Results showed that infiltration of human mesenchymal stem cells,or the combination of chitosan membrane enwrapment and human mesenchymal stem cell enrichment after nerve crush injury provide a slight advantage to post-traumatic nerve regeneration.Results obtained with chitosan type Ⅲ membrane alone confirmed that they significantly improve post-traumatic axonal regrowth and may represent a very promising

  8. Effect of self-assembled peptide–mesenchymal stem cell complex on the progression of osteoarthritis in a rat model

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    Kim JE

    2014-05-01

    Full Text Available Ji Eun Kim,1 Sang Mok Lee,2 Soo Hyun Kim,1 Phil Tatman,3 Albert O Gee,4 Deok-Ho Kim,3,5 Kyung Eun Lee,6 Youngmee Jung,1 Sang Jun Kim21Biomaterials Research Center, Korea Institute of Science and Technology, Seoul, South Korea; 2Department of Physical and Rehabilitation Medicine, Samsung Medical Center, Seoul, South Korea; 3Department of Bioengineering, 4Department of Orthopaedics and Sports Medicine, 5Institute for Stem Cell and Regenerative Medicine and Center for Cardiovascular Biology, University of Washington, Seattle, WA, USA; 6Advanced Analysis Center, Korea Institute of Science and Technology, Seoul, South KoreaPurpose: To evaluate the efficacy of mesenchymal stem cells (MSCs encapsulated in self-assembled peptide (SAP hydrogels in a rat knee model for the prevention of osteoarthritis (OA progression.Materials and methods: Nanostructured KLD-12 SAPs were used as the injectable hydrogels. Thirty-three Sprague Dawley rats were used for the OA model. Ten rats were used for the evaluation of biotin-tagged SAP disappearance. Twenty-three rats were divided into four groups: MSC (n=6, SAP (n=6, SAP-MSC (n=6, and no treatment (n=5. MSCs, SAPs, and SAP-MSCs were injected into the knee joints 3 weeks postsurgery. Histologic examination, immunofluorescent staining, measurement of cytokine levels, and micro-computed tomography analysis were conducted 6 weeks after injections. Behavioral studies were done to establish baseline measurements before treatment, and repeated 3 and 6 weeks after treatment to measure the efficacy of SAP-MSCs.Results: Concentration of biotinylated SAP at week 1 was not significantly different from those at week 3 and week 6 (P=0.565. Bone mineral density was significantly lower in SAP-MSC groups than controls (P=0.002. Significant differences in terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick-end labeling staining between the control group and all other groups were observed. Caspase-8, tissue inhibitor of

  9. Role of bone marrow-derived mesenchymal stem cells in a rat model of severe acute pancreatitis

    Institute of Scientific and Technical Information of China (English)

    Xiao-Huang Tu; Jing-Xiang Song; Xiao-Jun Xue; Xian-Wei Guo; Yun-Xia Ma; Zhi-Yao Chen; Zhong-Dong Zou; Lie Wang

    2012-01-01

    AIM:To investigate the role and potential mechanisms of bone marrow mesenchymal stem cells (MSCs) in severe acute peritonitis (SAP).METHODS:Pancreatic acinar cells from Sprague Dawley rats were randomly divided into three groups:nonsodium deoxycholate (SDOC) group (non-SODC group),SDOC group,and a MSCs intervention group (i.e.,a co-culture system of MSCs and pancreatic acinar cells + SDOC).The cell survival rate,the concentration of malonaldehyde (MDA),the density of superoxide dismutase (SOD),serum amylase (AMS) secretion rate and lactate dehydrogenase (LDH) leakage rate were detected at various time points.In a separate study,Sprague Dawley rats were randomly divided into either an SAP group or an SAP + MSCs group.Serum AMS,MDA and SOD,interleukin (IL)-6,IL-10,and tumor necrosis factor (TNF)-α levels,intestinal mucosa injury scores and proliferating cells of small intestinal mucosa were measured at various time points after injecting either MSCs or saline into rats.In both studies,the protective effect of MSCs was evaluated.RESULTS:In vitro,The cell survival rate of pancreatic acinar cells and the density of SOD were significantly reduced,and the concentration of MDA,AMS secretion rate and LDH leakage rate were significantly increased in the SDOC group compared with the MSCs intervention group and the Non-SDOC group at each time point.In vivo,Serum AMS,IL-6,TNF-α and MAD level in the SAP + MSCs group were lower than the SAP group;however serum IL-10 level was higher than the SAP group.Serum SOD level was higher than the SAP group at each time point,whereas a significant betweengroup difference in SOD level was only noted after 24 h.Intestinal mucosa injury scores was significantly reduced and the proliferating cells of small intestinal mucosa became obvious after injecting MSCs.CONCLUSION:MSCs can effectively relieve injury to pancreatic acinar cells and small intestinal epithelium,promote the proliferation of enteric epithelium and repair of the mucosa

  10. Transplantation of ATP7B-transduced bone marrow mesenchymal stem cells decreases copper overload in rats.

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    Shenglin Chen

    Full Text Available BACKGROUND: Recent studies have demonstrated that transplantation of ATP7B-transduced hepatocytes ameliorates disease progression in LEC (Long-Evans Cinnamon rats, a model of Wilson's disease (WD. However, the inability of transplanted cells to proliferate in a normal liver hampers long-term treatment. In the current study, we investigated whether transplantation of ATP7B-transduced bone marrow mesenchymal stem cells (BM-MSCs could decrease copper overload in LEC rats. MATERIALS AND METHODS: The livers of LEC rats were preconditioned with radiation (RT and/or ischemia-reperfusion (IRP before portal vein infusion of ATP7B-transduced MSCs (MSCsATP7B. The volumes of MSCsATP7B or saline injected as controls were identical. The expression of ATP7B was analyzed by real-time quantitative polymerase chain reaction (RT-PCR at 4, 12 and 24 weeks post-transplantation. MSCATP7B repopulation, liver copper concentrations, serum ceruloplasmin levels, and alanine transaminase (ALT and aspartate transaminase (AST levels were also analyzed at each time-point post-transplantation. RESULTS: IRP-plus-RT preconditioning was the most effective strategy for enhancing the engraftment and repopulation of transplanted MSCsATP7B. This strategy resulted in higher ATP7B expression and serum ceruloplasmin, and lower copper concentration in this doubly preconditioned group compared with the saline control group, the IRP group, and the RT group at all three time-points post-transplantation (p<0.05 for all. Moreover, 24 weeks post-transplantation, the levels of ALT and AST in the IRP group, the RT group, and the IRP-plus-RT group were all significantly decreased compared to those of the saline group (p<0.05 compared with the IRP group and RT group, p<0.01 compared with IRP-plus-RT group; ALT and AST levels were significantly lower in the IRP-plus-RT group compared to either the IRP group or the RT group (p<0.01 and p<0.05. respectively. CONCLUSIONS: These results demonstrate

  11. Human mesenchymal cells from adipose tissue deposit laminin and promote regeneration of injured spinal cord in rats.

    Science.gov (United States)

    Menezes, Karla; Nascimento, Marcos Assis; Gonçalves, Juliana Pena; Cruz, Aline Silva; Lopes, Daiana Vieira; Curzio, Bianca; Bonamino, Martin; de Menezes, João Ricardo Lacerda; Borojevic, Radovan; Rossi, Maria Isabel Doria; Coelho-Sampaio, Tatiana

    2014-01-01

    Cell therapy is a promising strategy to pursue the unmet need for treatment of spinal cord injury (SCI). Although several studies have shown that adult mesenchymal cells contribute to improve the outcomes of SCI, a description of the pro-regenerative events triggered by these cells is still lacking. Here we investigated the regenerative properties of human adipose tissue derived stromal cells (hADSCs) in a rat model of spinal cord compression. Cells were delivered directly into the spinal parenchyma immediately after injury. Human ADSCs promoted functional recovery, tissue preservation, and axonal regeneration. Analysis of the cord tissue showed an abundant deposition of laminin of human origin at the lesion site and spinal midline; the appearance of cell clusters composed of neural precursors in the areas of laminin deposition, and the appearance of blood vessels with separated basement membranes along the spinal axis. These effects were also observed after injection of hADSCs into non-injured spinal cord. Considering that laminin is a well-known inducer of axonal growth, as well a component of the extracellular matrix associated to neural progenitors, we propose that it can be the paracrine factor mediating the pro-regenerative effects of hADSCs in spinal cord injury.

  12. Prediction of neural differentiation fate of rat mesenchymal stem cells by quantitative morphological analyses using image processing techniques.

    Science.gov (United States)

    Kazemimoghadam, Mahdieh; Janmaleki, Mohsen; Fouani, Mohamad Hassan; Abbasi, Sara

    2015-02-01

    Differentiation of bone marrow mesenchymal stem cells (BMSCs) into neural cells has received significant attention in recent years. However, there is still no practical method to evaluate differentiation process non-invasively and practically. The cellular quality evaluation method is still limited to conventional techniques, which are based on extracting genes or proteins from the cells. These techniques are invasive, costly, time consuming, and should be performed by relevant experts in equipped laboratories. Moreover, they cannot anticipate the future status of cells. Recently, cell morphology has been introduced as a feasible way of monitoring cell behavior because of its relationship with cell proliferation, functions and differentiation. In this study, rat BMSCs were induced to differentiate into neurons. Subsequently, phase contrast images of cells taken at certain intervals were subjected to a series of image processing steps and cell morphology features were calculated. In order to validate the viability of applying image-based approaches for estimating the quality of differentiation process, neural-specific markers were measured experimentally throughout the induction. The strong correlation between quantitative imaging metrics and experimental outcomes revealed the capability of the proposed approach as an auxiliary method of assessing cell behavior during differentiation.

  13. Laminin-521 Promotes Rat Bone Marrow Mesenchymal Stem Cell Sheet Formation on Light-Induced Cell Sheet Technology

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    Zhiwei Jiang

    2017-01-01

    Full Text Available Rat bone marrow mesenchymal stem cell sheets (rBMSC sheets are attractive for cell-based tissue engineering. However, methods of culturing rBMSC sheets are critically limited. In order to obtain intact rBMSC sheets, a light-induced cell sheet method was used in this study. TiO2 nanodot films were coated with (TL or without (TN laminin-521. We investigated the effects of laminin-521 on rBMSCs during cell sheet culturing. The fabricated rBMSC sheets were subsequently assessed to study cell sheet viability, reattachment ability, cell sheet thickness, collagen type I deposition, and multilineage potential. The results showed that laminin-521 could promote the formation of rBMSC sheets with good viability under hyperconfluent conditions. Cell sheet thickness increased from an initial 26.7 ± 1.5 μm (day 5 up to 47.7 ± 3.0 μm (day 10. Moreover, rBMSC sheets maintained their potential of osteogenic, adipogenic, and chondrogenic differentiation. This study provides a new strategy to obtain rBMSC sheets using light-induced cell sheet technology.

  14. Sustained and promoter dependent bone morphogenetic protein expression by rat mesenchymal stem cells after BMP-2 transgene electrotransfer

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    E Ferreira

    2012-07-01

    Full Text Available Transplantation of mesenchymal stem cells (MSCs with electrotransferred bone morphogenetic protein-2 (BMP-2 transgene is an attractive therapeutic modality for the treatment of large bone defects: it provides both stem cells with the ability to form bone and an effective bone inducer while avoiding viral gene transfer. The objective of the present study was to determine the influence of the promoter driving the human BMP-2 gene on the level and duration of BMP-2 expression after transgene electrotransfer into rat MSCs. Cytomegalovirus, elongation factor-1α, glyceraldehyde 3-phosphate dehydrogenase, and beta-actin promoters resulted in a BMP-2 secretion rate increase of 11-, 78-, 66- and 36-fold over respective controls, respectively. In contrast, the osteocalcin promoter had predictable weak activity in undifferentiated MSCs but induced the strongest BMP-2 secretion rates in osteoblastically-differentiated MSCs. Regardless of the promoter driving the transgene, a plateau of maximal BMP-2 secretion persisted for at least 21 d after the hBMP-2 gene electrotransfer. The present study demonstrates the feasibility of gene electrotransfer for efficient BMP-2 transgene delivery into MSCs and for a three-week sustained BMP-2 expression. It also provides the first in vitro evidence for a safe alternative to viral methods that permit efficient BMP-2 gene delivery and expression in MSCs but raise safety concerns that are critical when considering clinical applications.

  15. Construction and characterization of osteogenic and vascular endothelial cell sheets from rat adipose-derived mesenchymal stem cells.

    Science.gov (United States)

    Zhang, Hualin; Yu, Na; Zhou, Yueli; Ma, Hairong; Wang, Juan; Ma, Xuerong; Liu, Jinsong; Huang, Jin; An, Yilin

    2016-10-01

    In this study, adipose-derived mesenchymal stem cells (ADSCs) were isolated from adipose tissues of rats. Flow cytometry identification showed that ADSCs of passage 3 highly expressed CD29 and CD44, but hardly expressed CD31 and CD45. Adipogenic, osteogenic, and chondrogenic differentiation were confirmed by the results of oil red O staining, alkaline phosphatase (ALP), and alcian blue staining, respectively. ADSCs at a density of 1×10(6)/cm(2) were cultured in the osteogenic medium and the osteogenic cell sheets could be obtained after 14 d. The cell sheets were positive with von kossa staining. The transmission electron microscopy (TEM) result showed that needle-like calcium salt crystals were deposited on the ECM. These results suggested that the osteogenic cell sheets may have potential osteogenesis ability. ADSCs at a density of 1×10(6)/cm(2) were cultured in the endothelial cell growth medium-2 and the endothelial cell sheets can be formed after 16 d of culture. The TEM image confirmed that the Weibel-Palade corpuscle was seen in the cells. The expression of CD31 was positive, suggesting that the endothelial cell sheets may have a strong ability to form blood vessels. In this study, two types of cell sheets with the potential abilities of osteogenesis and blood vessels formation were obtained by induced culture of ADSCs in vitro, which lays a foundation to build vascularized tissue engineered bone for the therapy of bone defects.

  16. Cartilage repair by human umbilical cord blood-derived mesenchymal stem cells with different hydrogels in a rat model.

    Science.gov (United States)

    Park, Yong-Beom; Song, Minjung; Lee, Choong-Hee; Kim, Jin-A; Ha, Chul-Won

    2015-11-01

    This study was carried out to assess the feasibility of human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) in articular cartilage repair and to further determine a suitable delivering hydrogel in a rat model. Critical sized full thickness cartilage defects were created. The hUCB-MSCs and three different hydrogel composites (hydrogel A; 4% hyaluronic acid/30% pluronic (1:1, v/v), hydrogel B; 4% hyaluronic acid, and hydrogel C; 4% hyaluronic acid/30% pluronic/chitosan (1:1:2, v/v)) were implanted into the experimental knee (right knee) and hydrogels without hUCB-MSCs were implanted into the control knee (left knee). Defects were evaluated after 8 weeks. The hUCB-MSCs with hydrogels composites resulted in a better repair as seen by gross and histological evaluation compared with hydrogels without hUCB-MSCs. Among the three different hydrogels, the 4% hyaluronic acid hydrogel composite (hydrogel B) showed the best result in cartilage repair as seen by the histological evaluation compared with the other hydrogel composites (hydrogel A and C). The results of this study suggest that hUCB-MSCs may be a promising cell source in combination with 4% hyaluronic acid hydrogels in the in vivo repair of cartilage defects.

  17. Early life ethanol exposure causes long-lasting disturbances in rat mesenchymal stem cells via epigenetic modifications

    Energy Technology Data Exchange (ETDEWEB)

    Leu, Yu-Wei [Department of Life Science and Institute of Molecular Biology, National Chung Cheng University, Chia-Yi 621, Taiwan (China); Chu, Pei-Yi [Department of Pathology, Show Chwan Memorial Hospital, Changhua 500, Taiwan (China); Chen, Chien-Min [Division of Neurosurgery, Changhua Christian Hospital, Changhua 500, Taiwan (China); Yeh, Kun-Tu [Department of Pathology, Changhua Christian Hospital, Changhua 500, Taiwan (China); Liu, Yu Ming; Lee, Yen-Hui; Kuo, Shan-Tsu [Department of Life Science and Institute of Molecular Biology, National Chung Cheng University, Chia-Yi 621, Taiwan (China); Hsiao, Shu-Huei, E-mail: bioshh@ccu.edu.tw [Department of Life Science and Institute of Molecular Biology, National Chung Cheng University, Chia-Yi 621, Taiwan (China)

    2014-10-24

    Highlights: • Ethanol exposure alters proliferation and differentiation of MSCs. • Ethanol exposure suppresses osteogenesis and adipogenesis of MSCs. • H3K27me3-associated genes/pathways are affected in ethanol-exposed MSCs. • Expression of lineage-specific genes is dysregulated in ethanol-exposed MSCs. - Abstract: Fetal alcohol syndrome (FAS) is a birth defect due to maternal alcohol consumption during pregnancy. Because mesenchymal stem cells (MSCs) are the main somatic stem cells in adults and may contribute to tissue homeostasis and repair in adulthood, we investigated whether early life ethanol exposure affects MSCs and contributes to the propensity for disease onset in later life. Using a rodent model of FAS, we found that ethanol exposure (5.25 g/kg/day) from postnatal days 4 to 9 in rat pups (mimic of human third trimester) caused long-term anomalies in bone marrow-derived MSCs. MSCs isolated from ethanol-exposed animals were prone to neural induction but resistant to osteogenic and adipogenic inductions compared to their age-matched controls. The altered differentiation may contribute to the severe trabecular bone loss seen in ethanol-exposed animals at 3 months of age as well as overt growth retardation. Expression of alkaline phosphatase, osteocalcin, aP2, and PPARγ were substantially inhibited, but BDNF was up-regulated in MSCs isolated from ethanol-exposed 3 month-old animals. Several signaling pathways were distorted in ethanol-exposed MSCs via altered trimethylation at histone 3 lysine 27. These results demonstrate that early life ethanol exposure can have long-term impacts in rat MSCs by both genetic and epigenetic mechanisms.

  18. Mechano-growth factor induces migration of rat mesenchymal stem cells by altering its mechanical properties and activating ERK pathway

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    Wu, Jiamin; Wu, Kewen; Lin, Feng; Luo, Qing; Yang, Li; Shi, Yisong [Key Laboratory of Biorheological Science and Technology, Ministry of Education, Bioengineering College, Chongqing University, Chongqing 400044 (China); Song, Guanbin, E-mail: song@cqu.edu.cn [Key Laboratory of Biorheological Science and Technology, Ministry of Education, Bioengineering College, Chongqing University, Chongqing 400044 (China); Sung, Kuo-Li Paul [Key Laboratory of Biorheological Science and Technology, Ministry of Education, Bioengineering College, Chongqing University, Chongqing 400044 (China); Department of Bioengineering, University of California, San Diego, La Jolla, CA 92093-0412 (United States)

    2013-11-08

    Highlights: •MGF induced the migration of rat MSC in a concentration-dependent manner. •MGF enhanced the mechanical properties of rMSC in inducing its migration. •MGF activated the ERK 1/2 signaling pathway of rMSC in inducing its migration. •rMSC mechanics may synergy with ERK 1/2 pathway in MGF-induced rMSC migration. -- Abstract: Mechano-growth factor (MGF) generated by cells in response to mechanical stimulation has been identified as a mechano effector molecule, playing a key role in regulating mesenchymal stem cell (MSC) function, including proliferation and migration. However, the mechanism(s) underlying how MGF-induced MSC migration occurs is still unclear. In the present study, MGF motivated migration of rat MSCs (rMSCs) in a concentration-dependent manner and optimal concentration of MGF at 50 ng/mL (defined as MGF treatment in this paper) was demonstrated. Notably, enhancement of mechanical properties that is pertinent to cell migration, such as cell traction force and cell stiffness were found to respond to MGF treatment. Furthermore, MGF increased phosphorylation of extracellular signal-regulated kinase (ERK), ERK inhibitor (i.e., PD98059) suppressed ERK phosphorylation, and abolished MGF-induced rMSC migration were found, demonstrating that ERK is involved molecule for MGF-induced rMSC migration. These in vitro evidences of MGF-induced rMSC migration and its direct link to altering rMSC mechanics and activating the ERK pathway, uncover the underlying biomechanical and biological mechanisms of MGF-induced rMSC migration, which may help find MGF-based application of MSC in clinical therapeutics.

  19. Pro-bone and antifat effects of green tea and its polyphenol, epigallocatechin, in rat mesenchymal stem cells in vitro.

    Science.gov (United States)

    Ko, Chun Hay; Siu, Wing Sum; Wong, Hing Lok; Shum, Wai Ting; Fung, Kwok Pui; San Lau, Clara Bik; Leung, Ping Chung

    2011-09-28

    Green tea has been demonstrated recently as a potent bone supportive agent. Our previous studies showed that green tea and its polyphenolic constituents can promote bone-forming osteoblast activities and inhibit the bone-resorpting osteoclast formation. The objective of the present study was to investigate whether green tea and its components can regulate the osteogenic and adipogenic differentiation in pluripotent rat mesenchymal stem cells (MSCs). The rat MSCs were isolated from the bone marrow of tibiae and femora. The cells were treated with decaffeinated green tea extract (GTE) and six tea polyphenols under osteogenic induction. The alkaline phosphatase (ALP) activities and matrix calcium (Ca) deposition were assessed after 7 and 14 days of treatment. Our results demonstrated that GTE could significantly increase ALP dose dependently in the concentrations without cytotoxicity (0-100 μg/mL). Among six tested tea polyphenols, epigallocatechin (EGC) was shown to be the most effective in promoting osteogenic differentiation. At 20 μM, EGC increased ALP levels and Ca deposition significantly by 2.3- and 1.7-fold, respectively, when compared with the control group. EGC also increased the mRNA expression of bone formation markers runt-related transcription factor 2, ALP, osteonectin, and osteopontin. Furthermore, EGC demonstrated its antiadipogenicity by decreasing the adipocyte formation and inhibiting the mRNA expression levels of the adipogenic markers peroxisome proliferator-activated receptor γ, ccaat/enhancer-binding protein β, and fatty acid binding protein 4. In conclusion, this is the first report of the dual action of green tea polyphenol EGC in promoting osteogenesis and inhibiting adipocyte formation in MSCs. Our results provide scientific evidence to support the potential use of green tea in supporting the bone against degenerative diseases such as osteoporosis.

  20. Intramyocardial implantation of differentiated rat bone marrow mesenchymal stem cells enhanced by TGF-β1 improves cardiac function in heart failure rats

    Energy Technology Data Exchange (ETDEWEB)

    Lv, Y. [Department of Histology and Embryology, Hebei Medical University, Shijiazhuang, Hebei (China); Liu, B. [Department of Pathology, the First Affiliated Hospital of Hebei North University, Zhangjiakou, Hebei (China); Wang, H.P. [Department of Histology and Embryology, Hebei North University, Zhangjiakou, Hebei (China); Zhang, L. [Department of Histology and Embryology, Hebei Medical University, Shijiazhuang, Hebei (China)

    2016-05-31

    The present study tested the hypotheses that i) transforming growth factor beta 1 (TGF-β1) enhances differentiation of rat bone marrow mesenchymal stem cells (MSCs) towards the cardiomyogenic phenotype and ii) intramyocardial implantation of the TGF-β1-treated MSCs improves cardiac function in heart failure rats. MSCs were treated with different concentrations of TGF-β1 for 72 h, and then morphological characteristics, surface antigens and mRNA expression of several transcription factors were assessed. Intramyocardial implantation of these TGF-β1-treated MSCs to infarcted heart was also investigated. MSCs were initially spindle-shaped with irregular processes. On day 28 after TGF-β1 treatment, MSCs showed fusiform shape, orientating parallel with one another, and were connected with adjoining cells forming myotube-like structures. Immunofluorescence revealed the expression of cardiomyocyte-specific proteins, α-sarcomeric actin and troponin T, in these cells. The mRNA expression of GATA4 and Nkx2.5 genes was slightly increased on day 7, enhanced on day 14 and decreased on day 28 while α-MHC gene was not expressed on day 7, but expressed slightly on day 14 and enhanced on day 28. Transmission electron microscopy showed that the induced cells had myofilaments, z line-like substances, desmosomes, and gap junctions, in contrast with control cells. Furthermore, intramyocardial implantation of TGF-β1-treated MSCs to infarcted heart reduced scar area and increased the number of muscle cells. This structure regeneration was concomitant with the improvement of cardiac function, evidenced by decreased left ventricular end-diastolic pressure, increased left ventricular systolic pressure and increased maximal positive pressure development rate. Taken together, these results indicate that intramyocardial implantation of differentiated MSCs enhanced by TGF-β1 improved cardiac function in heart failure rats.

  1. Exosomes from Human Synovial-Derived Mesenchymal Stem Cells Prevent Glucocorticoid-Induced Osteonecrosis of the Femoral Head in the Rat

    Science.gov (United States)

    Guo, Shang-Chun; Tao, Shi-Cong; Yin, Wen-Jing; Qi, Xin; Sheng, Jia-Gen; Zhang, Chang-Qing

    2016-01-01

    Osteonecrosis of the femoral head (ONFH) represents a debilitating complication following glucocorticoid (GC)-based therapy. Synovial-derived mesenchymal stem cells (SMSCs) can exert protective effect in the animal model of GC-induced ONFH by inducing cell proliferation and preventing cell apoptosis. Recent studies indicate the transplanted cells exert therapeutic effects primarily via a paracrine mechanism and exosomes are an important paracrine factor that can be directly used as therapeutic agents for tissue engineering. Herein, we provided the first demonstration that the early treatment of exosomes secreted by human synovial-derived mesenchymal stem cells (SMSC-Exos) could prevent GC-induced ONFH in the rat model. Using a series of in vitro functional assays, we found that SMSC-Exos could be internalized into bone marrow derived stromal cells (BMSCs) and enhance their proliferation and have anti-apoptotic abilities. Finally, SMSC-Exos may be promising for preventing GC-induced ONFH.

  2. Potential of bone marrow mesenchymal stem cells in management of Alzheimer's disease in female rats.

    Science.gov (United States)

    Salem, Ahmed M; Ahmed, Hanaa H; Atta, Hazem M; Ghazy, Mohamed A; Aglan, Hadeer A

    2014-12-01

    Alzheimer's disease (AD) has been called the disease of the century with significant clinical and socioeconomic impacts. Pharmacological treatment has limited efficacy and only provides symptomatic relief without long-term cure. Accordingly, there is an urgent need to develop novel and effective medications for AD. Stem cell-based therapy is a promising approach to handling neurodegenerative diseases. Therefore, the current study aimed to explore the possible therapeutic role of single intravenous injection of bone marrow derived mesenchymal stem cells (BM-MSCs) after 4 months in management of AD in the experimental model. The work also extended to compare the therapeutic potential of BM-MSCs with 2 conventional therapies of AD; rivastigmine and cerebrolysin administered daily. BM-MSCs were able to home at the injured brains and produced significant increases in the number of positive cells for choline acetyltransferase (ChAT) and survivin expression, as well as selective AD indicator-1 (seladin-1) and nestin gene expression. Histopathological examination indicated that BM-MSCs could remove beta-amyloid plaques from hippocampus. Significant improvement in these biomarkers was similar to or better sometimes than the reference drugs, clearly showing the potential therapeutic role of BM-MSCs against AD through their anti-apoptotic, neurogenic and immunomodulatory properties.

  3. Comparative Study on the Differentiation of Mesenchymal Stem Cells Between Fetal and Postnatal Rat Spinal Cord Niche.

    Science.gov (United States)

    Cao, Songying; Wei, Xiaowei; Li, Hui; Miao, Jianing; Zhao, Guifeng; Wu, Di; Liu, Bo; Zhang, Yi; Gu, Hui; Wang, Lili; Fan, Yang; An, Dong; Yuan, Zhengwei

    2016-01-01

    In a previous study, we established a prenatal surgical approach and transplanted mesenchymal stem cells (MSCs) into the fetal rat spinal column to treat neural tube defects (NTDs). We found that the transplanted MSCs survived and differentiated into neural lineage cells. Various cytokines and extracellular signaling systems in the spinal cord niche play an important role in cell differentiation. In this study, we observed the differentiation of transplanted MSCs in different spinal cord niches and further observed the expression of neurotrophic factors and growth factors in the spinal cord at different developmental stages to explore the mechanism of MSC differentiation in different spinal cord niches. The results showed that transplanted MSCs expressed markers of neural precursor cells (nestin), neurogliocytes (GFAP), and neurons (β-tubulin). The percentages of GFP(+)/nestin(+) double-positive cells in transplanted MSCs in E16, P1, and P21 rats were 18.31%, 12.18%, and 5.06%, respectively. The percentages of GFP(+)/GFAP(+) double-positive cells in E16, P1, and P21 rats were 32.01%, 15.35%, and 12.56%, respectively. The percentages of GFP(+)/β-tubulin(+) double-positive cells in E16, P1, and P21 were 11.76%, 7.62%, and 4.88%, respectively. The differentiation rates of MSCs in embryonic spinal cords were significantly higher than in postnatal spinal cords (p < 0.05). We found that the transplanted MSCs expressed synapsin-1 at different developmental stages. After MSC transplantation, we observed that neurotrophic factor-3 (NT-3), fibroblast growth factor-2 (FGF-2), FGF-8, transforming growth factor-α (TGF-α), vascular endothelial growth factor (VEGF), and platelet-derived growth factor (PDGF) significantly increased in the MSC transplantation group compared with the blank injection group. Furthermore, FGF-2 and VEGF expression were positively correlated with the number of surviving MSCs. In addition, we found that the expression of brain

  4. The Effect of Aligned and Random Electrospun Fibrous Scaffolds on Rat Mesenchymal Stem Cell Proliferation

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    Zahra Zonoubi

    2012-01-01

    Full Text Available Objective: The development of combining mesenchymal stem cells (MSCs with surface modified three-dimensional (3D biomaterial scaffold provides a desirable alternative for replacement of damaged and diseased tissue. Nanofibrous scaffolds serve as suitable environment for cell attachment and proliferation due to their similarity to the physical dimension of the natural extracellular matrix. In this study the properties of plasma treated poly-C-caprolactone nanofiber scaffolds (p-PCL and unaltered PCL scaffolds were compared,and then p-PCL scaffolds were evaluated for MSC culture.Materials and Methods: Aligned and random PCL nanofibrus scaffolds were fabricatedby electrospining and their surface modified with O2 plasma treatment to enhanceMSC proliferation, adhesion and interaction. Chemical and mechanical characterizationswere carried out using scanning electron microscopy (SEM, water contact angle and tensile testing. Cell adhesion and morphology were evaluated using SEM 1 day after culture. Statistical analysis was carried out using one way analysis of variance(ANOVA.Results: The proliferation of MSCs were evaluated using 3-(4,5-Dimethylthiazol-2-yl-2,5-DiphenyltetrazoliumBromide(MTT assay on day 1, 3, and 5 after cell culture. Results showed that the numbers of cells that had grown on PCL nanofibrous scaffolds were significantly higher than those of control surfaces without nanofibers. Furthermore, the proliferation of MSCs on random nanofiber was significantly higher compared to that on aligned nanofiber.Conclusion: This study showed that while both aligned and random plasma treated PCL nanofibrous scaffold are more suitable substrates for MSC growth than tissue culture plates, random nanofiber best supported the proliferation of MSCs.

  5. Effects of magnetic nanoparticle-incorporated human bone marrow-derived mesenchymal stem cells exposed to pulsed electromagnetic fields on injured rat spinal cord.

    Science.gov (United States)

    Cho, Hyunjin; Choi, Yun-Kyong; Lee, Dong Heon; Park, Hee Jung; Seo, Young-Kwon; Jung, Hyun; Kim, Soo-Chan; Kim, Sung-Min; Park, Jung-Keug

    2013-01-01

    Transplanting mesenchymal stem cells into injured lesions is currently under study as a therapeutic approach for spinal cord injury. In this study, the effects of a pulsed electromagnetic field (PEMF) on injured rat spinal cord were investigated in magnetic nanoparticle (MNP)-incorporated human bone marrow-derived mesenchymal stem cells (hBM-MSCs). A histological analysis revealed significant differences in MNP-incorporated cell distribution near the injured site under the PEMF in comparison with that in the control group. We confirmed that MNP-incorporated cells were widely distributed in the lesions under PEMF. The results suggest that MNP-incorporated hBM-MSCs were guided by the PEMF near the injured site, and that PEMF exposure for 8 H per day over 4 weeks promoted behavioral recovery in spinal cord injured rats. The results show that rats with MNP-incorporated hBM-MSCs under a PEMF were more effective on the Basso, Beattie, and Bresnahan behavioral test and suggest that the PEMF enhanced the action of transplanted cells for recovery of the injured lesion.

  6. Effect of advanced oxidation protein products on the proliferation and osteogenic differentiation of rat mesenchymal stem cells.

    Science.gov (United States)

    Sun, Nan; Yang, Li; Li, Yingbin; Zhang, Hua; Chen, Hong; Liu, Duan; Li, Qingnan; Cai, Dehong

    2013-08-01

    Advanced oxidation protein products (AOPPs) as a novel marker of oxidative stress, are involved in a variety of diseases, including osteoporosis. Although a number of studies have shown the possible functions of AOPPs in biological processes, little is known about the role of AOPPs in the pathogenesis of osteoporosis. In this study, we aimed to investigate the effect of AOPPs on the proliferation and osteogenic differentiation of rat mesenchymal stem cells (MSCs). MSCs, isolated from bone marrow, were cultured in the absence or presence of AOPPs (50, 100, 200 and 400 mg/ml). MTT assay was used to determine the proliferative ability of the cells. Alkaline phosphatase (ALP) activity, the mRNA expression of ALP and collagen I and bone nodule formation were detected to assess osteogenic differentiation. Reactive oxygen species (ROS) generation was analyzed with the probe 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA). The expression of receptor of advanced glycation end-products (RAGE) at the mRNA and protein level was detected by real-time PCR and western blot analysis, respectively. Compared with the control group, AOPPs inhibited MSC proliferation in a dose- and time-dependent manner. Moreover, AOPPs induced a significant reduction in ALP activity, as well as a decrease in ALP and collagen I mRNA levels in the MSCs; bone nodule formation was also inhibited. Furthermore, AOPPs increased ROS generation in the MSCs, and upregulated the expression of RAGE at the mRNA and protein level. These results suggest that AOPPs inhibit the proliferation and osteogenic differentiation of MSCs, possibly through the AOPPs-RAGE-ROS pathway; this may be an important mechanism in the development of osteoporosis.

  7. Promoting effect of small molecules in cardiomyogenic and neurogenic differentiation of rat bone marrow-derived mesenchymal stem cells.

    Science.gov (United States)

    Khanabdali, Ramin; Saadat, Anbarieh; Fazilah, Maizatul; Bazli, Khairul Fidaa' Khairul; Qazi, Rida-e-Maria; Khalid, Ramla Sana; Hasan Adli, Durriyyah Sharifah; Moghadamtousi, Soheil Zorofchian; Naeem, Nadia; Khan, Irfan; Salim, Asmat; Shamsuddin, ShamsulAzlin Ahmad; Mohan, Gokula

    2016-01-01

    Small molecules, growth factors, and cytokines have been used to induce differentiation of stem cells into different lineages. Similarly, demethylating agents can trigger differentiation in adult stem cells. Here, we investigated the in vitro differentiation of rat bone marrow mesenchymal stem cells (MSCs) into cardiomyocytes by a demethylating agent, zebularine, as well as neuronal-like cells by β-mercaptoethanol in a growth factor or cytokines-free media. Isolated bone marrow-derived MSCs cultured in Dulbecco's Modified Eagle's Medium exhibited a fibroblast-like morphology. These cells expressed positive markers for CD29, CD44, and CD117 and were negative for CD34 and CD45. After treatment with 1 μM zebularine for 24 hours, the MSCs formed myotube-like structures after 10 days in culture. Expression of cardiac-specific genes showed that treated MSCs expressed significantly higher levels of cardiac troponin-T, Nkx2.5, and GATA-4 compared with untreated cells. Immunocytochemical analysis showed that differentiated cells also expressed cardiac proteins, GATA-4, Nkx 2.5, and cardiac troponin-T. For neuronal differentiation, MSCs were treated with 1 and 10 mM β-mercaptoethanol overnight for 3 hours in complete and serum-free Dulbecco's Modified Eagle's Medium, respectively. Following overnight treatment, neuron-like cells with axonal and dendritic-like projections originating from the cell body toward the neighboring cells were observed in the culture. The mRNA expression of neuronal-specific markers, Map2, Nefl, Tau, and Nestin, was significantly higher, indicating that the treated cells differentiated into neuronal-like cells. Immunostaining showed that differentiated cells were positive for the neuronal markers Flk, Nef, Nestin, and β-tubulin.

  8. Effects of BMP2 and VEGF165 on the osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells.

    Science.gov (United States)

    Lin, Zhaowei; Wang, Jiang-Sheng; Lin, Lijun; Zhang, Jingwen; Liu, Yunlong; Shuai, Ming; Li, Qi

    2014-03-01

    Bone marrow-derived mesenchymal stem cells (MSCs) are dominant seed cell sources for bone regeneration. Bone morphogenetic proteins (BMPs) initiate cartilage and bone formation in a sequential cascade. Vascular endothelial growth factor (VEGF) is an essential coordinator of extracellular matrix remodeling, angiogenesis and bone formation. In the present study, the effects of the vascular endothelial growth factor 165 (VEGF165) and bone morphogenetic protein 2 (BMP2) genes on bone regeneration were investigated by the lentivirus-mediated cotransfection of the two genes into rat bone marrow-derived MSCs. The successful co-expression of the two genes in the MSCs was confirmed using quantitative polymerase chain reaction (qPCR) and western blot analysis. The results of alizarin red and alkaline phosphatase (ALP) staining at 14 days subsequent to transfection showed that the area of staining in cells transfected with BMP2 alone was higher than that in cells transfected with BMP2 and VEGF165 or untransfected control cells, while the BMP2 + VEGF165 group showed significantly more staining than the untransfected control. This indicated that BMP2 alone exhibited a stronger effect in bone regeneration than BMP2 in combination with VEGF165. Similarly, in inducing culture medium, the ALP activity of the BMP2 + VEGF165 group was notably suppressed compared with that of the BMP2 group. The overexpression of VEGF165 inhibited BMP2-induced MSC differentiation and osteogenesis in vitro. Whether or not local VEGF gene therapy is likely to affect bone regeneration in vivo requires further investigation.

  9. 大鼠骨髓间充质干细胞在子宫内膜组织中迁移分化的研究%Study on migration and differentiation of rat mesenchymal stem cells in endometrial tissue

    Institute of Scientific and Technical Information of China (English)

    彭艳; 何援利; 付霞霏; 刘芳

    2012-01-01

    Objective; To observe the migration and differentiation of rat mesenchymal stem cells in endometrial tissue. Methods; The mesenchymal stem cells of the second generation male rats were labeled by PKH26. Five female rats in experimental group were treated with injection of 1 × 107 ml PKH 26 into caudal vein to label mesenchymal stem cells, while five female rats in control group were treated with injection of 1 ml normal saline into caudal vein. Uterine frozen sections were prepared after six weeks, and then the distribution of mesenchymal stem cells in endometrial tissue was observed under fluorescence microscope, the expression of keratin in mesenchymal stem cells was detected by immunofluorescent staining. Results; The mesenchymal stem cells labeled by PKH 26 distributed in endometrial tissue of rats diffusely in experimental group, the expression of keratin was positive in a few mesenchymal stem cells labeled by PKH 26 in glandular epithelium. The mesenchymal stem cells labeled by PKH 26 didn t express in endometrium of rats in control group. Conclusion; The mesenchymal stem cells can migrant to endometrium, differentiate into glandular epithelial cells, and participate in proliferation of endometrial cells.%目的:观察大鼠骨髓间充质于细胞在子宫内膜组织的迁移分化情况.方法:第2代雄性大鼠骨髓间充质于细胞行PKH26标记,实验组5只雌性大鼠尾静脉注射1×107/mlPKH26标记BMSCs 1 ml,对照组5只雌性大鼠尾静脉注射生理盐水1 ml.6周后切下子宫制成冰冻切片,荧光显微镜下观察标记细胞在子宫内膜组织的分布情况,并进行免疫荧光染色检测标记细胞角蛋白的表达.结果:实验组大鼠子宫内膜组织均可见标记细胞散在分布于子宫内膜组织中,腺上皮中少数标记细胞角蛋白染色阳性.对照组子宫内膜均未见标记细胞.结论:BMSCs可以迁移至子宫内膜,可以向腺上皮细胞分化,参与子宫内膜的增生.

  10. 5-Azacytidine prevents cisplatin induced nephrotoxicity and potentiates anticancer activity of cisplatin by involving inhibition of metallothionein, pAKT and DNMT1 expression in chemical induced cancer rats.

    Science.gov (United States)

    Tikoo, Kulbhushan; Ali, Idrish Yunus; Gupta, Jeena; Gupta, Chanchal

    2009-12-15

    5-Azactydine inhibits cell growth by direct cytotoxic action as well as by inhibition of DNA methyl transferase enzyme. Inhibitors of DNMT have been reported to potentiate the therapeutic activity of cisplatin in vitro. Dose dependent bone marrow toxicity, neurotoxicity and nephrotoxicity are the major side effects of cisplatin, limiting its use as an effective chemotherapeutic agent. The present study was aimed to reduce the nephrotoxic potential of cisplatin without compensating its potency. To best of our knowledge, this is the first report which shows that the combination of 5-azacytidine with cisplatin leads to remarkable reduction in nephrotoxicity, by involving inhibition of cisplatin induced metallothionein expression. 5-Azacytidine treatment with cisplatin leads to maximum reduction in tumor size in DMH induced colon cancer and tumor volume in DMBA induced breast cancer bearing SD rats. This combination regimen prevents phosphorylation and acetylation of histone H3 which may be involved in inhibition of aberrant gene expression in colon tumors. Further, 5-azacytidine potentiated cisplatin induced antitumor activity by involving decreased expression of pAKT, DNMT1 and an increased expression of p38 in colon tumors. Thus, combination of 5-azactydine with cisplatin attenuates the cisplatin induced nephrotoxicity and potentiates the anti-cancer activity which can have profound clinical implications.

  11. Chemically induced mouse models of intestinal inflammation.

    Science.gov (United States)

    Wirtz, Stefan; Neufert, Clemens; Weigmann, Benno; Neurath, Markus F

    2007-01-01

    Animal models of intestinal inflammation are indispensable for our understanding of the pathogenesis of Crohn disease and ulcerative colitis, the two major forms of inflammatory bowel disease in humans. Here, we provide protocols for establishing murine 2,4,6-trinitro benzene sulfonic acid (TNBS)-, oxazolone- and both acute and chronic dextran sodium sulfate (DSS) colitis, the most widely used chemically induced models of intestinal inflammation. In the former two models, colitis is induced by intrarectal administration of the covalently reactive reagents TNBS/oxazolone, which are believed to induce a T-cell-mediated response against hapten-modified autologous proteins/luminal antigens. In the DSS model, mice are subjected several days to drinking water supplemented with DSS, which seems to be directly toxic to colonic epithelial cells of the basal crypts. The procedures for the hapten models of colitis and acute DSS colitis can be accomplished in about 2 weeks but the protocol for chronic DSS colitis takes about 2 months.

  12. Adipose derived mesenchymal stem cells transplantation via portal vein improves microcirculation and ameliorates liver fibrosis induced by CCl4 in rats

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    Wang Yu

    2012-06-01

    Full Text Available Abstract Introduction Adipose derived mesenchymal stem cells (ADMSCs, carrying the similar characteristics to bone marrow mesenchymal stem cells, only much more abundant and easier to obtain, may be a promising treatment for liver fibrosis. We aim to investigate the therapeutic potential of ADMSCs transplantation in liver fibrosis caused by carbon tetrachloride (CCl4 in rats as well as its underlying mechanism, and to further explore the appropriate infusion pathway. Methods ADMSCs were isolated, cultured and identified. Placebo and ADMSCs were transplanted via portal vein and tail vein respectively into carbon tetrachloride (CCl4-induced liver fibrosis rats. Computed tomography (CT perfusion scan and microvessel counts were performed to measure the alteration of liver microcirculation after therapy. Liver function tests and histological findings were estimated. Results CT perfusion scan shown significant decrease of hepatic arterial perfusion index, significant increased portal vein perfusion, total liver perfusion in rats receiving ADMSCs from portal vein, and Factor VIII (FVIII immunohistochemical staining shown significant decrease of microvessels in rats receiving ADMSCs from portal vein, indicating microcirculation improvement in portal vein group. Vascular endothelial growth Factor (VEGF was significantly up-regulated in fibrosis models, and decreased after ADMSCs intraportal transplantation. A significant improvement of liver functional test and histological findings in portal vein group were observed. No significance was found in rats receiving ADMSCs from tail vein. Conclusions ADMSCs have a therapeutic effect against CCl4-mediated liver fibrosis. ADMSCs may benefit the fibrotic liver through alteration of microcirculation, evidenced by CT perfusion scan and down-regulation of VEGF. Intraportal transplantation is a better pathway than tail vein transplantation.

  13. Age-related changes in rat bone-marrow mesenchymal stem cell plasticity

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    Chase P Bryant

    2011-10-01

    Full Text Available Abstract Background The efficacy of adult stem cells is known to be compromised as a function of age. This therefore raises questions about the effectiveness of autologous cell therapy in elderly patients. Results We demonstrated that the expression profile of stemness markers was altered in BM-MSCs derived from old rats. BM-MSCs from young rats (4 months expressed Oct-4, Sox-2 and NANOG, but we failed to detect Sox-2 and NANOG in BM-MSCs from older animals (15 months. Chondrogenic, osteogenic and adipogenic potential is compromised in old BM-MSCs. Stimulation with a cocktail mixture of bone morphogenetic protein (BMP-2, fibroblast growth factor (FGF-2 and insulin-like growth factor (IGF-1 induced cardiomyogenesis in young BM-MSCs but not old BM-MSCs. Significant differences in the expression of gap junction protein connexin-43 were observed between young and old BM-MSCs. Young and old BM-MSCs fused with neonatal ventricular cardiomyocytes in co-culture and expressed key cardiac transcription factors and structural proteins. Cells from old animals expressed significantly lower levels of VEGF, IGF, EGF, and G-CSF. Significantly higher levels of DNA double strand break marker γ-H2AX and diminished levels of telomerase activity were observed in old BM-MSCs. Conclusion The results suggest age related differences in the differentiation capacity of BM-MSCs. These changes may affect the efficacy of BM-MSCs for use in stem cell therapy.

  14. Wnt3a signaling promotes proliferation,myogenic differentiation,and migration of rat bone marrow mesenchymal stem cells

    Institute of Scientific and Technical Information of China (English)

    Yan-chang SHANG; Shu-hui WANG; Fu XIONG; Cui-ping ZHAO; Fu-ning PENG; Shan-wei FENG; Mei-shan LI; Yong LI; Cheng ZHANG

    2007-01-01

    Aim:To investigate the effects of the wingless-related MMTV integration site 3A (Wnt3a) signaling on the proliferation,migration,and the myogenic and adipogenic differentiation of rat bone marrow mesenchymal stem cells (rMSC). Methods:Primary MSC were isolated and cultured from Spragne-Dawley rats and characterized by flow cytometry. Mouse L cells were transfected with Wnt3a cDNA,and conditioned media containing active Wnt3a proteins were prepared. Cell proliferation was evaluated by cell count and 5-bromodeoxyuridine incorporation assay.The migration of rMSC was performed by using a transwell migration and wound healing assay. The myogenic and adipogenic differentiation in rMSC were examined by light microscopy,immunofluorescence,and RT-PCR at different time points after myogenic or adipogenic introduction. Results:Wnt3a signaling induced β-catenin nuclear translocation and activated the Writ pathway in rMSC.In the presence of Wnt3a,rMSC proliferated more rapidly than the control cells,keeping their differentiation potential. Moreover,Wnt3a signaling induced 2.62%and 3.76% of rMSC-expressed desmin and myosin heavy chain after being cultured in myogenic medium. The myogenic differentiation genes,including Pax7,MyoD,Myf5,Myf4,and myogenin,were activated after Wnt3a treatment. On the other hand,Wnt3a inhibited the adipogenic differentiation in rMSC through the downregulated expression of CCAAT/enhancer-binding protein alpha (C/EBPalpha)and peroxisome proliferator-activaled receptor gamma (PPARgamma). Furthermore,Wnt3a promoted the migration capacity of rMSC. Conclusion:The results indicate that Wnt3a signaling can induce myogenic differentiation in rMSC. Wnt3a signaling is also involved in the regulation of the proliferation and migration of rMSC. These results could provide a rational foundation for cell-based tissue repair in humans.

  15. Brain-derived neurotrophic factor genes transfect rat bone marrow mesenchymal stem cells based on cationic polymer vector

    Institute of Scientific and Technical Information of China (English)

    Zunsheng Zhang; Kun Zan; Yonghai Liu; Xia Shen

    2009-01-01

    BACKGROUND: Gene therapy is an effective expression of genes within target cells after transferring exogenous target genes. Both vector selection and transfection method are important factors for gene transfection. An ideal gene vector is required for a high transfusion of target gene and an exact introduction of target gene into specific target cells so as to express gene products. OBJECTIVE: To study the expression of mRNA and protein after transfecting rat bone marrow mesenchymal stem cells (BMSCs) with brain-derived neurotrophic factor (BDNF) genes based on cationic polymer vector. DESIGN, TIME AND SETTING: A randomized, controlled in vitro study using gene engineering, performed at the Neurobiology Laboratory, Xuzhou Medical College between October 2007 and April 2008. MATERIALS: PcDNA3.1 BDNF was obtained from Youbiai Biotechnological Company, Beijing and cationic polymer vector used was the SofastTM gene transfection reagent that was made by Taiyangma Biotechnological Co., Ltd., Xiamen. METHODS: BMSCs extracted from six Sprague Dawley (SD) rats aged 1 month were isolated and cultured in vitro. Third passage BMSCs were inoculated on a 6-well culture plate at the density of 1×106 cells/L. At about 80% confluence, BMSCs were transfected with PcDNA3.1-BDNF (2 μg) combined with SofastTM gene transfection reagent (6 μg) (BDNF group) or with PcDNA3.1 (2 μg) combined with SofastTM gene transfection reagent (6 μg) (blank vector group). Cells that were not transfected with any reagents but still cultured under primary culture conditions were used as a non-transfection group.MAIN OUTCOME MEASURES: Enzyme linked immunosorbent assay was used to measure time efficiency of BMSC-secreted BDNF protein. Twenty-four hours after gene transfection, RT-PCR was used to detect expression of BDNF mRNA in the BMSCs. Immunohistochemistry was used to determine expression of BDNF protein in the BMSCs.RESULTS: BDNF protein expression was detected at day 1 after gene transfection

  16. Dietary extra-virgin olive oil and corn oil differentially modulate the mRNA expression of xenobiotic-metabolizing enzymes in the liver and in the mammary gland in a rat chemically induced breast cancer model.

    Science.gov (United States)

    Manzanares, Miguel Á; Solanas, Montserrat; Moral, Raquel; Escrich, Raquel; Vela, Elena; Costa, Irmgard; Escrich, Eduard

    2015-05-01

    High extra-virgin olive oil (EVOO) and corn oil diets differentially modulate experimental mammary carcinogenesis. We have investigated their influence on the initiation stage through the modulation of the expression of xenobiotic-metabolizing enzymes (XMEs) in the liver and the mammary gland. Female Sprague-Dawley rats were fed a low-fat (LF), high corn oil (HCO), or high EVOO (HOO) diet from weaning and gavaged with 7,12-dimethylbenz(a)anthracene (DMBA). The HCO diet increased the mRNA levels of the phase I enzymes CYP1A1, CYP1A2 and, to a lesser extent, CYP1B1, in the liver. The Aryl hydrocarbon receptor (AhR) seemed to be involved in this upregulated CYP1 expression. However, a slight trend toward an increase in the mRNA levels of the phase II enzymes GSTP1 and NQO1 was observed with the HOO diet. At least in the case of GSTP1, this effect was linked to an increased Nrf2 transactivation activity. This different regulation of the XMEs expression led, in the case of the HCO diet, to a balance between the production of active carcinogenic compounds and their inactivation tilted toward phase I, which would stimulate DMBA-induced cancer initiation, whereas the HOO diet was associated with a slower phase I metabolism accompanied by a faster phase II detoxification, thus reducing the output of the active compounds to the target tissues. In the mammary gland, the differential effects of diets may be conditioned by the state of cell differentiation, sexual maturity, and hormone metabolism.

  17. In Vitro Study of the Effect of Vitamin E on Viability, Morphological Changes and Induction of Osteogenic Differentiation in Adult Rat Bone Marrow Mesenchymal Stem Cells

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    M Soleimani Mehranjani

    2014-10-01

    Full Text Available Introduction: Vitamin E as a strong antioxidant plays an important role in inhibiting free radicals. Therefore, this study aimed to investigate the effect of vitamin E on the viability, morphology and osteogenic differentiation in bone marrow mesenchymal stem cells of an adult rat. Methods: The bone marrow mesenchymal stem cells were extracted using the flashing-out method. At the end of the third passage, cells were divided into groups of control and experimental. Experimental cells were treated withVitamin E (5,10,15,25,50,100,150μM for a period of 21 days in the osteogenic media containing 10% of fetal bovine serum. The cell viability, bone matrix mineralization, intercellular and extracellular calcium deposition, alkaline phosphatase activity, expression of genes and synthesis of proteins of osteopontin and osteocalcin as well as morphological changes of the cells were investigated. The study data was analyzed using one-way ANOVA and T-Test setting the significant P value at P<0.05. Results: Within vitamin- E treated cells, the mean viability, mean bone matrix mineralization, calcium deposition, alkaline phosphatase activity, expression and synthesis of osteopontin and osteocalcin of the mesenchymal stem cells treated with vitamin E significantly increased in a dose dependent manner. Also cytoplasm extensions were observed in the cells treated with vitamin E. Conclusion: Since vitamin E caused a significant increase in cell viability and osteogenic differentiation in the mesenchymal stem cells, therefore it can be utilized in order to increase cell differentiation and cell survival.

  18. Optimal time for mesenchymal stem cell transplantation in rats with myocardial infarction

    Institute of Scientific and Technical Information of China (English)

    Chen-yang JIANG; Chun GUI; Ai-na HE; Xin-yang HU; Jie CHEN; Yun JIANG; Jian-an WANG

    2008-01-01

    Background:Bone marrow mesenehymal stem cell(MSC)transplantation is a promising strategy in the treatment of myocardial infarction(MI).However,the time for transplanting cells remains controversial.The aim of this study was to find an optimal time point for cell transplantation.Methods:MSCs were isolated and cultured from Sprague-Dawley(SD) rats.MI model was set up in SD rats by permanent ligation of left anterior descending coronary artery.MSCs were directly injected into the infarct berder zone at 1 h,1 week and 2 weeks after MI,respectively.Sham-operated and MI centrel groups received equal volume of phosphate buffered saline(PBS).At 4 weeks after MI,cardiac function Was assessed by echocardiography;vessel density Was analyzed on hematoxylin-eosin stained slides by light microscopy;the apoptosis of cardiomyocytes Was evaluated by terminal deoxynucleotidy1 transferase-mediated dUTP nick end-labeling(TUNEL) assay;the expressions of proteins were analyzed by Western blot.Results:MSC transplantation improved cardiac function.reduced the apoptosis of cardiomyocytes and increased vessel density.These benefits were more obvious in l-week group than in 1-h and 2-week groups.There are more obvious increases in the ratio of bc1-2/bax and the expression of vascular endothelial growth factor(VEGF)and more obvious decreases in the expression of cleaved-caspase-3 in 1-week group than those in other two groups.Conclusion:MSC transplantation was beneficial for the recovery of cardiac function.MSC transplantation at l week post-MI exerted the best effects on increases of cardiac function,anti-apoptosis and angiogenesis.

  19. Deferasirox induces mesenchymal-epithelial transition in crocidolite-induced mesothelial carcinogenesis in rats.

    Science.gov (United States)

    Nagai, Hirotaka; Okazaki, Yasumasa; Chew, Shan Hwu; Misawa, Nobuaki; Yasui, Hiroyuki; Toyokuni, Shinya

    2013-11-01

    Asbestos was used worldwide in huge quantities in the past century. However, because of the unexpected carcinogenicity to mesothelial cells with an extremely long incubation period, many countries face this long-lasting social problem. Mesothelioma is often diagnosed in an advanced stage, for which no effective therapeutic protocols are yet established. We previously reported on the basis of animal experiments that the major pathology in asbestos-induced mesothelial carcinogenesis is local iron overload. Here, we undertook to find an effective strategy to prevent, delay, or lower the malignant potential of mesothelioma during asbestos-induced carcinogenesis. We used intraperitoneal injections of crocidolite to rats. We carried out a 16-week study to seek the maximal-tolerated intervention for iron reduction via oral deferasirox administration or intensive phlebotomy. Splenic iron deposition was significantly decreased with either method, and we found that Perls' iron staining in spleen is a good indicator for iron reduction. We injected a total of 10 mg crocidolite at the age of six weeks, and the preventive measures were via repeated oral administration of 25 to 50 mg/kg/d deferasirox or weekly to bimonthly phlebotomy of 4 to 10 mL/kg/d. The animals were observed until 110 weeks. Deferasirox administration significantly increased the fraction of less malignant epithelioid subtype. Although we found a slightly prolonged survival in deferasirox-treated female rats, larger sample size and refinement of the current protocol are necessary to deduce the cancer-preventive effects of deferasirox. Still, our results suggest deferasirox serves as a potential preventive strategy in people already exposed to asbestos via iron reduction.

  20. The effect of progesterone in the prevention of the chemically induced experimental colitis in rats Efeito da progesterona na prevenção de colite experimental induzida quimicamente em ratos

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    Oguzhan Karatepe

    2012-01-01

    Full Text Available PURPOSE: To study the effects of progesterone on an experimental colitis model. METHODS: Wistar albino rats were treated subcutaneously with 2mg/kg once a day during seven days Colitis was induced by intrarectal administration of 5mg trinitrobenzene sulfonic acid (TNBS. Disease activities, macroscopic and microscopic scores were evaluated. To determine the response provoked by progesterone we measured Colonic malondialdehyde (MDA, TNF alfa, IL-6 and Nitric oxide (NO levels in addition to the MPO (Myeloperoxidase and caspase-3 activities. RESULTS: Progesterone ameliorated significantly the macroscopic and microscopic scores. TNBS-induced colitis significantly increased the colonic MDA levels and caspase-3 activities in group 2 in comparison to the control group. The results of the study revealed a decline in MDA, NO, IL6 and TNF-α levels in the colon tissue and in blood due to progesterone therapy in group 3 when compared to the group 2, a significant improvement. Progesterone treatment was associated with decreased MDA, MPO, TNF alfa and caspase-3 activity. CONCLUSION: Progesterone therapy decreased oxidative damage in the colonic mucosa.OBJETIVO: Investigar os efeitos da progesterona em um modelo de colite experimental. MÉTODOS: Ratos albinos Wistar foram tratados subcutaneamente com 2mg/kg por dia durante sete dias. A colite foi induzida por administração intrarretal de 5mg ácido sulfônico trinitrobenzeno (TNBS. Foram avaliadas as atividades da doença, escores macroscópicos e microscópicos Para determinar a resposta provocada pela progesterona foi medida no cólon os níveis de malondialdeído (MDA, TNF alfa, IL-6 e óxido nítrico (NO, além da atividade da MPO (Myeloperoxidase e caspase-3. RESULTADOS: A progesterone melhorou significantemente os escores macroscópicos e microscópicos. A colite induzida pelo TNBS significantemente aumentou os níveis colônicos de MDA e a atividade da caspase-3 no grupo 2 em comparação com o grupo

  1. Neuronal-like differentiation of bone marrow-derived mesenchymal stem cells induced by striatal extracts from a rat model of Parkinson's disease

    Institute of Scientific and Technical Information of China (English)

    Xiaoling Qin; Wang Han; Zhigang Yu

    2012-01-01

    A rat model of Parkinson's disease was established by 6-hydroxydopamine injection into the medial forebrain bundle. Bone marrow-derived mesenchymal stem cells (BMSCs) were isolated from the femur and tibia, and were co-cultured with 10% and 60% lesioned or intact striatal extracts. The results showed that when exposed to lesioned striatal extracts, BMSCs developed bipolar or multi-polar morphologies, and there was an increase in the percentage of cells that expressed glial fibrillary acidic protein (GFAP), nestin and neuron-specific enolase (NSE). Moreover, the percentage of NSE-positive cells increased with increasing concentrations of lesioned striatal extracts. However, intact striatal extracts only increased the percentage of GFAP-positive cells. The findings suggest that striatal extracts from Parkinson's disease rats induce BMSCs to differentiate into neuronal-like cells in vitro.

  2. A role for Gcn5 in cardiomyocyte differentiation of rat mesenchymal stem cells.

    Science.gov (United States)

    Li, Li; Zhu, Jing; Tian, Jie; Liu, Xiaoyan; Feng, Chuan

    2010-12-01

    MSCs possess the capacity of self-renewal and potential of differentiation into various kinds of specialized tissue cells including myocardiocytes. From self-renewing to oriented differentiation, chromatin is remodeled into heritable states that allow activation or maintain the repression of regulatory genes, which means specific genes in self-renewing switched off and specific genes in oriented differentiation activated (Bernstein et al. Cell 125:315-326, 2006). These epigenetic states are established and controlled largely by specific patterns of histone posttranslational modifications, in particular, histone acetylation (Li Nat Rev Genet 3:662-673, 2002). In cardiomyocyte differentiation of rat MSCs, we focused on Gcn5, which linked a known transcriptional coactivator with catalytic histone acetyltransferase activity (Brownell et al. Cell 84:843-851, 1996). To clarify participatory in vivo role of Gcn5, using an RNA interference (RNAi) strategy employing shRNA to specifically knockdown Gcn5 expression in MSCs, we found that HAT activity altered dynamically depended on the inhibition of Gcn5 during MSCs differentiation. Chromatin immunoprecipitation (ChIP) assay showed the increased binding of acetyl histone H3 to the early cardiomyocyte-specific genes GATA4 and NKx2.5 promoters in cardiomyocyte differentiation of MSCs by 5-azacytidine inducing, whereas the decreased binding with lower Gcn5 expression. Cell ultrastructure analysis revealed that MSCs induced by 5-azacytidine possess morphological characteristics of cardiomyocyte cells. The shape of MSCs transfected by Gcn5 RNAi was similar to normal MSCs, but the chromatin showed heavy electron-density and a hard-packed structure. This intermediate state of chromatin may be an inactive part of MSCs differentiation. These results demonstrate that Gcn5, possessing acetyltransferase activity, is involved in regulating chromatin configuration around GATA4 and NKx2.5 in cardiomyocyte differentiation of rat MSCs by

  3. Retinal Electrophysiological Effects of Intravitreal Bone Marrow Derived Mesenchymal Stem Cells in Streptozotocin Induced Diabetic Rats.

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    Eren Çerman

    Full Text Available Diabetic retinopathy is the most common cause of legal blindness in developed countries at middle age adults. In this study diabetes was induced by streptozotocin (STZ in male Wistar albino rats. After 3 months of diabetes, rights eye were injected intravitreally with green fluorescein protein (GFP labelled bone marrow derived stem cells (BMSC and left eyes with balanced salt solution (Sham. Animals were grouped as Baseline (n = 51, Diabetic (n = 45, Diabetic+BMSC (n = 45 eyes, Diabetic+Sham (n = 45 eyes, Healthy+BMSC (n = 6 eyes, Healthy+Sham (n = 6 eyes. Immunohistology analysis showed an increased retinal gliosis in the Diabetic group, compared to Baseline group, which was assessed with GFAP and vimentin expression. In the immunofluorescence analysis BMSC were observed to integrate mostly into the inner retina and expressing GFP. Diabetic group had prominently lower oscillatory potential wave amplitudes than the Baseline group. Three weeks after intravitreal injection Diabetic+BMSC group had significantly better amplitudes than the Diabetic+Sham group. Taken together intravitreal BMSC were thought to improve visual function.

  4. Cooperation by Fibroblasts and Bone Marrow-Mesenchymal Stem Cells to Improve Pancreatic Rat-to-Mouse Islet Xenotransplantation

    Science.gov (United States)

    Meana, Alvaro; Otero, Jesus; Esteban, Manuel M.

    2013-01-01

    Experimental and clinical experiences highlight the need to review some aspects of islet transplantation, especially with regard to site of grafting and control of the immune response. The subcutaneous space could be a good alternative to liver but its sparse vasculature is its main limitation. Induction of graft tolerance by using cells with immunoregulatory properties is a promising approach to avoid graft rejection. Both Fibroblasts and Mesenchymal Stem Cells (MSCs) have shown pro-angiogenic and immunomodulatory properties. Transplantation of islets into the subcutaneous space using plasma as scaffold and supplemented with fibroblasts and/or Bone Marrow-MSCs could be a promising strategy to achieve a functional extra-hepatic islet graft, without using immunosuppressive drugs. Xenogenic rat islets, autologous fibroblasts and/or allogenic BM-MSCs, were mixed with plasma, and coagulation was induced to constitute a Plasma-based Scaffold containing Islets (PSI), which was transplanted subcutaneously both in immunodeficient and immunocompetent diabetic mice. In immunodeficient diabetic mice, PSI itself allowed hyperglycemia reversion temporarily, but the presence of pro-angiogenic cells (fibroblasts or BM-MSCs) within PSI was necessary to improve graft re-vascularization and, thus, consistently maintain normoglycemia. In immunocompetent diabetic mice, only PSI containing BM-MSCs, but not those containing fibroblasts, normalized glycemia lasting up to one week after transplantation. Interestingly, when PSI contained both fibroblasts and BM-MSCs, the normoglycemia period showed an increase of 4-times with a physiological-like response in functional tests. Histology of immunocompetent mice showed an attenuation of the immune response in those grafts with BM-MSCs, which was improved by co-transplantation with fibroblasts, since they increased BM-MSC survival. In summary, fibroblasts and BM-MSCs showed similar pro-angiogenic properties in this model of islet

  5. Effects of umbilical cord tissue mesenchymal stem cells (UCX® on rat sciatic nerve regeneration after neurotmesis injuries

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    Gärtner A

    2013-04-01

    Full Text Available Peripheral nerves have the intrinsic capacity of self-regeneration after traumatic injury but the extent of the regeneration is often very poor. Increasing evidence demonstrates that mesenchymal stem/stromal cells (MSCs may play an important role in tissue regeneration through the secretion of soluble trophic factors that enhance and assist in repair by paracrine activation of surrounding cells. In the present study, the therapeutic value of a population of umbilical cord tissue-derived MSCs, obtained by a proprietary method (UCX®, was evaluated on end-to-end rat sciatic nerve repair. Furthermore, in order to promote both, end-to-end nerve fiber contacts and MSC cell-cell interaction, as well as reduce the flush away effect of the cells after administration, a commercially available haemostatic sealant, Floseal®, was used as vehicle. Both, functional and morphologic recoveries were evaluated along the healing period using extensor postural thrust (EPT, withdrawal reflex latency (WRL, ankle kinematics analysis, and either histological analysis or stereology, in the hyper-acute, acute and chronic phases of healing. The histological analysis of the hyper-acute and acute phase studies revealed that in the group treated with UCX ® alone the Wallerian degeneration was improved for the subsequent process of regeneration, the fiber organization was higher, and the extent of fibrosis was lower. The chronic phase experimental groups revealed that treatment with UCX® induced an increased number of regenerated fibers and thickening of the myelin sheet. Kinematics analysis showed that the ankle joint angle determined for untreated animals was significantly different from any of the treated groups at the instant of initial contact (IC. At opposite toe off (OT and heel rise (HR, differences were found between untreated animals and the groups treated with either UCX® alone or UCX® administered with Floseal®. Overall, the UCX® application presented

  6. Cooperation by fibroblasts and bone marrow-mesenchymal stem cells to improve pancreatic rat-to-mouse islet xenotransplantation.

    Directory of Open Access Journals (Sweden)

    Marcos Perez-Basterrechea

    Full Text Available Experimental and clinical experiences highlight the need to review some aspects of islet transplantation, especially with regard to site of grafting and control of the immune response. The subcutaneous space could be a good alternative to liver but its sparse vasculature is its main limitation. Induction of graft tolerance by using cells with immunoregulatory properties is a promising approach to avoid graft rejection. Both Fibroblasts and Mesenchymal Stem Cells (MSCs have shown pro-angiogenic and immunomodulatory properties. Transplantation of islets into the subcutaneous space using plasma as scaffold and supplemented with fibroblasts and/or Bone Marrow-MSCs could be a promising strategy to achieve a functional extra-hepatic islet graft, without using immunosuppressive drugs. Xenogenic rat islets, autologous fibroblasts and/or allogenic BM-MSCs, were mixed with plasma, and coagulation was induced to constitute a Plasma-based Scaffold containing Islets (PSI, which was transplanted subcutaneously both in immunodeficient and immunocompetent diabetic mice. In immunodeficient diabetic mice, PSI itself allowed hyperglycemia reversion temporarily, but the presence of pro-angiogenic cells (fibroblasts or BM-MSCs within PSI was necessary to improve graft re-vascularization and, thus, consistently maintain normoglycemia. In immunocompetent diabetic mice, only PSI containing BM-MSCs, but not those containing fibroblasts, normalized glycemia lasting up to one week after transplantation. Interestingly, when PSI contained both fibroblasts and BM-MSCs, the normoglycemia period showed an increase of 4-times with a physiological-like response in functional tests. Histology of immunocompetent mice showed an attenuation of the immune response in those grafts with BM-MSCs, which was improved by co-transplantation with fibroblasts, since they increased BM-MSC survival. In summary, fibroblasts and BM-MSCs showed similar pro-angiogenic properties in this model of

  7. Promoting effect of small molecules in cardiomyogenic and neurogenic differentiation of rat bone marrow-derived mesenchymal stem cells

    Directory of Open Access Journals (Sweden)

    Khanabdali R

    2015-12-01

    Full Text Available Ramin Khanabdali,1 Anbarieh Saadat,1 Maizatul Fazilah,1 Khairul Fidaa’ Khairul Bazli,1 Rida-e-Maria Qazi,2 Ramla Sana Khalid,2 Durriyyah Sharifah Hasan Adli,1 Soheil Zorofchian Moghadamtousi,1 Nadia Naeem,2 Irfan Khan,2 Asmat Salim,2 ShamsulAzlin Ahmad Shamsuddin,1 Gokula Mohan1 1Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia; 2Dr Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan Abstract: Small molecules, growth factors, and cytokines have been used to induce differentiation of stem cells into different lineages. Similarly, demethylating agents can trigger differentiation in adult stem cells. Here, we investigated the in vitro differentiation of rat bone marrow mesenchymal stem cells (MSCs into cardiomyocytes by a demethylating agent, zebularine, as well as neuronal-like cells by β-mercaptoethanol in a growth factor or cytokines-free media. Isolated bone marrow-derived MSCs cultured in Dulbecco’s Modified Eagle’s Medium exhibited a fibroblast-like morphology. These cells expressed positive markers for CD29, CD44, and CD117 and were negative for CD34 and CD45. After treatment with 1 µM zebularine for 24 hours, the MSCs formed myotube-like structures after 10 days in culture. Expression of cardiac-specific genes showed that treated MSCs expressed significantly higher levels of cardiac troponin-T, Nkx2.5, and GATA-4 compared with untreated cells. Immunocytochemical analysis showed that differentiated cells also expressed cardiac proteins, GATA-4, Nkx 2.5, and cardiac troponin-T. For neuronal differentiation, MSCs were treated with 1 and 10 mM β-mercaptoethanol overnight for 3 hours in complete and serum-free Dulbecco’s Modified Eagle’s Medium, respectively. Following overnight treatment, neuron-like cells with axonal and dendritic-like projections originating from the

  8. Chemically Induced Damage to the Hippocampal Formation,

    Science.gov (United States)

    1986-05-01

    group for this thesis. % My thanks are also extended to Dr Charles N Allen for collaboration and discussions, to Mrs Anna Heggemsnes and Mrs Jorunn...Trimethyltin Liv UNNi NAALSUND, CHARLES N. ALLEN’ AND FRODE FONNUM Norwegian Defense Research Establishment, Division for Environmental Toxicology, P.O...in the Swartzwelder HS, Hepler J, Holahan W, KingS E, rat studied by silver impregnation. J Camp Leverenz HA. Miller PA and Myers RD. Im- Neural 1972

  9. Therapeutic effects of human amnion-derived mesenchymal stem cell transplantation and conditioned medium enema in rats with trinitrobenzene sulfonic acid-induced colitis

    Science.gov (United States)

    Miyamoto, Shuichi; Ohnishi, Shunsuke; Onishi, Reizo; Tsuchiya, Ikuki; Hosono, Hidetaka; Katsurada, Takehiko; Yamahara, Kenichi; Takeda, Hiroshi; Sakamoto, Naoya

    2017-01-01

    Cell therapy with mesenchymal stem cells (MSCs) is expected to provide a new strategy for the treatment of inflammatory bowel disease (IBD). Large amounts of MSCs can be obtained from human amnion. Therefore, we investigated the effect of transplantation of human amnion-derived MSCs (hAMSCs) or enema of conditioned medium (CM) from hAMSCs into rats with 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis. In the first experiment, 10-week-old male Sprague-Dawley rats were intravenously injected with hAMSCs (1 × 106 cells) 3 h after rectal administration of TNBS (45 mg/kg). In the second experiment, rats with TNBS-induced colitis received CM by enema into the colon for 3 days. Colitis was investigated by endoscopy, histology, immunohistochemistry, and by measuring mRNA expression of inflammatory mediators. Administration of hAMSCs or CM enema significantly improved the endoscopic score. In addition, these two interventions resulted in significantly decreased infiltration of neutrophils and monocytes/macrophages and decreased expression levels of TNF-α, CXCL1, and CCL2. In conclusion, transplantation of hAMSCs and CM enema provided significant improvement in rats with TNBS-induced colitis. CM from hAMSCs and hAMSCs may be new strategies for the treatment of IBD.

  10. [Effect of laminar shear stress on the expression of matrix metalloproteinases-9 in rat bone marrow-derived mesenchymal stem cells].

    Science.gov (United States)

    Chen, Longju; Sun, Xiaodong; Tang, Jie; Ding, Yan; Li, Jing; Li, Wenchun; Gong, Jian; Wang, Hanqin

    2010-12-01

    This paper was designed to investigate the effect of laminar shear stress on matrix metalloproteinase -9 (MMP-9) expression in rat bone marrow-derived mesenchymal stem cells (MSCs), and the possible signal transduction mechanism involved. Rat bone marrow MSCs were isolated and cultured, then, exposed to laminar shear stress at indicated strengths such as low (5dyne/cm2), medium (15 dyne/cm2) and high (30 dyne/cm2) via parallel plate flow chamber. RT-PCR was used to analyze the expression of MMP-9. The signaling inhibitors such as Wortmannin (PI3K specific inhabitor), SB202190 (p38MAPK specific inhabitor), and PD98059 (ERK1/2 specific inhabitor) were used to investigate the possible mechanical signal transduction pathway. The results showed: (1) The expression of MMP-9 was weak in static state, however, MMP-9 expression increased when MSCs were exposed to 15 dyne/cm2 shear stress for 2 hours, and MMP-9 expression increased with the extension of stimulating time, and it reached the peak at 24 h; (2) MSCs were stimulated by shear stress for 2 hours at different strengths (5 dyne/cm2, 15 dyne/cm2, 30 dyne/cm2), and under all these conditions, the expression of MMP-9 increased, and reached the peak at 15 dyne/cm2; (3) After MSCs were pretreated by three kinds of signal pathway inhibitors, the expression of MMP-9 did not change obviously in Wortmannin group and PD98059 group, but it was significantly inhibited in SB202190 group. This study demonstrated that shear stress could induce the expression of MMP-9 in rat bone marrow-derived mesenchymal stem cells; the amount of MMP-9 expression was closely related to stimulating time and the strengths of shear stress; and p38MAPK signal pathway played a critical role during the process.

  11. Cell multiplication, apoptosis and p-Akt protein expression of bone mesenchymal stem cells of rat under hypoxia environment

    Institute of Scientific and Technical Information of China (English)

    Hongliang Kong; Ningning Liu; Xin Huo; Bo Wang; Haipeng Zhang; Mingyu Gao; Guoxian Qi

    2007-01-01

    Objective :To elucidate whether cell multiplication, apoptosis, glucose intake and p-Akt protein expression of bone Mesenchymal Stem Cells(MSCs) of rats is influenced by a hypoxic environment ex vivo. Methods:Passage 3 of bone marrow MSCs taken from Wistar rats, were cultured in a culturing chamber with 94%N2,1%O2, 5%CO2 at 37℃. At different hypoxia time points, 0,0.5,1,4 and 8 h, glucose uptake was assayed by using radiation isotope 3H-G, Apoptotic Rate(AR) and dead rate(DR) were analyzed by flow cytometry(FCM) after Annexin V/PI staining, cell multiplication(by MTT methods) and p-Akt protein by immunocytochemistry and western blot. Results:Assay for CD29+,CD44+,CD71+,CD34-, Tn T+(after 5-azacytidine agent inducing) and ALP+(after bone differentiation agent inducing) suggested these bone-derived cells were MSCs. The 3H-G intaking ratio (CPM/flask value:157 ± 11,110 ± 11,107 ± 13,103 ± 10,100 ± 9 and 98 ± 10) of MSCs at different hypoxia time points, significantly decreased compared to that of normoxia(P < 0.01) and tended to descend slowly with hypoxia time duration, for which there was no statistical significance(P > 0.05). The AR(0.09 ± 2.03%,12.9 ± 1.72%,13.7 ± 2.26%,13.8 ± 3.01% ,14.1 ± 2.78% and 14.7 ±4.01% at 0,0.5,1,4 and 8 h,respectively,P < 0.01) and DR (0.04 ± 1.79% ,0.93 ± 1.85% ,3.11 ± 2.14%,4.09 ± 2.36% ,4.72 ±2.05% and 4.91 ± 3.72% at 0,0.5,1,4 and 8 h, respectively, P < 0.05) at different hypoxia time points significantly increased compared to those time in normoxia; The AR further went up with time (P < 0.05), however there was no statistical significance(P > 0.05) for the DR. Optical absorption value of MTT methods at different hypoxia time points significantly decreased compared to those with a corresponding normoxia time (P < 0.01 ) and degraded with time (in an hypoxic environment -P < 0.01 ).IOD of p-Akt protein of MSCs at different hypoxia time points significantly increased (0.367 ± 0.031,0.556 ± 0

  12. Reversal of hyperglycemia in diabetic rats by portal vein transplantation of islet-like cells generated from bone marrow mesenchymal stem cells

    Institute of Scientific and Technical Information of China (English)

    Xiao-Hong Wu; Chao Liu; Cui-Ping Liu; Kuan-Feng Xu; Xiao-Dong Mao; Jian Zhu; Jing-Jing Jiang; Dai Cui; Mei Zhang; Yu Xu

    2007-01-01

    AIM: To study the capacity of bone marrow mesenchymal stem cells (BM-MSCs) trans-differentiating into islet-like cells and to observe the effect of portal vein transplantation of islet-like cells in the treatment of streptozotocin-induced diabetic rat.METHODS: BM-MSCs were isolated from SD rats and induced to differentiate into islet-like cells under defined conditions. Differentiation was evaluated with electron microscopy, RT-PCR, immunofluorescence and flow cytometry. Insulin release after glucose challenge was tested with ELISA. Then allogeneic islet-like cells were transplanted into diabetic rats via portal vein. Blood glucose levels were monitored and islet hormones were detected in the liver and pancreas of the recipient by immunohistochemistry.RESULTS: BM-MSCs were spheroid adherent monolayers with high CD90, CD29 and very low CD45 expression.Typical islet-like cells clusters were formed after induction. Electron microscopy revealed that secretory granules were densely packed within the cytoplasm of the differentiated cells. The spheroid cells expressed islet related genes and hormones. The insulin-positive cells accounted for 19.8% and mean fluorescence intensity increased by 2.6 fold after induction. The cells secreted a small amount of insulin that was increased 1.5 fold after glucose challenge. After transplantation, islet-like cells could locate in the liver expressing islet hormones and lower the glucose levels of diabetic rats during d 6 to d 20.CONCLUSION: Rat BM-MSCs could be transdifferentiated into islet-like cells in vitro. Portal vein transplantation of islet-like cells could alleviate the hyperglycemia of diabetic rats.

  13. Ameliorative effects of human adipose tissue-derived mesenchymal stem cells on myelin basic protein-induced experimental autoimmune encephalomyelitis in Lewis rats

    Institute of Scientific and Technical Information of China (English)

    Myung-Soon Ko; Hyeong-geun Park; Young-Min Yun; Jeong Chan Ra; Taekyun Shin; Kyoung-Kap Lee

    2011-01-01

    Mesenchymal stem cells have been previously shown to exert an immunomodulatory function. The present study sought to investigate the effects of multipotential human adipose tissue-derived mesenchymal stem cells (hAdMSCs) on disease progression and cytokine expression in Lewis rats with experimental autoimmune encephalomyelitis (EAE) induced by myelin basic protein. The duration of EAE paralysis in the group treated on day 7 postimmunization with 5 × 106 hAdMSCs was significantly reduced compared with the vehicle-treated controls and the 1 × 106 hAdMSC- treated group. The duration of EAE paralysis in the groups treated with 5 × 106 hAdMSCs on both day 1 and day 7 postimmunization was significantly reduced compared with the vehicle-treated controls and the groups treated with 5 × 106 hAdMSCs on both day 7 and day 10 postimmunization. The mRNA expression of interleukin-10 and indoleamine 2, 3-dioxygenase was significantly decreased in the hAdMSC-treated group compared with the vehicle-treated group. These findings suggest that the ameliorative effects of hAdMSCs on EAE symptoms operate in a dose- and time-dependent manner and can be mediated in part by the ample production of anti-inflammatory cytokines.

  14. Deletion of a conserved regulatory element required for Hmx1 expression in craniofacial mesenchyme in the dumbo rat: a newly identified cause of congenital ear malformation

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    Lely A. Quina

    2012-11-01

    Hmx1 is a homeodomain transcription factor expressed in the developing eye, peripheral ganglia, and branchial arches of avian and mammalian embryos. Recent studies have identified a loss-of-function allele at the HMX1 locus as the causative mutation in the oculo-auricular syndrome (OAS in humans, characterized by ear and eye malformations. The mouse dumbo (dmbo mutation, with similar effects on ear and eye development, also results from a loss-of-function mutation in the Hmx1 gene. A recessive dmbo mutation causing ear malformation in rats has been mapped to the chromosomal region containing the Hmx1 gene, but the nature of the causative allele is unknown. Here we show that dumbo rats and mice exhibit similar neonatal ear and eye phenotypes. In midgestation embryos, dumbo rats show a specific loss of Hmx1 expression in neural-crest-derived craniofacial mesenchyme (CM, whereas Hmx1 is expressed normally in retinal progenitors, sensory ganglia and in CM, which is derived from mesoderm. High-throughput resequencing of 1 Mb of rat chromosome 14 from dmbo/dmbo rats, encompassing the Hmx1 locus, reveals numerous divergences from the rat genomic reference sequence, but no coding changes in Hmx1. Fine genetic mapping narrows the dmbo critical region to an interval of ∼410 kb immediately downstream of the Hmx1 transcription unit. Further sequence analysis of this region reveals a 5777-bp deletion located ∼80 kb downstream in dmbo/dmbo rats that is not apparent in 137 other rat strains. The dmbo deletion region contains a highly conserved domain of ∼500 bp, which is a candidate distal enhancer and which exhibits a similar relationship to Hmx genes in all vertebrate species for which data are available. We conclude that the rat dumbo phenotype is likely to result from loss of function of an ultraconserved enhancer specifically regulating Hmx1 expression in neural-crest-derived CM. Dysregulation of Hmx1 expression is thus a candidate mechanism for congenital ear

  15. Anti-Fibrosis Effect of Scutellarin via Inhibition of Endothelial–Mesenchymal Transition on Isoprenaline-Induced Myocardial Fibrosis in Rats

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    Hao Zhou

    2014-09-01

    Full Text Available Scutellarin (SCU is the major active component of breviscapine and has been reported to be capable of decreasing myocardial fibrosis. The aim of the present study is to investigate whether SCU treatment attenuates isoprenaline-induced myocardial fibrosis and the mechanisms of its action. Rats were injected subcutaneously with isoprenaline (Iso to induce myocardial fibrosis and rats in the SCU treatment groups were intraperitoneally infused with SCU (10 mg·kg−1·d−1 or 20 mg·kg−1·d−1, for 14 days. Post-treatment, cardiac functional measurements and the left and right ventricular weight indices (LVWI and RVWI, respectively were analysed. Pathological alteration, expression of type I and III collagen, Von Willebrand factor, α-smooth muscle actin, cluster of differentiation-31 (CD31, and the Notch signalling proteins (Notch1, Jagged1 and Hes1 were examined. The administration of SCU resulted in a significant improvement in cardiac function and decrease in the cardiac weight indices; reduced fibrous tissue proliferation; reduced levels of type I and III collagen; increased microvascular density; and decreased expression of α-smooth muscle actin and increased expression of CD31, Notch1, Jagged1 and Hes1 in isoprenaline-induced myocardial fibrosis in rats. Our results suggest that SCU prevents isoprenaline-induced myocardial fibrosis via inhibition of cardiac endothelial-mesenchymal transition potentially, which may be associated with the Notch pathway.

  16. Acellular spinal cord scaffold seeded with mesenchymal stem cells promotes long-distance axon regeneration and functional recovery in spinal cord injured rats.

    Science.gov (United States)

    Liu, Jia; Chen, Jian; Liu, Bin; Yang, Cuilan; Xie, Denghui; Zheng, Xiaochen; Xu, Song; Chen, Tianyu; Wang, Liang; Zhang, Zhongmin; Bai, Xiaochun; Jin, Dadi

    2013-02-15

    The stem cell-based experimental therapies are partially successful for the recovery of spinal cord injury (SCI). Recently, acellular spinal cord (ASC) scaffolds which mimic native extracellular matrix (ECM) have been successfully prepared. This study aimed at investigating whether the spinal cord lesion gap could be bridged by implantation of bionic-designed ASC scaffold alone and seeded with human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) respectively, and their effects on functional improvement. A laterally hemisected SCI lesion was performed in adult Sprague-Dawley (SD) rats (n=36) and ASC scaffolds seeded with or without hUCB-MSCs were implanted into the lesion immediately. All rats were behaviorally tested using the Basso-Beattie-Bresnahan (BBB) test once a week for 8weeks. Behavioral analysis showed that there was significant locomotor recovery improvement in combined treatment group (ASC scaffold and ASC scaffold+hUCB-MSCs) as compared with the SCI only group (pspinal cord cavity and promote long-distance axon regeneration and functional recovery in SCI rats.

  17. The Effect of Bone Marrow-Derived Mesenchymal Stem Cells and Their Conditioned Media Topically Delivered in Fibrin Glue on Chronic Wound Healing in Rats.

    Science.gov (United States)

    Mehanna, Radwa A; Nabil, Iman; Attia, Noha; Bary, Amany A; Razek, Khalid A; Ahmed, Tamer A E; Elsayed, Fatma

    2015-01-01

    Bone marrow-derived mesenchymal stem cells (BM-MSCs) represent a modern approach for management of chronic skin injuries. In this work, we describe BM-MSCs application versus their conditioned media (CM) when delivered topically admixed with fibrin glue to enhance the healing of chronic excisional wounds in rats. Fifty-two adult male rats were classified into four groups after induction of large-sized full-thickness skin wound: control group (CG), fibrin only group (FG), fibrin + MSCs group (FG + SCs), and fibrin + CM group (FG + CM). Healing wounds were evaluated functionally and microscopically. Eight days after injury, number of CD68+ macrophages infiltrating granulation tissue was considerably higher in the latter two groups. Although--later--none of the groups depicted a substantially different healing rate, the quality of regenerated skin was significantly boosted by the application of either BM-MSCs or their CM both (1) structurally as demonstrated by the obviously increased mean area percent of collagen fibers in Masson's trichrome-stained skin biopsies and (2) functionally as supported by the interestingly improved epidermal barrier as well as dermal tensile strength. Thus, we conclude that topically applied BM-MSCs and their CM-via fibrin vehicle--could effectively improve the quality of healed skin in chronic excisional wounds in rats, albeit without true acceleration of wound closure.

  18. Combination of low-energy shock-wave therapy and bone marrow mesenchymal stem cell transplantation to improve the erectile function of diabetic rats.

    Science.gov (United States)

    Shan, Hai-Tao; Zhang, Hai-Bo; Chen, Wen-Tao; Chen, Feng-Zhi; Wang, Tao; Luo, Jin-Tai; Yue, Min; Lin, Ji-Hong; Wei, An-Yang

    2017-01-01

    Stem cell transplantation and low-energy shock-wave therapy (LESWT) have emerged as potential and effective treatment protocols for diabetic erectile dysfunction. During the tracking of transplanted stem cells in diabetic erectile dysfunction models, the number of visible stem cells was rather low and decreased quickly. LESWT could recruit endogenous stem cells to the cavernous body and improve the microenvironment in diabetic cavernous tissue. Thus, we deduced that LESWT might benefit transplanted stem cell survival and improve the effects of stem cell transplantation. In this research, 42 streptozotocin-induced diabetic rats were randomized into four groups: the diabetic group (n = 6), the LESWT group (n = 6), the bone marrow-derived mesenchymal stem cell (BMSC) transplantation group (n = 15), and the combination of LESWT and BMSC transplantation group (n = 15). One and three days after BMSC transplantation, three rats were randomly chosen to observe the survival numbers of BMSCs in the cavernous body. Four weeks after BMSC transplantation, the following parameters were assessed: the surviving number of transplanted BMSCs in the cavernous tissue, erectile function, real-time polymerase chain reaction, and penile immunohistochemical assessment. Our research found that LESWT favored the survival of transplanted BMSCs in the cavernous body, which might be related to increased stromal cell-derived factor-1 expression and the enhancement of angiogenesis in the diabetic cavernous tissue. The combination of LESWT and BMSC transplantation could improve the erectile function of diabetic erectile function rats more effectively than LESWT or BMSC transplantation performed alone.

  19. The Effect of Bone Marrow-Derived Mesenchymal Stem Cells and Their Conditioned Media Topically Delivered in Fibrin Glue on Chronic Wound Healing in Rats

    Science.gov (United States)

    Mehanna, Radwa A.; Nabil, Iman; Attia, Noha; Bary, Amany A.; Razek, Khalid A.; Ahmed, Tamer A. E.; Elsayed, Fatma

    2015-01-01

    Bone marrow-derived mesenchymal stem cells (BM-MSCs) represent a modern approach for management of chronic skin injuries. In this work, we describe BM-MSCs application versus their conditioned media (CM) when delivered topically admixed with fibrin glue to enhance the healing of chronic excisional wounds in rats. Fifty-two adult male rats were classified into four groups after induction of large-sized full-thickness skin wound: control group (CG), fibrin only group (FG), fibrin + MSCs group (FG + SCs), and fibrin + CM group (FG + CM). Healing wounds were evaluated functionally and microscopically. Eight days after injury, number of CD68+ macrophages infiltrating granulation tissue was considerably higher in the latter two groups. Although—later—none of the groups depicted a substantially different healing rate, the quality of regenerated skin was significantly boosted by the application of either BM-MSCs or their CM both (1) structurally as demonstrated by the obviously increased mean area percent of collagen fibers in Masson's trichrome-stained skin biopsies and (2) functionally as supported by the interestingly improved epidermal barrier as well as dermal tensile strength. Thus, we conclude that topically applied BM-MSCs and their CM—via fibrin vehicle—could effectively improve the quality of healed skin in chronic excisional wounds in rats, albeit without true acceleration of wound closure. PMID:26236740

  20. The Effect of Bone Marrow-Derived Mesenchymal Stem Cells and Their Conditioned Media Topically Delivered in Fibrin Glue on Chronic Wound Healing in Rats

    Directory of Open Access Journals (Sweden)

    Radwa A. Mehanna

    2015-01-01

    Full Text Available Bone marrow-derived mesenchymal stem cells (BM-MSCs represent a modern approach for management of chronic skin injuries. In this work, we describe BM-MSCs application versus their conditioned media (CM when delivered topically admixed with fibrin glue to enhance the healing of chronic excisional wounds in rats. Fifty-two adult male rats were classified into four groups after induction of large-sized full-thickness skin wound: control group (CG, fibrin only group (FG, fibrin + MSCs group (FG + SCs, and fibrin + CM group (FG + CM. Healing wounds were evaluated functionally and microscopically. Eight days after injury, number of CD68+ macrophages infiltrating granulation tissue was considerably higher in the latter two groups. Although—later—none of the groups depicted a substantially different healing rate, the quality of regenerated skin was significantly boosted by the application of either BM-MSCs or their CM both (1 structurally as demonstrated by the obviously increased mean area percent of collagen fibers in Masson’s trichrome-stained skin biopsies and (2 functionally as supported by the interestingly improved epidermal barrier as well as dermal tensile strength. Thus, we conclude that topically applied BM-MSCs and their CM—via fibrin vehicle—could effectively improve the quality of healed skin in chronic excisional wounds in rats, albeit without true acceleration of wound closure.

  1. CdSe/ZnS Quantum Dots-Labeled Mesenchymal Stem Cells for Targeted Fluorescence Imaging of Pancreas Tissues and Therapy of Type 1 Diabetic Rats

    Science.gov (United States)

    Liu, Haoqi; Tang, Wei; Li, Chao; Lv, Pinlei; Wang, Zheng; Liu, Yanlei; Zhang, Cunlei; Bao, Yi; Chen, Haiyan; Meng, Xiangying; Song, Yan; Xia, Xiaoling; Pan, Fei; Cui, Daxiang; Shi, Yongquan

    2015-06-01

    Mesenchymal stem cells (MSCs) have been used for therapy of type 1 diabetes mellitus. However, the in vivo distribution and therapeutic effects of transplanted MSCs are not clarified well. Herein, we reported that CdSe/ZnS quantum dots-labeled MSCs were prepared for targeted fluorescence imaging and therapy of pancreas tissues in rat models with type 1 diabetes. CdSe/ZnS quantum dots were synthesized, their biocompatibility was evaluated, and then, the appropriate concentration of quantum dots was selected to label MSCs. CdSe/ZnS quantum dots-labeled MSCs were injected into mouse models with type 1 diabetes via tail vessel and then were observed by using the Bruker In-Vivo F PRO system, and the blood glucose levels were monitored for 8 weeks. Results showed that prepared CdSe/ZnS quantum dots owned good biocompatibility. Significant differences existed in distribution of quantum dots-labeled MSCs between normal control rats and diabetic rats ( p therapy of diabetic patients in the near future.

  2. Construction of the Eukaryotic Expression Vector with EGFP and hVE GF121 Gene and its Expression in Rat Mesenchymal Stem Cells

    Institute of Scientific and Technical Information of China (English)

    Su Li; Chen Yunzhen; Zhang Xiaogang; She Qiang

    2005-01-01

    Objectives To construct a recombinant plasmid carrying enhanced green fluorescent protein (EGFP) and human vascular endothelial growth factor (VEGF) 121 gene and detect its expression in rat mesenchymal stem cells (MSCs). Methods Human VEGF121 cDNA was amplified with polymerase chain reaction (PCR) from pCD/hVEGF121 and was inserted into the eukaryotic expression vector pEGFPC1. After being identified with PCR, double enzyme digestion and DNA sequencing. The recombinant plasmid pEGFP/hVEGF121 was transferred into rat MSCs with lipofectamine. The expression of EGFP/VEGF121 fusion protein were detected with fluorescence microscope and immunocytochemical staining respectively. Results The recombinant plasmid was confirmed with PCR, double enzyme digestion and DNA sequencing. The fluorescence microscope and immunocytochemical staining results showed that the EGFP and VEGF121 protein were expressed in MSCs 48 h after transfection.Conclusions The recombinant plasmid carrying EGFP and human VEGF was successfully constructed and expressed positively in rat MSCs. It offers a promise tool for further research on differentiation of MSCs and VEGF gene therapy for ischemial cardiovascular disease.

  3. Intravenous administration of mesenchymal stem cells exerts therapeutic effects on parkinsonian model of rats: Focusing on neuroprotective effects of stromal cell-derived factor-1α

    Directory of Open Access Journals (Sweden)

    Tayra Judith

    2010-04-01

    Full Text Available Abstract Background Mesenchymal stem cells (MSCs are pluripotent stem cells derived from bone marrow with secretory functions of various neurotrophic factors. Stromal cell-derived factor-1α (SDF-1α is also reported as one of chemokines released from MSCs. In this research, the therapeutic effects of MSCs through SDF-1α were explored. 6-hydroxydopamine (6-OHDA, 20 μg was injected into the right striatum of female SD rats with subsequent administration of GFP-labeled MSCs, fibroblasts, (i.v., 1 × 107 cells, respectively or PBS at 2 hours after 6-OHDA injection. All rats were evaluated behaviorally with cylinder test and amphetamine-induced rotation test for 1 month with consequent euthanasia for immunohistochemical evaluations. Additionally, to explore the underlying mechanisms, neuroprotective effects of SDF-1α were explored using 6-OHDA-exposed PC12 cells by using dopamine (DA assay and TdT-mediated dUTP-biotin nick-end labeling (TUNEL staining. Results Rats receiving MSC transplantation significantly ameliorated behaviorally both in cylinder test and amphetamine-induced rotation test compared with the control groups. Correspondingly, rats with MSCs displayed significant preservation in the density of tyrosine hydroxylase (TH-positive fibers in the striatum and the number of TH-positive neurons in the substantia nigra pars compacta (SNc compared to that of control rats. In the in vitro study, SDF-1α treatment increased DA release and suppressed cell death induced by 6-OHDA administration compared with the control groups. Conclusions Consequently, MSC transplantation might exert neuroprotection on 6-OHDA-exposed dopaminergic neurons at least partly through anti-apoptotic effects of SDF-1α. The results demonstrate the potentials of intravenous MSC administration for clinical applications, although further explorations are required.

  4. Impact of repeated intravenous bone marrow mesenchymal stem cells infusion on myocardial collagen network remodeling in a rat model of doxorubicin-induced dilated cardiomyopathy.

    Science.gov (United States)

    Yu, Qin; Li, Qianxiao; Na, Rongmei; Li, Xiaofei; Liu, Baiting; Meng, Lili; Liutong, Hanyu; Fang, Weiyi; Zhu, Ning; Zheng, Xiaoqun

    2014-02-01

    Bone marrow mesenchymal stem cells (MSCs) transplantation improved cardiac function and reduced myocardial fibrosis in both ischemic and non-ischemic cardiomyopathies. We evaluated the effects of repeated peripheral vein injection of MSCs on collagen network remodeling and myocardial TGF-β1, AT1, CYP11B2 (aldosterone synthase) gene expressions in a rat model of doxorubicin (DOX)-induced dilated cardiomyopathy (DCM). Thirty-eight out of 53 SD rats survived at 10 weeks post-DOX injection (2.5 mg/kg/week for 6 weeks, i.p.) were divided into DCM blank (without treatment, n = 12), DCM placebo (intravenous tail injection of 0.5 mL serum-free culture medium every other day for ten times, n = 13), and DCM plus MSCs group (intravenous tail injection of 5 × 10(6) MSCs dissolved in 0.5 mL serum-free culture medium every other day for 10 times, n = 13). Ten untreated rats served as normal controls. At 20 weeks after DOX injection, echocardiography, myocardial collagen content, myocardial expressions of types I and III collagen, TGF-β1, AT1, and CYP11B2 were compared among groups. At 20 weeks post-DOX injection, 8 rats (67%) survived in DCM blank group, 9 rats (69%) survived in DCM placebo group while 13 rats (100 %) survived in DCM plus MSCs group. Left ventricular end-diastolic diameter was significantly higher and ejection fraction was significantly lower in DCM blank and DCM placebo groups compared to normal control rats, which were significantly improved in DCM plus MSCs group (all p collagen volume fraction, types I and III collagen, myocardial mRNA expressions of TGF-β1, AT1, CYP11B2, and collagen I/III ratio were all significantly lower in DCM plus MSCs group compared to DCM blank and DCM placebo groups (all p collagen network remodeling possibly through downregulating renin-angiotensin-aldosterone system in DOX-induced DCM rats.

  5. The Therapeutic Potential of Human Umbilical Mesenchymal Stem Cells From Wharton's Jelly in the Treatment of Rat Peritoneal Dialysis-Induced Fibrosis.

    Science.gov (United States)

    Fan, Yu-Pei; Hsia, Ching-Chih; Tseng, Kuang-Wen; Liao, Chih-Kai; Fu, Tz-Win; Ko, Tsui-Ling; Chiu, Mei-Miao; Shih, Yang-Hsin; Huang, Pei-Yu; Chiang, Yi-Chia; Yang, Chih-Ching; Fu, Yu-Show

    2016-02-01

    A major complication in continuous, ambulatory peritoneal dialysis in patients with end-stage renal disease who are undergoing long-term peritoneal dialysis (PD) is peritoneal fibrosis, which can result in peritoneal structural changes and functional ultrafiltration failure. Human umbilical mesenchymal stem cells (HUMSCs) in Wharton's jelly possess stem cell properties and are easily obtained and processed. This study focuses on the effects of HUMSCs on peritoneal fibrosis in in vitro and in vivo experiments. After 24-hour treatment with mixture of Dulbecco's modified Eagle's medium and PD solution at a 1:3 ratio, primary human peritoneal mesothelial cells became susceptible to PD-induced cell death. Such cytotoxic effects were prevented by coculturing with primary HUMSCs. In a rat model, intraperitoneal injections of 20 mM methylglyoxal (MGO) in PD solution for 3 weeks (the PD/MGO 3W group) markedly induced abdominal cocoon formation, peritoneal thickening, and collagen accumulation. Immunohistochemical analyses indicated neoangiogenesis and significant increase in the numbers of ED-1- and α-smooth muscle actin (α-SMA)-positive cells in the thickened peritoneum in the PD/MGO 3W group, suggesting that PD/MGO induced an inflammatory response. Furthermore, PD/MGO treatment for 3 weeks caused functional impairments in the peritoneal membrane. However, in comparison with the PD/MGO group, intraperitoneal administration of HUMSCs into the rats significantly ameliorated the PD/MGO-induced abdominal cocoon formation, peritoneal fibrosis, inflammation, neoangiogenesis, and ultrafiltration failure. After 3 weeks of transplantation, surviving HUMSCs were found in the peritoneum in the HUMSC-grafted rats. Thus, xenografts of HUMSCs might provide a potential therapeutic strategy in the prevention of peritoneal fibrosis. Significance: This study demonstrated that direct intraperitoneal transplantation of human umbilical mesenchymal stem cells into the rat effectively

  6. Ultrasound-targeted stromal cell-derived factor-1-loaded microbubble destruction promotes mesenchymal stem cell homing to kidneys in diabetic nephropathy rats

    Directory of Open Access Journals (Sweden)

    Wu S

    2014-12-01

    Full Text Available Shengzheng Wu,1 Lu Li,1 Gong Wang,1 Weiwei Shen,2 Yali Xu,1 Zheng Liu,1 Zhongxiong Zhuo,1 Hongmei Xia,1 Yunhua Gao,1 Kaibin Tan1 1Department of Ultrasound, 2Department of Orthopedics, Xinqiao Hospital, Third Military Medical University, Chongqing, People’s Republic of China Abstract: Mesenchymal stem cell (MSC therapy has been considered a promising strategy to cure diabetic nephropathy (DN. However, insufficient MSCs can settle in injured kidneys, which constitute one of the major barriers to the effective implementation of MSC therapy. Stromal cell-derived factor-1 (SDF-1 plays a vital role in MSC migration and involves activation, mobilization, homing, and retention, which are presumably related to the poor homing in DN therapy. Ultrasound-targeted microbubble destruction has become one of the most promising strategies for the targeted delivery of drugs and genes. To improve MSC homing to DN kidneys, we present a strategy to increase SDF-1 via ultrasound-targeted microbubble destruction. In this study, we developed SDF-1-loaded microbubbles (MBSDF-1 via covalent conjugation. The characterization and bioactivity of MBSDF-1 were assessed in vitro. Target release in the targeted kidneys was triggered with diagnostic ultrasound in combination with MBSDF-1. The related bioeffects were also elucidated. Early DN was induced in rats with streptozotocin. Green fluorescent protein-labeled MSCs were transplanted intravenously following the target release of SDF-1 in the kidneys of normal and DN rats. The homing efficacy was assessed by detecting the implanted exogenous MSCs at 24 hours. The in vitro results showed an impressive SDF-1 loading efficacy of 79% and a loading content of 15.8 µg/mL. MBSDF-1 remained bioactive as a chemoattractant. In the in vivo study, SDF-1 was successfully released in the targeted kidneys. The homing efficacy of MSCs to DN kidneys after the target release of SDF-1 was remarkably ameliorated at 24 hours compared with

  7. MicroRNA-9 promotes the neuronal differentiation of rat bone marrow mesenchymal stem cells by activating autophagy

    Directory of Open Access Journals (Sweden)

    Guang-yu Zhang

    2015-01-01

    Full Text Available MicroRNA-9 (miR-9 has been shown to promote the differentiation of bone marrow mesenchymal stem cells into neuronal cells, but the precise mechanism is unclear. Our previous study confirmed that increased autophagic activity improved the efficiency of neuronal differentiation in bone marrow mesenchymal stem cells. Accumulating evidence reveals that miRNAs adjust the autophagic pathways. This study used miR-9-1 lentiviral vector and miR-9-1 inhibitor to modulate the expression level of miR-9. Autophagic activity and neuronal differentiation were measured by the number of light chain-3 (LC3-positive dots, the ratio of LC3-II/LC3, and the expression levels of the neuronal markers enolase and microtubule-associated protein 2. Results showed that LC3-positive dots, the ratio of LC3-II/LC3, and expression of neuron specific enolase and microtubule-associated protein 2 increased in the miR-9 + group. The above results suggest that autophagic activity increased and bone marrow mesenchymal stem cells were prone to differentiate into neuronal cells when miR-9 was overexpressed, demonstrating that miR-9 can promote neuronal differentiation by increasing autophagic activity.

  8. MicroRNA-9 promotes the neuronal differentiation of rat bone marrow mesenchymal stem cells by activating autophagy

    Institute of Scientific and Technical Information of China (English)

    Guang-yu Zhang; Jun Wang; Yan-jie Jia; Rui Han; Ping Li; Deng-na Zhu

    2015-01-01

    MicroRNA-9 (miR-9) has been shown to promote the differentiation of bone marrow mesen-chymal stem cells into neuronal cells, but the precise mechanism is unclear. Our previous study conifrmed that increased autophagic activity improved the efifciency of neuronal differentiation in bone marrow mesenchymal stem cells. Accumulating evidence reveals that miRNAs adjust the autophagic pathways. This study used miR-9-1 lentiviral vector and miR-9-1 inhibitor to modulate the expression level of miR-9. Autophagic activity and neuronal differentiation were measured by the number of light chain-3 (LC3)-positive dots, the ratio of LC3-II/LC3, and the expression levels of the neuronal markers enolase and microtubule-associated protein 2. Re-sults showed that LC3-positive dots, the ratio of LC3-II/LC3, and expression of neuron speciifc enolase and microtubule-associated protein 2 increased in the miR-9+ group. The above results suggest that autophagic activity increased and bone marrow mesenchymal stem cells were prone to differentiate into neuronal cells when miR-9 was overexpressed, demonstrating that miR-9 can promote neuronal differentiation by increasing autophagic activity.

  9. Ultrasound-targeted stromal cell-derived factor-1-loaded microbubble destruction promotes mesenchymal stem cell homing to kidneys in diabetic nephropathy rats.

    Science.gov (United States)

    Wu, Shengzheng; Li, Lu; Wang, Gong; Shen, Weiwei; Xu, Yali; Liu, Zheng; Zhuo, Zhongxiong; Xia, Hongmei; Gao, Yunhua; Tan, Kaibin

    2014-01-01

    Mesenchymal stem cell (MSC) therapy has been considered a promising strategy to cure diabetic nephropathy (DN). However, insufficient MSCs can settle in injured kidneys, which constitute one of the major barriers to the effective implementation of MSC therapy. Stromal cell-derived factor-1 (SDF-1) plays a vital role in MSC migration and involves activation, mobilization, homing, and retention, which are presumably related to the poor homing in DN therapy. Ultrasound-targeted microbubble destruction has become one of the most promising strategies for the targeted delivery of drugs and genes. To improve MSC homing to DN kidneys, we present a strategy to increase SDF-1 via ultrasound-targeted microbubble destruction. In this study, we developed SDF-1-loaded microbubbles (MB(SDF-1)) via covalent conjugation. The characterization and bioactivity of MB(SDF-1) were assessed in vitro. Target release in the targeted kidneys was triggered with diagnostic ultrasound in combination with MB(SDF-1). The related bioeffects were also elucidated. Early DN was induced in rats with streptozotocin. Green fluorescent protein-labeled MSCs were transplanted intravenously following the target release of SDF-1 in the kidneys of normal and DN rats. The homing efficacy was assessed by detecting the implanted exogenous MSCs at 24 hours. The in vitro results showed an impressive SDF-1 loading efficacy of 79% and a loading content of 15.8 μg/mL. MB(SDF-1) remained bioactive as a chemoattractant. In the in vivo study, SDF-1 was successfully released in the targeted kidneys. The homing efficacy of MSCs to DN kidneys after the target release of SDF-1 was remarkably ameliorated at 24 hours compared with control treatments in normal rats and DN rats. In conclusion, ultrasound-targeted MB(SDF-1) destruction could promote the homing of MSCs to early DN kidneys and provide a novel potential therapeutic approach for DN kidney repair.

  10. Long Term Study of Protective Mechanisms of Human Adipose Derived Mesenchymal Stem Cells on Cisplatin Induced Kidney injury in Sprague-Daweley Rats

    Directory of Open Access Journals (Sweden)

    Elhusseini FM

    2016-05-01

    Full Text Available Background/Aims: Long-term evaluation of cisplatin induced nephrotoxicity and the probable renal protective activities of stem cells are lacking up until now. We evaluated the early and long-term role of human adipose derived mesenchymal stem cells (ADMSCs in prevention or amelioration of cisplatin induced acute kidney injury (AKI in Sprague-Dawley rats. For this, we determined the kidney tissue level of oxidative stress markers in conjugation with a renal histopathological scoring system of both acute and chronic renal changes. Methods: This study used eighty Sprague-Dawley (SD rats weighing 250-300g. They were assigned into four equal groups (each group n=20: (I Negative control group, rats injected with single dose of 1 ml normal saline. (II Positive control cisplatin, rats injected with a single dose of 5 mg/kg I.P in 1 ml saline. (III Cisplatin and culture media group, rats injected with 0.5 ml of culture media single dose into the tail vein and (IV Cisplatin and ADMSCs group, rats injected with a single dose of 0.5 ml of culture media containing 5 x106ADMSCs into the tail vein one day after cisplatin administration. Each main group was further divided according to the timing of sacrifice into four subgroups (each subgroup n=5. Rats in the subgroup A were sacrificed after 4 days; subgroup B were sacrificed after 7 days; subgroup C were sacrificed after 11 days; and subgroup D were sacrificed after 30 days. Before sacrifice, 24 hrs.-urine was collected using a metabolic cage. Renal function was evaluated through blood urea nitrogen (BUN, serum creatinine and creatinine clearance. Kidney tissue homogenate oxidative stress parameters, Malondialdehyde (MDA, Superoxide dismutase (SOD and Glutathione (GSH were determined. In addition, histopathological analysis for active injury, regenerative and chronic changes was performed. Results: ADMSCs were characterized and their capability of differentiation was proved. Cisplatin induced a significant

  11. A model for chemically-induced mechanical loading on MEMS

    DEFF Research Database (Denmark)

    Amiot, Fabien

    2007-01-01

    The development of full displacement field measurements as an alternative to the optical lever technique to measure the mechanical response for microelectro-mechanical systems components in their environment calls for a modeling of chemically-induced mechanical fields (stress, strain, and displac......The development of full displacement field measurements as an alternative to the optical lever technique to measure the mechanical response for microelectro-mechanical systems components in their environment calls for a modeling of chemically-induced mechanical fields (stress, strain...

  12. Effects of lateral ventricular transplantation of bone marrow-derived mesenchymal stem cells modified with brain-derived neurotrophic factor gene on cognition in a rat model of Alzheimer's disease

    Institute of Scientific and Technical Information of China (English)

    Ping Zhang; Gangyong Zhao; Xianjiang Kang; Likai Su

    2012-01-01

    In the present study, transplantation of bone marrow-derived mesenchymal stem cells modified with brain-derived neurotrophic factor gene into the lateral ventricle of a rat model of Alzheimer's disease, resulted in significant attenuation of nerve cell damage in the hippocampal CA1 region. Furthermore, brain-derived neurotrophic factor and tyrosine kinase B mRNA and protein levels were significantly increased, and learning and memory were significantly improved. Results indicate that transplantation of bone marrow-derived mesenchymal stem cells modified with brain-derived neurotrophic factor gene can significantly improve cognitive function in a rat model of Alzheimer's disease, possibly by increasing the levels of brain-derived neurotrophic factor and tyrosine kinase B in the hippocampus.

  13. Efficacy of Atorvastatin combined with adipose-derived mesenchymal stem cell transplantation on cardiac function in rats with acute myocardial infarction

    Institute of Scientific and Technical Information of China (English)

    Anping Cai; Jian Kuang; Gang Dai; Weiyi Mai; Dongdan Zheng; Yugang Dong; Ruofeng Qiu; Yuli Huang; Yuanbin Song; Zhigao Jiang; Shaoqi Rao; Xinxue Liao

    2011-01-01

    Mesenchymal stem cells (MSCs) have been extensively applied for the restoration of cardiomyocytes loss after acute myocardial infarction (AMI).However,the optimal therapeutic efficacy of MSCs in ischemic heart diseases has been hampered by their poor survival and low differentiated rates.Therefore,the improvement of MSC survival and differentiated rates is warranted and critical for the efficacy of MSCs in AMI.In this paper,MSCs isolated from rat inguinal fat tissues were termed as adiposederived mesenchymal stem cells (ASCs),and the fourth passage of ASCs was pre-specified by co-culturing with cardiomyocytes in a transwell system termed as co-ASCs.Fourteen days later,GATA-4 (a transcription factor) and cardiac troponin-Ⅰ were detected by cellular immunofluorescence.Atorvastatin (Ator group) or vehicle (control group) was administrated for the first 24 h after AMI production in rats.Fourteen days later,inflammatory parameters and cardiac function were evaluated.The other surviving rats were injected with a total of 1 × 106 co-ASCs/100 μ1 phosphate-buffered saline (PBS),1 × 106 ASCs/100 μl PBS,or 100 μl PBS.Twenty-eight days after cell injection,survival and differentiated rates of transplanted cells and cardiac function were evaluated.The percentage of GATA-4 expression in co-ASCs was 28.5% ± 5.6% and of cardiac troponin-Ⅰ was 22.8% ±3.2%.Compared with the control group,the number of infiltrating inflammatory cells,myeloperoxidase activity,inflammatory cytokines (VCAM-1, TNF-α, Hs-CRP)mRNA expression,and Bax protein expression were significantly reduced in the three Ator groups,accompanied by a significant improvement of Bcl-2 protein expression and cardiac function (P<0.05).Compared with the Ator2 + ASCs group and Con + co-ASCs group,the number of 4-6-diamidino-2-phenylindole-stained cells and cardiac troponin-I-positive transplanted cells,concomitant with cardiac function,were improved most prominently in the Ator3 + co-ASCs group (P<0

  14. The effect of mesenchymal stem cell transplantation on the recovery of bladder and hindlimb function after spinal cord contusion in rats

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    Park Jeong-Soo

    2010-09-01

    Full Text Available Abstract Background Mesenchymal stem cells are widely used for transplantation into the injured spinal cord in vivo model and for safety, many human clinical trials are continuing to promote improvements of motor and sensory functions after spinal cord injury. Yet the exact mechanism for these improvements remains undefined. Neurogenic bladder following spinal cord injury is the main problem decreasing the quality of life for patients with spinal cord injury, but there are no clear data using stem cell transplantation for the improvement of neurogenic bladder for in vivo studies and the clinical setting. The purpose of this study was to delineate the effect of human mesenchymal stem cell (hMSCs transplantation on the restoration of neurogenic bladder and impaired hindlimb function after spinal cord contusion of rats and the relationship between neurotrophic factors such as brain derived neurotrophic factor (BDNF and neurotrophin-3 (NT-3 and bladder and hindlimb functions. Results Modified moderate contusion injury were performed on the thoracic spinal cord of Sprague-Dawley rats using MASCIS impactor and hMSCs, human fibroblasts or phosphate-buffered saline were transplanted into injured spinal cord 9 days after injury for hMSC and two control groups respectively. Ladder test showed more rapid restoration of hindlimb function in hMSC group than in control group, but Basso, Beattie, and Bresnahan score and coupling score were not different significantly among hMSC and two control groups. Neurogenic bladder was not improved in either group. ED1 positive macrophages were significantly reduced in hMSC group than in two control groups, but ELISA and RT-PCR studies revealed BDNF and NT-3 levels in spinal cord and bladder were not different among hMSC and two control groups regardless the experimental duration. Conclusion hMSC transplantation was effective in reducing inflammatory reaction after spinal cord contusion of rats but not sufficient to

  15. Effect of Transplantation of Bone Marrow Derived Mesenchymal Stem Cells and Platelets Rich Plasma on Experimental Model of Radiation Induced Oral Mucosal Injury in Albino Rats

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    Basma Elsaadany

    2017-01-01

    Full Text Available Normal tissue damage following radiotherapy is still a major problem in cancer treatment. Therefore, the current work aimed at exploring the possible role of systemically injected bone marrow derived mesenchymal stem cells (BM-MSCs and/or locally injected platelet rich plasma (PRP in ameliorating the side effects of ionizing radiation on the rat’s tongue. Twelve rats served as control group (N and 48 rats received a single radiation dose of 13 Gy to the head and neck region; then, they were equally divided into 4 experimental groups: irradiated only (C, irradiated + MSCs (S, irradiated + (PRP (P, and combined group (PS. Animal scarification occurred in 3 and 7 days after radiation. Then, tongues were dissected and examined histologically and for expression of bcl-2 by RT-PCR. Histological examination of the treated groups (S, (P, and (PS revealed an obvious improvement in the histological structure of the tongue, compared to group (C, in addition to upregulated expression of bcl-2, indicating decreased apoptotic activity. Conclusion. BM-MSCs and PRP have shown positive effect in minimizing the epithelial atrophy of normal oral mucosa after regional radiotherapy, which was emphasized by decreasing apoptotic activity in these tissues. Nevertheless, combined use of BM-MSCs and PRP did not reveal the assumed synergetic effect in oral tissue protection.

  16. Microbubble-mediated ultrasound promotes accumulation of bone marrow mesenchymal stem cell to the prostate for treating chronic bacterial prostatitis in rats.

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    Yi, Shanhong; Han, Guangwei; Shang, Yonggang; Liu, Chengcheng; Cui, Dong; Yu, Shuangjiang; Liao, Bin; Ao, Xiang; Li, Guangzhi; Li, Longkun

    2016-01-22

    Chronic bacterial prostatitis (CBP) is an intractable disease. Although bone marrow mesenchymal stem cells (BMMSCs) are able to regulate inflammation in CBP, the effect of microbubble-mediated ultrasound- induced accumulation of BMMSCs on CBP remains unclear. To address this gap, a model of CBP was established in SD rats, which were then treated with BMMSCs alone (BMMSC group), BMMSCs with ultrasound (ultrasound group), BMMSCs with microbubble-mediated ultrasound (MMUS group) and compared with a healthy control group. A therapeutic-ultrasound apparatus was used to treat the prostate in the presence of circulating microbubbles and BMMSCs. The BMMSC distribution was assessed with in vivo imaging, and the prostate structure with light microscopy. Real-time quantitative RT-PCR, ELISA, and immunohistochemistry were used to assess the expressions of TNF-α and IL-1β. More BMMSCs were found in the prostate in the MMUS group than in the CBP, ultrasound, and BMMSC groups. Inflammatory cell infiltration, fibrous tissue hyperplasia, and tumor-like epithelial proliferation were significantly reduced in the MMUS group, as were the mRNA and protein expressions of TNF-α and IL-1β. Microbubble-mediated ultrasound-induced accumulation of BMMSCs can inhibit inflammation and decrease TNF-α and IL-1β expressions in the prostate of CBP rats, suggesting that this method may be therapeutic for CPB.

  17. Human umbilical cord blood-derived mesenchymal stem cells do not differentiate into neural cell types or integrate into the retina after intravitreal grafting in neonatal rats.

    Science.gov (United States)

    Hill, Andrew J; Zwart, Isabel; Tam, Henry H; Chan, Jane; Navarrete, Cristina; Jen, Ling-Sun; Navarrete, Roberto

    2009-04-01

    This study investigated the ability of mesenchymal stem cells (MSCs) derived from full-term human umbilical cord blood to survive, integrate and differentiate after intravitreal grafting to the degenerating neonatal rat retina following intracranial optic tract lesion. MSCs survived for 1 week in the absence of immunosuppression. When host animals were treated with cyclosporin A and dexamethasone to suppress inflammatory and immune responses, donor cells survived for at least 3 weeks, and were able to spread and cover the entire vitreal surface of the host retina. However, MSCs did not significantly integrate into or migrate through the retina. They also maintained their human antigenicity, and no indication of neural differentiation was observed in retinas where retinal ganglion cells either underwent severe degeneration or were lost. These results have provided the first in vivo evidence that MSCs derived from human umbilical cord blood can survive for a significant period of time when the host rat response is suppressed even for a short period. These results, together with the observation of a lack of neuronal differentiation and integration of MSCs after intravitreal grafting, has raised an important question as to the potential use of MSCs for neural repair through the replacement of lost neurons in the mammalian retina and central nervous system.

  18. Stimulatory effect of HGF-overexpressing adipose tissue-derived mesenchymal stem cells on thymus regeneration in a rat thymus involution model.

    Science.gov (United States)

    Jung, Woo-Sung; Han, Sei-Myoung; Kim, Sung-Min; Kim, Mi-Eun; Lee, Jun-Sik; Seo, Kyoung-Won; Youn, Hwa-Young; Lee, Hee-Woo

    2014-10-01

    The thymus is the central lymphoid organ providing a unique and essential microenvironment for T-cell precursor development into mature functionally competent T-lymphocytes. Thus, it is important to develop the strategies for enhancing thymic regeneration from involution induced by a variety of clinical treatments and conditions. Hepatocyte growth factor (HGF) promotes proliferation in a variety of cell types. We have used stem cell-based HGF gene therapy to enhance regeneration from acute thymic involution. HGF-overexpressing human adipose tissue-derived mesenchymal stem cells (HGF-hATMSCs) were generated by liposomal transfection with the pMEX expression vector, constructed by inserting the HGF gene. Significantly increased HGF expression in these cells was confirmed by reverse transcription-polymerase chain reaction and an enzyme-linked immunosorbent assay. HGF produced by HGF-hATMSCs enhanced the proliferation of a mouse thymic epithelial cell line and the expression of interleukin-7 in vitro. We also examined the effect of HGF-hATMSCs on thymic regeneration in rats with acute thymic involution. Significant increases in thymus size and weight, as well as the number of thymocytes (especially, early thymocyte progenitors), were seen in the HGF-hATMSCs-treated rats compared to saline-treated control animals. A stimulatory effect of HGF-hATMSCs on thymic regeneration has therefore been shown, highlighting the clinical value of HGF-hATMSCs for treating thymic involution.

  19. Application potential of bone marrow mesenchymal stem cell (BMSCs) based tissue-engineering for spinal cord defect repair in rat fetuses with spina bifida aperta.

    Science.gov (United States)

    Li, Xiaoshuai; Yuan, Zhengwei; Wei, Xiaowei; Li, Hui; Zhao, Guifeng; Miao, Jiaoning; Wu, Di; Liu, Bo; Cao, Songying; An, Dong; Ma, Wei; Zhang, Henan; Wang, Weilin; Wang, Qiushi; Gu, Hui

    2016-04-01

    Spina bifida aperta are complex congenital malformations resulting from failure of fusion in the spinal neural tube during embryogenesis. Despite surgical repair of the defect, most patients who survive with spina bifida aperta have a multiple system handicap due to neuron deficiency of the defective spinal cord. Tissue engineering has emerged as a novel treatment for replacement of lost tissue. This study evaluated the prenatal surgical approach of transplanting a chitosan-gelatin scaffold seeded with bone marrow mesenchymal stem cells (BMSCs) in the healing the defective spinal cord of rat fetuses with retinoic acid induced spina bifida aperta. Scaffold characterisation revealed the porous structure, organic and amorphous content. This biomaterial promoted the adhesion, spreading and in vitro viability of the BMSCs. After transplantation of the scaffold combined with BMSCs, the defective region of spinal cord in rat fetuses with spina bifida aperta at E20 decreased obviously under stereomicroscopy, and the skin defect almost closed in many fetuses. The transplanted BMSCs in chitosan-gelatin scaffold survived, grew and expressed markers of neural stem cells and neurons in the defective spinal cord. In addition, the biomaterial presented high biocompatibility and slow biodegradation in vivo. In conclusion, prenatal transplantation of the scaffold combined with BMSCs could treat spinal cord defect in fetuses with spina bifida aperta by the regeneration of neurons and repairmen of defective region.

  20. In vivo tracing of superparamagnetic iron oxide-labeled bone marrow mesenchymal stem cells transplanted for traumatic brain injury by susceptibility weighted imaging in a rat model

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    CHENG Jing-liang; YANG Yun-jun; LI Hua-li; WANG Juan; WANG Mei-hao; ZHANG Yong

    2010-01-01

    Objective:To label rat bone marrow mesenchymal stem cells (BMSCs) with superparamagnetic iron oxide (SPIO) in vitro, and to monitor the survival and location of these labeled BMSCs in a rat model of traumatic brain injury (TBI) by susceptibility weighted imaging (SWI)sequence.Methods:BMSCs were cultured in vitro and then labeled with SPIO. Totally 24 male Sprague Dawley (SD) rats weighing 200-250 g were randomly divided into 4 groups: Groups A-D (n=6 for each group). Moderate TBI models of all the rats were developed in the left hemisphere following Feeney's method. Group A was the experimental group and stereotaxic transplantation of BMSCs labeled with SPIO into the region nearby the contusion was conducted in this group 24 hours after TBI modeling. The other three groups were control groups with transplantation of SPIO, unlabeled BMSCs and injection of nutrient solution respectively conducted in Groups B, C and D at the same time. Monitoring of these SPIO-labeled BMSCs by SWI was performed one day,one week and three weeks after implantation.Results: Numerous BMSCs were successfully labeled with SPIO. They were positive for Prussian blue staining and intracytoplasm positive blue stained particles were found under a microscope (×200). Scattered little iron particles were observed in the vesicles by electron microscopy (×5000). MRI of the transplantation sites of the left hemisphere demonstrated a low signal intensity on magnitude images,phase images and SWI images for all the test rats in Group A, and the lesion in the left parietal cortex demonstrated a semicircular low intensity on SWI images, which clearly showed the distribution and migration of BMSCs in the first and third weeks. For Group B, a low signal intensity by MRI was only observed on the first day but undetected during the following examination. No signals were observed in Groups C and D at any time points.Conclusion:SWI sequence in vivo can consecutively and noninvasively trace and demonstrate the

  1. The Chemopreventive Effect of Tamoxifen Combined with Celecoxib on DMBA chemically-Induced Breast Cancer

    Institute of Scientific and Technical Information of China (English)

    Xiaoxu Liu; Huafeng Kang; Xijing Wang; Zhijun Dai; Fengjie Xue; Xinghuan Xue

    2007-01-01

    Objective: To investigate the chemopreventive effect of tamoxifen combined with a COX-2 selective inhibitor, celecoxib, on breast cancer in rats chemically induced by 7,12-dimethylben (a)anthracene (DMBA). Methods:DMBA was irrigated into the stomaches of SD female rats to build breast cancer model. A total of 120 rats were divided into four groups: control group, tamoxifen group, celecoxib group and combined group. The incidence rate, latent period, number and volume of breast cancer were detected and analyzed. Results:The tumor incidence rate of tamoxifen group (48.15%, 13/27) and celecoxib group (50.00%,14/28) were lower than that of control group (85.71%, 24/28), but higher than that of combined group (21.43%, 6/28). The tumor's latent period of tamoxifen group (97.54±1.85 d) and celecoxib group (96.79±2.89 d) were longer than that of control group (89.50±5.99 d), but shorter than that of combined group (103.67±3.39 d). The average tumor number of tamoxifen group (1.77±0.73) and celecoxib group (1.71±0.61) were less than that of control group (3.50±1.62), but more than that of combined group ( 1.17±0.42 ). The average tumor volume of tamoxifen group (1.78±0.71 cm3) and celecoxib group (2.05±1.04 cm3) were smaller than that of control group (6.42±3.96 cm3), but bigger than that of combined group (0.71±0.96 cm3) (P < 0.05 respectively).Conclusion:Celecoxib and tamoxifen are effective drugs in preventing the occurrence of rat breast cancer chemically induced by DMBA. Furthermore, combination of them has better chemopreventive effect.

  2. Transplants of adult mesenchymal and neural stem cells provide neuroprotection and behavioral sparing in a transgenic rat model of Huntington's disease.

    Science.gov (United States)

    Rossignol, Julien; Fink, Kyle; Davis, Kendra; Clerc, Steven; Crane, Andrew; Matchynski, Jessica; Lowrance, Steven; Bombard, Matthew; Dekorver, Nicholas; Lescaudron, Laurent; Dunbar, Gary L

    2014-02-01

    Stem cells have gained significant interest as a potential treatment of neurodegenerative diseases, including Huntington's disease (HD). One source of these cells is adult neural stem cells (aNSCs), which differentiate easily into neuronal lineages. However, these cells are vulnerable to immune responses following transplantation. Another source is bone-marrow-derived mesenchymal stem cells (MSCs), which release neurotrophic factors and anti-inflammatory cytokines following transplantation, and are less vulnerable to rejection. The goal of this study was to compare the efficacy of transplants of MSCs, aNSCs, or cotransplants of MSCs and aNSCs for reducing deficits in a transgenic rat model of HD. HD rats received intrastriatal transplantations of 400,000 MSCs, aNSCs, or a combination of MSCs/aNSCs, while wild-type and HD controls were given vehicle. Rats were tested on the rotarod over the course of 20 weeks. The results indicated that transplants of: (a) aNSCs produced a strong immune response and conferred short-term behavioral benefits; (b) MSCs elicited a relatively weak immune response, and provided a longer term behavioral benefit; and (c) combined MSCs and aNSCs conferred long-term behavioral benefits and increased survival of the transplanted aNSCs. The finding that cotransplanting MSCs with aNSCs can prolong aNSC survival and provide greater behavioral sparing than when the transplants contains only aNSCs suggests that MSCs are capable of creating a more suitable microenvironment for aNSC survival. This cotransplantation strategy may be useful as a future therapeutic option for treating HD, especially if long-term survival of differentiated cells proves to be critically important for preserving lasting functional outcomes.

  3. Effect of transplanted mesenchymal stem cells from rats of different ages on the improvement of heart function after acute myocardial infarction

    Institute of Scientific and Technical Information of China (English)

    WANG Yi-qing; WANG Miao; ZHANG Peng; SONG Jing-jin; LI Yuan-peng; HOU Shu-hong; HUANG Cong-xin

    2008-01-01

    Background Mesenchymal stem cells (MSCs) transplantation is of therapeutic potential after ischemic injury in both experimental and clinical studies.Clinically,elderly patients are more vulnerable to acute myocardial infarction (AMI).But little is known about the characteristics of young donor-derived MSCs transplanted to old patients with AMI.The present study was designed to investigate the effect of transplanted MSCs from rats of different ages on the improvement of heart function after AMI.Methods MSCs from Sprague-Dawley (SD) rats were isolated and cultured in vitro.The apoptosis characteristics of MSCs were observed under conditions of ischemia and anoxia.SD rats underwent MI received intramyocardial injection of MSCs from young donor rats (n=8),old donor rats (n=8),respectively.AMI control group received equal volume physiological saline.Immunofluorescence was used to observe the differentiation of the grafted cells into cardiomyocytes.Four weeks after cell transplantation, reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry for vascular endothelial growth factor (VEGF),VIII-factor immunohistochemistry for vessel density,TUNEL,caspase-3 for cardiomyocyte apoptosis,echocardiography and hemodynamic detection for heart function were performed.Results The apoptosis rate of the old donor-derived MSCs group was significantly higher than that of the young donor-derived MSCs group under conditions of ischemia and anoxia (P <0.05).Engrafted MSCs survived,proliferated and differentiated into myocardium-like cells.VEGF gene expression and capillary density in the old donor-derived group were lower than those in the young donor-derived group but higher than those in the control group (P <0.05).The transplantation of old donor-derived MSCs attenuated apoptosis of cadiomyocytes in the peri-infract region compared with the control group and the effect was elevated in young donor-derived MSCs (P <0.05).The heart functions (left ventricle

  4. Biofunctionalization of a titanium surface with a nano-sawtooth structure regulates the behavior of rat bone marrow mesenchymal stem cells

    Directory of Open Access Journals (Sweden)

    Zhang WJ

    2012-08-01

    Full Text Available Wenjie Zhang,1,2 Zihui Li,3 Yan Liu,1,2 Dongxia Ye,4 Jinhua Li,3 Lianyi Xu,1,2 Bin Wei,1 Xiuli Zhang,2 Xuanyong Liu,3,* Xinquan Jiang,1,2,* 1Department of Prosthodontics, Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, 2Oral Bioengineering Laboratory, Shanghai Research Institute of Stomatology, Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, Shanghai Key Laboratory of Stomatology, 3State Key Laboratory of High Performance Ceramics and Superfine Microstructure, Shanghai Institute of Ceramics, Chinese Academy of Sciences, 4Shanghai Research Institute of Stomatology, Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, Shanghai Key Laboratory of Stomatology, Shanghai, China*Joint principal authors of this workBackground: The topography of an implant surface can serve as a powerful signaling cue for attached cells and can enhance the quality of osseointegration. A series of improved implant surfaces functionalized with nanoscale structures have been fabricated using various methods. Methods: In this study, using an H2O2 process, we fabricated two size-controllable sawtooth-like nanostructures with different dimensions on a titanium surface. The effects of the two nano-sawtooth structures on rat bone marrow mesenchymal stem cells (BMMSCs were evaluated without the addition of osteoinductive chemical factors.Results: These new surface modifications did not adversely affect cell viability, and rat BMMSCs demonstrated a greater increase in proliferation ability on the surfaces of the nano-sawtooth structures than on a control plate. Furthermore, upregulated expression of osteogenic-related genes and proteins indicated that the nano-sawtooth structures promote osteoblastic differentiation of rat BMMSCs. Importantly, the large nano-sawtooth structure resulted in the greatest cell responses, including increased adhesion, proliferation

  5. Upregulation of CC Chemokine Receptor 7 (CCR7) Enables Migration of Xenogeneic Human Adipose-Derived Mesenchymal Stem Cells to Rat Secondary Lymphoid Organs.

    Science.gov (United States)

    Ma, Tian; Luan, Shao-Liang; Huang, Hong; Sun, Xing-Kun; Yang, Yan-Mei; Zhang, Hui; Han, Wei-Dong; Li, Hong; Han, Yan

    2016-12-30

    BACKGROUND CC chemokine receptor 7 (CCR7) expression is vital for cell migration to secondary lymphoid organs (SLOs). Our previous work showed that inducing CCR7 expression enabled syngeneic mesenchymal stem cells (MSCs) to migrate into SLOs, resulting in enhanced immunosuppressive performance in mice. Given that human adipose-derived stem cells (hASCs) are widely used in clinical therapy, we further investigated whether upregulation of CCR7 enables xenogeneic hASCs to migrate to rat SLOs. MATERIAL AND METHODS hASCs rarely express CCR7; therefore, hASCs were transfected with lentivirus encoding rat CCR7 (rCCR7) plus green fluorescence protein (GFP) or GFP alone. CCR7 mRNA and cell surface expression of rCCR7-hASCs and GFP-hASCs were examined by reverse transcription-polymerase chain reaction (RT-PCR) and flow cytometry (FCM), respectively. The phenotype, differentiation, and proliferation capacity of each cell type was also determined. To examine migration, rCCR7-hASCs and GFP-hASCs were injected intravenously into Lewis rats, and the proportion of GFP-positive cells in the spleen and lymph nodes was determined with FCM. RESULTS mRNA and cell surface protein expression of CCR7 was essentially undetectable in hASCs and GFP-ASCs; however, CCR7 was highly expressed in rCCR7-ASCs. rCCR7-hASCs, GFP-hASCs, and hASCs shared a similar immunophenotype, and maintained the ability of multilineage differentiation and proliferation. In addition, the average proportion of GFP-positive cells was significantly higher following transplantation of rCCR7-hASCs compared with GFP-hASCs (p<0.01). CONCLUSIONS These results suggest that upregulation of rat CCR7 expression does not change the phenotype, differentiation, or proliferation capacity of hASCs, but does enable efficient migration of hASCs to rat SLOs.

  6. Comparison of drug and cell-based delivery: engineered adult mesenchymal stem cells expressing soluble tumor necrosis factor receptor II prevent arthritis in mouse and rat animal models.

    Science.gov (United States)

    Liu, Linda N; Wang, Gang; Hendricks, Kyle; Lee, Keunmyoung; Bohnlein, Ernst; Junker, Uwe; Mosca, Joseph D

    2013-05-01

    Rheumatoid arthritis (RA) is a systemic autoimmune disease with unknown etiology where tumor necrosis factor-α (TNFα) plays a critical role. Etanercept, a recombinant fusion protein of human soluble tumor necrosis factor receptor II (hsTNFR) linked to the Fc portion of human IgG1, is used to treat RA based on the rationale that sTNFR binds TNFα and blocks TNFα-mediated inflammation. We compared hsTNFR protein delivery from genetically engineered human mesenchymal stem cells (hMSCs) with etanercept. Blocking TNFα-dependent intercellular adhesion molecule-1 expression on transduced hMSCs and inhibition of nitric oxide production from TNFα-treated bovine chondrocytes by conditioned culture media from transduced hMSCs demonstrated the functionality of the hsTNFR construction. Implanted hsTNFR-transduced mesenchymal stem cells (MSCs) reduced mouse serum circulating TNFα generated from either implanted TNFα-expressing cells or lipopolysaccharide induction more effectively than etanercept (TNFα, 100%; interleukin [IL]-1α, 90%; and IL-6, 60% within 6 hours), suggesting faster clearance of the soluble tumor necrosis factor receptor (sTNFR)-TNFα complex from the animals. In vivo efficacy of sTNFR-transduced MSCs was illustrated in two (immune-deficient and immune-competent) arthritic rodent models. In the antibody-induced arthritis BalbC/SCID mouse model, intramuscular injection of hsTNFR-transduced hMSCs reduced joint inflammation by 90% compared with untransduced hMSCs; in the collagen-induced arthritis Fischer rat model, both sTNFR-transduced rat MSCs and etanercept inhibited joint inflammation by 30%. In vitro chondrogenesis assays showed the ability of TNFα and IL1α, but not interferon γ, to inhibit hMSC differentiation to chondrocytes, illustrating an additional negative role for inflammatory cytokines in joint repair. The data support the utility of hMSCs as therapeutic gene delivery vehicles and their potential to be used in alleviating inflammation

  7. Magnetic resonance imaging tracing of transplanted bone marrow mesenchymal stem cells in a rat model of cardiac arrest-induced global brain ischemia

    Institute of Scientific and Technical Information of China (English)

    Yue Fu; Xiangshao Fang; Tong Wang; Jiwen Wang; Jun Jiang; Zhigang Luo; Xiaohui Duan; Jun Shen; Zitong Huang

    2009-01-01

    BACKGROUND: Numerous studies have shown that magnetic resonance imaging (MRI) can detect survival and migration of super paramagnetic iron oxide-labeled stem cells in models of focal cerebral infarction. OBJECTIVE: To observe distribution of bone marrow mesenchymal stem cells (BMSCs) in a rat model of global brain ischemia following cardiac arrest and resuscitation, and to investigate the feasibility of tracing iron oxide-labeled BMSCs using non-invasive MRI. DESIGN, TIME AND SETTING: The randomized, controlled, molecular imaging study was performed at the Linbaixin Medical Research Center, Second Affiliated Hospital, Sun Yat-sen University, and the Institute of Cardiopulmonary Cerebral Resuscitation, Sun Yat-sen University, China from October 2006 to February 2009.MATERIALS: A total of 40 clean, Sprague Dawley rats, aged 6 weeks and of either gender, were supplied by the Experimental Animal Center, Sun Yat-sen University, China, for isolation of BMSCs. Feridex (iron oxide), Gyroscan Inetra 1.5T MRI system, and cardiopulmonary resuscitation device were used in this study. METHODS: A total of 30 healthy, male Sprague Dawley rats, aged 6 months, were used to induce ventricular fibrillation using alternating current. After 8 minutes, the rats underwent 6-minute chest compression and mechanical ventilation, followed by electric defibrillation, to establish rat models of global brain ischemia due to cardiac arrest and resuscitation. A total of 24 successful models were randomly assigned to Feridex-labeled and non-labeled groups (n=12 for each group). At 2 hours after resuscitation, 5 x 10 6 Feddex-labeled BMSCs, with protamine sulfate as a carrier, and 5 × 10 6 non-labeled BMSCs were respectively transplanted into both groups of rats through the right carotid artery (cells were harvested in 1 mL phosphate buffered saline). MAIN OUTCOME MEASURES: Feridex-labeled BMSCs were observed by Prussian blue staining and electron microscopy. Signal intensity, celluar viability

  8. In vitro cultivation of rat bone marrow mesenchymal stem cells and establishment of pEGFP/Ang-1 transfection method

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    Xiu-Qun Zhang

    2014-09-01

    Conclusions: Adherence screening method and density gradient centrifugation can be effective methods to obtain BMSCs with high purity and rapid proliferation. Besides, the expression of transfected recombinant plasmid pEGFP/Ang-1 in rat BMSCs is satisfactory.

  9. Effect of fatty acid methyl esters from plastrum testudinis on proliferation of rat bone mesenchymal stem cells

    Institute of Scientific and Technical Information of China (English)

    Yuehua ZHANG; Heping ZENG; Dongfeng CHEN

    2008-01-01

    The ointment of plastrum testudinis was extracted using petroleum ether,ether and dichloromethane sequentially and the extracts were methyl,esteri,fled. The effects on the proliferation of bone marrow mesenchymal stem cells (bMMSCs) were examined by MTT[3,(4,5,dimethylthiazol,2,yl),2,5,diphenyl tetrazolium bromide] assay and flow cytometry analysis. The volatile components of the samples were studied by gas chromatography,mass spectrometry (GC,MS) and high performance liquid chromatography (HPLC). The results show that the methyl,esterified parts can promote the proliferation of stem cells and they all contain palmitic acid methyl ester. Palmitic acid methyl ester can promote proliferation when the concentration was 0.15 μg/μL. It may be concluded that the palmitic acid methyl ester is important for the methyl,esterified parts that have effects on proliferation.

  10. CXCR-7 receptor promotes SDF-1α-induced migration of bone marrow mesenchymal stem cells in the transient cerebral ischemia/reperfusion rat hippocampus.

    Science.gov (United States)

    Wang, Yulan; Fu, Wei; Zhang, Shichun; He, Xiaomei; Liu, Zhi'an; Gao, Diansuai; Xu, Tiejun

    2014-08-05

    The stromal cell-derived factor 1/C-X-C chemokine receptor type 4 (SDF-1/CXCR-4) axis plays an important role during stem cell recruitment. SDF-1 can also bind the more recently described CXCR-7 receptor, but effects of SDF-1/CXCR-7 signaling on stem cell migrating to ischemic brain injury area are little known. In the present study, we investigated the effect of CXCR-7 on bone marrow mesenchymal stem cell (BMSC) migration toward SDF-1α in the cerebral ischemia/reperfusion (I/R) rat hippocampus. We cultured BMSCs from rats and characterized them using flow cytometry, immunocytochemistry, western blotting, and immunofluorescence to detect SDF-1α, CXCR-4, and CXCR-7 expression in third passage BMSCs (P3-BMSCs). We also prepared the model of transient cerebral I/R by four-vessel occlusion (4-VO), and BMSCs were transplanted into I/R rat brain via lateral ventricle (LV) injection (20μl, 1×10(6)/ml). After that, we examined the effect of BMSCs migration in the cerebral I/R rat hippocampus through Transwell chamber assay. Our results show that SDF-1α, CXCR-4, and CXCR-7 were expressed in P3-BMSCs. Moreover, SDF-1α expression was increased in I/R hippocampus. At 48h after transplant, green fluorescent BrdU-BMSCs were observed in transplant groups, but no green fluorescent BrdU-BMSCs were seen in medium group. Among BMSCs transplant groups, the number of BrdU-BMSCs positive cell was the highest in BMSC group. Treatment with AMD3100 and/or CXCR-7 neutralizing antibody decreased the number of BMSC migration. Collectively, these findings indicate that CXCR-4 and -7 receptors were co-expressed in BMSCs and synergistically promoted BMSC migration. The effect of CXCR-7 was stronger than that of CXCR-4. Moreover, BMSCs that migrated to hippocampus promoted the autocrine and paracrine signaling of SDF-1α.

  11. Acute and subacute chemical-induced lung injuries: HRCT findings

    Energy Technology Data Exchange (ETDEWEB)

    Akira, Masanori, E-mail: Akira@kch.hosp.go.jp [Department of Radiology, National Hospital Organization Kinki-Chuo Chest Medical Center, 1180 Nagasone-cho, Kita-ku, Sakai City, Osaka 591-8555 (Japan); Suganuma, Narufumi [Department of Environmental Medicine, Kochi Medical School (Japan)

    2014-08-15

    Lung injury caused by chemicals includes bronchitis, bronchiolitis, chemical pneumonitis, pulmonary edema, acute respiratory distress syndrome, organizing pneumonia, hypersensitivity pneumonitis, acute eosinophilic pneumonia, and sarcoid-like granulomatous lung disease. Each chemical induces variable pathophysiology and the situation resembles to the drug induced lung disease. The HRCT features are variable and nonspecific, however HRCT may be useful in the evaluation of the lung injuries and so we should know about HRCT features of lung parenchymal abnormalities caused by chemicals.

  12. Chemically induced electric field: flat band potential engineering

    Science.gov (United States)

    Bak, T.; Guo, Z.; Li, W.; Atanacio, A. J.; Nowotny, J.

    2012-10-01

    The present work considers engineering of the flat band potential, FBP, of metal oxides in a controlled manner. The aim is to minimise the energy losses related to recombination. The related experimental approaches include imposition of a chemically-induced electric field using the phenomena of segregation, diffusion and the formation of multilayer systems. This paper considers several basic phenomena that allow the modification of the surface charge and the space charge at the gas/solid and solid/liquid interfaces.

  13. Low-level laser therapy with helium-neon laser improved viability of osteoporotic bone marrow-derived mesenchymal stem cells from ovariectomy-induced osteoporotic rats

    Science.gov (United States)

    Fallahnezhad, Somaye; Piryaei, Abbas; Tabeie, Faraj; Nazarian, Hamid; Darbandi, Hasan; Amini, Abdoldllah; Mostafavinia, Ataroalsadat; Ghorishi, Seyed Kamran; Jalalifirouzkouhi, Ali; Bayat, Mohammad

    2016-09-01

    The purpose of this study was to evaluate the influences of helium-neon (He-Ne) and infrared (IR) lasers on the viability and proliferation rate of healthy and ovariectomy-induced osteoporotic (OVX) bone marrow mesenchymal stem cells (BMMSCs) in vitro. MSCs harvested from the BM of healthy and OVX rats were culture expanded. He-Ne and IR lasers were applied three times at energy densities of 0.6, 1.2, and 2.4 J/cm2 for BMMSCs. BMMSCs viability and proliferation rate were evaluated by MTT assay on days 2, 4, 6, 14, and 21. The results showed that healthy BMMSCs responded optimally to 0.6 J/cm2 using an IR laser after three times of laser radiation. Moreover, it was found that OVX-BMMSCs responded optimally to 0.6 J/cm2 with He-Ne laser and one-time laser radiation. It is concluded that the low-level laser therapy (LLLT) effect depends on the physiological state of the BMMSCs, type of the laser, wavelength, and number of laser sessions. The biostimulation efficiency of LLLT also depends on the delivered energy density. LLLT can enhance the viability and proliferation rate of healthy and especially osteoporotic autologous BMMSCs, which could be very useful in regenerative medicine.

  14. Recruitment of exogenous mesenchymal stem cells in mandibular distraction osteogenesis by the stromal cell-derived factor-1/chemokine receptor-4 pathway in rats.

    Science.gov (United States)

    Cao, Jian; Wang, Lei; Du, Zhao-jie; Liu, Peng; Zhang, Ya-bo; Sui, Jian-fu; Liu, Yan-pu; Lei, De-lin

    2013-12-01

    Distraction osteogenesis is widely used in orthopaedic and craniofacial surgery. However, its exact mechanism is still poorly understood. The purpose of this study was to find out whether there is systemic recruitment of mesenchymal stem cells (MSC) to the neocallus in the distraction gap by the stromal cell-derived factor-1 (SDF-1)/CXC chemokine receptor 4 (CXCR4) axis during osteogenesis. We examined the migration of MSC towards a gradient of SDF-1 in vitro. We also transplanted MSC labelled with green fluorescent protein (GFP) intravenously, with or without treatment with CXCR4-blocking antibody, into rats that had had unilateral mandibular distraction osteogenesis, and investigated the distribution of cells labelled with GFP in the soft callus after 24 h. We found that SDF-1 facilitated the migration potency of MSC both in vitro and in vivo, and this migration could be inhibited by AMD3100, an antagonist of CXCR4, and promoted by local infusion of exogenous SDF-1 into the distraction gap. This study provides a new insight into the molecular basis of how new bone is regenerated during distraction osteogenesis.

  15. MiR-124 suppresses the chemotactic migration of rat mesenchymal stem cells toward HGF by downregulating Wnt/β-catenin signaling.

    Science.gov (United States)

    Yue, Qing; Zhang, Yu; Li, Xianyang; He, Lihong; Hu, Ya'nan; Wang, Xianyao; Xu, Xiaojing; Shen, Yixin; Zhang, Huanxiang

    2016-09-01

    Mesenchymal stem cells (MSCs) exhibit the potential to repair a wide variety of injured adult tissues. The migration capability of MSCs is an important determinant of the efficiency of MSC transplant therapy. MicroRNAs (miRNAs) are increasingly implicated in regulating the migration of MSCs. Herein, we show that the expression of miR-124 was downregulated in rat MSCs (rMSCs) treated with hepatocyte growth factor (HGF). Overexpression of miR-124 significantly reduced the chemotactic migration of rMSCs toward HGF, while inhibition of endogenous miR-124 promoted the chemotactic migration. A further study revealed that miR-124 directly targeted FZD4 and LRP6, which encode a receptor and co-receptor of the Wnt/β-catenin signaling pathway, respectively, thus reducing the activity of this signaling. Consistently, activation of the Wnt/β-catenin signaling pathway by LiCl and ΔN89β-catenin rescued the inhibitory effect of miR-124 on the chemotactic migration of rMSCs toward HGF, while inhibition of Wnt/β-catenin signaling by FH535 abrogated the enhanced chemotactic response achieved by the miR-124 inhibitor. Collectively, our study demonstrates that miR-124 downregulates Wnt/β-catenin signaling via targeting FZD4 and LRP6 and thus suppresses the chemotactic migration of rMSCs toward HGF.

  16. Combined Effects of Mechanical Strain and Hydroxyapatite/Collagen Composite on Osteogenic Differentiation of Rat Bone Marrow Derived Mesenchymal Stem Cells

    Directory of Open Access Journals (Sweden)

    Yan Huang

    2013-01-01

    Full Text Available Mesenchymal stem cells (MSCs represent a promising source for bone repair and regeneration. Recent lines of evidence have shown that appropriate strain could regulate the osteogenic differentiation of MSCs. Our previous study demonstrated that hydroxyapatite/collagen (HA/Col composite also played an important role in the osteogenic differentiation of MSCs. The aim of this study is to investigate the effects of mechanical strain and HA/Col composite on the osteogenic differentiation of rat bone marrow derived MSCs (rBMSCs in vitro. rBMSCs were treated with cyclic strain generated by a self-designed stretching device with or without the presence of HA/Col composite. Osteogenic differentiation levels were evaluated using reverse transcription polymerase chain reaction (RT-PCR, alkaline phosphatase spectrophotometry, and western blotting. The results demonstrated that mechanical strain combined with HA/Col composite could obviously induce the differentiation of rBMSCs into osteoblasts, which had a better effect than only mechanical strain or HA/Col composite treatment. This provides a new avenue for mechanistic studies of stem cell differentiation and a novel approach to obtain more committed differentiated cells.

  17. Delivery of human mesenchymal adipose-derived stem cells restores multiple urological dysfunctions in a rat model mimicking radical prostatectomy damages through tissue-specific paracrine mechanisms.

    Science.gov (United States)

    Yiou, René; Mahrouf-Yorgov, Meriem; Trébeau, Céline; Zanaty, Marc; Lecointe, Cécile; Souktani, Richard; Zadigue, Patricia; Figeac, Florence; Rodriguez, Anne-Marie

    2016-02-01

    Urinary incontinence (UI) and erectile dysfunction (ED) are the most common functional urological disorders and the main sequels of radical prostatectomy (RP) for prostate cancer. Mesenchymal stem cell (MSC) therapy holds promise for repairing tissue damage due to RP. Because animal studies accurately replicating post-RP clinical UI and ED are lacking, little is known about the mechanisms underlying the urological benefits of MSC in this setting. To determine whether and by which mechanisms MSC can repair damages to both striated urethral sphincter (SUS) and penis in the same animal, we delivered human multipotent adipose stem cells, used as MSC model, in an immunocompetent rat model replicating post-RP UI and ED. In this model, we demonstrated by using noninvasive methods in the same animal from day 7 to day 90 post-RP injury that MSC administration into both the SUS and the penis significantly improved urinary continence and erectile function. The regenerative effects of MSC therapy were not due to transdifferentiation and robust engraftment at injection sites. Rather, our results suggest that MSC benefits in both target organs may involve a paracrine process with not only soluble factor release by the MSC but also activation of the recipient's secretome. These two effects of MSC varied across target tissues and damaged-cell types. In conclusion, our work provides new insights into the regenerative properties of MSC and supports the ability of MSC from a single source to repair multiple types of damage, such as those seen after RP, in the same individual.

  18. Influence of the intensity and loading time of direct current electric field on the directional migration of rat bone marrow mesenchymal stem cells.

    Science.gov (United States)

    Wang, Xiaoyu; Gao, Yuxuan; Shi, Haigang; Liu, Na; Zhang, Wei; Li, Hongbo

    2016-09-01

    Exogenic electric fields can effectively accelerate bone healing and remodeling through the enhanced migration of bone marrow mesenchymal stem cells (BMSCs) toward the injured area. This study aimed to determine the following: (1) the direction of rat BMSC (rBMSC) migration upon exposure to a direct current electric field (DCEF), (2) the optimal DCEF intensity and duration, and (3) the possible regulatory role of SDF-1/CXCR4 axis in rBMSC migration as induced by DCEF. Results showed that rBMSCs migrated to the positive electrode of the DCEF, and that the DCEF of 200 mV/mm for 4 h was found to be optimal in enhancing rBMSC migration. This DCEF strength and duration also upregulated the expression of osteoblastic genes, including ALP and OCN, and upregulated the expression of ALP and Runx2 proteins. Moreover, when CXCR4 was inhibited, rBMSC migration due to DCEF was partially blocked. These findings indicated that DCEF can effectively induce rBMSC migration. A DCEF of 200 mV/mm for 4 h was recommended because of its ability to promote rBMSC migration, proliferation, and osteogenic differentiation. The SDF-1/CXCR4 signaling pathway may play an important role in regulating the DCEF-induced migration of rBMSCs.

  19. Mesenchymal Stem Cell Attenuates Neutrophil-predominant Inflammation and Acute Lung Injury in an In Vivo Rat Model of Ventilator-induced Lung Injury

    Directory of Open Access Journals (Sweden)

    Tian-Shun Lai

    2015-01-01

    Full Text Available Background: Subsequent neutrophil (polymorphonuclear neutrophil [PMN]-predominant inflammatory response is a predominant feature of ventilator-induced lung injury (VILI, and mesenchymal stem cell (MSC can improve mice survival model of endotoxin-induced acute lung injury, reduce lung impairs, and enhance the repair of VILI. However, whether MSC could attenuate PMN-predominant inflammatory in the VILI is still unknown. This study aimed to test whether MSC intervention could attenuate the PMN-predominate inflammatory in the mechanical VILI. Methods: Sprague-Dawley rats were ventilated for 2 hours with large tidal volume (20 mL/kg. MSCs were given before or after ventilation. The inflammatory chemokines and gas exchange were observed and compared dynamically until 4 hours after ventilation, and pulmonary pathological change and activation of PMN were observed and compared 4 hours after ventilation. Results: Mechanical ventilation (MV caused significant lung injury reflected by increasing in PMN pulmonary sequestration, inflammatory chemokines (tumor necrosis factor-alpha, interleukin-6 and macrophage inflammatory protein 2 in the bronchoalveolar lavage fluid, and injury score of the lung tissue. These changes were accompanied with excessive PMN activation which reflected by increases in PMN elastase activity, production of radical oxygen series. MSC intervention especially pretreatment attenuated subsequent lung injury, systemic inflammation response and PMN pulmonary sequestration and excessive PMN activation initiated by injurious ventilation. Conclusions: MV causes profound lung injury and PMN-predominate inflammatory responses. The protection effect of MSC in the VILI rat model is related to the suppression of the PMN activation.

  20. Reconstruction of rat calvarial defects with human mesenchymal stem cells and osteoblast-like cells in poly-lactic-co-glycolic acid scaffolds

    Directory of Open Access Journals (Sweden)

    C Zong

    2010-09-01

    Full Text Available Human mesenchymal stem cells (hMSCs can be used for xenogenic transplantation due to their low immunogenicity, high proliferation rate, and multi-differentiation potentials. Therefore, hMSCs are an ideal seeding source for tissue engineering. The present study evaluates the reconstruction effects of hMSCs and osteoblast-like cells differentiated from hMSCs in poly-lactic-co-glycolic acid (PLGA scaffolds on the calvarial defect of rats. Two bilateral full-thickness defects (5mm in diameter were created in the calvarium of nonimmunosuppressed Sprague-Dawley rats. The defects were filled by PLGA scaffolds with hMSCs (hMSC Construct or with osteoblast-like cells differentiated from hMSCs (Osteoblast Construct. The defects without any graft (Blank Defect or filled with PLGA scaffold without any cells (Blank Scaffold were used as controls. Evaluation was performed using macroscopic view, histology and immunohistochemical analysis respectively at 10 and 20 weeks after transplantation. In addition, fluorescent carbocyanine CM-Dil was used to track the implanted cells in vivo during transplantation. The results showed that while both hMSC Construct and Osteoblast Construct led to an effective reconstruction of critical-size calvarial defects, the bone reconstruction potential of hMSC Construct was superior to that of Osteoblast Construct in non-autogenous applications. Our findings verify the feasibility of the use of xenogenic MSCs for tissue engineering and demonstrate that undifferentiated hMSCs are more suitable for bone reconstruction in xenotransplantation models.

  1. Mesenchymal Stem Cell Attenuates Neutrophil-predominant Inflammation and Acute Lung Injury in an In Vivo Rat Model of Ventilator-induced Lung Injury

    Institute of Scientific and Technical Information of China (English)

    Tian-Shun Lai; Zhi-Hong Wang; Shao-Xi Cai

    2015-01-01

    Background:Subsequent neutrophil (polymorphonuclear neutrophil [PMN])-predominant inflammatory response is a predominant feature of ventilator-induced lung injury (VILI),and mesenchymal stem cell (MSC) can improve mice survival model of endotoxin-induced acute lung injury,reduce lung impairs,and enhance the repair of VILI.However,whether MSC could attenuate PMN-predominant inflammatory in the VILI is still unknown.This study aimed to test whether MSC intervention could attenuate the PMN-predominate inflammatory in the mechanical VILI.Methods:Sprague-Dawley rats were ventilated for 2 hours with large tidal volume (20 mL/kg).MSCs were given before or after ventilation.The inflammatory chemokines and gas exchange were observed and compared dynamically until 4 hours after ventilation,and pulmonary pathological change and activation of PMN were observed and compared 4 hours after ventilation.Results:Mechanical ventilation (MV) caused significant lung injury reflected by increasing in PMN pulmonary sequestration,inflammatory chemokines (tumor necrosis factor-alpha,interleukin-6 and macrophage inflammatory protein 2) in the bronchoalveolar lavage fluid,and injury score of the lung tissue.These changes were accompanied with excessive PMN activation which reflected by increases in PMN elastase activity,production of radical oxygen series.MSC intervention especially pretreatment attenuated subsequent lung injury,systemic inflammation response and PMN pulmonary sequestration and excessive PMN activation initiated by injurious ventilation.Conclusions:MV causes profound lung injury and PMN-predominate inflammatory responses.The protection effect of MSC in the VILI rat model is related to the suppression of the PMN activation.

  2. Nanoparticle Labeling of Bone Marrow-Derived Rat Mesenchymal Stem Cells: Their Use in Differentiation and Tracking

    Directory of Open Access Journals (Sweden)

    Ece Akhan

    2015-01-01

    Full Text Available Mesenchymal stem cells (MSCs are promising candidates for cellular therapies due to their ability to migrate to damaged tissue without inducing immune reaction. Many techniques have been developed to trace MSCs and their differentiation efficacy; however, all of these methods have limitations. Conjugated polymer based water-dispersible nanoparticles (CPN represent a new class of probes because they offer high brightness, improved photostability, high fluorescent quantum yield, and noncytotoxicity comparing to conventional dyes and quantum dots. We aimed to use this tool for tracing MSCs’ fate in vitro and in vivo. MSC marker expression, survival, and differentiation capacity were assessed upon CPN treatment. Our results showed that after CPN labeling, MSC markers did not change and significant number of cells were found to be viable as revealed by MTT. Fluorescent signals were retained for 3 weeks after they were differentiated into osteocytes, adipocytes, and chondrocytes in vitro. We also showed that the labeled MSCs migrated to the site of injury and retained their labels in an in vivo liver regeneration model. The utilization of nanoparticle could be a promising tool for the tracking of MSCs in vivo and in vitro and therefore can be a useful tool to understand differentiation and homing mechanisms of MSCs.

  3. Safrole oxide induced neuronal differentiation of rat bone-marrow mesenchymal stem cells by elevating Hsp70.

    Science.gov (United States)

    Zhao, YanChun; Xin, Jie; Sun, ChunHui; Zhao, BaoXiang; Zhao, Jing; Su, Le

    2012-11-01

    In a previous study, we found that at low concentrations, safrole oxide (SFO) could induce vascular endothelial cell (VEC) transdifferentiation into neuron-like cells; however, whether SFO could induce bone-marrow mesenchymal stem cell (BMSC) neural differentiation was unknown. Here, we found that SFO could effectively induce BMSC neural differentiation in the presence of serum and fibroblast growth factor 2 and did not affect cell viability at low concentrations. The levels of neuron-specific enolase and neurofilament-L were increased greatly, but that of glial fibrillary acidic protein was absent with SFO treatment for 48h. Furthermore, SFO could increase the level of heat shock protein 70 (Hsp70), an important factor in neuronal differentiation. Knockdown of Hsp70 by its small interfering RNA blocked SFO-induced BMSC differentiation. Thus, SFO is a novel inducer of BMSC differentiation to neuron-like cells and Hsp70 is implicated in the differentiation process. We provide a new tool for obtaining neuron-like cells from BMSCs and for further investigating the new effect of Hsp70 on BMSC neuronal differentiation.

  4. TiO2-Nanowired Delivery of Mesenchymal Stem Cells Thwarts Diabetes- Induced Exacerbation of Brain Pathology in Heat Stroke: An Experimental Study in the Rat Using Morphological and Biochemical Approaches.

    Science.gov (United States)

    Sharma, Hari S; Feng, Lianyuan; Lafuente, José V; Muresanu, Dafin F; Tian, Zhenrong R; Patnaik, Ranjana; Sharma, Aruna

    2015-01-01

    We have shown previously that heat stroke produced by whole body hyperthermia (WBH) for 4 h at 38°C in diabetic rats exacerbates blood-brain barrier breakdown, brain edema formation and neuronal cell injury as compared to healthy animals after identical heat exposure. In this combination of diabetes and WBH, normal therapeutic measures do not induce sufficient neuroprotection. Thus, we investigated whether nanowired mesenchymal cells (MSCs) when delivered systemically may have better therapeutic effects on brain damage in diabetic rats after WBH. Diabetes induced by streptozotocin administration (75 mg/kg, i.p, daily for 3 days) in rats resulted in clinical symptoms of the disease within 4 to 6 weeks (blood glucose level 20 to 30 mmoles/l as compared to saline control groups (4 to 6 mmoles/l). When subjected to WBH, these diabetic rats showed a 4-to 6-fold exacerbation of blood-brain barrier breakdown to Evans blue and radioiodine, along with brain edema formation and neuronal cell injury. Intravenous administration of rat MSCs (1x10(6)) to diabetic rats one week before WBH slightly reduced brain pathology, whereas TiO2 nanowired MSCs administered in an identical manner resulted in almost complete neuroprotection. On the other hand, MSCs alone significantly reduced brain pathology in saline-treated rats after WBH. These observations indicate that nanowired delivery of stem cells has superior therapeutic potential in heat stroke with diabetes, pointing to novel clinical perspectives in the future.

  5. Pre-treatment with bone marrow-derived mesenchymal stem cells inhibits systemic intravascular coagulation and attenuates organ dysfunction in lipopolysaccharide-induced disseminated intravascular coagulation rat model

    Institute of Scientific and Technical Information of China (English)

    WANG Biao; WU Shu-ming; WANG Tao; LIU Kai; ZHANG Gong; ZHANG Xi-quan; YU Jian-hua; LIU Chuan-zhen; FANG Chang-cun

    2012-01-01

    Background Bacterial lipopolysaccharide (LPS) can activate immunological cells to secrete various proinflammatory cytokines involved in the pathophysiological process of disseminated intravascular coagulation (DIC) during infection.In recent years,it has been found that bone marrow-derived mesenchymal stem cells (BMSCs) can affect the activity of these immune cells and regulate the secretion of proinflammatory cytokines.Here,we report the possible protective effect of BMSCs pre-treatment in LPS-induced DIC rat model and the mechanism.Methods Forty-eight adult male rats were divided into five experimental groups and one control group with eight animals in each group.In the treatment groups,0,1×106,2×106,3×106,and 5×106 of BMSCs were injected intravenously for 3 days before LPS injection,while the control group was treated with pure cell culture medium injection.Then,the LPS (3 mg/kg) was injected via the tail vein in the treatment groups,while the control group received 0.9% NaCl.Blood was withdrawn before and 4 and 8 hours after LPS administration.The following parameters were monitored:platelets (PLT),fibrinogen (Fib),D-dimer (D-D),activated partial thromboplastin time (APTT),prothrombin time (PT),tumor necrosis factor-o (TNF-α),interferon-γ (IFN-γ),interleukin-1β (IL-1β),creatinine (Cr),alanine aminotransferase (ALT),creatinine kinase-MB (CK-MB),and endothelin (ET).Results Compared with the control group,a significant change of coagulation parameters were found in the experimental groups.The plasma level of the inflammatory mediator (TNF-α,IFN-γ,IL-1β),organ indicator (Cr,ALT,and CK-MB),and ET in the experimental groups were much lower (P <0.05) than that in the control group.Furthermore,some of these effects were dose-dependent; the statistical comparison of the plasma levels between the groups (from group 2 to group 5) showed a significant difference (P<0.05),except the ALT and CK-MB levels (P>0.05).Conclusion Pre-treatment with BMSCs can

  6. In-vivo generation of bone via endochondral ossification by in-vitro chondrogenic priming of adult human and rat mesenchymal stem cells

    LENUS (Irish Health Repository)

    Farrell, Eric

    2011-01-31

    Abstract Background Bone grafts are required to repair large bone defects after tumour resection or large trauma. The availability of patients\\' own bone tissue that can be used for these procedures is limited. Thus far bone tissue engineering has not lead to an implant which could be used as alternative in bone replacement surgery. This is mainly due to problems of vascularisation of the implanted tissues leading to core necrosis and implant failure. Recently it was discovered that embryonic stem cells can form bone via the endochondral pathway, thereby turning in-vitro created cartilage into bone in-vivo. In this study we investigated the potential of human adult mesenchymal stem cells to form bone via the endochondral pathway. Methods MSCs were cultured for 28 days in chondrogenic, osteogenic or control medium prior to implantation. To further optimise this process we induced mineralisation in the chondrogenic constructs before implantation by changing to osteogenic medium during the last 7 days of culture. Results After 8 weeks of subcutaneous implantation in mice, bone and bone marrow formation was observed in 8 of 9 constructs cultured in chondrogenic medium. No bone was observed in any samples cultured in osteogenic medium. Switch to osteogenic medium for 7 days prevented formation of bone in-vivo. Addition of β-glycerophosphate to chondrogenic medium during the last 7 days in culture induced mineralisation of the matrix and still enabled formation of bone and marrow in both human and rat MSC cultures. To determine whether bone was formed by the host or by the implanted tissue we used an immunocompetent transgenic rat model. Thereby we found that osteoblasts in the bone were almost entirely of host origin but the osteocytes are of both host and donor origin. Conclusions The preliminary data presented in this manuscript demonstrates that chondrogenic priming of MSCs leads to bone formation in vivo using both human and rat cells. Furthermore, addition of

  7. Mesenchymal progenitor cells differentiate into an endothelial phenotype, enhance vascular density and improve heart function in a rat cellular cardiomyoplasty model

    Institute of Scientific and Technical Information of China (English)

    SDAVANI; NMERSIN; BROYER; BKANTELIP; JPKANTELIP

    2004-01-01

    AIM: Cellular cardiomyoplasty is promising for improving postinfarcted cardiac function. Over the past decade, a variety of cell types have been proposed including mononuclear bone marrow cells. The latter contains different lineages including mesenchymal stem cells (MSCs). The aim of this study was to analyse the differentiation pathways of engrafted syngenic mesenchymal progenitor cells (MPCs) obtained in culture from bone marrow

  8. Bone mesenchymal stem cell transplantation via four routes for the treatment of acute liver failure in rats.

    Science.gov (United States)

    Sun, Lihua; Fan, Xiaotang; Zhang, Lijuan; Shi, Guixiu; Aili, Maimaiti; Lu, Xiaobo; Jiang, Tao; Zhang, Yuexin

    2014-10-01

    In the present study, we assessed the efficiency of four BMSC transplantation methods as a therapy for liver failure. A rat model (80 Sprague-Dawley rats) of D-galactosamine (D-gal)/lipopolysaccharide (LPS)-induced acute liver failure (ALF) was established and the rats were divided into 5 groups: a hepatic artery injection group, a portal vein injection group, a vena caudalis injection group, an intraperitoneal injection group and a control group (16 per group). Following transplantation, the liver tissue and blood samples were collected on days 1, 3 and 7, we detected the EdU (5-ethynyl-2'-deoxyuridine)-labeled cells homing to the liver tissue and assessed the proliferating cell nuclear antigen (PCNA) and cysteine-containing aspartate-specific protease (caspase)-3 expression in the liver tissue and detected the levels of stromal cell-derived factor 1 (SDF-1) and hepatocyte growth factor (HGF) in the liver tissues. Compared with the control group, the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and damage to the liver tissue in the hepatic artery group, the portal vein group and the vena caudalis group improved in vivo. The expression of PCNA and HGF in the liver was higher and caspase-3 expression was lower in the hepatic artery injection group, the portal vein injection group and the vena caudalis injection group than that in the intraperitoneal injection and control groups. The EdU-labeled BMSCs were only observed homing to the liver tissue in these three groups. However, no significant differences were observed between these three groups. Liver function in the rats with ALF was improved following BMSC transplantation via 3 endovascular implantation methods (through the hepatic artery, portal vein and vena caudalis). These 3 methods were effective in transplanting BMSCs for the treatment of ALF. However, the selection of blood vessel in the implantation pathway does not affect the transplantation outcome. Transplantation via

  9. Chemically induced intestinal damage models in zebrafish larvae.

    Science.gov (United States)

    Oehlers, Stefan H; Flores, Maria Vega; Hall, Christopher J; Okuda, Kazuhide S; Sison, John Oliver; Crosier, Kathryn E; Crosier, Philip S

    2013-06-01

    Several intestinal damage models have been developed using zebrafish, with the aim of recapitulating aspects of human inflammatory bowel disease (IBD). These experimentally induced inflammation models have utilized immersion exposure to an array of colitogenic agents (including live bacteria, bacterial products, and chemicals) to induce varying severity of inflammation. This technical report describes methods used to generate two chemically induced intestinal damage models using either dextran sodium sulfate (DSS) or trinitrobenzene sulfonic acid (TNBS). Methods to monitor intestinal damage and inflammatory processes, and chemical-genetic methods to manipulate the host response to injury are also described.

  10. Chemically induced magnetism in atomically precise gold clusters.

    Science.gov (United States)

    Krishna, Katla Sai; Tarakeshwar, Pilarisetty; Mujica, Vladimiro; Kumar, Challa S S R

    2014-03-12

    Comparative theoretical and experimental investigations are reported into chemically induced magnetism in atomically-precise, ligand-stabilized gold clusters Au25 , Au38 and Au55 . The results indicate that [Au25 (PPh3 )10 (SC12 H25 )5 Cl2 ](2+) and Au38 (SC12 H25 )24 are diamagnetic, Au25 (SC2 H4 Ph)18 is paramagnetic, and Au55 (PPh3 )12 Cl6 , is ferromagnetic at room temperature. Understanding the magnetic properties resulting from quantum size effects in such atomically precise gold clusters could lead to new fundamental discoveries and applications.

  11. Boron Induces Early Matrix Mineralization via Calcium Deposition and Elevation of Alkaline Phosphatase Activity in Differentiated Rat Bone Marrow Mesenchymal Stem Cells

    Directory of Open Access Journals (Sweden)

    Bent-al-hoda Movahedi Najafabadi

    2016-04-01

    Full Text Available Objective: Boron (B is essential for plant development and might be an essential micronutrient for animals and humans. This study was conducted to characterize the impact of boric acid (BA on the cellular and molecular nature of differentiated rat bone marrow mesenchymal stem cells (BMSCs. Materials and Methods: In this experimental study, BMSCs were extracted and expanded to the 3rd passage, then cultured in Dulbecco’s Modified Eagle’s Medium (DMEM complemented with osteogenic media as well as 6 ng/ml and 6 μg/ml of BA. After 5, 10, 15 and 21 days the viability and the level of mineralization was determined using MTT assay and alizarin red respectively. In addition, the morphology, nuclear diameter and cytoplasmic area of the cells were studied with the help of fluorescent dye. The concentration of calcium, activity of alanine transaminase (ALT, aspartate transaminase (AST, lactate dehydrogenase (LDH and alkaline phosphatase (ALP as well as sodium and potassium levels were also evaluated using commercial kits and a flame photometer respectively. Results: Although 6 μg/ml of BA was found to be toxic, a concentration of 6 ng/ml increased the osteogenic ability of the cell significantly throughout the treatment. In addition it was observed that B treatment caused the early induction of matrix mineralization compared to controls. Conclusion: Although more investigation is required, we suggest the prescription of a very low concentration of B in the form of BA or foods containing BA, in groups at high risk of osteoporosis or in the case of bone fracture.

  12. Effect of Chromatin-Remodeling Agents in Hepatic Differentiation of Rat Bone Marrow-Derived Mesenchymal Stem Cells In Vitro and In Vivo

    Directory of Open Access Journals (Sweden)

    Danna Ye

    2016-01-01

    Full Text Available Epigenetic events, including covalent histone modifications and DNA methylation, play fundamental roles in the determination of lineage-specific gene expression and cell fates. The aim of this study was to determine whether the DNA methyltransferase inhibitor (DNMTi 5-aza-2′-deoxycytidine (5-aza-dC and the histone deacetylase inhibitor (HDACi trichostatin A (TSA promote the hepatic differentiation of rat bone marrow-derived mesenchymal stem cells (rBM-MSCs and their therapeutic effect on liver damage. 1 μM TSA and 20 μM 5-aza-dC were added to standard hepatogenic medium especially at differentiation and maturation steps and their potential function on hepatic differentiation in vitro and in vivo was determined. Exposure of rBM-MSCs to 1 μM TSA at both the differentiation and maturation steps considerably improved hepatic differentiation. TSA enhanced the development of the hepatocyte shape, promoted the chronological expression of hepatocyte-specific markers, and improved hepatic functions. In contrast, treatment of rBM-MSCs with 20 μM 5-aza-dC alone or in combination with TSA was ineffective in improving hepatic differentiation in vitro. TSA and/or 5-aza-dC derived hepatocytes-like cells failed to improve the therapeutic potential in liver damage. We conclude that HDACis enhance hepatic differentiation in a time-dependent manner, while DNMTis do not induce the hepatic differentiation of rBM-MSCs in vitro. Their in vivo function needs further investigation.

  13. Opposite effects of bone marrow-derived cells transplantation in MPTP-rat model of Parkinson's disease: a comparison study of mononuclear and mesenchymal stem cells.

    Science.gov (United States)

    Capitelli, Caroline Santos; Lopes, Carolina Salomão; Alves, Angélica Cristina; Barbiero, Janaína; Oliveira, Lucas Felipe; da Silva, Valdo José Dias; Vital, Maria Aparecida Barbato Frazão

    2014-01-01

    The 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) animal model is a useful tool to study Parkinson's disease (PD) and was used in the present study to investigate the potential beneficial as well as deleterious effects of systemic bone-marrow mononuclear cell (BMMC) or mesenchymal stem cell (BM-MSC) transplantation. MPTP administration resulted in a breakdown of the blood-brain barrier and motor impairment in the open field test 24 h after surgery. Three and 7 days after receiving the lesion, the injured animals showed remaining motor impairment compared to the sham groups along with a significant loss of tyrosine hydroxylase-immunoreactive (TH-ir) cells in the substantia nigra pars compacta (SNpc). The MPTP-lesioned rats treated with BMMCs immediately after lesioning exhibited motor impairment similar to the MPTP-saline group, though they presented a significantly higher loss of TH-ir cells in the SNpc compared to the MPTP-saline group. This increased loss of TH-ir cells in the SNpc was not observed when BMMC transplantation was performed 24 h after MPTP administration. In contrast, in the MPTP animals treated early with systemic BM-MSCs, no loss of TH-ir cells was observed. BMMCs and BM-MSCs previously labeled with CM-DiI cell tracker were found in brain sections of all transplanted animals. In addition, cells expressing CD45, an inflammatory white blood cell marker, were found in all brain sections analyzed and were more abundant in the MPTP-BMMC animals. In these animals, Iba1+ microglial cells showed also marked morphological changes indicating increased microglial activation. These results show that systemic BMMC transplantation did not ameliorate or prevent the lesion induced by MPTP. Instead, BMMC transplantation in MPTP-lesioned rats accelerated dopaminergic neuronal damage and induced motor impairment and immobility behavior. These findings suggest that caution should be taken when considering cell therapy using BMMCs to treat PD. However, systemic

  14. Promise(s of mesenchymal stem cells as an in vitro model system to depict pre-diabetic/diabetic milieu in WNIN/GR-Ob mutant rats.

    Directory of Open Access Journals (Sweden)

    Soundarya L Madhira

    Full Text Available BACKGROUND: Development of model systems have helped to a large extent, in bridging gap to understand the mechanism(s of disease including diabetes. Interestingly, WNIN/GR-Ob rats (Mutants, established at National Centre for Laboratory Animals (NCLAS of National Institute of Nutrition (NIN, form a suitable model system to study obesity with Type 2 diabetes (T2D demonstrating several secondary complications (cataract, cardiovascular complications, infertility, nephropathy etc. The present study has been carried out to explore the potent application(s of multipotent stem cells such as bone marrow mesenchymal stem cells (BM-MSCs, to portray features of pre-diabetic/T2D vis-à-vis featuring obesity, with impaired glucose tolerance (IGT, hyperinsulinemia (HI and insulin resistance (IR seen with Mutant rats akin to human situation. METHODOLOGY/PRINCIPAL FINDINGS: Primary cultures of BM-MSCs (third passage from Mutants, its lean littermate (Lean and parental control (Control were characterized for: proliferation markers, disease memory to mark obesity/T2D/HI/IR which included phased gene expression studies for adipogenic/pancreatic lineages, inflammatory markers and differentiation ability to form mature adipocytes/Insulin-like cellular aggregates (ILCAs. The data showed that BM-MSCs from Mutant demonstrated a state of disease memory, depicted by an upregulated expression of inflammatory markers (IL-6, TNFα; increased stem cell recruitment (Oct-4, Sox-2 and proliferation rates (CD90+/CD29+, PDA, 'S' phase of cell cycle by FACS and BrdU incorporation; accelerated preadipocyte induction (Dact-1, PPARγ2 with a quantitative increase in mature adipocyte formation (Leptin; ILCAs, which were non-responsive to high glucose did confer the Obese/T2D memory in Mutants. Further, these observations were in compliance with the anthropometric data. CONCLUSIONS: Given the ease of accessibility and availability of MSCs, the present study form the basis to report for

  15. Mesenchymal stem cells with overexpression of midkine enhance cell survival and attenuate cardiac dysfunction in a rat model of myocardial infarction

    NARCIS (Netherlands)

    S.-L. Zhao (Shu-Li); Y. Zhang (Yaojun); M.-H. Li (Ming-Hui); X.-L. Zhang (Xin-Lei); S.-L. Chen (Shao-Liang)

    2014-01-01

    textabstractIntroduction. Elevated midkine (MK) expression may contribute to ventricular remodeling and ameliorate cardiac dysfunction after myocardial infarction (MI). Ex vivo modification of signaling mechanisms in mesenchymal stem cells (MSCs) with MK overexpression may improve the efficacy of ce

  16. Curative effect of transplantation of mesenchymal stem cells transfected with recombinant lentiviral vectors carrying brain-derived neurotrophic factor gene on intracerebral hemorrhage in rats

    Institute of Scientific and Technical Information of China (English)

    任瑞芳

    2013-01-01

    Objective To observe the curative effect of transplantation of mesenchymal stem cells(MSCs) transfected with recombinant lentiviral vectors carrying brain-derived neurotrophic factor(BDNF) gene on intracerebral

  17. Propofol promotes spinal cord injury repair by bone marrow mesenchymal stem cell transplantation

    Directory of Open Access Journals (Sweden)

    Ya-jing Zhou

    2015-01-01

    Full Text Available Propofol is a neuroprotective anesthetic. Whether propofol can promote spinal cord injury repair by bone marrow mesenchymal stem cells remains poorly understood. We used rats to investigate spinal cord injury repair using bone marrow mesenchymal stem cell transplantation combined with propofol administration via the tail vein. Rat spinal cord injury was clearly alleviated; a large number of newborn non-myelinated and myelinated nerve fibers appeared in the spinal cord, the numbers of CM-Dil-labeled bone marrow mesenchymal stem cells and fluorogold-labeled nerve fibers were increased and hindlimb motor function of spinal cord-injured rats was markedly improved. These improvements were more prominent in rats subjected to bone marrow mesenchymal cell transplantation combined with propofol administration than in rats receiving monotherapy. These results indicate that propofol can enhance the therapeutic effects of bone marrow mesenchymal stem cell transplantation on spinal cord injury in rats.

  18. Propofol promotes spinal cord injury repair by bone marrow mesenchymal stem cell transplantation

    Institute of Scientific and Technical Information of China (English)

    Ya-jing Zhou; Jian-min Liu; Shu-ming Wei; Yun-hao Zhang; Zhen-hua Qu; Shu-bo Chen

    2015-01-01

    Propofol is a neuroprotective anesthetic. Whether propofol can promote spinal cord injury repair by bone marrow mesenchymal stem cells remains poorly understood. We used rats to investigate spinal cord injury repair using bone marrow mesenchymal stem cell transplantation combined with propofol administrationvia the tail vein. Rat spinal cord injury was clearly alleviated; a large number of newborn non-myelinated and myelinated nerve ifbers appeared in the spinal cord, the numbers of CM-Dil-labeled bone marrow mesenchymal stem cells and lfuorogold-labeled nerve ifbers were increased and hindlimb motor function of spinal cord-injured rats was mark-edly improved. These improvements were more prominent in rats subjected to bone marrow mesenchymal cell transplantation combined with propofol administration than in rats receiving monotherapy. These results indicate that propofol can enhance the therapeutic effects of bone marrow mesenchymal stem cell transplantation on spinal cord injury in rats.

  19. Quantum measurement corrections to chemically induced dynamic nuclear polarization

    CERN Document Server

    Kominis, I K

    2013-01-01

    Chemically induced dynamic nuclear polarization has emerged as a universal signature of spin order in photosynthetic reaction centers. Such polarization, significantly enhanced above thermal equilibrium, is known to result from the nuclear spin sorting inherent in the radical pair mechanism underlying long-lived charge-separated states in photosynthetic reaction centers. We will here show that the recently understood fundamental quantum dynamics of radical-ion-pair reactions open up a new and completely unexpected venue towards obtaining CIDNP signals. The fundamental decoherence mechanism inherent in the recombination process of radical pairs is shown to produce nuclear spin polarizations on the order of $10^4$ times or more higher than thermal equilibrium values at low fields relevant to natural photosynthesis in earth's magnetic field. This opens up the possibility of a fundamentally new exploration of the biological significance of high nuclear polarizations in photosynthesis.

  20. Combined transplantation of bone marrow mesenchymal stem cells and pedicled greater omentum promotes locomotor function and regeneration of axons after spinal cord injury in rats

    Institute of Scientific and Technical Information of China (English)

    Liang Li; Zhiying Zhang; Haiyan Lin; Congli Ren; Chuansen Zhang

    2008-01-01

    BACKGROUND: According to previous studies, the neuroprotective effect of the pedicled greater omentum may be attributed to the secretion of neurotrophic factors and stimulation of angiogenesis. The neurotrophic factors released from the pedicled greater omentum, such as brain-derived neurotrophic factor and neurotrophin 3/4/5 could exert a neuroprotective effect on the damaged host neural and glial cells, and also could induce the transdifferentiation of transplanted bone marrow mesenchymal stem cells (BMSCs) into neural cells. OBJECTIVE: Based on the functions of the omentum of neuro-protection and vascularization, we hypothesize that the transplantation of BMSCs and pedicled greater omentum into injured rat spinal cord might improve the survival rate and neural differentiation of transplanted BMSCs and consequently gain a better functional outcome. DESIGN, TIME AND SETTING: A randomized, controlled animal experiment. The experiments were carried out at the Department of Anatomy, the Secondary Military Medical University of Chinese PLA between June 2005 and June 2007.MATERIALS: Fifteen male inbred Wistar rats, weighing (200±20) g, provided by the Experimental Animal Center of the Secondary Military Medical University of Chinese PLA were used and met the animal ethical standards. Mouse anti-BrdU and mouse anti-NF200 monoclonal antibody were purchased from Boster, China. METHODS: Cell culture: We used inbred Sprague-Dawley rats to harvest bone marrow for culture of BMSCs and transplantation to avoid possible immune rejection. BMSCs were cultured via total bone marrow adherence. Experimental grouping and intervention: The rats were randomly divided into a control group, cell group and combined group, five rats per group. Rats in the control group underwent spinal cord injury (SCI) only, during which an artery clamp with pressure force of 30 g was employed to compress the spinal cord at the T10 level for 30 seconds to produce the SCI model. 5 μL PBS containing 105

  1. Immunosuppressive effects of rat mesenchymal stem cells:involvement of CD4+CD25+regulator y T cells

    Institute of Scientific and Technical Information of China (English)

    Zhou Ye; Yan Wang; Hai-Yang Xie and Shu-Sen Zheng

    2008-01-01

    BACKGROUND: Recent studies show that mesenchymal stem cells (MSCs) have immunomodulatory properties. They suppress the immune response to alloantigen and modify the proliferation of T cells. CD4+CD25+ regulatory T cells have strong immunomodulatory potential. However, little is known about the effects of rat MSCs (rMSCs) on the development of regulatory T cells. METHODS:MSCs were obtained from bone marrow of male Sprague-Dawley rats, and co-cultured with CD3+ T cells from allogeneic spleen cells. The proportion of CD4+CD25+ regulatory  T  cells  was  analyzed  by  lfow  cytometry.  To further conifrm the immunosuppressive activity of rMSCs, we used MTT assay and lfow cytometry of CD3+ T cells to investigate the proliferative responses of CD3+ T cells to mitogenic stimuli. Enzyme-linked immunosorbent assay was performed to detect alterations of the cytokines TNF-α, TGF-β and IL-10. RESULTS:The proliferation of CD3+ T cells decreased when co-cultured with rMSCs, and the degree of inhibition was concentration-dependent. The percentage of CD4+CD25+ regulatory T cells increased when CD3+ T cells were co-cultured with different concentrations of rMSCs. The levels of pro-inlfammatory cytokine (TNF-α) decreased while anti-inlfammatory (TGF-β, IL-10) cytokines increased in mixed lymphocyte reaction. CONCLUSIONS:rMSCs inhibit allogeneic T cell proliferation in mixed cell cultures. This immunosuppressive effect seems  to  be  mediated  by  inducing  the  generation  of CD4+CD25+ regulatory T cells and soluble factors.

  2. 大鼠骨髓间充质干细胞的培养与鉴定%Culture and identification of rat bone marrow mesenchymal stem cells

    Institute of Scientific and Technical Information of China (English)

    李晓峰; 赵劲民; 苏伟; 崔向荣; 罗世兴; 马爱国

    2011-01-01

    BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are not rich in bone marrow, and are difficult to culture in vitro.It is crucial to tissue engineering and to the cell experiments in vivo or in vitro that isolating and culturing in vitro BMSCs with high purity, strong vitality and homogeneous biological characteristics.OBJECTIVE: To establish a method of isolation, cultivation and purification of rat BMSCs in vitro, to observe cell morphology,and to assess surface markers and multi-directional differentiation capacity.METHODS: BMSCs from rats were isolated, cultured and purified by the whole bone marrow adherence method, for morphology observations, the growth curve was drawn, cell cycle was analyzed, cell surface markers were assessed by flow cytometry, and BMSCs were induced to differentiate into osteoblasts and adipocytes.RESULTS AND CONCLUSION : BMSCs from rats were spindle cell-based, showing radial colony arrangement. Cells kept strong growth and could passage in continuous and stable manner over 10 passages. The growth curve and cell cycle demonstrated that BMSCs were consistent with the growth characteristics and good activity of normal cells. At the third passage, BMSCs were negative for CD34 and CD45, but positive for CD44, CD90 and CD105. Following induction, oil red O staining, alkaline phosphatase staining, von Kossa staining and alizarin red staining produced a strong reaction in cells. Whole bone marrow adherence method is simple and can isolate, purify and amplify BMSCs in vitro. The obtained cells have general biological characteristics of mesenchymal stem cells, and also have potentiality of multi-directional differentiation. This experimental method has important practical significance to provide adequate source of seed cells for tissue engineering.%背景:骨髓间充质干细胞在骨髓中含量极低,体外培养难度较大.体外分离培养纯度高、活力强、生物特性均一的间充质干细胞,对组织工程及细胞的体

  3. Effects of nerve growth factor and basic fibroblast growth factor dual gene modification on rat bone marrow mesenchymal stem cell differentiation into neuron-like cells in vitro.

    Science.gov (United States)

    Hu, Yang; Zhang, Yan; Tian, Kang; Xun, Chong; Wang, Shouyu; Lv, Decheng

    2016-01-01

    Recent studies regarding regenerative medicine have focused on bone marrow mesenchymal stem cells (BMSCs), which have the potential to undergo neural differentiation, and may be transfected with specific genes. BMSCs can differentiate into neuron‑like cells in certain neurotropic circumstances in vitro. Basic fibroblast growth factor (bFGF) and nerve growth factor (NGF) are often used to induce neural differentiation in BMSCs in vitro. However, previous studies regarding their combined actions are insufficient. The present study is the first, to the best of our knowledge, to thoroughly assess the enhancement of neural differentiation of BMSCs following transfection with bFGF and NGF. Sprague‑Dawley (SD) rat BMSCs were separated through whole bone marrow adherence, and were then passaged to the third generation. The cells were subsequently divided into five groups: The control group, which consisted of untransfected BMSCs; the plv‑blank‑transfected BMSCs group; the plv‑bFGF‑transfected BMSCs group; the plv‑NGF‑transfected BMSCs group; and the plv‑NGF‑bFGF co‑transfected BMSCs group. Cell neural differentiation was characterized in terms of stem cell molecular expression, and the neuronal morphology and expression of neural‑like molecules was detected in each of the groups. A total of 72 h post‑transfection, the expression levels of neuron‑specific enolase, glial fibrillary acidic protein, and nestin protein, were higher in the co‑transfected group, as compared with the other groups, the expression levels of β‑tubulin III were also increased in the co‑transfected cells, thus suggesting the maturation of differentiated neuron‑like cells. Furthermore, higher neuronal proliferation was observed in the co‑transfected group, as compared with the other groups at passages 2, 4, 6 and 8. Western blotting demonstrated that the transfected groups exhibited a simultaneous increase in phosphorylation of the AKT and extracellular signal

  4. Improved dopamine transporter binding activity after bone marrow mesenchymal stem cell transplantation in a rat model of Parkinson's disease: small animal positron emission tomography study with F-18 FP-CIT

    Energy Technology Data Exchange (ETDEWEB)

    Park, Bok-Nam; Lee, Kwanjae; An, Young-Sil [School of Medicine, Ajou University, Department of Nuclear Medicine and Molecular Imaging, Woncheon-dong, Yeongtong-gu, Gyeonggi-do, Suwon (Korea, Republic of); Kim, Jang-Hee; Park, So Hyun [Ajou University School of Medicine, Department of Pathology, Suwon (Korea, Republic of)

    2015-05-01

    We evaluated the effects of bone marrow-derived mesenchymal stem cells (BMSCs) in a model of Parkinson's disease (PD) using serial F-18 fluoropropylcarbomethoxyiodophenylnortropane (FP-CIT) PET. Hemiparkinsonian rats were treated with intravenously injected BMSCs, and animals without stem cell therapy were used as the controls. Serial FP-CIT PET was performed after therapy. The ratio of FP-CIT uptake in the lesion side to uptake in the normal side was measured. The changes in FP-CIT uptake were also analyzed using SPM. Behavioural and histological changes were observed using the rotational test and tyrosine hydroxylase (TH)-reactive cells. FP-CIT uptake ratio was significantly different in the BMSCs treated group (n = 28) at each time point. In contrast, there was no difference in the ratio in control rats (n = 25) at any time point. SPM analysis also revealed that dopamine transporter binding activity was enhanced in the right basal ganglia area in only the BMSC therapy group. In addition, rats that received BMSC therapy also exhibited significantly improved rotational behaviour and preservation of TH-positive neurons compared to controls. The therapeutic effect of intravenously injected BMSCs in a rat model of PD was confirmed by dopamine transporter PET imaging, rotational functional studies, and histopathological evaluation. (orig.)

  5. Protein's native state stability in a chemically induced denaturation mechanism.

    Science.gov (United States)

    Olivares-Quiroz, L; Garcia-Colin, L S

    2007-05-21

    In this work, we present a generalization of Zwanzig's protein unfolding analysis [Zwanzig, R., 1997. Two-state models of protein folding kinetics. Proc. Natl Acad. Sci. USA 94, 148-150; Zwanzig, R., 1995. Simple model of protein folding kinetics. Proc. Natl Acad. Sci. USA 92, 9801], in order to calculate the free energy change Delta(N)(D)F between the protein's native state N and its unfolded state D in a chemically induced denaturation. This Extended Zwanzig Model (EZM) is both based on an equilibrium statistical mechanics approach and the inclusion of experimental denaturation curves. It enables us to construct a suitable partition function Z and to derive an analytical formula for Delta(N)(D)F in terms of the number K of residues of the macromolecule, the average number nu of accessible states for each single amino acid and the concentration C(1/2) where the midpoint of the ND transition occurs. The results of the EZM for proteins where chemical denaturation follows a sigmoidal-type profile, as it occurs for the case of the T70N human variant of lysozyme (PDB code: T70N) [Esposito, G., et al., 2003. J. Biol. Chem. 278, 25910-25918], can be splitted into two lines. First, EZM shows that for sigmoidal denaturation profiles, the internal degrees of freedom of the chain play an outstanding role in the stability of the native state. On the other hand, that under certain conditions DeltaF can be written as a quadratic polynomial on concentration C(1/2), i.e., DeltaF approximately aC(1/2)(2)+bC(1/2)+c, where a,b,c are constant coefficients directly linked to protein's size K and the averaged number of non-native conformations nu. Such functional form for DeltaF has been widely known to fit experimental measures in chemically induced protein denaturation [Yagi, M., et al., 2003. J. Biol. Chem. 278, 47009-47015; Asgeirsson, B., Guojonsdottir, K., 2006. Biochim. Biophys. Acta 1764, 190-198; Sharma, S., et al., 2006. Protein Pept. Lett. 13(4), 323-329; Salem, M., et al

  6. Protective effect of bone marrow-derived mesenchymal stem cells on dopaminergic neurons against 1-methyl-4-phenylpyridinium ion-induced neurotoxicity in rat brain slices

    Institute of Scientific and Technical Information of China (English)

    Lirong Jin; Zhen Hong; Chunjiu Zhong; Yang Wang

    2009-01-01

    BACKGROUND: To date, the use of bone marrow-derived mesenchymal stem cells (MSCs) for the treatment of Parkinson's disease have solely focused on in vivo animal models. Because of the number of influencing factors, it has been difficult to determine a consistent outcome. OBJECTIVE: To establish an injury model in brain slices of substantia nigra and striatum using 1-methyl-4-phenylpytidinium ion (MPP+), and to investigate the effect of MSCs on dopaminergic neurons following MPP+ induced damage.DESIGN, TIME AND SETTING: An in vitro, randomized, controlled, animal experiment using immunohistochemistry was performed at the Laboratory of the Department of Anatomy, Fudan University between January 2004 and December 2006.MATERIALS: Primary MSC cultures were obtained from femurs and tibias of adult Sprague Dawley rats. Organotypic brain slices were isolated from substantia nigra and striatum of 1-day-old Sprague Dawley rat pups. Monoclonal antibodies for tyrosine hydroxylase (TH, 1:5 000) were from Santa Cruz (USA); goat anti-rabbit IgG antibodies labeled with FITC were from Boster Company (China).METHODS: Organotypic brain slices were cultured for 5 days in whole culture medium supplemented with 50% DMEM, 25% equine serum, and 25% Tyrode's balanced salt solution. The medium was supplemented with 5 μg/mL Ara-C, and the culture was continued for an additional 5 days. The undergrowth of brain slices was discarded at day 10. Eugonic brain slices were cultured with basal media for an additional 7 days. The brain slices were divided into three groups: control, MPP+ exposure, and co-culture. For the MPP+ group, MPP+ (30 μmol/L) was added to the media at day 17 and brain slices were cultured for 4 days, followed by control media. For the co-culture group, the MPP+ injured brain slices were placed over MSCs in the well and were further cultured for 7 days.MAIN OUTCOME MEASURES: After 28 days in culture, neurite outgrowth was examined in the brain slices under phase

  7. Chemically Induced and Light-Independent Cryptochrome Photoreceptor Activation

    Institute of Scientific and Technical Information of China (English)

    Gesa Rosenfeldt; Rafael Mu(n)oz Viana; Henning D.Mootz; Albrecht G.Von Arnim; Alfred Batschauer

    2008-01-01

    The cryptochrome photoreceptors of higher plants are dimeric proteins. Their N-terminal photosensory domain mediates dimerization, and the unique C-terminal extension (CCT) mediates signaling. We made use of the human FK506-binding protein (FKBP) that binds with high affinity to rapamycin or rapamycin analogs (rapalogs). The FKBP-rapamycin complex is recognized by another protein, FRB, thus allowing rapamycin-induced dimerization of two target proteins. Here we demonstrate by bioluminescence resonance energy transfer (BRET) assays the applicability of this regulated dimerization system to plants. Furthermore, we show that fusion proteins consisting of the C-terminal domain of Arabidopsis cryptochrome 2 fused to FKBP and FRB and coexpressed in Arabidopsis cells specifically induce the expression of cryptochrome-controlled reporter and endogenous genes in darkness upon incubation with the rapalog. These results demonstrate that the activation of cryptochrome signal transduction can be chemically induced in a dose-dependent fashion and uncoupled from the light signal, and provide the groundwork for gain-of-function experiments to study specifically the role of photoreceptors in darkness or in signaling cross-talk even under light conditions that activate members of all photoreceptor families.

  8. Umbilical cord mesenchymal stem cell transplantation for the treatment of diabetes mellitus in rats%脐带间充质干细胞移植治疗大鼠糖尿病

    Institute of Scientific and Technical Information of China (English)

    李丽; 赵博

    2014-01-01

    背景:脐血间充质干细胞具有很强的增殖能力和分化能力,在趋向分化作用下可以分化成胰岛β细胞,进而起到治疗糖尿病的作用。  目的:观察移植脐带间充质干细胞对大鼠糖尿病的治疗效果。  方法:30只雄性SD大鼠中随机取6只作为对照组,注射生理盐水;其中24只按45 mg/kg的剂量注射链脲霉素建立糖尿病模型后,随机等分为移植组和糖尿病组,移植组大鼠尾静脉注射移植脐带间充质干细胞。  结果与结论:造模后30 d,糖尿病组大鼠空腹血糖维持在较高水平,且高于对照组(P 0.05),而糖尿病组大鼠空腹血糖维持较高水平,且体质量持续下降。提示脐带间充质干细胞移植能有效治疗大鼠糖尿病。%BACKGROUND:Umbilical cord mesenchymal stem cels have strong proliferation and differentiation capacities, and can be induced to differentiate into pancreatic β cels, thereby playing a therapeutic effect on diabetes mel itus. OBJECTIVE:To study the therapeutic effects of transplantation of umbilical cord mesenchymal stem cels for treatment of diabetes melitus in rats. METHODS: Thirty male Sprague-Dawley rats were randomly divided into control group (n=6), transplantation group (n=12) and diabetic group (n=12). Rats in the control group were given normal saline injection. Rats in the other two groups were injected with streptozotocin at a dose of 45 mg/kg to establish diabetic models. After modeling, transplantation of umbilical cord mesenchymal stem celsviatail vein was given in the transplantation group. RESULTS AND CONCLUSION:Thirty days after modeling, the fasting blood glucose was maintained at a higher level in comparison with the control group (P 0.05), but in the diabetic group, the fasting blood glucose level was stil higher and the body mass continued to decrease. These findings indicate that the transplantation of umbilical cord mesenchymal stem cels can be effective

  9. Evaluation of early healing events around mesenchymal stem cell-seeded collagen-glycosaminoglycan scaffold. An experimental study in Wistar rats.

    LENUS (Irish Health Repository)

    Alhag, Mohamed

    2011-03-01

    Tissue engineering using cell-seeded biodegradable scaffolds offers a new bone regenerative approach that might circumvent many of the limitations of current therapeutic modalities. The aim of this experiment was to study the early healing events around mesenchymal stem cell-seeded collagen-glycosaminoglycan scaffolds.

  10. Rat reverse genetics : generation and characterization of chemically induced rat mutants

    NARCIS (Netherlands)

    van Boxtel, R.

    2010-01-01

    The use of animal models has been crucial for studying the function of genetic elements in the human genome. Embryonic stem (ES) cell-based homologous recombination (HR) has proven a very efficient technique for gene manipulation. However, this technique is not (yet) available for all model organism

  11. A high-throughput chemically induced inflammation assay in zebrafish

    Directory of Open Access Journals (Sweden)

    Liebel Urban

    2010-12-01

    Full Text Available Abstract Background Studies on innate immunity have benefited from the introduction of zebrafish as a model system. Transgenic fish expressing fluorescent proteins in leukocyte populations allow direct, quantitative visualization of an inflammatory response in vivo. It has been proposed that this animal model can be used for high-throughput screens aimed at the identification of novel immunomodulatory lead compounds. However, current assays require invasive manipulation of fish individually, thus preventing high-content screening. Results Here we show that specific, noninvasive damage to lateral line neuromast cells can induce a robust acute inflammatory response. Exposure of fish larvae to sublethal concentrations of copper sulfate selectively damages the sensory hair cell population inducing infiltration of leukocytes to neuromasts within 20 minutes. Inflammation can be assayed in real time using transgenic fish expressing fluorescent proteins in leukocytes or by histochemical assays in fixed larvae. We demonstrate the usefulness of this method for chemical and genetic screens to detect the effect of immunomodulatory compounds and mutations affecting the leukocyte response. Moreover, we transformed the assay into a high-throughput screening method by using a customized automated imaging and processing system that quantifies the magnitude of the inflammatory reaction. Conclusions This approach allows rapid screening of thousands of compounds or mutagenized zebrafish for effects on inflammation and enables the identification of novel players in the regulation of innate immunity and potential lead compounds toward new immunomodulatory therapies. We have called this method the chemically induced inflammation assay, or ChIn assay. See Commentary article: http://www.biomedcentral.com/1741-7007/8/148.

  12. Differential Gene Expression in Chemically Induced Mouse Lung Adenomas

    Directory of Open Access Journals (Sweden)

    Ruisheng Yao

    2003-01-01

    Full Text Available Because of similarities in histopathology and tumor progression stages between mouse and human lung adenocarcinomas, the mouse lung tumor model with lung adenomas as the endpoint has been used extensively to evaluate the efficacy of putative lung cancer chemopreventive agents. In this study, a competitive cDNA library screening (CCLS was employed to determine changes in the expression of mRNA in chemically induced lung adenomas compared with paired normal lung tissues. A total of 2555 clones having altered expression in tumors were observed following competitive hybridization between normal lung and lung adenomas after primary screening of over 160,000 clones from a mouse lung cDNA library. Among the 755 clones confirmed by dot blot hybridization, 240 clones were underexpressed, whereas 515 clones were overexpressed in tumors. Sixty-five clones with the most frequently altered expression in six individual tumors were confirmed by semiquantitative RT-PCR. When examining the 58 known genes, 39 clones had increased expression and 19 had decreased expression, whereas the 7 novel genes showed overexpression. A high percentage (>60% of overexpressed or underexpressed genes was observed in at least two or three of the lesions. Reproducibly overexpressed genes included ERK-1, JAK-1, surfactant proteins A, B, and C, NFAT1, α-1 protease inhibitor, helix-loop-helix ubiquitous kinase (CHUK, α-adaptin, α-1 PI2, thioether S-methyltransferase, and CYP2C40. Reproducibly underexpressed genes included paroxanase, ALDH II, CC10, von Ebner salivary gland protein, and α- and β-globin. In addition, CCLS identified several novel genes or genes not previously associated with lung carcinogenesis, including a hypothetical protein (FLJ11240 and a guanine nucleotide exchange factor homologue. This study shows the efficacy of this methodology for identifying genes with altered expression. These genes may prove to be helpful in our understanding of the genetic basis of

  13. 骨髓间充质干细胞移植对急性期脑梗死大鼠的干预作用%The effect of bone marrow mesenchymal stem cell transplantation on cerebral infarction in rats

    Institute of Scientific and Technical Information of China (English)

    赵小妹; 刘永刚; 许云鹤; 曹姍; 赵静; 张祥建

    2012-01-01

    目的 观察骨髓间充质干细胞(BMSCs)移植对急性期脑梗死大鼠的干预作用.方法 按随机化原则将50只脑梗死大鼠模型随机分为BMSCs移植组、上清液移植组各22只及对照组6只.BMSCs组在脑梗死大鼠模型成功后3 h经静脉注入BMSCs 1 ml,上清液组注入上清液1 ml,对照组注入磷酸盐缓冲液(PBS)1 ml.术后28 d检测PBMSc在脑内的存活转化情况.分别于0、7、14及28 d观察记录各组大鼠的神经损伤严重程度评分(NSS).结果 28 d后,BMSCs脑梗死灶边缘可见少量BrdU阳性细胞表达神经元特异性烯醇化酶(NSE),而上清液组及对照组病灶边缘则无NSE;细胞移植后,各组神经功能均有不同程度的恢复;从第7天起,BMSCs移植组NSS评分明显低于上清液组及对照组(P0.05).结论 骨髓间充质干细胞移植入急性脑梗死大鼠体内后,可存活、移行至梗死灶周围,并分化为神经元细胞,而且能够减轻脑梗死大鼠的神经损伤程度.%Objective To observe the effect of transplanted bone marrow mesenchymal stem cells on the brain of rats with cerebral infarction. Methods Fifty middle cerebral artery occlusion model were randomly divided into control group ( n = 6 ), group with supernatant transplantation ( n = 22 ) and group with BMSCs transplantation ( n =22 ). 3h after animal model was established, PBS, supernatant and BMSCs was injected intravenously in three groups respectively. 28d after transplantation, the survival condition of transplanted bone marrow mesenchymal stem cells in the brain of rats with cerebral infarction were observed, the expression of neuron specific enolase in brain tissue and the neurological severity score ( NSS ) were compared among all the groups 0,7,14 and 28d after operation. Results At day 28 after bone marrow mesenchymal stem cell transplantation, under fluorescence microscope there were a large quantity of BrdU positive cells aggregated in the marginal zone of cerebral infarction

  14. Polyurethane/polylactide-based biomaterials combined with rat olfactory bulb-derived glial cells and adipose-derived mesenchymal stromal cells for neural regenerative medicine applications

    Energy Technology Data Exchange (ETDEWEB)

    Grzesiak, Jakub, E-mail: grzesiak.kuba@gmail.com [Electron Microscopy Laboratory, University of Environmental and Life Sciences, Kozuchowska 5b, 51-631 Wroclaw (Poland); Marycz, Krzysztof [Electron Microscopy Laboratory, University of Environmental and Life Sciences, Kozuchowska 5b, 51-631 Wroclaw (Poland); Szarek, Dariusz [Department of Neurosurgery, Lower Silesia Specialist Hospital of T. Marciniak, Emergency Medicine Center, Traugutta 116, 50-420 Wroclaw (Poland); Bednarz, Paulina [State Higher Vocational School in Tarnów, Mickiewicza 8, 33-100 Tarnów (Poland); Laska, Jadwiga [AGH University of Science and Technology, Faculty of Materials Science and Ceramics, Mickiewicza 30, 30-059 Kraków (Poland)

    2015-07-01

    Research concerning the elaboration and application of biomaterial which may support the nerve tissue regeneration is currently one of the most promising directions. Biocompatible polymer devices are noteworthy group among the numerous types of potentially attractive biomaterials for regenerative medicine application. Polylactides and polyurethanes may be utilized for developing devices for supporting the nerve regeneration, like nerve guide conduits or bridges connecting the endings of broken nerve tracts. Moreover, the combination of these biomaterial devices with regenerative cell populations, like stem or precursor cells should significantly improve the final therapeutic effect. Therefore, the composition and structure of final device should support the proper adhesion and growth of cells destined for clinical application. In current research, the three polymer mats elaborated for connecting the broken nerve tracts, made from polylactide, polyurethane and their blend were evaluated both for physical properties and in vitro, using the olfactory-bulb glial cells and mesenchymal stem cells. The evaluation of Young's modulus, wettability and roughness of obtained materials showed the differences between analyzed samples. The analysis of cell adhesion, proliferation and morphology showed that the polyurethane–polylactide blend was the most neutral for cells in culture, while in the pure polymer samples there were significant alterations observed. Our results indicated that polyurethane–polylactide blend is an optimal composition for culturing and delivery of glial and mesenchymal stem cells. - Highlights: • Polyurethane–polylactide blends exhibit different characteristics from pure polymers. • Pure PU and PLA negatively influence on morphology of glial and mesenchymal cells. • PU/PLA blend was neutral for glial and mesenchymal cell proliferation and morphology.

  15. CM-Dil标记大鼠骨髓间充质干细胞的生物学特性探讨%Effect of CM-Dil labeling on biological characteristics of rat bone marrow mesenchymal stem cells

    Institute of Scientific and Technical Information of China (English)

    张雅妮; 于美娟; 冯善伟; 王淑辉; 李美山; 熊符; 张成

    2016-01-01

    背景:目前已有部分实验采用CM-Dil标记间充质干细胞开展体内外实验,取得良好成果。  目的:观察CM-Dil标记大鼠骨髓间充质干细胞的生物学特性及其定向迁移能力。  方法:分离培养大鼠骨髓间充质干细胞,通过流式细胞技术分析其特异性细胞表面标记抗原(CD29, CD44,CD11b,CD45)的表达。用CM-Dil标记大鼠骨髓间充质干细胞,继续培养细胞,观察细胞的形态及增殖情况,及传代后荧光的强度与持久性。采用Transwell小室进行标记后大鼠骨髓间充质干细胞向mdx鼠腓肠肌匀浆液的迁移实验。  结果与结论:所培养细胞均一性好,均表达CD29和CD44,不表达CD11b和CD45,符合骨髓间充质干细胞的特点。经CM-Dil标记后对大鼠骨髓间充质干细胞的形态、增殖无明显影响,传代后仍可保持荧光强度,但随传代有所减弱。标记前后大鼠骨髓间充质干细胞的迁移能力无显著性差异(P>0.05)。结果说明,CM-Dil标记过程安全、简单、高效,对大鼠骨髓间充质干细胞的形态、增殖及迁移能力无明显影响,体外细胞传代后仍可保持荧光,是大鼠骨髓间充质干细胞标记与示踪的理想试剂。%BACKGROUND:Existing evidence has shown that CM-Dil-labeled mesenchymal stem cel s have delivered better results in experiments in vivo and in vitro. OBJECTIVE:To investigate the effect of in vitro CM-Dil labeling on biological characteristics and directed migration of rat bone marrow mesenchymal stem cel s. METHODS:Mesenchymal stem cel s were isolated and cultured from the bone marrow of Sprague-Dawley rats. The specific surface antigens, CD29, CD44, CD11b, CD45, were detected by flow cytometry. The rat bone marrow mesenchymal stem cel s were labeled with CM-Dil, and then were cultured and passed continual y. Thereafter the morphology and proliferation ability of labeled cel s were assessed. The strength and

  16. 脐血干细胞移植对帕金森病大鼠旋转行为的影响%Effect of human umbilical cord blood mesenchymal stem cells transplantation on rotational behavior of Parkinson's disease rats

    Institute of Scientific and Technical Information of China (English)

    樊志刚; 刘芳

    2012-01-01

    背景:目前帕金森病的临床治疗还是以药物为主,细胞移植实验也多见于骨髓间充质干细胞,脐血来源干细胞移植能否改善帕金森病的旋转行为报道较少.目的:观察脐血间充质干细胞移植对帕金森病大鼠旋转行为的影响.方法:帕金森病模型大鼠随机分成实验组和对照组.实验组大鼠纹状体内植入用Hoechst33258标记的第4代脐血间充质干细胞,对照组注射PBS.此后每周腹腔注射阿扑吗啡以观察大鼠的旋转行为;并在移植后3,6,9周用免疫荧光双标法检测间充质干细胞的存活、迁移情况以及胶质纤维酸性蛋白、神经元特异性烯醇化酶、酪氨酸羟化酶和突触素的表达.结果与结论:移植脐血间充质干细胞后大鼠的旋转行为与对照组相比有明显改善(P < 0.05);间充质干细胞可在大鼠脑内存活,随时间延长迁移范围扩大,分布于纹状体、胼胝体和皮质;胶质纤维酸性蛋白、神经元特异性烯醇化酶、酪氨酸羟化酶都有表达,突触素无表达.结果可见移植脐血间充质干细胞后能明显改善帕金森病大鼠旋转行为,有望成为治疗帕金森病的种子细胞.%BACKGROUND: To date, the clinical treatment of Parkinson's disease (PD) mainly depends on drug, and as for celltransplantation experiment, bone marrow mesenchymal stem cells (BMSCs) transplantation is the common method. The reportsabout whether umbilical cord blood mesenchymal stem cells (UCBMSCs) transplantation can improve the rotational behavior arerare.OBJECTIVE: To explore the effect of human UCBMSCs transplantation on rotational behavior of PD rats.METHODS: The PD rat models were divided into the experimental group (n=20) and the control group (n=20). The fourthgeneration of MSCs were marked by Hoechst33258 and then transplanted into rat striatum in experimental group, and the rats incontrol group were given PBS. Apomorphine was injected intraperitoneally to examine the rotational

  17. 人胎盘间充质干细胞移植修复大鼠卵巢早衰的实验研究%Human Placental Mesenchymal Stem Cells Transplantation for the Repair of rat Premature Ovarian Failure

    Institute of Scientific and Technical Information of China (English)

    李永丽; 王永峰; 相丽; 徐仙

    2016-01-01

    目的:为探讨尾静脉注射移植人胎盘间充质干细胞(hpMSCs)对化疗导致的卵巢早衰模型大鼠卵巢功能的修复作用。方法经鉴定的人胎盘间充质干细胞经尾静脉注射移植给卵巢早衰模型大鼠,实验分为hpMSCs组和干细胞培养液移植组(control组),每组10只,分别通过尾静脉等体积注射,注射后观察大鼠卵巢结构损伤情况,比较血清性激素(E2、AMH、 INHB)浓度。结果大鼠卵巢激素水平与control组相比,hpMSCs移植前后血清E2和INHB水平有统计学差异;血清AMH水平无统计学差异(P=0.051);大鼠卵巢组织形态学结果表明,与模型组大鼠卵巢皮质损伤严重,尾静脉注射hpMSCs后Bcl-2表达显著降低。结论 hAMSCs大鼠尾静脉移植改善激素水平,减少卵巢细胞凋亡,从而修复卵巢功能。%Objective To explore intravenous transplantation of human placenta derived mesenchymal stem cells (hpMSCs) on repair of chemotherapy-induced ovarian function in a rat model of premature ovarian failure.Method The identiifcation of human placenta derived mesenchymal stem cells transplantation by intravenous injection to rat model of premature ovarian failure, the experiments were divided into hpMSCs group and stem cell culture medium transplantation group (control group), each Group 10, respectively, by the tail vein and other volume injection, after injection, the rats were observed in the ovary structure damage, and compared serum sex hormone (E2, AMH, INHB) concentration.Result Rats Compared with the control group, the levels of serum E2 and INHB were statistically different before and after hpMSCs transplantation. Serum AMH levels were not statistically signiifcant (P=0.051). The results of histological changes in the ovaries of rats showed that the expression of hpMSCs was signiifcantly decreased in the rats with the model group, and the expression of Bcl-2 was signiifcantly decreased after the injection

  18. Intravenous transplantation of bone marrow mesenchymal stem cells promotes neural regeneration after traumatic brain injury

    Institute of Scientific and Technical Information of China (English)

    Fatemeh Anbari; Mohammad Ali Khalili; Ahmad Reza Bahrami; Arezoo Khoradmehr; Fatemeh Sadeghian; Farzaneh Fesahat; Ali Nabi

    2014-01-01

    To investigate the supplement of lost nerve cells in rats with traumatic brain injury by intrave-nous administration of allogenic bone marrow mesenchymal stem cells, this study established a Wistar rat model of traumatic brain injury by weight drop impact acceleration method and ad-ministered 3 × 106 rat bone marrow mesenchymal stem cells via the lateral tail vein. At 14 days after cell transplantation, bone marrow mesenchymal stem cells differentiated into neurons and astrocytes in injured rat cerebral cortex and rat neurological function was improved significant-ly. These findings suggest that intravenously administered bone marrow mesenchymal stem cells can promote nerve cell regeneration in injured cerebral cortex, which supplement the lost nerve cells.

  19. CD-REST: a system for extracting chemical-induced disease relation in literature

    OpenAIRE

    Xu, Jun; Wu, Yonghui; Zhang, Yaoyun; Wang, Jingqi; Lee, Hee-jin; Xu, Hua

    2016-01-01

    Mining chemical-induced disease relations embedded in the vast biomedical literature could facilitate a wide range of computational biomedical applications, such as pharmacovigilance. The BioCreative V organized a Chemical Disease Relation (CDR) Track regarding chemical-induced disease relation extraction from biomedical literature in 2015. We participated in all subtasks of this challenge. In this article, we present our participation system Chemical Disease Relation Extraction SysTem (CD-RE...

  20. Characterization of chemically induced liver injuries using gene co-expression modules.

    Directory of Open Access Journals (Sweden)

    Gregory J Tawa

    Full Text Available Liver injuries due to ingestion or exposure to chemicals and industrial toxicants pose a serious health risk that may be hard to assess due to a lack of non-invasive diagnostic tests. Mapping chemical injuries to organ-specific damage and clinical outcomes via biomarkers or biomarker panels will provide the foundation for highly specific and robust diagnostic tests. Here, we have used DrugMatrix, a toxicogenomics database containing organ-specific gene expression data matched to dose-dependent chemical exposures and adverse clinical pathology assessments in Sprague Dawley rats, to identify groups of co-expressed genes (modules specific to injury endpoints in the liver. We identified 78 such gene co-expression modules associated with 25 diverse injury endpoints categorized from clinical pathology, organ weight changes, and histopathology. Using gene expression data associated with an injury condition, we showed that these modules exhibited different patterns of activation characteristic of each injury. We further showed that specific module genes mapped to 1 known biochemical pathways associated with liver injuries and 2 clinically used diagnostic tests for liver fibrosis. As such, the gene modules have characteristics of both generalized and specific toxic response pathways. Using these results, we proposed three gene signature sets characteristic of liver fibrosis, steatosis, and general liver injury based on genes from the co-expression modules. Out of all 92 identified genes, 18 (20% genes have well-documented relationships with liver disease, whereas the rest are novel and have not previously been associated with liver disease. In conclusion, identifying gene co-expression modules associated with chemically induced liver injuries aids in generating testable hypotheses and has the potential to identify putative biomarkers of adverse health effects.

  1. Characterization of Chemically Induced Liver Injuries Using Gene Co-Expression Modules

    Science.gov (United States)

    Tawa, Gregory J.; AbdulHameed, Mohamed Diwan M.; Yu, Xueping; Kumar, Kamal; Ippolito, Danielle L.; Lewis, John A.; Stallings, Jonathan D.; Wallqvist, Anders

    2014-01-01

    Liver injuries due to ingestion or exposure to chemicals and industrial toxicants pose a serious health risk that may be hard to assess due to a lack of non-invasive diagnostic tests. Mapping chemical injuries to organ-specific damage and clinical outcomes via biomarkers or biomarker panels will provide the foundation for highly specific and robust diagnostic tests. Here, we have used DrugMatrix, a toxicogenomics database containing organ-specific gene expression data matched to dose-dependent chemical exposures and adverse clinical pathology assessments in Sprague Dawley rats, to identify groups of co-expressed genes (modules) specific to injury endpoints in the liver. We identified 78 such gene co-expression modules associated with 25 diverse injury endpoints categorized from clinical pathology, organ weight changes, and histopathology. Using gene expression data associated with an injury condition, we showed that these modules exhibited different patterns of activation characteristic of each injury. We further showed that specific module genes mapped to 1) known biochemical pathways associated with liver injuries and 2) clinically used diagnostic tests for liver fibrosis. As such, the gene modules have characteristics of both generalized and specific toxic response pathways. Using these results, we proposed three gene signature sets characteristic of liver fibrosis, steatosis, and general liver injury based on genes from the co-expression modules. Out of all 92 identified genes, 18 (20%) genes have well-documented relationships with liver disease, whereas the rest are novel and have not previously been associated with liver disease. In conclusion, identifying gene co-expression modules associated with chemically induced liver injuries aids in generating testable hypotheses and has the potential to identify putative biomarkers of adverse health effects. PMID:25226513

  2. Delivery of human mesenchymal adipose-derived stem cells restores multiple urological dysfunctions in a rat model mimicking radical prostatectomy damages through tissue-specific paracrine mechanisms

    DEFF Research Database (Denmark)

    Yiou, René; Mahrouf-Yorgov, Meriem; Trébeau, Céline;

    2016-01-01

    Urinary incontinence (UI) and erectile dysfunction (ED) are the most common functional urological disorders and the main sequels of radical prostatectomy (RP) for prostate cancer. Mesenchymal stem cell (MSC) therapy holds promise for repairing tissue damage due to RP. Because animal studies...... accurately replicating post-RP clinical UI and ED are lacking, little is known about the mechanisms underlying the urological benefits of MSC in this setting. To determine whether and by which mechanisms MSC can repair damages to both striated urethral sphincter (SUS) and penis in the same animal, we...

  3. Evaluation of engraftment of superparamagnetic iron oxide-labeled mesenchymal stem cells using three-dimensional reconstruction of magnetic resonance imaging in photo thrombotic cerebral infarction models of rats

    Energy Technology Data Exchange (ETDEWEB)

    Shim, Jae Hyun; Kwak, Byung Kook; Jung, Ji Sung; Park, Se Rah [Chung-Ang University College of Medicine, Seoul (Korea, Republic of)

    2015-06-15

    To evaluate engraftment by visualizing the location of human bone marrow-derived mesenchymal stem cells (hBM-MSCs) three-dimensionally in photothrombotic cerebral infarction (PTCI) models of rats. Magnetic resonance imaging (MRI) of an agarose block containing superparamagnetic iron oxide (SPIO)-labeled hBM-MSCs was performed using a 3.0-T MRI, T2-(T2WI), T2{sup *}-(T2{sup *}WI), and susceptibility-weighted images (SWI). PTCI was induced in 6 rats, and 2.5 x 10(5) SPIO-labeled hBM-MSCs were infused through the ipsilateral internal carotid artery (ICA group) or tail vein (IV group). MRI was performed on days 1, 3, 7, and 14 after stem cell injection. Dark signal regions were confirmed using histology. Three-dimensional MRI reconstruction was performed using the clinical workflow solution to evaluate the engraftment of hBM-MSCs. Volumetric analysis of the engraftment was also performed. The volumes of SPIO-labeled hBM-MSCs in the phantom MRI were 129.3, 68.4, and 25.9 microL using SWI, T2{sup *}WI, and T2WI, respectively. SPIO-labeled hBM-MSCs appeared on day 1 after injection, encircling the cerebral infarction from the ventral side. Dark signal regions matched iron positive cells and human origin (positive) cells. The volume of the engraftment was larger in the ICA group on days 1, 3, and 7, after stem cell injection (p < 0.05 on SWI). SWI was the most sensitive MRI pulse sequence (p < 0.05). The volume of infarction decreased until day 14. The engraftment of SPIO-labeled hBM-MSCs can be visualized and evaluated three-dimensionally in PTCI models of rats. The engraftment volume was larger in the ICA group than IV group on early stage within one week.

  4. Real-time-guided bone regeneration around standardized critical size calvarial defects using bone marrow-derived mesenchymal stem cells and collagen membrane with and without using tricalcium phosphate: an in vivo microcomputed tomographic and histologic experiment in rats

    Institute of Scientific and Technical Information of China (English)

    Khalid Al-Hezaimi; Sundar Ramalingam; Mansour Al-Askar; Aws S ArRejaie; Nasser Nooh; Fawad Jawad; Abdullah Aldahmash; Muhammad Atteya; Cun-Yu Wang

    2016-01-01

    The aimof the present real time in vivo micro-computed tomography (mCT) and histologic experiment was to assess the efficacy of guided bone regeneration (GBR) around standardized calvarial critical size defects (CSD) using bone marrow-derived mesenchymal stem cells (BMSCs), and collagen membrane (CM) with and without tricalcium phosphate (TCP) graft material. In the calvaria of nine female Sprague-Dawley rats, full-thickness CSD (diameter 4.6 mm) were created under general anesthesia. Treatment-wise, rats were divided into three groups. In group 1, CSD was covered with a resorbable CM; in group 2, BMSCs were filled in CSD and covered with CM; and in group 3, TCP soaked in BMSCs was placed in CSD and covered with CM. All defects were closed using resorbable sutures. Bone volume and bone mineral density of newly formed bone (NFB) and remaining TCP particles and rate of new bone formation was determined at baseline, 2, 4, 6, and 10 weeks using in vivo mCT. At the 10th week, the rats were killed and calvarial segments were assessed histologically. The results showed that the hardness ofNFBwas similar to that of the native bone in groups 1 and 2 as compared to theNFB in group 3. Likewise, values for the modulus of elasticity were also significantly higher in group 3 compared to groups 1 and 2. This suggests that TCP when used in combination with BMSCs and without CM was unable to form bone of significant strength that could possibly provide mechanical “lock” between the natural bone and NFB. The use of BMSCs as adjuncts to conventional GBR initiated new bone formation as early as 2 weeks of treatment compared to when GBR is attempted without adjunct BMSC therapy.

  5. Polyurethane/polylactide-based biomaterials combined with rat olfactory bulb-derived glial cells and adipose-derived mesenchymal stromal cells for neural regenerative medicine applications.

    Science.gov (United States)

    Grzesiak, Jakub; Marycz, Krzysztof; Szarek, Dariusz; Bednarz, Paulina; Laska, Jadwiga

    2015-01-01

    Research concerning the elaboration and application of biomaterial which may support the nerve tissue regeneration is currently one of the most promising directions. Biocompatible polymer devices are noteworthy group among the numerous types of potentially attractive biomaterials for regenerative medicine application. Polylactides and polyurethanes may be utilized for developing devices for supporting the nerve regeneration, like nerve guide conduits or bridges connecting the endings of broken nerve tracts. Moreover, the combination of these biomaterial devices with regenerative cell populations, like stem or precursor cells should significantly improve the final therapeutic effect. Therefore, the composition and structure of final device should support the proper adhesion and growth of cells destined for clinical application. In current research, the three polymer mats elaborated for connecting the broken nerve tracts, made from polylactide, polyurethane and their blend were evaluated both for physical properties and in vitro, using the olfactory-bulb glial cells and mesenchymal stem cells. The evaluation of Young's modulus, wettability and roughness of obtained materials showed the differences between analyzed samples. The analysis of cell adhesion, proliferation and morphology showed that the polyurethane-polylactide blend was the most neutral for cells in culture, while in the pure polymer samples there were significant alterations observed. Our results indicated that polyurethane-polylactide blend is an optimal composition for culturing and delivery of glial and mesenchymal stem cells.

  6. Intraperitoneal transplantation of human umbilical cord mesenchymal stem cells in the treatment of acute liver injury in rats%人脐带间充质干细胞腹腔移植治疗急性肝损伤大鼠

    Institute of Scientific and Technical Information of China (English)

    郑学峰; 刘霞

    2013-01-01

    BACKGROUND:In vitro experiments have confirmed that human umbilical cord mesenchymal stem cells can be induced to differentiate into hepatocyte-like cells, thus which can be considered to function as liver repair. OBJECTIVE:To observe the therapeutic effect of human umbilical cord mesenchymal stem celltransplantation on acute liver injuries in rats through in vivo animal experiments. METHODS:Healthy Sprague-Dawley rats were randomly divided into three groups:normal control group without modeling, celltransplantation group, and PBS group. Rat models of acute liver injury were prepared by 10%CCl4-olive oil solution in the celltransplantation and PBS groups which were fol owed by intraperitoneal injection of 0.5 mL human umbilical cord mesenchymal stem cellsuspension and 0.5 mL PBS, respectively. RESULTS AND CONCLUSION:Hematoxylin-eosin staining showed that pathological changes related to acute liver injury appeared at 24 hours after intraperitoneal injection of CCl 4 . Then, the liver structure recovered at 7 days after celltransplantation, but it did not recover til the 14th day after PBS injection. Compared with the normal control group, serum alanine aminotransferase and aspartate aminotransferase levels were significantly increased in the other two groups (P  目的:进行动物体内实验,观察人脐带间充质干细胞移植治疗大鼠急性肝损伤的效果。  方法:将健康SD大鼠随机分为3组,正常对照组不造模,细胞移植组和PBS组腹腔注射体积分数10%CCl4橄榄油溶液制造急性肝损伤模型后24 h,分别经腹腔移植人脐带间充质干细胞悬液0.5 mL和等量PBS。  结果与结论:苏木精-伊红染色显示,CCl4腹腔注射24 h后大鼠肝脏出现急性肝损伤的病理变化,细胞移植后7 d时肝脏结构完全恢复正常,PBS组14 d才恢复正常肝脏组织结构。与正常对照组相比,其他2组大鼠血清丙氨酸转氨酶及天门冬氨

  7. Isolation, cultivation, identify and cryopreservation of mesenchymal stem cells from rat adipose tissue in vitro%大鼠脂肪干细胞分离培养及细胞表型的研究

    Institute of Scientific and Technical Information of China (English)

    吕春燕; 陈高莉; 杨玲; 陈昌金; 袁慧; 杨雪梅

    2014-01-01

    Objective To search a method for isolation, cultivation, identification and cryopreservation of mesenchymal stem cells from rat adipose tissue in vitro. Method Three rats was anaesthetized and disinfected con-ventionally, their inguinal adipose tissue were taken and sheared to paste. The fragments were digested by collagenase typeⅠ, suspended by DMEM, and planted in culture dish. Then the cells were cultured in incubator culture. Their first medium change happen after 24~48 hours,then the change occur 2~3 days once again,after 5~7 days and 80%cell fu-sion, cells were digested by Trypsin and passaged by EDTA. Cell microscopy was used to morphological observation and the third generation was used to identify by flow cytometric cell cycle. Results Rats fat had a large amount of adipose-derived mesenchymal stem cells (ADMSCs). The shape of primary cultured ASCs was a star or much horn;af-ter passage of the ASCs relatively uniform shape,fusiform shape,spiral growth. The ASCs surface markers are identi-fied by flow cytometry and showed positive expression of CD44, CD106. Cryopreserved ASCs remains a good activity, high survival rate. Conclusion A simple and convenient method was successfully established to isolate, culture and store the adipose tissue derived mesenchymal stem cells, which provides the foundation for the future use of ADMSCs in the further research.%目的:分离、培养和冻存大鼠脂肪干细胞,并对其相关的细胞表型进行研究,为利用脂肪干细胞治疗梗阻性肾病肾纤维化等实验提供依据。方法取大鼠3只,经消毒麻醉,采集其腹股沟处脂肪组织分离培养其中的脂肪干细胞,用相差倒置显微镜观察细胞生长方式及形态变化。取第三代细胞用流式细胞仪作细胞免疫表型的鉴定。冻存复苏后再次检测干细胞生长情况及表型。结果大鼠脂肪组织中含有大量间充质干细胞,原代培养的ASCs呈小圆形或星形,经传代后的脂肪

  8. Bone marrow mesenchymal stem cell transplantation for gastric precancerous lesions in a rat model%骨髓间充质干细胞移植对大鼠胃癌前病变模型的修复作用

    Institute of Scientific and Technical Information of China (English)

    刘洪锋; 李潞

    2016-01-01

    背景:胃癌前病变是多种因素共同参与的长期发展过程,骨髓间充质干细胞有修复组织损伤的作用。目的:探讨骨髓间充质干细胞移植对大鼠胃癌前病变的影响。  方法:将36只Wistar大鼠随机分为对照组、模型组和移植组。模型组和移植组建立大鼠胃癌前病变模型,移植组大鼠经尾静脉注射1 mL CM-dil标记的骨髓间充质干细胞(3×106个细胞),每周注射1次,一共3次,模型组和对照组同样方法经尾静脉注入等量的生理盐水。末次注射后1周处死大鼠,观察大鼠胃组织病理变化,检测胃黏膜中血管内皮生长因子的表达以及血清中细胞因子含量。  结果与结论:①移植组大鼠胃黏膜损伤较模型组轻,移植组血管内皮生长因子表达明显高于模型组和对照组(P<0.05);②模型组和移植组血清白细胞介素17和γ-干扰素水平高于对照组(P<0.05),移植组血清白细胞介素17和γ-干扰素水平低于模型组(P<0.05);③结果表明,骨髓间充质干细胞通过增加胃黏膜病变部位血管内皮生长因子的表达,降低血清白细胞介素17和γ-干扰素水平,从而提高血管通透性,减轻炎症反应,阻断或者缓解癌前病变的发生。%BACKGROUND:Precancerous lesions are a long-term development process in which many factors are involved. Bone marrow mesenchymal stem cel s can repair tissue injury. OBJECTIVE:To investigate the effect of bone marrow mesenchymal stem cel transplantation on gastric precancerous lesions in the rats. METHODS:Thirty-six Wistar rats were randomly divided into control group, model group and transplantation group. Animal models of gastric precancerous lesions were established in the model and transplantation groups. Rats in the transplantation group were given 1 mL of CM-dil-labeled bone marrow mesenchymal stem cel s (3×106 cel s) via the tail vein, once a week, total y three times. Rats in the

  9. 骨髓间充质干细胞移植大鼠脑梗死区的超微结构变化%Changes of ultrastructure in cerebral infarction region of rats with transplantation mesenchymal stem cells

    Institute of Scientific and Technical Information of China (English)

    桂莉; 范文辉; 李露斯; 陈康宁

    2004-01-01

    背景:近来的研究发现骨髓间充质干细胞(mesenchymal stem cells,MSCs)对缺血性脑血管病有治疗作用,接受细胞移植的脑梗死区超微结构的观察可进一步证实该作用.目的:观察接受MSCs局部注射移植的大鼠脑梗死区超微结构的变化,以了解MSCs移植对梗死区超微结构的影响.设计:随机对照实验研究.单位:解放军第三军医大学西南医院神经病学中心.材料:实验于2002-04/2002-09在解放军第三军医大学动物所及中心实验室完成.实验动物为健康幼年及成年Wistar大鼠12只,雌雄不限.随机分为移植组6只和对照组6只.干预:建立大鼠大脑中动脉栓塞模型,将体外分离培养的大鼠MSCs注射至大鼠脑梗死区,两组大鼠于移植后第4周电镜观察大鼠脑梗死区超微结构变化.主要观察指标:移植组及对照组大鼠脑梗死区超微结构变化.结果:接受移植的梗死区可见外形幼稚的细胞存活,并围绕在神经元周围,梗死区神经元胞浆中出现了丰富的游离核糖体,而对照组大鼠梗死区神经元胞核固缩,并有嗜神经细胞现象.结论:大鼠MSCs移植对缺血性脑损伤具有一定修复作用.%BACKGROUND: It is recently found that mesenchymal stem cells(MSCs)have therapeutic function for ischemic cerebrovascular disease and that the function should have a further observation of the ultrastructural changes in cerebral infarction region with quantitative MSCs transplantation.OBJECTIVE: To observe the ultrastructural changes in cerebral infarction region of rats injected locally with mesenchymal stem cells so as to understand the effect of MSCs transplantation on the ultrastructure in cerebral infarction region.DESIGN: A randomized and controlled trial.SETTING: Neurology Center of Southwest Hospital Affiliated to the Third Military Medical University of Chinese PLA.MATERIALS: The experiment was done in the medical Animal Institute and Central Laboratory of the Third

  10. Uterine mesenchymal tumors

    Directory of Open Access Journals (Sweden)

    Nikhil A Sangle

    2011-01-01

    Full Text Available Uterine mesenchymal tumors are a heterogeneous group of neoplasms that can frequently be diagnostically challenging. Differentiation between the benign and malignant counterparts of mesenchymal tumors is significant due to differences in clinical outcome, and the role of the surgical pathologist in making this distinction (especially in the difficult cases cannot be underestimated. Although immunohistochemical stains are supportive toward establishing a final diagnosis, the morphologic features trump all the other ancillary techniques for this group of neoplasms. This review therefore emphasizes the key morphologic features required to diagnose and distinguish uterine mesenchymal tumors from their mimics, with a brief description of the relevant immunohistochemical features.

  11. Bone marrow mesenchymal stem cells cultured in artificial meninges repair infarcted myocardium in rats%人工脑膜复合大鼠骨髓间充质干细胞修复心肌梗死★

    Institute of Scientific and Technical Information of China (English)

    马红芬; 张晓刚; 史若飞; 熊挺淋; 赵霞

    2013-01-01

      背景:干细胞移植治疗心肌梗死拥有广泛的应用前景,寻求理想的细胞类型和有效的移植方式是提高干细胞治疗效果的关键因素。目的:探讨人工脑膜复合骨髓间充质干细胞修复心肌梗死的安全性及作用。方法:采用全骨髓贴壁筛选法分离培养骨髓间充质干细胞,取培养良好的第3代骨髓间充质干细胞经DAPI 标记后接种于人工脑膜制备细胞人工脑膜复合物。构建 SD 大鼠心肌梗死模型,60只大鼠随机数字表法均分为假手术组、心肌梗死组、人工脑膜组、细胞脑膜复合物组。移植4周后检测心功能参数, Western blot 检测心肌组织缝隙连接蛋白43的表达,计算心肌梗死后生存率。结果与结论:构建心肌梗死模型并移植后4周,细胞脑膜复合物组心脏组织冰冻切片于荧光显微镜下可观察到心肌内少量核蓝染的细胞,表明骨髓间充质干细胞得以存活;细胞脑膜复合物组与心肌梗死组和人工脑膜组相比,左心室功能明显改善,Cx43蛋白的表达上调,生存率增加(P <0.05)。说明人工脑膜复合骨髓间充质干细胞移植可提高心肌梗死大鼠心脏功能及生存率。%BACKGROUND: Stem cel transplantation has broad application prospects in the treatment of myocardial infarction. To find the ideal cel type and the effective transplantation method is the key factor to improve. OBJECTIVE: To investigate the effect and safety of artificial meninges combined with bone marrow mesenchymal stem cel s on repairing infarcted heart. METHODS: Bone marrow mesenchymal stem cel s were obtained by whole bone marrow culture method, and subcultured to the third generation. Then bone marrow mesenchymal stem cel s were labeled with 2-(4-amidinophenyl)-6-indolecarbamidine dihydrochloride and seeded on the cel s and artificial meningeal complex. Sprague Dawley rats model of myocardial infarction were established, and 60 rats

  12. An experimental study of preventing and treating acute radioactive enteritis with human umbilical cord mesenchymal stem cells

    Institute of Scientific and Technical Information of China (English)

    Rui Wang; Wei Yuan; Qiang Zhao; Peng Song; Ji Yue; Shi-De Lin; Ting-Bao Zhao

    2013-01-01

    Objective:To test the curative effect of human umbilical cord-derived mesenchymal stem cells on rat acute radioactive enteritis and thus to provide clinical therapeutic basis for radiation sickness.Methods:Human umbilical cord-derived mesenchymal stem cells were cultivatedin vitro and the model of acute radioactive enteritis of rats was established.Then, the umbilical cord mesenchymal stem cells were injected into the rats via tail vein.Visual and histopathological changes of the experimental rats were observed.Results:After the injection, the rats in the prevention group and treatment group had remarkably better survival status than those in the control group.The histological observations revealed that the former also had better intestinal mucosa structure, more regenerative cells and stronger proliferation activity than the latter.Conclusions:Human umbilical cord-derived mesenchymal stem cells have a definite therapeutic effect on acute radioactive enteritis in rats.

  13. Conditioned medium from mesenchymal stem cells induces cell death in organotypic cultures of rat hippocampus and aggravates lesion in a model of oxygen and glucose deprivation.

    Science.gov (United States)

    Horn, Ana Paula; Frozza, Rudimar Luiz; Grudzinski, Patrícia Benke; Gerhardt, Daniéli; Hoppe, Juliana Bender; Bruno, Alessandra Nejar; Chagastelles, Pedro; Nardi, Nance Beyer; Lenz, Guido; Salbego, Christianne

    2009-01-01

    Cell therapy using bone marrow-derived mesenchymal stem cells (MSC) seems to be a new alternative for the treatment of neurological diseases, including stroke. In order to investigate the response of hippocampal tissue to factors secreted by MSC and if these factors are neuroprotective in a model of oxygen and glucose deprivation (OGD), we used organotypic hippocampal cultures exposed to conditioned medium from bone marrow-derived MSC. Our results suggest that the conditioned medium obtained from these cells aggravates lesion caused by OGD. In addition, the presence of the conditioned medium alone was toxic mainly to cells in the CA1, CA2 and CA3 areas of the hippocampal organotypic culture even in basal conditions. GABA stimulation and NMDA and AMPA receptors antagonists were able to reduce propidium iodide staining, suggesting that the cell death induced by the toxic factors secreted by MSC could involve these receptors.

  14. Apobec-1 Complementation Factor (A1CF Inhibits Epithelial-Mesenchymal Transition and Migration of Normal Rat Kidney Proximal Tubular Epithelial Cells

    Directory of Open Access Journals (Sweden)

    Liyuan Huang

    2016-02-01

    Full Text Available Apobec-1 complementation factor (A1CF is a member of the heterogeneous nuclear ribonucleoproteins (hnRNP family, which participates in site-specific posttranscriptional RNA editing of apolipoprotein B (apoB transcript. The posttranscriptional editing of apoB mRNA by A1CF in the small intestine is required for lipid absorption. Apart from the intestine, A1CF mRNA is also reported to be highly expressed in the kidneys. However, it is remained unknown about the functions of A1CF in the kidneys. The aim of this paper is to explore the potential functions of A1CF in the kidneys. Our results demonstrated that in C57BL/6 mice A1CF was weakly expressed in embryonic kidneys from E15.5dpc while strongly expressed in mature kidneys after birth, and it mainly existed in the tubules of inner cortex. More importantly, we identified A1CF negatively regulated the process of epithelial-mesenchymal transition (EMT in kidney tubular epithelial cells. Our results found ectopic expression of A1CF up-regulated the epithelial markers E-cadherin, and down-regulated the mesenchymal markers vimentin and α-smooth muscle actin (α-SMA in NRK52e cells. In addition, knockdown of A1CF enhanced EMT contrary to the overexpression effect. Notably, the two A1CF variants led to the similar trend in the EMT process. Taken together, these data suggest that A1CF may be an antagonistic factor to the EMT process of kidney tubular epithelial cells.

  15. 脐带间充质干细胞移植可延缓大鼠失神经肌肉的萎缩%Human umbilical cord mesenchymal stem cells transplantation delays denervated muscle atrophy in rats

    Institute of Scientific and Technical Information of China (English)

    陈传煌; 杨万章; 杨涛; 吴芳; 李文庆; 李楚炎; 毛仁群; 余志才; 张国雷; 肖振兴

    2014-01-01

    背景:周围神经断伤后生长缓慢,失神经支配的肌肉萎缩及运动终板纤维化,导致肢体功能不可逆障碍。脐带间充质干细胞已经广泛应用于多学科研究,但应用于周围神经损伤中延缓大鼠失神经肌肉萎缩鲜有报道。目的:观察异种异基因脐带间充干细胞移植于大鼠离断坐骨神经断端,延缓失神经肌肉萎缩的效果。  方法:新鲜脐带采集于健康足月产妇,分离鉴定脐带间充质干细胞。制备大鼠坐骨神经SunderlandⅣ度损伤模型,去神经束5 mm,神经外膜修复,5 mm小间隙移植脐带间充质干细胞模型,对照组仅在小间隙内注入同体积生理盐水。测定大鼠坐骨神经功能指数,小腿三头肌湿质量,坐骨神经干潜伏期、动作电位传导速度、波幅,以及骨骼肌纤维横截面积维持率。  结果与结论:造模后4,8及12周,脐带间充质干细胞组大鼠坐骨神经功能指数、右侧小腿三头肌湿质量及骨骼肌纤维横截面积维持率均显著高于对照组(P OBJECTIVE:To observe the value of human umbilical cord mesenchymal stem cells transplantation to delay denervated muscle atrophy of rats after sciatic nerve injury. METHODS:Umbilical cord blood was col ected from healthy parturient woman after ful-term delivery. In the experimental group, the rat’s Sunderland IV degree sciatic nerve injury model was established by 5 mm denervation, epineurial repair, and 5 mm smal gap transplantation of umbilical cord mesenchymal stem cells. In the control group, after modeling, the same volume of normal saline was injected into the smal gap. The main outcome measures included the sciatic nerve function index, the wet weight of triceps surae,sciatic nerve latency, action potential conduction velocity and amplitude,and skeletal muscle fiber cross section area maintenance rate. RESULTS AND CONCLUSION:After 4, 8 and 12 weeks of modeling, the sciatic nerve

  16. The anti-oxidative role of micro-vesicles derived from human Wharton-Jelly mesenchymal stromal cells through NOX2/gp91(phox suppression in alleviating renal ischemia-reperfusion injury in rats.

    Directory of Open Access Journals (Sweden)

    Guangyuan Zhang

    Full Text Available Oxidative stress is known as one of the main contributors in renal ischemia/reperfusion injury (IRI. Here we hypothesized that Micro-vesicles (MVs derived from human Wharton Jelly mesenchymal stromal cells (hWJMSCs could protect kidney against IRI through mitigating oxidative stress. MVs isolated from hWJMSCs conditioned medium were injected intravenously in rats immediately after unilateral kidney ischemia for 60 min. The animals were sacrificed at 24 h, 48 h and 2 weeks respectively after reperfusion. Our results show that the expression of NOX2 and reactive oxygen species (ROS in injured kidney tissues was declined and the oxidative stress was alleviated in MVs group at 24 h and 48 h in parallel with the reduced apoptosis and enhanced proliferation of cells. IRI-initiated fibrosis was abrogated by MVs coincident with renal function amelioration at 2 weeks. NOX2 was also found down-regulated by MVs both in human umbilical vein endothelial cells (HUVEC and NRK-52E cell line under hypoxia injury model in vitro. In conclusion, a single administration of hWJMSC-MVs might protect the kidney by alleviation of the oxidative stress in the early stage of kidney IRI through suppressing NOX2 expression. Moreover, it could reduce the fibrosis and improved renal function.

  17. Allelic Variants in Arhgef11 via the Rho-Rock Pathway Are Linked to Epithelial-Mesenchymal Transition and Contributes to Kidney Injury in the Dahl Salt-Sensitive Rat.

    Directory of Open Access Journals (Sweden)

    Zhen Jia

    Full Text Available Previously, genetic analyses identified that variants in Arhgef11 may influence kidney injury in the Dahl salt-sensitive (S rat, a model of hypertensive chronic kidney disease. To understand the potential mechanism by which altered expression and/or protein differences in Arhgef11 could play a role in kidney injury, stably transduced Arhgef11 knockdown cell lines as well as primary cultures of proximal tubule cells were studied. Genetic knockdown of Arhgef11 in HEK293 and NRK resulted in reduced RhoA activity, decreased activation of Rho-ROCK pathway, and less stress fiber formation versus control, similar to what was observed by pharmacological inhibition (fasudil. Primary proximal tubule cells (PTC cultured from the S exhibited increased expression of Arhgef11, increased RhoA activity, and up regulation of Rho-ROCK signaling compared to control (small congenic. The cells were also more prone (versus control to TGFβ-1 induced epithelial-mesenchymal transition (EMT, a hallmark feature of the development of renal interstitial fibrosis, and characterized by development of spindle shape morphology, gene expression changes in EMT markers (Col1a3, Mmp9, Bmp7, and Ocln and increased expression of N-Cadherin and Vimentin. S derived PTC demonstrated a decreased ability to uptake FITC-albumin compared to the small congenic in vitro, which was confirmed by assessment of albumin re-uptake in vivo by infusion of FITC-albumin and immunofluorescence imaging. In summary, these studies suggest that genetic variants in the S form of Arhgef11 via increased expression and/or protein activity play a role in promoting kidney injury in the S rat through changes in cell morphology (Rho-Rock and/or EMT that impact the function of tubule cells.

  18. TNF-α刺激大鼠骨髓间充质干细胞的机制研究%Mechanism of TNF-α in Stimulation of Bone Marrow Mesenchymal Stem Cells in Rats

    Institute of Scientific and Technical Information of China (English)

    彭程飞; 韩雅玲; 邓捷; 闫承慧; 康建; 李洋; 李杰; 王艳春

    2011-01-01

    Objective: To investigate the promotlve effects of NF-κ B on the marrow-derived mesenchymal stem cells.Methods:Mononuclear cells were isolated from rat bone marrow by density-gradient centrifugation and were cultured on fibronectin-coated plates,supplied with bovine pituitary extract.MSCs were identified by Morphology and Surface molecule markers of MSCs.After the marrow-derived mesenchymal stem cells were stimulated by TNF-α , the production of the growth factors and the protein expression of NF-κ B in different groups were compared by ELISA and WESTERN BOLT.Result: ① The marrow-derived mesenchymal stem cells were successful cultured by observing the morphology of the marrow-derived mesenchymal stem cells and detecting the cell surface antigen by flow cytometry assays.② The production of the growth factors had significantly increased in TNF-α stimulation group compared with that of no TNF-α stimulation group.The expression ofp-Iκ B increased significantly indicating the activation of NF-κ B (P<0.05).The production of the growth factors had significantly decreased in stimulation of TNF-α plus the inhibitor of NF-κ B group compared stimulation of TNF-α group,.And the protein expression of p-Iκ B significantly decressed indicate the inhibition of NF-κ B (P<0.05).Conclusion: NF-κ B had crucially protective effects on the marrow-derived mesenchymal stem cells stimulated by TNF-α.%目的:研究肿瘤坏死因子-α(Tumor necrosis fagtorα,TNFα)刺激大鼠骨髓间充质干细胞(marrow-derived mesenchymal stem cells,MSCs)的作用机制.方法:采取大鼠骨髓,以密度梯度离心分离出单个核细胞(MNCs),于体外培养并由牛垂体提取物(PEX)诱导扩增传代培养出骨髓间充质干细胞(MSCs).经形态学和流式细胞仪检测MSCs表面标志物鉴定后,用TNF-α刺激骨髓间充质干细胞(MSCs),通过酶联免疫吸附剂测定法(enzyme linked immunosorbent,assay.ELISA)观察比较不同组别细胞的生长因

  19. Epiretinal transplantation of human bone marrow mesenchymal stem cells rescues retinal and vision function in a rat model of retinal degeneration

    Directory of Open Access Journals (Sweden)

    Adi Tzameret

    2015-09-01

    Our findings suggest that transplantation of hBM-MSCs as a thin epiretinal layer is effective for treatment of retinal degeneration in RCS rats, and that transplanting the cells in close proximity to the retina enhances hBM-MSC therapeutic effect compared with intravitreal injection.

  20. Effect of Single and Double Administration of Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells Following Focal Cerebral Ischemia in Rats

    Science.gov (United States)

    Park, Hyung Woo; Kim, Yona; Chang, Jong Wook; Yang, Yoon Sun; Oh, Wonil; Lee, Jae Min; Park, Hye Ran; Kim, Dong Gyu

    2017-01-01

    Stem cell therapies are administered during the acute phase of stroke to preserve the penumbral tissues from ischemic injury. However, the effect of repeated cell therapy during the acute phase remains unclear. In this study, we investigated and compared the functional outcome of single (two days post-injury) and repeated (two and nine days post-injury) treatment with human umbilical cord derived mesenchymal stem cells (hUCB-MSCs) after middle cerebral artery occlusion (MCAO). The rotarod and limb placement tests were utilized to investigate functional outcomes, while infarct volume and tissue damage were measured by immunofluorescent staining for neovascularization, neurogenesis, apoptosis, and inflammation in the penumbral zones. We observed notable motor dysfunction and a significant decrease in infarcted brain volume, as well as increases in neurons and vessels in both single and repeated hUCB-MSC treatments compared to the control group. Interestingly, repeated administration of hUCB-MSCs was not found to elicit additional or synergistic improvements over monotherapy. This study suggests that a clearer understanding of the therapeutic window after stroke will facilitate the development of more efficient treatment protocols in the clinical application of stem cell therapy. PMID:28243167

  1. Effects of mesenchymal stem cell therapy, in association with pharmacologically active microcarriers releasing VEGF, in an ischaemic stroke model in the rat.

    Science.gov (United States)

    Quittet, Marie-Sophie; Touzani, Omar; Sindji, Laurence; Cayon, Jérôme; Fillesoye, Fabien; Toutain, Jérôme; Divoux, Didier; Marteau, Léna; Lecocq, Myriam; Roussel, Simon; Montero-Menei, Claudia N; Bernaudin, Myriam

    2015-03-01

    Few effective therapeutic interventions are available to limit brain damage and functional deficits after ischaemic stroke. Within this context, mesenchymal stem cell (MSC) therapy carries minimal risks while remaining efficacious through the secretion of trophic, protective, neurogenic and angiogenic factors. The limited survival rate of MSCs restricts their beneficial effects. The usefulness of a three-dimensional support, such as a pharmacologically active microcarrier (PAM), on the survival of MSCs during hypoxia has been shown in vitro, especially when the PAMs were loaded with vascular endothelial growth factor (VEGF). In the present study, the effect of MSCs attached to laminin-PAMs (LM-PAMs), releasing VEGF or not, was evaluated in vivo in a model of transient stroke. The parameters assessed were infarct volume, functional recovery and endogenous cellular reactions. LM-PAMs induced the expression of neuronal markers by MSCs both in vitro and in vivo. Moreover, the prolonged release of VEGF increased angiogenesis around the site of implantation of the LM-PAMs and facilitated the migration of immature neurons towards the ischaemic tissue. Nonetheless, MSCs/LM-PAMs-VEGF failed to improve sensorimotor functions. The use of LM-PAMs to convey MSCs and to deliver growth factors could be an effective strategy to repair the brain damage caused by a stroke.

  2. Chemical -induced apoptotic cell death in tomato cells : involvement of caspase-like proteases

    NARCIS (Netherlands)

    Jong, de A.J.; Hoeberichts, F.A.; Yakimova, E.T.; Maximova, E.; Woltering, E.J.

    2000-01-01

    A new system to study programmed cell death in plants is described. Tomato (Lycopersicon esculentum Mill.) suspension cells were induced to undergo programmed cell death by treatment with known inducers of apoptosis in mammalian cells. This chemical-induced cell death was accompanied by the characte

  3. Chemical-Induced Erythrocytosis in Wistar Rats: Assessment as a Model for Human Polycythemia.

    Science.gov (United States)

    1985-05-01

    0.5 ppm have reduced the potential of acquiring Minamata disease [135]. Accidental poisonings from consumption of organomercurial-treated seed grains...tubules [111]. Neuropathology. Histopathology reveals demyelination and vacuolization of dorsal nerve roots and peripheral nerves [111], cerebrovascular

  4. Profiles of metabolites and gene expression in rats with chemically induced hepatic necrosis

    NARCIS (Netherlands)

    Heijne, W.H.M.; Lamers, R.J.A.N.; Bladeren, P.J. van; Groten, J.P.; Nesselrooij, J.H.J. van; Ommen, B. van

    2005-01-01

    This study investigated whether integrated analysis of transcriptomics and metabolomics data increased the sensitivity of detection and provided new insight in the mechanisms of hepatotoxicity. Metabolite levels in plasma or urine were analyzed in relation to changes in hepatic gene expression in ra

  5. Distribution of transplanted human mesenchymal stem cells from Wharton’s Jelly in the central nervous systems of the EAE rats

    Directory of Open Access Journals (Sweden)

    Kovalchuk M. V.

    2015-10-01

    Full Text Available Human Wharton’s Jelly MSCs (hWJ-MSCs have a considerable advantage and potential in treating the central nervous system diseases and can be a new alternative treatment of Multiple Sclerosis (MS. Aim. To study the persistence and distribution of hWJ-MSCs along the neuraxis following transplantation in central nervous system of rats with experimental autoimmune encephalomyelitis (EAE, the animal model of MS. Methods. Isolation and cultivation of hWJ-MSCs in vitro. Immunological phenotyping by flow cytometry. EAE induction. Intrathecal (suboccipital injection of MSCs into CNS of SCH-induced EAE rats. Persistence of hWJ-MSCs in the CNS of hWJ-MSCs -treated rats was detected through detection of the human alpha-satellite DNA in the tissue sections and the cerebrospinal fluid (CSF by PCR at days 2, 3, 4 and 5 Results. PCR-assays for alpha-satellite sequences revealed that Human DNA was detected during 5 days following intrathecal injection at the peak of disease in the treated rats. It has been demonstrated that the human DNA was traced in CSF and various segments of a spinal cord. Conclusions. The data obtained suggest that intrathecally delivered hWJ-MSCs, with time, can migrate through the CSF from the injection site to various segments of CNS and persist therein during the first week of post transplantation, which was performed at the EAE disease peak in the xenogeneic setting without immunosuppression. hWJ-MSCs may be considered as a delivery cell source of therapeutic molecules for CNS inflammatory diseases.

  6. CD-REST: a system for extracting chemical-induced disease relation in literature

    Science.gov (United States)

    Xu, Jun; Wu, Yonghui; Zhang, Yaoyun; Wang, Jingqi; Lee, Hee-Jin; Xu, Hua

    2016-01-01

    Mining chemical-induced disease relations embedded in the vast biomedical literature could facilitate a wide range of computational biomedical applications, such as pharmacovigilance. The BioCreative V organized a Chemical Disease Relation (CDR) Track regarding chemical-induced disease relation extraction from biomedical literature in 2015. We participated in all subtasks of this challenge. In this article, we present our participation system Chemical Disease Relation Extraction SysTem (CD-REST), an end-to-end system for extracting chemical-induced disease relations in biomedical literature. CD-REST consists of two main components: (1) a chemical and disease named entity recognition and normalization module, which employs the Conditional Random Fields algorithm for entity recognition and a Vector Space Model-based approach for normalization; and (2) a relation extraction module that classifies both sentence-level and document-level candidate drug–disease pairs by support vector machines. Our system achieved the best performance on the chemical-induced disease relation extraction subtask in the BioCreative V CDR Track, demonstrating the effectiveness of our proposed machine learning-based approaches for automatic extraction of chemical-induced disease relations in biomedical literature. The CD-REST system provides web services using HTTP POST request. The web services can be accessed from http://clinicalnlptool.com/cdr. The online CD-REST demonstration system is available at http://clinicalnlptool.com/cdr/cdr.html. Database URL: http://clinicalnlptool.com/cdr; http://clinicalnlptool.com/cdr/cdr.html PMID:27016700

  7. Epiretinal transplantation of human bone marrow mesenchymal stem cells rescues retinal and vision function in a rat model of retinal degeneration.

    Science.gov (United States)

    Tzameret, Adi; Sher, Ifat; Belkin, Michael; Treves, Avraham J; Meir, Amilia; Nagler, Arnon; Levkovitch-Verbin, Hani; Rotenstreich, Ygal; Solomon, Arieh S

    2015-09-01

    Vision incapacitation and blindness associated with incurable retinal degeneration affect millions of people worldwide. In this study, 0.25×10(6) human bone marrow stem cells (hBM-MSCs) were transplanted epiretinally in the right eye of Royal College Surgeons (RCS) rats at the age of 28 days. Epiretinally transplanted cells were identified as a thin layer of cells along vitreous cavity, in close proximity to the retina or attached to the lens capsule, up to 6 weeks following transplantation. Epiretinal transplantation delayed photoreceptor degeneration and rescued retinal function up to 20 weeks following cell transplantation. Visual functions remained close to normal levels in epiretinal transplantation rats. No inflammation or any other adverse effects were observed in transplanted eyes. Our findings suggest that transplantation of hBM-MSCs as a thin epiretinal layer is effective for treatment of retinal degeneration in RCS rats, and that transplanting the cells in close proximity to the retina enhances hBM-MSC therapeutic effect compared with intravitreal injection.

  8. Dose-dependent Effects of Strontium Ranelate on Ovariectomy Rat Bone Marrow Mesenchymal Stem Cells and Human Umbilical Vein Endothelial Cells

    Science.gov (United States)

    Guo, Xiaojing; Wei, Silong; Lu, Mengmeng; Shao, Zhengwei; Lu, Jiayu; Xia, Lunguo; Lin, Kaili; Zou, Derong

    2016-01-01

    In clinic, strontium ranelate (SrR) is a useful drug to treat osteoporosis by orally taken method, but some side effect appeared in recent years. The aim of this study is to evaluate the effectiveness and safety of SrR on cells by direct application, to study the possibility of local application of this drug. Qualitative ALP staining, quantitative ALP activity assay, alizarin red staining, realtime PCR and westernblot assay were used to evaluate the osteogenesis ability of SrR under normal or osteogenic induction environment of ovariectomy bone marrow mesenchymal stem cells (OVX-BMSCs). The angiogenesis ability of SrR was studied by immunofluorescence staining of CD31 and vWF of OVX-BMSCs under angiogenesis induction environment, transwell, tubeformation and realtime PCR assay of HUVECs. Signaling pathway of PI3K/AKT/mTOR was also studied. The result demonstrated that SrR could enhance proliferation and osteogenic differentiation of OVX-BMSCs. The osteogenesis effect of SrR has been proved by the better performed of ALP activity, alizarin red staining and the remarkable up-regulation of ALP, Col-I, Runx2, OCN, BMP-2, BSP, OPG of the OVX-BMSCs, and reduction of RANKL. In addition, SrR promotes angiogenesis differentiation of both OVX-BMSCs and HUVECs. Higher intensity of immunostaining of CD31 and vWF, better result of transwell and tubeformation assay could be observed in SrR treated group, and increasing mRNA levels of VEGF and Ang-1 in the OVX-BMSCs, VEGF in HUVECs were learnt. Signaling pathway assay showed that PI3K/AKT/mTOR signaling pathway was involved in this SrR triggered angiogenesis procedure. The thrombosis marker ET-1, PAI-1 and t-PA were up-regulated, but no significant differences for low concentration (regeneration. PMID:27994515

  9. Bone marrow-derived mesenchymal stem cells repaired but did not prevent gentamicin-induced acute kidney injury through paracrine effects in rats.

    Directory of Open Access Journals (Sweden)

    Luciana A Reis

    Full Text Available This study evaluated the effects of bone marrow-derived mesenchymal stem cells (BMSCs or their conditioned medium (CM on the repair and prevention of Acute Kidney Injury (AKI induced by gentamicin (G. Animals received daily injections of G up to 20 days. On the 10(th day, injections of BMSCs, CM, CM+trypsin, CM+RNase or exosome-like microvesicles extracted from the CM were administered. In the prevention groups, the animals received the BMSCs 24 h before or on the 5(th day of G treatment. Creatinine (Cr, urea (U, FENa and cytokines were quantified. The kidneys were evaluated using hematoxylin/eosin staining and immunohystochemistry. The levels of Cr, U and FENa increased during all the periods of G treatment. The BMSC transplantation, its CM or exosome injections inhibited the increase in Cr, U, FENa, necrosis, apoptosis and also increased cell proliferation. The pro-inflammatory cytokines decreased while the anti-inflammatory cytokines increased compared to G. When the CM or its exosomes were incubated with RNase (but not trypsin, these effects were blunted. The Y chromosome was not observed in the 24-h prevention group, but it persisted in the kidney for all of the periods analyzed, suggesting that the injury is necessary for the docking and maintenance of BMSCs in the kidney. In conclusion, the BMSCs and CM minimized the G-induced renal damage through paracrine effects, most likely through the RNA carried by the exosome-like microvesicles. The use of the CM from BMSCs can be a potential therapeutic tool for this type of nephrotoxicity, allowing for the avoidance of cell transplantations.

  10. Bone marrow mesenchymal stem cells from diabetic versus normal rats for treatment of myocardial infarction%糖尿病与正常大鼠骨髓间充质干细胞移植治疗心肌梗死的比较★

    Institute of Scientific and Technical Information of China (English)

    杨忠路; 王辉山

    2013-01-01

    BACKGROUND: Current studies have shown that bone marrow mesenchymal stem cells from normal or young people usual y serve as a source of transplanted cells in stem cel transplantation treatment of myocardial infarction. OBJECTIVE: To compare the therapeutic effects of bone marrow mesenchymal stem cells from diabetic and normal rats on myocardial infarction. METHODS: Under sterile conditions, bone marrow mesenchymal stem cells from normal and diabetic rats were harvested. Then, rat models of myocardial infarction were established and randomly divided into three groups:100 μL cellsuspension containing 105-106 bone marrow mesenchymal stem cells from F2 normal or diabetic rats was injected into myocardial infarction lesions, and 100 μL Dulbecco’s modified Eagle’s medium containing 20%fetal bovine serum was injected serving as blank control. After 1 month, hematoxylin-eosin staining for myocardial infarction lesions was performed for histomorphological observation. Bcl-2 expression was detected by immunohistochemistry method. RESULTS AND CONCLUSION: Based on cel morphology observation and flow cytometry identification, high-purity bone marrow mesenchymal stem cells could be obtained using rat femoral bone marrow adherent culture. Cel growth curve showed that normal rat bone marrow mesenchymal stem cells grew faster than those from diabetic rats. At 1 month after transplantation, histomorphological improvement was seen in the infarcted area after transplantation of normal rat bone marrow mesenchymal stem cells as compared with the other two groups. In addition, the Bcl-2 expression in the infarcted area was higher in the normal rat cel group than the the other two groups. These findings indicate that bone marrow mesenchymal stem cells from normal rats grow faster than those from diabetic rats, and the cells from normal rats have better therapeutic effects on myocardial infarction.%  背景:目前国内外在干细胞移植治疗心肌梗死的研究中,大

  11. 骨髓间充质干细胞对肾病综合征大鼠CD4+CD25+调节性T细胞的影响%Effects of bone marrow mesenchymal stem cell transplantation on CD4+CD25+regulatory T cells in rats with primary nephrotic syndrome

    Institute of Scientific and Technical Information of China (English)

    杨焕丹; 张锐锋; 封东进; 朱冰冰; 吕娟

    2014-01-01

    BACKGROUND:Decreased function and reduced number of CD4+CD25+regulatory T cells have been considered the major manifestation of immunity dysfunction in children with primary nephrotic syndrome. Bone marrow mesenchymal stem cells have immunoregulation effects, which up-regulate CD4+CD25+regulatory T cells, inhibit proliferation of lymphocytes, and have been widely used in many immune diseases. OBJECTIVE:To investigate the effects of bone marrow mesenchymal stem celltransplantation on the CD4+CD25+regulatory T cells of peripheral blood in rats with primary nephrotic syndrome. METHODS:Bone marrow mesenchymal stem cells from six Sprague-Dawley rats were isolated, passaged and utilized for cellsuspension preparation. At the third passage, bone marrow mesenchymal stem cells were used for transplantation. The remaining 30 rats were randomly and equal y divided into three groups:normal group, normal saline infusion group, and bone marrow mesenchymal stem cells group. The rat models of primary nephrotic syndrome were established by single injection of adriamycin intravenously through tail vein in the latter two groups. Rats were then treated with bone marrow mesenchymal stem cells (1×10 7 ) (bone marrow mesenchymal stem cells group) or normal saline (normal saline infusion group) through tail vein at the same time after adriamycin administration. The normal group received no treatment. RESULTS AND CONCLUSION:Compared with the normal group, rats in the normal saline infusion group developed nephropathy characterized by ascites, proteinuria, hypoalbuminemia, hypercholastero-lnemia, and progressive renal injury. However, the proteinurine and clinical severity in bone marrow mesenchymal stem cells group were significantly ameliorated after treatment with bone marrow mesenchymal stem cells. CD4+CD25+Treg/CD4+Treg in the peripheral blood in the bone marrow mesenchymal stem cells group and normal saline infusion group were significantly higher than that in the normal group at 28

  12. Neuron-like cells differentiated from rat mesenchymal stem cells in vitro%体外诱导大鼠骨髓间充质干细胞向神经细胞分化的实验研究

    Institute of Scientific and Technical Information of China (English)

    李双月; 王哲敏; 陈若霖; 戚媛; 刘爽; 朴丰源

    2014-01-01

    目的:对大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells ,BMSCs)进行分离、培养与鉴定,并探讨全反式维甲酸(retinoic acid,RA)、碱性成纤维细胞生长因子(fibroblast growth factor,basic, bFGF)和表皮生长因子( epidermal growth factor , EGF )联合诱导BMSCs分化为神经细胞的可行性。方法全骨髓贴壁法分离培养BMSCs ,观察细胞形态及生长增殖情况;流式鉴定细胞表面标志物CD29、CD34、CD90;选用第3代细胞,经RA、bF-GF和EGF联合诱导后,细胞免疫化学染色检测神经细胞标志物神经元特异性烯醇化酶( neuron specific enolasen , NSE)的表达。结果体外培养的BMSCs呈成纤维细胞样,第3、4、5代BMSCs的生长曲线均呈S形,活性无明显差异。 BMSCs的均一性较好,第3代细胞CD29、CD90阳性率均在90%以上,而CD34阳性率仅为0.58%;BMSCs经诱导后分化为神经细胞,并表达神经细胞标志NSE。结论成功建立BMSCs 的体外培养体系,所得细胞纯度高、生物学特征稳定,并可诱导分化为神经细胞,为移植治疗神经系统损伤提供实验基础。%Objective To isolate and culture rat bone marrow mesenchymal stem cells (BMSCs) and induce them differen-tiate to neural cells .Methods BMSCs were isolated from rats by wall sticking method .Then the cells were identified with morphology , proliferation and the surface markers .Neuron-like cells from BMSCs were induced by the combination of ret-inoic acid(RA),fibroblast growth factor, basic (bFGF)and epidermal growth factor (EGF), and their neuron specific eno-lasen ( NSE ) were detected by NSE immunohistochemistry .Results Cultured BMSCs like fibroblast -shaped in vitro. Growth curves of the 3rd, 4th and 5th-generation BMSCs and their activities were not significantly different .FCM detected that CD29 and CD90 were positively expressed , but CD34 was negatively expressed .The induced

  13. Application of Bone Marrow Mesenchymal Stem Cells in Treatment of Rats with Spinal Cord Injuries%骨髓间充质干细胞在大鼠脊髓损伤治疗中的应用

    Institute of Scientific and Technical Information of China (English)

    薛荣利; 张媛媛; 李泽慧; 周丹; 张晨晨; 哈小琴

    2015-01-01

    Objective To investigate the effect of transplanting bone mesenchymal stem cells ( MSCs) in repair of rats with spinal cord injuries ( SCI) . Methods Bone marrow MSCs were isolated and cultured using whole bone mar-row cells adherence method, and models of SCI were established using free falling body hitting method. A total of 24 rats survived after model establishment, and were divided into transplantation group ( n=12 ) and control group ( n=12 ) . Transplantation group was transplanted with mesenchymal stem cells in injury sites, while control group was treated with same values of physiological saline. The recovery conditions of locomotory capabilities were evaluated using combined praxiology score, and the medullary histopathological changes, injuries and repair of nerve cells and expression of SRY gene were observed. Results The combined praxiology scores of transplantation group at different time points after the treatment were higher than those of control group (P<0. 05). In control group, there were cavities in spinal cord tissues and dead nerve fibers and cells. In transplantation group, the number of cavities was decreased, but hyperplastic tissues was increased uncreased as well as nerve fibers and cells, and SRY gene expression was found. Conclusion Transplan-tation of bone marrow MSCs may improve SCI repair.%目的 探讨骨髓间充质干细胞( MSCs)移植对大鼠脊髓损伤( SCI)修复的作用. 方法 全骨髓贴壁法分离培养骨髓MSCs,采用自由落体撞击法制作大鼠SCI模型,建模大鼠存活24只,分为移植组和对照组,每组12只,移植组在损伤部位移植培养的骨髓MSCs,对照组给予等量生理盐水. 根据联合行为学评分评价两组大鼠运动能力恢复情况,观察脊髓组织病理学变化、神经细胞损伤与修复及SRY基因的表达情况. 结果 移植组治疗后不同时间联合行为学评分均高于对照组(P<0. 05). 对照组脊髓组织中出现较多空腔,神经纤维及细胞

  14. Exosomes derived from miR-140-5p-overexpressing human synovial mesenchymal stem cells enhance cartilage tissue regeneration and prevent osteoarthritis of the knee in a rat model

    Science.gov (United States)

    Tao, Shi-Cong; Yuan, Ting; Zhang, Yue-Lei; Yin, Wen-Jing; Guo, Shang-Chun; Zhang, Chang-Qing

    2017-01-01

    OBJECTIVES: Osteoarthritis (OA) is the most common joint disease throughout the world. Exosomes derived from miR-140-5p-overexpressing synovial mesenchymal stem cells (SMSC-140s) may be effective in treating OA. We hypothesized that exosomes derived from SMSC-140 (SMSC-140-Exos) would enhance the proliferation and migration abilities of articular chondrocytes (ACs) without harming extracellular matrix (ECM) secretion. METHODS: SMSCs were transfected with or without miR-140-5p. Exosomes derived from SMSCs or SMSC-140s (SMSC-Exos or SMSC-140-Exos) were isolated and identified. Proliferation, migration and ECM secretion were measured in vitro and compared between groups. The mechanism involving alternative Wnt signalling and activation of Yes-associated protein (YAP) was investigated using lentivirus, oligonucleotides or chemical drugs. The preventative effect of exosomes in vivo was measured using Safranin-O and Fast green staining and immunohistochemical staining. RESULTS: Wnt5a and Wnt5b carried by exosomes activated YAP via the alternative Wnt signalling pathway and enhanced proliferation and migration of chondrocytes with the side-effect of significantly decreasing ECM secretion. Highly-expressed miR-140-5p blocked this side-effect via RalA. SMSC-140-Exos enhanced the proliferation and migration of ACs without damaging ECM secretion in vitro, while in vivo, SMSC-140-Exos successfully prevented OA in a rat model. CONCLUSIONS: These findings highlight the promising potential of SMSC-140-Exos in preventing OA. We first found a potential source of exosomes and studied their merits and shortcomings. Based on our understanding of the molecular mechanism, we overcame the shortcomings by modifying the exosomes. Such exosomes derived from modified cells hold potential as future therapeutic strategies. PMID:28042326

  15. In vivo differentiation of human amniotic epithelial cells into cardiomyocyte-like cells and cell transplantation effect on myocardial infarction in rats: comparison with cord blood and adipose tissue-derived mesenchymal stem cells.

    Science.gov (United States)

    Fang, Cheng-Hu; Jin, Jiyong; Joe, Jun-Ho; Song, Yi-Sun; So, Byung-Im; Lim, Sang Moo; Cheon, Gi Jeong; Woo, Sang-Keun; Ra, Jeong-Chan; Lee, Young-Yiul; Kim, Kyung-Soo

    2012-01-01

    Human amniotic epithelial cells (h-AECs), which have various merits as a cell source for cell therapy, are known to differentiate into cardiomyocytes in vitro. However, the ability of h-AECs to differentiate into cardiomyocytes in vivo and their cell transplantation effects on myocardial infarction are still unknown. In this study, we assessed whether h-AECs could differentiate into cardiomyocytes in vivo and whether h-AECs transplantation can decrease infarct size and improve cardiac function, in comparison to transplantation of cord blood-derived mesenchymal stem cells (MSCs) or adipose tissue-derived MSCs. For our study, we injected h-AECs, cord blood-derived MSCs, adipose tissue-derived MSCs, and saline into areas of myocardial infarction in athymic nude rats. After 4 weeks, 3% of the surviving h-AECs expressed myosin heavy chain, a marker specific to the myocardium. Compared with the saline group, all cell-implanted groups showed a higher ejection fraction, lower infarct area by positron emission tomography and histology, and more abundant myocardial gene and protein expression in the infarct area. We showed that h-AECs can differentiate into cardiomyocyte-like cells, decrease infarct size, and improve cardiac function in vivo. The beneficial effects of h-AECs were comparable to those of cord blood and adipose tissue-derived MSCs. These results support the need for further studies of h-AECs as a cell source for myocardial regeneration due to their plentiful availability, low immunity, and lack of ethical issues related to their use.

  16. Isolation,Cultivation and Identification of Bone Marrow Mesenchymal Stem Cells of Rats%大鼠骨髓间充质干细胞分离、培养及鉴定

    Institute of Scientific and Technical Information of China (English)

    王海峰; 陈文; 刘艳艳; 吴王泽

    2014-01-01

    目的:培养符合实验要求的大鼠骨髓间充质干细胞(bMSCs)并进行鉴定。方法采用全骨髓贴壁法培养大鼠 bMSCs,对细胞的形态及生长特性进行观察,并应用流式细胞术对细胞表面抗原 CD29、CD34、CD45、CD90表型进行鉴定。结果 bMSCs 呈贴壁生长,细胞状态良好,细胞形态以梭形为主,有时可见少量呈多角形,细胞平行排列生长或涡旋状生长;bMSCs 高表达干细胞表面抗原 CD29、CD90,而低表达造血干细胞表面抗原 CD34、CD45。结论全骨髓贴壁法培养 bMSCs 既简单又经济,通过多次传代可以达到细胞纯化的目的,对细胞的性状影响较小。%Objective To culture and identify the rat marrow mesenchymal stem cells(bMSCs)fitting the re-quirement of experimentation. Methods Whole bone marrow adherent culture method in vitro was used to culture the bMSCs of rats in vitro,the morphology and characteristics of the cells was observed,the phenotypes of CD29, CD34,CD45 and CD90 were identified by flow cytometry. Results bMSCs stuck wall growth in good condition, cells in a spindle primarily forms into fiber cell sample growth,sometimes visible small amounts of a polygonal cells,parallel arrangement growth or vortex shape growth. High expression of stem cell surface molecular(CD29 and CD90)and low expression of hematopoietic stem cell surface molecular(CD34 and CD45)were observed in the bM-SCs. Conclusion Whole bone marrow adherent culture method is simple and economical bMSCs culture method,the purified cells can be achieved through the passage,and the method has the little effect on cell trait.

  17. Bone marrow mesenchymal stem cells transplantation for treatment of dilated cardiomyopathy in rats%骨髓间充质干细胞移植治疗大鼠扩张型心肌病

    Institute of Scientific and Technical Information of China (English)

    徐燕; 张瑶; 李丽丽

    2012-01-01

    BACKGROUND: Myocardial fibrosis induced by dilated cardiomyopathy (DCM) is the pathological basis of heart failure. At present, drug treatment, interventionaI therapy and surgical intervention cannot ameliorate necrotic myocardium and completely improve cardiac function.OBJECTIVE: To investigate the effect of allogenic bone marrow mesenchymal stem cells (BMSCs) transplantation on cardiac function and myocardial fibrosis in rats with DCM.METHODS: Forty Wistar rats were randomly divided into three groups: cell transplantation group (n=15), control group (n=15) and blank group (n=10). DCM models were established in cell transplantation group and control group. Four weeks after the models were established successfully, rats in cell transplantation group were injected with allogeneic BMSCs 150 μL (containing 3×106 cells). Rats in control group and blank group were also implanted with the culture medium in the same amount. RESULTS AND CONCLUSION: Compared with the blank group, echocardiography showed that the left ventricular end-systolic inside diameter was increased before transplantation, while the ejection fraction and fractional shortening was decreased significantly (P < 0.01) in cell transplantation group and control group. Four weeks after transplantation, the echocardiography showed that the left ventricular end-systolic inner diameter was decreased, while the ejection fraction and fractional shortening was increased obviously (P < 0.01) in cell transplantation group compared with before transplantation. Expression of cardiac collagen in cell transplantation group was lower than that in control group (P < 0.05). Compared with the control group, the expression of matrix metalloproteinase 2 and matrix metalloproteinase 9 was significantly increased in the other two groups (P < 0.05). In conclusion, bone marrow mesenchymal stem cells (BMSCs) transplantation can improve cardiac function and ameliorate myocardial fibrosis in rats with DCM.%背景:扩张型心肌

  18. 骨髓间充质干细胞对大鼠到小鼠胰岛移植的保护作用%Bone Mesenchymal Stem Cells Induced lmmunotolerance on Rat to Mouse Islet Transplantation

    Institute of Scientific and Technical Information of China (English)

    曾力; 张爽; 魏玲玲; 田伯乐; 麦刚; 张杰; 李胜富; 陈又南; 陆燕蓉

    2011-01-01

    目的 探讨同种异基因骨髓间充质干细胞(bone mesenchamal stem cells,BMSC)静脉输注对大鼠到小鼠胰岛移植物的功能保护和小鼠糖尿病状态改善.方法 全骨髓培养法获得C57BL/6小鼠BMSC.不连续梯度离心法分离纯化Sprague-Dawley(SD)大鼠胰岛,将300胰岛当量的胰岛单独或与BMSC联合移植入链脲菌素诱导的糖尿病BALB/c小鼠肾包膜下,并通过尾静脉在移植后0.3和5d注射CM-DiI标记的BMSC 5×10(5)/只,对照组给于磷酸盐缓冲溶液.移植后监测血糖,第9天处死小鼠,取肝、脾、胸腺、淋巴结和移植胰岛的肾脏,冰冻切片,荧光显微镜观察CM-DiI标记细胞的组织分布;免疫荧光法观察移植物中胰岛素和胰高血糖素表达,评价胰岛的功能.结果 BMSC静脉输注后主要分布于胸腺,其次是脾脏和淋巴结,肾和肝组织中未观察到BMSC;BMSC联合胰岛移植组血糖控制水平优于其他组,且在第7天的口服糖耐量实验优于单纯胰岛移植组.结论 与胰岛联合移植的BMSC对受者免疫器官和组织有明显的趋向性,且对胰岛细胞的体内存活有一定保护作用.%Objective To research on the protection function by the allogeneic rat bone mesenchymal stem cells (BMSC) on rat to mouse islet transplantation and the improvement of diabetic state in muse. Methods BMSC were prepared from C57BL/6 mouse bone marrow cells and identified by flow cytometry (FCM). Islets were isolated from Sprague-Dawley (SD) rats with Ficoll discontinuous centrifugation. CM-DiI labeled BMSC at 5 × 105 for one mouse were intravenously infused into STZ induced diabetic BALB/c mice after rat to mouse islet transplantation at day 0, 3 and 5. Mice with PBS intravenously infused after islet transplantation were set as the negative controls. Blood glucose was monitored every day at the first 3 days after transplantation, and then monitored every two days. At day 9 after transplantation, spleen, thymus, lymph nods, liver and

  19. Differentiation of Bone Marrow Mesenchymal Stem Cells in Congenital Spina Bifida of Fetus Rats%骨髓间充质干细胞在先天性脊椎裂胎鼠脊髓中的分化

    Institute of Scientific and Technical Information of China (English)

    高飞; 袁正伟

    2012-01-01

    Objective To investigate the bone of survival and differentiation of mesenchymal stem cells ( MSC) in congenital spina bifida of fetus rats and to explore the theoretical clues for cell replacement of neural lube defects. Methods In sterile condition, rat bone marrow was flushed and cells were seeded in culture flask. Passage 3 of MSC were transfected by adenovirus enhanced green fluorescent protein ( ECFP) gene and then were injected into congenital spina bifida of fetus rats through fine glass needle. On 20 days of gestation, fetus rats underwent operation and spinal cords were removed for fixation and dehydration and got frozen sections. Expressions of Nestin( marker of neural stem cells) and glial fibrillary acidic protein(GFAP,marker of neurogliocyte) in transplanted MSC with EGKF were tested by immunofluores-cence. Results MSC surface antigen by flow cytometry showed CD34 ( - ) , CD44 occupying 95% , and CD90 occupying 99%. Transfection efficiency of adenovirus - EGFP to MSC was 61 %. The rate of dominant spina bifida was 64. 3% in teratogenic group. MSC distributed on the surface of spinal cord,or migrated into it. Some MSCs displayed different cell shapes such as long spindle or multiangle. Expressions of Nestin and GFAP were present in transplanted MSC by immunofluorescence assay. Conclusion MSC with EGFP can survive in fetus spinal cord after transplantation and differentiate into neural stem cells and neurogliocytes.%目的 观察大鼠骨髓间充质干细胞(MSC)在先天性脊椎裂胎鼠脊髓中的存活与分化,探寻细胞替代治疗先天性脊椎裂的方法.方法 用贴壁培养法进行大鼠MSC原代培养并传代,携带增强型绿色荧光蛋白(EGFP)基因的腺病毒转染第3代骨髓MSC.将转染的MSC注射到先天性脊椎裂胎鼠的脊髓中.母鼠孕20 d时取先天性脊椎裂胎鼠的脊椎,固定脱水后做冷冻切片,免疫荧光方法检测脊髓中EGFP阳性MSC中巢蛋白(Nestin)、胶质纤维

  20. Imprint Control of BaTiO3 Thin Films via Chemically Induced Surface Polarization Pinning.

    Science.gov (United States)

    Lee, Hyungwoo; Kim, Tae Heon; Patzner, Jacob J; Lu, Haidong; Lee, Jung-Woo; Zhou, Hua; Chang, Wansoo; Mahanthappa, Mahesh K; Tsymbal, Evgeny Y; Gruverman, Alexei; Eom, Chang-Beom

    2016-04-13

    Surface-adsorbed polar molecules can significantly alter the ferroelectric properties of oxide thin films. Thus, fundamental understanding and controlling the effect of surface adsorbates are crucial for the implementation of ferroelectric thin film devices, such as ferroelectric tunnel junctions. Herein, we report an imprint control of BaTiO3 (BTO) thin films by chemically induced surface polarization pinning in the top few atomic layers of the water-exposed BTO films. Our studies based on synchrotron X-ray scattering and coherent Bragg rod analysis demonstrate that the chemically induced surface polarization is not switchable but reduces the polarization imprint and improves the bistability of ferroelectric phase in BTO tunnel junctions. We conclude that the chemical treatment of ferroelectric thin films with polar molecules may serve as a simple yet powerful strategy to enhance functional properties of ferroelectric tunnel junctions for their practical applications.

  1. In vitro expression of the alpha-smooth muscle actin isoform by rat lung mesenchymal cells: regulation by culture condition and transforming growth factor-beta.

    Science.gov (United States)

    Mitchell, J J; Woodcock-Mitchell, J L; Perry, L; Zhao, J; Low, R B; Baldor, L; Absher, P M

    1993-07-01

    alpha-Smooth muscle actin (alpha SM actin)-containing cells recently have been demonstrated in intraalveolar lesions in both rat and human tissues following lung injury. In order to develop model systems for the study of such cells, we examined cultured lung cell lines for this phenotype. The adult rat lung fibroblast-like "RL" cell lines were found to express alpha SM actin mRNA and protein and to organize this actin into stress fiber-like structures. Immunocytochemical staining of subclones of the RL87 line demonstrated the presence in the cultures of at least four cell phenotypes, one that fails to express alpha SM actin and three distinct morphologic types that do express alpha SM actin. The proportion of cellular actin that is the alpha-isoform was modulated by the culture conditions. RL cells growing at low density expressed minimal alpha SM actin. On reaching confluent densities, however, alpha SM actin increased to at least 20% of the total actin content. This effect, combined with the observation that the most immunoreactive cells were those that displayed overlapping cell processes in culture, suggests that cell-cell contact may be involved in actin isoform regulation in these cells. Similar to the response of some smooth muscle cell lines, alpha SM actin expression in RL cells also was promoted by conditions, e.g., maintenance in low serum medium, which minimize cell division. alpha SM actin expression was modulated in RL cells by the growth factor transforming growth factor-beta. Addition of this cytokine to growing cells substantially elevated the proportion of alpha SM actin protein.(ABSTRACT TRUNCATED AT 250 WORDS)

  2. Plant flavonol isorhamnetin attenuates chemically induced inflammatory bowel disease via a PXR-dependent pathway

    OpenAIRE

    Dou, Wei; Zhang, Jingjing; Li, Hao; Kortagere, Sandhya; Sun, Katherine; Ding, Lili; Ren, Gaiyan; Wang, Zhengtao; Mani, Sridhar

    2014-01-01

    Isorhamnetin is an O-methylated flavonol present in fruit and vegetables. We recently reported the identification of isorhamnetin as an activator of the human pregnane X receptor (PXR), a known target for abrogating inflammation in inflammatory bowel disease (IBD). The current study investigated the role of isorhamnetin as a putative mouse PXR activator in ameliorating chemically induced IBD. Using two different models (Ulcerative colitis-like and Crohn’s disease-like) of experimental IBD in ...

  3. 棕色脂肪干细胞与白色脂肪干细胞移植对心肌梗死大鼠心功能的影响%Effects of brown adipose-derived mesenchymal stem cells and white adipose-derived mesenchymal stem cells transplantation on cardiac function of rat with myocardial infarction

    Institute of Scientific and Technical Information of China (English)

    石金鑫; 刘剑锋; 王海滨; 朱平

    2013-01-01

    Objective To compare the therapeutic effects of brown adipose-derived mesenchymal stem cells (BADSCs)and white adipose-derived mesenchymal stem cells (ADSCs) in rats with acute myocardial infarction,Methods The BADSCs and ADSCs were prepared by enzyme digestion from brown and white adipose tissues isolated from rat shoulder and inguinal regions,respectively,The stem ceils were analyzed by flow cytometry and multipotential differentiation ability.30 rats were randomly divided into PBS control group,ADSCs transplantation group and BADSCs transplantation group,then the acute myocardial infarction model was established.The BADSCs or ADSCs were injected into the edge area of myocardial infarction respectively,the PBS was injected as control.Detailed histological analysis and echocardiography were used to determine the cardiac function and differentiation,vascularization effects of transplantation cells after 4 weeks of transplantation.Results Compared with PBS control group,the heart function improved significantly with decreased fibrosis formation in the BADSCs and ADSCs transplantation at 4 weeks after transplantation (P < 0,05),Immunofluorescence result showed that the cardiac-differentiation ability of BADSCs was much higher than that of ADSCs in vivo,Immunohistochemical results showed that compared with the PBS control group,the micro-vessel density in scar areas increased significantly in the ADSCs transplantation group and BADSCs transplantation group (P < 0,05),and the ADSCs groups increased most (P < 0.05),Conclusion Both transplantation of BADSCs and ADSCs can enhance the cardiac function of myocardial infarction.Although compared with ADSCs,BADSCs has a lower ability of vascularization,but it can improve the heart performance through the high potential of cardiac-differentiation.%目的 对比研究棕色脂肪干细胞(BADSCs)与白色脂肪干细胞(ADSCs)治疗急性心肌梗死大鼠的效果.方法 采用酶消化法分别分离大鼠腹股沟脂

  4. Intravenous administration of mesenchymal stem cells overexpressing CXCR4 protects against experimental colitis in rats%过表达CXCR4的间充质干细胞缓解实验性结肠炎

    Institute of Scientific and Technical Information of China (English)

    刘星星; 范恒; 唐庆; 寿折星; 陶玲; 张丽娟; 左冬梅

    2016-01-01

    目的:观察过表达CXCR4的骨髓间充质干细胞治疗2,4,6-三硝基苯磺酸诱导的实验性结肠炎的效果及其潜在的免疫作用机制.方法:从♀SD大鼠骨髓中分离骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)并使用流式细胞术鉴定.通过慢病毒技术使BMSCs表达GFP(green fluorescent protein-GFP,Ad-GFP-BMSCs)或共表达CXCR4和GFP(Ad-CXCR4-BMSCs),32只大鼠被随机分成4组(n=8):空白组、模型组、Ad-GFP-BMSCs组、Ad-CXCR4-BMSCs组.采用TNBS诱导实验性结肠炎,尾静脉注射Ad-CXCR4-BMSCs或Ad-GFP-BMSCs.治疗1 wk,收集结肠组织进行HE染色和病理学分析.PCR检测结肠部位干扰素-γ(interferon-γ,IFN-γ)、肿瘤坏死因子-α(tumor necrosis factor α,TNF-α)、白介素(interleukin,IL)-6和IL-10的mRNA表达,Western blot检测信号传导子及转录激活子(signal transducer and activator of transcription,STAT)-3和磷酸化STAT-3蛋白表达,免疫组织化学检测磷酸化STAT3蛋白表达.结果:相对于空白组,模型组结肠部位的IFN-γ、TNF-α、IL-6和IL-10的mRNA表达显著上升,S TAT-3及磷酸化STAT-3的蛋白表达明显上升(P<0.05).系统治疗1 wk后,相对于模型组,Ad-GFP-BMSCs组大鼠的临床症状及结肠病理损害并没有得到有效的缓解.相对于Ad-GFP-BMSCs组,Ad-CXCR4-BMSCs组大鼠结肠组织的IFN-γ、TNF-α、IL-6的mRNA表达显著下降,IL-10 的mRNA表达显著上升(P<0.05);STAT3 及磷酸化STAT-3的蛋白表达显著下降(P<0.05).结论:过表达CXCR4的BMSCs可能通过抗炎及免疫调节机制来缓解实验性结肠炎,这可能为BMSC缓解IBD提供可靠的理论依据.%AIM:To investigate the role of SDF-1α/CXCR4 axis in the therapeutic effects of lentivirus-preconditioned bone mesenchymal stem cells (BMSCs) for 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis in rats.METHODS:BMSCs were isolated from Sprague-Dawley (SD) rats and identified by flow cytometry.Lentivirus transfection was applied to

  5. 骨髓间充质干细胞移植抗大鼠肝纤维化的作用及机制%Effect of Mesenchymal Stem Cells Transplantation on Anti-Hepatic Fibrosis in Rats and Its Mechanism

    Institute of Scientific and Technical Information of China (English)

    陶霖; 姜海行; 覃山羽; 金群馨; 黄雪; 郭晓云; 谭至柔; 唐国都

    2014-01-01

    Objective To study the effect of mesenchymal stem cells ( MSCs ) transplantation on rats with hepatic fibrosis and its mechanism.Methods MSCs was isolated from Sprague-Dawley(SD) rats and cultured for 4 passages.The hepatic fibrosis model of SD rats was induced by subcutaneous injection of CCl 4.Sixty SD hepatic fibrosis rats were divided into 3 groups randomly (20 rats in each group):①Control group:The rats were treated by infusion of normal saline via tail vein.②MSCs group:The rats were treated by injection of MSCs suspension via tail vein .③Induced group:The rats were treated by injection of hepatocyte growth factor (HGF)-induced MSCs suspension via tail vein 14 days after induction.The rats were killed 1 week,2 weeks,3 weeks,4 weeks after MSCs transplantation,respectively,5 rats for each batch.The serum levels of hyaluronic acid (HA),laminin(LN),collagen Ⅳ(Col-Ⅳ) were detected.The degree of hepatic fibrosis was observed with light microscope.The mRNA expressions of collagenⅠ(Col-Ⅰ),RhoA,Cdc42,ras-related C3 botulinum toxin substrate(Rac1) in hepatic tissues were detected by PCR ,whereas the protein expressions of Col-Ⅰ,RhoA,Cdc42,Rac1 in hepatic tissues were detected by Western blot method .Results ①The score of hepatic fibrosis degree was smaller with increasing time in the induced group and MSCs group ,which was smaller in the induced group in the fourth week in contrast with that in the MSCs group (P<0.05).②The serum levels of HA,LN,Col-Ⅳdecreased significantly in each group 3 weeks after transplantation ,and they were significantly lower in the induced group ,MSCs group compared with control group ,which were lower in the induced group in contrast with those in the MSCs group ( P<0 .05 ) .③After MSCs transplantation,the mRNA and protein expressions of Col-Ⅰ,RhoA,Cdc42,Rac1 in hepatic tissues reduced more signif-icantly in the MSCs group and induced group with increasing time ,which were significantly lower in the induced group 4

  6. 间充质干细胞移植减轻大鼠盐酸吸入性肺损伤%Bone marrow-derived mesenchymal stem cells alleviate lung injury in a rat model of acid aspiration

    Institute of Scientific and Technical Information of China (English)

    吴晓丹; 贾庆安; 钱梦佳; 隆玄; 李善群; 宋元林; 王向东; 白春学

    2013-01-01

    目的 研究骨髓间充质干细胞(BMSCs)移植对大鼠盐酸吸入性肺损伤的作用与机制.方法 本实验于复旦大学附属中山医院实验研究中心进行.改良贴壁法分离培养Sprague-Dawley (SD)大鼠BMSCs.24只SD大鼠随机(随机数字法)分为3组(n=8):对照组,损伤组和移植组.损伤组和移植组气道内滴注盐酸(1.2 ml/kg,pH=1.5),对照组则滴入等量磷酸缓冲盐溶液(PBS),用动脉血气、肺组织湿干比和病理改变评价模型是否成功建立.移植组于颈外静脉注射5×106个BMSCs,其他2组注入0.5 ml的PBS.移植6h后行动脉血气分析,酶联免疫吸附法(ELISA)测定血清和支气管肺泡灌洗液(BALF)的肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)和白介素-10 (IL-10)质量浓度,观察肺组织病理改变和测定肺组织湿干比.体外实验中,BMSCs与损伤肺细胞或正常肺细胞于Transwell体系共培养36 h,观察损伤肺影响BMSCs迁移情况;另取BMSCs与损伤肺细胞昆合培养或于Transwell中培养,正常肺细胞或损伤肺细胞单独培养作对照,6h后检测上清液中TNF-α、IL-6和IL-10质量浓度.用方差分析比较多组间的差异,独立样本t检验比较两组间差异.结果 BMSCs移植改善了肺损伤大鼠的低氧血症(P<0.01)和肺组织损伤,降低了肺组织湿干比(P<0.01),降低了血清和BALF的TNF-α水平(P<0.05;P<0.01),升高了血清和BALF的IL-10质量浓度(P<0.01),对IL-6水平无明显影响(P>0.05).体外实验中,BMSCs向损伤肺迁移明显多于向正常肺迁移(P<0.01),BMSCs与肺细胞接触或不接触的共培养均降低了上清液TNF-α质量浓度(P<0.01),升高了IL-10质量浓度(P<0.01),对IL-6质量浓度无影响(P>0.05).结论 BMSCs移植可以减轻大鼠盐酸吸入性肺损伤,可能机制为干细胞通过旁分泌途径下调炎症反应.%Objective To investigate the effects and the possible mechanism of bone mesenchymal stem cells (BMSCs) transplantation in

  7. 骨髓间充质干细胞在全身照射大鼠体内分布实验研究%The distribution of mesenchymal stem cells after total-body irradiation in rats

    Institute of Scientific and Technical Information of China (English)

    白守民; 梁碧玲; 李志伟; 韩非; 陈春燕; 卢泰祥

    2009-01-01

    Objective To detect the distribution of mesenchymal stem cells(MSCs) after total-body irradiation in rats. Methods MSCs were cultured and labeled with green fluorescent protein(GFP). Rats were exposed to total-body irradiation(TB1) or TBI plus total brain irradiation, and then MSCs were injected through the tail vein. The Fluorescent MSCs were observed by fluorescence microscope. The MSCs numbers in different organs were determined by quantitative RT-PCR method. Results GFP-labeled MSCs were ob-tained. After MSCs were infused to the rats,few of them were observed in the organs of nonirradiated group except for a very low number in the lungs,bone marrow(BM) and spleen. TBI of 6 Gy increased the engraft-ment of MSCs in almost all the organs, especially in early response tissues such as the small intestine and BM. TBI of 7 Gy further increased the number of MSCs. The MSCs numbers in the brain and other organs were significantly increased after 20 Gy total brain irradiation in addition to 6 Gy TBI. Conclusions Radi-ation injury can induce the aggregation of MSCs. With the increase of radiation dose and severity of radiation injury,a significant increase of MSCs in different organs were observed. Local irradiation can increase the MSCs distribution in the radiation field as well as other organs.%目的 用荧光定量RT-PCR方法探讨骨髓间充质干细胞在大鼠经过全身照射后其体内分布的状况.为进一步研究骨髓间充质干细胞(MSCs)对正常组织器管放射性损伤的治疗提供理论依据.方法 培养MSCs细胞,用绿色荧光蛋白标记.分别对大鼠行全身照射及全身照射+全脑照射,经尾静脉注入标记的MSCs,荧光显微镜观察MSCs在各器官的分布,荧光定量RT-PCR检测MSCs在不同组织器官的分布.结果 获得绿色荧光标记MSCs,经尾静脉注入各实验组大鼠体内后显示,未行照射时MSCs仅少量分布在肺、骨髓、脾中;6 Gy全身照射后各器官MSCs的分布明显增加,小

  8. Peripheral nerve extract effects on mesenchymal cells.

    Science.gov (United States)

    Dietz, F R; Mukhopadhyay, B; Becker, G; Daniels, K; Solursh, M

    1996-01-01

    Several common congenital limb disorders are characterized by normal tissue differentiation but abnormal somatic growth. These include: idiopathic clubfoot, idiopathic leg length discrepancy, hemi-atrophy and hemi-hypertrophy. Both clinical and research studies have suggested that peripheral nerves may be important in regulating somatic growth of limb tissues. To investigate the hypothesis that peripheral nerves convey trophic substances to mesenchymal tissues that are involved in the regulation of growth, we developed an in vitro assay to assess the effect of fractions of peripheral nerve on myoblast and chondroblast growth and differentiation in a mammalian (rat) system. Whole rat sciatic nerve extract was fractionated by ammonium sulfate precipitation and by affinity chromatography. Concavalin A chromatography resolved whole nerve extract into a glycoprotein and a non-glycoprotein fraction. Serial ammonium sulfate precipitation yielded three pellet fractions designated as 35%, 70%, and 100% pellets; corresponding to ammonium sulfate concentrations of 0 to 35%, 35 to 70%, and 70 to 100% saturation, respectively. Dialyzed solutions of these pellets as well as the fractions from Concavalin A chromatography were assayed for biological activity in micromass cultures of rat limb bud mesenchyme, which allowed assessment of both myoblast and chondroblast stimulation. Stimulation of protein synthesis and myoblast proliferation (as measured by MF20 staining) occurred with both 70% and 100% ammonium sulfate fractions. Stimulation of chondroblasts (as measured by the number of alcian blue staining nodules) occurred with the 35% and 100% fractions. The glycoprotein fraction from the affinity chromatography stimulated protein synthesis and myoblast proliferation and inhibited chondroblast development. Stimulation of chondroblasts was seen with the non-glycoprotein fraction. No effect on protein synthesis, myoblast proliferation or chondroblast proliferation was found in

  9. Infusion of mesenchymal stem cells limits fibrogenesis in irradiated rat lung%间充质干细胞对延缓大鼠放射性肺纤维化进展的作用研究

    Institute of Scientific and Technical Information of China (English)

    常鹏宇; 夏诚诚; 侯雪; 石硙岩; 宋宇哲; 张玉宇; 赵钦; 马利新; 曲雅勤

    2015-01-01

    Dawley rats were enrolled in this study.For establishing the model of radiation-induced lung injury,36 rats were randomly selected to administrate a dose of 15 Gy to their whole right lungs.Thereafter,the irradiated rats were randomly classified into three groups.Each group contained 12 rats.Phosphate buffered sodium (PBS group),human skin-derived fibroblasts (fibroblast group) and human adipose tissue-derived mesenchymal stem cells (Ad-MSC group) were injected into rat through the tail vein respectively.The rest 12 rats without semi-thoracic irradiation were set as healthy controls.At 24 weeks post-irradiation,radiological imaging and pathological analyses were used for evaluating the histological changes in irradiated lungs.The pathological analyses included H&E staining for reflecting pulmonary architecture,Masson's Trichrome (M.T) staining for evaluating extracellular matrix (ECM) formation and IHC-staining for identifying the expressions of alpha-smooth muscle actin (α-SMA) and TGF-β1 in irradiated lungs.Additionally,the concentrations of HGF and TGF-β1 in serum and BALF were tested by ELISA assay.Quantitative real-time PCR assay was used to test the gene-expressions in irradiated lungs,including Collagen Ⅰ-α1 and Collagen Ⅲ-α1.Results The chest radiological imaging revealed that high-density shadows were presented in the irradiated lungs at 24 weeks post-irradiation.The lower lobes were predominant disease.However,the lesion of high-density shadow was not obvious in Ad-MSC group.Histological analysis by H&E staining indicated that the irradiated lungs in both PBS and fibroblast groups lost their architectures,representing by alveolar collapse and thickening alveolar septa.Moreover,compared to Ad-MSC group,excessive ECM formation together with significantly up-regulated expressions of Collagen Ⅰ-α1 and Collagen Ⅲ-α1 (F =4.39,7.73,P < 0.05) were also found within the injured tissue treated with PBS or fibroblasts.Apart from this,obvious expressions of

  10. Postnatal epithelium and mesenchyme stem/progenitor cells in bioengineered amelogenesis and dentinogenesis.

    Science.gov (United States)

    Jiang, Nan; Zhou, Jian; Chen, Mo; Schiff, Michael D; Lee, Chang H; Kong, Kimi; Embree, Mildred C; Zhou, Yanheng; Mao, Jeremy J

    2014-02-01

    Rodent incisors provide a classic model for studying epithelial-mesenchymal interactions in development. However, postnatal stem/progenitor cells in rodent incisors have not been exploited for tooth regeneration. Here, we characterized postnatal rat incisor epithelium and mesenchyme stem/progenitor cells and found that they formed enamel- and dentin-like tissues in vivo. Epithelium and mesenchyme cells were harvested separately from the apical region of postnatal 4-5 day rat incisors. Epithelial and mesenchymal phenotypes were confirmed by immunocytochemistry, CFU assay and/or multi-lineage differentiation. CK14+, Sox2+ and Lgr5+ epithelium stem cells from the cervical loop enhanced amelogenin and ameloblastin expression upon BMP4 or FGF3 stimulation, signifying their differentiation towards ameloblast-like cells, whereas mesenchyme stem/progenitor cells upon BMP4, BMP7 and Wnt3a treatment robustly expressed Dspp, a hallmark of odontoblastic differentiation. We then control-released microencapsulated BMP4, BMP7 and Wnt3a in transplants of epithelium and mesenchyme stem/progenitor cells in the renal capsule of athymic mice in vivo. Enamel and dentin-like tissues were generated in two integrated layers with specific expression of amelogenin and ameloblastin in the newly formed, de novo enamel-like tissue, and DSP in dentin-like tissue. These findings suggest that postnatal epithelium and mesenchyme stem/progenitor cells can be primed towards bioengineered tooth regeneration.

  11. Repair of calvarial defects with human umbilical cord blood derived mesenchymal stem cells and demineralized bone matrix in athymic rats%人脐血间充质干细胞修复颅骨缺损的实验研究

    Institute of Scientific and Technical Information of China (English)

    刘广鹏; 李宇琳; 孙剑; 崔磊; 张文杰; 曹谊林

    2010-01-01

    Objective To investigate the feasibility of using human umbilical cord blood derived mesenchymal stem cells (UCB-MSCs) and demineralized bone matrix (DBM) scaffolds to repair critical-sized calvarial defects in athymic rats. Methods Human UCB-MSCs were isolated, expanded and osteogenically induced in vitro. Osteogenic differentiation of UCB-MSCs was evaluated by Alizarin Red staining and measurement of calcium content respectively, and then the cells were seeded onto DBM scaffolds. Bilateral full-thickness defects (5 mm in diameter) of parietal bone were created in an athymic rat model. The defects were either repaired with UCB-MSC/DBM constructs (experimental group) or with DBM scaffolds alone (control group). Animals were harvested at 6 and 12 weeks post-implantation respectively, and defect repair was evaluated with gross observation, micro-CT measurement and histological analysis. Results Micro-CT showed that new bone was formed in the experimental group at 6 weeks post-implantation, while no sign of new bone formation was observed in the control group. At 12 weeks post-transplantation, scaffolds had been degraded almost completely in both sides. It was shown that an average of (78.19±6.45)% of each defect volume had been repaired in experimental side; while in the control side, only limited bone formed at the periphery of the defect. Histological examination revealed that the defect was repaired by trabecular bone tissue in experimental side at 12 weeks, while only fibrous connection was observed in the control group. Conclusions Tissue-engineered bone composed of osteogenically-induced human UCB-MSCs on DBM scaffolds could successfully repair the critical-sized calvarial defects in athymic rat models.%目的 应用人脐血间充质干细胞(umbilical cord blood derived mesenchymal stem cells,UCB-MSCs)复合脱钙骨材料构建组织工程化骨,修复裸大鼠颅骨标准缺损.方法 体外扩增培养、成骨诱导人UCB-MSCs,采用Alizarin Red染色

  12. 骨髓间充质干细胞移植在肺气肿大鼠模型肺组织内定植研究%Study on the Bone Marrow Mesenchymal Stem Cell Transplantation in Rats Lung Tissue with Emphysema

    Institute of Scientific and Technical Information of China (English)

    何桦; 赵祝香; 张颖

    2014-01-01

    目的:观察骨髓间充质干细胞移植在肺气肿大鼠模型肺组织内的定植情况。方法:选择健康SD大鼠34只,按随机数字表法分为MSCs干预组(A组,10只,慢阻肺大鼠,尾静脉输注MSC 1×106个/mL),肺气肿模型组(B组,10只,慢阻肺大鼠,尾静脉输注等体积PBS)及MSC对照组(C组,10只,正常大鼠,尾静脉输注MSCs 1×106个/mL),正常对照组(D组,4只,正常大鼠,尾静脉输注等体积PBS),采用烟熏法复制大鼠肺气肿模型。全骨髓培养法体外培养扩增雄性SD大鼠来源的MSCs,经GFP标记细胞后将其经尾静脉注入肺气肿模型SD大鼠体内,24 h内处死大鼠,取肺组织迅速冰冻切片,共聚焦激光显微镜下观察观察转染GPF的间充质干细胞在大鼠肺内定植情况。结果:成功培养具有分化潜能的骨髓间充质干细胞,MSCs传至第4代时有99.5%表达CD44、99.6%表达CD29等间充质干细胞表面标志,仅有0.4%表达CD34、1.0%表达CD45单核细胞以及造血干细胞表型;成功复制大鼠肺气肿模型,香烟烟雾暴露组(A、B组)平均肺泡间隔为(119.0±26.2)µm,高于对照组(C、D组)的(89.8±17.3)µm,差异有统计学意义(P<0.05);平均肺泡数为(173.9±68.3)个/mm2低于对照组的(280.3±104.0)个/mm2,差异有统计学意义(P<0.05);显微共聚焦发现MSCs经尾静脉注入大鼠体内24 h后可见A组大鼠肺组织内转染绿色荧光蛋白质粒的MSCs,而B组、C组及D组均未见转染荧光。结论:骨髓间充质干细胞经尾静脉输注入后可在肺气肿模型大鼠肺内定植,为MSCs治疗慢阻肺可能提供理论依据。%To observe the situation of bone marrow mesenchymal stem cell transplantation in rats lung tissue with emphysema.Method:34 healthy rats were randomly divided into the MSCs intervention group(the group A,10 COPD rats,tail intravenous MSC 1×106cells

  13. Adipose-derived mesenchymal stem cell therapy for radiation-induced vascular injury in small intestine of rat%脂肪干细胞修复辐射诱导的肠血管损伤研究

    Institute of Scientific and Technical Information of China (English)

    常鹏宇; 崔爽; 姜新; 曲超; 蒋鑫萍; 罗景华; 曲雅勤; 董丽华

    2014-01-01

    目的:评价人源脂肪干细胞对辐射诱导的肠血管损伤的修复作用。方法选用成年雄性SD大鼠,共34只,给予全腹部15 Gy X射线照射。造模后,取其中17只给予P6代人源脂肪干细胞腹腔注射治疗( Ad-MSC治疗组);另17只大鼠给予双磷酸盐缓冲液( PBS)腹腔注射治疗( PBS溶剂对照组)。照射后第10天,流式细胞分析绒毛内CD31阳性内皮细胞的数量,免疫荧光染色分析新生的血管内皮细胞,免疫组织化学染色分析血管结构的连续性;并提取受照小肠组织总RNA,实时荧光定量PCR检测受照组织内基质细胞起源因子-1(SDF-1)、血管内皮细胞生长因子(VEGF)、碱性成纤维细胞生长因子(bFGF)以及血管内皮细胞生长因子受体(Flk-1)的表达量。检测Ad-MSC治疗组的受损小肠内新生的内皮细胞来源。结果与PBS溶剂对照组相比,Ad-MSC能够显著增加受损组织内CD31阳性内皮细胞数量(t=12�15,P<0�05),提高受损组织内的血管密度(20 d:t=10�33,P<0.05;30 d:t=32�85,P<0�05),上调血管生成基因VEGF、bFGF、Flk-1以及SDF-1的表达量(t=10�34、11�25、6�73、6�73,P<0�05)。维持受损部位小肠绒毛内的血管完整,并促进CD31阳性造血干/祖细胞向血管内皮细胞的分化,加速受损部位血管的新生。结论人源脂肪干细胞通过发挥促血管新生的作用来修复辐射诱导的肠血管损伤。%Objective To assess the therapeutic effect of human adipose-derived mesenchymal stem cells on radiation-induced vascular injury in the small intestine of rat. Methods A total of 34 male Sprague-Dawley rats were enrolled in this study. To establish a model of radiation-induced intestinal injury, each rat was irradiated with 15 Gy in whole abdomen. 17 rats were randomly selected and infused intraperitoneally with passage 6 ( P6 ) Ad-MSCs, and the other 17

  14. Role of Wnt/β-catenin signaling in aging of mesenchymal stem cells of rats%Wnt/β-catenin信号通路对问充质干细胞衰老的影响及其作用机制

    Institute of Scientific and Technical Information of China (English)

    项晓霞; 陈律; 王骏浩; 张余斌; 张大勇

    2011-01-01

    Objective: To investigate the role of Wnt/β-catenin signaling in aging of mesenchymal stem cells (MSCs) of rats. Methods: Serum samples were collected from young (8 ~ 12 w) and aged (64 ~72 w) SD rat. Four experiment groups were assigned:young rat serum ( YRS) , YRS + Wnt 3a,old rat serum ( ORS) and ORS + DKK1 groups. Immunofluorescence and Western blotting were used to detect the expression of intracellular (3-catenin. The senescence-associated changes were examined with SA-|3-galactosidase staining. The proliferation ability was tested by MTT assays. The survived and apoptotic cells were determined by AO/EB staining. The expressions of γ-H2A. X and p53 protein were detected by immunofluorescence and Western blotting. RT-PCR was used to detect the expression of p53 and p21 mRNA. Results: Compared with the YRS group,the intracellular expression of (3-catenin in the ORS group was significantly increased, especially in the nuclei of MSCs. After treatment of DKK1 in ORS, the (3-catenin expression was reduced. The number of SA-|3-galactosidase positive MSCs was significantly higher in the YRS + Wnt 3a group than that in the YRS group ( P < 0. 01 ) , and the proliferative and survival ability of MSCs was significantly decreased in the YRS + Wnt 3a group. The number of SA-|3 -galactosidase positive MSCs in the ORS + DKK1 group was significantly decreased compared with that in ORS group (P<0.01 ) ,and the proliferative and survival ability of MSCs was significantly increased in the ORS + DKK1 group. The expression of γ-H2A. X,p53 and p21 was markedly increased in the ORS group than that in YRS group,however,after treatment with Wnt/β-catenin signaling inhibitor DKKl,the expression of 7-H2A. X,p53 and p21 was significantly decreased compared with that in the ORS group. Conclusions; Results suggest that the Wnt/p-catenin signaling is activated in the MSCs cultured with ORS and excessive activation of Wnt/p-catenin signaling can promote MSCs aging. The DNA damage

  15. Systems toxicology of chemically induced liver and kidney injuries: histopathology-associated gene co-expression modules.

    Science.gov (United States)

    Te, Jerez A; AbdulHameed, Mohamed Diwan M; Wallqvist, Anders

    2016-09-01

    Organ injuries caused by environmental chemical exposures or use of pharmaceutical drugs pose a serious health risk that may be difficult to assess because of a lack of non-invasive diagnostic tests. Mapping chemical injuries to organ-specific histopathology outcomes via biomarkers will provide a foundation for designing precise and robust diagnostic tests. We identified co-expressed genes (modules) specific to injury endpoints using the Open Toxicogenomics Project-Genomics Assisted Toxicity Evaluation System (TG-GATEs) - a toxicogenomics database containing organ-specific gene expression data matched to dose- and time-dependent chemical exposures and adverse histopathology assessments in Sprague-Dawley rats. We proposed a protocol for selecting gene modules associated with chemical-induced injuries that classify 11 liver and eight kidney histopathology endpoints based on dose-dependent activation of the identified modules. We showed that the activation of the modules for a particular chemical exposure condition, i.e., chemical-time-dose combination, correlated with the severity of histopathological damage in a dose-dependent manner. Furthermore, the modules could distinguish different types of injuries caused by chemical exposures as well as determine whether the injury module activation was specific to the tissue of origin (liver and kidney). The generated modules provide a link between toxic chemical exposures, different molecular initiating events among underlying molecular pathways and resultant organ damage. Published 2016. This article is a U.S. Government work and is in the public domain in the USA. Journal of Applied Toxicology published by John Wiley & Sons, Ltd.

  16. Improving analytical methods for protein-protein interaction through implementation of chemically inducible dimerization

    DEFF Research Database (Denmark)

    Andersen, Tonni Grube; Nintemann, Sebastian; Marek, Magdalena;

    2016-01-01

    into the widely used split ubiquitin-, bimolecular fluorescence complementation (BiFC)- and Forster resonance energy transfer (FRET)-based methods and investigated different protein-protein interactions in yeast and plants. We demonstrate the functionality of this concept by the analysis of weakly interacting...... proteins from specialized metabolism in the model plant Arabidopsis thaliana. Our results illustrate that chemically induced dimerization can function as a built-in control for split-based systems that is easily implemented and allows for direct evaluation of functionality....

  17. Improving analytical methods for protein-protein interaction through implementation of chemically inducible dimerization

    DEFF Research Database (Denmark)

    Andersen, T. G.; Nintemann, S. J.; Marek, M.;

    2016-01-01

    When investigating interactions between two proteins with complementary reporter tags in yeast two-hybrid or split GFP assays, it remains troublesome to discriminate true-from false-negative results and challenging to compare the level of interaction across experiments. This leads to decreased...... sensitivity and renders analysis of weak or transient interactions difficult to perform. In this work, we describe the development of reporters that can be chemically induced to dimerize independently of the investigated interactions and thus alleviate these issues. We incorporated our reporters...

  18. 骨髓间充质干细胞移植对实验性自身免疫性葡萄膜炎的抑制作用%Transplantation of bone marrow mesenchymal stem cells inhibits rat experimental autoimmune uveitis

    Institute of Scientific and Technical Information of China (English)

    解传奇; 李静; 贾亚丁

    2010-01-01

    Objective To investigate the impact of bone marrow mesenchymal stem cell transplantation on a rat model of experimental autoimmune uveitis (EAU) and analyze its immune regulatory mechanisms in vivo. Methods Eighteen Lewis rats were randomly divided into three groups: model control group, intervention group and normal control group, six animals in each group. Human retinal S-antigen peptide (HS-AgP35, 1 mg/ml) was mixed and emulsified with complete Freund's adjuvant and injected into hind foot pad of rats on the first and eighth day to establish the animal model of EAU. For bone marrow mesenchymal stem cell transplantation, 1 ml of cell suspension (2 × 106 cells/ml) was injected into tail vein of the intervention group rats on the first day when the emulsified S-antigen was injected. EAU manifestation, pathological change and IFN-γ level were evaluated and compared among those three groups after two weeks. Results No abnormal signs were found in the eyes of rats in normal control group. The manifestation grading of the intervention group (two rats at grade 0, three rats at grade 0.5, one rat at grade one) was significantly different from the model control group (one rat at grade one, one rat at grade two, three rats at grade three, one rat at grade four) (P=0. 015). The retina of rats in normal control group was ordinary under light microscope. The histopathological grading of the intervention group (one rat at grade 0, four rats at grade 0.5, one rat at grade one) and the model control group (four rats at grade three, two rats at grade four) was also statistically different (P<0. 01). Furthermore, the IFN-γ level in peripheral blood of the intervention group rats declined significantly compared to the model control group (t=9. 057 4, P=0. 01). Conclusions Bone marrow mesenchymal stem cells can inhibit EAU significantly,possibly by lowering the level of IFN-γ, thereby reduce the severity of uveitis and improve the condition of uveitis in rats.%目的 观察

  19. 骨髓间充质干细胞移植促进大鼠子宫创伤修复%Bone marrow mesenchymal stem cells transplantation promotes regeneration repair of uterine incision in rat

    Institute of Scientific and Technical Information of China (English)

    韦伟; 郑飞云; 杨孝军

    2012-01-01

    Objective To investigate the growth and differentiation of adenovirus-green fluorescent protein labelled bone marrow mesenchymal stem cells (BM-MSCs) in uterus incision, and to discuss its potential regeneration repair effect on uterine incision in rat. Methods BM-MSCs were collected by density gradient centrifugation and cultured for fourth generations, then transfected and labelled with adenovirus-green fluorescent protein. Fourty female SD rats were divided equally into control and experimental group, followed with the establishment of uterine wound model. After two weeks, adenovirus-green labelled BM-MSCs were transplanted directly into uterus at previous incision, with the control group only received normal saline injection. Then, one month later, CTGF, CD34 and Masson were used to detect the outcome of the transplanted BM-MSCs. Results The transplanted BM-MSCs can growth well and showed some extent of differentiation toward smooth muscle cells. Significant difference existed for CTGF and Masson between experimental and control samples (both P < 0. 01). Conclusions BM-MSCs transplantation is a promising technique and can be used to promote regeneration repair of uterine incision.%目的 探讨移植骨髓间充质干细胞移植(BM-MSCs)后在子宫切口部位的生长分化及其在大鼠子宫创伤后再生修复中的作用.方法 分离和培养大鼠BM-MSCs,传代扩增至第4代利用腺病毒-绿色荧光蛋白感染干细胞并继续培养.取雌性SD大鼠40只,分为对照组及实验组,建立子宫损伤动物模型,实验组移植BM-MSCs,对照组注射0.9%氯化钠注射液,2周后观察干细胞在移植部位的生长、分化,并通过检测局部CTGF、CD34的表达及Masson染色的情况比较瘢痕愈合情况.结果 移植后的干细胞在移植部位能很好的生长,并可向平滑肌细胞方向分化.在一定程度上可以降低局部的胶原纤维沉积,减轻瘢痕愈合.实验组CTGF的表达及Masson染色分别为0.0706

  20. Optimization of the method for isolating and culturing rat mesenchymal stem cells%大鼠骨髓间充质干细胞的分离与培养方法优化

    Institute of Scientific and Technical Information of China (English)

    祝旭龙; 颜谭; 姚维杰; 王永恒; 程冲; 向俊西; 吕毅; 高庆东; 李建辉

    2014-01-01

    目的:建立一个分离、培养、纯化大鼠的骨髓间充质干细胞的方法。方法使用贴壁法分离大鼠的骨髓间充质干细胞,通过培养初期的频繁换液和传代过程中减少胰蛋白酶的暴露时间,达到骨髓间充质干细胞的纯化。流式细胞仪检测细胞表面标志物,MTT法检测细胞生长状况,暴露于不同的诱导培养基使细胞成骨、成脂,并向肝胆系分化。结果分离所得的细胞阳性表达CD29、CD44、CD90,而对于造血细胞标志物CD45呈阴性。在成骨、成脂培养基的诱导下,细胞ALP、茜素红、油红O染色均呈阳性,并发现传至10代的细胞仍保持分化功能。将骨髓间充质干细胞贯续暴露于不同的细胞因子和激素,得到了表达肝细胞、胆管上皮细胞特异标志的子代细胞。结论通过改良后的方法可以高效、简洁地分离出形态均一的大鼠骨髓间充质干细胞,并能够向多胚层的细胞分化。%Objective To optimize the protocols for isolation and culture of mesenchymal stem cells from rat bone marrow (BMSCs). Methods BMSCs were isolated by adherence to plastic with frequent medium change and reduced trypsinization time. The cell growth curves were drawn and the surface markers of BMSCs were detected by flow cytometry. The cells were induced to differentiate into osteogenic, adipogenic, hepatic and cholic lineages. Results The cells isolated using this method were positive for CD29, CD44, and CD90 and negative for the hematopoietic surface markers CD45. The osteogenic and adipogenic differentiation of the BMSCs was verified by alkaline phosphatase staining, Alizarin red staining and Oil red staining. The cell subcultures up to passage 10 maintained capacities of differentiation into osteogenic and adipogenic lineages. The BMSCs induced with sequential addition of growth factors, cytokines and hormones differentiated into cells expressing hepatocyte-and cholangiocyte

  1. Experimental study of rat mesenchymal stem cells transfected with Sonic Hedgehog gene%Shh基因转染大鼠骨髓间充质干细胞的可行性

    Institute of Scientific and Technical Information of China (English)

    吴晓明; 唐滔; 杨进福; 谭志平; 张伟

    2012-01-01

    Objective To evaluate the feasibility of genetic modification of mesenchymal stem cells (MSC) with Sonic Hedgehog(Shh) gene.Methods The pcDNA3.l-Shh eukaryotic expression plasmid was constructed and its correctness evaluated by the restriction enzyme analysis and sequencing. MSC were isolated from Wistar rats by density gradient centrifugation and purified,transfected with pcDNA3.1-Shh,blank plasmid pcDNA3.1 ( - ) or pmaxGFP respectively by NucleofectorTM.The protein expression of Shh in MSC was detected by Western blot after 48 hours.Results Correct construction of pcDNA3.1-Shh was identified by the methods of restriction enzyme analysis and nucleotide sequence determination. The expression of green fluorescent protein (GFP) could be observed by fluorescence microscopy after 48 hours.The expression of Shh gene was detected by Western blot.But the MSC transfected with empty plasmid expression was not detected.Conclusions Recombinant Eukaryotic expression plasmid pcDNA3.1-Shh is successfully detected in rat MSC.It may provide experimental rationales for the future gene therapy.%目的 探讨Sonic Hedgehog(Shh)基因有效转染大鼠骨髓间充质干细胞(BMSC)的可行性.方法 构建pcDNA3.1-Shh真核表达质粒,酶切和测序鉴定.密度梯度离心-贴壁培养法获取Wistar大鼠BMSC,电击穿孔法将pcDNA3.1-Shh、pcDNA3.1(一)空质粒和pmaxGFP报告质粒转染进BMSC,48 h后Western印迹法检测Shh基因的表达情况.结果 酶切和测序结果证实pcDNA3.1-Shh正确性.转染48 h后,荧光显微镜可观察到绿色荧光蛋白(GFP)的表达,有40%以上的细胞发出绿色荧光.Western印迹法检测证实有Shh基因表达,而转染pcDNA3.1(一)空质粒的BBMSC未检测到表达.结论 重组真核表达质粒pcDNA3.1-Shh转染大鼠BMSC后能有效表达Shh基因,为进一步Shh基因与细胞联合治疗大鼠心肌缺血模型提供了实验依据.

  2. 小檗碱对大鼠骨髓间质干细胞成脂分化的抑制作用%Inhibitory effect of berberine on differentiation of rat bone marrow mesenchymal stem cells to adipocytes

    Institute of Scientific and Technical Information of China (English)

    徐道华; 杨玮; 周晨慧; 刘钰瑜; 许碧连

    2011-01-01

    OBJECTIVE To investigate the effect of berberine on differentiation of rat bone marrow mesenchymal stem cells (MSCs) to adipocytes and its mechanism. METHODS Rat MSCs were isolated and cultured, adipocytic differentiation was induced with adipogenesis-inducing medium (AIM). Cells were assigned into 6 groups:normal control, AIM group, AIM+berberine 0.1, 0.3, 1 and 3 μmol·L-1 groups, respectively. Morphology characteristics of mesenchymal stem cells were observed under an inverted microscope and adipocyte levels were analyzed by oil O staining. Alkaline phosphatase (ALP) activity was detected using p-nitrophenyl phosphate as a substrate. The cell survival was determined by MTT assay. Expressions of peroxisome proliferator activated receptor γ (PPARγ), fatty acid binding protein (aP2) and CCAAT enhancer-binding protein α (C/EBPα) mRNA were detected by semiquantitative RT-PCR. RESULTS Compared with normal control group, MSCs adipogenic differentiation, PPARγ, aP2 and C/EBPα mRNA expression significantly increased in AIM group (P<0.01), ALP activity in AIM group significantly decreased (P<0.01). Compared with AIM group, berberine inhibited MSCs adipogenic differentiation (P<0.01) and berberine 0.1, 0.3, 1 and 3 μmol·L-1 increased ALP activity by 26%, 54%, 81% and 122%, respectively. Berberine 3 μmol·L-1 significantly downregulated PPARγ expression (0.91±0.10 vs 1.34±0.06) (P<0.01), aP2 (1.05±0.10 vs 1.53±0.09) (P<0.01) and C/EBPα mRNA (1.24±0.06 vs 1.54±0.09) (P<0.01). Berberine had no effect on proliferation of MSCs. CONCLUSION Berberine inhibits differentiation of MSCs into adipocytes, which might be closely related to the downregulation of PPARγ, aP2 and C/EBPα mRNA.%目的 探讨小檗碱对大鼠骨髓间质干细胞成脂分化的影响及其机制.方法 经分离纯化的大鼠骨髓间质干细胞,分为正常对照组,成脂分化诱导液(AIM)模型组及AIM+小檗碱0.1,0.3,1和3 μmol·L-1组.倒置显微镜下观察细胞的形态特

  3. An in vivo tracing study on bone marrow mesenchymal stem cells in rats exposed to silica dust%骨髓间充质干细胞在染矽尘大鼠体内示踪研究

    Institute of Scientific and Technical Information of China (English)

    周永梅; 黄明; 燕玲; 曾子芳; 吴奇峰; 梁伟辉; 曾小菁

    2015-01-01

    Objective To investigate the distribution of bone marrow mesenchymal stem cell ( BMMSC) in rats exposed to silica dust.Methods BMMSCs were isolated from 7 specific pathogen free healthy male SD rats through bone marrow adherent method and transfected using lentiviral vector-enhanced green fluorescence protein ( LV-eGFP ) .Then Trypan blue staining and CCK-8 were applied to assay the cellular survival rate and cell proliferation activity before and after transfection, while 8 rats of the same kind were randomly divided into control group and silicotic group, with 4 rats in each group.Silicotic group was injected intratracheally with 1.0 mL silica suspension ( mass concentration was 40 g/L) while control group was given 1.0 mL physiological sodium chloride solution.The transfected BMMSCs were intravenously injected into the caudal veins of rats of the two groups.The lung, liver, spleen, heart, kidney and brain were taken and observed under the fluorescence microscopy by frozen section 1, 2, 3 and 4 weeks respectively after transplantation.The fluorescence distribution and of BMMSCs transplanted with LV-eGFP were detected, the fluorescence intensity of every organ was calculated by Image-pro plus 6.0.Results When the multiplicity of infection was 50, the transfection efficiency of BMMSCs was 80.00%, the green fluorescence expression was powerful and sustained, and there was not any change found in morphological characteristics of BMMSC after transfection.The respective cellular survival rates had no statistical significant difference between transfected and untransfected BMMSCs with LV-eGFP [ ( 97.67 ±0.60 )% vs (98.03 ±0.56 )%, P >0.05 ] .Cell proliferation activity had no statistical significant difference in main effects of transfection treatment (P >0.05).In the first week after transplantation of BMMSCs transfected with LV-eGFP, the fluorescence expression was observed in tissue sections of each organ in rats of two groups, the stronger fluorescence was

  4. In vivo regulation of erythropoiesis by chemically inducible dimerization of the erythropoietin receptor intracellular domain.

    Directory of Open Access Journals (Sweden)

    Norio Suzuki

    Full Text Available Erythropoietin (Epo and its receptor (EpoR are required for the regulation of erythropoiesis. Epo binds to the EpoR homodimer on the surface of erythroid progenitors and erythroblasts, and positions the intracellular domains of the homodimer to be in close proximity with each other. This conformational change is sufficient for the initiation of Epo-EpoR signal transduction. Here, we established a system of chemically regulated erythropoiesis in transgenic mice expressing a modified EpoR intracellular domain (amino acids 247-406 in which dimerization is induced using a specific compound (chemical inducer of dimerization, CID. Erythropoiesis is reversibly induced by oral administration of the CID to the transgenic mice. Because transgene expression is limited to hematopoietic cells by the Gata1 gene regulatory region, the effect of the CID is limited to erythropoiesis without adverse effects. Additionally, we show that the 160 amino acid sequence is the minimal essential domain of EpoR for intracellular signaling of chemically inducible erythropoiesis in vivo. We propose that the CID-dependent dimerization system combined with the EpoR intracellular domain and the Gata1 gene regulatory region generates a novel peroral strategy for the treatment of anemia.

  5. Observation of rat bone marrow mesenchymal stem cells after subretinally transplanted into rat retina%大鼠骨髓间充质干细胞视网膜下移植观察

    Institute of Scientific and Technical Information of China (English)

    张杰; 单清; 马萍; 姜严明; 陈鹏; 钱焕文; 裴雪涛

    2003-01-01

    目的鉴定骨髓间充质干细胞(mesenchymal stem cells,MSCs)在体外不诱导的条件下视网膜下移植后的定位.方法体外培养雄性大鼠MSCs,直接作为细胞供体视网膜下移植于成年雌性大鼠视网膜下,4周后将动物处死,取眼球做石蜡切片,用Y染色体鉴定;为做进一步验证,将MSCs用重组腺相关病毒AAV-gfp感染后移植,分别于4、8周取动物眼球作冰冻切片,于荧光显微镜下做绿色荧光蛋白(green fluorescence protein, GFP)表达观察.结果培养的MSCs集落生长迅速,均一性好;Y染色体原位杂交鉴定表明来源于MSCs的阳性细胞融合入了原来的视网膜结构,在光学显微镜下可分布于视锥、视杆细胞层、双极细胞层及节细胞层;在荧光显微镜下可见GFP标记的阳性细胞存在,分布于视网膜色素上皮层、视锥、视杆细胞层、双极细胞层及节细胞层,细胞形态与结构同周围的视网膜相似;2种标记方法检测到的视网膜结构完整,未见到玫瑰花结样结构.结论 MSCs可在视网膜下移植后4周与原视网膜结构相融合,2种方法检测到的阳性细胞分布于视网膜色素上皮层、视细胞层、双极细胞层及节细胞层.

  6. Transplantation of autologous bone marrow-derived mesenchymal stem cells for traumatic brain injury

    Institute of Scientific and Technical Information of China (English)

    Jindou Jiang; Xingyao Bu; Meng Liu; Peixun Cheng

    2012-01-01

    Results from the present study demonstrated that transplantation of autologous bone marrow-derived mesenchymal stem cells into the lesion site in rat brain significantly ameliorated brain tissue pathological changes and brain edema, attenuated glial cell proliferation, and increased brain-derived neurotrophic factor expression. In addition, the number of cells double-labeled for 5-bromodeoxyuridine/glial fibrillary acidic protein and cells expressing nestin increased. Finally, blood vessels were newly generated, and the rats exhibited improved motor and cognitive functions. These results suggested that transplantation of autologous bone marrow-derived mesenchymal stem cells promoted brain remodeling and improved neurological functions following traumatic brain injury.

  7. Serum from liver injury rats induces differentiation of umbilical cord mesenchymal stem cells into hepatocyte-like cells%肝损伤大鼠血清诱导培养脐带间充质干细胞向肝样细胞的分化

    Institute of Scientific and Technical Information of China (English)

    钟艳; 唐晓鹏

    2015-01-01

    背景:脐带间充质干细胞来源于新生个体脐带组织,来源广泛,无社会伦理学因素制约,有望代替骨髓间充质干细胞成为细胞移植和再生医学的理想种子细胞。目的:探讨肝衰竭大鼠血清对人脐带间充质干细胞肝向诱导分化作用,为临床上利用脐带间充质干细胞治疗终末期肝病提供实验依据。方法:腹腔注射10%四氯化碳溶液建立急性肝损伤大鼠模型,对照组腹腔注射等剂量的大豆油,48 h后腹主动脉取血离心分离血清。取第3代人脐带间充质干细胞,分别用体积分数为20%肝损伤大鼠血清和体积分数为20%胎牛血清诱导培养,观察诱导前后人脐带间充质干细胞形态学改变,检测培养液上清甲胎蛋白、白蛋白水平。结果与结论:肝损伤大鼠血清培养第1天细胞形态变化不大,呈梭形,仍呈编织状或漩涡状生长,第2天细胞呈短梭形,第3天细胞呈类圆形,第4天呈圆形,并有极少量细胞漂浮。肝损伤大鼠血清培养人脐带间充质干细胞4 d,其上清液白蛋白水平较诱导前和对照组均有升高趋势(P <0.001),甲胎蛋白水平无明显变化。结果表明肝损伤大鼠血清可使脐带间充质干细胞向肝样细胞分化。%BACKGROUND:Umbilical cord mesenchymal stem cels are from the umbilical cord of newly born individuals and have no ethical issues, and therefore are promising candidates for seeded cels as a substitute for cel transplantation and regenerative medicine. OBJECTIVE:To investigate the effects of serum from liver injury rats on induced differentiation of human umbilical cord mesenchymal stem cels into hepatocyte-like cels and provide experimental evidence for use of human umbilical cord mesenchymal stem cels in the treatment of patients with end-stage liver disease in the clinic. METHODS: Rat models of acute liver injury were established by intraperitoneal injection of 10% carbon

  8. Human telomerase reverse transcriptase transfection of bone marrow mesenchymal stem cells in the treatment of diabetic rats%人端粒酶反转录酶转染的骨髓间充质干细胞移植治疗大鼠糖尿病

    Institute of Scientific and Technical Information of China (English)

    赵海莲

    2015-01-01

    BACKGROUND:Human telomerase reverse transcriptase (hTERT) is the first choice for regulating the proliferation and directional differentiation, with multiple biological effects. OBJECTIVE:To observe the therapeutic effect of hTERT-transfected bone marrow mesenchymal stem cels transplantation in diabetic rats. METHODS: Bone marrow mesenchymal stem cels from Sprague-Dawley rats were culturedin vitro and transfected with retrovirus PLXSN carrying hTERT. RT-PCR was used to detect the hTERT expression in the bone marrow mesenchymal stem cels before and after transfection. Sixty male Sprague-Dawley rats were selected and equaly randomized into four groups: normal control group, transfection group, cel transplantation group, and model group. In the latter three groups, rats were injected with 45 mg/kg chain urea to establish diabetes models, and then injectedvia the tail vein with 0.2 mL hTERT-transfected bone marrow mesenchymal stem cels, 0.2 mL bone marrow mesenchymal stem cels, and 0.2 mL normal saline, respectively. RESULTS AND CONCLUSION:At 48 hours after hTERT transfection, the expression of hTERT mRNA was detected in the bone marrow mesenchymal stem cels, and mainly concentrated in the nuclei. At 14 days after transfection, the fasting glucose level in the model group was higher than that in the normal control group (P 0.05). These findings indicate that the transplantation of hTERT-transfected bone marrow mesenchymal stem cels is effective in the treatment of diabetic rats.%背景:人端粒酶反转录酶是调控增殖及定向分化的首选生长因子之一,具有多重生物学效应。目的:观察人端粒酶反转录酶表达的骨髓间充质干细胞移植治疗大鼠糖尿病的效果。方法:体外培养 SD 大鼠骨髓间充质干细胞,经反转录病毒 PLXSN 为载体介导人端粒酶反转录酶基因转染骨髓间充质干细胞,在转染前后用RT-PCR检测骨髓间充质干细胞人端粒酶反转录酶基因的表达。60

  9. 大鼠骨髓间充质干细胞成软骨过程中微小RNA的表达%The expression of miRNAs of rat bone marrow mesenchymal stem cells during chondrogenic differentiation

    Institute of Scientific and Technical Information of China (English)

    陈松; 符培亮; 吴海山; 许震宇

    2014-01-01

    Objective To explore the expression of miRNAs of rat bone marrow mesenchymal stem cells (BMSCs) during chondrogenic differentiation induced by Transforming growth factor-β3 (TGF-β3),and study the effect of over-expression of miR-90a on the differentiation of BMSCs toward chondrocytes.Methods The rat BMSCs were isolated and cultured invitro and were divided into two groups:treated with and without TGF-β3.The express of miRNAs were analyzed by microarray gene chip at 7,14,21 d,and the results were verified by real-time reverse transcriptase-polymerase chain reaction (RT-qPCR).Meanwhile the BMSCs treated with TGF-β3 were also divided into two groups,transfected with miR-90a mimics or mimics-NC,to explore the effect of over-expression of miR-90a on the chondrogenesis of BMSCs.Results The BMSCs were isolated from the rat marrow successful.Microarray gene chip screening resulted in 16 miRNAs whose expression differences were more than 2 folds compared with the controls.Of them,9 were up-regulated and 7 down-regulated.At the same time,the miR-340,miR-140,miR-21 and miR-132 were determined by RT-qPCR,and the results showed that RT-qPCR and microarray gene chip results are consistent.The group transfected with miR-90a mimics express fewer glycosaminoglycans (GAG) than that of transfected with mimics-NC.Conclusion Many miRNAs were involved in the process of BMSCs differentiation into chondrocytes,and over-expression of miR-90a could inhibit the process.%目的 探讨转化生长因子-β3(TGF-β3)诱导骨髓间充质干细胞(BMSCs)向软骨分化过程中微小RNA(miRNAs)的表达,并观察过表达miR-90a对BMSCs成软骨分化的影响.方法 体外分离培养大鼠BMSCs,取第3代BMSCs,在全培养基基础上将其分成两组:实验组加入TGF-β3诱导成软骨分化,对照组不加TGF-β3.在诱导BMSCs向软骨分化第7、14、21天,采用微阵列基因芯片技术分析分化过程中miRNAs的表达,并采用实时定量逆转录聚合酶链反应(RT-qPCR

  10. Influence of neuroplasticity of rat cerebral hemorrhage by bone marrow mesenchymal stem cells transplantation%骨髓间充质干细胞移植对大鼠脑出血后神经可塑性的影响

    Institute of Scientific and Technical Information of China (English)

    单泓; 李建斌; 刘敏; 戚正; 王姣杰; 韩小改; 梁会涛

    2013-01-01

    目的:观察骨髓间充质干细胞(BMSCs)移植对大鼠脑出血后神经可塑性的影响。方法利用脑立体定位仪制作大鼠脑出血模型;将120只大鼠随机分成BMSCs移植组、模型组,每组各60只;利用透射电镜观察血肿周围神经组织超微结构及神经突触的变化并应用Simple-PCI图像分析系统,选定测量参数计算突触数量,突触界面曲率,突触后致密区宽度及突触间隙;利用免疫组化技术检测大鼠血肿周围神经组织中神经突触相关蛋白Shank1、Nestin的表达;利用Berderson评分标准对大鼠进行神经功能评价。结果透射电镜观察,BMSCs移植组脑出血灶周围可见大量新生神经元及神经胶质细胞,神经突触数量为16.27±2.14,神经突触界面曲率为1.57±0.04,突触后致密区为68.32±10.54,突触间隙为14.65±1.58,与模型组比较差异有统计学意义(P<0.01)。BMSCs移植组及模型组Shank1表达分别为:75.82±10.65、14.33±1.14,两者比较差异有统计学意义(P<0.01);BMSCs移植组及模型组Nestin阳性细胞表达分别为:68.87±7.46、12.64±0.07,两者比较差异有统计学意义(P<0.01)。BMSCs移植组神经功能评分明显降低,与模型组比较有统计学意义(P<0.05)。结论 BMSCs 可促进大鼠脑出血后神经组织修复,增强神经可塑性,促进神经功能恢复。%Objective To observe the influence of neuroplasticity of hemorrhage in rats after transplantation of marrow mesenchymal stem cell. Methods The models were made by brain solid positioner. Rats were randomly divided into BMSCs group and control group.Observe the ultrastructure of nervous tissue and the change of neurosynaptic near hematoma by transmission electron microscope. Study the neurosynaptic quantificationally. The expression of Shank1and Nestin proteins which were related with synapse were detected by using immunohistochemistry. Evaluate the neural function

  11. 大鼠脂肪间充质干细胞移植治疗急性放射病的安全性%Safety of rat adipose-derived mesenchymal stem cell transplantation in the treatment of acute radiation sickness

    Institute of Scientific and Technical Information of China (English)

    许文黎; 张伟; 秦秀军; 岳娟; 李曙芳; 王永丽; 闻建华

    2013-01-01

    目的:探讨急性放射性大鼠经静脉输注异体脂肪间充质干细胞(adipose-derived mesenchymal stem cells,ADSCs)后的安全性。方法:SPF级雄性SD大鼠45只,随机分为空白对照组,照射对照组和照射移植组,每组15只。体外分离培养大鼠脂肪间充60质干细胞,照射对照组与照射移植组大鼠经 Co治疗机一次性7 Gy剂量全身照射后,照射移植组大鼠按10 mL/kg尾静脉注射脂肪间6充质干细胞,细胞浓度为1×10/mL;空白对照组和照射对照组注射等体积的无菌生理盐水。分别于照射后16和34 d进行血液学、血液生化学和组织病理学检查。结果:移植组大鼠体质量、血常规、血液生化与照射对照组相比差异无统计学意义(P>0.05);移植组大鼠脏器病理学观察与照射组、空白对照组相比无明显差别;未见移植组大鼠出现急慢性移植物抗宿主病反应。结论:急性放射性大鼠异体静脉输注脂肪间充质干细胞是安全可行的,对受者无不良影响。%OBJECTIVE: To investigate the safety of intravenous injection of allogeneic adipose-derived mesenchymal stem cells(ADSCs) in rats after acute radiation. METHODS:A total of 45 SPF male SD rats were equally and randomly divided into three groups:blank control group,radiation control group and radiation grafting group. Rat ADSCs were isolated and cultured in vitro. Irradiated control group and irradiated grafting rats were received 7 Gy dose 6total body irradiation once. Rats in each group received injection via tail vein,with 10 mL/kg of 1×10 ADSCs as the irradiated grafting group and the equal volume physiological saline as blank control group and irradiated control group. The rats were observed for 34 days for their general reactions before they were sacrificed. Blood samples were collected for hematological studies at the 16th day and biochemical studies at the 34th day. Visceral organs were inspected and studied

  12. Chemically induced transition phenomena in polyurethanes as seen from generalized mode Grueneisen parameters

    Energy Technology Data Exchange (ETDEWEB)

    Mueller, U; Philipp, M; Bactavatchalou, R; Sanctuary, R; Baller, J; Zielinski, B; Krueger, J K [Laboratoire de Physique des Materiaux, Universite du Luxembourg, 162A, Avenue de la Faiencerie, L-1115 (Luxembourg); Possart, W; Alnot, P [Laboratoire Europeen de Recherche, Universitaire Saarland-Lorraine (Luxembourg)], E-mail: ulrich.mueller@uni.lu

    2008-05-21

    Many phenomenological properties of reactive polymers like polyurethanes increase or decrease continuously in the course of the curing process before saturating at the end of the chemical reaction. This holds true for instance for the mass density, the refractive index, the chemical turnover and the hypersonic properties. The reason for this monotone behaviour is that the chemical reaction behaves like a continuous succession of irreversible phase transitions. These transitions are superposed by the sol-gel transition and possibly by the chemically induced glass transition, with the drawback that the latter two highlighted transitions are often hidden by the underlying curing process. In this work we propose generalized mode Grueneisen parameters as an alternative probe for elucidating the polymerization process itself and the closely related transition phenomena. As a model system we use polyurethane composed of a diisocyanate and varying ratios of difunctional and trifunctional alcohols.

  13. High Fidelity Tape Transfer Printing Based On Chemically Induced Adhesive Strength Modulation

    Science.gov (United States)

    Sim, Kyoseung; Chen, Song; Li, Yuhang; Kammoun, Mejdi; Peng, Yun; Xu, Minwei; Gao, Yang; Song, Jizhou; Zhang, Yingchun; Ardebili, Haleh; Yu, Cunjiang

    2015-11-01

    Transfer printing, a two-step process (i.e. picking up and printing) for heterogeneous integration, has been widely exploited for the fabrication of functional electronics system. To ensure a reliable process, strong adhesion for picking up and weak or no adhesion for printing are required. However, it is challenging to meet the requirements of switchable stamp adhesion. Here we introduce a simple, high fidelity process, namely tape transfer printing(TTP), enabled by chemically induced dramatic modulation in tape adhesive strength. We describe the working mechanism of the adhesion modulation that governs this process and demonstrate the method by high fidelity tape transfer printing several types of materials and devices, including Si pellets arrays, photodetector arrays, and electromyography (EMG) sensors, from their preparation substrates to various alien substrates. High fidelity tape transfer printing of components onto curvilinear surfaces is also illustrated.

  14. Heuristic model of chemically induced electron spin polarization in two dimensions

    Science.gov (United States)

    Adrian, Frank J.

    2010-11-01

    A heuristic model of chemically induced electron spin polarization (CIDEP) that breaks the polarization mechanism into its component steps, with each step governed by an appropriate solution of the diffusion equation, is extended from a three to a two-dimensional system. The required solution of the 2D diffusion equation is provided by a relatively simple analytic approximation to the usual infinite series solution. The model yields the polarization and its time development for weak to strong singlet-triplet mixing in the radical pairs, whereas previous models are limited to very weak or very strong mixing. Its results agree with a variational solution of an integral equation of Monchick and are encouraging for observation of CIDEP in dimensionally restricted systems. The method also may be applicable to other diffusion-controlled, spin-dependent chemistry in spatially restricted environments.

  15. Histopathological image analysis of chemical-induced hepatocellular hypertrophy in mice.

    Science.gov (United States)

    Asaoka, Yoshiji; Togashi, Yuko; Mutsuga, Mayu; Imura, Naoko; Miyoshi, Tomoya; Miyamoto, Yohei

    2016-04-01

    Chemical-induced hepatocellular hypertrophy is frequently observed in rodents, and is mostly caused by the induction of phase I and phase II drug metabolic enzymes and peroxisomal lipid metabolic enzymes. Liver weight is a sensitive and commonly used marker for detecting hepatocellular hypertrophy, but is also increased by a number of other factors. Histopathological observations subjectively detect changes such as hepatocellular hypertrophy based on the size of a hepatocyte. Therefore, quantitative microscopic observations are required to evaluate histopathological alterations objectively. In the present study, we developed a novel quantitative method for an image analysis of hepatocellular hypertrophy using liver sections stained with hematoxylin and eosin, and demonstrated its usefulness for evaluating hepatocellular hypertrophy induced by phenobarbital (a phase I and phase II enzyme inducer) and clofibrate (a peroxisomal enzyme inducer) in mice. The algorithm of this imaging analysis was designed to recognize an individual hepatocyte through a combination of pixel-based and object-based analyses. Hepatocellular nuclei and the surrounding non-hepatocellular cells were recognized by the pixel-based analysis, while the areas of the recognized hepatocellular nuclei were then expanded until they ran against their expanding neighboring hepatocytes and surrounding non-hepatocellular cells by the object-based analysis. The expanded area of each hepatocellular nucleus was regarded as the size of an individual hepatocyte. The results of this imaging analysis showed that changes in the sizes of hepatocytes corresponded with histopathological observations in phenobarbital and clofibrate-treated mice, and revealed a correlation between hepatocyte size and liver weight. In conclusion, our novel image analysis method is very useful for quantitative evaluations of chemical-induced hepatocellular hypertrophy.

  16. 12 hours after cerebral ischemia is the optimal time for bone marrow mesenchymal stem cell transplantation

    Directory of Open Access Journals (Sweden)

    Seyed Mojtaba Hosseini

    2015-01-01

    Full Text Available Cell therapy using stem cell transplantation against cerebral ischemia has been reported. However, it remains controversial regarding the optimal time for cell transplantation and the transplantation route. Rat models of cerebral ischemia were established by occlusion of the middle cerebral artery. At 1, 12 hours, 1, 3, 5 and 7 days after cerebral ischemia, bone marrow mesenchymal stem cells were injected via the tail vein. At 28 days after cerebral ischemia, rat neurological function was evaluated using a 6-point grading scale and the pathological change of ischemic cerebral tissue was observed by hematoxylin-eosin staining. Under the fluorescence microscope, the migration of bone marrow mesenchymal stem cells was examined by PKH labeling. Caspase-3 activity was measured using spectrophotometry. The optimal neurological function recovery, lowest degree of ischemic cerebral damage, greatest number of bone marrow mesenchymal stem cells migrating to peri-ischemic area, and lowest caspase-3 activity in the ischemic cerebral tissue were observed in rats that underwent bone marrow mesenchymal stem cell transplantation at 12 hours after cerebral ischemia. These findings suggest that 12 hours after cerebral ischemia is the optimal time for tail vein injection of bone marrow mesenchymal stem cell transplantation against cerebral ischemia, and the strongest neuroprotective effect of this cell therapy appears at this time.

  17. 12 hours after cerebral ischemia is the optimal time for bone marrow mesenchymal stem cell transplantation

    Institute of Scientific and Technical Information of China (English)

    Seyed Mojtaba Hosseini; Mohammad Farahmandnia; Zahra Razi; Somayeh Delavarifar; Benafsheh Shakibajahromi

    2015-01-01

    Cell therapy using stem cell transplantation against cerebral ischemia has been reported. However, it remains controversial regarding the optimal time for cell transplantation and the transplantation route. Rat models of cerebral ischemia were established by occlusion of the middle cerebral artery. At 1, 12 hours, 1, 3, 5 and 7 days after cerebral ischemia, bone marrow mesenchymal stem cells were injected via the tail vein. At 28 days after cerebral ischemia, rat neurological function was evaluated using a 6-point grading scale and the pathological change of ischemic cerebral tissue was observed by hematoxylin-eosin staining. Under the lfuorescence microscope, the migration of bone marrow mesenchymal stem cells was examined by PKH labeling. Caspase-3 activity was measured using spectrophotometry. The optimal neurological function recovery, lowest degree of ischemic cerebral damage, greatest number of bone marrow mesenchymal stem cells migrating to peri-ischemic area, and lowest caspase-3 activity in the ischemic cerebral tissue were observed in rats that underwent bone marrow mesenchymal stem cell transplantation at 12 hours after cerebral ischemia. These ifndings suggest that 12 hours after cerebral ischemia is the optimal time for tail vein injection of bone marrow mesenchymal stem cell transplantation against cerebral ischemia, and the strongest neuroprotective effect of this cell therapy appears at this time.

  18. Danhong injection plus bone marrow mesenchymal stem cell transplantation for treatment of cerebral infarction in rats%丹红注射液联合骨髓间充质干细胞移植治疗大鼠脑梗死★

    Institute of Scientific and Technical Information of China (English)

    张鹏; 周国庆; 孙晶晶

    2013-01-01

      背景:单纯骨髓间充质干细胞移植修复受损脑组织的作用并不十分理想。目的:观察丹红注射液联合骨髓间充质干细胞移植治疗大鼠脑梗死的效果。方法:用线栓法制备大鼠大脑中动脉阻塞模型,随机分为3组,模型组尾静脉注射PBS、丹红注射液组尾静脉注射2 mL/kg丹红注射液、联合治疗组联合注射2 mL/kg丹红注射液+2.0×109 L-1的骨髓间充质干细胞悬液,连续5 d,1次/d。结果与结论:在骨髓间充质干细胞移植后2周,联合治疗组神经功能评分明显优于模型组及丹红注射液组(P <0.05);移植后3周联合治疗组大鼠脑梗死体积明显小于模型组和丹红注射液组(P <0.05);病理组织学观察也可见联合治疗组的组织损伤减轻程度大于丹红注射液组和模型组。结果可见丹红注射液联合骨髓间充质干细胞移植治疗大鼠脑梗死疗效显著,可以对脑细胞起到保护作用。%  BACKGROUND: Simple bone marrow mesenchymal stem cel transplantation in repair of damaged brain tissues does not exhibit ideal effect. OBJECTIVE: To investigate the effect of Danhong injection combined with bone marrow mesenchymal stem cel transplantation on the treatment of cerebral infarction in rats. METHODS: The models of middle cerebral artery occlusion were established by suture method and randomly divided into three groups. Model group received tail vein injection of PBS. Danhong injection group received tail vein injection of 2 mL/kg Danhong injection. Combination group received injection of 2 mL/kg Danhong injection + 2.0×109/L bone marrow mesenchymal stem cel suspension, for 5 consecutive days, once a day. RESULTS AND CONCLUSION: At 2 weeks fol owing bone marrow mesenchymal stem cel transplantation, the neurological function scores in the combination group were significantly better than those in the model group and Danhong injection group (P < 0.05). Cerebral infarct volume in

  19. Protective effects of traditional Chinese medicine on mesenchymal stem cells transplanted into the hippocampus of rats with post-traumatic stress disorder%中药干预对创伤性应激障碍大鼠海马区移植间充质干细胞的保护作用***☆

    Institute of Scientific and Technical Information of China (English)

    富文俊; 敖海清; 曾蕾; 徐志伟

    2013-01-01

    BACKGROUND:Transplantation of bone marrow mesenchymal stem cel s shows great potential in repair of hippocampal injury. However, low survival rate of transplanted stem cel s remains unsolved. OBJECTIVE:To summarize the protective effects of traditional Chinese medicine on bone marrow mesenchymal stem cel s transplanted in the hippocampus of rats with post-traumatic stress disorder. METHODS:The databases of Pubmed, and CNKI were retrieved for papers published from January 2000 to January 2012 with key words“mesenchymal stem cel s, transplantation, survival and post-traumatic stress disorder (PTSD), hippocampus”in English and Chinese. A total of 109 papers were retrieved. Among them, 23 papers were included in the final analysis. RESULTS AND CONCLUSION:Post-traumatic stress disorder is a complex disease influenced by biological, psychological, social and other factors and remains unsolved in treatment. Traditional Chinese medicine exhibits a unique advantage in this research field. Hippocampal damage caused by post-traumatic stress disorder can be repaired by transplantation of bone marrow mesenchymal stem cel s. Bone marrow mesenchymal stem cel transplantation combined with intervention of Xiaoyaosan, a traditional Chinese medicine, is expected to increase the survival rate of transplanted stem cel s.%  背景:骨髓间充质干细胞移植修复损伤海马具有较高的研究潜力,但是目前移植后干细胞的成活率较低这一问题尚未得到有效解决。目的:综述中药干预对创伤性应激障碍大鼠海马区移植间充质干细胞的保护作用。方法:应用计算机检索2000年1月至2012年1月PubMed数据库相关文章,检索词为“mesenchymal stem cel s,transplantation,survival,post-traumatic stress disorder (PTSD),hippocampus”,并限定文章语言种类为English,同时检索2000年1月至2012年1月CNKI数据库相关文章,检索词为“间充质干细胞,移植,存活率,创伤后

  20. Effects of ethology,TChE and Ach activities in rat model of Middle cerebral artery occlusion administrated by human um- bilical cord-derived Mesenchymal stem cells%脐带间充质干细胞对脑梗死大鼠行为学及胆碱能的影响

    Institute of Scientific and Technical Information of China (English)

    贾芙蓉; 张贯石; 王辉; 潘洪涛; 何欣; 张东; 丁冬梅

    2014-01-01

    Objective To observe the effects of intravenous administration of human umbilical cord-derived mesen-chymal stem cells(UC-MSCs)after middle cerebral artery occlusion(MCAO)in adult rats.Methods Adult male SD rats were administrated by MCAO.Functional outcome measurements using Beam walking Test(BWT)were performed at 24 hour of MCAO group and 10 days of UC-MSCs treatment group respectively.TTC staining ,TChE and Ach activity array were also used.Results There was significant neurological function improvement in rats treated with MSC com-paring with that of control groups(P <0.05),and also Ach and TChE activity (P <0.05).Conclusion Intravenous ad-ministration of UC-MSCs promotes the neurological function in rat MCAO model.UC-MSCs play an advantageous function on cholinergic neurotransmitters in the case of lesion.%目的:观察脑梗死大鼠经人脐带间充质干细胞(umbilical cord-derived mesenchymal stem cells,UC-MSCs)治疗后,行为学的恢复情况,2,3,5-氯化三苯基四氮唑(triphenyl tetrazolium chloride,TTC)染色以及胆碱能的变化。方法制备脑梗死大鼠大脑中动脉闭塞(Middle cerebral artery occlusion,MCAO)模型,将 UC-MSCs 注入脑梗死大鼠体内,对大鼠运动功能进行评分,比较 TTC 染色显示的脑梗死区域所占比例,测定脑组织和血浆中乙酰胆碱酯酶(True choline esterase,TChE)活性和乙酰胆碱(Acetylcholine,Ach)浓度。结果与模型组相比,UC-MSCs 组大鼠行为学恢复明显(P <0.05),UC-MSCs 组大鼠脑组织和血浆乙酰胆碱酯酶(TChE)和乙酰胆碱(Ach)活性显著增强(P<0.05)。结论UC-MSCs 经尾静脉移植可促进脑梗死后神经功能恢复,增强大鼠胆碱能系统活性。

  1. Transplantation of placenta-derived mesenchymal stem cell-induced neural stem cells to treat spinal cord injur y

    Institute of Scientific and Technical Information of China (English)

    Zhi Li; Wei Zhao; Wei Liu; Ye Zhou; Jingqiao Jia; Lifeng Yang

    2014-01-01

    Because of their strong proliferative capacity and multi-potency, placenta-derived mesenchymal stem cells have gained interest as a cell source in the ifeld of nerve damage repair. In the present study, human placenta-derived mesenchymal stem cells were induced to differentiate into neural stem cells, which were then transplanted into the spinal cord after local spinal cord injury in rats. The motor functional recovery and pathological changes in the injured spinal cord were observed for 3 successive weeks. The results showed that human placenta-derived mesenchymal stem cells can differentiate into neuron-like cells and that induced neural stem cells contribute to the resto-ration of injured spinal cord without causing transplant rejection. Thus, these cells promote the recovery of motor and sensory functions in a rat model of spinal cord injury. Therefore, human placenta-derived mesenchymal stem cells may be useful as seed cells during the repair of spinal cord injury.

  2. Role for Kruppel-like transcription factor 11 in mesenchymal cell function and fibrosis.

    Directory of Open Access Journals (Sweden)

    Angela Mathison

    Full Text Available Krüppel-like factor 11 (KLF11 and the highly homologous KLF10 proteins are transcription factors originating from duplication of the Drosophila melanogaster ancestor cabut. The function of these proteins in epithelial cells has been previously characterized. In the current study, we report a functional role for KLF11 in mesenchymal cells and in mesenchymal cell dysfunction, namely, fibrosis, and subsequently perform a detailed cellular, molecular, and in vivo characterization of this phenomenon. We find that, in cultured mesenchymal cells, enhanced expression of KLF11 results in activated extracellular matrix pathways, including collagen gene silencing and matrix metalloproteinases activation without changes in tissue inhibitors of metalloproteinases. Combined, reporter and chromatin immunoprecipitation assays demonstrate that KLF11 interacts directly with the collagen 1a2 (COL1A2 promoter in mesenchymal cells to repress its activity. Mechanistically, KLF11 regulates collagen gene expression through the heterochromatin protein 1 gene-silencing pathway as mutants defective for coupling to this epigenetic modifier lose the ability to repress COL1A2. Expression studies reveal decreased levels of KLF11 during liver fibrogenesis after chemically induced injury in vivo. Congruently, KLF11(-/- mice, which should be deficient in the hypothesized anti-fibrogenic brake imposed by this transcription factor, display an enhanced response to liver injury with increased collagen fibril deposition. Thus, KLFs expands the repertoire of transcription factors involved in the regulation of extracellular matrix proteins in mesenchymal cells and define a novel pathway that modulates the fibrogenic response during liver injury.

  3. 骨髓间充质干细胞移植对慢性脑缺血大鼠海马区Cdc42表达及认知功能的影响%Transplantation of bone marrow mesenchymal stem cells increasing Cdc42 protein expression in the hippocampus and improving cognitive function of rats with chronic cerebral ischemia

    Institute of Scientific and Technical Information of China (English)

    于晓云; 张博爱; 李俊敏; 刘宇; 李晓晓; 崔璨; 崔红卫

    2013-01-01

    Objective To explore the effects of bone marrow mesenchymal stem cells transplantation on cognitive function and expression of cell division cycle 42 GTP-binding protein in hippocampus of chronic cerebral hypoperfusion rats. Methods 72 SD rats were randomly divided into sham, 2VO model (permanent ligation of bilateral common carotid artery) and experimental groups (2VO model + bone marrow mesenchymal stem cells transplantation). Rats in every group were subdivided into 8, 10 and 12 weeks subsets. Morris water maze was used to select rats of modeled successfully and space memory ability. Expression of Cdc42 protein in hippocampus was measured by Western blotting and immunohistochemistry. Results Expression of Cdc42 in hippocampus was reduced, escape latency was extended with time extending in experimental and model groups (P < 0. 05). Compared with model group, the expression of Cdc42 in the hippocampus tissue was increased and escape latency was shortened ( P < 0. 05 ) in experimental group. Conclusions Transplantation of bone marrow mesenchymal stem cells can improve cognitive function of chronic cerebral hypoperfusion rats, which maybe due to increased expression of Cdc42 protein.%目的 观察骨髓间充质干细胞移植后慢性脑缺血大鼠海马区表达Cdc42表达的变化,探讨其对慢性脑缺血大鼠认知功能的影响.方法 SD大鼠72只,随机分为假手术组、永久性双侧颈总动脉结扎(2VO)模型组、实验组(2VO模型+干细胞移植),每组分8、10、12w3个时间点,每个时间点8只,用Morris水迷宫筛选出造模成功的大鼠并检测大鼠的空间记忆能力,用Western印迹和免疫组化检测大鼠海马区Cdc42的表达.结果 实验组和模型组大鼠海马区Cdc42的表达随时间延长逐渐降低(P<0.05),其逃避潜伏期逐渐延长(P<0.05);较之2VO模型组,同一时间点实验组Cdc42表达明显增高(P<0.05),逃避潜伏期显著缩短(P<0.05).结论 骨髓间充质干细胞移植可显

  4. 骨髓间充质干细胞对糖尿病视网膜病变大鼠血糖及视网膜功能影响%Study the differentiated features and function of mesenchymal stem cells in rat model with diabetic retinopathy

    Institute of Scientific and Technical Information of China (English)

    利焕廉; 周金文; 蔡晓华

    2014-01-01

    目的:探讨尾静脉注射骨髓间充质干细胞对糖尿病视网膜病变大鼠血糖及视网膜功能的影响。方法:48只健康大鼠随机选取10只为正常对照组,38只建立糖尿病视网膜病变大鼠模型后,20只采用尾静脉注射间充质干细胞,18只注射等量安慰剂。观察间充质干细胞在肝、脾、肾、胰腺和眼球的分布以及在视网膜的分化特点,监测注射后1、2、3、4wk血糖变化,观察血-视网膜屏障破坏修复情况。结果:BMSCS在肝、脾、肾、胰腺、肺等组织中均有分布,视网膜外核层可见到大量密集的BMSCs重叠存在。实验组大鼠的血糖水平均随干预时间延长逐渐下降,注射后第1 wk血糖差异即有统计学意义。EB渗漏量较基线明显减少。结论:BMSCs可降低糖尿病视网膜病变大鼠血糖,使血-视网膜屏障得到一定程度修复。%Objective:To discussion the influence of blood glucose in diabetic retinopathy and retinal function in rats when injected the bone marrow mesenchymal stem cells on tail vein. Methods:Selecting 10 from the 48 rats randomly to be as normal control group, 38 rats after the establishment of diabetic retinopathy lesions, 20 using tail vein injection of mesenchymal stem cells and 18 the same amount of a placebo injection. Charge distribution was observed between mesenchymal stem cells in the liver, spleen, kidney, pancreas, and the characteristics of the eye and the retina differentiation. Observing the blood-retinal barrier breakdown repair the situation after1,2,3,4 weeks’ injection of blood glucose monitoring. Results:BMSCS are distributed in the liver, spleen, kidney, pancreas, and a large number of overlapping dense BMSCs can be seen in outer nuclear layer of the retina. Blood glucose average experimental rats gradually decreased with prolonged intervention, 1 week after injection blood sugar difference that is statistically signiifcant. EB leakage was signiifcantly

  5. 不同方式移植骨髓间充质干细胞在染矽尘大鼠体内的归巢%In vivo homing of bone marrow mesenchymal stem cells transplanted through different ways in rats exposed to silica dust

    Institute of Scientific and Technical Information of China (English)

    黄明; 周永梅; 燕玲; 李斌; 吴奇峰; 梁伟辉

    2015-01-01

    背景:骨髓间充质干细胞植入染矽尘大鼠体内能归巢到受损肺部,但何种植入途径在体内的归巢效果更好尚不清楚。  目的:动态比较观察不同途径移植骨髓间充质干细胞在染矽尘大鼠体内的分布情况。  方法:全骨髓贴壁法分离、培养供体大鼠骨髓间充质干细胞,用携带增强型绿色荧光蛋白基因的慢病毒(Lv-eGFP)转染骨髓间充质干细胞。受体大鼠用气管内注入法染尘,再随机分为经静脉组和经气管组,将转染Lv-eGFP的骨髓间充质干细胞分别经静脉、气管途径注入大鼠体内,在移植后的第1,2,3,4周处死大鼠,取心、肝、脾、肺、肾、脑组织进行冰冻切片,荧光显微镜下观察荧光,图文分析软件计算荧光强度。  结果与结论:两组大鼠肺组织均可见强烈、分布广泛且持续的绿色荧光,尤以气管、血管周围明显;两组荧光强度均随时间延长呈轻度减低趋势,但两组每周的荧光强度差异无显著性意义(P>0.05)。两组大鼠其余各脏器早期也均可见荧光,其中肝、脾、心脏组织荧光强、分布广,肾、脑组织荧光相对较弱、分布较稀疏;随时间推移,各组织荧光均逐渐减弱,面积逐渐减少,到后期仅肝、脾组织可见到较弱、散在荧光分布,脑组织荧光几乎不可见。在同一时间点,两组除第1周脑组织荧光强度差异有显著性意义(P0.05)。提示骨髓间充质干细胞经静脉和气管两种途径植入染矽尘大鼠体内归巢至受损肺部的效率相当。%BACKGROUND:Bone marrow mesenchymal stem cel s transplanted into rats exposed to silica dust can home to the injured lung, but the homing effects via different ways are stil unclear. OBJECTIVE:To comparatively observe the distribution of bone marrow mesenchymal stem cel s transplanted via different ways into rats exposed to silica dust. METHODS:Bone marrow

  6. Influence of tail vein administration of bone marrow mesenchymal stem cells on expression of brain-derived neurotrophic factor and nerve growth factor after spinal cord injury in rats%骨髓间充质干细胞尾静脉移植脊髓损伤大鼠脑源性神经营养因子及神经生长因子的表达

    Institute of Scientific and Technical Information of China (English)

    董锋; 林建华; 吴朝阳

    2011-01-01

    BACKGROUND: Transplantation of bone marrow mesenchymal stem cells has an effect on the treatment of spinal cord injury, but the mechanism is not fully understood.OBJECTIVE: To explore the influence of intravenous administration of bone marrow mesenchymal stem cells on expression of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) after spinal cord injury in rats and to study possible mechanism of intravenous administration of bone marrow mesenchymal stem cells after spinal cord injury in adult rats.METHODS: Modified Allen method was applied to induce T 10 spinal cord injury to prepare rat models of paraplegia. There were 6 rats in sham operation group and 24 rats in injury group. Then, the injury group was randomly divided into transplantation group and control group. Rats in the sham operation and transplantation groups received tail vein transplantation of 1 mL bone marrow mesenchymal stem cells (1×106 cells), while those in the control group were treated with PBS.RESULTS AND CONCLUSION: After spinal cord injury in rats, the expression of BDNF and NGF was improved and advanced to increase via intravenous administration of bone marrow mesenchymal stem cells. Maybe it is one of the mechanisms of promoting restoration of injured spinal cord and improving motor function.%背景:骨髓间充质干细胞移植对脊髓损伤有治疗作用,但其机制尚不完全清楚.目的:应用免疫组织化学方法观察骨髓间充质干细胞静脉移植损伤脊髓局部脑源性神经营养因子及神经生长因子的表达,分析骨髓间充质干细胞移植治疗大鼠脊髓损伤的作用途径.方法:运用改良Allen 法制备T10 脊髓外伤性截瘫大鼠模型,假手术组6 只,脊髓损伤组24 只随机分为对照组和骨髓间充质干细胞移植组.骨髓间充质干细胞移植组、假手术组接受骨髓间充质干细胞单细胞悬液1 mL(1×106 cells)自大鼠尾静脉缓慢注射移植,对照组静脉注射PBS 1 mL.结果

  7. Bone marrow mesenchymal stem cells repair spinal cord ischemia/reperfusion injur y by promoting axonal growth and anti-autophagy

    Institute of Scientific and Technical Information of China (English)

    Fei Yin; Chunyang Meng; Rifeng Lu; Lei Li; Ying Zhang; Hao Chen; Yonggang Qin; Li Guo

    2014-01-01

    Bone marrow mesenchymal stem cells can differentiate into neurons and astrocytes after trans-plantation in the spinal cord of rats with ischemia/reperfusion injury. Although bone marrow mesenchymal stem cells are known to protect against spinal cord ischemia/reperfusion injury through anti-apoptotic effects, the precise mechanisms remain unclear. In the present study, bone marrow mesenchymal stem cells were cultured and proliferated, then transplanted into rats with ischemia/reperfusion injury via retro-orbital injection. Immunohistochemistry and immunolfuorescence with subsequent quantiifcation revealed that the expression of the axonal regeneration marker, growth associated protein-43, and the neuronal marker, microtubule-as-sociated protein 2, significantly increased in rats with bone marrow mesenchymal stem cell transplantation compared with those in rats with spinal cord ischemia/reperfusion injury. Fur-thermore, the expression of the autophagy marker, microtubule-associated protein light chain 3B, and Beclin 1, was signiifcantly reduced in rats with the bone marrow mesenchymal stem cell transplantation compared with those in rats with spinal cord ischemia/reperfusion injury. Western blot analysis showed that the expression of growth associated protein-43 and neuro-iflament-H increased but light chain 3B and Beclin 1 decreased in rats with the bone marrow mesenchymal stem cell transplantation. Our results therefore suggest that bone marrow mes-enchymal stem cell transplantation promotes neurite growth and regeneration and prevents autophagy. These responses may likely be mechanisms underlying the protective effect of bone marrow mesenchymal stem cells against spinal cord ischemia/reperfusion injury.

  8. 无细胞神经移植物复合骨髓间充质干细胞修复大鼠坐骨神经缺损%Repairing rat sciatic nerve defects with the compound of acellular nerve graft and bone marrow mesenchymal stem cells

    Institute of Scientific and Technical Information of China (English)

    张彩顺; 吕刚; 张基仁; 肖强

    2009-01-01

    BACKGROUND: Previous studies have successfully prepared the natural and biologically degraded acellular nerve graft and have proved the effect of promoting neural regeneration.OBJECTIVE: To construct tissue engineered artificial nerve with acellular nerve graft and bone marrow mesenchymal stem cells, and to observe the effect of promoting motor functional recovery and repairing rat sciatic nerve defects. DESIGN, TIME AND SETTING: Randomized control animal experiment was performed in the Medical TIssue Engineering Laboratory of the First Affiliated Hospital of Liaoning Medical University between June 2008 and February 2009. MATERIALS: Wistar adult healthy male rats weighing 180-200 g were used to prepare acellular nerve graft, while Wistar adult healthy male rats weighing 100-120 g were used to prepare bone marrow mesenchymal stem cells. Tissue engineered artificial nerve was produced with acellular nerve graft co-cultured with bone marrow mesenchymal stem cells. METHODS: Sixty Wistar adult healthy male rats weighing 180-200 g were induced sciatic nerve defect models, 15 mm long. SD rats were divided into three groups at random with 20 animals in each group. ①Experiment group: Rat sciatic nerve defects were bridged with tissue engineered artificial nerve. ②Blank control group: Rat sciatic nerve defects were bridged with tissue engineered nerve scaffold. ③Autologous nerve control group: Rat sciatic nerve defects were bridged with autologous nerve graft. MAIN OUTCOME MEASURES: At 12 weeks postoperation, the recovery of motor function was evaluated with gross observation, electrophysiology, histological observation and triceps surae wet weight.RESULTS: ①At 12 weeks postoperation, the toes at the operation side could separate and supported to the ground in the experiment group; there was no significant difference in the regenerated nerve conduction velocity between experimental group and autologous nerve graft group. ②At 12 weeks postoperation, histochemical

  9. 骨髓间充质干细胞对急性百草枯中毒肺损伤的保护作用%Protective effects of bone marrow mesenchymal stem cells on paraquat-induced acute lung injury in rats

    Institute of Scientific and Technical Information of China (English)

    黄杨; 尹文; 刘健; 余厚友; 王玉同; 郝璐

    2011-01-01

    目的:通过骨髓间充质干细胞(BMSCs)的植入,观察其对急性百草枯(paraqua)中毒肺损伤的保护效果.方法:54只雌性受体大鼠随机分4组:百草枯组、BMSCs治疗组、BMSCs对照组和正常对照组,将体外分离和培养的BMSCs通过尾静脉注入受体体内,观察14 d内各组大鼠的存活率、肺干湿比、血浆中IL-1β、TNF-α、MDA、SOD、GSH- PX的水平变化.结果:与百草枯组相比,BMSCs治疗组明显延长大鼠的生存期,降低肺干湿比(P < 0.05)和炎症介质IL-1β、TNF-α的含量(P < 0.01),降低血浆中MDA的水平(P < 0.05)和升高SOD、GSH- PX的水平(P < 0.01).结论: BMSCs对急性百草枯中毒肺损伤具有保护作用.%Objective To investigate the protective effects of bone marrow mesenchymal stem cells (BMSCs) on paraquat-induced acute lung injury in rats. Methods BMSCs were isolated from male SD rats and injected into recipient SD rats after cultured in vitro. 54 female recipient SD rats were divided randomly into four groups:PQ group, BMSCs treatment group, BMSCs control group and normal control group. The rats survival time and lung wet/dry weight ratios were calculated during 14 days after the injections in each group. At the same time, the levels of Interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) , malondialdehyde (MDA) ,superoxide dismutase (SOD) and glutathione peroxidase (GSH- PX) in plasma were all detected. ResultsCompared with the PQ group, the suvival time of recipient rats was prolonged and lung wet/dry weight ratios were down obviously (P < 0.05), the levels of IL-1β, TNF-α and MDA in plasma were decreased markedly,and the activities of SOD、GSH- PX in plasma were increased significantly in the BMSCs treatment group.Conclusion BMSCs have protective effects on paraquat-induced acute lung injury in rats.

  10. Conditioned Medium From Human Amniotic Mesenchymal Stromal Cells Limits Infarct Size and Enhances Angiogenesis

    NARCIS (Netherlands)

    Danieli, Patrizia; Malpasso, Giuseppe; Cluffreda, Maria Chiara; Cervio, Elisabetta; Calvillo, Laura; Copes, Francesco; Pisano, Federica; Mura, Manuela; Kleijn, Lennaert; de Boer, Rudolf A.; Viarengo, Gianluca; Rosti, Vittorio; Spinillo, Arsenio; Roccio, Marianna; Gnecchi, Massimiliano

    2015-01-01

    The paracrine properties of human amniotic membrane-derived mesenchymal stromal cells (hAMCs) have not been fully elucidated. The goal of the present study was to elucidate whether hAMCs can exert beneficial paracrine effects on infarcted rat hearts, in particular through cardioprotection and angiog

  11. 骨髓间充质干细胞复合聚乳酸-羟基乙酸材料治疗大鼠神经损伤的实验研究%Bone Marrow Mesenchymal Stem Cells Combined with Polylactic Glycolic Acid Scaffold for Rat Neural Injury

    Institute of Scientific and Technical Information of China (English)

    符厚圣; 周兴; 桂有富; 郑煜; 潘建刚

    2011-01-01

    Objectives To study separately the effect of bone marrow mesenchymal stem cells (BMSCs) and differentiated neural - like cells combined with polylactic glycolic acid ( PLGA ) scaffolds, both composite which restore to SD rat nerve injury. Methods BMSCs were cultured by differential adherence,and composited PLGA,via scanning electron microscope to investigate the information of BMSCs'adhesion, proliferation and induction into neural - like cells in the scaffolds. Bold scaffold, BMSCs - PLGA complex, and neural - like cells - PLGA complex were separately transplanted into SD rat models, with the injury for right sciatic nerve, and obsvered SD rat models during 60 days after operation. Results BMSCs and neural - like cells can be exactly adhere to, proliferate and grow on the PLGA;BMSCs and neural -like cells combined with PLGA, which can restore to nerve injury. Conclusions BMSCs and differentiated neural - like cells combined with polylactic glycolic acid (PLGA) scaffolds,which can be used as a method of tissue engineering,to treat neurogenic bladder caused by spinal cord injury and advanced diabetes.%目的 研究骨髓间充质干细胞(Bone Marrow Mesenchymal Stem Cells,BMSCs)及其分化的神经样细胞分别与聚乳酸-羟基乙酸(polylactic glycolic acid,PLGA)支架材料复合修复大鼠神经损伤的效果.方法 将BMSCs复合PIGA培养,通过扫描电镜观察BMSCs在PLGA上的黏附、增殖并诱导分化成神经样细胞的情况;然后分别将单纯的PLGA、BMSCs-PLGA复合物以及神经样细胞-PLGA复合物分别移植入3组右侧坐骨神经损伤的SD大鼠模型中,术后观察三组动物60 d.结果 BMSCs及诱导分化生成的神经样细胞能在PLGA良好的黏附、增殖和生长;复合PLGA移植后能修复损伤的神经.结论 BMSCs及神经样细胞复合PLGA移植可作为组织工程治疗脊髓损伤及糖尿病晚期神经源性膀胱的一种方法.

  12. Isolation, culture and identification of nucleus pulposus-derived mesenchymal stem cells from adult rats in vitro%大鼠椎间盘髓核来源间充质干细胞的体外分离及培养鉴定

    Institute of Scientific and Technical Information of China (English)

    芮云峰; 王善正; 谢鑫荟; 孙明辉; 林禹丞; 李刚; 王宸

    2013-01-01

    背景:目前椎间盘髓核组织的细胞组成和特性仍未阐明。  目的:旨在建立大鼠椎间盘髓核来源间充质干细胞的体外培养体系,并对其体外多项分化潜能进行鉴定。  方法:体外培养SD大鼠盘髓核来源间充质干细胞,取第3代细胞进行三系诱导分化,将成骨、成脂和成软骨分化作为实验组,基础细胞培养作为对照组。  结果与结论:低密度培养获得的髓核来源细胞早期可形成葵花样细胞集落,克隆样生长。第3代后,细胞形态趋向均一,呈成纤维细胞样生长。成骨诱导28 d,实验组茜素红染色阳性,且 RunX2、osteopontin 及osteocalcin表达较对照组显著增高(P OBJECTIVE:To establish the in vitro culture system of rat nucleus pulposus-derived mesenchymal stem cells and to identify their multi-lineage differentiation potential. METHODS:Mesenchymal stem cells from the nucleus pulposus tissues of Sprague-Dawley rats were cultured in vitro. Then, cells at passage 3 were induced to differentiate into osteoblasts, adipocytes and chondrocytes as experimental group. cells cultured with basic culture medium served as controls. RESULTS AND CONCLUSION:cells isolated from rat nucleus pulposus could form the sunflower-like colonies and exhibit clone-like growth when they cultured at a low density. cells at passage 3 became homogeneous and exhibited fibroblast-like morphology. After 28 days of osteogenic induction, arizarin red positive signals were detected in the experimental group. The mRNA expressions of RunX2, osteopontin and osteocalcin were significantly increased in the experimental group, compared to the control group (P<0.05). After 21 days of adipogenic induction, oil red-O positive cells were detected in the experimental group. The mRNA expressions of C/EBPαand PPARγ2 were significantly increased in the experimental group, compared to the control group (P<0.05). After 21 days of chondrogenic

  13. Chemically induced graft copolymerization of 2-hydroxyethyl methacrylate onto polyurethane surface for improving blood compatibility

    Science.gov (United States)

    He, Chunli; Wang, Miao; Cai, Xianmei; Huang, Xiaobo; Li, Li; Zhu, Haomiao; Shen, Jian; Yuan, Jiang

    2011-11-01

    To improve hydrophilicity and blood compatibility properties of polyurethane (PU) film, we chemically induced graft copolymerization of 2-hydroxyethyl methacrylate (HEMA) onto the surface of polyurethane film using benzoyl peroxide as an initiator. The effects of grafting temperature, grafting time, monomer and initiator concentrations on the grafting yields were studied. The maximum grafting yield value was obtained 0.0275 g/cm2 for HEMA. Characterization of the films was carried out by attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR), water contact angle measurements. ATR-FTIR data showed that HEMA was successfully grafted onto the PU films surface. Water contact angle measurement demonstrated the grafted films possessed a relatively hydrophilic surface. The blood compatibility of the grafted films was preliminarily evaluated by a platelet-rich plasma adhesion test and hemolysis test. The results of platelet adhesion experiment showed that polyurethane grafted polymerization with monomer of 2-hydroxyethyl methacrylate had good blood compatibility featured by the low platelet adhesion. Hemolysis rate of the PU-g-PHEMA films was dramatically decreased than the ungrafted PU films. This kind of new biomaterials grafted with HEMA monomers might have a potential usage for biomedical applications.

  14. Dietary zinc deficiency predisposes mice to the development of preneoplastic lesions in chemically-induced hepatocarcinogenesis.

    Science.gov (United States)

    Romualdo, Guilherme Ribeiro; Goto, Renata Leme; Henrique Fernandes, Ana Angélica; Cogliati, Bruno; Barbisan, Luis Fernando

    2016-10-01

    Although there is a concomitance of zinc deficiency and high incidence/mortality for hepatocellular carcinoma in certain human populations, there are no experimental studies investigating the modifying effects of zinc on hepatocarcinogenesis. Thus, we evaluated whether dietary zinc deficiency or supplementation alter the development of hepatocellular preneoplastic lesions (PNL). Therefore, neonatal male Balb/C mice were submitted to a diethylnitrosamine/2-acetylaminefluorene-induced hepatocarcinogenesis model. Moreover, mice were fed adequate (35 mg/kg diet), deficient (3 mg/kg) or supplemented (180 mg/kg) zinc diets. Mice were euthanized at 12 (early time-point) or 24 weeks (late time-point) after introducing the diets. At the early time-point, zinc deficiency decreased Nrf2 protein expression and GSH levels while increased p65 and p53 protein expression and the number of PNL/area. At the late time-point, zinc deficiency also decreased GSH levels while increased liver genotoxicity, cell proliferation into PNL and PNL size. In contrast, zinc supplementation increased antioxidant defense at both time-points but not altered PNL development. Our findings are the first to suggest that zinc deficiency predisposes mice to the PNL development in chemically-induced hepatocarcinogenesis. The decrease of Nrf2/GSH pathway and increase of liver genotoxicity, as well as the increase of p65/cell proliferation, are potential mechanisms to this zinc deficiency-mediated effect.

  15. Combining machine learning, crowdsourcing and expert knowledge to detect chemical-induced diseases in text.

    Science.gov (United States)

    Bravo, Àlex; Li, Tong Shu; Su, Andrew I; Good, Benjamin M; Furlong, Laura I

    2016-01-01

    Drug toxicity is a major concern for both regulatory agencies and the pharmaceutical industry. In this context, text-mining methods for the identification of drug side effects from free text are key for the development of up-to-date knowledge sources on drug adverse reactions. We present a new system for identification of drug side effects from the literature that combines three approaches: machine learning, rule- and knowledge-based approaches. This system has been developed to address the Task 3.B of Biocreative V challenge (BC5) dealing with Chemical-induced Disease (CID) relations. The first two approaches focus on identifying relations at the sentence-level, while the knowledge-based approach is applied both at sentence and abstract levels. The machine learning method is based on the BeFree system using two corpora as training data: the annotated data provided by the CID task organizers and a new CID corpus developed by crowdsourcing. Different combinations of results from the three strategies were selected for each run of the challenge. In the final evaluation setting, the system achieved the highest Recall of the challenge (63%). By performing an error analysis, we identified the main causes of misclassifications and areas for improving of our system, and highlighted the need of consistent gold standard data sets for advancing the state of the art in text mining of drug side effects.Database URL: https://zenodo.org/record/29887?ln¼en#.VsL3yDLWR_V.

  16. Insights from advances in research of chemically induced experimental models of human inflammatory bowel disease

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Inflammatory bowel disease (IBD), the most important being Crohn's disease and ulcerative colitis, results from chronic dysregulation of the mucosal immune system in the gastrointestinal tract. Although the pathogenesis of IBD remains unclear, it is widely accepted that genetic, environmental, and immunological factors are involved. Recent studies suggest that intestinal epithelial defenses are important to prevent inflammation by protecting against microbial pathogens and oxidative stresses. To investigate the etiology of IBD, animal models of experimental colitis have been developed and are frequently used to evaluate new anti-inflammatory treatments for IBD. Several models of experimental colitis that demonstrate various pathophysiological aspects of the human disease have been described. In this manuscript, we review the characteristic features of IBD through a discussion of the various chemically induced experimental models of colitis (e.g. dextran sodium sulfate-, 2,4,6-trinitrobenzene sulfonic acid-, oxazolone-, acetic acid-, and indomethacin-induced models). We also summarize some regulatory and pathogenic factors demonstrated by these models that can, hopefully, be exploited to develop future therapeutic strategies against IBD.

  17. A mint purified extract protects human keratinocytes from short-term, chemically induced oxidative stress.

    Science.gov (United States)

    Berselli, Patrizia Valeria Rita; Zava, Stefania; Montorfano, Gigliola; Corsetto, Paola Antonia; Krzyzanowska, Justyna; Oleszek, Wieslaw; Berra, Bruno; Rizzo, Angela Maria

    2010-11-10

    Oxidative stress is strictly correlated to the pathogenesis of many diseases, and a diet rich in fruits and vegetables, or adequately integrated, is currently considered to be a protective and preventive factor. This study aimed to analyze the efficacy of a 1 h preincubation with the highest nontoxic dose of a characterized Mentha longifolia extract (80 μg/mL) in protecting human keratinocytes (NCTC2544) from chemically induced oxidative stress (500 μM H2O2 for 2, 16, and 24 h). As reference synthetic pure compounds rosmarinic acid (360.31 μg/mL), a major mint phenolic constituent, and resveratrol (31.95 mg/mL), a well-known antioxidant, were used. Cellular viability was significantly protected by mint, which limited protein and DNA damage, decreased lipid peroxidation, and preserved glutathione and superoxide dismutase activity in the shorter phases of oxidative stress induction, in extents comparable to or better than those of pure compounds. These data suggest that mint use as only a flavoring has to be revised, taking into consideration its enrichment in foodstuff and cosmetics.

  18. Proteomic Analyses of Cellular Events Mediating/Inhibiting Chemical-Induced Injury

    Science.gov (United States)

    2009-07-21

    Yan L, Wang X, Pei R, Van T, Zhao Y, Guo X: Cytotoxicity of carbon nanomaterials: single-wall nanotube , multi·wall nanotube , and fullerene. Environ...data and functional analyses. 14. ABSTRACT 15. SUBJECT TERMS 16. SECURITY CLASSIFICATION OF: 17. LIMITATION OF ABSTRACT 18. NUMBER OF...post-formalin exposure. rat lungs were removed and frozen for 2-DE. 2.1.2 Lung Function Test Lung function in replicate rats was characterized as

  19. Rats

    Directory of Open Access Journals (Sweden)

    Alexey Kondrashov

    2012-01-01

    Full Text Available We aimed to perform a chemical analysis of both Alibernet red wine and an alcohol-free Alibernet red wine extract (AWE and to investigate the effects of AWE on nitric oxide and reactive oxygen species production as well as blood pressure development in normotensive Wistar Kyoto (WKY and spontaneously hypertensive rats (SHRs. Total antioxidant capacity together with total phenolic and selected mineral content was measured in wine and AWE. Young 6-week-old male WKY and SHR were treated with AWE (24,2 mg/kg/day for 3 weeks. Total NOS and SOD activities, eNOS and SOD1 protein expressions, and superoxide production were determined in the tissues. Both antioxidant capacity and phenolic content were significantly higher in AWE compared to wine. The AWE increased NOS activity in the left ventricle, aorta, and kidney of SHR, while it did not change NOS activity in WKY rats. Similarly, increased SOD activity in the plasma and left ventricle was observed in SHR only. There were no changes in eNOS and SOD1 expressions. In conclusion, phenolics and minerals included in AWE may contribute directly to increased NOS and SOD activities of SHR. Nevertheless, 3 weeks of AWE treatment failed to affect blood pressure of SHR.

  20. Mammary phenotypic expression induced in epidermal cells by embryonic mammary mesenchyme.

    Science.gov (United States)

    Cunha, G R; Young, P; Christov, K; Guzman, R; Nandi, S; Talamantes, F; Thordarson, G

    1995-01-01

    The goal of this research was to establish methods for inducing mammary epithelial differentiation from nonmammary epithelium. For this purpose, mid-ventral or dorsal epidermis (skin epithelium; SKE) from 13-day rat or mouse embryos was associated with 13-day embryonic mouse mammary mesenchyme (mammary gland mesenchyme; MGM) (mouse MGM+rat or mouse SKE). The resultant MGM+SKE recombinants as well as controls (homotypic mouse mammary recombinants, homotypic mouse skin recombinants and mouse mammary mesenchyme by itself) were grafted under the renal capsule of syngeneic or athymic female nude mouse hosts. Most female hosts were induced to undergo lactogenesis by grafting an adult pituitary which elicited a state of hyperprolactinemia. Tissue recombinants of mouse MGM+rat or mouse SKE grown for 1 month in vivo formed a hair-bearing keratinized skin from which mammary ductal structures extended into the mesenchyme. The ducts were composed of columnar luminal epithelial cells as well as basal, actin-positive myoepithelial cells. When grown in pituitary-grafted hosts, the ductal epithelial cells expressed casein and alpha-lactalbumin as judged by immunocytochemistry. The expression of caseins in MGM+SKE recombinants was confirmed by Western blot. The epithelial cells in mouse MGM+rat SKE recombinants expressing milk proteins were shown to be rat cells while the surrounding connective tissue was composed of mouse cells based upon staining with Hoechst dye 33258. Using mammary-specific markers, these studies confirmed the earlier morphological studies of Propper and unequivocally demonstrated for the first time that embryonic mammary mesenchyme can induce morphological and functional mammary differentiation from nonmammary epithelium.

  1. Dorsal root ganglion neurons promote proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells

    Institute of Scientific and Technical Information of China (English)

    Pei-xun Zhang; Xiao-rui Jiang; Lei Wang; Fang-min Chen; Lin Xu; Fei Huang

    2015-01-01

    Preliminary animal experiments have conifrmed that sensory nerve ifbers promote osteoblast differentiation, but motor nerve ifbers have no promotion effect. Whether sensory neurons pro-mote the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells remains unclear. No results at the cellular level have been reported. In this study, dorsal root ganglion neurons (sensory neurons) from Sprague-Dawley fetal rats were co-cultured with bone marrow mesenchymal stem cells transfected with green lfuorescent protein 3 weeks after osteo-genic differentiationin vitro, while osteoblasts derived from bone marrow mesenchymal stem cells served as the control group. The rat dorsal root ganglion neurons promoted the prolifera-tion of bone marrow mesenchymal stem cell-derived osteoblasts at 3 and 5 days of co-culture, as observed by lfuorescence microscopy. The levels of mRNAs for osteogenic differentiation-re-lated factors (including alkaline phosphatase, osteocalcin, osteopontin and bone morphogenetic protein 2) in the co-culture group were higher than those in the control group, as detected by real-time quantitative PCR. Our ifndings indicate that dorsal root ganglion neurons promote the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells, which pro-vides a theoretical basis forin vitro experiments aimed at constructing tissue-engineered bone.

  2. Visual bone marrow mesenchymal stem cell transplantation in the repair of spinal cord injury

    Directory of Open Access Journals (Sweden)

    Rui-ping Zhang

    2015-01-01

    Full Text Available An important factor in improving functional recovery from spinal cord injury using stem cells is maximizing the number of transplanted cells at the lesion site. Here, we established a contusion model of spinal cord injury by dropping a weight onto the spinal cord at T 7-8 . Superparamagnetic iron oxide-labeled bone marrow mesenchymal stem cells were transplanted into the injured spinal cord via the subarachnoid space. An outer magnetic field was used to successfully guide the labeled cells to the lesion site. Prussian blue staining showed that more bone marrow mesenchymal stem cells reached the lesion site in these rats than in those without magnetic guidance or superparamagnetic iron oxide labeling, and immunofluorescence revealed a greater number of complete axons at the lesion site. Moreover, the Basso, Beattie and Bresnahan (BBB locomotor rating scale scores were the highest in rats with superparamagnetic labeling and magnetic guidance. Our data confirm that superparamagnetic iron oxide nanoparticles effectively label bone marrow mesenchymal stem cells and impart sufficient magnetism to respond to the external magnetic field guides. More importantly, superparamagnetic iron oxide-labeled bone marrow mesenchymal stem cells can be dynamically and non-invasively tracked in vivo using magnetic resonance imaging. Superparamagnetic iron oxide labeling of bone marrow mesenchymal stem cells coupled with magnetic guidance offers a promising avenue for the clinical treatment of spinal cord injury.

  3. visual bone marrow mesenchymal stem cell transplantation in the repair of spinal cord injury

    Institute of Scientific and Technical Information of China (English)

    Rui-ping Zhang; Cheng Xu; Yin Liu; Jian-ding Li; Jun Xie

    2015-01-01

    An important factor in improving functional recovery from spinal cord injury using stem cells is maximizing the number of transplanted cells at the lesion site. Here, we established a contusion model of spinal cord injury by dropping a weight onto the spinal cord at T7–8. Superparamagnet-ic iron oxide-labeled bone marrow mesenchymal stem cells were transplanted into the injured spinal cordvia the subarachnoid space. An outer magnetic ifeld was used to successfully guide the labeled cells to the lesion site. Prussian blue staining showed that more bone marrow mesen-chymal stem cells reached the lesion site in these rats than in those without magnetic guidance or superparamagnetic iron oxide labeling, and immunolfuorescence revealed a greater number of complete axons at the lesion site. Moreover, the Basso, Beattie and Bresnahan (BBB) locomotor rating scale scores were the highest in rats with superparamagnetic labeling and magnetic guid-ance. Our data conifrm that superparamagnetic iron oxide nanoparticles effectively label bone marrow mesenchymal stem cells and impart sufficient magnetism to respond to the external magnetic ifeld guides. More importantly, superparamagnetic iron oxide-labeled bone marrow mesenchymal stem cells can be dynamically and non-invasively trackedin vivo using magnetic resonance imaging. Superparamagnetic iron oxide labeling of bone marrow mesenchymal stem cells coupled with magnetic guidance offers a promising avenue for the clinical treatment of spinal cord injury.

  4. In vivo monitoring of luciferase labeled bone marrow mesenchymal stem cells by bioluminescent in rats%大鼠活体内 Luciferase 标记骨髓间充质干细胞生物发光示踪监测

    Institute of Scientific and Technical Information of China (English)

    曹娟; 吕琪; 丁辉; 于梦洋; 刘晋阳; 樊毫军

    2016-01-01

    Objective To monitor the colonization and dynamic distribution of allogeneic bone marrow mesenchymal stem cells (BMSC) in healthy rats by using the technique of bioluminescent imaging. Methods BMSC from Wistar rat were infected by lentiviral which carry stable expression genes of luciferase and green fluorescence protein (Luc-GFP-BMSC), then implanted into Wistar rats by tail vein injection. Dynamic distribution of Luc-GFP-BMSC in rats was continuously monitored by using in vivo imaging system. Results In vivo imaging results showed that biological signal was strongest at 1.5 hours after implantation of Luc-GFP-BMSC in rats, and mainly resided in the lung, 24 hours later, signal in the back gradually enhanced and became dominant, after 3 days, signal in the chest couldn’t be detected, after 14 days, biological signal in the whole body disappeared. Conclusions After implantation through the tail vein, allogeneic BMSC in healthy Wistar rats initially reside in the lungs, and then egress to the back, finally the biological signal gradually disappeared, which suggests that tail vein injection of BMSC may be an effective approach for lung disease.%目的:应用生物发光技术监测同种异体骨髓间充质干细胞(bone marrow mesenchymal stem cells, BMSC)在健康大鼠体内的定植与动态分布情况。方法 Wistar 大鼠 BMSC 经稳定表达增强绿色荧光蛋白(enhanced green fluorescent protein, e-GFP)和荧光素酶(luciferase)基因的慢病毒感染后,经尾静脉注射植入 Wistar 大鼠体内,通过活体成像连续监测其动态分布。结果活体成像结果显示,植入感染病毒的骨髓间充质干细胞(Luc-GFP-BMSC)1.5 h 后在大鼠体内生物信号最强,主要聚集在肺,24 h 后背部信号增强并居于主导,第3天胸部信号检测不到,14 d 后,全身生物信号消失。结论同种异体大鼠 BMSC 经尾静脉植入后,首先聚集于肺部,弥散分布于背部,最终信

  5. Mesenchymal dental pulp cells attenuate dentin resorption in homeostasis.

    Science.gov (United States)

    Zheng, Y; Chen, M; He, L; Marão, H F; Sun, D M; Zhou, J; Kim, S G; Song, S; Wang, S L; Mao, J J

    2015-06-01

    Dentin in permanent teeth rarely undergoes resorption in development, homeostasis, or aging, in contrast to bone that undergoes periodic resorption/remodeling. The authors hypothesized that cells in the mesenchymal compartment of dental pulp attenuate osteoclastogenesis. Mononucleated and adherent cells from donor-matched rat dental pulp (dental pulp cells [DPCs]) and alveolar bone (alveolar bone cells [ABCs]) were isolated and separately cocultured with primary rat splenocytes. Primary splenocytes readily aggregated and formed osteoclast-like cells in chemically defined osteoclastogenesis medium with 20 ng/mL of macrophage colony-stimulating factor (M-CSF) and 50 ng/mL of receptor activator of nuclear factor κB ligand (RANKL). Strikingly, DPCs attenuated osteoclastogenesis when cocultured with primary splenocytes, whereas ABCs slightly but significantly promoted osteoclastogenesis. DPCs yielded ~20-fold lower RANKL expression but >2-fold higher osteoprotegerin (OPG) expression than donor-matched ABCs, yielding a RANKL/OPG ratio of 41:1 (ABCs:DPCs). Vitamin D3 significantly promoted RANKL expression in ABCs and OPG in DPCs. In vivo, rat maxillary incisors were atraumatically extracted (without any tooth fractures), followed by retrograde pulpectomy to remove DPCs and immediate replantation into the extraction sockets to allow repopulation of the surgically treated root canal with periodontal and alveolar bone-derived cells. After 8 wk, multiple dentin/root resorption lacunae were present in root dentin with robust RANKL and OPG expression. There were areas of dentin resoprtion alternating with areas of osteodentin formation in root dentin surface in the observed 8 wk. These findings suggest that DPCs of the mesenchymal compartment have an innate ability to attenuate osteoclastogenesis and that this innate ability may be responsible for the absence of dentin resorption in homeostasis. Mesenchymal attenuation of dentin resorption may have implications in internal

  6. The distribution and effect of allogenic mesenchymal stem cells on joints of collagen induced arthritis rats%移植间充质干细胞在胶原诱导性关节炎大鼠关节的分布和作用

    Institute of Scientific and Technical Information of China (English)

    马丽辉; 乔振华; 李小峰

    2011-01-01

    Objective To study the distribution of allogenic bone marrow-derived mesenchymal stem cells (BM-MSCs) on joints of collagen-induced arthritis (CIA) rats and to investigate their repair effects on joint damages. Methods Five Wistar rats were used for extraction of mesenchymal stem cells and 30 adult female Wistar rats were divided into 3 groups: the CIA rats group A (n=10), CIA rats group B (n=10) and normal rats control group C (n=10). BM-MSCs of Wistar rats were isolated, cultured in vitro routinely and the fourth passages was taken for identification of specific surface antigens by flow cytometry, then the cells were labeled with 5-bromodeoxyuridine (5-BrdU) in vitro. The models of CIA rats were established. 5-BrdU labeled BM-MSCs (1.0×107 cells/kg) were imfused from through tail vein to CIA rats group A and control group C. During the first 4 weeks after BM-MSCs transplantation, changes of general condition and left hind paw swelling were examined. At the fourth week, immunohistochemical examination of 5 -BrdU and osteoprotegerin (OPG) were performed to investigate BM-MSCs aggregation around the knee joints. The contribution of BM -MSCs to repairing of joint damages was identified. Comparisons between groups were performed by t-test. Results After BM-MSCs transplantation, left hindpaw swelling of group A were relieved compared with group B (P<0.05) and the mobility of the joints was significantly improved. At the fourth week, much more implanted cells (5-BrdU positive cells.) were detected in the damaged knee joints than those in normal knee joints. The average grey scale values on synovium of knee joints in the CIA group A (85±9) was significantly lower than that of the normal group C (110±6, P<0.05). At the same time, OPG expression was increased in damaged knee joints. The average grey scale values on synovium of knee joints in CIA group A (54±4) was significantly lower than that of the CIA group B (77±6, P<0.05). Conclusion The transplanted

  7. Transplantation of bone marrow mesenchymal stem cells improves the ability of learning and memory of a rat model of Alzheimer's disease%骨髓间充质干细胞移植对老年性痴呆大鼠学习和记忆能力的影响

    Institute of Scientific and Technical Information of China (English)

    牟心红; 张岭; 张玥; 张文成; 李海生

    2012-01-01

    [Objective]To evaluate the effect of bone marrow mesenchymal stem cell (BMSC) transplantation on the ability of learning and memory in Alzheimer's disease (AD) rats. [Methods] The natural senile AD rat model was chosen from the healthy male Wistar rats aged 24 months. The AD rats were randomly divided into three groups with ten animals in each group. In the control group, rats were injected bilaterally with physiological saline into the hippocampus. In the BMSC group, rats received BMSCs. In the differentiation group, rats received injection of neuronal induced BMSCs. The learning ability of the AD rats was detected by Y type maze test one day before and 6 week after operation. The memory ability was tested 48 hours after learning test. The spatial learning and memory ability of rats was evaluated with Morris water maze. The expression localization of choline acetyltransferase (ChAT) in the hippocampus of ratsThe learning and memory scores decreased in control group and increased in BMSC transplantation group, there were all not statistics significance compared with that before transplant treatment (P > 0.05). The learning and memory scores were higher than before in differentiation group (P < 0.01). There was significant difference between control group and the other groups (P < 0.05). The escape latency of hidden platform in differentiation group decreased obviously as compared with control group (P < 0.01). The ChAT-positive neurons were observed in the hippocampus of rats of the differentiation group.[Conclusions]BMSC transplantation may improve the learning and memory ability of AD rats. Neuronal induced BMSC transplantation is superior to undifferentiated BMSCs. Our results demonstrate that BMSCs can play an important role in treating cognitive disturbance of AD rats.%[目的]本研究旨在探索骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)移植对老年性痴呆大鼠学习和记忆的影响.[方法]选用24月龄健康雄性Wistar大

  8. Plant flavonol isorhamnetin attenuates chemically induced inflammatory bowel disease via a PXR-dependent pathway.

    Science.gov (United States)

    Dou, Wei; Zhang, Jingjing; Li, Hao; Kortagere, Sandhya; Sun, Katherine; Ding, Lili; Ren, Gaiyan; Wang, Zhengtao; Mani, Sridhar

    2014-09-01

    Isorhamnetin is an O-methylated flavonol present in fruit and vegetables. We recently reported the identification of isorhamnetin as an activator of the human pregnane X receptor (PXR), a known target for abrogating inflammation in inflammatory bowel disease (IBD). The current study investigated the role of isorhamnetin as a putative mouse PXR activator in ameliorating chemically induced IBD. Using two different models (ulcerative colitis like and Crohn's disease like) of experimental IBD in mice, we demonstrated that isorhamnetin abrogated inflammation through inhibiting the activity of myeloperoxidase, the levels of TNF-α and IL-6, the mRNA expression of proinflammatory mediators (iNOS, ICAM-1, COX2, TNF-α, IL-2 and IL-6) and the phosphorylation of IκBα and NF-κB p65. PXR gene overexpression inhibited NF-κB luciferase activity, and the inhibition was potentiated by isorhamnetin treatment. PXR knockdown by siRNA demonstrated the necessity for PXR in isorhamnetin-mediated up-regulation of xenobiotic metabolism genes. Ligand pocket-filling mutants (S247W/C284W and S247W/C284W/S208W) of human PXR weakened the effect of isorhamnetin on PXR activation. Molecular docking studies and time-resolved fluorescence resonance energy transfer competitive binding assays confirmed the ligand (isorhamnetin)-binding affinity. These results clearly demonstrated the ameliorating effect of isorhamnetin on experimental IBD via PXR-mediated up-regulation of xenobiotic metabolism and down-regulation of NF-κB signaling. The novel findings may contribute to the effective utilization of isorhamnetin or its derivatives as a PXR ligand in the treatment of human IBD.

  9. Bone marrow-derived mesenchymal stem cells increase dopamine synthesis in the injured striatum

    Institute of Scientific and Technical Information of China (English)

    Yue Huang; Cheng Chang; Jiewen Zhang; Xiaoqun Gao

    2012-01-01

    Previous studies showed that tyrosine hydroxylase or neurturin gene-modified cells transplanted into rats with Parkinson's disease significantly improved behavior and increased striatal dopamine content. In the present study, we transplanted tyrosine hydroxylase and neurturin gene-modified bone marrow-derived mesenchymal stem cells into the damaged striatum of Parkinson's disease model rats. Several weeks after cell transplantation, in addition to an improvement of motor function, tyrosine hydroxylase and neurturin proteins were up-regulated in the injured striatum, and importantly, levels of dopamine and its metabolite 3,4-dihydroxyphenylacetic acid increased significantly. Furthermore, the density of the D2 dopamine receptor in the postsynaptic membranes of dopaminergic neurons was decreased. These results indicate that transplantation of tyrosine hydroxylase and neurturin gene-modified bone marrow-derived mesenchymal stem cells increases dopamine synthesis and significantly improves the behavior of rats with Parkinson's disease.

  10. 骨髓间充质干细胞移植对老年性痴呆大鼠空间学习记忆能力的影响%Effects of transplantation of bone marrow mesenchymal stem cells on spacial learning-dysmnesin of Alzheimer's disease in rats

    Institute of Scientific and Technical Information of China (English)

    王金华; 王世民; 孔繁明; 张文治; 唐帆; 张学斌; 苏心; 王新平

    2011-01-01

    Objective: To study the effects of transplantation of bone marrow mesenchymal stem cells (BMSCs) on learning and memory ability and its mechanism in rats with Alzheimer's disease induced by amyloid-β1-40 (Aβ1-40) injection.Method: AD model was induced by injecting Aβ1-40 into bilateral hippocampus. Three weeks later, the bone marrow mesenchymal stem cells was implanted. The learning and memory abilities of rats were evaluated with Morris water maze.Rats were sacrificed after 2 months. Bromodeoxyuridine(BrdU)+neuron specific enolase(NSE), BrdU+glial fibrillary acidic protein (GFAP) immunohistochemistry double-staining cells, and the migration of doublecortin (DCX, the neuron precursors marker) from subventricular zone to the damaged hippocampus were observed.Result:The escape latency of hidden platform in AD group delayed significantly(P<0.05),the escape latency in transplantation treatment group decreased obviously as compared with model group(P<0.05). The immunohistochemistry double-staining cells could be found at the center of damaged site(bilateral hippocampus) and the surrounding sites in rats of transplantation group, the migration of doublecortin from subventricular zone to the damaged hippocampus wasn't observed.Conclusion:The implantation of BMSCs maybe have the effect of improving spacial learning-dysmnesia of AD rats.%目的:研究骨髓间充质干细胞(BMSCs)移植对淀粉样蛋白-β(1-40)(Aβ(1-40))诱导的老年性痴呆(Alzheimer's disease,AD)模型大鼠行为学的影响,并探讨其治疗AD的可能机制.方法:采用向大鼠双侧海马立体定向注射Aβ(1-40)诱导AD动物模型,造模后3周采用同样方法移植骨髓间充质干细胞,采用Morris水迷宫观察大鼠行为学变化.造模后2个月处死大鼠,观察海马组织有无溴脱氧尿嘧啶核苷(Brdu)+神经元特异性烯醇化酶(NSE)、BrdU+胶质纤维酸性蛋白(GFAP)免疫组织化学双染阳性细胞,并且观察从侧脑室到海马是否存在神

  11. Hydroxyapatite incorporated into collagen gels for mesenchymal stem cell culture.

    Science.gov (United States)

    Laydi, F; Rahouadj, R; Cauchois, G; Stoltz, J-F; de Isla, N

    2013-01-01

    Collagen gels could be used as carriers in tissue engineering to improve cell retention and distribution in the defect. In other respect hydroxyapatite could be added to gels to improve mechanical properties and regulate gel contraction. The aim of this work was to analyze the feasibility to incorporate hydroxyapatite into collagen gels and culture mesenchymal stem cells inside it. Human bone marrow mesenchymal stem cells (hMSC-BM) were used in this study. Gels were prepared by mixing rat tail type I collagen, hydroxyapatite microparticles and MSCs. After polymerization gels were kept in culture while gel contraction and mechanical properties were studied. In parallel, cell viability and morphology were analyzed. Gels became free-floating gels contracted from day 3, only in the presence of cells. A linear rapid contraction phase was observed until day 7, then a very slow contraction phase took place. The incorporation of hydroxyapatite improved gel stability and mechanical properties. Cells were randomly distributed on the gel and a few dead cells were observed all over the experiment. This study shows the feasibility and biocompatibility of hydroxyapatite supplemented collagen gels for the culture of mesenchymal stem cells that could be used as scaffolds for cell delivery in osteoarticular regenerative medicine.

  12. Combination of high performance refractometry and infrared spectroscopy as a probe for chemically induced gelation and vitrification of epoxies

    OpenAIRE

    Müller, Ulrich; Philipp, Martine; Gervais, P. C.; Possart, Prof Dr Wulff; Wehlack, C.; Kieffer, J.; Sanctuary, Roland; Krüger, Jan-Kristian

    2010-01-01

    A combination of infrared spectroscopy and high performance refractometry was used to investigate the chemically induced sol-gel and glass transition during the polymerization of epoxies. Representations of the refractive index versus chemical conversion reveal an interesting insight in the optical properties accompanying gelation and vitrification. Whereas the electronic polarizability of the liquid state of small average molecular mass and the glassy state is dominated by the mass density, ...

  13. Evaluation of anticonvulsant activity of volatile oil extract of Nigella sativa seeds by chemically induced seizure model in albino rats

    Directory of Open Access Journals (Sweden)

    Asmatanzeem Bepari

    2016-08-01

    Conclusions: The N. sativa seeds showed anticonvulsant activity in pentylenetetrazole induced seizure model of epilepsy. This study showed that volatile oil of N. sativa seeds potentiated the effect of sodium valproate. [Int J Basic Clin Pharmacol 2016; 5(4.000: 1300-1307

  14. Effects of Metformin, Buformin, and Phenformin on the Post Initiation Stage of Chemically-Induced Mammary Carcinogenesis in the Rat

    OpenAIRE

    2015-01-01

    Metformin is a widely prescribed drug for the treatment of type-2 diabetes. Although epidemiological data have provided a strong rationale for investigating the potential of this biguanide for use in cancer prevention and control, uncertainty exists whether metformin should be expected to have an impact in non-diabetic patients. Furthermore, little attention has been given to the possibility that other biguanides may have anticancer activity. In this study, the effects of clinically relevant ...

  15. Role of the Slug Transcription Factor in Chemically-Induced Skin Cancer

    Directory of Open Access Journals (Sweden)

    Kristine von Maltzan

    2016-02-01

    Full Text Available The Slug transcription factor plays an important role in ultraviolet radiation (UVR-induced skin carcinogenesis, particularly in the epithelial-mesenchymal transition (EMT occurring during tumor progression. In the present studies, we investigated the role of Slug in two-stage chemical skin carcinogenesis. Slug and the related transcription factor Snail were expressed at high levels in skin tumors induced by 7,12-dimethylbenz[α]anthracene application followed by 12-O-tetradecanoylphorbol-13-acetate (TPA treatment. TPA-induced transient elevation of Slug and Snail proteins in normal mouse epidermis and studies in Slug transgenic mice indicated that Slug modulates TPA-induced epidermal hyperplasia and cutaneous inflammation. Although Snail family factors have been linked to inflammation via interactions with the cyclooxygenase-2 (COX-2 pathway, a pathway that also plays an important role in skin carcinogenesis, transient TPA induction of Slug and Snail appeared unrelated to COX-2 expression. In cultured human keratinocytes, TPA induced Snail mRNA expression while suppressing Slug expression, and this differential regulation was due specifically to activation of the TPA receptor. These studies show that Slug and Snail exhibit similar patterns of expression during both UVR and chemical skin carcinogenesis, that Slug and Snail can be differentially regulated under some conditions and that in vitro findings may not recapitulate in vivo results.

  16. Adenovirus-mediated human brain-derived neurotrophic factor gene-modified bone marrow mesenchymal stem cell transplantation for spinal cord injury

    Institute of Scientific and Technical Information of China (English)

    Changsheng Wang; Jianhua Lin; Chaoyang Wu; Rongsheng Chen

    2011-01-01

    Rat bone marrow mesenchymal stem cells expressing brain-derived neurotrophic factor were successfully obtained using a gene transfection method, then intravenously transplanted into rats with spinal cord injury. At 1, 3, and 5 weeks after transplantation, the expression of ??brain-derived neurotrophic factor and neurofilament-200 was upregulated in the injured spinal cord, spinal cord injury was alleviated, and Basso-Beattie-Bresnahan scores of hindlimb motor function were significantly increased. This evidence suggested that intravenous transplantation of adenovirus- mediated brain-derived neurotrophic factor gene-modified rat bone marrow mesenchymal stem cells could play a dual role, simultaneously providing neural stem cells and neurotrophic factors.

  17. B-lymphocytes as key players in chemical-induced asthma.

    Directory of Open Access Journals (Sweden)

    Vanessa De Vooght

    Full Text Available T-lymphocytes and B-lymphocytes are key players in allergic asthma, with B-lymphocytes producing antigen-specific immunoglobulins E (IgE. We used a mouse model of chemical-induced asthma and transferred B-lymphocytes from sensitized animals into naïve wild type mice, B-lymphocyte knock-out (B-KO mice or severe combined immunodeficiency (SCID mice. On days 1 and 8, BALB/c mice were dermally sensitized with 0.3% toluene diisocyanate (TDI (20 µl/ear. On day 15, mice were euthanized and the auricular lymph nodes isolated. B-lymphocytes (CD19(+ were separated from the whole cell suspension and 175,000 cells were injected in the tail vein of naïve wild type, B-KO or SCID mice. Three days later, the mice received a single oropharyngeal challenge with 0.01% TDI (20 µl or vehicle (acetone/olive oil (AOO (controls. Airway reactivity to methacholine and total and differential cell counts in the bronchoalveolar lavage (BAL fluid were measured 24 hours after challenge. B-lymphocytes of AOO or TDI-sensitized mice were characterized for the expression of surface markers and production of cytokines. We found that transfer of B-cells obtained from mice dermally sensitized to toluene diisocyanate (TDI into naïve wild type mice, B-KO mice or SCID mice led, within three days, to an acute asthma-like phenotype after an airway challenge with TDI. This response was specific and independent of IgE. These B-lymphocytes showed antigen presenting capacities (CD80/CD86 and CD40 and consisted of B effector (Be2- (IL-4 and Be1-lymphocytes (IFN-γ. The transferred B-lymphocytes were visualized near large airways, 24 hours after TDI challenge. Thus, B-lymphocytes can provoke an asthmatic response without the action of T-lymphocytes and without major involvement of IgE.

  18. 骨髓间充质干细胞黑质内移植对帕金森病大鼠的治疗作用%Therapeutic effects of transplantation of bone marrow mesenchymal stem cells into substantia nigra on the rat model of Parkinson's disease

    Institute of Scientific and Technical Information of China (English)

    陈丹丹; 付文玉; 庄宝祥; 李锋杰; 吕翠

    2013-01-01

    目的 探索骨髓间充质干细胞(BMSCs)移植到帕金森病(Parkinson's disease,PD)大鼠毁损侧黑质内,PD模型大鼠的姿势不对称性和黑质及纹状体内酪氨酸羟化酶(tyrosine hydroxylase,TH)表达的改变,以及BMSCs在大鼠脑内的存活、分化情况.方法 黑质、前脑内侧束两点法注射6-羟多巴胺(6-OHDH)并行为学分析筛选PD模型大鼠.将PD模型大鼠随机分为移植组和对照组.BMSCs移植术后4周和8周,观察大鼠姿势不对称性,免疫组织化学及免疫荧光显色方法检测黑质和纹状体酪氨酸羟化酶 (tyrosine hydroxylase,TH) 的表达变化以及BMSCs在大鼠体内的存活、迁移及分化情况.结果 BMSCs黑质内移植可使PD模型大鼠的转动频率由(10.62±2.97)r/min降至(4.65±1.08)r/min(P<0.01),显著增加毁损侧黑质TH阳性细胞数量和纹状体内TH阳性纤维密度.BMSCs在大鼠黑质内可以存活至少8周,部分细胞分化为神经干细胞、神经元和神经胶质细胞.结论 黑质内移植BMSCs对PD模型大鼠有一定的治疗作用.%Objective To explore the effect of bone marrow mesenchymal stem cells (BMSCs) transplanted into the unilateral lesioned substantia nigra (SN) , the changes of postural asymmetry and expression of tyrosine hydroxylase in SN of Parkinson's disease ( PD) model rats, and the survival and differentiation of BMSCs in the brain tissue. Methods Model of Parkinson's dieases (PD) was induced by 6-hydroxydopamine (6-OHDA) injection into the right SN and medial forebrain bundle in the rat brains. The PD rats were randomly divided into transplantation group and control group. The PD rat rotational behavior was induced by apomorphine ( APO) at 4 weeks and 8 weeks after BMSCs transplantation. The rat behavior was tested, and the expression of tyrosine hydroxylase ( TH ) and the survival, migration and differentiation of transplanted BMSCs in the SN and ST were examined using immunohistochemical and immunofluorescence

  19. Overexpression of microRNA-124 promotes the neuronal differentiation of bone marrow-derived mesenchymal stem cells

    Institute of Scientific and Technical Information of China (English)

    Defeng Zou; Yi Chen; Yaxin Han; Chen Lv; Guanjun Tu

    2014-01-01

    microRNAs (miRNAs) play an important regulatory role in the self-renewal and differentiation of stem cells. In this study, we examined the effects of miRNA-124 (miR-124) overexpression in bone marrow-derived mesenchymal stem cells. In particular, we focused on the effect of overexpression on the differentiation of bone marrow-derived mesenchymal stem cells into neurons. First, we used GeneChip technology to analyze the expression of miRNAs inbone marrow-derived mesen-chymal stem cells, neural stem cells and neurons. miR-124 expression was substantially reduced inbone marrow-derived mesenchymal stem cells compared with the other cell types. We con-structed a lentiviral vector overexpressing miR-124 and transfected it intobone marrow-derived mesenchymal stem cells. Intracellular expression levels of the neuronal early markersβ-III tu-bulin and microtubule-associated protein-2 were signiifcantly increased, and apoptosis induced by oxygen and glucose deprivation was reduced in transfected cells. After miR-124-transfected bone marrow-derived mesenchymal stem cells were transplanted into the injured rat spinal cord, a large number of cells positive for the neuronal marker neurofilament-200 were observed in the transplanted region. The Basso-Beattie-Bresnahan locomotion scores showed that the motor function of the hind limb of rats with spinal cord injury was substantially improved. These re-sults suggest that miR-124 plays an important role in the differentiation ofbone marrow-derived mesenchymal stem cells into neurons. Our ifndings should facilitate the development of novel strategies for enhancing the therapeutic efifcacy ofbone marrow-derived mesenchymal stem cell transplantation for spinal cord injury.

  20. 660 nm red light-enhanced bone marrow mesenchymal stem cell transplantation for hypoxic-ischemic brain damage treatment

    Institute of Scientific and Technical Information of China (English)

    Xianchao Li; Wensheng Hou; Xiaoying Wu; Wei Jiang; Haiyan Chen; Nong Xiao; Ping Zhou

    2014-01-01

    Bone marrow mesenchymal stem cell transplantation is an effective treatment for neonatal hy-poxic-ischemic brain damage. However, the in vivo transplantation effects are poor and their survival, colonization and differentiation efifciencies are relatively low. Red or near-infrared light from 600-1,000 nm promotes cellular migration and prevents apoptosis. Thus, we hypothesized that the combination of red light with bone marrow mesenchymal stem cell transplantation would be effective for the treatment of hypoxic-ischemic brain damage. In this study, the migra-tion and colonization of cultured bone marrow mesenchymal stem cells on primary neurons after oxygen-glucose deprivation were detected using Transwell assay. The results showed that, after a 40-hour irradiation under red light-emitting diodes at 660 nm and 60 mW/cm2, an increasing number of green lfuorescence-labeled bone marrow mesenchymal stem cells migrated towards hypoxic-ischemic damaged primary neurons. Meanwhile, neonatal rats with hypoxic-ischemic brain damage were given an intraperitoneal injection of 1 × 106 bone marrow mesenchymal stem cells, followed by irradiation under red light-emitting diodes at 660 nm and 60 mW/cm2 for 7 successive days. Shuttle box test results showed that, after phototherapy and bone marrow mesenchymal stem cell transplantation, the active avoidance response rate of hypoxic-ischemic brain damage rats was significantly increased, which was higher than that after bone marrow mesenchymal stem cell transplantation alone. Experimental ifndings indicate that 660 nm red light emitting diode irradiation promotes the migration of bone marrow mesenchymal stem cells, thereby enhancing the contribution of cell transplantation in the treatment of hypox-ic-ischemic brain damage.

  1. 660 nm red light-enhanced bone marrow mesenchymal stem cell transplantation for hypoxic-ischemic brain damage treatment.

    Science.gov (United States)

    Li, Xianchao; Hou, Wensheng; Wu, Xiaoying; Jiang, Wei; Chen, Haiyan; Xiao, Nong; Zhou, Ping

    2014-02-01

    Bone marrow mesenchymal stem cell transplantation is an effective treatment for neonatal hypoxic-ischemic brain damage. However, the in vivo transplantation effects are poor and their survival, colonization and differentiation efficiencies are relatively low. Red or near-infrared light from 600-1,000 nm promotes cellular migration and prevents apoptosis. Thus, we hypothesized that the combination of red light with bone marrow mesenchymal stem cell transplantation would be effective for the treatment of hypoxic-ischemic brain damage. In this study, the migration and colonization of cultured bone marrow mesenchymal stem cells on primary neurons after oxygen-glucose deprivation were detected using Transwell assay. The results showed that, after a 40-hour irradiation under red light-emitting diodes at 660 nm and 60 mW/cm(2), an increasing number of green fluorescence-labeled bone marrow mesenc