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Sample records for human acellular matrix

  1. Data from acellular human heart matrix

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    Pedro L Sánchez

    2016-09-01

    Full Text Available Perfusion decellularization of cadaveric hearts removes cells and generates a cell-free extracellular matrix scaffold containing acellular vascular conduits, which are theoretically sufficient to perfuse and support tissue-engineered heart constructs. This article contains additional data of our experience decellularizing and testing structural integrity and composition of a large series of human hearts, “Acellular human heart matrix: a critical step toward whole heat grafts” (Sanchez et al., 2015 [1]. Here we provide the information about the heart decellularization technique, the valve competence evaluation of the decellularized scaffolds, the integrity evaluation of epicardial and myocardial coronary circulation, the pressure volume measurements, the primers used to assess cardiac muscle gene expression and, the characteristics of donors, donor hearts, scaffolds and perfusion decellularization process.

  2. Human acellular dermal wound matrix: evidence and experience.

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    Kirsner, Robert S; Bohn, Greg; Driver, Vickie R; Mills, Joseph L; Nanney, Lillian B; Williams, Marie L; Wu, Stephanie C

    2015-12-01

    A chronic wound fails to complete an orderly and timely reparative process and places patients at increased risk for wound complications that negatively impact quality of life and require greater health care expenditure. The role of extracellular matrix (ECM) is critical in normal and chronic wound repair. Not only is ECM the largest component of the dermal skin layer, but also ECM proteins provide structure and cell signalling that are necessary for successful tissue repair. Chronic wounds are characterised by their inflammatory and proteolytic environment, which degrades the ECM. Human acellular dermal matrices, which provide an ECM scaffold, therefore, are being used to treat chronic wounds. The ideal human acellular dermal wound matrix (HADWM) would support regenerative healing, providing a structure that could be repopulated by the body's cells. Experienced wound care investigators and clinicians discussed the function of ECM, the evidence related to a specific HADWM (Graftjacket(®) regenerative tissue matrix, Wright Medical Technology, Inc., licensed by KCI USA, Inc., San Antonio, TX), and their clinical experience with this scaffold. This article distills these discussions into an evidence-based and practical overview for treating chronic lower extremity wounds with this HADWM. © 2013 The Authors. International Wound Journal © 2013 Medicalhelplines.com Inc and John Wiley & Sons Ltd.

  3. Coverage of Megaprosthesis with Human Acellular Dermal Matrix after Ewing's Sarcoma Resection: A Case Report

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    Robert M. Whitfield

    2011-01-01

    Full Text Available A 23-year-old female with Ewing's Sarcoma underwent tibial resection and skeletal reconstruction using proximal tibial allograft prosthetic reconstruction with distal femur endoprosthetic reconstruction and rotating hinge. Human acellular dermal matrix, (Alloderm, LifeCell, Branchburg, NJ, USA, was used to wrap the skeletal reconstruction. Soft tissue reconstruction was completed with a rotational gastrocnemius muscle flap and skin graft. Despite prolonged immobilization, the patient quickly regained full range of motion of her skeletal reconstruction. Synthetic mesh, tapes and tubes are used to perform capsule reconstruction of megaprosthesis. This paper describes the role of human acellular dermal matrix in capsule reconstruction around a megaprosthesis.

  4. Coverage of Megaprosthesis with Human Acellular Dermal Matrix after Ewing's Sarcoma Resection: A Case Report.

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    Whitfield, Robert M; Rinard, Jeremy; King, David

    2011-01-01

    A 23-year-old female with Ewing's Sarcoma underwent tibial resection and skeletal reconstruction using proximal tibial allograft prosthetic reconstruction with distal femur endoprosthetic reconstruction and rotating hinge. Human acellular dermal matrix, (Alloderm, LifeCell, Branchburg, NJ, USA), was used to wrap the skeletal reconstruction. Soft tissue reconstruction was completed with a rotational gastrocnemius muscle flap and skin graft. Despite prolonged immobilization, the patient quickly regained full range of motion of her skeletal reconstruction. Synthetic mesh, tapes and tubes are used to perform capsule reconstruction of megaprosthesis. This paper describes the role of human acellular dermal matrix in capsule reconstruction around a megaprosthesis.

  5. A new candidate substrate for cell-matrix adhesion study: the acellular human amniotic matrix.

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    Guo, Qianchen; Lu, Xuya; Xue, Yuan; Zheng, Hong; Zhao, Xiaotao; Zhao, Huajian

    2012-01-01

    In vivo adhesions between cells and the extracellular matrix play a crucial role in cell differentiation, proliferation, and migration as well as tissue remodeling. Natural three-dimensional (3D) matrices, such as self-assembling matrices and Matrigel, have limitations in terms of their biomechanical properties. Here, we present a simple method to produce an acellular human amniotic matrix (AHAM) with preserved biomechanical properties and a favorable adhesion potential. On the stromal side of the AHAM, human foreskin fibroblasts (HFFs) attached and extended with bipolar spindle-shaped morphology proliferated to multilayer networks, invaded into the AHAM, and migrated in a straight line. Moreover, αV integrin, paxillin, and fibronectin were observed to colocalize after 24 h of HFF culture on the stromal side of the AHAM. Our results indicate that the AHAM may be an ideal candidate as a cell-matrix adhesion substrate to study cell adhesion and invasion as well as other functions in vitro under a tensile force that mimics the in vivo environment.

  6. A New Candidate Substrate for Cell-Matrix Adhesion Study: The Acellular Human Amniotic Matrix

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    Qianchen Guo

    2012-01-01

    Full Text Available In vivo adhesions between cells and the extracellular matrix play a crucial role in cell differentiation, proliferation, and migration as well as tissue remodeling. Natural three-dimensional (3D matrices, such as self-assembling matrices and Matrigel, have limitations in terms of their biomechanical properties. Here, we present a simple method to produce an acellular human amniotic matrix (AHAM with preserved biomechanical properties and a favorable adhesion potential. On the stromal side of the AHAM, human foreskin fibroblasts (HFFs attached and extended with bipolar spindle-shaped morphology proliferated to multilayer networks, invaded into the AHAM, and migrated in a straight line. Moreover, αV integrin, paxillin, and fibronectin were observed to colocalize after 24 h of HFF culture on the stromal side of the AHAM. Our results indicate that the AHAM may be an ideal candidate as a cell-matrix adhesion substrate to study cell adhesion and invasion as well as other functions in vitro under a tensile force that mimics the in vivo environment.

  7. Production of an acellular matrix from amniotic membrane for the synthesis of a human skin equivalent.

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    Sanluis-Verdes, Anahí; Yebra-Pimentel Vilar, Maria Teresa; García-Barreiro, Juan Javier; García-Camba, Marta; Ibáñez, Jacinto Sánchez; Doménech, Nieves; Rendal-Vázquez, Maria Esther

    2015-09-01

    Human amniotic membrane (HAM) has useful properties as a dermal matrix substitute. The objective of our work was to obtain, using different enzymatic or chemical treatments to eliminate cells, a scaffold of acellular HAM for later use as a support for the development of a skin equivalent. The HAM was separated from the chorion, incubated and cryopreserved. The membrane underwent different enzymatic and chemical treatments to eliminate the cells. Fibroblasts and keratinocytes were separately obtained from skin biopsies of patients following a sequential double digestion with first collagenase and then trypsin-EDTA (T/E). A skin equivalent was then constructed by seeding keratinocytes on the epithelial side and fibroblasts on the chorionic side of the decellularizated HAM. Histological, immunohistochemical, inmunofluorescent and molecular biology studies were performed. Treatment with 1% T/E at 37 °C for 30 min totally removed epithelial and mesenchymal cells. The HAM thus treated proved to be a good matrix to support adherence of cells and allowed the achievement of an integral and intact scaffold for development of a skin equivalent, which could be useful as a skin substitute for clinical use.

  8. Human acellular dermal matrix for repair of abdominal wall defects: review of clinical experience and experimental data.

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    Holton, Luther H; Kim, Daniel; Silverman, Ronald P; Rodriguez, Eduardo D; Singh, Navin; Goldberg, Nelson H

    2005-01-01

    The use of prosthetic mesh for the tension-free repair of incisional hernias has been shown to be more effective than primary suture repair. Unfortunately, prosthetic materials can be a suboptimal choice in a variety of clinical scenarios. In general, prosthetic materials should not be implanted into sites with known contamination or infection because they lack an endogenous vascular network and are thus incapable of clearing bacteria. This is of particular relevance to the repair of recurrent hernias, which are often refractory to repair because of indolent bacterial colonization that weakens the site and retards appropriate healing. Although fascia lata grafts and muscle flaps can be employed for tension-free hernia repairs, they carry the potential for significant donor site morbidity. Recently, a growing number of clinicians have used human acellular dermal matrix as a graft material for the tension-free repair of ventral hernias. This material has been shown to become revascularized in both animal and human subjects. Once repopulated with a vascular network, this graft material is theoretically capable of clearing bacteria, a property not found in prosthetic graft materials. Unlike autologous materials such as fascial grafts and muscle flaps, acellular dermal matrix can be used without subjecting the patient to additional morbidity in the form of donor site complications. This article presents a thorough review of the current literature, describing the properties of human acellular dermal matrix and discussing both animal and human studies of its clinical performance. In addition to the review of previously published clinical experiences, we discuss our own preliminary results with the use of acellular dermal matrix for ventral hernia repair in 46 patients.

  9. Chondrogenesis of human infrapatellar fat pad stem cells on acellular dermal matrix

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    Ken eYe

    2016-01-01

    Full Text Available Acellular dermal matrix (ADM has been in clinical use for decades in numerous surgical applications. The ability for ADM to promote cellular repopulation and revascularisation, and tissue regeneration is well documented. Adipose stem cells have the ability to differentiate into mesenchymal tissue types, including bone and cartilage. The aim of this study was to investigate the potential interaction between ADM and adipose stem cells in vitro using TGFβ3 and BMP6.Human infrapatellar fat pad derived adipose stem cells (IPFP-ASC were cultured with ADM derived from rat dermis under chondrogenic (TGFβ3 and BMP6 in vitro for 2 and 4 weeks. Histology, qPCR and immunohistochemistry were performed to assess for markers of chondrogenesis (collagen Type II, SOX9 and proteoglycans. At 4 weeks, cell-scaffold constructs displayed cellular changes consistent with chondrogenesis, with evidence of stratification of cell layers and development of a hyaline-like cartilage layer superficially which stained positively for collagen Type II and proteoglycans. Significant cell-matrix interaction was seen between the cartilage layer and the ADM itself with seamless integration between each layer. Real time qPCR showed significantly increases of COL2A1, SOX9, and ACAN gene expression over 4 weeks when compared to control. COL1A2 gene expression remained unchanged over 4 weeks.We believe the principles which make ADM versatile and successful for tissue regeneration are application to cartilage regeneration. This study demonstrates in vitro the ability for IPFP-ASCs to undergo chondrogenesis, infiltrate and interact with ADM. These outcomes serve as a platform for in vivo modelling of ADM for cartilage repair.

  10. Early escharectomy and concurrent composite skin grafting over human acellular dermal matrix scaffold for covering deep facial burns.

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    Tang, Bing; Zhu, Bin; Liang, Yue-Ying; Bi, Liang-Kuan; Chen, Bin; Hu, Zhi-Cheng; Zhang, Kai; Zhu, Jia-Yuan

    2011-04-01

    Although escharectomy and full-thickness skin autografting have been widely used to treat deep facial burns, the clinical outcomes remain unacceptable. Composite razor-thin skin grafting over acellular dermal matrix scaffold has been used successfully in repairing burns of the trunk and limbs, but its use in covering deep facial burns has rarely been reported. In this study, the authors investigated the clinical outcomes of early escharectomy and concurrent composite razor-thin skin autografting and acellular dermal matrix scaffold for treating deep facial burns. Patients with deep facial burns (n = 16) involving 8 to 30 percent of the total body surface area received early escharectomy by postburn day 3 and concurrent, one-stage, large, razor-thin skin autografting on top of human acellular dermal matrix scaffold. Wound dressings were changed on postoperative days 7, 9, and 12 to examine the survival of skin autografts. Patients were followed up for 12 months to evaluate their facial profiles. The take rate of composite skin autografts was 97.3 percent at postoperative day 12. At the follow-up visit, the skin autografts appeared normal in color, with soft texture and good elasticity. The skin junctures showed little scarring. The patients exhibited a chubby facial appearance and abundant expression, except for one patient with microstomia and two patients with ectropion who required further plastic surgical interventions. Early escharectomy and concurrent composite razor-thin skin autografting on top of acellular dermal matrix scaffold constitute an effective and favorable option for covering deep facial burns, especially for patients with limited donor sites.

  11. The effects of acellular amniotic membrane matrix on osteogenic differentiation and ERK1/2 signaling in human dental apical papilla cells.

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    Chen, Yi-Jane; Chung, Min-Chun; Jane Yao, Chung-Chen; Huang, Chien-Hsun; Chang, Hao-Hueng; Jeng, Jiiang-Huei; Young, Tai-Horng

    2012-01-01

    The amniotic membrane (AM) has been widely used in the field of tissue engineering because of the favorable biological properties for scaffolding material. However, little is known about the effects of an acellular AM matrix on the osteogenic differentiation of mesenchymal stem cells. In this study, it was found that both basement membrane side and collagenous stroma side of the acellular AM matrix were capable of providing a preferential environment for driving the osteogenic differentiation of human dental apical papilla cells (APCs) with proven stem cell characteristics. Acellular AM matrix potentiated the induction effect of osteogenic supplements (OS) such as ascorbic acid, β-glycerophosphate, and dexamethasone and enhanced the osteogenic differentiation of APCs, as seen by increased core-binding factor alpha 1 (Cbfa-1) phosphorylation, alkaline phosphatase activity, mRNA expression of osteogenic marker genes, and mineralized matrix deposition. Even in the absence of soluble OS, acellular AM matrix also could exert the substrate-induced effect on initiating APCs' differentiation. Especially, the collagenous stroma side was more effective than the basement membrane side. Moreover, the AM-induced effect was significantly inhibited by U0126, an inhibitor of extracellular signaling-regulated kinase 1/2 (ERK1/2) signaling. Taken together, the osteogenic differentiation promoting effect on APCs is AM-specific, which provides potential applications of acellular AM matrix in bone/tooth tissue engineering.

  12. Human Keratinocyte Growth and Differentiation on Acellular Porcine Dermal Matrix in relation to Wound Healing Potential

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    Robert Zajicek

    2012-01-01

    Full Text Available A number of implantable biomaterials derived from animal tissues are now used in modern surgery. Xe-Derma is a dry, sterile, acellular porcine dermis. It has a remarkable healing effect on burns and other wounds. Our hypothesis was that the natural biological structure of Xe-Derma plays an important role in keratinocyte proliferation and formation of epidermal architecture in vitro as well as in vivo. The bioactivity of Xe-Derma was studied by a cell culture assay. We analyzed growth and differentiation of human keratinocytes cultured in vitro on Xe-Derma, and we compared the results with formation of neoepidermis in the deep dermal wounds treated with Xe-Derma. Keratinocytes cultured on Xe-Derma submerged in the culture medium achieved confluence in 7–10 days. After lifting the cultures to the air-liquid interface, the keratinocytes were stratified and differentiated within one week, forming an epidermis with basal, spinous, granular, and stratum corneum layers. Immunohistochemical detection of high-molecular weight cytokeratins (HMW CKs, CD29, p63, and involucrin confirmed the similarity of organization and differentiation of the cultured epidermal cells to the normal epidermis. The results suggest that the firm natural structure of Xe-Derma stimulates proliferation and differentiation of human primary keratinocytes and by this way improves wound healing.

  13. Acellular Dermal Matrix in Postmastectomy Breast Reconstruction

    NARCIS (Netherlands)

    A.M.S. Ibrahim (Ahmed)

    2014-01-01

    markdownabstract__Abstract__ Over the last decade the use of acellular dermal matrix (ADM) in reconstructive breast surgery has been transformative. Some authors have gone as far as to suggest that it is the single most important advancement in prosthetic breast reconstruction. ADMs are able

  14. Surgical Outcomes of Deep Superior Sulcus Augmentation Using Acellular Human Dermal Matrix in Anophthalmic or Phthisis Socket.

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    Cho, Won-Kyung; Jung, Su-Kyung; Paik, Ji-Sun; Yang, Suk-Woo

    2016-07-01

    Patients with anophthalmic or phthisis socket suffer from cosmetic problems. To resolve those problems, the authors present the surgical outcomes of deep superior sulcus (DSS) augmentation using acellular dermal matrix in patients with anophthalmic or phthisis socket. The authors retrospectively reviewed anophthalmic or phthisis patients who underwent surgery for DSS augmentation using acellular dermal matrix. To evaluate surgical outcomes, the authors focused on 3 aspects: the possibility of wearing contact prosthesis, the degree of correction of the DSS, and any surgical complications. The degree of correction of DSS was classified as excellent: restoration of superior sulcus enough to remove sunken sulcus shadow; fair: gain of correction effect but sunken shadow remained; or fail: no effect of correction at all. Ten eyes of 10 patients were included. There was a mean 21.3 ± 37.1-month period from evisceration or enucleation to the operation for DSS augmentation. All patients could wear contact prosthesis after the operation (100%). The degree of correction was excellent in 8 patients (80%) and fair in 2. Three of 10 (30%) showed complications: eyelid entropion, upper eyelid multiple creases, and spontaneous wound dehiscence followed by inflammation after stitch removal. Uneven skin surface and paresthesia in the forehead area of the affected eye may be observed after surgery. The overall surgical outcomes were favorable, showing an excellent degree of correction of DSS and low surgical complication rates. This procedure is effective for patients who have DSS in the absence or atrophy of the eyeball.

  15. Cartilage oligomeric matrix protein enhances the vascularization of acellular nerves

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    Wei-ling Cui

    2016-01-01

    Full Text Available Vascularization of acellular nerves has been shown to contribute to nerve bridging. In this study, we used a 10-mm sciatic nerve defect model in rats to determine whether cartilage oligomeric matrix protein enhances the vascularization of injured acellular nerves. The rat nerve defects were treated with acellular nerve grafting (control group alone or acellular nerve grafting combined with intraperitoneal injection of cartilage oligomeric matrix protein (experimental group. As shown through two-dimensional imaging, the vessels began to invade into the acellular nerve graft from both anastomotic ends at day 7 post-operation, and gradually covered the entire graft at day 21. The vascular density, vascular area, and the velocity of revascularization in the experimental group were all higher than those in the control group. These results indicate that cartilage oligomeric matrix protein enhances the vascularization of acellular nerves.

  16. Cartilage oligomeric matrix protein enhances the vascularization of acellular nerves

    Institute of Scientific and Technical Information of China (English)

    Wei-ling Cui; Long-hai Qiu; Jia-yan Lian; Jia-chun Li; Jun Hu; Xiao-lin Liu

    2016-01-01

    Vascularization of acellular nerves has been shown to contribute to nerve bridging. In this study, we used a 10-mm sciatic nerve defect model in rats to determine whether cartilage oligomeric matrix protein enhances the vascularization of injured acellular nerves. The rat nerve defects were treated with acellular nerve grafting (control group) alone or acellular nerve grafting combined with intraperitoneal injection of cartilage oligomeric matrix protein (experimental group). As shown through two-dimensional imaging, the vessels began to invade into the acellular nerve graft from both anastomotic ends at day 7 post-operation, and gradually covered the entire graft at day 21. The vascular density, vascular area, and the velocity of revascularization in the experimental group were all higher than those in the control group. These results indicate that cartilage oligomeric matrix protein enhances the vascularization of acellular nerves.

  17. Ridge preservation with acellular dermal matrix and anorganic bone matrix cell-binding peptide P-15 after tooth extraction in humans. A histologic and morphometric study

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    Arthur B. Novaes Jr

    2012-06-01

    Full Text Available Aim: The aim of this study was to analyze by histomorphometric parameters the use of acellular dermal matrix (ADM with or without anorganic bovine bone matrix (ABM / synthetic cell-binding peptide P-15 in the formation of bone in human alveoli. Materials and methods: Eighteen patients in need of extraction of maxillary anterior teeth were selected and randomly assigned to the test group (ADM plus ABM/P-15 or the control group (ADM only. Histomorphometric measurements and histological analysis were recorded about 6 months after ridge preservation procedures in ten patients. The amount of newly formed bone, the most recently formed bone, fibrous tissue plus marrow spaces and remaining graft particles were measured and analyzed. Results: At 6 months, the new bone area parameter and the percentage of fibrous tissue plus marrow space areas showed higher values to the control group, and statistically significant differences when compared with the test group (p=0.03. Conclusion: The ADM acted as a membrane. The association of ABM/P-15 with ADM resulted in new bone formation within the alveoli, but the results were not considered relevant when used in this indication.

  18. Evaluation of lymphangiogenesis in acellular dermal matrix

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    Mario Cherubino

    2014-01-01

    Full Text Available Introduction: Much attention has been directed towards understanding the phenomena of angiogenesis and lymphangiogenesis in wound healing. Thanks to the manifold dermal substitute available nowadays, wound treatment has improved greatly. Many studies have been published about angiogenesis and cell invasion in INTEGRA® . On the other hand, the development of the lymphatic network in acellular dermal matrix (ADM is a more obscure matter. In this article, we aim to characterize the different phases of host cell invasion in ADM. Special attention was given to lymphangiogenic aspects. Materials and Methods: Among 57 rats selected to analyse the role of ADM in lymphangiogenesis, we created four groups. We performed an excision procedure on both thighs of these rats: On the left one we did not perform any action except repairing the borders of the wound; while on the right one we used INTEGRA® implant. The excision biopsy was performed at four different times: First group after 7 days, second after 14 days, third after 21 days and fourth after 28 days. For our microscopic evaluation, we used the classical staining technique of haematoxylin and eosin and a semi-quantitative method in order to evaluate cellularity counts. To assess angiogenesis and lymphangiogenesis development we employed PROX-1 Ab and CD31/PECAM for immunohistochemical analysis. Results: We found remarkable wound contraction in defects that healed by secondary intention while minor wound contraction was observed in defects treated with ADM. At day 7, optical microscopy revealed a more plentiful cellularity in the granulation tissue compared with the dermal regeneration matrix. The immunohistochemical process highlighted vascular and lymphatic cells in both groups. After 14 days a high grade of fibrosis was noticeable in the non-treated group. At day 21, both lymphatic and vascular endothelial cells were better developed in the group with a dermal matrix application. At day 28

  19. Inguinal hernia repair using human acellular dermal matrix%脱细胞真皮基质修补腹股沟疝

    Institute of Scientific and Technical Information of China (English)

    刘飞德; 李基业; 姚胜; 王世斌; 朱瑛梅

    2011-01-01

    BACKGROUND: Tension-free repair using polypropylene mesh is the standard technique for inguinal hernia repair at the present,but the prosthetic material maybe has harmful impact on the patient reproductive functions.OBJECTIVE: To summarize the experience and evaluate the clinical effect of human acellular dermal matrix in inguinal hernia repair.METHODS: Nineteen patients aged 5-38 years with inguinal hernia underwent hernia repair using human acelluar demall matrix.Of the patients, there were 15 male and 4 female. The wound healing was observed and regular follow-up was conducted.RESULTS AND CONCLUSION: Of the 19 patients, all patients recovered with primary wound healing without infection of incisional wound or seroma. Eighteen patients were followed up. During a follow-up of 3-30 months, no chronic pain or discomfort at the incisional area or hernia recurrence occurred. It is feasible and safe to use human acellular dermal matrix patch in inguinal hernia repair, especially in young people or man with inguinal hernia willing to procreate.%背景:当前应用聚丙烯补片行腹股沟疝无张力修补已成为腹股沟疝修补的标准手段,但这些材料可能对患者生殖功能产生影响.目的:总结应用脱细胞真皮基质修补腹股沟疝的经验.方法:回顾性分析19例应用异体脱细胞真皮基质修补腹股沟疝患者的临床资料,男15例,女4例,年龄5~38岁.术后观察切口愈合情况,并定期随访.结果与结论:19例患者伤口均Ⅰ期愈合,无切口感染、皮下积液等并发症.18例患者获得随访,随访3~30个月,无局部疼痛、牵拉等不适感,无复发病例.提示脱细胞真皮基质材料为未成年人、尚未婚育及有生育要求的男性腹股沟疝患者的治疗提供一种新的选择.

  20. Successful breast reconstruction using acellular dermal matrix can be recommended in healthy non-smoking patients

    DEFF Research Database (Denmark)

    Gunnarsson, Gudjon Leifur; Børsen-Koch, Mikkel; Arffmann, Susanne

    2013-01-01

    We present Scandinavia's first series of immediate alloplastic breast reconstructions with an acellular dermal matrix.......We present Scandinavia's first series of immediate alloplastic breast reconstructions with an acellular dermal matrix....

  1. Abdominal wall repair with human acellular dermal autograft

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    Roel E. Genders

    2011-12-01

    Full Text Available Repair of abdominal wall defects in the presence of contamination or infection is a significant problem. The loss of tissue warrants enforcement of the abdominal wall, preferably by autologous material. However, autologous repair often requires extensive surgery. This paper presents a review of available literature of placement of an acellular human dermis to repair an abdominal fascia defect, in contaminated as well as in non-contaminated surgical fields. It is illustrated with a case report that describes the successful reconstruction of an infected abdominal wall defect with a human acellular dermis allograft. A systematic literature review was undertaken with searches performed in the Pubmed and Cochrane databases for the period up till March 2009, using the search terms Alloderm [Substance Name], Hernia [Mesh] and the key words acellular dermis, acellular dermal matrix, human acellular dermal allograft and abdominal wall defect. To assess methodological quality, each article was subjected to a modification of the methodological index for non-randomized studies (MINORS according to Slim et al. Two items from the original index were not included because none of the studies selected had an unbiased assessment of the study end points and in none of the studies was a prospective calculation of the study size performed. Seventeen studies were included in the review. Data were extracted regarding study design, number of patients, surgical technique, followup period, contaminated or non-contaminated area of the fascia defect, mortality and morbidity (hemorrhage, seroma, wound dehiscence, infection of the operative procedure, the longterm results (removal of the graft, reherniation and bulging and level of evidencey. A total of 169 short-term complications and 151 longterm complications occurred after 643 surgical procedures reconstructing both contaminated and clean abdominal wall defects by implantation of an HADA. Human acellular dermal allograft

  2. Applications of acellular dermal matrix in revision breast reconstruction surgery.

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    Spear, Scott L; Sher, Sarah R; Al-Attar, Ali; Pittman, Troy

    2014-01-01

    Acellular dermal matrix has been used for over a decade in primary breast reconstruction. Few articles have specifically examined its use in revision breast reconstruction for fold malposition, capsular contracture, rippling, and symmastia. One hundred thirty-five revision breast reconstructive procedures using acellular dermal matrix (AlloDerm) in 118 patients (154 breasts) over a 5-year period were reviewed. Most procedures were revisions or part of the second stage of previous mastectomy reconstructions; three were revisions after reconstruction of congenital chest wall deformities. Fifty-seven revisions (37 percent) were for inferior fold malposition, followed by 40 (25.9 percent) for inferior pole support, 42 (27.2 percent) for capsular contracture, 10 (6.4 percent) for rippling, and five (3.2 percent) for symmastia. The overall complication rate was 5 percent. Revisions with acellular dermal matrix were successful in 147 of 154 breasts (95.5 percent). The most common complication was capsular contracture, occurring in five breasts (3.2 percent). There was one infection (0.6 percent), failure to lower the inframammary fold in one breast (0.6 percent), and one persistence of rippling (0.6 percent). The mean follow-up was 207 days. Acellular dermal matrix has proven to be a reliable tool for managing some of the most common and challenging problems in implant-based breast reconstruction. Although there are few published data on the success of more conventional solutions to fold malposition, lower pole support, and capsular contracture, the addition of acellular dermal matrix to buttress these repairs has been shown to provide a high likelihood of success with a low risk of complications.

  3. Acellular matrix of bovine pericardium bound with L-arginine

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    Kim, Hyo Joo [Department of Molecular Science and Technology, Ajou University, Suwon 443-749 (Korea, Republic of); Bae, Jin Woo [Department of Molecular Science and Technology, Ajou University, Suwon 443-749 (Korea, Republic of); Kim, Chun Ho [Laboratory of Tissue Engineering, Korea Cancer Center Hospital, Seoul 139-240 (Korea, Republic of); Lee, Jin Woo [Department of Orthopaedic Surgery, College of Medicine, Yonsei University, Seoul 120-749 (Korea, Republic of); Shin, Jung Woog [Department of Biomedical Engineering, Inje University, Gimhae 621-749 (Korea, Republic of); Park, Ki Dong [Department of Molecular Science and Technology, Ajou University, Suwon 443-749 (Korea, Republic of)

    2007-09-15

    Surface immobilization of bioactive molecules onto natural tissues has been interestingly studied for the development of new functional matrices for the replacement of lost or malfunctioning tissues. In this study, an acellular matrix of bovine pericardium (ABP) was chemically modified by the direct coupling of L-arginine after glutaraldehyde (GA) cross-linking. The effects of L-arginine coupling on durability and calcification were investigated and the biocompatibility was evaluated in vitro and in vivo. A four-step detergent and enzymatic extraction process has been utilized to remove cellular components from fresh bovine pericardium (BP). Microscopic observation confirmed that nearly all cellular constituents are removed. Thermal and mechanical properties showed that the durability of L-arginine-treated matrices increased as compared with control ABP and GA-treated ABP. Resistance to collagenase digestion revealed that modified matrices have greater resistance to enzyme digestion than control ABP and GA-treated ABP. The in vivo calcification study demonstrated much less calcium deposition on L-arginine-treated ABP than GA-treated one. In vitro cell viability results showed that ABP modified with L-arginine leads to a significant increase in attachment of human dermal fibroblasts. The obtained results attest to the usefulness of L-arginine-treated ABP matrices for cardiovascular bioprostheses.

  4. Porosity of porcine bladder acellular matrix: impact of ACM thickness.

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    Farhat, Walid; Chen, Jun; Erdeljan, Petar; Shemtov, Oren; Courtman, David; Khoury, Antoine; Yeger, Herman

    2003-12-01

    The objectives of this study are to examine the porosity of bladder acellular matrix (ACM) using deionized (DI) water as the model fluid and dextran as the indicator macromolecule, and to correlate the porosity to the ACM thickness. Porcine urinary bladders from pigs weighing 20-50 kg were sequentially extracted in detergent containing solutions, and to modify the ACM thickness, stretched bladders were acellularized in the same manner. Luminal and abluminal ACM specimens were subjected to fixed static DI water pressure (10 cm); and water passing through the specimens was collected at specific time interval. While for the macromolecule porosity testing, the diffusion rate and direction of 10,000 MW fluoroescein-labeled dextrans across the ACM specimens mounted in Ussing's chambers were measured. Both experiments were repeated on the thin stretched ACM. In both ACM types, the fluid porosity in both directions did not decrease with increased test duration (3 h); in addition, the abluminal surface was more porous to fluid than the luminal surface. On the other hand, when comparing thin to thick ACM, the porosity in either direction was higher in the thick ACM. Macromolecule porosity, as measured by absorbance, was higher for the abluminal thick ACM than the luminal side, but this characteristic was reversed in the thin ACM. Comparing thin to thick ACM, the luminal side in the thin ACM was more porous to dextran than in the thick ACM, but this characteristic was reversed for the abluminal side. The porcine bladder ACM possesses directional porosity and acellularizing stretched urinary bladders may increase structural density and alter fluid and macromolecule porosity.

  5. Delayed repair in a case of forearm fascial muscle herniation using non-cross-linked acellular porcine dermal matrix.

    Science.gov (United States)

    Hartmann, Christoph E A; Branford, Olivier A; Floyd, David

    2012-09-01

    The options for treatment of symptomatic muscle herniation in the limbs traditionally include fasciotomy, direct repair, tendon weave graft (palmaris longus), fascial graft (tensor fascia lata), and synthetic mesh (prolene). A recent case report has described the use of acellular cadaveric dermal matrix to reconstruct fascial defects in 2 cases. We describe the use of Strattice, a non-cross-linked acellular porcine dermal matrix, as a fascial underlay graft in a case of symptomatic upper limb muscle herniation. We propose that Strattice has the advantages over cadaveric dermal matrices in terms of avoiding the use of human donor tissue. It has suitable tensile properties to be used for reconstructing fascial defects.

  6. Progress in various crosslinking modification for acellular matrix

    Institute of Scientific and Technical Information of China (English)

    Yang Haitang; Tan Qiang; Zhao Heng

    2014-01-01

    Objective To review the current crosslinking strategies for acelluar matrix scaffold,laying the foundation for subsequent experiment.Data sources Data were mainly obtained from recent papers published in PubMed or indexed by Web of Science,with keyword like crosslinking.Results Various crosslinking strategies,including chemical,physical and biological methods,have been introduced to facilitate the performance of fresh acellular matrix.Chemical crosslinking reagents,involved in synthetic and naturally derived agents,need to be eliminated before implantation in case of their potential biotoxicity,although several crosslinking agents with less toxicity and specific characteristics have been developed.Physical crosslinking methods present to be safe,additive-free and relatively controllable for rapid surface functionalization with no consideration of remaining radioactivity.Biological crosslinking strategies have attracted great interest,and have been demonstrated to enhance collagen-based crosslinking since their preparations do not need toxic or potentially biologically contaminated substances and can be carried out under physiological conditions.Conclusions Kinds of crosslinking methods with its potential advantages have been developed to modify raw acelluar matrix,of which the performance are promising after being crosslinked by several crosslinking treatments.Further preclinical and clinical evaluations should be taken to vertify their safety and efficacy for the tissues and organs substitutes in tissue and regenerative medicine.

  7. Reconstruction of the abdominal wall by using a combination of the human acellular dermal matrix implant and an interpositional omentum flap after extensive tumor resection in patients with abdominal wall neoplasm:A preliminary result

    Institute of Scientific and Technical Information of China (English)

    Yan Gu; Rui Tang; Ding-Quan Gong; Yun-Liang Qian

    2008-01-01

    AIM:To present our trial using a combination of the human acellular dermal matrix (HADM) implant and an interpositional omentum flap to repair giant abdominal wall defects after extensive tumor resection.METHODS:Between February and October of 2007,three patients with giant defects of the abdominal wall after extensive tumor resection underwent reconstruction with a combination of HADM and omentum flap.Postoperative morbidities and signs of herniation were monitored.RESULTS:The abdominal wall reconstruction was successful in these three patients,there was no severe morbidity and no signs of herniation in the follow-up period.CONCLUSION:The combination of HADM and omentum flap offers a new,safe and effective alternative to traditional forms in the repair of giant abdominal wall defects.Further analysis of the long-term outcome and more cases are needed to assess the reliability of this technique.

  8. Porcine bladder acellular matrix (ACM): protein expression, mechanical properties.

    Science.gov (United States)

    Farhat, Walid A; Chen, Jun; Haig, Jennifer; Antoon, Roula; Litman, Jessica; Sherman, Christopher; Derwin, Kathleen; Yeger, Herman

    2008-06-01

    Experimentally, porcine bladder acellular matrix (ACM) that mimics extracellular matrix has excellent potential as a bladder substitute. Herein we investigated the spatial localization and expression of different key cellular and extracellular proteins in the ACM; furthermore, we evaluated the inherent mechanical properties of the resultant ACM prior to implantation. Using a proprietary decellularization method, the DNA contents in both ACM and normal bladder were measured; in addition we used immunohistochemistry and western blots to quantify and localize the different cellular and extracellular components, and finally the mechanical testing was performed using a uniaxial mechanical testing machine. The mean DNA content in the ACM was significantly lower in the ACM compared to the bladder. Furthermore, the immunohistochemical and western blot analyses showed that collagen I and IV were preserved in the ACM, but possibly denatured collagen III in the ACM. Furthermore, elastin, laminin and fibronectin were mildly reduced in the ACM. Although the ACM did not exhibit nucleated cells, residual cellular components (actin, myosin, vimentin and others) were still present. There was, on the other hand, no significant difference in the mean stiffness between the ACM and the bladder. Although our decellularization method is effective in removing nuclear material from the bladder while maintaining its inherent mechanical properties, further work is mandatory to determine whether these residual DNA and cellular remnants would lead to any immune reaction, or if the mechanical properties of the ACM are preserved upon implantation and cellularization.

  9. Porcine bladder acellular matrix (ACM): protein expression, mechanical properties

    Energy Technology Data Exchange (ETDEWEB)

    Farhat, Walid A [Department of Surgery, Division of Urology, University of Toronto and Hospital for Sick Children, Toronto, ON M5G 1X8 (Canada); Chen Jun; Haig, Jennifer; Antoon, Roula; Litman, Jessica; Yeger, Herman [Department of Developmental and Stem Cell Biology, Research Institute, Hospital for Sick Children, Toronto, ON M5G 1X8 (Canada); Sherman, Christopher [Department of Anatomic Pathology, Sunnybrook and Women' s College Health Sciences Centre, Toronto, ON (Canada); Derwin, Kathleen [Department of Biomedical Engineering, Lerner Research Institute and Orthopaedic Research Center, Cleveland Clinic Foundation, Cleveland, OH (United States)], E-mail: walid.farhat@sickkids.ca

    2008-06-01

    Experimentally, porcine bladder acellular matrix (ACM) that mimics extracellular matrix has excellent potential as a bladder substitute. Herein we investigated the spatial localization and expression of different key cellular and extracellular proteins in the ACM; furthermore, we evaluated the inherent mechanical properties of the resultant ACM prior to implantation. Using a proprietary decellularization method, the DNA contents in both ACM and normal bladder were measured; in addition we used immunohistochemistry and western blots to quantify and localize the different cellular and extracellular components, and finally the mechanical testing was performed using a uniaxial mechanical testing machine. The mean DNA content in the ACM was significantly lower in the ACM compared to the bladder. Furthermore, the immunohistochemical and western blot analyses showed that collagen I and IV were preserved in the ACM, but possibly denatured collagen III in the ACM. Furthermore, elastin, laminin and fibronectin were mildly reduced in the ACM. Although the ACM did not exhibit nucleated cells, residual cellular components (actin, myosin, vimentin and others) were still present. There was, on the other hand, no significant difference in the mean stiffness between the ACM and the bladder. Although our decellularization method is effective in removing nuclear material from the bladder while maintaining its inherent mechanical properties, further work is mandatory to determine whether these residual DNA and cellular remnants would lead to any immune reaction, or if the mechanical properties of the ACM are preserved upon implantation and cellularization.

  10. Engineering 3D bio-artificial heart muscle: the acellular ventricular extracellular matrix model.

    Science.gov (United States)

    Patel, Nikita M; Tao, Ze-Wei; Mohamed, Mohamed A; Hogan, Matt K; Gutierrez, Laura; Birla, Ravi K

    2015-01-01

    Current therapies in left ventricular systolic dysfunction and end-stage heart failure include mechanical assist devices or transplant. The development of a tissue-engineered integrative platform would present a therapeutic option that overcomes the limitations associated with current treatment modalities. This study provides a foundation for the fabrication and preliminary viability of the acellular ventricular extracellular matrix (AVEM) model. Acellular ventricular extracellular matrix was fabricated by culturing 4 million rat neonatal cardiac cells around an excised acellular ventricular segment. Acellular ventricular extracellular matrix generated a maximum spontaneous contractile force of 388.3 μN and demonstrated a Frank-Starling relationship at varying pretensions. Histologic assessment displayed cell cohesion and adhesion within the AVEM as a result of passive cell seeding.

  11. Bovine versus Porcine Acellular Dermal Matrix: A Comparison of Mechanical Properties

    Directory of Open Access Journals (Sweden)

    David M. Adelman, MD, PhD, FACS

    2014-05-01

    Conclusions: Before implantation, BADM is inherently stronger than PADM at equivalent thicknesses and considerably stronger at increased thicknesses. These results corroborate clinical data from a previous study in which PADM was associated with a higher intraoperative device failure rate. Although numerous properties of acellular dermal matrix contribute to clinical outcomes, surgeons should consider initial mechanical strength properties when choosing acellular dermal matrices for load-bearing applications such as hernia repair.

  12. Management of gingival recession with acellular dermal matrix graft: A clinical study

    Directory of Open Access Journals (Sweden)

    V R Balaji

    2016-01-01

    Full Text Available Aims and Objectives: Obtaining root coverage has become an important part of periodontal therapy. The aims of this studyare to evaluate the clinical efficacy of acellular dermal matrix graft in the coverage of denuded roots and also to examine the change in the width of keratinized gingiva. Materials and Methods: A total of 20 sites with more than or equal to 2 mm of recession depth were taken into the study, for treatment with acellular dermal matrix graft. The clinical parameters such as recession depth, recession width, width of keratinized gingiva, probing pocket depth (PD, and clinical attachment level (CAL were measured at the baseline, 8th week, and at the end of the study (16th week. The defects were treated with a coronally positioned pedicle graft combined with acellular dermal matrix graft. Results: Out of 20 sites treated with acellular dermal matrix graft, seven sites showed complete root coverage (100%, and the mean root coverage obtained was 73.39%. There was a statistically significant reduction in recession depth, recession width, and probing PD. There was also a statistically significant increase in width of keratinized gingiva and also gain in CAL. The postoperative results were both clinically and statistically significant (P < 0.0001. Conclusion: The results of this study were esthetically acceptable to the patients and clinically acceptable in all cases. From this study, it may be concluded that acellular dermal matrix graft is an excellent substitute for autogenous graft in coverage of denuded roots.

  13. Differentiation of mesenchymal stem cells into neuronal cells on fetal bovine acellular dermal matrix as a tissue engineered nerve scaffold

    Institute of Scientific and Technical Information of China (English)

    Yuping Feng; Jiao Wang; Shixin Ling; Zhuo Li; Mingsheng Li; Qiongyi Li; Zongren Ma; Sijiu Yu

    2014-01-01

    The purpose of this study was to assess fetal bovine acellular dermal matrix as a scaffold for supporting the differentiation of bone marrow mesenchymal stem cells into neural cells fol-lowing induction with neural differentiation medium. We performed long-term, continuous observation of cell morphology, growth, differentiation, and neuronal development using several microscopy techniques in conjunction with immunohistochemistry. We examined speciifc neu-ronal proteins and Nissl bodies involved in the differentiation process in order to determine the neuronal differentiation of bone marrow mesenchymal stem cells. The results show that bone marrow mesenchymal stem cells that differentiate on fetal bovine acellular dermal matrix display neuronal morphology with unipolar and bi/multipolar neurite elongations that express neuro-nal-speciifc proteins, includingβIII tubulin. The bone marrow mesenchymal stem cells grown on fetal bovine acellular dermal matrix and induced for long periods of time with neural differen-tiation medium differentiated into a multilayered neural network-like structure with long nerve ifbers that was composed of several parallel microifbers and neuronal cells, forming a complete neural circuit with dendrite-dendrite to axon-dendrite to dendrite-axon synapses. In addition, growth cones with filopodia were observed using scanning electron microscopy. Paraffin sec-tioning showed differentiated bone marrow mesenchymal stem cells with the typical features of neuronal phenotype, such as a large, round nucleus and a cytoplasm full of Nissl bodies. The data suggest that the biological scaffold fetal bovine acellular dermal matrix is capable of supporting human bone marrow mesenchymal stem cell differentiation into functional neurons and the subsequent formation of tissue engineered nerve.

  14. Development and characterization of a full-thickness acellular porcine cornea matrix for tissue engineering.

    Science.gov (United States)

    Du, Liqun; Wu, Xinyi

    2011-07-01

    Our aim was to produce a natural, acellular matrix from porcine cornea for use as a scaffold in developing a tissue-engineered cornea replacement. Full-thickness, intact porcine corneas were decellularized by immersion in 0.5% (wt/vol) sodium dodecyl sulfate. The resulting acellular matrices were then characterized and examined specifically for completeness of the decellularization process. Histological analyses of decellularized corneal stromas showed that complete cell and α-Gal removal was achieved, while the major structural proteins including collagen type I and IV, laminin, and fibronectin were retained. DAPI staining did not detect any residual DNA within the matrix, and the DNA contents, which reflect the presence of cellular materials, were significantly diminished in the decellularized cornea. The collagen content of the decellularized cornea was well maintained compared with native tissues. Uniaxial tensile testing indicated that decellularization did not significantly compromise the ultimate tensile strength of the tissue (P > 0.05). In vitro cytotoxicity assays using rabbit corneal fibroblast cultures excluded the presence of soluble toxins in the biomaterial. In vivo implantation to rabbit interlamellar stromal pockets showed good biocompability. In summary, a full-thickness natural acellular matrix retaining the major structural components and strength of the cornea has been successfully developed. The matrix is biocompatible with cornea-derived cells and has potential for use in corneal transplantation and tissue-engineering applications. © 2011, Copyright the Authors. Artificial Organs © 2011, International Center for Artificial Organs and Transplantation and Wiley Periodicals, Inc.

  15. Acellular dermal matrix based nipple reconstruction: A modified technique

    Directory of Open Access Journals (Sweden)

    Raghavan Vidya

    2017-09-01

    Full Text Available Nipple areolar reconstruction (NAR has evolved with the advancement in breast reconstruction and can improve self-esteem and, consequently, patient satisfaction. Although a variety of reconstruction techniques have been described in the literature varying from nipple sharing, local flaps to alloplastic and allograft augmentation, over time, loss of nipple projection remains a major problem. Acellular dermal matrices (ADM have revolutionised breast reconstruction more recently. We discuss the use of ADM to act as a base plate and strut to give support to the base and offer nipple bulk and projection in a primary procedure of NAR with a local clover shaped dermal flap in 5 breasts (4 patients. We used 5-point Likert scales (1 = highly unsatisfied, 5 = highly satisfied to assess patient satisfaction. Median age was 46 years (range: 38–55 years. Nipple projection of 8 mm, 7 mm, and 7 mms were achieved in the unilateral cases and 6 mm in the bilateral case over a median 18 month period. All patients reported at least a 4 on the Likert scale. We had no post-operative complications. It seems that nipple areolar reconstruction [NAR] using ADM can achieve nipple projection which is considered aesthetically pleasing for patients.

  16. 利用脂肪脱细胞基质构建组织工程化脂肪的实验研究%A Study on Human Acellular Adipose Tissue Matrix in Construction of Tissue Engineered Fat

    Institute of Scientific and Technical Information of China (English)

    宋玫; 刘毅; 惠玲

    2016-01-01

    Objective To investigate the possibility of human acellular adipose tissue matrix ( ACAM) in con-struction of tissue-engineered fat. Methods The adipose tissues were harvested from discarded human adipose tissue in abdominal skin graft operation, and ACAM was obtained after treatment of repeated freeze-thaw, organic solvent extrac-tion and enzyme digestion. The decellularization effect and microstructure of matrix was examined using histological stai-ning and scanning electron microscopy. The third-generation human adipose derived stromal cells ( hADSCs) were la-beled with DiI, and were cultivated with ACAM. The complexes biocompatibility was detected, and then cell-scaffold complexes were subcutaneously transplanted in the back of Wistar rats. The adipogenic capacity in vivo was judged using general and fluorescence microscopy, wet-weight determination, histological detection and oil red O staining. Results The obtained ACAM consisted completely of extracellular matrix without any cell remains. General and Scanning electron microscopy images showed that the acellular matrix had porous structure and good cell compatibility. The new adipose tis-sues formed 8 weeks after transplantation in experiment and control groups. The wet-weight of transplants in the experi-ment group was significantly heavier than that in the control group (P<0. 01). HE and red oil O staining confirmed that the graft was mature adipose tissue, and DiI fluorescent staining proved that it was exogenous ADSCs. Conclusion Hu-man ACAM has good biocompatibility and biodegradability, and therefore it is suitable for cell proliferation and differenti-ation. ACAM scaffold combined with hADSCs may successfully form tissue-engineered adipose tissue in vivo. It can be used as an ideal method to construct tissue-engineered adipose tissues.%目的 探讨以人脂肪组织脱细胞基质( ACAM )作为支架材料构建组织工程化脂肪组织的可行性. 方法 取腹部取皮术后的剩余人脂肪组

  17. Randomized controlled trial of minimally invasive surgery using acellular dermal matrix for complex anorectal fistula

    Institute of Scientific and Technical Information of China (English)

    Ma-Mu-Ti-Jiang; A; ba-bai-ke-re; Er-Ha-Ti; Ai

    2010-01-01

    AIM: To compare the efficacy and safety of acellular dermal matrix (ADM) bioprosthetic material and endorectal advancement flap (ERAF) in treatment of complex anorectal fistula. METHODS: Ninety consecutive patients with complex anorectal fistulae admitted to Anorectal Surgical Department of First Affi liated Hospital, Xinjiang Medical University from March 2008 to July 2009, were enrolled in this study. Complex anorectal fistula was diagnosed following its clinical, radiographic, or endoscopic diagnostic cr...

  18. Tissue-engineered graft constructed by self-derived cells and heterogeneous acellular matrix

    Institute of Scientific and Technical Information of China (English)

    HUANG Hui-min; WU Shao-feng; REN Hong

    2006-01-01

    Background: Endothelial and smooth muscle cells were used as seeding cells and heterogeneous acellularized matrix was used as scaffold to construct the tissue-engineered graft. Methods: A 2 weeks piglet was selected as a donor of seeding cells. Two-centimetre length of common carotid artery was dissected. Endothelial cells and smooth muscle cells were harvested by trypsin and collagenase digestion respectively. The isolated cells were cultured and expanded using routine cell culture technique.An adult sheep was used as a donor of acellularized matrix. The thoracic aorta was harvested and processed by a multi-step decellularizing technique to remove the original cells and preserve the elastic and collagen fibers. The cultured smooth muscle cells and endothelial cells were then seeded to the acellularized matrix and incubated in vitro for another 2 weeks. The cell seeded graft was then transplanted to the cell-donated piglet to substitute part of the native pulmonary artery. Results: The cultured cells from piglet were characterized as endothelial cells by the presence of specific antigens vWF and CD31, and smooth muscle cells by the presence of specific antigen α-actin on the cell surface respectively with immunohistochemical technique. After decellularizing processing for the thoracic aorta from sheep, all the cellular components were extracted and elastic and collagen fibers kept their original morphology and structure. The maximal load of acellular matrix was decreased and 20% lower than that of untreated thoracic aorta, but the maximal tensions between them were not different statistically and they had similar load-tension curves. Three months after transplantation, the animal was sacrificed and the graft was removed for observation. The results showed that the inner surfaces of the graft were smooth, without thrombosis and calcification. Under microscopy, a great number of growing cells could be seen and elastic and collagen fibers were abundant. Conclusion

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

    Directory of Open Access Journals (Sweden)

    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.

  20. Xenogeneic acellular conjunctiva matrix as a scaffold of tissue-engineered corneal epithelium.

    Directory of Open Access Journals (Sweden)

    Haifeng Zhao

    Full Text Available Amniotic membrane-based tissue-engineered corneal epithelium has been widely used in the reconstruction of the ocular surface. However, it often degrades too early to ensure the success of the transplanted corneal epithelium when treating patients with severe ocular surface disorders. In the present study, we investigated the preparation of xenogeneic acellular conjunctiva matrix (aCM and evaluated its efficacy and safety as a scaffold of tissue-engineered corneal epithelium. Native porcine conjunctiva was decellularized with 0.1% sodium dodecyl sulfate (SDS for 12 h at 37°C and sterilized via γ-irradiation. Compared with native conjunctiva, more than 92% of the DNA was removed, and more than 90% of the extracellular matrix components (glycosaminoglycan and collagen remained after the decellularization treatment. Compared with denuded amniotic membrane (dAM, the aCM possessed favorable optical transmittance, tensile strength, stability and biocompatibility as well as stronger resistance to degradation both in vitro and in vivo. The corneal epithelial cells seeded on aCM formed a multilayered epithelial structure and endured longer than did those on dAM. The aCM-based tissue-engineered corneal epithelium was more effective in the reconstruction of the ocular surface in rabbits with limbal stem cell deficiency. These findings support the application of xenogeneic acellular conjunctiva matrix as a scaffold for reconstructing the ocular surface.

  1. Porcine vesical acellular matrix graft of tunica albuginea for penile reconstruction

    Institute of Scientific and Technical Information of China (English)

    Kwan-Joong Joo; Byung-Soo Kim; Jeong-Ho Han; Chang-Ju Kim; Chil-Hun Kwon; Heung-Jae Park

    2006-01-01

    Aim: To characterize the feasibility of the surgical replacement of the penile tunica albuginea (TA) and to evaluate the value of a porcine bladder acellular matrix (BAM) graft. Methods: Acellular matrices were constructed from pigs'bladders by cell lysis, and then examined by scanning electron microscopy (SEM). Expression levels of the mRNA of the vascular endothelial growth factor (VEGF) receptor, fibroblast growth factor (FGF)-1 receptor, neuregulin, and brain-derived neurotrophic factor (BDNF) in the acellular matrix and submucosa of the pigs' bladders were determined through the reverse transcription-polymerase chain reaction (PCR). A 5 mm × 5 mm square was excised from the penile TA of nine rabbits. The defective TA was then covered in porcine BAM. Equal numbers of animals were sacrificed and histochemically examined at 2, 4 and 6 months after implantation. Results: SEM of the BAM showed collagen fibers with many pores. VEGF receptor, FGF-1 receptor and neuregulin mRNA were expressed in the porcine BAM; BDNF mRNA was not detected. Two months after implantation, the graft sites exhibited excellent healing without contracture, and the fusion between the graft and the neighboring normal TA appeared to be well established. There were no significant histological differences between the implanted tunica and the normal control tunica at 6 months after implantation. Conclusion: The porcine BAM graft resulted in a structure which was sufficiently like that of the normal TA. This implantation might be considered applicable to the reconstruction of the TA in conditions such as trauma or Peyronie's disease.

  2. The acellular matrix (ACM) for bladder tissue engineering: A quantitative magnetic resonance imaging study.

    Science.gov (United States)

    Cheng, Hai-Ling Margaret; Loai, Yasir; Beaumont, Marine; Farhat, Walid A

    2010-08-01

    Bladder acellular matrices (ACMs) derived from natural tissue are gaining increasing attention for their role in tissue engineering and regeneration. Unlike conventional scaffolds based on biodegradable polymers or gels, ACMs possess native biomechanical and many acquired biologic properties. Efforts to optimize ACM-based scaffolds are ongoing and would be greatly assisted by a noninvasive means to characterize scaffold properties and monitor interaction with cells. MRI is well suited to this role, but research with MRI for scaffold characterization has been limited. This study presents initial results from quantitative MRI measurements for bladder ACM characterization and investigates the effects of incorporating hyaluronic acid, a natural biomaterial useful in tissue-engineering and regeneration. Measured MR relaxation times (T(1), T(2)) and diffusion coefficient were consistent with increased water uptake and glycosaminoglycan content observed on biochemistry in hyaluronic acid ACMs. Multicomponent MRI provided greater specificity, with diffusion data showing an acellular environment and T(2) components distinguishing the separate effects of increased glycosaminoglycans and hydration. These results suggest that quantitative MRI may provide useful information on matrix composition and structure, which is valuable in guiding further development using bladder ACMs for organ regeneration and in strategies involving the use of hyaluronic acid.

  3. Treatment of amalgam tattoo with a subepithelial connective tissue graft and acellular dermal matrix.

    Science.gov (United States)

    Thumbigere-Math, Vivek; Johnson, Deborah K

    2014-04-01

    A 54-year-old female was referred for management of a large amalgam tattoo involving the alveolar mucosa between teeth #6 and #9. The lesion had been present for over 20 years following endodontic treatment of teeth #7 and #8. A two-stage surgical approach was used to remove the pigmentation, beginning with removal of amalgam fragments from the underlying bone and placement of a subepithelial connective tissue graft and acellular dermal matrix to increase soft tissue thickness subadjacent to the amalgam. Following 7 weeks of healing, gingivoplasty was performed to remove the overlying pigmented tissue. At the 21-month follow-up appointment, the patient exhibited naturally appearing soft tissue with no evidence of amalgam tattoo.

  4. A novel porcine acellular dermal matrix scaffold used in periodontal regeneration

    Institute of Scientific and Technical Information of China (English)

    Jing Guo; Hui Chen; Ying Wang; Cheng-Bo Cao; Guo-Qiang Guan

    2013-01-01

    Regeneration of periodontal tissue is the most promising method for restoring periodontal structures. To find a suitable bioactive three- dimensional scaffold promoting cell proliferation and differentiation is critical in periodontal tissue engineering. The objective of this study was to evaluate the biocompatibility of a novel porcine acellular dermal matrix as periodontal tissue scaffolds both in vitroand in vivo. The scaffolds in this study were purified porcine acellular dermal matrix (PADM) and hydroxyapatite-treated PADM (HA-PADM). The biodegradation patterns of the scaffolds were evaluated in vitro. The biocompatibility of the scaffolds in vivo was assessed by implanting them into the sacrospinal muscle of 20 New Zealand white rabbits. The hPDL cells were cultured with PADM or HA-PADM scaffolds for 3, 7, 14, 21 and 28 days. Cell viability assay, scanning electron microscopy (SEM), hematoxylin and eosin (H&E) staining, immunohistochemistry and confocal microscopy were used to evaluate the biocompatibility of the scaffolds. In vitro, both PADM and HA-PADM scaffolds displayed appropriate biodegradation pattern, and also, demonstrated favorable tissue compatibility without tissue necrosis, fibrosis and other abnormal response. The absorbance readings of the WST-1 assay were increased with the time course, suggesting the cell proliferation in the scaffolds. The hPDL cells attaching, spreading and morphology on the surface of the scaffold were visualized by SEM, H&E staining, immnuohistochemistry and confocal microscopy, demonstrated that hPDL cells were able to grow into the HA-PADM scaffolds and the amount of cells were growing up in the course of time. This study proved that HA-PADM scaffold had good biocompatibility in animals in vivoand appropriate biodegrading characteristics in vitro. The hPDL cells were able to proliferate and migrate into the scaffold. These observations may suggest that HA-PADM scaffold is a potential cell carrier

  5. Experimental total wrapping of breast implants with acellular dermal matrix: a preventive tool against capsular contracture in breast surgery?

    Science.gov (United States)

    Schmitz, Marweh; Bertram, Martin; Kneser, Ulrich; Keller, Andrea K; Horch, Raymund E

    2013-10-01

    Capsular contracture remains a hitherto unsolved complication after implantation of silicone gel-filled breast prostheses. Based on clinical and experimental data, the use of an acellular dermal matrix as a sheath around implants may lead to lesser capsular contracture acting as a proposed biological environment mimicking wound bed tissue. The aim of our study was to analyse the tissue reaction after implantation of silicone prosthesis with and without an envelope of acellular dermal matrix. Implantation of 60 silicone prostheses in the back of Lewis rats were carried out, randomly paired taking one rat from group A and one from group B. Group A included implants completely enveloped with xenogenic acellular dermis and group B undraped silicone implants. At 3, 6 and 12 weeks postoperatively, the samples were explanted and subjected to histological and immunohistochemical evaluation. A new myofibroblast tissue layer was identified in proximity to the implant in both groups. The thickness of the layer in group A was continuously thinner than in group B regarding the different explantation time points. Implants completely wrapped with acellular dermal matrix showed significantly lesser inflammatory signs at 3 and 12 weeks after implantation compared to controls. Cell proliferation after 12 weeks was significantly decreased in group A. The slight myofibroblast layer and reduced rate of inflammation and proliferation in the treatment group show a positive effect of total acellular dermal matrix envelope and hypothesise the decrease of capsular contracture in long-term periods. Copyright © 2013 British Association of Plastic, Reconstructive and Aesthetic Surgeons. Published by Elsevier Ltd. All rights reserved.

  6. Results of Acellular Dermis Matrix Graft Used for Tympanoplasty in Guinea Pig Model

    Directory of Open Access Journals (Sweden)

    Farhad Farahani

    2015-03-01

    Full Text Available Introduction: To describe the underlay tympanoplasty technique using an acellular dermal graft(AlloDerm for tympanic membrane (TM reconstruction in a guinea pig model and to demonstrate the feasibility of the technique using AlloDerm tissue harvested from the prepuce as a source of tissue for future grafting in human TM reconstruction.   Materials and Methods: The prepuce was divided during circumcision and the acellular dermis was prepared using a number of standard processes. Two groups of guinea pigs were prepared. In the case group (20 guinea pigs and 40 ears removal of TM was performed with tympanoplasty using AlloDerm, and in the control group (eight guinea pigs and 16 ears, removal of TM was performed without tympanoplasty. In each group, the TM was completely removed in one ear and partially removed on the other side, and the integrity of the TMs was re-evaluated after 8 weeks.   Results: In the case group, the healing rates in the completely and partially removed TMs were 83.3% and 94.4%, respectively. The difference in healing rate (0% and 66.7%, respectively was statistically significant (P

  7. Biopolymer gel matrix as acellular scaffold for enhanced dermal tissue regeneration.

    Science.gov (United States)

    Judith, Rangasamy; Nithya, Mariappan; Rose, Chellan; Mandal, Asit Baran

    2012-07-01

    Biological grafts have drawbacks such as donor scarcity, disease transmission, tissue infection, while the scaffolds of either collagen or chitosan fabrics fail to become part of the tissue at the wound site, though they favor the formation of connective tissue matrix. This study developed a novel composite consisting of the combination of atelocollagen and chitosan in order to provide a biodegradable molecular matrix in gel form as a biomimetic surface for cell attachment, to promote the wound healing in excision wounds. We found that the topical application of biopolymer composite on the wound promoted cell proliferation, migration and collagen deposition overtime. The enhanced cellular activity in the collagen-chitosan treated wound tissue was also assed by increased levels of Platelet derived growth factor (PDGF) and Nerve growth factor (NGF) associated with elevated levels of antioxidants and decreased level of lipid peroxidation. The acellular matrix-like topical application material is designed to guide the eventual re-establishment of an anatomically normal skin. The results of this study demonstrate the feasibility of multi-cell regeneration on a molecular system that mimics tissue engineering in vivo.

  8. 人肺癌H460细胞在脱细胞化鼠肺基质支架中的生长%Human lung cancer cells grow on acellular rat lung matrix

    Institute of Scientific and Technical Information of China (English)

    朱海波; 张倩; 王秀丽; 徐小玉; 王宏; 王玲

    2014-01-01

    Objective To explore the feasibility of human lung canccr cells grown in a decellularized rat lung matrix by perfusion.Methods Lungs were harvested from adult SD rats.Native cells of rat lungs were removed using 0.1% sodium dodecyl sulfate (SDS) and 1% Triton X-100 by perfusion to create a decellularized rat lung matrix.After decellularization,Human lung cancer H460 cells were implauted into the decellularized rat lung matrix and grown in a customized bioreactor with perfusion of oxygenated media for 1-2 weeks.Results Decellularized rat lung matrix showed preservation of matrix architecture devoid of all rat cells.H460 cells could grow in the bioreactor.Conclusions Human lung cancer H460 cells can grow in a customized bioreactor on a decellularized rat lung matrix.This ex vivo model can be used potentially to gain a deeper understanding of the biologic processes involved in human lung cancer.%目的 探索人肺癌细胞在用灌注法脱细胞的鼠肺基质支架中生长的可行性.方法 取成年SD大鼠心肺组织,利用0.1%十二烷基磺酸钠(SDS)溶液及1% TritonX-100溶液对离体的鼠肺行灌注法脱细胞,将人肺癌H460细胞株种植于脱细胞化鼠肺基质中,并将其置于特制的生物反应器中灌注培养l~2周.结果 脱细胞化后的鼠肺基质去除了大鼠自身细胞,保留了细胞外基质结构.H460细胞在脱细胞化鼠肺基质支架中生长.结论 人肺癌H460细胞株能在去细胞化的鼠肺基质支架中生长,这种间接体内模型的成功建立对人类肺癌的生物学进展研究有重要的意义.

  9. Tissue performance of bladder following stretched electrospun silk fibroin matrix and bladder acellular matrix implantation in a rabbit model.

    Science.gov (United States)

    Huang, Jian-Wen; Xu, Yue-Min; Li, Zhao-Bo; Murphy, Sean V; Zhao, Weixin; Liu, Qiang-Qiang; Zhu, Wei-Dong; Fu, Qiang; Zhang, Yao-Peng; Song, Lu-Jie

    2016-01-01

    The goal of this study was to investigate the tissue performance of bladder following stretched electrospun silk fibroin matrix (SESFM) implantation compared with bladder acellular matrix (BAM). We compared SESFM with BAM based on porosity and pore size. Scaffolds were separately transplanted into opposite walls of the bladder of 30 rabbits after stripping the bladder mucosa and smooth muscle (1.5 × 2.0 cm(2)). Gross anatomical observation, histological analysis and muscle contractility studies were performed at 2, 4, and 8 weeks post-op. SESFM has higher porosity and larger pore size compared with BAM (p calculus was evident in 7/10 rabbits. Histological analysis showed that SESFM and BAM promoted similar degree of urothelium regeneration (p > 0.05). However, SESFM promoted a higher degree of smooth muscle and vessel regeneration compared to BAM (p < 0.05). In addition, muscle strips supported by SESFM displayed higher contractile responses to carbachol, KCl, and phenylephrine compared with BAM. At 8 weeks, both matrices elicited similar mild acute and chronic inflammatory reactions. Our results demonstrated that SESFM has greater ability to promote bladder tissue regeneration with structural and functional properties compared to BAM, and with similar biocompatibility. © 2015 Wiley Periodicals, Inc.

  10. Xenogeneic acellular dermal matrix in combination with pectoralis major myocutaneous flap reconstructs hypopharynx and cervical esophagus.

    Science.gov (United States)

    Yin, Danhui; Tang, Qinglai; Wang, Shuang; Li, Shisheng; He, Xiangbo; Liu, Jiajia; Liu, Bingbing; Yang, Mi; Yang, Xinming

    2015-11-01

    The aim of this study was to explore xenogeneic acellular dermal matrix (ADM) in combination with pectoralis major myocutaneous flap in hypopharynx and cervical esophagus reconstruction. A total of five patients were treated with this surgical method to reconstruct hypopharynx and cervical esophagus in Second Xiangya Hospital between January 2012 and April 2013. Four of them had hypopharyngeal carcinoma with laryngeal and cervical esophageal invasion, while the fifth patient with hypopharyngeal cancer had developed scars and atresia after postoperative radiotherapy. The defect length after hypopharyngeal and cervical esophageal resection was 6-8 cm, and was repaired by a combination of ADM and pectoralis major myocutaneous flap by our team. Interestingly, the four patients had primary healing and regained their eating function about 2-3 weeks after surgery, the fifth individual suffered from pharyngeal fistula, but recovered after dressing change about 2 months. Postoperative esophageal barium meals revealed that the pharynx and esophagus were unobstructed in all five patients. Xenogeneic ADM in combination with pectoralis major myocutaneous flap for hypopharynx and cervical esophagus reconstruction is a simple, safe and effective method with fewer complications. Nevertheless, according to the defect length of the cervical esophagus, the patients need to strictly follow the medical advice.

  11. A anorectal fistula treatment with acellular extracellular matrix: A new technique

    Institute of Scientific and Technical Information of China (English)

    Wei-Liang Song; Zhen-Jun Wang; Yi Zheng; Xin-Qing Yang; Ya-Ping Peng

    2008-01-01

    AIM:To investigate a new technique of the anorectal fistula treatment with acellular extracellular matrix (AEM).METHODS: Thirty patients with anorectal fistula were treated with AEM.All fistula tracts and primary openings were identified using conventional fistula probe.All tracts were curetted with curet and irrigated with hydrogen peroxide and metronidazole.The AEM was pulled into the fistula tract from secondary to primary opening.The material was secured at the level of the primary opening.The excess AEM was trimmed at skin level at the secondary opening.RESULTS: All of the 30 patients had successful closure of their fistula after a 7-14 d follow-up.The healing rate of anal fistula in treatment group was 100%.The ache time,healing time and anal deformation of treatment group were obviously superior to traditional surgical methods.CONCLUSION: Using AEM anal fistula plug in treatment that causes the anorectal fistula is safe and successful in 100% of patients.It can reduce pain,shorten disease course and protect anal function.

  12. A anorectal fistula treatment with acellular extracellular matrix: A new technique

    Science.gov (United States)

    Song, Wei-Liang; Wang, Zhen-Jun; Zheng, Yi; Yang, Xin-Qing; Peng, Ya-Ping

    2008-01-01

    AIM: To investigate a new technique of the anorectal fistula treatment with acellular extracellular matrix (AEM). METHODS: Thirty patients with anorectal fistula were treated with AEM. All fistula tracts and primary openings were identified using conventional fistula probe. All tracts were curetted with curet and irrigated with hydrogen peroxide and metronidazole. The AEM was pulled into the fistula tract from secondary to primary opening. The material was secured at the level of the primary opening. The excess AEM was trimmed at skin level at the secondary opening. RESULTS: All of the 30 patients had successful closure of their fistula after a 7-14 d follow-up. The healing rate of anal fistula in treatment group was 100%. The ache time, healing time and anal deformation of treatment group were obviously superior to traditional surgical methods. CONCLUSION: Using AEM anal fistula plug in treatment that causes the anorectal fistula is safe and successful in 100% of patients. It can reduce pain, shorten disease course and protect anal function. PMID:18720541

  13. Biological function evaluation and effects of laser micro-pore burn-denatured acellular dermal matrix.

    Science.gov (United States)

    Zhang, Youlai; Zeng, Yuanlin; Xin, Guohua; Zou, Lijin; Ding, Yuewei; Duyin, Jiang

    2017-08-18

    In the field of burns repairs, many problems exist in the shortage of donor skin, the expense of allograft or xenograft skin, temporary substitution and unsatisfactory extremity function after wound healing. Previous studies showed that burn-denatured skin could return to normal dermis formation and function. This study investigates the application of laser micro-pore burn-denatured acellular dermis matrix (DADM) from an escharotomy in the repair of burn wounds and evaluates the biological properties and wound repair effects of DADM in implantation experiments in Kunming mice. Specific-pathogen-free (SPF) Kunming mice were used in this study. A deep II° burn wound was created on the dorsum of the mice by an electric heated water bath. The full-thickness wound tissue was harvested. The necrotic tissue and subcutaneous tissue were removed. The denatured dermis was preserved and treated with 0.25% trypsin, 0.5% Triton X-100. The DADM was drilled by laser micro-pore. The biological properties and grafting effects of laser micro-pore burn-DADM were evaluated by morphology, cytokine expression levels and subcutaneous implantation experiments in Kunming mice. We found statistical significance (Plaser micro-pore burn-DADM (experimental group) compared to the control group (no laser micro-pore burn-DADM). Cytokine expression level was different in the dermal matrixes harvested at various time points after burn (24h, 48h, 72h and infected wound group). Comparing the dermal matrix from 24h burn tissue to infected wound tissue, the expression level of IL-6, MMP-24, VE-cadherin and VEGF were decreased. We found no inflammatory cells infiltration in the dermal matrix were observed in both experimental and control groups (24h burn group), while the obviously vascular infiltration and fiber fusion were observed in the experimental group after subcutaneous implantation experiments. There was better bio-performance, low immunogenicity and better dermal incorporation after treated

  14. Flexor tendon tissue engineering: acellularization of human flexor tendons with preservation of biomechanical properties and biocompatibility.

    Science.gov (United States)

    Pridgen, Brian C; Woon, Colin Y L; Kim, Maxwell; Thorfinn, Johan; Lindsey, Derek; Pham, Hung; Chang, James

    2011-08-01

    Acellular human tendons are a candidate scaffold for tissue engineering flexor tendons of the hand. This study compared acellularization methods and their compatibility with allogeneic human cells. Human flexor tendons were pretreated with 0.1% ethylenediaminetetracetic acid (EDTA) for 4  h followed by 24  h treatments of 1% Triton X-100, 1% tri(n-butyl)phosphate, or 0.1% or 1% sodium dodecyl sulfate (SDS) in 0.1% EDTA. Outcomes were assessed histologically by hematoxylin and eosin and SYTO green fluorescent nucleic acid stains and biochemically by a QIAGEN DNeasy kit, Sircol collagen assay, and 1,9 dimethylmethylene blue glycosaminoglycan assay. Mechanical data were collected using a Materials Testing System to pull to failure tendons acellularized with 0.1% SDS. Acellularized tendons were re-seeded in a suspension of human dermal fibroblasts. Attachment of viable cells to acellularized tendon was assessed biochemically by a cell viability assay and histologically by a live/dead stain. Data are reported as mean±standard deviation. Compared with the DNA content of fresh tendons (551±212  ng DNA/mg tendon), only SDS treatments significantly decreased DNA content (1% SDS [202.8±37.4  ng DNA/mg dry weight tendon]; 0.1% SDS [189±104  ng DNA/mg tendon]). These findings were confirmed by histology. There was no decrease in glycosaminoglycans or collagen following acellularization with SDS. There was no difference in the ultimate tensile stress (55.3±19.2 [fresh] vs. 51.5±6.9 [0.1% SDS] MPa). Re-seeded tendons demonstrated attachment of viable cells to the tendon surface using a viability assay and histology. Human flexor tendons were acellularized with 0.1% SDS in 0.1% EDTA for 24  h with preservation of mechanical properties. Preservation of collagen and glycoaminoglycans and re-seeding with human cells suggest that this scaffold is biocompatible. This will provide a promising scaffold for future human flexor tendon tissue engineering studies to

  15. Daily Serum Collection after Acellular Dermal Matrix-Assisted Breast Reconstruction

    Directory of Open Access Journals (Sweden)

    Glenda Giorgia Caputo

    2015-05-01

    Full Text Available BackgroundThe acellular dermal matrix (ADM-assisted breast reconstruction technique is widely known, but discouraging results due to early postoperative complications have been reported. As the literature identifies seroma as the most common issue after breast surgery without identifying its pathogenesis, we aimed to report the trend of postoperative daily serum collection after ADM-assisted breast reconstruction and compare it with data in the literature in order to discover more about this little-known topic.MethodsA retrospective study on 28 consecutive patients who received ADM-assisted breast reconstruction between February 2013 and February 2014 was performed. In order to reduce the number of variables that could affect serum production, only one brand of ADM was used and all tissues were handled gently and precisely. The daily drainage volume was recorded per patient during the first four days of hospitalization. Likewise, postoperative complications were noted during routine follow-up.ResultsIn total, five (17.9% bilateral and 23 (82.1% unilateral ADM-assisted breast reconstructions (33 implants were performed. The mean age, body mass index, and length of hospital stay were 53.6 years, 21.3 kg/m2, and 4.5 days, respectively. One major complication led to implant loss (3.0%, and nine minor complications were successfully treated with ambulatory surgery (27.3%. Serum collection linearly decreased after 24 hours postoperatively.ConclusionsDaily drainage decreased following the theoretical decline of acute inflammation. In concordance with the literature, daily serum production may not be related to the use of ADM.

  16. Subcutaneous Implant-based Breast Reconstruction with Acellular Dermal Matrix/Mesh: A Systematic Review.

    Science.gov (United States)

    Salibian, Ara A; Frey, Jordan D; Choi, Mihye; Karp, Nolan S

    2016-11-01

    The availability of acellular dermal matrix (ADM) and synthetic mesh products has prompted plastic surgeons to revisit subcutaneous implant-based breast reconstruction. The literature is limited, however, with regards to evidence on patient selection, techniques, and outcomes. A systematic review of the Medline and Cochrane databases was performed for original studies reporting breast reconstruction with ADM or mesh, and subcutaneous implant placement. Studies were analyzed for level of evidence, inclusion/exclusion criteria for subcutaneous reconstruction, reconstruction characteristics, and outcomes. Six studies (186 reconstructions) were identified for review. The majority of studies (66.7%) were level IV evidence case series. Eighty percent of studies had contraindications for subcutaneous reconstruction, most commonly preoperative radiation, high body mass index, and active smoking. Forty percent of studies commenting on patient selection assessed mastectomy flap perfusion for subcutaneous reconstruction. Forty-five percent of reconstructions were direct-to-implant, 33.3% 2-stage, and 21.5% single-stage adjustable implant, with ADM utilized in 60.2% of reconstructions versus mesh. Pooled complication rates included: major infection 1.2%, seroma 2.9%, hematoma 2.3%, full nipple-areola complex necrosis 1.1%, partial nipple-areola complex necrosis 4.5%, major flap necrosis 1.8%, wound healing complication 2.3%, explantation 4.1%, and grade III/IV capsular contracture 1.2%. Pooled short-term complication rates in subcutaneous alloplastic breast reconstruction with ADM or mesh are low in preliminary studies with selective patient populations, though techniques and outcomes are variable across studies. Larger comparative studies and better-defined selection criteria and outcomes reporting are needed to develop appropriate indications for performing subcutaneous implant-based reconstruction.

  17. Plastic Surgery and Acellular Dermal Matrix: Highlighting Trends from 1999 to 2013.

    Science.gov (United States)

    Daar, David A; Gandy, Jessica R; Clark, Emily G; Mowlds, Donald S; Paydar, Keyianoosh Z; Wirth, Garrett A

    2016-05-01

    The last decade has ushered in a rapidly expanding global discussion regarding acellular dermal matrix (ADM) applications, economic analyses, technical considerations, benefits, and risks, with recent emphasis on ADM use in breast surgery. This study aims to evaluate global trends in ADM research using bibliometric analysis. The top nine Plastic Surgery journals were determined by impact factor (IF). Each issue of the nine journals between 1999 and 2013 was accessed to compile a database of articles discussing ADM. Publications were further classified by IF, authors' geographic location, study design, and level of evidence (LOE, I-V). Productivity index and productivity share were calculated for each region. In total, 256 ADM articles were accessed. The annual global publication volume increased significantly by 4.2 (0.87) articles per year (p<0.001), with a mean productivity index of 36.3 (59.0). The mean impact factor of the nine journals increased significantly from 0.61 (0.11) to 2.47 (0.99) from 1993 to 2013 (p<0.001). Despite this increase in the global ADM literature, the majority of research was of weaker LOE (level I: 2.29% and level II: 9.17%). USA contributed the most research (87%), followed by Asia (4.76%) and Western Europe (4.71%). USA contributed the greatest volume of research. Regarding clinical application of ADM, the majority of publications focused on ADM use in breast surgery, specifically breast reconstruction (154 articles, 60.2%). The majority of research was of lower LOE; thus, efforts should be made to strengthen the body of literature, particularly with regard to cost analysis.

  18. Using porcine acellular collagen matrix (Pelvicol® in bladder augmentation: experimental study

    Directory of Open Access Journals (Sweden)

    Ayyildiz Ali

    2006-01-01

    Full Text Available PURPOSE: Evaluate the rabbit augmented bladder with PelvicolÒ. MATERIALS AND METHODS: Twenty New Zealand rabbits were divided into 4 groups. Bladder augmentation was performed using a 10 x 10 mm sized porcine acellular collagen matrix. The material was placed on the dome of the bladder wall as a patch with 5-0 polyglycolic sutures. The bladder was resected on the 7th, 14th day, 30th and 90th days, and processed for histological analysis. RESULTS: No stone formation was found in the first, second and fourth weeks. In the first week, there was inflammatory appearance and roughness in the reconstructed area when compared to other sites on the bladder wall. The material could not be seen in some bladders because of acute inflammatory reaction. The normal bladder epithelium was found on the part of the bladder wall that follows the surface of the eroded material. In the second week, edema was observed through the bladder wall. Perivesical fat tissue increased and it was not easy to distinguish it from the surrounding area. In the fourth week, the bladder wall was thickened and there was a sensation of hardness present. The inner and outer surface of the material was darker than in the other bladders. In the third month, there was no inflammatory reaction; however, there was micro calcification and irregular detrusor regeneration. CONCLUSIONS: PelvicolÒ cannot be suitable material for bladder augmentation because of the resultant micro calcification, thickening of the bladder wall and irregular development of detrusor regeneration.

  19. Alternatives to Acellular Dermal Matrix: Utilization of a Gore DualMesh Sling as a Cost-Conscious Adjunct for Breast Reconstruction.

    Science.gov (United States)

    Grow, Jacob N; Butterworth, James; Petty, Paul

    2017-01-01

    Objective: This study seeks an alternative to acellular dermal matrix in 2-staged breast reconstruction while minimizing cost. It was hypothesized that use of a Gore DualMesh would allow for similar intraoperative tissue expander fill volumes, time to second-stage reconstruction, and number of postoperative fills compared with acellular dermal matrix at only a fraction of the expense. Methods: Retrospective analysis comparing Gore DualMesh (59 breasts, 34 patients), acellular dermal matrix (13 breasts, 8 patients), and total muscle coverage (25 breasts, 14 patients) for postmastectomy breast reconstruction was performed. Time to second-stage reconstruction, number of expansions, and relative initial fill volumes were compared between the 3 groups. Secondarily, complication rates were also considered, including seroma, infection, expander/implant explantation, removal of mesh, and capsular contracture. Statistical analysis was performed utilizing the Fisher exact test and the χ(2) test for categorical variables and the Mann-Whitney U test for continuous variables. Results: Relative initial fill volumes, number of expansions, and time to second-stage reconstruction showed no statistical difference between the acellular dermal matrix and Gore DualMesh groups (P = .494, P = .146, and P = .539, respectively). Furthermore, the Gore DualMesh group underwent significantly fewer fills (P Gore DualMesh represents a safe alternative to acellular dermal matrix for breast reconstruction with similar aesthetic results in certain patients at a fraction of the cost.

  20. Three-dimensional scaffolds of acellular human and porcine lungs for high throughput studies of lung disease and regeneration.

    Science.gov (United States)

    Wagner, Darcy E; Bonenfant, Nicholas R; Sokocevic, Dino; DeSarno, Michael J; Borg, Zachary D; Parsons, Charles S; Brooks, Elice M; Platz, Joseph J; Khalpey, Zain I; Hoganson, David M; Deng, Bin; Lam, Ying W; Oldinski, Rachael A; Ashikaga, Takamaru; Weiss, Daniel J

    2014-03-01

    Acellular scaffolds from complex whole organs such as lung are being increasingly studied for ex vivo organ generation and for in vitro studies of cell-extracellular matrix interactions. We have established effective methods for efficient de and recellularization of large animal and human lungs including techniques which allow multiple small segments (∼ 1-3 cm(3)) to be excised that retain 3-dimensional lung structure. Coupled with the use of a synthetic pleural coating, cells can be selectively physiologically inoculated via preserved vascular and airway conduits. Inoculated segments can be further sliced for high throughput studies. Further, we demonstrate thermography as a powerful noninvasive technique for monitoring perfusion decellularization and for evaluating preservation of vascular and airway networks following human and porcine lung decellularization. Collectively, these techniques are a significant step forward as they allow high throughput in vitro studies from a single lung or lobe in a more biologically relevant, three-dimensional acellular scaffold. Copyright © 2013 Elsevier Ltd. All rights reserved.

  1. Sustained release of VEGF from PLGA nanoparticles embedded thermo-sensitive hydrogel in full-thickness porcine bladder acellular matrix

    Directory of Open Access Journals (Sweden)

    Song Hua

    2011-01-01

    Full Text Available Abstract We fabricated a novel vascular endothelial growth factor (VEGF-loaded poly(lactic-co-glycolic acid (PLGA-nanoparticles (NPs-embedded thermo-sensitive hydrogel in porcine bladder acellular matrix allograft (BAMA system, which is designed for achieving a sustained release of VEGF protein, and embedding the protein carrier into the BAMA. We identified and optimized various formulations and process parameters to get the preferred particle size, entrapment, and polydispersibility of the VEGF-NPs, and incorporated the VEGF-NPs into the (poly(ethylene oxide-poly(propylene oxide-poly(ethylene oxide (Pluronic® F127 to achieve the preferred VEGF-NPs thermo-sensitive gel system. Then the thermal behavior of the system was proven by in vitro and in vivo study, and the kinetic-sustained release profile of the system embedded in porcine bladder acellular matrix was investigated. Results indicated that the bioactivity of the encapsulated VEGF released from the NPs was reserved, and the VEGF-NPs thermo-sensitive gel system can achieve sol-gel transmission successfully at appropriate temperature. Furthermore, the system can create a satisfactory tissue-compatible environment and an effective VEGF-sustained release approach. In conclusion, a novel VEGF-loaded PLGA NPs-embedded thermo-sensitive hydrogel in porcine BAMA system is successfully prepared, to provide a promising way for deficient bladder reconstruction therapy.

  2. Dermal fat graft from simultaneous abdominoplasty as an adjunct to revision aesthetic and reconstructive breast surgery: A poor man's acellular dermal matrix?

    Directory of Open Access Journals (Sweden)

    F. Xie

    2014-01-01

    CONCLUSION: We herein report the use of free dermal fat graft in revision aesthetic and reconstructive surgery in a manner akin to recent acellular dermal matrix use. The comparable enhanced aesthetic outcomes, minimal complication rate and substantial cost savings merit dissemination to a global audience and encourage surgeons to consider this economic alternative.

  3. The impact of acellular dermal matrix on tissue expander/implant loss in breast reconstruction: an analysis of the tracking outcomes and operations in plastic surgery database.

    Science.gov (United States)

    Pannucci, Christopher J; Antony, Anuja K; Wilkins, Edwin G

    2013-07-01

    Use of acellular dermal matrix in breast reconstruction has been associated with increased complications. However, existing studies are generally small, from single centers, and underpowered to control for confounding using regression techniques. Here, the Tracking Outcomes and Operations in Plastic Surgery database was used to examine the effect of acellular dermal matrix on expander/implant loss when controlling for other confounders. Analysis was limited to patients having tissue expander or implant-based breast reconstruction. Surgeon-reported data, International Classification of Diseases, Ninth Edition codes, and Current Procedural Terminology codes were used to identify independent variables. The dependent variable of interest was 30-day rates of tissue expander or implant loss. Bivariate statistics were performed. Multivariable logistic regression identified independent predictors of expander/implant loss when controlling for other confounders. Data were available for 14,249 patients. The overall rate of expander/implant loss was 2.05 percent. Bivariate analysis demonstrated acellular dermal matrix was associated with an absolute increase in expander/implant loss of 0.7 percent (1.88 percent versus 2.58 percent, p = 0.012). The regression model demonstrated that rising body mass index, current smoking, and presence of diabetes were each independent predictors of expander/implant loss. When controlling for all other identified confounders, use of acellular dermal matrix was associated with a significant increase in expander/implant loss (odds ratio, 1.42; 95 percent confidence interval, 1.04 to 1.94; p = 0.026). Thirty-day risk for expander/implant loss after tissue expander or implant-based breast reconstruction was 2.05 percent. Use of acellular dermal matrix was associated with a 0.7 percent absolute risk increase for expander/implant loss. Risk, III.

  4. Application of Bladder Acellular Matrix in Urinary Bladder Regeneration: The State of the Art and Future Directions

    Directory of Open Access Journals (Sweden)

    Marta Pokrywczynska

    2015-01-01

    Full Text Available Construction of the urinary bladder de novo using tissue engineering technologies is the “holy grail” of reconstructive urology. The search for the ideal biomaterial for urinary bladder reconstruction has been ongoing for decades. One of the most promising biomaterials for this purpose seems to be bladder acellular matrix (BAM. In this review we determine the most important factors, which may affect biological and physical properties of BAM and its regeneration potential in tissue engineered urinary bladder. We also point out the directions in modification of BAM, which include incorporation of exogenous growth factors into the BAM structure. Finally, we discuss the results of the urinary bladder regeneration with cell seeded BAM.

  5. Bioengineering Human Myocardium on Native Extracellular Matrix

    Science.gov (United States)

    Guyette, Jacques P.; Charest, Jonathan M; Mills, Robert W; Jank, Bernhard J.; Moser, Philipp T.; Gilpin, Sarah E.; Gershlak, Joshua R.; Okamoto, Tatsuya; Gonzalez, Gabriel; Milan, David J.; Gaudette, Glenn R.; Ott, Harald C.

    2015-01-01

    Rationale More than 25 million individuals suffer from heart failure worldwide, with nearly 4,000 patients currently awaiting heart transplantation in the United States. Donor organ shortage and allograft rejection remain major limitations with only about 2,500 hearts transplanted each year. As a theoretical alternative to allotransplantation, patient-derived bioartificial myocardium could provide functional support and ultimately impact the treatment of heart failure. Objective The objective of this study is to translate previous work to human scale and clinically relevant cells, for the bioengineering of functional myocardial tissue based on the combination of human cardiac matrix and human iPS-derived cardiac myocytes. Methods and Results To provide a clinically relevant tissue scaffold, we translated perfusion-decellularization to human scale and obtained biocompatible human acellular cardiac scaffolds with preserved extracellular matrix composition, architecture, and perfusable coronary vasculature. We then repopulated this native human cardiac matrix with cardiac myocytes derived from non-transgenic human induced pluripotent stem cells (iPSCs) and generated tissues of increasing three-dimensional complexity. We maintained such cardiac tissue constructs in culture for 120 days to demonstrate definitive sarcomeric structure, cell and matrix deformation, contractile force, and electrical conduction. To show that functional myocardial tissue of human scale can be built on this platform, we then partially recellularized human whole heart scaffolds with human iPSC-derived cardiac myocytes. Under biomimetic culture, the seeded constructs developed force-generating human myocardial tissue, showed electrical conductivity, left ventricular pressure development, and metabolic function. Conclusions Native cardiac extracellular matrix scaffolds maintain matrix components and structure to support the seeding and engraftment of human iPS-derived cardiac myocytes, and enable

  6. 人脂肪来源干细胞与膀胱脱细胞基质-丝素蛋白双层支架的生物相容性研究%Human Adipose-De rived Stem Cells and its Biocompatibility with Bladder Acellular Matrix Graft-Silk Fibroin Bilayer Scaffold

    Institute of Scientific and Technical Information of China (English)

    赵阳; 吴稼晟; 周哲; 周娟; 张明; 李伟; 王忠; 孙康; 卢慕峻

    2014-01-01

    Objective To observe the growth of human adipose-derived stem cells (hASCs) in bladder acellular matrix graft-silk fibroin (BAMG-SF) bilayer scaffold and to analyze the biological compatibility of BAMG-SF with hASCs. Methods hASCs were isolated from human subcutaneous adipose tissue after collagenase digesting, filtrating and centrifuging, then cultured in the leaching solution of BAMG-SF. The cytotoxicity of scaffold was evaluated by CCK-8 cell viability assay, and the growth curves were also observed. Surface morphology on BAMG-SF was observed by scanning electron microscopy (SEM). The hASCs of passage 3 were seeded onto the BAMG-SF bilayer scaffolds for 1 week, then the BAMG-SF bilayer scaffolds seeded with hASCs were transplanted into nude mouse for 1 week or 2 weeks. The growth of cells in BAMG-SF biomaterials was observed by HE staining. The species origin of these cells in the BAMG-SF scaffolds cultured in vivo was detected by Immunofluorescence. Results hASCs maintained high proliferation rate in the leaching solution of BAMG-SF and the BAMG-SF scaffolds were nontoxic absolutely. According to the growth curves of hASCs cultured in the leaching solution of the BAMG-SF and DMEM, BAMG-SF scaffolds were conducive to the growth of hASCs. The histological study found that hASCs could grow into the space of the BAMG-SF scaffolds after cultured in vitro and in vivo. There were more cells in the scaffolds cultured in vivo than in vitro. Immuno-fluorescence suggested that some of the cells inside the scaffolds were hASCs. Conclusion BAMG-SF bilayer scaffolds are nontoxic and have a good biocompatibility with hASCs, which can be used as a vehicle for hASCs in bladder defect reconstruction.%目的:观察人脂肪来源干细胞(Human adipose derived stem cells,hASCs)在膀胱黏膜下脱细胞基质-丝素蛋白(Bladder acellular matrix graft-silk fibroin,BAMG-SF)双层支架材料中的生长情况,分析其生物相容性。方法取hASCs,置

  7. Construction and evaluation of acellular matrix for ureter tissue engineering%输尿管无细胞基质移植物的制备和评价

    Institute of Scientific and Technical Information of China (English)

    沈海波; 潘俊; 陈方

    2009-01-01

    的种子细胞具备一定的生长能力.%BACKGROUND:Compared to small-intestine submucosa,acellular vascular grafts have natural tubic structure,which is similar to ureter.When it is used as replacement for ureter,end-to-end anastomosis is used.It is characterized by simple operation,smooth vessel wall,collection and preparation method.OBJECTIVE:To prepare acellular vascular matrix as scaffold in tissue-engineered ureter in vitro.DESIGN,TIME AND SETTING:The observational experiment was performed at the Animal Experimental Center of Xinhua Hospital Affiliated to School of Medicine,Shanghai Jiao Tong University from September 2006 to June 2008.MATERIALS:Swines were supplied by Shanghai Song Lian experimental animals company.Eight healthy adult rats were supplied by Animal Experimental Center Affiliated to School of Medicine,Shanghai Jiao Tong University for the animal toxicity study of acellular vascular matrix.METHODS:Swine carotid artery membrana was removed and placed in phosphate-buffed saline(PBS,pH 7.1).The tissue was stirred at 4 ℃ with 0.5%sodium dodecylsulfate for 24 hours.Then the tissue was treated by double distilled water for 1 week at 4 ℃.Double distilled water was changed twice every day.Vessel with muscle were digested in mixed digestive juice at 37 ℃ for 2 hours before washing.The acellular matrix was stored in penicillin and streptomycin solution at 4 ℃.MAIN OUTCOME MEASURES:The components of acellular scaffold was investigated by optical and electron microscopes.Allogenic endothelial progenitor cells following proliferation were transplanted into acellular vascular matrix to observe cell growth.Animal toxicity study of acellular vascular matrix was performed.Tensile force study was employed to understand the contractility of acellular vascular matrix.RESULTS:Acellular vascular matrix was without cell component.Acellular vascular matrix was mainly composed of collagen.Under scanning electron microscope,cells and cell debris were not found

  8. Dual plane penile augmentation with human acellular dermal matrix through penile lengthening incision%阴茎延长同期行脱细胞异体真皮基质补片双平面植入阴茎增粗术

    Institute of Scientific and Technical Information of China (English)

    吴小蔚; 简麒超; 董玉林; 龙道畴

    2011-01-01

    目的 探讨阴茎延长同期行脱细胞异体真皮基质(acellular dermal matrix,ADM)补片双平面植入阴茎增粗术的方法与效果.方法 采用阴茎根部倒V形切口,离断阴茎浅悬韧带后,沿阴茎纵轴切开Dartos筋膜,在其深面向远端分离.距冠状沟1.5~2.0 cm处环形切开Buck筋膜,将补片前部植于Buck筋膜与白膜间,后部植于Dartos筋膜与Buck筋膜间.缝合Dartos筋膜切口,V-Y成形术闭合阴茎根部切口.结果 35例术后无1例发生阴茎皮肤坏死、补片外露并发症.25例获随访6~24个月,对阴茎外形均感满意;无1例出现补片移位或皱褶、阴茎头感觉异常;其中21例已婚者均感性生活满意.结论 经阴茎根部切口行脱细胞异体真皮基质补片双平面植入阴茎增粗术,通过调整补片植入层次,在确保补片足够的组织覆盖及阴茎皮肤血供情况下,在Ⅰ期内行延长并增粗阴茎术,具有并发症少、疗效满意的优点.%Objective To illustrate the details and effects of a new technique of penile augmentation-a dual plane approach to enhance the penile girth with human acellular dermal matrix (ADM)through the incision on the dorsal penile root.Methods Firstly,a reversed V incision was made at the dorsal root of the penis and the superficial suspensory ligament of the penis was released.A Dartos fascia incision was then made and the plan between Dartos fascia and Buck's fascia was dorsally dissected toward the coronary sulcus.A Buck's fascia incision was made 1.5-2 cm from the coronary sulcus and the fascia was undermined distally.One or two sheets of ADM was dorsally placed by a dual plane method which combined partial sub-Buck's fascia plane and partial sub-Dartos fascia plane to enhance the penile circumference.Finally,the Dartos fascia incision was closed and followed by the closure of the wound with V-Y advancement.Results A total of 35 patients underwent dual plane penile augmentation No dorsal penile skin necrosis

  9. Does Acellular Dermal Matrix Thickness Affect Complication Rate in Tissue Expander Based Breast Reconstruction?

    Directory of Open Access Journals (Sweden)

    Jessica F. Rose

    2016-01-01

    Full Text Available Background. While the benefits of using acellular dermal matrices (ADMs in breast reconstruction are well described, their use has been associated with additional complications. The purpose of this study was to determine if ADM thickness affects complications in breast reconstruction. Methods. A retrospective chart review was performed including all tissue expander based breast reconstructions with AlloDerm (LifeCell, Branchburg, NJ over 4 years. We evaluated preoperative characteristics and assessed postoperative complications including seroma, hematoma, infection, skin necrosis, and need for reintervention. We reviewed ADM thickness and time to Jackson-Pratt (JP drain removal. Results. Fifty-five patients underwent 77 ADM-associated tissue expander based breast reconstructions, with average age of 48.1 years and average BMI of 25.9. Average ADM thickness was 1.21 mm. We found higher complication rates in the thick ADM group. Significant associations were found between smokers and skin necrosis (p<0.0001 and seroma and prolonged JP drainage (p=0.0004; radiated reconstructed breasts were more likely to suffer infections (p=0.0085, and elevated BMI is a significant predictor for increased infection rate (p=0.0037. Conclusion. We found a trend toward increased complication rates with thicker ADMs. In the future, larger prospective studies evaluating thickness may provide more information.

  10. Acellular Dermal Matrix in Reconstructive Breast Surgery: Survey of Current Practice among Plastic Surgeons

    Science.gov (United States)

    Ibrahim, Ahmed M. S.; Koolen, Pieter G. L.; Ashraf, Azra A.; Kim, Kuylhee; Mureau, Marc A. M.; Lee, Bernard T.

    2015-01-01

    Background: Acellular dermal matrices (ADMs) in plastic surgery have become increasingly popular particularly for breast reconstruction. Despite their advantages, questions exist regarding their association with a possible increased incidence of complications. We describe a collective experience of plastic surgeons’ use of ADMs in reconstructive breast surgery using an internet-based survey. Methods: Members of the American Society of Plastic Surgeons were recruited through voluntary, anonymous participation in an online survey. The web-based survey garnered information about participant demographics and their experience with ADM use in breast reconstruction procedures. After responses were collected, all data were anonymously processed. Results: Data were ascertained through 365 physician responses of which 99% (n = 361) completed the survey. The majority of participants were men (84.5%) between 51 and 60 years (37.4%); 84.2% used ADM in breast reconstruction, including radiated patients (79.7%). ADM use was not favored for nipple reconstruction (81.5%); 94.6% of participants used drains, and 87.8% administered antibiotics postoperatively. The most common complications were seroma (70.9%) and infection (16%), although 57.4% claimed anecdotally that overall complication rate was unchanged after incorporating ADM into their practice. High cost was a deterrent for ADM use (37.5%). Conclusions: Plastic surgeons currently use ADM in breast reconstruction for both immediate and staged procedures. Of those responding, a majority of plastic surgeons will incorporate drains and use postoperative antibiotics for more than 48 hours. PMID:25973359

  11. An evaluation of resource utilisation of single stage porcine acellular dermal matrix assisted breast reconstruction: A comparative study.

    Science.gov (United States)

    Kilchenmann, Ashley J R; Lardi, Alessia M; Ho-Asjoe, Mark; Junge, Klaus; Farhadi, Jian

    2014-12-01

    To evaluate resource utilization of single stage porcine acellular dermal matrix (ADM) assisted breast reconstruction compared with tissue expander (TE), latissimus dorsi flap and implant (LD/I) and latissimus dorsi flap and TE (LD/TE) reconstructive techniques. Clinical data was collected for length of stay, operative time, additional hospitalisations and operative procedures, and outpatient appointments for 101 patients undergoing unilateral implant based breast reconstruction. Resources utilised by ADM (Strattice Reconstructive Tissue Matrix™) patients were analysed and compared to the resource usage of traditional techniques. 25 patients undergoing single stage ADM (ADM/I) were compared with 27 having TE, 32 having LD/I and 17 having LD/TE reconstructions. Follow up was 24 months. Compared to TE, ADM/I had similar length of stay and operative time, lower rate and number of additional procedures, fewer, shorter re-admissions (p reconstructions in both complication-free and complicated settings over a 24-month period, despite requiring aesthetic revision in 60.9% of patients. Compared to LD/I, resource utilisation was commensurate in complication-free and complicated settings. Copyright © 2014 Elsevier Ltd. All rights reserved.

  12. Comparison of Achilles tendon repair techniques in a sheep model using a cross-linked acellular porcine dermal patch and platelet-rich plasma fibrin matrix for augmentation.

    Science.gov (United States)

    Sarrafian, Tiffany L; Wang, Hali; Hackett, Eileen S; Yao, Jian Q; Shih, Mei-Shu; Ramsay, Heather L; Turner, A Simon

    2010-01-01

    The primary goal of this study was to evaluate a cross-linked acellular porcine dermal patch (APD), as well as platelet-rich plasma fibrin matrix (PRPFM), for repair of acute Achilles tendon rupture in a sheep model. The 2 surgically transected tendon ends were reapproximated in groups 1 and 2, whereas a gap was left between the tendon ends in group 3. APD was used to reinforce the repair in group 2, and autologous PRPFM was used to fill the gap, which was also reinforced with APD, in group 3. All sheep were humanely euthanized at 24 weeks after the repair, and biomechanical and histological testing were performed. Tensile strength testing showed a statistically significant difference in elongation between the operated limb and the unoperated contralateral limb in groups 1 and 3, but not in group 2. All operated tendons appeared healed with no apparent fibrosis under light and polarized microscopy. In group 1, all surgical separation sites were identifiable, and healing occurred via increasing tendon thickness. In group 2, healing occurred with new tendon fibers across the separation, without increasing tendon thickness in 2 out of 6 animals. Group 3 showed complete bridging of the gap, with no change in tendon thickness in 2 out of 6 animals. In groups 2 and 3, peripheral integration of the APD to tendon fibers was observed. These findings support the use of APD, alone or with PRPFM, to augment Achilles tendon repair in a sheep model.

  13. Cellular versus acellular matrix devices in treatment of diabetic foot ulcers: study protocol for a comparative efficacy randomized controlled trial

    Directory of Open Access Journals (Sweden)

    Lev-Tov Hadar

    2013-01-01

    Full Text Available Abstract Background Diabetic foot ulcers (DFUs represent a significant source of morbidity and an enormous financial burden. Standard care for DFUs involves systemic glucose control, ensuring adequate perfusion, debridement of nonviable tissue, off-loading, control of infection, local wound care and patient education, all administered by a multidisciplinary team. Unfortunately, even with the best standard of care (SOC available, only 24% or 30% of DFUs will heal at weeks 12 or 20, respectively. The extracellular matrix (ECM in DFUs is abnormal and its impairment has been proposed as a key target for new therapeutic devices. These devices intend to replace the aberrant ECM by implanting a matrix, either devoid of cells or enhanced with fibroblasts, keratinocytes or both as well as various growth factors. These new bioengineered skin substitutes are proposed to encourage angiogenesis and in-growth of new tissue, and to utilize living cells to generate cytokines needed for wound repair. To date, the efficacy of bioengineered ECM containing live cellular elements for improving healing above that of a SOC control group has not been compared with the efficacy of an ECM devoid of cells relative to the same SOC. Our hypothesis is that there is no difference in the improved healing effected by either of these two product types relative to SOC. Methods/Design To test this hypothesis we propose a randomized, single-blind, clinical trial with three arms: SOC, SOC plus Dermagraft® (bioengineered ECM containing living fibroblasts and SOC plus Oasis® (ECM devoid of living cells in patients with nonhealing DFUs. The primary outcome is the percentage of subjects that achieved complete wound closure by week 12. Discussion If our hypothesis is correct, then immense cost savings could be realized by using the orders-of-magnitude less expensive acellular ECM device without compromising patient health outcomes. The article describes the protocol proposed to test

  14. Decreased Laminin Expression by Human Lung Epithelial Cells and Fibroblasts Cultured in Acellular Lung Scaffolds from Aged Mice.

    Directory of Open Access Journals (Sweden)

    Lindsay M Godin

    Full Text Available The lung changes functionally and structurally with aging. However, age-related effects on the extracellular matrix (ECM and corresponding effects on lung cell behavior are not well understood. We hypothesized that ECM from aged animals would induce aging-related phenotypic changes in healthy inoculated cells. Decellularized whole organ scaffolds provide a powerful model for examining how ECM cues affect cell phenotype. The effects of age on ECM composition in both native and decellularized mouse lungs were assessed as was the effect of young vs old acellular ECM on human bronchial epithelial cells (hBECs and lung fibroblasts (hLFs. Native aged (1 year lungs demonstrated decreased expression of laminins α3 and α4, elastin and fibronectin, and elevated collagen, compared to young (3 week lungs. Proteomic analyses of decellularized ECM demonstrated similar findings, and decellularized aged lung ECM contained less diversity in structural proteins compared to young ECM. When seeded in old ECM, hBECs and hLFs demonstrated lower gene expression of laminins α3 and α4, respectively, as compared to young ECM, paralleling the laminin deficiency of aged ECM. ECM changes appear to be important factors in potentiating aging-related phenotypes and may provide clues to mechanisms that allow for aging-related lung diseases.

  15. Decreased Laminin Expression by Human Lung Epithelial Cells and Fibroblasts Cultured in Acellular Lung Scaffolds from Aged Mice.

    Science.gov (United States)

    Godin, Lindsay M; Sandri, Brian J; Wagner, Darcy E; Meyer, Carolyn M; Price, Andrew P; Akinnola, Ifeolu; Weiss, Daniel J; Panoskaltsis-Mortari, Angela

    2016-01-01

    The lung changes functionally and structurally with aging. However, age-related effects on the extracellular matrix (ECM) and corresponding effects on lung cell behavior are not well understood. We hypothesized that ECM from aged animals would induce aging-related phenotypic changes in healthy inoculated cells. Decellularized whole organ scaffolds provide a powerful model for examining how ECM cues affect cell phenotype. The effects of age on ECM composition in both native and decellularized mouse lungs were assessed as was the effect of young vs old acellular ECM on human bronchial epithelial cells (hBECs) and lung fibroblasts (hLFs). Native aged (1 year) lungs demonstrated decreased expression of laminins α3 and α4, elastin and fibronectin, and elevated collagen, compared to young (3 week) lungs. Proteomic analyses of decellularized ECM demonstrated similar findings, and decellularized aged lung ECM contained less diversity in structural proteins compared to young ECM. When seeded in old ECM, hBECs and hLFs demonstrated lower gene expression of laminins α3 and α4, respectively, as compared to young ECM, paralleling the laminin deficiency of aged ECM. ECM changes appear to be important factors in potentiating aging-related phenotypes and may provide clues to mechanisms that allow for aging-related lung diseases.

  16. Managing real world venous leg ulcers with fetal bovine acellular dermal matrix: a single centre retrospective study.

    Science.gov (United States)

    Paredes, J A; Bhagwandin, S; T, Polanco; Lantis, J C

    2017-10-01

    As compression treatment offers moderate improvement, especially to recurrent venous leg ulcers (VLUs), several alternative therapies using cellular based and/or tissue-derived products (CTPs) have emerged from bovine, porcine, and equine sources. Our aim was to look at the effect of a CTP in 'real-life' VLUs. This study looked at complex patients with chronic, large wounds in a single-centre retrospective review. All patients were treated with fetal bovine acellular dermal matrix (FBADM) for VLUs at our outpatient urban wound care programme. A total of 40 wounds in 33 patients were analysed. At week four, 6% of wounds were closed; at week eight, 9% were closed; at week 12, 25% were closed; and at week 16, 38% of wounds were closed. The median time to wound closure was 67 days (range: 23-100 days) and the median percent wound closure through re-epithelialisation was 11% per week (range: 7-30% per week). At 4 weeks the median area reduction of all wounds was 23.5%, with 40% of VLUs having a ≥40% area reduction at the same point in time. There are limitations to any retrospective review; however; patients deemed to have a limited chance of closure at 4 months did better than expected, either healing or having a wound area reduction at 16 weeks, making their wound care much easier. Prospective studies should be conducted to optimise the treatment algorithm to determine if better clinical outcomes can be obtained for the 'real-life' VLU population.

  17. Tissue-engineered conduit using bladder acellular matrix and bladder epithelial cells for urinary diversion in rabbits

    Institute of Scientific and Technical Information of China (English)

    LIAO Wen-biao; SONG Chao; LI Yong-wei; YANG Si-xing; MENG Lin-chao; LI Xin-hui

    2013-01-01

    Background For muscle invasive bladder cancer,radical cystectomy is the most effective treatment now and urinary diversion is often necessary.The use of intestinal tissue for urinary diversion is frequently associated with complications.In this study,we aimed to make a tissue-engineered conduit (TEC) using bladder epithelial cells and bladder acellular matrix (BAM) for urinary diversion in rabbits.Methods Bladder epithelial cells of rabbit were cultivated and expanded in vitro,then seeded on BAM,and cultured for 7 days.Then cell-seeded graft was used to make TEC.In the experimental group,most of bladder of the rabbit was removed while bladder trigone was retained.The proximal end of TEC was anastomosed with bladder trigone and the distal end was anastomosed with the abdominal stoma.In the control group,TEC was made using unseeded BAM.Haematoxylin and eosin staining was conducted,respectively,at 1,2,4,and 8 weeks postoperatively.Immunohistochemistry was performed 8 weeks postoperatively.Intravenous urography,retrograde pyelography,and cystoscopy of TEC were made at 12 weeks postoperatively.Results All animals were alive in the experimental group.Haematoxylin and eosin staining showed epithelial coverage in TEC.Immunohistochemistry showed anti-cytokeratin AE1/AE3 antibody and anti-ZO1 antibody positive,confirming there were mature and functional epithelial cells on the lumen of TEC.Retrograde pyelography and intravenous urography showed that TEC developed well and that there was no obstruction.In the control group,four rabbits were dead within 2 weeks and scar formation,atresia,and severe hydronephrosis were found.Conclusions We successfully made TEC using BAM and bladder epithelial cells for urinary diversion in rabbits.The lumen of this new TEC covered mature epithelial cells and could prevent urinary extravasation.

  18. Surgisis acellular collagen matrix in aesthetic and reconstructive plastic surgery soft tissue applications.

    Science.gov (United States)

    Centeno, Robert F

    2009-04-01

    Tissue engineering in aesthetic and reconstructive plastic surgery remains an elusive goal. The advent of Surgisis extracellular collagen matrix and its performance characteristics suggest that the use of a bioengineered tissue substitute can meet some of our reconstructive requirements. Incorporation and replacement by host tissue with minimal allergic or immune response seems to be achievable today. The ability to engineer the device, the ready availability of substrate, and its cost effectiveness support the use of Surgisis in aesthetic and reconstructive plastic surgery applications. Future product innovations and engineering seem promising. The permanent role of Surgisis in aesthetic and reconstructive plastic surgery will be determined by its documented long-term performance.

  19. Healing rates for challenging rotator cuff tears utilizing an acellular human dermal reinforcement graft

    Science.gov (United States)

    Agrawal, Vivek

    2012-01-01

    Purpose: This study presents a retrospective case series of the clinical and structural outcomes (1.5 T MRI) of arthroscopic rotator cuff repair with acellular human dermal graft reinforcement performed by a single surgeon in patients with large, massive, and previously repaired rotator cuff tears. Materials and Methods: Fourteen patients with mean anterior to posterior tear size 3.87 ± 0.99 cm (median 4 cm, range 2.5–6 cm) were enrolled in the study and were evaluated for structural integrity using a high-field (1.5 T) MRI at an average of 16.8 months after surgery. The Constant-Murley scores, the Flexilevel Scale of Shoulder Function (Flex SF), scapular plane abduction, and strength were analyzed. Results: MRI results showed that the rotator cuff repair was intact in 85.7% (12/14) of the patients studied. Two patients had a Sugaya Type IV recurrent tear (2 of 14; 14.3%), which were both less than 1 cm. The Constant score increased from a preoperative mean of 49.72 (range 13–74) to a postoperative mean of 81.07 (range 45–92) (P value = 0.009). Flexilevel Scale of Shoulder Function (Flex SF) Score normalized to a 100-point scale improved from a preoperative mean of 53.69 to a postoperative mean of 79.71 (P value = 0.003). The Pain Score improved from a preoperative mean of 7.73 to a postoperative mean of 13.57 (P value = 0.008). Scapular plane abduction improved from a preoperative mean of 113.64° to a postoperative mean of 166.43° (P value = 0.010). The strength subset score improved from a preoperative mean of 1.73 kg to a postoperative mean of 7.52 kg (P value = 0.006). Conclusions: This study presents a safe and effective technique that may help improve the healing rates of large, massive, and revision rotator cuff tears with the use of an acellular human dermal allograft. This technique demonstrated favorable structural healing rates and statistically improved functional outcomes in the near term. Level of Evidence: 4. Retrospective case series. PMID

  20. Three-dimensional Reconstruction of the Microstructure of Human Acellular Nerve Allograft

    Science.gov (United States)

    Zhu, Shuang; Zhu, Qingtang; Liu, Xiaolin; Yang, Weihong; Jian, Yutao; Zhou, Xiang; He, Bo; Gu, Liqiang; Yan, Liwei; Lin, Tao; Xiang, Jianping; Qi, Jian

    2016-01-01

    The exact inner 3D microstructure of the human peripheral nerve has been a mystery for decades. Therefore, it has been difficult to solve several problems regarding peripheral nerve injury and repair. We used high-resolution X-ray computed microtomography (microCT) to scan a freeze-dried human acellular nerve allograft (hANA). The microCT images were then used to reconstruct a 3D digital model, which was used to print a 3D resin model of the nerve graft. The 3D digital model of the hANA allowed visualization of all planes. The magnified 3D resin model clearly showed the nerve bundles and basement membrane tubes of the hANA. Scanning electron microscopy (SEM) was used to analyse the microstructure of the hANA. Compared to the SEM images, the microCT image clearly demonstrated the microstructure of the hANA cross section at a resolution of up to 1.2 μm. The 3D digital model of the hANA facilitates a clear and easy understanding of peripheral nerve microstructure. Furthermore, the enlarged 3D resin model duplicates the unique inner structure of each individual hANA. This is a crucial step towards achieving 3D printing of a hANA or nerve that can be used as a nerve graft. PMID:27476584

  1. Time-dependent bladder tissue regeneration using bilayer bladder acellular matrix graft-silk fibroin scaffolds in a rat bladder augmentation model.

    Science.gov (United States)

    Zhao, Yang; He, Yi; Zhou, Zhe; Guo, Jian-hua; Wu, Jia-sheng; Zhang, Ming; Li, Wei; Zhou, Juan; Xiao, Dong-dong; Wang, Zhong; Sun, Kang; Zhu, Ying-jian; Lu, Mu-jun

    2015-09-01

    With advances in tissue engineering, various synthetic and natural biomaterials have been widely used in tissue regeneration of the urinary bladder in rat models. However, reconstructive procedures remain insufficient due to the lack of appropriate scaffolding, which should provide a waterproof barrier function and support the needs of various cell types. To address these problems, we have developed a bilayer scaffold comprising a porous network (silk fibroin [SF]) and an underlying natural acellular matrix (bladder acellular matrix graft [BAMG]) and evaluated its feasibility and potential for bladder regeneration in a rat bladder augmentation model. Histological (hematoxylin and eosin and Masson's trichrome staining) and immunohistochemical analyses demonstrated that the bilayer BAMG-SF scaffold promoted smooth muscle, blood vessel, and nerve regeneration in a time-dependent manner. At 12weeks after implantation, bladders reconstructed with the BAMG-SF matrix displayed superior structural and functional properties without significant local tissue responses or systemic toxicity. These results demonstrated that the bilayer BAMG-SF scaffold may be a promising scaffold with good biocompatibility for bladder regeneration in the rat bladder augmentation model.

  2. Healing rate and autoimmune safety of full-thickness wounds treated with fish skin acellular dermal matrix versus porcine small-intestine submucosa: a noninferiority study.

    Science.gov (United States)

    Baldursson, Baldur Tumi; Kjartansson, Hilmar; Konrádsdóttir, Fífa; Gudnason, Palmar; Sigurjonsson, Gudmundur F; Lund, Sigrún Helga

    2015-03-01

    A novel product, the fish skin acellular dermal matrix (ADM) has recently been introduced into the family of biological materials for the treatment of wounds. Hitherto, these products have been produced from the organs of livestock. A noninferiority test was used to compare the effect of fish skin ADM against porcine small-intestine submucosa extracellular matrix in the healing of 162 full-thickness 4-mm wounds on the forearm of 81 volunteers. The fish skin product was noninferior at the primary end point, healing at 28 days. Furthermore, the wounds treated with fish skin acellular matrix healed significantly faster. These results might give the fish skin ADM an advantage because of its environmental neutrality when compared with livestock-derived products. The study results on these acute full-thickness wounds might apply for diabetic foot ulcers and other chronic full-thickness wounds, and the shorter healing time for the fish skin-treated group could influence treatment decisions. To test the autoimmune reactivity of the fish skin, the participants were tested with the following ELISA (enzyme-linked immunosorbent assay) tests: RF, ANA, ENA, anti ds-DNA, ANCA, anti-CCP, and anticollagen I and II. These showed no reactivity. The results demonstrate the claims of safety and efficacy of fish skin ADM for wound care.

  3. A Meta-analysis of Postoperative Complications of Tissue Expander/Implant Breast Reconstruction Using Acellular Dermal Matrix.

    Science.gov (United States)

    Zhao, Xiangyi; Wu, Xiaowei; Dong, Jie; Liu, Yingying; Zheng, Liang; Zhang, Liming

    2015-12-01

    Acellular dermal matrix (ADM) is commonly used for tissue expander/implant breast (TE/I-based) reconstruction. But the relation between ADM and postoperative complications remains controversial. A few meta-analyses were conducted in 2011-2012 and the result revealed that ADM can increase the risk of complications. The purpose of our study is to offer updated evidence for ADM clinical application by analyzing the effect of ADM on complications of TE/I-based breast reconstruction. The literature published from January 2010 to February 2015 was searched in EMbase, Medline, Science Direct, the Cochrane Central Register of Controlled Trials (CENTRAL), CBMdisc, CNKI, VIP, and the references of those included studies were also searched by hand. According to inclusive criteria, 11 studies were selected and the values were extracted from the included literature. Complications with four different categories assigned for overall complications, infection, hematoma/seroma, and explantation were collected. RevMan 5.1 was used for meta-analysis. The evidence level was assessed by using the GRADE system. Eleven published studies were included. The results showed that compared to the control group, the ADM group increased the rate of overall complications (OR = 1.33, 95% CI 1.03-1.70, p = 0.03), infection (OR = 1.47, 95% CI 1.04-2.06, p = 0.03), hematoma/seroma (OR = 1.66, 95% CI 1.13-2.44, p = 0.01), but there was no significant difference in explantation (OR = 1.37, 95% CI 0.89-2.11, p = 0.15). Based on the GRADE system, all the evidence was at level C and weak recommendation. In TE/I-based breast reconstruction, ADM increased the incidence of overall complications, infection, and hematoma/seroma; the incidence of explantation remains unknown. For the poor quality of the original studies, a prudent choice is suggested; and more high-quality, large-sample studies are needed. This journal requires that authors assign a level of evidence to each submission to which Evidence

  4. The biomechanical behavior and host response to porcine-derived small intestine submucosa, pericardium and dermal matrix acellular grafts in a rat abdominal defect model.

    Science.gov (United States)

    Zhang, Jian; Wang, Guan Yu; Xiao, Yi Pin; Fan, Lie Ying; Wang, Qiang

    2011-10-01

    Several porcine-derived acellular biologic grafts are increasingly used in abdominal wall reconstruction due to the limitations of synthetic meshes in many clinical situations. However, relatively little is known so far about their comparative mechanical characteristics and performance after defect repair. We therefore investigated three most commonly used porcine-derived acellular biomaterials, small intestine submucosa (P-SIS), pericardium (P-PC) and acellular dermal matrix (P-ADM) immediately after prepared, and their effectiveness, biomechanical and histological characteristics in repairing full-thickness abdominal defect in a rat model. P-PC had the best native performance in the burst strength, tensile strength and ball burst among the three porcine-derived scaffolds. P-SIS showed a significantly higher water vapor transmission in comparison with P-PC or P-ADM. Abdominal wall defects in rats were all satisfied repaired with P-SIS, P-PC or P-ADM. No laxity or fistula was observed in the repaired abdominal wall in the P-SIS group up to 8 weeks after surgery. However, there was a tendency for high postoperative abdominal eventration in the P-ADM and P-PC groups as compared with the P-SIS group. With regard to overall aspects of the postoperative laxity, intra-abdominal adhesion formation, tensile stress, stretchability, and degree of tissue ingrowth in terms of collagen deposition and neovascularization, P-SIS exhibits clear advantages over P-PC as well as P-ADM after large abdominal wall defect reconstruction.

  5. Combined periodontal and restorative approach to the treatment of gingival recessions with noncarious cervical lesions: a case treated with acellular dermal matrix allograft and compomer restorations.

    Science.gov (United States)

    Efeoğlu, Ahmet; Hanzade, Mete; Sari, Esra; Alpay, Hande; Karakaş, Ozan; Koray, Fatma

    2012-08-01

    Treatment of gingival recessions has become one of the most challenging procedures in periodontal plastic surgery. Various surgical options with predictable outcomes are available, but in cases with cervical lesions or restorations, optimal functional and esthetic results may require the combination of periodontal and restorative procedures. In this case report, one patient treated with acellular dermal matrix allograft and a coronally positioned flap in combination with compomer cervical restorations is presented. Clinical parameters were recorded immediately prior to surgery and after 12 months. Postoperatively, significant root coverage, reductions in probing depths, and gains in clinical attachment were observed. The final clinical results, esthetics, color match, and tissue contours were acceptable to both the patient and clinicians.

  6. A Microbiological and Ultrastructural Comparison of Aseptic versus Sterile Acellular Dermal Matrix as a Reconstructive Material and a Scaffold for Stem Cell Ingrowth.

    Science.gov (United States)

    Mendenhall, Shaun D; Schmucker, Ryan W; Daugherty, Timothy H F; Kottwitz, Katherine M; Reichensperger, Joel D; Koirala, Janak; Cederna, Paul S; Neumeister, Michael W

    2017-07-01

    Recent data suggest an increased risk for infection when acellular dermal matrix is used in breast reconstruction. This may be because some acellular dermal matrices are actually not terminally sterilized but are instead "aseptically processed." This study evaluates aseptic and sterile matrices for evidence of bacterial contamination and whether or not terminal sterilization affects matrix collagen architecture and stem cell ingrowth. Five separate samples of 14 different matrices were analyzed by fluorescent in situ hybridization using a bacterial DNA probe to detect bacterial DNA on the matrices. Separate samples were incubated for bacteria, acid-fast bacilli, and fungi for 2 to 6 weeks to detect living organisms. The impact of terminal sterilization on the collagen network and stem cell ingrowth on the matrices was then assessed. Traces of bacterial DNA were encountered on all matrices, with more bacteria in the aseptic group compared with the sterile group (3.4 versus 1.6; p = 0.003). The number of positive cultures was the same between groups (3.8 percent). Electron microscopy demonstrated decreased collagen organization in the sterile group. Stem cell seeding on the matrices displayed a wide variation of cellular ingrowth between matrices, with no difference between aseptic and sterile groups (p = 0.2). Although there was more evidence of prior bacterial contamination on aseptically processed matrices compared with sterile matrices; clinical cultures did not differ between groups. Terminal sterilization does not appear to affect stem cell ingrowth but may come at the cost of damaging the collagen network. Therapeutic, V.

  7. Evaluation of the Antimicrobial Efficacy of a Novel Rifampin/Minocycline-Coated, Noncrosslinked Porcine Acellular Dermal Matrix Compared With Uncoated Scaffolds for Soft Tissue Repair.

    Science.gov (United States)

    Majumder, Arnab; Scott, Jeffrey R; Novitsky, Yuri W

    2016-10-01

    Background Despite meticulous aseptic technique and systemic antibiotics, bacterial colonization of mesh remains a critical issue in hernia repair. A novel minocycline/rifampin tyrosine-coated, noncrosslinked porcine acellular dermal matrix (XenMatrix AB) was developed to protect the device from microbial colonization for up to 7 days. The objective of this study was to evaluate the in vitro and in vivo antimicrobial efficacy of this device against clinically isolated methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli. Methods XenMatrix AB was compared with 5 existing uncoated soft tissue repair devices using in vitro methods of zone of inhibition (ZOI) and scanning electron microscopy (SEM) at 24 hours following inoculation with MRSA or E coli These devices were also evaluated at 7 days following dorsal implantation and inoculation with MRSA or E coli (60 male New Zealand white rabbits, n = 10 per group) for viable colony-forming units (CFU), abscess formation and histopathologic response, respectively. Results In vitro studies demonstrated a median ZOI of 36 mm for MRSA and 16 mm for E coli for XenMatrix AB, while all uncoated devices showed no inhibition of bacterial growth (0 mm). SEM also demonstrated no visual evidence of MRSA or E coli colonization on the surface of XenMatrix AB compared with colonization of all other uncoated devices. In vivo XenMatrix AB demonstrated complete inhibition of bacterial colonization, no abscess formation, and a reduced inflammatory response compared with uncoated devices. Conclusion We demonstrated that XenMatrix AB possesses potent in vitro and in vivo antimicrobial efficacy against clinically isolated MRSA and E coli compared with uncoated devices.

  8. Acellular dermal matrix and negative pressure wound therapy: a tissue-engineered alternative to free tissue transfer in the compromised host.

    Science.gov (United States)

    Menn, Zachary K; Lee, Edward; Klebuc, Michael J

    2012-02-01

    Free tissue transfer has revolutionized lower extremity reconstruction; however, its use in elderly patients with multiple medical problems can be associated with elevated rate s of perioperative morbidity and mortality. This study evaluates the use of acellular dermal matrix (ADM) in conjunction with negative pressure wound therapy (NPWT) and delayed skin graft application as an alternative to free tissue transfer in this compromised population. Bilayer, ADM (Integra, Plainsboro, NJ) was used in conjunction with NPWT (Wound V.A.C, Kinetic Concepts Inc., San Antonio, TX) to achieve vascularized coverage of complex lower extremity wounds with denuded tendon and bone in elderly, medically compromised patients. Following incorporation, the matrix was covered with split-thickness skin graft. Four patients (age range, 50 to 76 years) with multiple medical comorbidities were treated with the above protocol. The average time to complete vascularization of the matrix was 29 days. Definitive closure with split-thickness skin graft was achieved in three patients and one wound healed by secondary intention. No medical or surgical complications were encountered and stable soft tissue coverage was achieved in all patients. This early experience suggests that dermal substitute and NPWT with delayed skin graft application can provide a reasonable tissue-engineered alternative to free tissue transfer in the medically compromised individual.

  9. Preservation of micro-architecture and angiogenic potential in a pulmonary acellular matrix obtained using intermittent intra-tracheal flow of detergent enzymatic treatment

    Science.gov (United States)

    Maghsoudlou, Panagiotis; Georgiades, Fanourios; Tyraskis, Athanasios; Totonelli, Giorgia; Loukogeorgakis, Stavros P.; Orlando, Giuseppe; Shangaris, Panicos; Lange, Peggy; Delalande, Jean-Marie; Burns, Alan J.; Cenedese, Angelo; Sebire, Neil J.; Turmaine, Mark; Guest, Brogan N.; Alcorn, John F.; Atala, Anthony; Birchall, Martin A.; Elliott, Martin J.; Eaton, Simon; Pierro, Agostino; Gilbert, Thomas W.; De Coppi, Paolo

    2013-01-01

    Tissue engineering of autologous lung tissue aims to become a therapeutic alternative to transplantation. Efforts published so far in creating scaffolds have used harsh decellularization techniques that damage the extracellular matrix (ECM), deplete its components and take up to 5 weeks to perform. The aim of this study was to create a lung natural acellular scaffold using a method that will reduce the time of production and better preserve scaffold architecture and ECM components. Decellularization of rat lungs via the intratracheal route removed most of the nuclear material when compared to the other entry points. An intermittent inflation approach that mimics lung respiration yielded an acellular scaffold in a shorter time with an improved preservation of pulmonary micro-architecture. Electron microscopy demonstrated the maintenance of an intact alveolar network, with no evidence of collapse or tearing. Pulsatile dye injection via the vasculature indicated an intact capillary network in the scaffold. Morphometry analysis demonstrated a significant increase in alveolar fractional volume, with alveolar size analysis confirming that alveolar dimensions were maintained. Biomechanical testing of the scaffolds indicated an increase in resistance and elastance when compared to fresh lungs. Staining and quantification for ECM components showed a presence of collagen, elastin, GAG and laminin. The intratracheal intermittent decellularization methodology could be translated to sheep lungs, demonstrating a preservation of ECM components, alveolar and vascular architecture. Decellularization treatment and methodology preserves lung architecture and ECM whilst reducing the production time to 3 h. Cell seeding and in vivo experiments are necessary to proceed towards clinical translation. PMID:23727263

  10. Regenerative and Antibacterial Properties of Acellular Fish Skin Grafts and Human Amnion/Chorion Membrane: Implications for Tissue Preservation in Combat Casualty Care.

    Science.gov (United States)

    Magnusson, Skuli; Baldursson, Baldur Tumi; Kjartansson, Hilmar; Rolfsson, Ottar; Sigurjonsson, Gudmundur Fertram

    2017-03-01

    Improvised explosive devices and new directed energy weapons are changing warfare injuries from penetrating wounds to large surface area thermal and blast injuries. Acellular fish skin is used for tissue repair and during manufacturing subjected to gentle processing compared to biologic materials derived from mammals. This is due to the absence of viral and prion disease transmission risk, preserving natural structure and composition of the fish skin graft. The aim of this study was to assess properties of acellular fish skin relevant for severe battlefield injuries and to compare those properties with those of dehydrated human amnion/chorion membrane. We evaluated cell ingrowth capabilities of the biological materials with microscopy techniques. Bacterial barrier properties were tested with a 2-chamber model. The microstructure of the acellular fish skin is highly porous, whereas the microstructure of dehydrated human amnion/chorion membrane is mostly nonporous. The fish skin grafts show superior ability to support 3-dimensional ingrowth of cells compared to dehydrated human amnion/chorion membrane (p < 0.0001) and the fish skin is a bacterial barrier for 24 to 48 hours. The unique biomechanical properties of the acellular fish skin graft make it ideal to be used as a conformal cover for severe trauma and burn wounds in the battlefield. Reprint & Copyright © 2017 Association of Military Surgeons of the U.S.

  11. Acellular Dermal Matrix (Permacol®) for Heterologous Immediate Breast Reconstruction after Skin-Sparing Mastectomy in Patients with Breast Cancer: A Single-Institution Experience and a Review of the Literature

    OpenAIRE

    Knabben, Laura; Kanagalingam, Gowthami; Imboden, Sara; Günthert, Andreas R.

    2017-01-01

    Objective Skin-sparing mastectomy (SSM) with immediate heterologous reconstruction is a safe oncological option in surgical therapy of early breast cancer. Permacol® is an acellular dermal matrix (ADM) placed between the implant and the skin to improve lower pole projection and implant coverage. The aim of our study was to evaluate the outcome with a focus on patient satisfaction after 6 months and to analyze physical changes of ADM. Methods 10 patients who underwent SSM with Perma...

  12. Application of acellular dermal matrix embedded in socket after wisdom tooth extraction%脱细胞异体真皮组织补片在智齿拔除中的应用

    Institute of Scientific and Technical Information of China (English)

    白忠诚; 施生根; 李莉莉; 牛忠英; 张艳茹

    2011-01-01

    BACKGROUND: Few reports are found with J-1 acellular dermal matrix to prevent postoperative complications after impactedmandibular third molar extraction.OBJECTIVE: To evaluate the effects of acellular dermal matrix embedded in socket after wisdom tooth extraction.METHODS: 400 patients with impacted mandibular third molar were divided into two groups at random with 200 in each group. Ingroup A, the acellular dermal matrix was embedded in the sockets after wisdom tooth extraction; group B was the blank control.Postoperative complicati ons of the two groups were observed after treatment.RESULTS AND CONCLUSION: No acellular dermal matrix lost from wisdom tooth extraction sokets. Blooding after toothextraction decreased when acellular dermal matrix was embedded in the sokets, hemorrhage incidence rate was lower in group Athan in group B. Acellular dermal matrix could prevent blood clot lose from sockets, and also can prevent food residual enteringinto the sockets. Dry socket incidence rate was lower in group A than group B. Acellular embedded in the sockets had no effectson swelling incidence rate. Acellular dermal matrix embedded in socket after tooth extraction can prevent dry socket andhemorrhage, but can not prevent swelling.%背景:使用J-1型脱细胞异体真皮组织补片覆盖拔牙创口的报告较少.目的:探讨异体脱细胞组织补片置入拔牙窝对预防拔牙后并发症的影响.方法:将400例阻生智齿拔除患者随机分为2组,实验组智齿拔除后拔牙窝内放置医用组织补片;对照组智齿拔除后不放置医用组织补片.分别观察拔牙后组织补片脱落率、肿胀发生率、拔牙窝内血凝块存留和食物残渣残留情况、牙龈是否红肿、对拔牙后出血的影响以及干槽症的发生率.结果与结论:拔牙后出血的百分比,血凝块留存率,拔牙窝内食物残渣残留百分比,干槽症发生率实验组均明显低于对照组.放置组织补片对术后

  13. PREPARATION AND BIOCOMPATIBILITY OF PORCINE SKELETAL MUSCLE ACELLULAR MATRIX FOR ADIPOSE TISSUE ENGINEERING%骨骼肌无细胞基质的制备及其生物相容性研究

    Institute of Scientific and Technical Information of China (English)

    田春祥; 范雪娇; 陈晓禾; 邓力; 秦廷武; 罗静聪; 李秀群; 吕青

    2012-01-01

    Objective Extracellular matrix is one of the focus researches of the adipose tissue engineering. To investigate the appropriate method to prepare the porcine skeletal muscle acellular matrix and to evaluate the biocompatibility of the matrix. Methods The fresh skeletal muscle tissues were harvested from healthy adult porcine and were sliced into 2-3 mm thick sheets, which were treated by hypotonic-detergent method to remove the cells from the tissue. The matrix was then examined by histology, immunohistochemistry, and scanning electron microscopy. The toxic effects of the matrix were tested by MTT. Human adipose-derived stem cells (hADSCs) were isolated from adipose tissue donated by patients with breast cancer, and identified by morphology, flow cytometry, and differentiation ability. Then, hADSCs of passage 3 were seeded into the skeletal muscle acellular matrix, and cultured in the medium. The cellular behavior was assessed by calcein-AM (CA) and propidium iodide (PI) staining at 1st, 3rd, 5th, and 7th days after culturing. Results Histology, immunohistochemistry, and scanning electron microscopy showed that the muscle fibers were removed completely with the basement membrane structure; a large number of collagenous matrix presented as regular network, porous-like structure. The cytotoxicity score of the matrix was grade 1, which meant that the matrix had good cytocompatibil ity. The CA and PI staining showed the seeded hADSCs had the potential of spread and proliferation on the matrix. Conclusion Porcine skeletal muscle acellular matrix has good biocompatibility and a potential to be used as an ideal biomaterial scaffold for adipose tissue engineering.%目的 细胞外基质是脂肪组织工程材料的研究热点之一.通过探讨骨骼肌无细胞基质的制作方法及生物相容性,为其在脂肪组织工程中的应用奠定基础. 方法 取健康成年小香猪新鲜骨骼肌组织,横切成厚2~3mm的组织块,采用低渗-去垢剂法脱细胞

  14. Acellular bone marrow extracts significantly enhance engraftment levels of human hematopoietic stem cells in mouse xeno-transplantation models.

    Directory of Open Access Journals (Sweden)

    Kazem Zibara

    Full Text Available Hematopoietic stem cells (HSC derived from cord blood (CB, bone marrow (BM, or mobilized peripheral blood (PBSC can differentiate into multiple lineages such as lymphoid, myeloid, erythroid cells and platelets. The local microenvironment is critical to the differentiation of HSCs and to the preservation of their phenotype in vivo. This microenvironment comprises a physical support supplied by the organ matrix as well as tissue specific cytokines, chemokines and growth factors. We investigated the effects of acellular bovine bone marrow extracts (BME on HSC in vitro and in vivo. We observed a significant increase in the number of myeloid and erythroid colonies in CB mononuclear cells (MNC or CB CD34+ cells cultured in methylcellulose media supplemented with BME. Similarly, in xeno-transplantation experiments, pretreatment with BME during ex-vivo culture of HSCs induced a significant increase in HSC engraftment in vivo. Indeed, we observed both an increase in the number of differentiated myeloid, lymphoid and erythroid cells and an acceleration of engraftment. These results were obtained using CB MNCs, BM MNCs or CD34(+ cells, transplanted in immuno-compromised mice (NOD/SCID or NSG. These findings establish the basis for exploring the use of BME in the expansion of CB HSC prior to HSC Transplantation. This study stresses the importance of the mechanical structure and soluble mediators present in the surrounding niche for the proper activity and differentiation of stem cells.

  15. Acellular nerve allograft promotes selective regeneration

    Institute of Scientific and Technical Information of China (English)

    Haili Xin; Guanjun Wang; Xinrong He; Jiang Peng; Quanyi Guo; Wenjing Xu

    2011-01-01

    Acellular nerve allograft preserves the basilar membrane tube and extracellular matrix, which pro-motes selective regeneration of neural defects via bridging. In the present study, a Sprague Dawley rat sciatic nerve was utilized to prepare acellular nerve allografts through the use of the chemical extraction method. Subsequently, the allograft was transplanted into a 10-mm sciatic nerve defect in Wistar rats, while autologous nerve grafts from Wistar rats served as controls. Compared with autologous nerve grafts, the acellular nerve allografts induced a greater number of degenerated nerve fibers from sural nerves, as well as a reduced misconnect rate in motor fibers, fewer acetyl-choline esterase-positive sural nerves, and a greater number of carbonic anhydrase-positive senso-ry nerve fibers. Results demonstrated that the acellular nerve allograft exhibited significant neural selective regeneration in the process of bridging nerve defects.

  16. Biocompatibility of acellular dermal matrix graft evaluated in culture of murine macrophages Avaliação da biocompatibilidade da matriz dérmica acelular em cultura de células

    Directory of Open Access Journals (Sweden)

    Ana Paula Vendramini

    2006-04-01

    Full Text Available The acellular dermal matrix allograft has been used as an alternative to autogenous palatal mucosal graft. The aim of this study was the evaluation of the biocompatibility of an acellular dermal matrix (AlloDerm® in culture of macrophages. For hydrogen peroxidase determination we used the method of Pick & Kesari, and the Griess method for nitric oxide determination,. Statistical analysis showed no significant difference (p A matrix dérmica acelular tem sido utilizada como alternativa para a substituição de enxerto gengival autógeno. O objetivo deste estudo foi avaliar a biocompatibilidade em cultura de células de macrófagos da matriz dérmica acelular (AlloDermâ. Foram utilizados os métodos de Pick & Kesari, para a determinação da presença de peróxido de hidrogênio (H2O2 e de Griess para a determinação de ácido nitroso (NO. Não houve diferença estatisticamente significante (p < 0,05 no aumento da presença de NO e H2O2 quando macrófagos foram expostos na presença da matrix dérmica acelular quando comparado com o controle negativo. Pode-se concluir que a matrix dérmica acelular é biocompatível aos tecidos humanos.

  17. Acellular Lung Scaffolds Direct Differentiation of Endoderm to Functional Airway Epithelial Cells: Requirement of Matrix-Bound HS Proteoglycans

    Directory of Open Access Journals (Sweden)

    Sharareh Shojaie

    2015-03-01

    Full Text Available Efficient differentiation of pluripotent cells to proximal and distal lung epithelial cell populations remains a challenging task. The 3D extracellular matrix (ECM scaffold is a key component that regulates the interaction of secreted factors with cells during development by often binding to and limiting their diffusion within local gradients. Here we examined the role of the lung ECM in differentiation of pluripotent cells in vitro and demonstrate the robust inductive capacity of the native lung matrix alone. Extended culture of stem cell-derived definitive endoderm on decellularized lung scaffolds in defined, serum-free medium resulted in differentiation into mature airway epithelia, complete with ciliated cells, club cells, and basal cells with morphological and functional similarities to native airways. Heparitinase I, but not chondroitinase ABC, treatment of scaffolds revealed that the differentiation achieved is dependent on heparan sulfate proteoglycans and its bound factors remaining on decellularized scaffolds.

  18. Acellular organ scaffolds for tumor tissue engineering

    Science.gov (United States)

    Guller, Anna; Trusova, Inna; Petersen, Elena; Shekhter, Anatoly; Kurkov, Alexander; Qian, Yi; Zvyagin, Andrei

    2015-12-01

    Rationale: Tissue engineering (TE) is an emerging alternative approach to create models of human malignant tumors for experimental oncology, personalized medicine and drug discovery studies. Being the bottom-up strategy, TE provides an opportunity to control and explore the role of every component of the model system, including cellular populations, supportive scaffolds and signalling molecules. Objectives: As an initial step to create a new ex vivo TE model of cancer, we optimized protocols to obtain organ-specific acellular matrices and evaluated their potential as TE scaffolds for culture of normal and tumor cells. Methods and results: Effective decellularization of animals' kidneys, ureter, lungs, heart, and liver has been achieved by detergent-based processing. The obtained scaffolds demonstrated biocompatibility and growthsupporting potential in combination with normal (Vero, MDCK) and tumor cell lines (C26, B16). Acellular scaffolds and TE constructs have been characterized and compared with morphological methods. Conclusions: The proposed methodology allows creation of sustainable 3D tumor TE constructs to explore the role of organ-specific cell-matrix interaction in tumorigenesis.

  19. 异种(猪)无细胞真皮基质的制备及体外生物相容性%Preparation and in vitro biocompatibility of xenogenic(porcine)acellular dermal matrix

    Institute of Scientific and Technical Information of China (English)

    马绍英; 李宝明; 董丽; 王旭昇; 李宝兴; 赵亚平; 康悦

    2009-01-01

    背景:人同种无细胞真皮基质作为一种永久性真皮支架,已成功应用于烧伤创面修复及美容医学等领域,但由于来源有限,限制了其应用.目的;研制异种(猪)无细胞真皮基质,并对其体外生物相容性进行评价.设计、时间及地点:体外细胞学对比观察实验,于2007-08/2008-06在中国辐射防护研究院生物材料与制药技术研究所实验室完成.材料:实验猪由中国辐射防护研究院实验动物中心提供;人成纤维细胞来自武警山西总队医院健康儿童包皮环切术切除的包皮组织.方法:无菌条件下获取健康小白猪猪皮,用高渗盐溶液-去污剂、胰酶消化及超声清洗的方法,制备猪无细胞真皮基质.体外培养人成纤维细胞,用猪无细胞真皮基质浸提液法及人成纤维细胞和猪无细胞真皮基质直接贴附法,评价猪无细胞真皮基质体外生物相容性.主要观察指标:①猪无细胞真皮基质的组织学形态.②猪无细胞真皮基质的体外生物相容性.结果:制备的猪无细胞真皮基质,完全去除了表皮和真皮中的细胞成分,保留了胶原基质.猪无细胞真皮基质浸提液对人成纤维细胞增殖无明显影响.人成纤维细胞可以在猪无细胞真皮基质上贴附、增殖.结论:此种方法制备的无细胞真皮基质完全去除了表皮和真皮中的细胞成分,有较好的体外生物相容性.%BACKGROUND: Human allogenic acellular dermal matrix, as a kind of permanent dermal scaffold, has widely used in the fields of burn wound reparation and aesthetic medicine. However, it is limited due to insufficient resources. OBJECTIVE: To prepare porcine acellular dermal matrix (PADM) dermal matrix, in addition, to estimate its in vitro biocompatibility. DESIGN, TIME AND SETTING: An in vitro cytology contrast experiment. The Experiment was performed at the laboratory of Biomaterials and Pharmacy Technology Institute, China Institute for Radiation Protection

  20. A nanomedicine approach to effectively inhibit contracture during bladder acellular matrix allograft-induced bladder regeneration by sustained delivery of vascular endothelial growth factor.

    Science.gov (United States)

    Xiong, Qianwei; Lin, Houwei; Hua, Xiaolin; Liu, Li; Sun, Ping; Zhao, Zhen; Shen, Xiaowei; Cui, Daxiang; Xu, Maosheng; Chen, Fang; Geng, Hongquan

    2015-01-01

    Macroscopic evidence of contracture has been identified as a major issue during the regeneration process. We hypothesize that lack of angiogenesis is the primary cause of contracture and explore a nanomedicine approach to achieve sustained release of vascular endothelial growth factor (VEGF) to stimulate angiogenesis. We evaluate the efficacy of poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) for long-term (3 months) sustained release of VEGF in bladder acellular matrix allografts (BAMA) in a swine model. We anticipate that the sustained release of VEGF could stimulate angiogenesis along the regeneration process and thereby inhibit contracture. Bladder was replaced with BAMA (5×5 cm), modified with PLGA NPs encapsulated with VEGF in a pig model. The time points chosen for sampling were 1, 2, 4, and 12 weeks. The regenerated areas were then measured to obtain the contracture rate, and the extent of revascularization was calculated using histological and morphological features. In the control group of animals, the bladder was replaced with only BAMA. The in vivo release of VEGF was evident for ∼3 months, achieving the goal of long-acting sustained release, and successfully promoted the regeneration of blood vessels and smooth muscle fibers. In addition, less collagen deposition was observed in the experimental group compared with control. Most importantly, the inhibition of contracture was highly significant, and the ultimate contracture rate decreased by ∼57% in the experimental group compared with control. In isolated strips analysis, there were no significant differences between BAMA-regenerated (either VEGF added or not) and autogenous bladder. BAMA modified with VEGF-loaded PLGA-NPs can sustainably release VEGF in vivo (>3 months) to stimulate angiogenesis leading to the inhibition of contracture. This is the first study to report a viable nanomedicine-based strategy to overcome contracture during bladder regeneration induced by BAMA. Furthermore

  1. Early Surgical Site Infection Following Tissue Expander Breast Reconstruction with or without Acellular Dermal Matrix: National Benchmarking Using National Surgical Quality Improvement Program

    Directory of Open Access Journals (Sweden)

    Sebastian Winocour

    2015-03-01

    Full Text Available BackgroundSurgical site infections (SSIs result in significant patient morbidity following immediate tissue expander breast reconstruction (ITEBR. This study determined a single institution's 30-day SSI rate and benchmarked it against that among national institutions participating in the American College of Surgeons National Surgical Quality Improvement Program (ACS-NSQIP.MethodsWomen who underwent ITEBR with/without acellular dermal matrix (ADM were identified using the ACS-NSQIP database between 2005 and 2011. Patient characteristics associated with the 30-day SSI rate were determined, and differences in rates between our institution and the national database were assessed.Results12,163 patients underwent ITEBR, including 263 at our institution. SSIs occurred in 416 (3.4% patients nationwide excluding our institution, with lower rates observed at our institution (1.9%. Nationwide, SSIs were significantly more common in ITEBR patients with ADM (4.5% compared to non-ADM patients (3.2%, P=0.005, and this trend was observed at our institution (2.1% vs. 1.6%, P=1.00. A multivariable analysis of all institutions identified age ≥50 years (odds ratio [OR], 1.4; confidence interval [CI], 1.1-1.7, body mass index ≥30 kg/m2 vs. 4.25 hours (OR, 1.9; CI, 1.5-2.4 as risk factors for SSIs. Our institutional SSI rate was lower than the nationwide rate (OR, 0.4; CI, 0.2-1.1, although this difference was not statistically significant (P=0.07.ConclusionsThe 30-day SSI rate at our institution in patients who underwent ITEBR was lower than the nation. SSIs occurred more frequently in procedures involving ADM both nationally and at our institution.

  2. "High-grade" central acellular carcinoma and matrix-producing carcinoma of the breast: correlation between ultrasonographic findings and pathological features.

    Science.gov (United States)

    Yamaguchi, Rin; Tanaka, Maki; Mizushima, Yasuko; Hirai, Yoshitake; Yamaguchi, Miki; Terasaki, Hiroshi; Yokoyama, Toshiro; Tsuchiya, Shin-ichi; Nakashima, Osamu; Yano, Hirohisa

    2011-09-01

    High-grade carcinoma with a large central acellular zone (central acellular carcinoma, CAC) and matrixproducing carcinoma (MPC) are aggressive tumors that both have a central myxomatous acellular zone. Their characteristic morphology may be useful in diagnostic imaging. Ultrasonographic findings based on the Breast Imaging Recording and Data System (BI-RADS) and detailed histological features were evaluated in 11 cases of CAC and 2 cases of MPC to characterize their features. Safranin-O staining was undertaken for the evaluation of central acellular zones in these tumors. Overall, ultrasonography demonstrated heterogeneous hyperechoic lesions in the center of the hypoechoic mass. Posterior echo enhancement was observed in all but 1 case. One case was classified as malignant and the others as "borderline." Histologically, cancer tissue was located in the periphery of the tumor with a ring-like structure and fewer cellular central areas comprising hyaline cartilage myxoid material such as those stained by safranin-O. The present study showed that the pathological findings of CACs and MPCs accurately reflect the ultrasonographic findings. Tumors that showed hyperechoic areas in the center of the hypoechoic mass, with posterior echo enhancement indicating acellular zones composed by myxochondroid material, and that were also relatively round on ultrasonography may be benign, but evaluation is required to exclude CAC and MPC.

  3. 交联剂对脱细胞膀胱基质生物力学性能的影响%Effects of different crosslinking agents on biomechanical properties of acellular bladder matrix

    Institute of Scientific and Technical Information of China (English)

    范雪梅; 李胜平; 徐惠成

    2013-01-01

    目的 研究不同交联剂对猪脱细胞膀胱基质的组织结构影响,并比较其生物力学性能,为盆底修复替代材料的选择提供依据.方法 采用表面活性剂+酶消化法去除新鲜猪膀胱的细胞成分,将脱细胞膀胱基质随机分为3组,A组经0.25%戊二醛交联,B组经0.625%京尼平交联,C组未交联.对各组材料进行HE染色,观察纤维的变化情况.使用生物力学性能测试系统检测抗拉强度、断裂伸长率、弹性模量,并进行统计学分析.结果 京尼平交联脱细胞膀胱基质后呈深蓝色,保持了天然组织构架的完整,纤维更加致密.戊二醛交联脱细胞膀胱基质后呈浅黄色,纤维排列紊乱且有断裂.新鲜猪膀胱经上述三种方法处理后,其弹性模量增大、断裂伸长率减小,而其中京尼平交联处理的脱细胞膀胱基质力学性能与新鲜膀胱组织更为相近.结论 京尼平交联的脱细胞膀胱基质组织结构的形态佳,同时较大限度地保留膀胱组织的力学性能,可能是较理想的盆底重建材料.%Objective To compare the effects of different crosslinking agents on structures and biomechanical properties of porcine acellular bladder matrix.This could provide the basic selection of ideal biomaterials for the reconstruction of female pelvis.Methods Cellular components of fresh porcine bladder were removed by detergent-enzymatic method.They were randomly divided into three groups,and group A was crosslinked with 0.25%glutaraldehyde,group B was crosslinked with 0.625% genipin,group C wasn't crosslinked.Then all samples were examined the morphology with hematoxylin and eosin staining,and biomechanical tests were also performed and evaluated the biomechanical properties by statistical analysis.Results The genipin-crosslinked acellular bladder matrix was dark blue,with very well preserved collagen fiber.The glutaraldehyde-crosslinked acellular bladder matrix was light yellow,with disordered and fracture

  4. 灌注法制备肺脱细胞基质%Pulmonary perfusion for preparation of the acellular matrix

    Institute of Scientific and Technical Information of China (English)

    龚正

    2015-01-01

    背景:对组织器官进行脱细胞,细胞外基质在经过去除细胞和可溶性蛋白处理之后,可维持正常的器官外形及组织结构基质成分.目的:利用灌注法制备肺脱细胞基质.方法:将 40 只 Wistar 大鼠随机均分为 2 组,常规麻醉处理后打开胸腔,获得完整的肺组织,实验组利用Langendorff灌注模型构建大鼠全肺脱细胞基质支架,对照组不予以特殊处理.动态观察和记录实验组肺脏颜色和形态变化,分别从 2 组大鼠肺脏不同部位获取小块组织,利用电子显微镜进行组织学观察,观察两组的Weigert弹力纤维联合Von Gieson结缔组织染色和苏木精-伊红染色情况.结果与结论:经1%脱氧胆酸钠溶液灌注后,实验组大鼠肺脏颜色逐渐发生改变,表现为从内至外呈分段分叶状逐渐变为白色半透明状,并可观察到清晰肺叶结构,最终肺脏呈现出均一的白色半透明状.经 Weigert 弹力纤维联合Von Gieson结缔组织染色和苏木精-伊红染色发现,对照组新鲜肺组织切片中含有大量细胞,其中包括毛细血管和成纤维细胞及内皮细胞等,细胞呈整齐排列状,具有完整的肺泡结构,弹性纤维结构清晰,胶原纤维也呈整齐排列状,结构较为紧凑;实验组肺组织细胞基本已消失,仍具有完整的肺泡形态结构,但呈较为疏松的状态且裂隙增大,弹性纤维保存良好,胶原纤维呈较疏松排列状.表明利用灌注法课可有效构建大鼠全肺组织基质支架.%BACKGROUND:The extracelular matrix with removal of cels and soluble proteins can maintain the normal shape of organs and matrix components. OBJECTIVE:To prepare the acelular matrix of lung tissue using perfusion method. METHODS:Forty Wistar rats were randomly divided into two groups: under routine anesthesia, the chest was open to obtain complete lung tissue for construction of rat lung acelular matrix scaffold using Langendorff perfusion model in experimental group

  5. Application effect comparison on allogenic acellular dermal matrix and xenogenic acellular dermal matrix in the treatment of burn wounds%异体脱细胞真皮和异种脱细胞真皮在烧伤创面治疗中的应用效果比较

    Institute of Scientific and Technical Information of China (English)

    彭炳生; 柳晖

    2015-01-01

    Objective To compare the effect on allogenic acellular dermal matrix and xenogenic (pig) acellular dermal matrix in the treatment of burn wounds, to explore the feasibility of xenogeneic acellular dermal substitute acellular dermal. Methods In Bao’an People’s Hospital of Shenzhen City from May 2012 to June 2013 with deep second degree and third degree burn wounds of deep partial tangential excision and skin grafting for treatment of 30 patients of autologous skin at the same time, they were divided into two groups by the random digital table method, allograft +micro skin treatment were used in allograft group of 15 patients, xenogenic (pig) acellular dermal matrix + micro skin treatment were applied in heterogeneous group of 15 patients. The wound healing time, the healing quality between two groups were compared; the patients were followed up for 6 months, the self satisfaction of Vancouver scar scale and patient evaluation table were used to evaluate the healing effect. Results The short-term effect:there were 16 (51.61%) one type of healing and 15 (48.39%) two type of healing in 31 wounds of allograft group, there were 15 (50.00%) one type of healing and 15 (50.00%) two type of healing in 30 wounds of heterogeneous group, with no statistical difference (P>0.05). Drying time of wound healing, deep second degree burn wound healing time and third degree burn wound healing time between two groups was compared respectively, there was no statistical difference (P > 0.05). The long term effect: after 6 months of follow-up, color, blood vessel distribution, thickness and softness scores between two groups had no statistical difference (P>0.05). The excellent and good rate of allograft group and heterogeneous group was 51.61%and 50.00% respectively, with no statistical difference (P>0.05). Conclusion The curative effects of two materials are similar, can replace each other, its clinical application can alleviate allograft skin source, at the same time, reduce

  6. A single-arm trial indirect comparison investigation: a proof-of-concept method to predict venous leg ulcer healing time for a new acellular synthetic matrix matched to standard care control.

    Science.gov (United States)

    Shannon, Ronald; Nelson, Andrea

    2017-08-01

    To compare data on time to healing from two separate cohorts: one treated with a new acellular synthetic matrix plus standard care (SC) and one matched from four large UK pragmatic, randomised controlled trials [venous leg ulcer (VLU) evidence network]. We introduce a new proof-of-concept strategy to a VLU clinical evidence network, propensity score matching and sensitivity analysis to predict the feasibility of the new acellular synthetic matrix plus SC for success in future randomised, controlled clinical trials. Prospective data on chronic VLUs from a safety and effectiveness study on an acellular synthetic matrix conducted in one wound centre in the UK (17 patients) and three wound centres in Australia (36 patients) were compared retrospectively to propensity score-matched data from patients with comparable leg ulcer disease aetiology, age, baseline ulcer area, ulcer duration, multi-layer compression bandaging and majority of care completed in specialist wound centres (average of 1 visit per week), with the outcome measures at comparable follow-up periods from patients enrolled in four prospective, multicentre, pragmatic, randomised studies of venous ulcers in the UK (the comparison group; VLU evidence network). Analysis using Kaplan-Meier survival curves showed a mean healing time of 73·1 days for ASM plus SC (ASM) treated ulcers in comparison with 83·5 days for comparison group ulcers treated with SC alone (Log rank test, χ(2) 5·779, P = 0·016) within 12 weeks. Sensitivity analysis indicates that an unobserved covariate would have to change the odds of healing for SC by a factor of 1·1 to impact the baseline results. Results from this study predict a significant effect on healing time when using a new ASM as an adjunct to SC in the treatment of non-healing venous ulcers in the UK, but results are sensitive to unobserved covariates that may be important in healing time comparison. © 2016 Medicalhelplines.com Inc and John Wiley & Sons Ltd.

  7. In vivo effects of human adipose-derived stem cells reseeding on acellular bovine pericardium in nude mice.

    Science.gov (United States)

    Wu, Qingkai; Dai, Miao; Xu, Peirong; Hou, Min; Teng, Yincheng; Feng, Jie

    2016-01-01

    Tissue-engineered biologic products may be a viable option in the reconstruction of pelvic organ prolapse (POP). This study was based on the hypothesis that human adipose-derived stem cells (hASCs) are viable in acellular bovine pericardium (ABP), when reseeded by two different techniques, and thus, aid in the reconstruction. To investigate the reseeding of hASCs on ABP grafts by using non-invasive bioluminescence imaging (BLI), and to identify the effective hASCs-scaffold combinations that enabled regeneration. Thirty female athymic nude mice were randomly divided into three groups: In the VIVO group, ABPs were implanted in the subcutaneous pockets and enhanced green fluorescent protein luciferase (eGFP·Luc)-hASCs (1 × 10(6) cells/50 µL) were injected on the ABP at the same time. In the VITRO group, the mice were implanted with grafts that ABP were co-cultured with eGFP·Luc-hASCs in vitro. The BLANK group mice were implanted with ABP only. The eGFP·Luc-hASCs reseeded on ABP were analyzed by BLI, histology, and immunohistochemistry. The eGFP·Luc-hASCs reseeded on ABP could be visualized at 12 weeks in vivo. Histology revealed that the VIVO group displayed the highest cell ingrowths, small vessels, and percent of collagen content per unit area. Desmin and α-smooth muscle actin were positive at the same site in the VIVO group cells. However, few smooth muscles were observed in the VITRO and BLANK groups. These results suggest that hASCs reseeded on ABP in vivo during surgery may further enhance the properties of ABP and may promote regeneration at the recipient site, resulting in a promising treatment option for POP. © 2016 by the Society for Experimental Biology and Medicine.

  8. Antibacterial activity of acellular dermal matrix composite CMC%复合 CMC 脱细胞真皮基质的抗菌性能

    Institute of Scientific and Technical Information of China (English)

    安祥莲; 郭泾

    2015-01-01

    目的:探讨脱细胞真皮基质(ADM)以戊二醛为交联剂复合羧甲基壳聚糖(CMC)后的抗菌性能。方法实验采用2×3析因设计,根据样本状态(湿润、干燥)和分组(复合样片组、单纯样片组、空白对照组)设计实验。应用烧瓶振荡法,利用优化的实验条件,在转速200 r/min、培养温度25℃、作用时间1 h 的条件下,对复合 CMC 的ADM进行抗菌性能检测。通过菌落平板计数获得实验前(0 h)、实验后(1 h)的菌落数,计算抑菌率。两组实验中细菌在振摇前后的菌落数差值均符合实验限定标准(差值变化不超过10%)。结果湿润状态下,复合样片组对金黄色葡萄球菌的抑菌率为61.17%,对白色念珠菌的抑菌率为39.79%;干燥状态下,复合样片组对金黄色葡萄球菌的抑菌率为0%,对白色念珠菌的抑菌率为15.17%。湿润与干燥两个因素比较,差异有统计学意义(P <0.001)。结论复合 CMC 的 ADM 这种交联复合物在湿润状态下有一定的抗菌性,干燥状态下不具备抗菌性能,交联效率较低,需进一步改良制备工艺。%Objective To explore the antibacterial activity of acellular dermal matrix (ADM)with glutaraldehyde as crosslinking agent composite carboxymethyl chitosan(CMC).Methods The 2 ×3 factorial design was adopted,and the experiments were designed according to the sample status (wet,dry)and groups (composite sample group,simple sample group,blank control group).Shaking-flask method was used and the optimized experimental conditions (rota-tional speed 200 r/min,culture temperature 25 ℃,culture time one hour)were used to evaluate the antibacterial activity of acellular dermal matrix composite CMC.To get the bacterial inhibition rate,colony plate count was adopted to obtain the number of bacterial colonies before the experiment(0 h)and after the test(1 h).In the two experimental groups, the number

  9. Feasibility of eyelid reconstruction with acellular xenogenic dermal matrix%异种脱细胞真皮替代睑板材料重建眼睑的可行性

    Institute of Scientific and Technical Information of China (English)

    张向荣; 周琼; 肖卫; 刘德伍; 彭燕

    2011-01-01

    背景:眼睑后层重建是眼睑重建的重点和难点,其中睑板替代物更是研究的焦点.异种脱细胞真皮作为一种新型的组织工程材料,在国内外烧伤整形领域,正得到广泛的研究和应用.目的:观察异种(猪)脱细胞真皮植入兔眼睑后的组织相容性极其组织病理学变化.方法:剥取健康小白猪全层皮肤20 cm×20 cm,制备异种(猪)脱细胞真皮基质.同时制备兔睑板全层缺损模型并植入脱细胞真皮基质,观察大体情况,并分别于第1,2,3周取移植交界处眼睑组织光镜下观察组织学的改变.结果与结论:大体观察未见明显排斥反应及眼睑的变形;光镜下1周时可见局部炎症细胞浸润,2周时炎症细胞减少,3周时正常纤维组织长入,逐渐分割代替植入的胶原纤维,炎症反应消失.提示异种脱细胞真皮免疫原性低,并可引导新生胶原的生长,是一种良好的睑板替代物.%BACKGROUND:Reconstruction of posterior lamella of eyelid is an important and difficult issue in eyelid reconstruction, and tarsal substitute is the focus of the study. Xenogenic acellular dermal matrix as a new tissue engineering material, is being widely studied and applied in the field of burn and plastic su rgery at home and abroad.OBJECTIVE:To observe histocompatibility and histopathological changes of xenogenic (porcine) acellular dermal matrix transplantation for eyelid reconstruction in rabbits.OBJECTIVE:Full skin at 20 cm×20 cm was stripped from healthy little white pigs, for preparation of xenogenic (porcine) acellular xenogenic dermal matrix. Model of rabbit tarsal defect was established in 8 New Zealand rabbits, then acellular dermal matrix was implanted.Samples of implanted materials were collected for histological examination at 1, 2, 3 weeks postoperation under light microscopy.RESULTS AND CONCLUSION:There were no obvious rejection and eyelid deformation observed. One week after implantation,partial inflammatory cells

  10. A comparative evaluation of the effectiveness of subpedicle acellular dermal matrix allograft with subepithelial connective tissue graft in the treatment of isolated marginal tissue recession: A clinical study

    OpenAIRE

    Shori, Tony; Kolte, Abhay; Kher, Vishal; Dharamthok, Swarup; Shrirao, Tushar

    2013-01-01

    Introduction: The most common problem encountered in our day to day practice is exposed root surface or a tooth getting long. The main indication for root coverage procedures are esthetics and/or cosmetic demands followed by the management of root hypersensitivity, root caries or when it hampers proper plaque removal. Over the years, various techniques have been used to achieve root coverage. Aim and Objectives: The aim of this study was to compare the effectiveness of subpedicle acellular de...

  11. 同种异体脱细胞真皮用于HA义眼台暴露修复观察%Clinical observation of using homologous acellular dermal matrix as repairing material in the exposure of orbital hydroxyapatite implants

    Institute of Scientific and Technical Information of China (English)

    王超庆; 李燕飞; 程秀春; 李琦; 范晓聪; 李静

    2015-01-01

    Objective To understand the security and effectiveness of using homologous acellular dermal as repairing material in the exposure of orbital hydroxyapatite (HA) implants.Methods By 13 cases (13 eyes) hydroxyapatite implant exposure with intraoperative exploration,to initially judge implantation methods and exposure reasons of orbital implants,and using homologous acellular dermal as repairing material.Results Thirteen cases of postoperative conjunctival blood supply were good,no obvious edema and exudation,no incision dehiscence,infection and other complications.In continuous observation of 10 months,no evidence of exposure recurred.Conclusions Acellular dermal material is readily available,and has good tissue compatibility,resistance to infection,certain flexibility,and the immune response induced is light.Acellular dermal as a extracellular matrix can guide the organization's own collagen fibers ingrowth,and is easy to survive.For the repair of HA orbital implants after implant exposure is a good material.%目的 了解同种异体脱细胞真皮应用于羟基磷灰石(HA)义眼台植入术后暴露的修复治疗的有效性和安全性.方法 对2010~2013年在济南市明水眼科医院就诊的13例(13只眼)HA义眼台暴露患者术中探查,初步判断HA义眼台植入术式及暴露的原因,应用脱细胞真皮覆盖的方法进行处理.结果 13例患者术后结膜血运良好,无明显水肿及渗出,无切口哆开、感染等并发症.连续观察10个月,未见暴露复发迹象.结论 脱细胞真皮材料容易获得,组织相容性好,抗感染能力强,有一定的柔韧性,引起的免疫反应轻微,作为一种细胞外基质,可引导自身组织的胶原纤维长入,对创面要求低易于成活.对修复HA义眼台植入术后眼台暴露是一种较好的材料.

  12. 脱细胞基质补片在腹股沟疝修补术中的应用%Acellular tissue matrix mesh for inguinal herniopasty

    Institute of Scientific and Technical Information of China (English)

    李绍杰; 唐健雄; 黄磊; 蔡昭; 陈革

    2011-01-01

    Objective To evaluate acellular tissue matrix mesh in inguinal hernioplasty in young male patients.Methods Clinical outcome and sexual function were evaluated on 12 young male inguinal hernia patients undergoing inguinal hernioplasty by ACTM mesh.Results The mean operation time was 62 minutes.Postoperative analgesics were needed for an average of 1.45 days (range,0 -4 days).The average postoperative hospital stay was 3.9 days,and patients were able to return to normal activity,after a mean 11.8 days.All patients were followed-up for a mean duration of 9.54 months(range,6 - 16 months).Two patients complained pain one month after surgery and the pain disappeared later without any medical treatment.There was no hernia recurrence and foreign body feelings,nor sexual and reproductive problems.Conclusions The ACTM mesh is a safe and effective material for inguinal hernioplasty,especially in young male patients.%目的 研究脱细胞基质生物补片(ACTM)在中青年男性腹股沟疝修补术中的应用价值.方法 采用脱细胞基质生物补片(ACTM)对12例中青年男性单侧腹股沟疝患者实施腹股沟疝无张力修补术,观察其临床疗效、副反应以及对患者生殖情况的影响.结果 12例患者平均手术时间(62±18) min,服用止痛药时间0~4d,平均(1.4±0.8)d.出院时间为术后2~5d,平均(3.9±0.4)d.12例患者恢复日常工作及活动时间为术后5~21 d,平均(11.8±2.5)d.12例患者术后随访6~16个月,平均(9.5±1.3)个月.其中2例(2/12,18.3%)在术后1个月有疼痛不适感,未经特殊处理,术后3个月疼痛均好转.所有患者无异物感及局部积液等表现,均无性功能障碍.结论 脱细胞基质生物补片用于腹股沟疝修补术安全有效,无严重的短期并发症,适用于有生育要求的中青年男性患者.

  13. Guide bone regeneration with acellular dermal matrix in the maxillary anterior region%脱细胞真皮基质在上前牙GBR种植术中的临床研究

    Institute of Scientific and Technical Information of China (English)

    董强; 夏茜; 马洪; 王小玲; 杨红; 周成菊; 毛久凤

    2014-01-01

    目的:研究采用脱细胞真皮基质进行引导骨组织再生技术(GBR)并同期种植体植入的短期临床效果。方法29例上前牙脱细胞真皮基质进行 GBR并同期植入种植体,经软组织塑形后,完成最终上部结构。随访3~9个月,对种植体周围软硬组织进行评价。结果29例均获得良好骨整合,种植体无松动脱落。种植体周围软硬组织状态良好,美学效果满意。结论采用脱细胞真皮基质进行上前牙 GBR并同期植入种植体,短期内可获得较满意的临床效果。%Objective To evaluate the clinical and aesthetic results of guide bone regeneration(GBR)with acellular dermal matrix and implant placement in the maxillary anterior region.Methods 29 cases in the maxillary anterior region were selected carefully, GBR with acellular dermal matrix were processed and the implants were placed immediately.Impressions were taken after soft tis-sue development with provisional implant restorations and the definitive restorations were finished.The follow-up time was 3 to 9 months.The evaluated indexes involved marginal bone level at mesial and distal aspects of the implants,the interproximal papilla in-dex score of Jemt′s classification and the level of the labial soft-tissue margin.Results 29 cases were good bone integration,implant without mobility.Hard and soft tissue around implants in good condition,aesthetic effect was satisfied.Conclusion Using acellular dermal matrix to come forward to tooth GBR and implanted implant during this period,can obtain satisfactory clinical effect in the short term.

  14. Aplicación clínica de la matriz dérmica acelular para prevenir recesiones gingivales Clinical application of acellular dermal matrix to prevent gingival recessions

    Directory of Open Access Journals (Sweden)

    C.M. Ardila Medina

    2009-04-01

    Full Text Available Un objetivo primordial de la cirugía plástica periodontal es cubrir las superficies radiculares expuestas cuando esta condición causa al paciente problemas estéticos, hipersensibilidad dentinal, caries radicular o dificulta una adecuada remoción de la placa bacteriana. Muchas técnicas quirúrgicas se han propuesto para la corrección de exposiciones radiculares: autoinjerto gingival libre, injertos pediculados y técnicas bilaminares. La regeneración tisular guiada también se ha ofrecido como otra alternativa terapéutica en el manejo de recesiones gingivales. Un aloinjerto de matriz dérmica acelular (AMDA se ha reportado recientemente en la literatura periodontal, mostrando resultados clínicos favorables en el cubrimiento de recesiones gingivales. El objetivo de esta revisión es mostrar la composición del AMDA, sus características, antecedentes y predecibilidad comparado con otras técnicas para lograr cubrimiento de recesiones gingivales.The ultimate goal of periodontal surgery is the coverage of exposed root surface when this condition causes the patient esthetic troubles, dentinal hipersentivity, or root caries or when it hampers proper plaque removal. Many surgical techniques have been proposed for the correction of dental root exposition: free gingival grafts, pedicle soft tissue grafts and bilaminar techniques. Guided tissue regeneration has also been proposed as a possible therapeutic alternative in the management of gingival recession. Recently, an acellular dermal matrix allograft (ADMA has been reported to have a favorable clinical outcome in coverage of gingival recessions. The objective of this review is to show composition, qualitys, trajectory and mainly predictable of the acellular dermal matrix allograft to compare with others techniques to cover gingival recessions.

  15. 猪脱细胞真皮基质修复兔腹壁缺损的实验研究%PORCINE ACELLULAR DERMAL MATRIX FOR REPAIR OF ABDOMINAL WALL DEFECTS IN RABBIT MODEL

    Institute of Scientific and Technical Information of China (English)

    马绍英; 王旭昇; 董丽; 李幼忱; 周沫; 赵亚平; 李宝兴

    2011-01-01

    matrix in the reconstruction of abdominal wall defects in rabbits, and to investigate the application feasibility of xeno-transplantation of acellular dermal matrix. Methods The porcine acellular dermal matrix was prepared from a health white pig. Twenty-six Japanese white rabbits (weighing 2.2-2.3 kg, female or male) were randomly assigned to 2 groups: the control group (n=6) and the experimental group (?=20). In the control group, the full-thickness abdominal wall defect of 5.0 cm × 0.5 cm was made, and the defect was sutured directly, in the experimental group, the full-thickness abdominal wall defect of 5.0 cm × 2.5 cm was made, and the defect was repaired with porcine acellular dermal matrix patch at the same size as the defect. At 5 weeks after surgery, the incidence of hernia and the intra-abdominal adhesions were observed and the wound breaking strength was compared between the patch-fascia interface and the fascia-fascia interface. The graft vascularization was evaluated through histological analysis at 6 months after surgery in the experimental group. Results No hernia occurred in all rabbits of 2 groups. At 5 weeks after surgery, healing was observed between patch and the muscularfascia; the vascularization was seen in the porcine acellular dermal matrix patch. There was no significant difference in the adhesion grade (Z= -0.798, P=0.425) between the experimental group (grade 2 in 1 rabbit, grade 1 in 5, and grade 0 in 12) and the control group (grade 1 in 1 and grade 0 in 5). No significant difference was found (t= -0.410, P=0.683) in the breaking strength between the patch-fascia interface in the experimental group [(13.0 ± 5.5) N] and the fascia-fascia interface in control group [(13.6 ± 4.0) N]. In the experimental group, the small vessels and the infiltration of inflammatory cells were observed in the porcine acellular dermal matrix patch after 5 weeks through histological observations. The junctions of the patch-fascia interface healed with fibrous

  16. Clinical application of heterogeneous acellular dermal matrix used in alveolar bone grafting%异种脱细胞真皮基质膜在齿槽裂植骨术中的临床应用

    Institute of Scientific and Technical Information of China (English)

    李可兴; 刘曙光

    2014-01-01

    Objective To observe the effect of bone repair and evaluate its esthetic outcome with heterogeneous acellular dermal matrix cover the alveolar cleft bone grafting area in the alveolar cleft operation.Methods In 67 cases,unilateral cleft palate,were treated by alveolar cleft conventional surgical method.Cancellous iliac bone grafting were control group,heterogeneous acellular dermal matrix cover the alveolar cleft bone grafting area were treatment group.Radiographs was taken at 1 st,3 rd,6 th,12 th,18 th,24 th month postoperatively to observe the bone regeneration alveolar cleft zone.Results The alveolar cleft graft area new bone formation with Ⅰ,Ⅱ,Ⅲ,Ⅳ grade after 6 months in control group was 15,11,9,6 cases and in treatment group was 13,9,3,1 case.The graft survival rate and success rate (97.8%,84.3%) of treatment group were higher than that of control group (84.5%,63.7%),the difference was statistically significant (P < 0.05).Conclusion The successful rate of operation could be warranted,by the joint application of autogenous iliac bone grafts and heterogeneous acellular dermal matrix in the alveolar cleft operation.%目的 在齿槽裂手术中,将异种脱细胞真皮基质膜覆盖齿槽裂植骨区,观察新骨形成状况,评价植骨修复效果.方法 选择67例单侧齿槽裂患者,按治疗方法不同分为对照组和试验组.对照组单纯应用髂骨骨松质移植41例,试验组应用异种脱细胞真皮基质膜加髂骨骨松质移植26例.术后1,3,6,12,18,24个月随访,X线片观察齿槽裂植骨区新骨生成情况.结果 对照组病例术后6个月齿槽裂植骨区新骨形成Ⅰ,Ⅱ,Ⅲ,Ⅳ级分别为15,11,9,6例,齿槽裂植骨成活率为84.5%,临床成功率为63.7%.而试验组病例植骨区新骨形成Ⅰ,Ⅱ,Ⅲ,Ⅳ级分别为13,9,3,1例.齿槽裂植骨成活率为97.8%,临床成功率为84.3%.两组植骨成活率、临床成功率比较差异有统计学意义(P<0.05).结论 自体髂骨加异

  17. Clinical Study of Heterogeneous Acellular Dermal Matrix for Repairing Hard Palatal Fistula%异种脱细胞真皮基质整复硬腭部腭瘘的临床研究

    Institute of Scientific and Technical Information of China (English)

    张波; 李健

    2013-01-01

    [Objective]To explore the clinical efficacy of heterogeneous acellular dermal matrix( Heal-All Oral Biofilm) for repairing hard palatal fistula. [Methods]Thirty-eight patients with hard palate fistula were randomly divided into treatment group and control group. Palatal fistulas of patients in treatment group( n = 18) was repaired with heterogeneous acellular dermal matrix. Palatal fistulas of patients in control group( n = 20) was repaired with traditional method. The recurrence in the closed fistula and the incidence of buccal gingi-val sulcus becoming shallow in patients of two groups were observed 1~3 months after the operation. [Results]The recurrence rate of closed fistula in treatment group was 11. 1 % (2/18) 1 ~ 3 months after the operation, which was markedly lower than that in control group(25. 0%, 5/20) , and there was significant differ-ence( P <0. 05). The incidence of buccal gingival sulcus becoming shallow in treatment group was 5. 5% (1/ 18), which was markedly higher than that in control group(90% , 18/20), and there was significant difference ( P <0. 01). [Conclusion]The efficacy of heterogeneous acellular dermal matrix(Oral Biofilm) for repairing hard palatal fistula is satisfactory, and has simple operation, little damage of surrounding tissue and no incidence of buccal gingival sulcus becoming shallow after the operation.%[目的]探讨异种脱细胞真皮基质(下称口腔修复膜)修补硬腭部腭瘘的临床疗效.[方法]38例硬腭部腭瘘的患者随机分为治疗组和对照组,治疗组18例患者采用口腔修复膜修补硬腭部腭瘘,对照组20例患者采用传统方法修复硬腭部腭瘘,观察两组患者术后1~3个月手术部位是否再次穿孔及龈颊沟是否变浅.[结果]治疗组在术后1~3个月再次穿孔的发生率为11.1%(2/18)显著低于对照组25.0%(5/20),其差异有统计学意义(P<0.05);治疗组龈颊沟变浅发生率为5.5%(1/18)显著低于对照组发生率为90%(18

  18. 微孔结构对猪脱细胞真皮基质血管化影响的初步观察%Effect of laser micropore structure on vascularization of porcine acellular dermal matrix

    Institute of Scientific and Technical Information of China (English)

    曾逃方; 罗旭; 林才; 何勇; 曾元临; 辛国华

    2012-01-01

    Objective To observe the effect of laser micropore structure on vascularization of porcine acellular dermal matrix.Methods 36 healthy male nude mice were randomly divided into experimental group and control group.The experimental group were transplanted laser micropore porcine acellular dermal matrix (LPADM) and autologous thin skin,the control group were transplanted non-porous acellular dermal matrix and autologous thin skin,the surgery were completed by two-step.Each group select six nude mice to cut specimen for histology and electron microscopy at 1 d,3 d,14 d after operation.Results Scanning electron microscopy revealed new tissue ingrowth into LPADM by laser micropore structure of the experimental group at 1 d after operation,histological observation showed that there were vessel-like lumen structure in the new organization,and the new organization was obvious ingrowth at 3 d after operation.At 14 d after operation,the LPADM completed revascularization,the new organization in the laser microporous gradually changed into the collagen fiber-based dermal tissue.During the whole experiment,the was no blood vessels or vascular endothelial cells move in the control group.Conclusions The presence of laser microporous structure can improve the vascularization capacity of LPADM,provide a channel of the cells and organization ingrowth.%目的 初步观察激光微孔结构对猪脱细胞真皮基质(ADM)血管化的影响.方法 健康雄性裸鼠36只,随机分为实验组及对照组,实验组移植激光微孔猪脱细胞真皮基质(LPADM)加自体薄皮片,对照组移植无孔猪脱细胞真皮基质加自体薄皮片,手术分“二步法”完成.术后1d、3d、14 d每组各取6只裸鼠处死,切取标本行组织学及电镜检查.结果 实验组术后第1天,扫描电镜发现新生组织通过微孔结构长入ADM内部.组织学观察显示新生组织中有血管样管腔结构,其内有红细胞征象.术后第3天实验组LPADM中新生组织明显

  19. Advantages of implantation of acellular porcine-derived mesh in the treatment of human rectocele – Case report

    Directory of Open Access Journals (Sweden)

    Tomasz Kościński

    2016-09-01

    Full Text Available Introduction. A rectocele is a hernation of the rectum into the vaginal lumen developing as a consequence of weakness of the rectovaginal septum. It affects about 18% of women after childbearing age. Symptoms associated with a rectocele include constipation, vaginal fullness or heaviness, feeling of a bulging mass within vagina, incomplete stool evacuation and dyspareunia. Current methods of surgical treatment of a rectocele often require implantation of a mesh graft. In most of cases, synthetic and non-absorbable meshes are used. Although implantation of a synthetic and non-absorbable mesh is effective in the treatment of rectocele, a high rate of mesh erosion has been reported. Case report. This study presents a surgical technique and case report for the treatment of a rectocele in a 46-year-old women by implantation of a porcine-derived absorbable collagen mesh (Pelvicol® by transvaginal approach, with six year follow-up. A review of the literature concerning implantation of Pelvicol® for the treatment of rectocele was also undertaken. Conclusions. The clinical experience and review of the literature by the authors suggest that a porcine-derived acellular mesh is non-cytotoxic, pyrogenic or allergenic, and the application of a biomesh in the management of rectocele is effective and safe, and the risk of mesh erosion is very low.

  20. The application of acellular dermal matrix combined with coralline hydroxyapatite in guided bone regeneration%异种脱细胞真皮基质联合珊瑚羟基磷灰石在GBR术中的应用

    Institute of Scientific and Technical Information of China (English)

    汪竹红; 康博; 黄达鸿; 管红雨; 温玉洁; 林天赐; 林丽娥

    2014-01-01

    Objective: To evaluate the clinical effectiveness of acellular dermal matrix combined with coralline hydroxyapatite for guided bone regeneration. Methods: 17 patients with 27 lost teeth were included in this study. 10 anterior lost teeth area with the alveolar bone thickness about 4mm was placed ankylos implants using bone condensing technique followed by guided bone generation. The other lost teeth area with bone defect was placed ankylos implants using routine method followed by GBR. 6-8 months later, the second-stage operation was performed and the condition of the new bone was observed. Results: All implants showed good osseointegration and were covered by alveolar bone except one implant whose labial neck about 1.5mm height wasn't covered by bone. Conclusion:Acellular dermal matrix combined with coralline hydroxyapatite can achieve good bone formation in guided bone regeneration.%目的:评价异种脱细胞真皮基质联合珊瑚羟基磷灰石在引导骨组织再生术中的应用效果。方法:17例共27颗牙缺失患者作为研究对象,其中10颗上前牙牙槽骨宽度约4mm的延期种植先行骨挤压术植入种植体再行GBR术,其余12颗延期即刻种植上前牙及5颗环状骨缺损后牙常规植入种植体后行GBR术。6-8m后观察成骨效果。结果:除一例患者右上侧切牙植体颈部唇侧暴露约1.5mm左右,其余患者植体均被新生骨包绕,成骨效果显著。结论:异种脱细胞真皮基质联合珊瑚羟基磷灰石在牙种植术中引导骨组织再生效果良好。

  1. Acellular dermis-assisted prosthetic breast reconstruction: a systematic and critical review of efficacy and associated morbidity.

    Science.gov (United States)

    Sbitany, Hani; Serletti, Joseph M

    2011-12-01

    The use of acellular dermal matrix to assist in two-stage expander/implant breast reconstruction has increased over recent years. However, there are questions regarding the potential for increased morbidity when using these techniques relative to standard submuscular coverage techniques. This systematic review combines published data comparing the techniques, to compare morbidity and advantages of acellular dermal matrix relative to standard submuscular coverage techniques. An English language literature search was performed to find articles reporting outcomes of two-stage expander/implant reconstruction using acellular dermal matrix. The outcome categories analyzed were patient/treatment demographics, tissue expander characteristics, and complications. Nine articles met inclusion criteria for this analysis. Six of these were matched cohort studies comparing outcomes of acellular dermal matrix techniques to standard submuscular techniques. The remaining three were case series of acellular dermal matrix techniques. The only difference found in complications was a higher rate of seroma for the acellular dermal matrix group (4.3 percent versus 8.4 percent, p = 0.03). Despite this, both groups illustrated similar rates of infection leading to explantation (3.2 percent for submuscular and 3.4 percent for acellular dermal matrix, p = 0.18). In addition, acellular dermal matrix techniques illustrated greater intraoperative fill volumes and consistently fewer fills required to reach expander capacity. The use of acellular dermal matrix in two-stage expander/implant reconstruction offers a safety profile similar to that of standard submuscular techniques. Both techniques have shown similar rates of infection ultimately requiring explantation. In addition, acellular dermal matrix offers the advantage of a more rapid reconstruction with less need for manipulation of the prosthetic through filling. Therapeutic, III.

  2. Hair Follicle Morphogenesis in the Treatment of Mouse Full-Thickness Skin Defects Using Composite Human Acellular Amniotic Membrane and Adipose Derived Mesenchymal Stem Cells

    Directory of Open Access Journals (Sweden)

    Wu Minjuan

    2016-01-01

    Full Text Available Early repair of skin injury and maximal restoration of the function and appearance have become important targets of clinical treatment. In the present study, we observed the healing process of skin defects in nude mice and structural characteristics of the new skin after transplantation of isolated and cultured adipose derived mesenchymal stem cells (ADMSCs onto the human acellular amniotic membrane (AAM. The result showed that ADMSCs were closely attached to the surface of AAM and grew well 24 h after seeding. Comparison of the wound healing rate at days 7, 14, and 28 after transplantation showed that ADMSCs seeded on AAM facilitated the healing of full-thickness skin wounds more effectively as compared with either hAM or AAM alone, indicating that ADMSCs participated in skin regeneration. More importantly, we noticed a phenomenon of hair follicle development during the process of skin repair. Composite ADMSCs and AAM not only promoted the healing of the mouse full-thickness defects but also facilitated generation of the appendages of the affected skin, thus promoting restoration of the skin function. Our results provide a new possible therapy idea for the treatment of skin wounds with respect to both anatomical regeneration and functional restoration.

  3. Hair Follicle Morphogenesis in the Treatment of Mouse Full-Thickness Skin Defects Using Composite Human Acellular Amniotic Membrane and Adipose Derived Mesenchymal Stem Cells

    Science.gov (United States)

    Minjuan, Wu; Jun, Xiong; Shiyun, Shao; Sha, Xu; Haitao, Ni

    2016-01-01

    Early repair of skin injury and maximal restoration of the function and appearance have become important targets of clinical treatment. In the present study, we observed the healing process of skin defects in nude mice and structural characteristics of the new skin after transplantation of isolated and cultured adipose derived mesenchymal stem cells (ADMSCs) onto the human acellular amniotic membrane (AAM). The result showed that ADMSCs were closely attached to the surface of AAM and grew well 24 h after seeding. Comparison of the wound healing rate at days 7, 14, and 28 after transplantation showed that ADMSCs seeded on AAM facilitated the healing of full-thickness skin wounds more effectively as compared with either hAM or AAM alone, indicating that ADMSCs participated in skin regeneration. More importantly, we noticed a phenomenon of hair follicle development during the process of skin repair. Composite ADMSCs and AAM not only promoted the healing of the mouse full-thickness defects but also facilitated generation of the appendages of the affected skin, thus promoting restoration of the skin function. Our results provide a new possible therapy idea for the treatment of skin wounds with respect to both anatomical regeneration and functional restoration. PMID:27597871

  4. In vitro investigation on morphological and physico-chemical properties of particulate acellular dermal matrix%颗粒状脱细胞真皮基质的形态学特征及理化性能体外研究

    Institute of Scientific and Technical Information of China (English)

    左海斌; 彭代智; 郑必祥; 何升东; 周新; 刘敬

    2011-01-01

    目的 研究颗粒状脱细胞真皮基质(particulate acellular dermal matrix,PADM)的体外形态学特征与理化性能.方法 收集SD大鼠背部皮肤样本,采用本实验室的脱细胞方法制备片状脱细胞真皮基质(acellular dermal matrix,ADM)并分析其内胞核和DNA残留情况.将片状ADM切割成不同规格的PADM,并结合大体观察、组织学分析、电镜技术、激光粒度分析仪和傅里叶变换红外光谱仪检测其外观轮廓、形态学、粒径分布和胶原分子结构.并将其复合人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)体外培养24 h,观察PADM与HUVEC的黏附情况.结果脱细胞方法有效降低了ADM的DNA含量[(1.51±0.37) μg/mg vs (0.30±0.09) μg/mg].制备的PADM为白色颗粒,外形近似于长方体或立方体,显微镜下HE染色观察显示颗粒内部胶原纤维束结构疏松.扫描电镜显示颗粒断面结构疏松,透射电镜显示胶原原纤维周期性横纹结构和均匀分布的胶原原纤维间隙.激光粒度分析仪检测显示其粒度分布集中,例如,规格为0.2 mm 的PADM,其80%(d0.1 ~ d0.9)的粒径分布于233~487 μm.FTIR结果显示,PADM分别在1 659、1 549、1 239 cm-1处出现的特征吸收峰,表明其保留了胶原分子的α-helix、β-sheet和β-turn二级结构,且体外HUVEC较易黏附于PADM.结论室温条件下,切割法制备的新型PADM形态规则,粒径分布集中,胶原纤维束形态和胶原分子的二级结构保存良好.%Objective To investigate the morphological and physico-chemical properties of particulate acellular dermal matrix (PADM) in vitro. Methods Dorsal skin samples were collected from SD rats, and then were deeellularized to obtain acellular dermal matrix (ADM) sheets. The ADM sheets were cut into PADM in different sizes. Digital camera, light microscope (HE staining) and electron microscopes were used to observe the morphological properties of PADM. Laser diffraction and Fourier

  5. 缩水甘油改性pADM的亲水性与吸湿动力学研究%Hydrophility and Kenetic Analysis of Moisture Adsorption of Porcine Acellular Dermal Matrix Modified by Glycidol

    Institute of Scientific and Technical Information of China (English)

    但年华; 肖世维; 但卫华; 林海; 朱剑

    2011-01-01

    采用缩水甘油(Glycido1)对脱细胞猪真皮基质(pADM)进行改性,通过对改性前后材料的接触角和吸湿动力学的研究,探寻改性前后材料亲水性能的变化.研究结果表明:pADM经缩水甘油改性后,材料的接触角降低.材料的吸湿动力学特征符合二级吸附动力学方程,表明吸湿过程属于多分子层吸附;且平衡吸附量随着改性剂用量的增加而提高.两者均表明,改性后材料的亲水性能增加.从而论证了通过引入亲水基团,提高pADM亲水性能的可行性.为进一步制备出多功能pADM,奠定了实验基础.%The contact angle and moisture adsorption of the porcine acellular dermal matrix (pADM) were mensurated after modified by Glycidol.The improvement of surface hydrophilicity was proved by the declined of contact angle in the surface of pADM.And the adsorption process could be characterized by the Second-Order Model of adsorption kinetic equation after kinetic analysis of moisture adsorption under RH85% and RH43%.The results inducated that the adsorption was belong to multilayer adsorption.The decreased contact angle and increased equilibrium adsorption capacity of water implied that the hydrophilicity of pADM could be developed by introducing more polar groups into pADM.

  6. 脱细胞异体真皮基质填塞治疗肛瘘的临床体会%Clinical study on anal fistula treatment with acellular extracellular matrix

    Institute of Scientific and Technical Information of China (English)

    安少雄; 黄斌

    2012-01-01

    Objective To observe the effects of the efficacy of acellular extracellular matrix (AEM) treatment for anal fistula. Methods Twelve patients with anal fistula were treated with AEM. The clinical effects were observed. Results Nine cases were cured, but 3 failed and 1 case relapsed after 6 months. The heal rate of anal fistula was 75%. Conclusions This method has advantages of mild traumas and less pain, rapid recovery rapidly, and the most important is that this treatment could protect anal function and appearance. However, the long-term effect needs further study.%目的 探讨脱细胞异体真皮基质填塞治疗肛瘘的临床疗效.方法 采用脱细胞异体真皮基质填塞治疗12例肛瘘患者,观察其临床效果.结果 本组患者瘘管愈合时间7~18 d,平均(13.0±3.3)d.12例患者中治愈9例,治愈率为75%,失败3例,随访6个月复发1例,复发率为8.3%.本组无肛门失禁、肛门畸形发生.结论 脱细胞异体真皮基质填塞治疗肛瘘具有创伤小、疼痛轻、恢复快、不损害肛门功能和外形的优点,但远期效果需进一步观察.

  7. Comparative evaluation of the effectiveness of acellular dermal matrix allograft and subepithelial connective tissue to coronally advanced flap alone in the treatment of multiple gingival recessions: A clinical study

    Directory of Open Access Journals (Sweden)

    Pallavi Thakare

    2015-01-01

    Full Text Available Background: Obtaining predictable and esthetic root coverage has become an important part of periodontal therapy. Several techniques have been developed to achieve these goals with variable outcomes. The aim of this study was to appraise the effectiveness of acellular dermal matrix allograft (ADMA and subepithelial connective tissue graft (SCTG compared to coronally advanced flap (CAF in the treatment of multiple gingival recessions. Materials and Methods: A total of 30 patients aged between 18 and 50 years, with multiple Miller's Class I and II recessions on labial or buccal surfaces of teeth were selected for this study. The patients were randomly assigned to CAF + ADMA, CAF + SCTG and CAF groups with 10 patients in each group. The clinical parameters assessed were probing pocket depth (PPD, clinical attachment level (CAL, gingival recession (GR, width of keratinized tissue, plaque index and papilla bleeding index at base line and 6 months after surgery. Results: Statistical analysis using One-way ANOVA suggested that the root coverage obtained was greater in the ADMA + CAF (89.83 ± 15.29%, when compared to SCTG + CAF (87.73 ± 17.63% and CAF (63.77 ± 27.12% groups. The predictability for coverage of> 90% was greater in CAF + ADMA (65% when compared with SCTG + CAF (61.66% and CAF (31.17%. Improvements in the clinical parameters from baseline were found in all the three groups treated. Conclusion: It was concluded that all three techniques could provide root coverage in Miller's class I and II gingival recessions; but greater % root coverage and predictability for coverage of> 90% could be expected with CAF + ADMA and CAF + SCTG groups when compared with CAF alone.

  8. 应用脱细胞真皮基质材料修补复杂腹壁切口疝%Repair of complex abdominal incisional hernia with acellular dermal matrix

    Institute of Scientific and Technical Information of China (English)

    李小军; 王小强; 龙延滨; 邱健; 张瑞鹏

    2011-01-01

    目的 探讨脱细胞真皮基质材料对复杂的腹壁切口疝的修复治疗效果.方法 回顾性分析2008年1月至2010年6月间使用脱细胞真皮基质(acellular dermal matrix,ADM)材料修补的7例复杂的腹壁切口疝的治疗方法.其中男4例,女3例,年龄43~83岁,中位年龄53岁;7例中有2例伴有腹股沟斜疝,给予同时修补;5例同时进行了胃肠道手术,其中有2例伴有小肠瘘;疝环直径为9.2 ~16.5 cm,平均(11.6±2.8)cm;5例使用腹腔内修补(intraperitoneal onlay mesh,IPOM),2例为腹膜外修补(total extraperitoneal prothesis,TEP).结果 本组患者均手术顺利,放置ADM补片至关腹结束的平均手术时间(33±12) min;术中平均出血量(16±4) ml;住院时间7~12d.所有使用ADM的患者均痊愈出院,术后未发现有慢性疼痛、感觉异常、肺炎、尿路感染等并发症,手术切口无红肿、溃破、无血清肿.7例均获随访,随访时间5 ~26个月,中位随访时间为14个月,随访期间未发现浅部感染或深部感染,无疝复发.结论 脱细胞真皮基质材料作为一种新的生物补片,适用于复杂腹壁切口疝,尤其是伴有污染的腹壁切口疝的修补.%Objective To evaluate the repair of abdominal complicated incisional hernia using acellular dermal matrix (ADM).Methods Retrospective analysis was made on 7 cases with abdominal complicated incisional hernia treated by ADM in our hospital from January 2008 to June 2010,among them there were 4 males and 3 females.Age ranged from 43 to 83 years and the median age was 53 years.Two concurrent indirect inguinal hernia cases were repaired and concurrent gastrointestinal tract problems including 2 small bowd fistulas were operated one stage in 5 cases.Mean diameter of hernia ring was ( 11.6 ± 2.8 ) cm,ranged from 9.2 to 16.5 cm.5 cases were repaired by using intraperitoneal onlay mesh,others using total extraperitoneal prothesis.Results All patients were operated on successfully

  9. 从脱细胞猪真皮基质材料的边角料中提取Ⅰ型胶原%Collagen Extracted from Offcut of Porcine Acellular Dermal Matrix

    Institute of Scientific and Technical Information of China (English)

    刘新华; 路翠娟; 但年华; 刘婷; 胡杨; 但卫华

    2013-01-01

    以脱细胞猪真皮基质材料的边角料为原料,使用酸-酶结合法提取Ⅰ型胶原,并用SDS-PAGE电泳、DSC、FI-IR、SEM、AFM对Ⅰ型胶原进行了结构表征.研究表明:组织学观察发现,脱细胞猪真皮基质材料中不存在细胞和细胞碎片,且胶原纤维得到充分的分离和松散,非纤维成分大部分被水解,有利于Ⅰ型胶原的提取.提取得到的Ⅰ型胶原红外特征峰明显,分子质量大且分布窄,变性温度为69.4℃;SEM显示胶原呈现孔隙均匀的三维网状结构;AFM观察到胶原大部分排列紧密,相互编织缠绕,形成无规线团,小部分胶原直径较粗,且以线性分布,说明胶原已经发生自聚集现象.综合检测分析结果,可以认为所提取制备得到的确为具有天然三股螺旋结构的Ⅰ型胶原.%The histological changes of fresh pigskin after acellular were observed and analyzed by hematoxylin-eosin staining (HE) and iron hematoxylin staining.Using the offcut of porcine acellular dermal matrix as a raw material,collagen was extracted with the method of acid swelling-pepsin digestion.Then the obtained collagen was characterized by SDS-PAGE,FT-IR spectra,Differential Scanning Calorimeter (DSC),Atomic Force Microscope (AFM) and Scanning Electron Microscopic (SEM).The study shows that:the cells in fresh pigskin are removed completely after the treatment process,the collagen fibers are loosed well,and most of the non-fiber component is hydrolyzed,which is conducive to the extraction of collagen.The extracted type Ⅰ collagen has its characteristic peaks of FI-IR,and the molecular weight is of not only large,but also narrow distribution.The denaturation temperature is 69.4℃ ; the SEM displayed that the collagen remains three-dimensional mesh structure with uniform pore.We could also conclude that the collagen has closely arranged,mutually braided or wound to form a random coil.The diameter of the small portion of the collagen is coarse,and is in a

  10. 异种脱细胞真皮基质联合Bio-oss Collagen修复牙槽骨缺损的临床研究%Clinical application of acellular dermal matrix combined with Bio-oss Collagen to repair alveolar bone defects

    Institute of Scientific and Technical Information of China (English)

    杨春羚; 林良缘; 庄亮亮; 曾金表

    2011-01-01

    Objective: To investigate the effect of guided bone regeneration of acellular dermal matrix (ADM) combined with Bio -oss Collagen in alveolar defect. Method: 18 cases patients with severe alveolar bone resorption or damage were included.Bone defect after teeth extraction were very serious and the residual height and width of the alveolar bone were very limited in all these cases. Bio-oss Collagen was delivered into the sockets immediately after tooth extraction and covered with acellular dermal matrix membrane.Suture was removed 2 weeks postop.and the alveolar bone was regularly examined at the 3rd month. Result:New bone was found to form well in both physical examination and X-ray examination in all the 18 cases in the 3rd month.The height and width of the alveolar bone were significantly increased, providing a good bone condition for later FPD prosthetic treatment. Conclusion: Acellular dermal matrix combined with Bio-oss Collagen can improve the bone condition before restoration in clinical.%目的:通过引导骨再生(GBR)技术评估异种脱细胞真皮基质(acellular dermal matrix,ADM)与Bio-oss Collagen联合应用在修复牙槽骨缺损中的作用.方法:选择拔牙术后牙槽骨缺损严重的病例18例,拔牙同期在拔牙创植入Bio-oss Collagen并覆盖异种脱细胞真皮基质(海奥生物膜),术后2周拆线,3个月复诊并拍摄X线片.结果:术后经临床检查和X线检查,18例患者植骨区新骨形成良好,牙槽骨高度与丰满度明显改善,术区骨生成良好.结论:临床上异种脱细胞真皮基质与Bio-oss Collagen联合应用能有效修复牙槽骨缺损,改善修复前的骨条件.

  11. The curative effect of acellular allodermis matrix on scar proliferation of patients with deep burn wounds of function positions%脱细胞异体真皮对功能部位深度烧伤创面愈合瘢痕增生的改善作用

    Institute of Scientific and Technical Information of China (English)

    蒙诚跃; 王润秀; 梁自乾; 张立明; 汪永连

    2003-01-01

    @@ BACKGROUND:Because there was no thick skin for skin grafting supplied by enough area of supplying skin of patients with extensive area burn only autograft skin particle,split thickness autologous skin graft or cultural autograft can be applied in repairing wound,which cannot resolve the problems of contracture and deformity caused by scar proliferation after healing of wounds and dysfunction of joint.Acellular allodermis matrix of J 1 type is a kind of tissue with lowest antigenicity and thought as an ideal substituted material for resolving difficult problem of scar proliferation and dysfunction.

  12. Allogenic acellular extracellular matrix repairs high anal fistula%脱细胞异体真皮基质填塞修复高位肛瘘

    Institute of Scientific and Technical Information of China (English)

    王健诚; 王炜; 张科; 邹世镇

    2014-01-01

    背景:治疗高位肛瘘的方法有瘘管剔除、切开挂线、选择性黏膜瓣推移、生物蛋白胶封堵、括约肌间瘘管结扎等,大都存在创面愈合间长、一次成功率较低、复发率偏高的不足,术后并发症发生率高。  目的:观察应用脱细胞异体真皮基质治疗高位肛瘘的临床疗效,探讨治疗高位肛瘘的微创治疗新方法。  方法:选择100例高位肛瘘患者,根据患者意愿分2组治疗,治疗组采用脱细胞异体真皮基质填塞治疗,对照组采用传统的肛瘘低位切开并高位挂线治疗,比较两组手术时间、术中出血量、术后目测类比评分、术后疼痛持续时间、肛门失禁严重程度评分、创面愈合时间、一期手术成功率、治愈率及复发率。  结果与结论:治疗组手术时间、术中出血量、创面愈合时间、术后目测类比评分、术后疼痛持续时间、肛门失禁严重程度评分均低于对照组(P OBJECTIVE:To observe the clinical effect of acelular extracelular matrix in the treatment of high anal fistula, and to explore a minimaly invasive treatment for high anal fistula. METHODS: Totaly 100 cases of high anal fistula were randomly divided into treatment group and control group, 50 cases in each group. Treatment group were treated with alogenic acelular extracelular matrix, and control group were treated with traditional low incision with high thread-drawing. Then, we observed and compared the operation time, bleeding volume, postoperative pain score (visual analog scale score), postoperative pain duration, anal incontinence severity score (Wexner score), wound healing time, one-stage success rate, cure rate, recurrence rate. RESULTS AND CONCLUSION: Compared with the control group, the treatment group showed lower scores in the operation time, bleeding volume, wound healing time, visual analog scale score, postoperative pain duration, and anal incontinence severity score

  13. Two-stage implant-based breast reconstruction compared with immediate one-stage implant-based breast reconstruction augmented with an acellular dermal matrix : An open-label, phase 4, multicentre, randomised, controlled trial

    NARCIS (Netherlands)

    Dikmans, Rieky E. G.; Negenborn, Vera L.; Bouman, Mark-Bram; Winters, Hay A. H.; Twisk, Jos W. R.; Ruhe, P. Quinten; Mureau, Marc A M; Smit, J.M.; Tuinder, Stefania; Eltahir, Yassir; Posch, Nicole A.; van Steveninck-Barends, Josephina M.; Meesters-Caberg, Marleen A.; van der Hulst, Rene R. W. J.; Ritt, Marco J. P. F.; Mullender, Margriet G.

    Background The evidence justifying the use of acellular dermal matrices (ADMs) in implant-based breast reconstruction (IBBR) is limited. We did a prospective randomised trial to compare the safety of IBBR with an ADM immediately after mastectomy with that of two-stage IBBR. Methods We did an

  14. 导入透明质酸猪脱细胞真皮基质的刺激性及致敏性研究%Skin irritation and sensitization of swine acellular dermal matrix treated with hyaluronic acid

    Institute of Scientific and Technical Information of China (English)

    宁少南; 赵筱卓; 王慧英; 张国安

    2012-01-01

    Objective To evaluate the skin irritation and sensitization potential of the swine acellular dermal matrix treated with hyaluronic acid (SADM-HA).Methods (1) Skin irritation test.Twelve New Zealand rabbits were divided into SADM-HA group,allogeneic skin group,and (human) xeno-skin group according to the random number table,with 4 rabbits in each group.Four test sites were designed on the back of each rabbit.Two test sites of each rabbit in the three groups were covered with SADM-HA,allogeneic skin,and xeno-skin,respectively.Another test site was covered with gauze containing 200 g/L sodium dodecyl sulfate solution as positive control.The last test site was covered with gauze containing normal saline as negative control.The primary irritation index and cumulative irritation index of each material were calculated.(2) Skin closed-patch test.Sixty guinea pigs were used.Fifty-four guinea pigs were divided into SADM-HA group,allogeneic skin group,and (human) xeno-skin group according to the random number table,with 18 guinea pigs in each group.Twelve guinea pigs in each of the three groups were correspondingly induced and stimulated by SADM-HA,allogeneic skin,and xeno-skin,with 6 guinea pigs in each group treated with ethanol-soaked gauze to serve as negative control.The remaining 6 guinea pigs were treated with gauze containing 25% α-hexylcinnamaldehyde ethanol solution as positive control.The rating scales of Magnusson and Kligman were used to grade the condition of skin after being treated with above-mentioned materials to evaluate skin sensitivity to them at post stimulation hour 24 and 48.Data were processed with the nonparametric test of independent samples.Results (1) In the skin irritation test,the primary irritation indexes of the three dressings in SADM-HA group,allogeneic skin group,and xeno-skin group were respectively-0.04,0.13,and 0.08.The cumulative irritation indexes of the three dressings in SADM-HA group,allogeneic skin group,and xeno-skin group were

  15. Regeneration of human epidermis on acellular dermis is impeded by small-molecule inhibitors of EGF receptor tyrosine kinase.

    Science.gov (United States)

    Forsberg, Sofi; Ostman, Arne; Rollman, Ola

    2008-10-01

    The family of human epidermal growth factor receptors (EGFR, HER2-4) exerts key functions in normal and malignant epithelial cells. Both EGFR and HER2 are valuable targets for anti-cancer drugs by interfering with ligand binding, receptor dimerization, or tyrosine kinase activity. A similar therapeutic strategy has been advocated for chronic psoriasis since plaque lesions overexpress EGFR and its ligands. Our aim was to characterize EGFR/HER2 protein expression in skin cultures and to evaluate the effects of tyrosine kinase inhibitors on epidermal outgrowth, morphology, and EGFR activation. Human skin explants were established on cell-free dermis and cultured at the air-liquid interface. The impact of small-molecule HER inhibitors on outgrowth was assayed by fluorescence-based image analysis and histometry. Effects of a dual EGFR/HER2 kinase inhibitor, PKI166, on neoepidermis were studied by immunohistochemistry and Western blot. Receptor immunostaining showed in vivo-like distributions with highest EGFR intensity in the proliferative layers whereas HER2 was mainly expressed by suprabasal keratinocytes. Reepithelialization was associated with EGFR autophosphorylation irrespective of exogenous ligand stimulation. PKI166 inhibited neoepidermal EGFR activation, keratinocyte proliferation, and outgrowth from normal and psoriatic skin explants. The rate of epidermalization in presence of other HER inhibitors varied suggesting that drug specificity, potency, and reversibility determine the dynamic outcome. Overall, agents predominantly targeting EGFR kinase were more efficient inhibitors of epidermal regeneration than an HER2-selective drug. The study illustrates the usefulness of a dynamic skin model and emphasizes the potential of HER-directed approaches to control epidermal growth in hyperproliferative skin disorders.

  16. Engineering an improved acellular nerve graft via optimized chemical processing.

    Science.gov (United States)

    Hudson, Terry W; Liu, Stephen Y; Schmidt, Christine E

    2004-01-01

    The long-term goal of our research is to engineer an acellular nerve graft for clinical nerve repair and for use as a model system with which to study nerve-extracellular matrix interactions during nerve regeneration. To develop this model acellular nerve graft we (1) examined the effects of detergents on peripheral nerve tissue, and (2) used that knowledge to create a nerve graft devoid of cells with a well-preserved extracellular matrix. Using histochemistry and Western analysis, the impact of each detergent on cellular and extracellular tissue components was determined. An optimized protocol was created with the detergents Triton X-200, sulfobetaine-16, and sulfobetaine-10. This study represents the most comprehensive examination to date of the effects of detergents on peripheral nerve tissue morphology and protein composition. Also presented is an improved chemical decellularization protocol that preserves the internal structure of native nerve more than the predominant current protocol.

  17. 新型激光微孔化猪脱细胞真皮基质的制备及应用%Preparation and application of novel laser micropore porcine acellular dermal matrix

    Institute of Scientific and Technical Information of China (English)

    罗旭; 万丽; 李安乐; 徐建军; 夏卫东; 李玉莉; 邓春林; 王平; 林才

    2012-01-01

    目的 介绍一种新型微孔化猪脱细胞真皮基质并验证其安全性、可行性.方法 取新鲜健康小白猪断层真皮,采用高渗盐-胰蛋白酶法脱细胞,经超声波充分震荡洗涤,获得猪脱细胞真皮基质,经特定激光微孔技术,在脱细胞真皮基质上贯穿打孔即获得新型激光微孔化猪脱细胞真皮基质.将24只SD大鼠均切除背部全层皮肤至深筋膜,造成大小2.0 cm×2.0 cm创面,随机分为实验组和对照组,每组各12只,同步I期行自体中厚皮片与真皮材料复合移植于大鼠全层皮肤缺失创面,实验组采用移植真皮材料为激光微孔化猪脱细胞真皮基质(LPADM),对照组采用移植真皮材料为普通的猪皮脱细胞真皮基质(ADM),组织学检查、电镜观察两组真皮材料的物理性状、结构完整性、细胞残留及移植后皮片活性、血管化和移植后动物安全性变化.结果 制备的真皮基质材料LPADM和ADM均呈瓷白色,有光泽,柔软而有弹性;组织学检查未见上皮细胞、内皮细胞残留,结构完整性好;电镜检查胶原纤维排列整齐,保持较好的结构完整性.实验组术后7 d LPADM组织学检查显示移植真皮血管化充分,术后14 d复合移植皮片均存活;对照组术后7 d移植皮片色暗淡,多有起泡,并逐渐失活坏死,ADM组织学检查未见新生血管形成,术后14 d复合移植皮片干性坏死.两组实验动物均无异常死亡或感染,亦未见皮肤过敏反应,体重无减轻.结论 以特定激光微孔化等技术制备的新型猪脱细胞真皮基质动物移植实验中血管化充分、复合皮片愈合良好,获得满意的实验效果.%Objective To prepare a new type of micropore porcine acellular dermal matrix with the aid of laser (PLADM ), and validate the safety and practicability of the PLADM. Methods LPADM was prepared by a specific laser micropore technology with the punch worked in cycles and suspended in the midair on the harvested split

  18. Human Homeostatic Control Matrix in Norm

    Directory of Open Access Journals (Sweden)

    Alexander G. Kruglov

    2016-09-01

    Full Text Available We undertook our research to study and systemize the relationship between hemodynamics and biochemical parameters of arterial and venous blood in healthy people. Hemodynamic and biochemical characteristics were obtained through a probe by using catheterization in various vascular areas (aorta, brain, heart, lungs, and liver. Correlation and factor analyses were conducted to study the relationship between the obtained characteristics of the regional and systemic blood flow. Due to the nature of the correlation analysis, the significant (p<0.05 relation signs (+, 0, - without regard to their power were considered. The obtained results suggested that there are sets of both intra-organ and system regulatory relationships between metabolic and hemodynamic characteristics. The complex of relationships among the studied parameters makes it possible to maintain the homeostatic equilibrium in the body. The psychophysiological control system includes the subsystems we described: 1 the cardiac-hepatic-pulmonary complex having properties of the metabolic and hemodynamic information field providing biological stability of the homeostasis; any significant imbalance of its elements triggers afferent information flows actualizing an afferent synthesis; 2 the mind forming gradient patterns of targeted behavior to eliminate metabolic imbalance, to achieve goals both as coded biological parameters and as the highest forms of behavior, to reach the ultimate goal: parametric, homeostatic equilibrium in the “biosphere” of the human body. By using the results of our research and the complex of dynamic relationships in human homeostasis, we built a homeostatic control matrix (HCM.

  19. Extra cellular matrix features in human meninges.

    Science.gov (United States)

    Montagnani, S; Castaldo, C; Di Meglio, F; Sciorio, S; Giordano-Lanza, G

    2000-01-01

    We collected human fetal and adult normal meninges to relate the age of the tissue with the presence of collagenous and non-collagenous components of Extra Cellular Matrix (ECM). Immunohistochemistry led us to observe some differences in the amount and in the distribution of these proteins between the two sets of specimens. In particular, laminin and tenascin seem to be expressed more intensely in fetal meninges when compared to adult ones. In order to investigate whether the morphofunctional characteristics of fetal meninges may be represented in pathological conditions we also studied meningeal specimens from human meningiomas. Our attention was particularly focused on the expression of those non-collagenous proteins involved in nervous cell migration and neuronal morphogenesis as laminin and tenascin, which were present in lesser amount in normal adult specimens. Microscopical evidences led us to hipothesize that these proteins which are synthesized in a good amount during the fetal development of meninges can be newly produced in tumors. On the contrary, the role of tenascin and laminin in adult meninges is probably only interesting for their biophysical characteristics.

  20. Correction of penile curvature by allogeneic acellular dermal matrix%同种异体脱细胞真皮补片移植矫正白膜型阴茎弯曲的临床价值

    Institute of Scientific and Technical Information of China (English)

    董玉林; 吴小蔚; 田龙

    2012-01-01

    Objective To evaluate the effect of lengthening the short-side albuginea of the bending penis by the allogeneic acellular dermal matrix ( Allo-ADM ) for the treatment of penile curvature.Methods From Jun 2007 to Jun 2010,18 patients with penile curvature due to malformation of the albuginea cavernous body were treated.The age of the patients ranged from 15 to 26 years (mean,20 years).Twelve patients were married.The curvature degree ranged from 30° to 80° (mean,55°).There were 17cases of single curvature and 1 case of complex curvature.The grafts ( Allo-ADM ) of different sizes were sutured to the albuginea at the curvatus side of the penis according to the extent of penile curvature through a circumcision incision.The extent of penile curvature and complications were evaluated postoperatively.Results Penile curvature was corrected in all 18 patients after the operation.No infection,hematoma and abnormal erection occurred postoperatively. No erectile dysfunction and penile re-curvature was observed during the follow-up period of 3 to 24 months. Conclusion Lengthening the short-side albuginea of the bending penis by Allo-ADM could be a safe and effective way to correct penile curvature.%目的 评价应用同种异体脱细胞真皮补片移植矫正白膜型阴茎弯曲的安全性与疗效.方法 2007年6月至2010年6月收治白膜型阴茎弯曲患者18例,年龄15~26岁,平均20岁.已婚12例.阴茎弯曲度30°~80°,平均55°;单侧弯曲17例,复杂弯曲1例.硬膜外麻醉或全麻,包皮环切切口入路,应用同种异体脱细胞真皮补片移植,延长曲侧海绵体白膜的术式治疗. 结果 18例阴茎弯曲均得到勃起直视下矫正,矫正后阴茎弯曲度0°~10°,平均4°.术后无感染、血肿、局部结节等并发症.18例随访3~24个月,无勃起功能障碍,未见弯曲复发、勃起硬结和形态畸形. 结论 同种异体脱细胞真皮补片矫正白膜型阴茎弯曲具有手术安全、疗效可靠、并发症少等优点.

  1. 异种脱细胞真皮基质在口腔颌面部创面修复中的应用%Application of heterogeneous acellular dermal matrix in repairing oral and maxillofacial defect

    Institute of Scientific and Technical Information of China (English)

    邵小钧; 庞恋苏; 袁仕廷; 解涓; 席庆

    2015-01-01

    目的 评价异种脱细胞真皮基质修复膜用于口腔颌面部创面修复中的临床效果.方法 收集2011 - 2014年解放军总医院海南分院口腔科59例因肿瘤、外伤、黏膜病变、瘢痕切除术等原因引起的口腔颌面部缺损,应用异种脱细胞真皮基质修复膜进行修复,缺损部位为颊、腭、舌、口底、腮腺、牙龈、前庭沟等,术后随访2周~ 6个月,并进行术后追踪随访及修复效果评估.结果 共修复口腔颌面部各类创面59例,一期愈合52例,成功率达88.14%.7例因创面较大,且受植区创面不平整、不规则,与修复膜之间存在死腔,以及过早的张闭口运动等因素,导致修复膜与创面部分脱落.结论 异种脱细胞真皮基质在口腔颌面部各类创面修复中起到了创面覆盖、引导组织再生和支架作用,修复效果满意,值得临床推广.%Objective To evaluate the clinical effect of heterogeneous acellular dermal matrix (H-ADM) in oral and maxillofacial defect repair. Methods Fifty-nine cases with oral and maxillofacial defects which were caused by tumor surgery, trauma, mucosal disease and scar resection were restored with H-ADM. The defects were located in buccal region, palate, tongue,floor mouth, parotid gland, gingiva, oral vestibular groove and so on. All patients were followed up for 2 weeks to 6 months postoperatively and the repair efficacy was assessed.Results Of the 59 cases undergoing transplantation, 52 cases were primary healing with the success rate of 88.14%, 7 cases failed due to large, tough and irregular wound, cavity existing between repair membrane and wound, early mouth movement and so on, which caused the falling off of partial repaired membrane.ConclusionH-ADM plays a role in wound coverage, guide tissue regeneration and biological scaffold during wound healing. The effect of repairing is satisfactory and it is worthy of clinical promotion.

  2. Evaluation of chemical cross-linking method of porcine acellular dermal matrix with oxidative chitosan oligosaccha-ride%氧化壳寡糖交联脱细胞猪真皮基质的研究

    Institute of Scientific and Technical Information of China (English)

    王磊; 但年华; 陈一宁; 但卫华

    2016-01-01

    目的:对氧化壳寡糖(OCOS)交联脱细胞真皮基质(pADM)后的性能进行评价。方法将一定质量OCOS溶于缓冲溶液中,将pADM浸没在该体系中,在特定温度下交联改性一段时间。考察反应温度、pH值、用量和反应时间对基质材料收缩温度的影响,通过红外光谱、原子力显微镜、孔隙率、热稳定性、耐酶降解性、细胞毒性等对交联前后基质材料的结构、性能进行表征。结果最优交联改性条件为反应温度37℃,反应时间16 h,OCOS用量8%,pH 8.4。最优条件下交联改性,得到的材料(OCOS-pADM)收缩温度可以达到78.4℃,红外光谱中胶原的3个特征吸收峰仍然存在,原子力显微镜下可明显观察到纵向上的D周期明暗条纹,改性后材料孔隙率变大,差示扫描量热法表征改性后材料热变性温度达80.44℃,7 d后降解率仅为7.5%±1.7%,细胞毒性评级为1级。结论改性后基质各方面性能均有所提高,胶原天然结构没有遭到破坏,细胞毒性测试中细胞形态良好,初步具备作为生物材料所需的条件。%Objective To assess the properties of porcine acellular dermal matrix(pADM) before and affer cross-linked by ox-idative chitosan oligosaccharide (OCOS). Methods A certain quatity OCOS was dissolved into buffered solution, and pADM was soaked at certain temperature for a period of time. The effect of reaction temperature, pH, OCOS dosage and reaction time on shrinkage temperature(Ts) of matrix were observed. The structure and properties of the matrix material before and after cross-linking were evaluated by infrared spectroscopy, atomic force microscopy(AFM), porosity, thermal stability, collagenase degrada-tion and cytotoxicity. Results The best reaction condition of reaction temperature was 37 ℃, reaction time was 16-hour, O-COS dosage was 8%and pH was 8.4. Under the best reaction condition, Ts of OCOS-pADM was 78.4

  3. 双氧水-氢氧化钠结合对脱细胞猪真皮基质材料高度纯化的作用%Effects of H2 O2-NaOH on highly-purification of porcine-derived acellular dermal matrix

    Institute of Scientific and Technical Information of China (English)

    肖世维; 但年华; 但卫华

    2015-01-01

    Using traceable pigskin as raw material,by observing the change of thickness,area,collagen content, non-collagen content in effluent and the histological morphology,the optimum alkali swelling scheme,H 2 O 2 4%-NaOH 6%,of preparing acellular dermal matrix has been harvested.After the preparation of highly-puri-fied porcine-derived acellular dermal matrix,a series of tests have been carried out.The tensile strength of the material was 7.2 MPa.It has good water absorption ability and no heavy metal.Animal experiments demonstra-ted no obvious acute peroral toxicity and the cytotoxicity was only 1 grade.These results lay the foundation for the application of the highly-purified porcine-derived acellular dermal matrix in biomedical materials area in the future with further research.%以可溯源性猪皮为原料,采用特殊的化学、物理及生化方法对猪皮进行高度纯化处理,通过观察实验中皮块厚度、面积、废液中胶原蛋白与非胶原蛋白含量及组织学形貌变化,得出脱细胞猪真皮基质碱膨胀的最佳方案为双氧水4%-氢氧化钠6%.并制备了高纯度脱细胞猪真皮基质,检测其抗张强度为7.2 MPa,吸湿性能好,不含重金属,无经口急毒,细胞除去干净且细胞毒性1级,为今后脱细胞猪真皮基质在生物医用材料领域的应用奠定基础.

  4. The Human Pancreas as a Source of Protolerogenic Extracellular Matrix Scaffold for a New-generation Bioartificial Endocrine Pancreas.

    Science.gov (United States)

    Peloso, Andrea; Urbani, Luca; Cravedi, Paolo; Katari, Ravi; Maghsoudlou, Panagiotis; Fallas, Mario Enrique Alvarez; Sordi, Valeria; Citro, Antonio; Purroy, Carolina; Niu, Guoguang; McQuilling, John P; Sittadjody, Sivanandane; Farney, Alan C; Iskandar, Samy S; Zambon, Joao P; Rogers, Jeffrey; Stratta, Robert J; Opara, Emmanuel C; Piemonti, Lorenzo; Furdui, Cristina M; Soker, Shay; De Coppi, Paolo; Orlando, Giuseppe

    2016-07-01

    Our study aims at producing acellular extracellular matrix scaffolds from the human pancreas (hpaECMs) as a first critical step toward the production of a new-generation, fully human-derived bioartificial endocrine pancreas. In this bioartificial endocrine pancreas, the hardware will be represented by hpaECMs, whereas the software will consist in the cellular compartment generated from patient's own cells. Extracellular matrix (ECM)-based scaffolds obtained through the decellularization of native organs have become the favored platform in the field of complex organ bioengineering. However, the paradigm is now switching from the porcine to the human model. To achieve our goal, human pancreata were decellularized with Triton-based solution and thoroughly characterized. Primary endpoints were complete cell and DNA clearance, preservation of ECM components, growth factors and stiffness, ability to induce angiogenesis, conservation of the framework of the innate vasculature, and immunogenicity. Secondary endpoint was hpaECMs' ability to sustain growth and function of human islet and human primary pancreatic endothelial cells. Results show that hpaECMs can be successfully and consistently produced from human pancreata and maintain their innate molecular and spatial framework and stiffness, and vital growth factors. Importantly, hpaECMs inhibit human naïve CD4 T-cell expansion in response to polyclonal stimuli by inducing their apoptosis and promoting their conversion into regulatory T cells. hpaECMs are cytocompatible and supportive of representative pancreatic cell types. We, therefore, conclude that hpaECMs has the potential to become an ideal platform for investigations aiming at the manufacturing of a regenerative medicine-inspired bioartificial endocrine pancreas.

  5. 烧伤变性脱细胞真皮基质可再生利用的实验研究%Experimental study on the recycling of denatured acellular dermal matrix after burn

    Institute of Scientific and Technical Information of China (English)

    王晓川; 李川; 单菲; 王文婷; 朱旭国; 姜笃银

    2012-01-01

    Objective To explore the feasibility of burn denatured acellular dermal matrix (DADM) as dermal substitute in repairing wounds. Methods ( 1 ) Nine Wistar rats received a deep partial-thickness scald on the back.Full-thickness wounded skin was collected on post scald day (PBD) 1,2,and 3 (with 3 rats at each time point),and it was treated with 2.5 g/L trypsin/0.5% Triton X-100 to remove cells to prepare DADM,respectively called DADM-1 d,DADM-2 d,and DADM-3 d.Another 3 rats without scald injury were treated with the same method as above to prepare acellular dermal matrix (ADM)to serve as control.Gross and histological observations and microbiological and biomechanical tests,including ultimate tensile strength,maximum tension,stretched length at breaking,stress-strain relationship,were conducted for the resulting ADM and DADM.(2) Another 64 rats were divided into ADM group and DADM-1 d,DADM-2 d,and DADM-3 d groups according to the random number table,with 16 rats in each group.A skin flap in size of 2.0 cm× 1.8 cm was raised on the back of each rat.The above-mentioned ADM,DADM-1 d,DADM-2 d,and DADM-3 d were cut into pieces in the size of 1.8 cm × 1.5 cm,and they were respectively implanted under the skin flaps of rats in corresponding group.At post surgery week (PSW) 1,3,5,or 9,4 rats in each group were used to observe wound healing condition and change in implants with naked eye,and histological observation of the implants was conducted.Data were processed with one-way analysis of variance and t test. Results ( 1 ) The freshly prepared DADM was milky white,soft in texture with flexibility,but poor in elasticity as compared with ADM.No epithelial structure or cellular component was observed in ADM or DADM under light microscope.Collagen fibers of DADM were seen to be thickened unevenly and arranged in disorder and eosinophilic.All microbiological results of DADM were negative.There was no statistically significant difference among DADM-1 d,DADM-2 d,and DADM-3 d in

  6. 脱细胞真皮基质治疗伴高位盲瘘的复杂性肛瘘疗效观察(附39例报告)%Acellular Dermal Matrix treatment of high blind fistula of 39 Cases

    Institute of Scientific and Technical Information of China (English)

    田颖; 于洪顺; 秦澎湃; 王彦芳; 田磊; 葛强; 刘智永

    2012-01-01

    目的 探讨高位盲瘘的微创治疗方法,研究脱细胞真皮基质在高位盲瘘治疗中的应用价值.方法 39例高位盲瘘患者,手术分2期进行,经括约肌或括约肌间肛瘘合并高位盲瘘Ⅰ期齿线下瘘管切开、内口挂线并高位盲瘘旷置引流,括约肌上肛瘘切开内口并高位盲瘘旷置引流;Ⅱ期高位盲瘘脱细胞真皮基质填塞治疗.观察Ⅱ期手术时间、术中出血、术后疼痛、住院总天数、住院总费用及复发率等临床及相关指标.结果 39例患者中有26例获得Ⅰ期治愈,13例患者治疗失败,改行肛瘘切开挂线术后痊愈.高位盲瘘脱细胞真皮填塞术手术治愈率66.7%.括约肌上并发高位盲瘘治愈2例,治愈率100%,经括约肌并发高位盲瘘治愈6例,治愈率50%,括约肌间并发高位盲瘘治愈18例,治愈率72%.结论 应用脱细胞真皮基质材料治疗高位盲瘘具有损伤小、愈合时间短、肛门失禁率低、外形保留好等优势,值得进一步推广.%Objective To investigate the minimally invasive treatment of high blind fistula, acellular dermal matrix in the high blind fistula treatment value. Methods high blind fistula two cases: 39 cases of patients with high blind fistula, anal sphincter, sphincter anal fistula complicated by high blind fistula of 12 cases, anal sphincter between concurrent high blind fistula of 25 cases. Operation in two, fistulotomy anal sphincter or sphincter merge high blind fistula I of the dentate line, inside the mouth hung high blind fistula exclusion drainage, anal sphincter incision inside the mouth and the high blind fistula exclusion fistula drainage; II high blind acellular dermal matrix filling treatment observation period II surgery time, blood loss, postoperative pain, total hospital days, hospital clinical and related indicators of the total cost and the relapse rate. Results 39 patients, 26 cases of a cure, 13 cases of treatment failure in patients diverted a-nal incision

  7. Evaluation of the biocompatibility and cell segregation performance of acellular dermal matrix as barrier membrane on guided tissue regeneration in vitro%脱细胞真皮基质作为屏障膜的细胞相容性及细胞封闭性的体外研究

    Institute of Scientific and Technical Information of China (English)

    陈武; 王韦玮; 时新站; 陈宁

    2013-01-01

    目的:研究脱细胞真皮基质(acellular dermal matrix,ADM)对人牙周膜细胞增殖及上皮细胞的封闭性能的影响,评估其作为引导组织再生屏障膜的可行性.方法:取因正畸需要拔除的新鲜第一前磨牙,刮取根中1/3牙周膜组织,组织块法进行人牙周膜细胞(human periodontal ligament cells,HPDLCs)的原代培养.将ADM膜、膨体聚四氟乙烯(expanded polytetrafluoroethylene,e-PTFE)膜预处理后与HPDLCs共培养,MTT法检测1、3、5、7d的细胞增殖活性.将Tca8113细胞接种于膜材料一侧表面,培养5、10d后,采用DAPI细胞核染色,在荧光显微镜下观察细胞在膜材料两面的分布情况,接种细胞面记为ADM组与e-PTFE组,另一面记为ADM’组与e-PTFE’组.数据采用SPSS 13.0软件包进行t检验.结果:3、5、7d时,ADM组和空白对照组的OD值显著高于e-PTFE组(P<0.05),ADM组与空白对照组的OD值差异无显著性(P>0.05).ADM组与ADM’组、e-PTFE组与e-PTFE’组在5、10d时细胞计数均有显著差异(P<0.05);ADM’组与e-PTFE’组在5、10 d时细胞计数无显著差异(P>0.05).结论:ADM膜比e-PTFE更有利于HPDLCs的增殖,且两者对上皮细胞的封闭作用相似.与e-PTFE相比,ADM更适合用于引导牙周组织再生术.%PURPOSE:To investigate the proliferation of human periodontal ligament cell on acellular dermal matrix (ADM) and the epithelial cell segregation performance of ADM and evaluate the feasibility of ADM as barrier membrane of guided tissue regeneration.METHODS:Human periodontal ligament cells (HPDLCs) of the 3rd to 5th passage were seeded onto 96-well plates(with ADM and e-PTFE inside) with 2000 cells per well.The cells were cultured in Dulbecco's modified eagle medium (DMEM).The MTT colorimetric assay method was performed at day 1,3,5 and 7 after incubation.The optical density (OD) of each well was measured spectrophotometrically at 490 nm to monitor effects on cell proliferation.The data was analyzed using

  8. Clinical study of nanosilver/porcine acellular dermal matrix dressing in the treatment of superficial second degree burn wounds%纳米银-猪脱细胞真皮基质敷料治疗浅Ⅱ度烧伤创面的临床观察

    Institute of Scientific and Technical Information of China (English)

    胡晓文; 郝天智; 张华

    2016-01-01

    天的创面愈合率明显优于其余两组,差异均有统计学意义( P值均小于0.05);创面愈合时间显著短于其余两组,比较差异均有统计学意义(P值均小于0.05)。结论纳米银-猪脱细胞真皮基质敷料具有抗感染、促进创面愈合及减轻换药痛觉的作用。%Objective To observe the clinical effect of nanosilver/porcine acellular dermal matrix dressing on superficial second degree burn wounds. Methods From January 2014 to December 2015,90 patients with superficial second degree burn were treated and observed in the Department of Burns and Plastics Surgery, General Hospital of Beijing Military Region. According to the order of admission and random number table, the patients were randomly divided into 3 groups, nanosilver dressing group, porcine acellular dermal matrix dressing group and nanosilver/porcine acellular dermal matrix dressing group, each group of 30 patients. On the day of admission, the wounds areas were calculated by taking pictures and the wound secretion was taken for bacterial culture by using the throat swab, burn wounds were treated with debridement, then apply nanosilver dressing, porcine acellular dermal matrix dressing and nanosilver/porcine acellular dermal matrix dressing on the wounds. On the 5th day after treatment , the wound secretion was taken for bacterial culture by using the throat swab again. Pain score was assessed by asking and observing the changes of pain in patients after dressing change. On the 7th day after treatment, the wounds areas were calculated by taking pictures. The wound healing rate was calculated. Wound healing time was recorded. The data were compared by using one-way ANOVA test, SNK-q test and Chi-square test. Results On the 5th day after treatment, the wound bacterial culture results of nanosilver dressing group, porcine acellular dermal matrix dressing group and nanosilver/porcine acellular dermal matrix dressing group were 2(6.6%),9(30. 0%),1(3. 3%),there were

  9. Fibre-matrix interaction in the human annulus fibrosus.

    Science.gov (United States)

    Guo, Zaoyang; Shi, Xiaohao; Peng, Xiongqi; Caner, Ferhun

    2012-01-01

    Although the mechanical behaviour of the human annulus fibrosus has been extensively studied, the interaction between the collagen fibres and the ground matrix has not been well understood and is therefore ignored by most constitutive models. The objective of this study is to identify the significance of the fibre-matrix interaction in the human annulus fibrosus by careful investigation of the experimental data, the theoretical constitutive models, and the numerical simulation results in the literature. Based on the experimental results from biaxial and uniaxial tests, it is shown that the mechanical behaviour of the matrix can be well simulated by an incompressible neo-Hookean type model, but the effective stiffness of the matrix depends on fibre stretch ratio, which can only be explained by fibre-matrix interaction. Furthermore, we find that this interaction takes place anisotropically between the matrix and the fibres distributed in different proportions in different directions. The dependence of the tangent stiffness of the matrix on the first invariant of the deformation tensor can also be explained by this fibre orientation dispersion.

  10. Update of human and mouse matrix metalloproteinase families

    OpenAIRE

    Jackson Brian C; Nebert Daniel W; Vasiliou Vasilis

    2010-01-01

    Abstract Matrix metalloproteinases (MMPs) are a family of zinc proteases that degrade most of the components of the extracellular matrix (ECM). MMPs also have a number of non-traditional roles in processing factors related to cell growth/proliferation, inflammation and more. There are 23 human MMPs and 23 mouse MMPs, most of which share orthology among most vertebrates; other examples have been found in invertebrates and plants. MMPs are named in order of discovery, but also have been grouped...

  11. Study of compatibility of acellular cartilage extracellular matrix-derived porous scaffolds with sheep nucleus pulposus cells%软骨脱细胞细胞外基质多孔支架与山羊髓核细胞生物相容性研究

    Institute of Scientific and Technical Information of China (English)

    伍耀宏; 徐宝山; 杨强; 李秀兰; 张杨; 夏群; 张春秋; 许海委

    2013-01-01

    Objective To study the compatibility of acellular cartilage extracellular matrix-derived porous scaffolds with sheep nucleus pulposus cells.Methods Articular cartilage derived from pigs was physically shattered and decellularized,and then made into porous scaffolds with freeze-drying techniques.Nucleus pulposus cells were isolated from the goat lumbar intervertebral disc,and P1 generation were obtained after culturing.The toxicity of leaching liquor from scaffolds was tested by MTT assay.The cells were seeded onto scaffolds with a density of 5 x 106/ml and cultured for 48h in vitro,activity and adhesion for cells on scaffolds were evaluated by inverted microscope,HE staining,LIVE/DEAD staining and scanning electron microscopy.Results Acellular cartilage extracellular matrix-derived porous scaffolds were smooth and transparent,isolated nucleus pulposus cells showed typical chondrocyte-like morphology.MTT assay demonstrated that proliferation among the groups has no significant difference(P>0.05).Cells showed spherical or short-spindle morphology and attached to the scaffolds evenly under the inverted microscope and scanning electron microscopy,and HE staining confirmed the even attachment of the cells.All the cells showed green fluorescence (live cells) while no red fluorescence (dead cells) was observed after staining with LIVE/DEAD dye.Conclusion The acellular cartilage extracellular matrix-derived porous scaffolds can be used as the nucleus pulposus tissue for sharing similar extracellular matrix composition with nucleus pulposus tissue and possess good cell compatibility with the sheep nucleus pulposus cells.%目的 制备软骨脱细胞细胞外基质多孔支架,并探讨其与山羊髓核细胞的生物相容性.方法 猪关节软骨经研磨、脱细胞、冷冻干燥技术等处理制成三维多孔支架;从山羊腰椎间盘中分离出髓核细胞,培养后获取P1代细胞;四甲基偶氮唑蓝(MTT)检测支架浸提

  12. Characterisation of the circulating acellular proteome of healthy sheep using LC-MS/MS-based proteomics analysis of serum.

    Science.gov (United States)

    Chemonges, Saul; Gupta, Rajesh; Mills, Paul C; Kopp, Steven R; Sadowski, Pawel

    2016-01-01

    Unlike humans, there is currently no publicly available reference mass spectrometry-based circulating acellular proteome data for sheep, limiting the analysis and interpretation of a range of physiological changes and disease states. The objective of this study was to develop a robust and comprehensive method to characterise the circulating acellular proteome in ovine serum. Serum samples from healthy sheep were subjected to shotgun proteomic analysis using nano liquid chromatography nano electrospray ionisation tandem mass spectrometry (nanoLC-nanoESI-MS/MS) on a quadrupole time-of-flight instrument (TripleTOF® 5600+, SCIEX). Proteins were identified using ProteinPilot™ (SCIEX) and Mascot (Matrix Science) software based on a minimum of two unmodified highly scoring unique peptides per protein at a false discovery rate (FDR) of 1% software by searching a subset of the Universal Protein Resource Knowledgebase (UniProtKB) database (http://www.uniprot.org). PeptideShaker (CompOmics, VIB-UGent) searches were used to validate protein identifications from ProteinPilot™ and Mascot. ProteinPilot™ and Mascot identified 245 and 379 protein groups (IDs), respectively, and PeptideShaker validated 133 protein IDs from the entire dataset. Since Mascot software is considered the industry standard and identified the most proteins, these were analysed using the Protein ANalysis THrough Evolutionary Relationships (PANTHER) classification tool revealing the association of 349 genes with 127 protein pathway hits. These data are available via ProteomeXchange with identifier PXD004989. These results demonstrated for the first time the feasibility of characterising the ovine circulating acellular proteome using nanoLC-nanoESI-MS/MS. This peptide spectral data contributes to a protein library that can be used to identify a wide range of proteins in ovine serum.

  13. Morphology of spinal cord extracellular matrixderived acellular scaffolds fabricated in rats

    Institute of Scientific and Technical Information of China (English)

    Wenhua Yin; Kaiwu Lu; Dadi Jin

    2011-01-01

    Acellular peripheral allograft scaffolds can be fabricated using chemical extraction techniques, but methods for producing acellular scaffold derived from spinal cord tissue are not currently available.The present study demonstrated that chemical extraction using Triton X-100 and sodium deoxycholate could be used to completely remove the cells, axons and neural sheaths in spinal cord extracellular matrix-derived scaffolds. The matrix fibers were longitudinally arranged in a wave-like formation, and were connected by fiber junctions. Lattice-shaped fiber cages appeared and developed into bone trabecula-like changes. The natural structure of matrix fibers in the scaffolds was maintained; this helps to guide the differentiation and migration of implanted stem cells. Decellularized spinal cord extracellular matrix-derived scaffolds can provide an ideal substance for fabricating tissue-engineered spinal cord.

  14. 脱细胞真皮基质对骨质疏松大鼠骨缺损愈合影响%Effect of acellular dermal matrix on osteoporosis rats bone defect healing

    Institute of Scientific and Technical Information of China (English)

    王晓晗; 赵志国; 王智明; 王文茉; 张力平

    2016-01-01

    Objective To observe the characteristics and capabilities of acellular dermal matrix( ADM) in osteoporosis rats cranial parietal bone defect repair guided bone regeneration( GBR) ,to explore the biocompatibility and effects on bone re-generation. Methods A total of 26 SD female rats were randomly divided into the control group(Sham group:n=13)and the ovariectomized group(VOX group:n=13). Conventional breeding for 3 months after the surgery,after the success of the building,in skull of rats,there were 2 defective holes with 5 mm preparation on both sides of central line,one side was cov-ered with ADM,the other side was control blank( CK) . In Sham group,the CK side was Group A,the ADM cover side was Group B. In the CK side in OVX group was Group C,the ADM cover side was Group D. In 6 and 12 weeks postoperatively, the clinical features such as the bone defect healing,bone tissue HE and masson trichromatic dyeing,new bone lengths were compared,the mineralization rate,immunohistochemical method was to detect callus osteocalcin expression in different peri-ods. Results Among the gross observation experimental animals,there were 2 deaths caused by bowel bilges gas in. Other animals healed within a week without infection and wound dehiscence,visible sutures were not fallen off. 6 weeks after sur-gery,the blank defects naked eye obvious difference between the two groups,the defect area was covered with transparency, defect edge was clear. ADM cover side was with visible white ADM,defect edge was clear. After 12 weeks,there was no na-ked eye obvious difference between two groups of blank defect,the defect area was covered with transparency,defect edge was clear. ADM cover side with visible white ADM film was thinner,harder to hit,defect edge was obtuse. The tissue mor-phology observation 6 weeks when two groups of new bone gap defect was not obvious,the broken end by fibrous tissue pack-age. Sham group of bone defect end osteogenesis was dense, ADM retained membrane

  15. 脱细胞真皮基质对扩张器/假体乳房再造并发症影响的Meta分析%The impact of acellular dermal matrix on complications of breast reconstruction using tissue expander/implant: a Meta-analysis

    Institute of Scientific and Technical Information of China (English)

    董洁; 吴小蔚; 田方兴

    2013-01-01

    Objective To analyze the effect of acellular dermal matrix (ADM) on complications of breast reconstruction using tissue expander/implant,and to offer preliminary evidences for ADM clinical application.Methods Articles published from Jan.2010 to Oct.2012 were searched in Pubmed,EMbase,Science Direct and CNKI database.Literatures were filtrated according to inclusive criteria.Values were extracted from included literatures; factors regarding complications were collected.Metaanalysis was performed with Stata 12.0.Results 10 researches were included.Comparing to control group,the pooled odds ratio (OR) of overall complications,infections,hematomas/seromas,explantations are 1.51(P=0.038),1.91(P=0.032),1.80(P=0.005) and 2.37 (P=0.138) in ADM group respectively.Conclusions In breast reconstruction using tissue expander/implant,ADM increases the occurrence of hematomas/saromas as well as risks of infections and overall complications.%目的 对在扩张器/假体乳房再造术中应用脱细胞真皮基质(acellular dermal matrix,ADM)是否增加术后并发症进行探讨,以为临床应用提供初步依据.方法 计算机检索2010年1月至2012年10月Pubmed、EMbase、Science Direct、中国生物医学文献数据库和中国期刊网全文数据库中发表的文献,设定文献纳入条件,对符合条件的文献进行并发症相关数据导出,然后使用Stata 12.0进行Meta分析.结果 共有10篇文献被纳入分析.与对照组相比,ADM组总并发症、感染,血肿/血清肿及扩张器/假体取出发生率的比值比(OR)分别为1.51(P =0.038)、1.91(P=0.032)、1.80(P =0.005)和2.37(P =0.138).结论 在扩张器/假体的乳房再造术中,使用ADM对血肿/血清肿的发生有促进作用,并有增加感染率及总并发症发生率的趋势.

  16. Clinical Evaluation of Heterogeneous Acellular Dermal Matrix for Guided Tissue Regeneration in Periodontal Disease%异种脱细胞真皮基质膜在引导牙周组织再生中应用的临床效果评价

    Institute of Scientific and Technical Information of China (English)

    章立群; 邓碧霞; 谢安琪; 孙辉

    2012-01-01

    Objective: To explore the clinical effectiveness of heterogeneous acellular dermal matrix( AMD) membrane for guided tissue regeneration in treatment of periodontal defects. Methods: 30 periodontal defects were randomly assigned into two treatments; 20 for testing group using a combination of AMD and coralline hydroxyapatite (HA) grafting and 10 for control group using HA grafting alone. After six months of surgery, the change of clinical index and radiographic alveolar bone level were compared statistically. Results: The reduction of pocket probing depth and clinical attachment loss and increasing level of alveolar bone of testing group were all higher than those in control group while the recession quantity was lower than that in control group(P<0. 05). Conclusion: Heterogeneous acellular dermal matrix membrane is effective for the treatment of periodontal defects.%目的:评价异种脱细胞真皮基质膜在引导牙周组织再生中应用的临床效果.方法:30例牙周缺损区患牙随机分两组:实验组20例,异种脱细胞真皮基质膜和羟基磷灰石修复;对照组10例仅羟基磷灰石修复.统计学比较6个月时两组各项临床指标和牙槽骨水平改变.结果:实验组牙周袋深度、临床附着丧失的减少量和牙槽骨水平的增加量均较对照组高(P<0.05),牙龈退缩量低于对照组(P<0.05).结论:异种脱细胞真皮基质膜应用于引导牙周组织再生有较好的临床效果,值得推广.

  17. Characterisation of the circulating acellular proteome of healthy sheep using LC-MS/MS-based proteomics analysis of serum

    OpenAIRE

    Chemonges, Saul; Gupta, Rajesh; Mills, Paul C.; Steven R. Kopp; Sadowski, Pawel

    2017-01-01

    Background Unlike humans, there is currently no publicly available reference mass spectrometry-based circulating acellular proteome data for sheep, limiting the analysis and interpretation of a range of physiological changes and disease states. The objective of this study was to develop a robust and comprehensive method to characterise the circulating acellular proteome in ovine serum. Methods Serum samples from healthy sheep were subjected to shotgun proteomic analysis using nano liquid chro...

  18. STUDY ON NUCLEAR MATRIX PROTEINS FROM HUMAN BREAST CARCINOMA

    Institute of Scientific and Technical Information of China (English)

    HE Qian; ZHANG Shu-qun; CHU Yong-lie; JIA Xiao-li; JIANG Jian-tao

    2009-01-01

    Objective To investigate the marker protein of human breast carcinoma from nuclear matrix proteins (NMPs).Methods NMPs were injected subcutaneously into rabbit to get antiserum, which was used to detect the NMPs specificity for breast carcinoma.Results There was an apparent positive band (100kD) in the NMPs of breast carcinoma, which did not exist in normal breast and other tumors that were detected.Conclusion One or one group of 100kD NMPs were found to be related to human breast carcinoma, which may be involved in the carcinogenesis and development of human breast carcinoma and valuable for breast carcinoma diagnosis.

  19. Acellular dermis-assisted breast reconstruction.

    Science.gov (United States)

    Spear, S L; Parikh, P M; Reisin, E; Menon, N G

    2008-05-01

    In 2004, the authors reported their findings with placement of tissue expanders for breast reconstruction in the partial submuscular position, the equivalent of the "dual-plane" technique for breast augmentation. Limitations with subpectoral expander placement include difficulty controlling the lower pole of the pocket during expansion, unprotected device coverage by a thin inferior mastectomy flap, possible effacement of the inframammary fold, and limited control over the superior migration of the pectoralis major muscle. This study aimed to examine the safety and efficacy of an acellular dermal sling in providing inferolateral support to the device during immediate breast reconstruction and expansion. This study prospectively investigated 58 breasts of 43 consecutive women who underwent immediate breast reconstruction with tissue expanders and acellular dermis. After completion of adjuvant therapy and expansion, the devices were exchanged for implants. The patients were tracked through January, 2007. The study parameters included demographic information, oncologic data, complications, and aesthetic outcomes. The mean time required to complete reconstruction was 8.6 months. The overall complication rate after expander/acellular dermis placement was 12%, whereas the complication rate after exchange to implants was 2.2%. The aesthetic outcome for reconstructed breasts did not differ significantly from that for the control subjects who had no surgery. Acellular dermis appears to be a useful adjunct in immediate prosthetic breast reconstruction. Acellular dermis-assisted breast reconstruction has a low complication rate, helps to reconstruct an aesthetically pleasing breast, and facilitates expeditious completion of the reconstruction.

  20. Replacement of animal-derived collagen matrix by human fibroblast-derived dermal matrix for human skin equivalent products.

    Science.gov (United States)

    El Ghalbzouri, Abdoelwaheb; Commandeur, Suzan; Rietveld, Marion H; Mulder, Aat A; Willemze, Rein

    2009-01-01

    Reconstructed human skin equivalents (HSEs) are representative models of human skin and widely used for research purposes and clinical applications. Traditional methods to generate HSEs are based on the seeding of human keratinocytes onto three-dimensional human fibroblast-populated non-human collagen matrices. Current HSEs have a limited lifespan of approximately 8 weeks, rendering them unsuitable for long-term studies. Here we present a new generation of HSEs being fully composed of human components and which can be cultured up to 20 weeks. This model is generated on a primary human fibroblast-derived dermal matrix. Pro-collagen type I secretion by human fibroblasts stabilized during long-term culture, providing a continuous and functional human dermal matrix. In contrast to rat-tail collagen-based HSEs, the present fibroblast-derived matrix-based HSEs contain more continuity in the number of viable cell layers in long-term cultures. In addition, these new skin models exhibit normal differentiation and proliferation, based on expression of K10/K15, and K16/K17, respectively. Detection of collagen types IV and VII and laminin 332 was confined to the epidermal-dermal junction, as in native skin. The presence of hemidesmosomes and anchoring fibrils was demonstrated by electron microscopy. Finally, we show that the presented HSE contained a higher concentration of the normal moisturizing factor compared to rat-tail collagen-based skin models, providing a further representation of functional normal human skin in vitro. This study, therefore, demonstrates the role of the dermal microenvironment on epidermal regeneration and lifespan in vitro.

  1. Histological changes of acellular dermal matrix used as guided tissue regeneration barrier membrane in vivo%脱细胞真皮基质作为引导组织再生屏障膜在体内的组织学变化研究

    Institute of Scientific and Technical Information of China (English)

    王乾锋; 刘宏伟; 郑秋林

    2010-01-01

    目的 观察脱细胞真皮基质(acellular dermal matrix,ADM)作为引导组织再生(guided tissue regeneration,GTR)屏障膜时的体内组织学变化,探索其作为GTR屏障膜的可行性.方法 在兔下颌前磨牙颊侧根面形成一开窗型牙周缺损模型,将ADM作为GTR屏障膜覆盖和固定于缺损区表面,观察术后4周和8周时ADM的降解、血管化、炎性反应等组织学变化情况.结果 术后4周时,ADM无明显降解,基本保持其原来完整结构;8周时,ADM出现轻中度降解,但结缔组织尚未突破整层ADM.结论 ADM可以作为GTR的屏障膜.

  2. 无细胞异体真皮基质在烧伤后整形患者功能部位的应用%The application of acellular dermal matrix allograft in functional position of patients with post- burns plastic operation

    Institute of Scientific and Technical Information of China (English)

    姜笃银; 杨银辉; 张玮; 付小兵

    2003-01-01

    AIM:To investigate the effect of allogeneic acellular dermal matrix(ADM) on cograft in joint functional positions of patients with post burn plastic operation. METHODS:9 patients with hypertrophic scar and joint dysfunction after severe burns were used. After pre treating with trypsin and TritonX 100, 13 reticulated ADM were overlapped with autogenous ultrathin split thickness skin grafts(USTS), and were transplanted to the scar excision wounds in the joints of four limbs at the same time. The neighbouring autogenous thin split thickness skin grafts(TSTS) were used as control.RESULTS:The composite skin grafts as well as the controls were all survived. The rejection and hypertrophic scars were not found during (1- 5) years follow up studies. The appearance, fiber and function of composite skin grafts were near to normal skins. CONCLUSION:The ADM could be used to joint functional positions of patients with post burn hypertrophic scars and could produce satisfactory plastic results as dermal substitute.

  3. Factors Involved in Extracellular Matrix Turnover in Human Derived Cardiomyocytes

    Directory of Open Access Journals (Sweden)

    Gregori Casals

    2013-11-01

    Full Text Available Background: The molecular mechanisms by which myocardial ischemia translates into ventricular remodeling remain unclear. Methods: We investigated whether hypoxia and proinflammatory cytokines are specific inducers of remodeling signals in an in vitro model of cultured adult human ventricular myocytes (AC16 cells. Results:Hypoxia modified the ratio of matrix remodeling factors by increasing the aminoterminal propeptide of type III procollagen (PIIINP and reducing tissue inhibitor of matrix metalloproteinase type 1 (TIMP-1 secretion in AC16 cells. These effects, however, were not associated with either modifications in expression of matrix metalloproteinase type 2, collagen-I or metalloproteinase activity. Hypoxia does, actually increase the production of the cardiac antifibrogenic growth factors, Apelin and VEGF, through an Hypoxia Inducible Factor type 1-dependent mechanism. Concerning proinflammatory signaling pathways, IL1β emerged as a powerful inducer of matrix turnover, since it significantly enhanced PIIINP, TIMP-1 and hyaluronic acid production and increased metalloproteinase activity. In contrast, TNFα did not modify matrix turnover but markedly induced the production of Apelin and VEGF. Conclusion: Hypoxia and increased TNFα activity likely exert cardioprotective actions by activating the cardiac antifibrogenic factors Apelin and VEGF. In contrast, IL1β is a strong promoter of interstitial collagen remodeling that may contribute to ventricular dilation and heart failure in the ischemic myocardium.

  4. Identification of osteopontin in human dental calculus matrix.

    Science.gov (United States)

    Kido, J; Kasahara, C; Ohishi, K; Nishikawa, S; Ishida, H; Yamashita, K; Kitamura, S; Kohri, K; Nagata, T

    1995-10-01

    Osteopontin is a prominent non-collagenous component of bone matrix, although it is expressed in several other tissues. Recently, osteopontin was reported to be involved in urinary stone formation and atherosclerotic lesions of the aorta, suggesting that it may be a key protein associated with these types of pathological mineralization. In this study, whether or not human dental calculus contains osteopontin was investigated by immunoblotting and immunohistochemical analyses. After extraction of calculus proteins with EDTA and separation of the proteins by electrophoresis, immunoblotting analysis revealed the presence of osteopontin. Two forms of osteopontin appeared at 61 and 68 kDa on 10% polyacrylamide gel and the proteins were digested with thrombin, a highly specific protease. Moreover, immunohistochemical analysis revealed that osteopontin was localized in dental calculus adherent to tooth roots. These findings indicate that osteopontin is, in fact, present in human dental calculus and may be involved in calculus formation as the stone matrix.

  5. [STUDY ON MODIFICATION OF BIOMATERIALS OF ACELLULAR BOVINE PERICARDIUM WITH DIFFERENT CROSSLINKING REAGENTS].

    Science.gov (United States)

    Xiao, Hongtao; Tian, Shemin; Zha, Xinjian; Wei, Ying; Huang, Hongjun; Li, Yun; Yang, Huanna; Xia, Chengde; Niu, Xihua

    2015-10-01

    To investigate the effects of modification of acellular bovine pericardium with 1-ethyl-3-(3- dinethylami-nopropyl) carbodimide (EDC)/N-hydroxysuccininide (NHS) or genipin and find out the best crosslinking reagent. The cellular components of the bovine pericardiums were removed. The effects of decellularization were tested by HE staining. The acellular bovine pericardiums were crosslinked with EDC/NHS (EDC/NHS group) or genipin (genipin group). The properties of the crosslinked acellular matrix were evaluated by scanning electron microscope (SEM), matrix thickness, crosslinking index, mechanical property, denaturation temperature, enzymatic degradation, and cytotoxicity test before and after the crosslinking. Acellular bovine pericardium (ABP group) or normal bovine pericardium (control group) were harvested as controls. SEM showed that collagen fibers were reticulated in bovine pericardial tissues after crosslinked by EDC/NHS or genipin, and relative aperture of the collagen fiber was from 10 to 20 μm. The thickness and denaturation temperature of the scaffolds were increased significantly after crosslinking with EDC/NHS or genipin (P 0.05). The difference had no statistical significance in crosslinking index between EDC/NHS group and genipin group (t = 0.205, P = 0.218). The degradation rate in EDC/NHS group and genipin group was significantly lower than that in ABP group and control group (P 0.05). The break elongation in EDC/NHS group and genipin group were significantly increased than those in ABP group and control group (P 0.05). Cytotoxicity of genipin crosslinked tissue (grade 1) were much lower than that of EDC/NHS (grade 2) at 5 days. Acellular bovine pericardium crosslinked with genipin has better biocompatibility than EDC/NHS.

  6. New Insights on the Composition and the Structure of the Acellular Extrinsic Fiber Cementum by Raman Analysis

    Science.gov (United States)

    Colard, Thomas; Falgayrac, Guillaume; Bertrand, Benoit; Naji, Stephan; Devos, Olivier; Balsack, Clara; Delannoy, Yann; Penel, Guillaume

    2016-01-01

    Acellular extrinsic fiber cementum is a mineralized tissue that covers the cervical half of the tooth root surface. It contains mainly extrinsic or Sharpey’s fibers that run perpendicular to the root surface to anchor the tooth via the periodontal ligament. Acellular cementum is continuously and slowly produced throughout life and exhibits an alternating bright and dark pattern under light microscopy. However, although a better understanding of the structural background of acellular cementum is relevant to many fields, such as cementochronology, periodontology and tissue engineering, acellular cementum remains rarely studied and poorly understood. In this work, we studied the acellular cementum at the incremental line scale of five human mandibular canines using polarized Raman spectroscopy. We provided Raman imaging analysis and polarized acquisitions as a function of the angular orientation of the sample. The results showed that mineral crystals were always parallel to collagen fibrils, and at a larger scale, we proposed an organizational model in which we found radial collagen fibers, “orthogonal” to the cementum surface, and “non-orthogonal” fibers, which consist of branching and bending radial fibers. Concerning the alternating pattern, we observed that the dark lines corresponded to smaller, more mineralized and probably more organized bands, which is consistent with the zoological assumption that incremental lines are produced during a winter rest period of acellular cementum growth. PMID:27936010

  7. New Insights on the Composition and the Structure of the Acellular Extrinsic Fiber Cementum by Raman Analysis.

    Science.gov (United States)

    Colard, Thomas; Falgayrac, Guillaume; Bertrand, Benoit; Naji, Stephan; Devos, Olivier; Balsack, Clara; Delannoy, Yann; Penel, Guillaume

    2016-01-01

    Acellular extrinsic fiber cementum is a mineralized tissue that covers the cervical half of the tooth root surface. It contains mainly extrinsic or Sharpey's fibers that run perpendicular to the root surface to anchor the tooth via the periodontal ligament. Acellular cementum is continuously and slowly produced throughout life and exhibits an alternating bright and dark pattern under light microscopy. However, although a better understanding of the structural background of acellular cementum is relevant to many fields, such as cementochronology, periodontology and tissue engineering, acellular cementum remains rarely studied and poorly understood. In this work, we studied the acellular cementum at the incremental line scale of five human mandibular canines using polarized Raman spectroscopy. We provided Raman imaging analysis and polarized acquisitions as a function of the angular orientation of the sample. The results showed that mineral crystals were always parallel to collagen fibrils, and at a larger scale, we proposed an organizational model in which we found radial collagen fibers, "orthogonal" to the cementum surface, and "non-orthogonal" fibers, which consist of branching and bending radial fibers. Concerning the alternating pattern, we observed that the dark lines corresponded to smaller, more mineralized and probably more organized bands, which is consistent with the zoological assumption that incremental lines are produced during a winter rest period of acellular cementum growth.

  8. Preparation and characterization of an acellular bovine pericardium intended for manufacture of valve bioprostheses.

    Science.gov (United States)

    Goissis, Gilberto; Giglioti, Aparecida de Fátima; Braile, Domingo Marcolino

    2011-05-01

    Major problems with biological heart valves post-implantation are associated with progressive structural deterioration and calcification attributed to glutaraldehyde processing, dead cells, and cell fragments present in the native tissue. In spite of these problems, glutaraldehyde still is the reagent of choice. The results with acellular matrix xenograft usually prepared by detergent treatment in association with enzymes are rather conflicting because while preserving mechanical properties, tissue morphology and collagen structure are process dependent. This work describes a chemical approach for the preparation of an acellular bovine pericardium matrix intended for the manufacture of heart valve bioprostheses. Cell removal was performed by an alkaline extraction in the presence of calcium salts for periods ranging from 6 to 48 h. The results showed that cell removal was achieved after 12 h, with swelling and negative charge increasing with processing time. Nevertheless, collagen fibril structure, ability to form fibrils, and stability to collagenase were progressive after 24-h processing. There was no denaturation of the collagen matrix. A process is described for the preparation of acellular bovine pericardium matrices with preserved fibril structure and morphology for the manufacture of cardiac valve bioprostheses and may be used in other applications for tissue reconstruction. © 2011, Copyright the Authors. Artificial Organs © 2011, International Center for Artificial Organs and Transplantation and Wiley Periodicals, Inc.

  9. Decalcificated human dentin matrix in autogenous repair of skull defects

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    @@In the management of traumatic skull defect, the classical treatment has usually been adopted, i.e.,primary debridement and secondary repair of bone defect, especially in cases of open lacerated skull fracture. 1 In general, the use of prosthetic material in repair is often not so satisfactory either in China or abroad. Decalcificated human dentin matrix (DHDM)has been used in autogenous repair of traumatic skull defect in primary operation and a good curative effect has been gained since the time from September 1996 to March 1998. Clinical results and CT scanning observation are reported in the following.

  10. 引导种植牙区骨再生的异种脱细胞真皮基质%Acellular dermal matrix used for guiding bone regeneration in the dental implant area

    Institute of Scientific and Technical Information of China (English)

    周静; 邓蔡; 张进锋

    2013-01-01

      BACKGROUND: Acel ular dermal matrix is a kind of prosthodontics membrane material which has been widely used due to the good biocompatibility. OBJECTIVE: To evaluate the effect of acel ular dermal matrix in guiding bone regeneration in the dental implant area. METHODS: Immunohistochemical staining was performed, and the microscope was used to observe the microstructure and cel compatibility of acel ular dermal matrix, in order to evaluate the feasibility of acel ular dermal matrix in guiding bone regeneration. The dental implantation patients who received bone regeneration with acel ular dermal matrix were fol owed-up to evaluate the osteogenesis effect and the effect on soft tissue defects. Then, the effects of Bio-Gide membrane and Bot medical col agen membrane on guiding bone regeneration were compared. RESULTS AND CONCLUSION: The microstructure of acel ular dermal matrix showed there was basement membrane surface and tissue surface. The stylode-like structure and hair fol icle could be observed on the basement membrane surface and the tissue surface was scaly structure, and acel ular dermal matrix had no influence on the proliferation activity osteoblast-like cel s and the alkaline phosphatase activity, but had good cel compatibility. The clinical researches showed that acel ular dermal matrix used in dental implantation was effective for bone regeneration, and there was no significant difference in the effect on guiding bone regeneration when compared with Bio-Gide membrane and Bot medical col agen membrane. The acel ular dermal matrix had good bone regeneration effect in repairing soft tissue deficiencies after bone augmentation.%  背景:异种脱细胞真皮基质属于口腔修复膜材料,因具有良好的生物相容性而被广泛应用。目的:评价异种脱细胞真皮基质在牙区引导牙种植骨再生的效果。方法:以免疫组化染色后显微镜观察异种脱细胞真皮基质的显微结构和细胞相容性,评

  11. 脱细胞软骨基质来源的多孔支架复合山羊髓核细胞体内初步构建组织工程髓核的实验研究%Construction of tissue engineering nucleus pulposus in vivo by combining acellular cartilage matrix derived porous scaffolds with goat nucleus pulposus cells

    Institute of Scientific and Technical Information of China (English)

    伍耀宏; 徐宝山; 杨强; 李秀兰; 张杨; 夏群; 许海委

    2013-01-01

    [Objective] To evaluate the feasibility of construction of tissue engineering nucleus pulposus in vivo by combining acellular cartilage matrix porous scaffolds with PKH26 labeled goat nucleus pulposus cells.[Methods] Porous scaffolds were made of acellular cartilage matrix and evaluated through SEM,Sirius red and HE staining,and toxicity of the scaffolds was assessed by MTT test.P1 generation goat nucleus pulposus cells were identified by safranin O staining and collagen type Ⅱ immunocytochemistry staining.PKH26 labeled cells were seeded onto scaffolds.After 3 d culture in vitro,cell-scaffold hybrids were assessed by LIVE/DEAD staining,then implanted into nude mice subcutaneously for 6w culture.In vivo hybrids were assessed by fluorescence microscope,safranin O staining and collagen type Ⅰ,Ⅱ immunocytochemistry staining.[Results] Pores in scaffold were evenly distributed and connected under SEM,Sirius red and HE staining showed evenly distributed pores.MTT assay demonstrated proliferation among the groups had no significant difference (P > 0.05).P1 generation cells showed chondrocyte-like morphology and stained positively for safranin O and collagen type Ⅱ immunocytochemistry.PKH26 labeled cells showed red fluorescence,cells on scaffolds in vitro showed green fluorescence by LIVE/DEAD staining.After 6w in vivo culture,through fluorescence microscope,safranin O staining and collagen type Ⅰ,Ⅱ immunocytochemistry staining showed positive.[Conclusion] Hybrid of acellular cartilage matrix and goat nucleus pulposus cells can produce nucleus pulposus tissue in vivo.%[目的]探讨脱细胞软骨基质多孔支架复合PKH26标记的山羊髓核细胞体内异位构建组织工程髓核的可行性.[方法]制备脱细胞软骨基质来源的多孔支架,扫描电镜(scanning electron microscope,SEM)观察、天狼星红染色、HE染色观察、MTT毒性检测;分离山羊髓核细胞,通过倒置显微镜观察、番红O染色、Ⅱ型胶原免疫组

  12. 异种脱细胞真皮基质修复膜在口腔黏膜下纤维性变手术治疗中的应用%Clinical application of heterogeneous acellular dermal matrix in the surgical treatment of oral submucous fibrosis

    Institute of Scientific and Technical Information of China (English)

    蒋灿华; 李超; 石芳琼; 陈新群; 唐瞻贵; 翦新春

    2011-01-01

    目的:评价异种脱细胞真皮基质修复膜在口腔黏膜下纤维性变手术治疗中的应用效果.方法:8例重度口腔黏膜下纤维性变患者,经鼻腔气管捕管全麻下切除双侧颊部翼下颌韧带前方区域纵行的纤维条索,术中被动开口度达正常范围后,剪取相应大小的异种脱细胞真皮基质修复膜覆盖黏膜缺损创面,间断缝合后,碘纺纱包加压固定.术后10~14d拆除纱包与缝线后开始开口训练,定期随访并进行类固醇皮质激素黏膜下局部注射等辅助治疗,通过伤口愈合、瘢痕软化及开口度改善等指标评价手术效果.采用SPSS16.0软件包对数据进行单因素方差分析.结果:8例患者双侧颊部纤维条索切除后形成的手术创面,采用异种脱细胞真皮基质修复膜进行修复均获得成功.无感染或排异等并发症发生.术后随访6~18个月,患者颊部原手术区黏膜红润,质地柔软,开口困难明显改善.术前开口度为(12.04±2.93)mm,术中开口度为(35.46±3.17)mm,术后6个月时的开口度为(29.33±4.28)mm,经统计学分析,差异具有显著性(P<0.05).结论:应用异种脱细胞真皮基质修复膜修复重度口腔黏膜下纤维性变手术治疗中的黏膜缺损创面,能够起到促进创面早期愈合、减轻瘢痕形成与改善开口困难的作用,其操作简单易行,值得临床推广应用.%PURPOSE: The aim of this study was to evaluate the clinical effect of heterogeneous acellular dermal matrix in the surgical treatment of advanced oral submucous fibrosis (OSF). METHODS: There were eight patients who had undergone surgical treatment of trismus caused by OSF. Surgery was performed under general anaesthesia given through a nasoendotracheal tube using a fibreoptic bronchoscope. All the fibrous bands on the buccal mucosa were incised and bluntly dissected to stretch the mouth opening. Based on the defect, heterogeneous acellular dermal matrix graft was applied direcdy on the

  13. Astaxanthin reduces matrix metalloproteinase expression in human chondrocytes.

    Science.gov (United States)

    Chen, Wei-Ping; Xiong, Yan; Shi, Yong-Xiang; Hu, Peng-Fei; Bao, Jia-Peng; Wu, Li-Dong

    2014-03-01

    Astaxanthin is a red carotenoid pigment which exerts multiple biological activities. However, little is known about the effects of astaxanthin on matrix metalloproteinases (MMPs) in OA. The present study investigated the effects of astaxanthin on MMPs in human chondrocytes. Human chondrocytes were pretreated with astaxanthin at 1, 10 or 50μM, then, cells were stimulated with IL-1β (10ng/ml) for 24h. MMP-1, MMP-3 and MMP-13 were observed. We found that astaxanthin reduced the expression of MMP-1, MMP-3 and MMP-13 as well as the phosphorylation of two mitogen-activated protein kinases (MAPK) (p38 and ERK1/2) in IL-1β-stimulated chondrocytes. Astaxanthin also blocked the IκB-α degradation. These results suggest that astaxanthin may be beneficial in the treatment of OA.

  14. Guided bone regeneration with acellular dermal matrix as a barrier for bone defects%脱细胞真皮基质膜引导骨缺损成骨变化

    Institute of Scientific and Technical Information of China (English)

    贾仁杰; 任玉卿; 徐昊; 王维英; 弋中萍; 赵保东

    2016-01-01

    BACKGROUND:Acel ular dermal matrix has good biocompatibility and absorbability and exhibits superiority in the guided bone regeneration. OBJECTIVE:To compare the histological changes and osteogenic effects in bone defects after guided bone regeneration with acel ular dermal matrix and Bio-Gide membrane. METHODS:Mandibular second, third and fourth premolars and the first molars bilateral y were extracted from 12 beagle dogs. Three months later, four three-wal bone defect models in the mandible of each dog were made, and randomized into acel ular dermal matrix plus bone graft group (acel ular dermal matrix group), Bio-Gide plus bone graft group (Bio-Gide group), bone graft group, and blank control group (no treatment). In the former two groups, acel ular dermal matrix and Bio-Gide were used to cover the bone grafts, respectively. RESULTS AND CONCLUSION:After surgery, al the beagle dogs recovered wel . Al the groups except the control group showed dramatical improvement in histological changes and percentage of new bone area, and this improvement was more significant in the Bio-Gide and acel ular dermal matrix groups. Moreover, there was no significant difference between the Bio-Gide and acel ular dermal matrix groups. Therefore, the acel ular dermal matrix can be a candidate for bone repair instead of Bio-Gide membrane in the clinical practice.%背景:脱细胞真皮基质膜具有良好的生物相容性、可吸收性、引导骨再生性能。目的:比较脱细胞真皮基质膜和Bio-Gide膜引导骨缺损成骨的组织学变化及引导骨再生的效果的差异。  方法:12只比格犬拔除双侧下颌骨第二、三、四前磨牙及第一磨牙3个月后,在每只犬的下颌骨各建立4处标准的三壁骨缺损模型,随机分为脱细胞真皮基质膜联合骨修复材料组、Bio-Gide膜联合骨修复材料组、骨修复材料组、空白对照组。除空白对照组不做任何处理外,将骨修复材料充实于其余3组骨

  15. Preserving the Posttrapeziectomy Space with a Human Acellular Dermal Matrix Spacer: A Pilot Case Series of Patients with Thumb Carpometacarpal Joint Arthritis

    Directory of Open Access Journals (Sweden)

    Caroline A. Yao, MD

    2013-10-01

    Conclusions: HADM has been used extensively in other forms of reconstruction and has been shown to incorporate into surrounding tissues through neovascularization. Our early results illustrate that HADM can safely fill the dead space left by trapeziectomy.

  16. Evaluation of an extracellular matrix-derived acellular biphasic scaffold/cell construct in the repair of a large articular high-load-bearing osteochondral defect in a canine model

    Institute of Scientific and Technical Information of China (English)

    YANG Qiang; MA Xin-long; HU Yong-cheng; XU Bao-shan; PENG Jiang; LU Shi-bi; GUO Quan-yi; ZHAO Bin; ZHANG Li; WANG Ai-yuan; XU Weng-jing; XIA Qun

    2011-01-01

    Background Osteochondral lesion repair is a challenging area of orthopedic surgery.Here we aimed to develop an extraceliular matrix-derived,integrated,biphasic scaffold and to investigate the regeneration potential of the scaffold loaded with chondrogenically-induced bone marrow-derived mesenchymal stem cells (BMSCs) in the repair of a large,high-load-bearing,osteochondral defect in a canine model.Methods The biphasic scaffolds were fabricated by combining a decellularization procedure with a freeze-drying technique and characterized by scanning electron microscopy (SEM) and micro-computed tomography (micro-CT).Osteochondral constructs were fabricated in vitro using chondrogenically-induced BMSCs and a biphasic scaffold,then assessed by SEM for cell attachment.Osteochondral defects (4.2 mm (diameter) x 6 mm (depth)) were created in canine femoral condyles and treated with a construct of the biphasic scaffold/chondrogenically-induced BMSCs or with a cell-free scaffold (control group).The repaired defects were evaluated for gross morphology and by histological,biochemical,biomechanical and micro-CT analyses at 3 and 6 months post-implantation.Results The osteochondral defects of the experimental group showed better repair than those of the control group.Statistical analysis demonstrated that the macroscopic and histologic grading scores of the experimental group were always higher than those of the control group,and that the scores for the experimental group at 6 months were significantly higher than those at 3 months.The cartilage stiffness in the experimental group (6 months) was (6.95±0.79)N/mm,70.77% of normal cartilage; osteochondral bone stiffness in the experimental group was (158.16±94.30) N/mm,74.95% of normal tissue; glycosaminoglycan content of tissue-engineered neocartilage was (218±21.6) μg/mg (dry weight),84.82% of native cartilage.Micro-CT analysis of the subchondral bone showed mature trabecular bone regularly formed at 3 and 6 months

  17. Exogenic Acellular Dermal Matrix in Guided Bone Regeneration of Dental Implant%异种脱细胞真皮基质在牙种植中引导骨再生的临床观察

    Institute of Scientific and Technical Information of China (English)

    韦丽萍; 左陈启; 王远勤

    2011-01-01

    Objective: To identify the clinical effect of a homemade exogenic acelluar dermal matrix as a barrier membrane in guide bone regeneration (GBR) of dental implants. Methods: Seventy eight dental implants/cases with limited bone-bed were divided into 2 groups, and treated by GBR technology with different barrier membrane. Experiment group (38 cases) used homemade exogenic acelluar dermal matrix, while in the control group (40 cases) Bio-Gide biofilm was used. Results: The difference of bone harvested in two groups was not statistically significant (P>0.05). All patients were healed with first intention and the healing rate of 2 groups were all 100%. One case complicated with facial swelling in control group, which was cured after anti-inflammatory and symptomatic treatment. There was no statistically significant between two groups in the demographic basic information, such as, adverse events, healing of incision, bone growth effect,and the like. Conclusion: This alternative homemade material should be considered in GBR by practitioners.%目的:观察国产异种脱细胞真皮基质修复膜作为屏障膜,在牙种植的引导骨再生中应用的临床效果.方法:78例需要引导骨再生的种植病例,均为前牙区或前磨牙区单牙或连续多牙位(3牙)缺失;其中前牙52颗,前磨牙26颗.缺牙区牙槽嵴主要为唇颊侧垂直性吸收,牙槽嵴顶宽度约1~3 mm,牙槽嵴水平吸收量在2 mm以内.分为2组,一组采用国产异种脱细胞真皮基质修复膜作为GBR技术的屏蔽膜(38例),另一组采用Bio-Gide生物膜作为对照(40例),比较二者的临床效果.结果:2组在骨生长效果之间的差异无统计学意义(P>0.05).2组患者切口均甲级愈合,切口愈合率均为100%.术后3 d,对照组有1例发生面部肿胀,经抗感染治疗和对症处理后缓解.本研究的其他病例均未出现感染等不良事件.结论:使用国产异种脱细胞真皮基质修复膜在牙种植术中进行骨引

  18. Human keratinocytes synthesize and secrete the extracellular matrix protein, thrombospondin.

    Science.gov (United States)

    Wikner, N E; Dixit, V M; Frazier, W A; Clark, R A

    1987-02-01

    Thrombospondin (TSP) a glycoprotein originally identified as the endogenous lectin of platelets, is also synthesized by fibroblasts, endothelial cells, pneumocytes, smooth muscle cells, and macrophages. Thrombospondin is subdivided into functional domains which bind specifically to heparin, fibronectin, collagen, and to specific cellular receptors. It is found within the basement membranes of kidney, lung, smooth muscle, and skin. Thus TSP may serve as an important link between cells and matrices. Thrombospondin also has been reported at the epidermal-dermal junction. We wished to determine whether human keratinocytes synthesize and secrete TSP. Pure human keratinocytes were grown in defined medium without fibroblast feeder layers. Immunofluorescent staining with either rabbit polyclonal or mouse monoclonal antibodies to human platelet TSP yielded specific granular staining within the cytoplasm of keratinocytes. Culture media and cellular lysates were harvested from cultures metabolically labeled with [35S]methionine. Trichloroacetic acid precipitation, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and autoradiography revealed a major labeled band comigrating with purified platelet TSP in both the media and the cellular lysates. Immunoprecipitation with either the polyclonal or the monoclonal anti-TSP antibodies followed by SDS-PAGE and autoradiography identified this band as TSP. Thus keratinocytes in culture synthesize and secrete TSP. Thrombospondin may play an important role in epidermal interactions with extracellular matrix.

  19. Extracellular Matrix Molecular Remodeling in Human Liver Fibrosis Evolution.

    Directory of Open Access Journals (Sweden)

    Andrea Baiocchini

    Full Text Available Chronic liver damage leads to pathological accumulation of ECM proteins (liver fibrosis. Comprehensive characterization of the human ECM molecular composition is essential for gaining insights into the mechanisms of liver disease. To date, studies of ECM remodeling in human liver diseases have been hampered by the unavailability of purified ECM. Here, we developed a decellularization method to purify ECM scaffolds from human liver tissues. Histological and electron microscopy analyses demonstrated that the ECM scaffolds, devoid of plasma and cellular components, preserved the three-dimensional ECM structure and zonal distribution of ECM components. This method has been then applied on 57 liver biopsies of HCV-infected patients at different stages of liver fibrosis according to METAVIR classification. Label-free nLC-MS/MS proteomics and computation biology were performed to analyze the ECM molecular composition in liver fibrosis progression, thus unveiling protein expression signatures specific for the HCV-related liver fibrotic stages. In particular, the ECM molecular composition of liver fibrosis was found to involve dynamic changes in matrix stiffness, flexibility and density related to the dysregulation of predominant collagen, elastic fibers and minor components with both structural and signaling properties. This study contributes to the understanding of the molecular bases underlying ECM remodeling in liver fibrosis and suggests new molecular targets for fibrolytic strategies.

  20. Application of acellular dermal matrix and expandedflapin half auricular reconstructionwithrib cartilage grafts%脱细胞真皮及扩张皮瓣在肋软骨移植半耳缺损再造中的应用

    Institute of Scientific and Technical Information of China (English)

    董海江; 王喜梅; 万程; 李想; 张琼阁

    2016-01-01

    BACKGROUND:Traumatic auricle defectsin upper 1/2 or lower 1/2,seriously involve theauricular cartilage and skin blood vessels. The autogenic rib cartilage graft and acelular dermal matrix have good histocompatibility, and expanded flapis a kind of thin and achromatic tissue for skin defect repair. OBJECTIVE:To explore theapplication ofacelular dermal matrix and expanded flap in half auricular reconstruction,and to find out the fine carving and anastomosis of autogenic rib cartilage graftas wel as its similarities with the ear and clinical significance. METHODS:Eight cases of half auricular defects were treated with expanded flap, autogenic rib cartilage graft, fine anastomosisofautogenic rib cartilage graft and residual earfor half auricular reconstruction,during which theacelular dermal matrixwas usedto promote residual ear docking and skul auricle angle formation. The reconstructionwasperformed in three stages:first,anexpander(volume, 80mL)wassubcutaneously implanted attheretro-auricular area;second, the auricular defects were reconstructed with fine rib cartilage graft, acelular dermal matrix and auriculoplasty;finaly, acelular dermal matrixwas usedto promote residual ear docking. Thenthehalf auricular reconstructionwas evaluatedby objective measurement and subjective rating. RESULTS AND CONCLUSION:Half auricular reconstruction was successful in al the eight caseswithout obvious complications, and the cartilage grafts were in good condition.During thefolow-up,thereconstructed auriclewasshapedwel andformed a good involution withtheresidualauricle. In addition, the flange was smooth withoutobviouscolor difference and edema, and its position, size and shape were consistent with those of thecontralateralone. Afterthefolow-up of 6 months, objective indicators showed that the affected side had no significant differencefromthe contralateralone(P> 0.05). In conclusion,theacelular dermal matrixcanobviously decrease the complications of the cartilage grafts

  1. Acellularization-Induced Changes in Tensile Properties Are Organ Specific - An In-Vitro Mechanical and Structural Analysis of Porcine Soft Tissues.

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    Stefan Schleifenbaum

    Full Text Available Though xenogeneic acellular scaffolds are frequently used for surgical reconstruction, knowledge of their mechanical properties is lacking. This study compared the mechanical, histological and ultrastructural properties of various native and acellular specimens.Porcine esophagi, ureters and skin were tested mechanically in a native or acellular condition, focusing on the elastic modulus, ultimate tensile stress and maximum strain. The testing protocol for soft tissues was standardized, including the adaption of the tissue's water content and partial plastination to minimize material slippage as well as templates for normed sample dimensions and precise cross-section measurements. The native and acellular tissues were compared at the microscopic and ultrastructural level with a focus on type I collagens.Increased elastic modulus and ultimate tensile stress values were quantified in acellular esophagi and ureters compared to the native condition. In contrast, these values were strongly decreased in the skin after acellularization. Acellularization-related decreases in maximum strain were found in all tissues. Type I collagens were well-preserved in these samples; however, clotting and a loss of cross-linking type I collagens was observed ultrastructurally. Elastins and fibronectins were preserved in the esophagi and ureters. A loss of the epidermal layer and decreased fibronectin content was present in the skin.Acellularization induces changes in the tensile properties of soft tissues. Some of these changes appear to be organ specific. Loss of cross-linking type I collagen may indicate increased mechanical strength due to decreasing transverse forces acting upon the scaffolds, whereas fibronectin loss may be related to decreased load-bearing capacity. Potentially, the alterations in tissue mechanics are linked to organ function and to the interplay of cells and the extracellular matrix, which is different in hollow organs when compared to skin.

  2. Acellular Dermal Matrix as GTR Barrier Membrane on Periodontal Regeneration in the Treatment of Class Ⅱ Furcation Defects%脱细胞真皮基质作为GTR屏障膜治疗Ⅱ度根分叉缺损的实验研究

    Institute of Scientific and Technical Information of China (English)

    王乾锋; 刘宏伟

    2013-01-01

    目的:观察脱细胞真皮基质(acellular dermal matrix,ADM) 作为引导组织再生(guided tissue regeneration,GTR)屏障膜在治疗Ⅱ度根分叉缺损时的牙周组织再生情况.方法:在犬的两侧下颌第三、四前磨牙制造Ⅱ度根分叉缺损模型,将ADM作为GTR屏障膜覆盖在根分叉缺损区表面,于术后8周观察和测量根分叉处牙周组织的再生情况,并与空白对照组作比较.结果:术后8周,ADM组和空白对照组的临床附着丧失(clinical attachment loss,CAL)平均分别为1.90 mm 和2.85 mm,差异有统计学意义(P<0.05);ADM 组的新骨面积、新骨高度、新生牙骨质高度分别为8.23 mm2、4.52 mm、4.72 mm,明显大于对照组的1.75 mm2、0.91 mm、0.94 mm,而上皮和结缔组织面积则小于对照组,分别为0.02、0.54 mm2 和0.10、5.56 mm2,差异均有统计学意义(P<0.05).结论:ADM 作为GTR屏障膜治疗下颌Ⅱ度根分叉缺损,能比空白对照组获得更多的临床附着和再生牙周组织.

  3. Effects of hyaluronic acid on biomechanic performance of porcine acellular dermal matrix plus thin skin autograft after transplantation%透明质酸对复合移植皮肤组织生物力学性能影响的实验观察

    Institute of Scientific and Technical Information of China (English)

    赵京玉; 柴家科; 宋慧锋; 许明火

    2012-01-01

    Objective To explore the effects of hyaluronic acid (HA) on biomechanical properties for porcine acellular dermal matrix (PADM) plus thin skin autograft after transplantation.Methods The dorsa of 10 Japanese white rabbits were symmetrically divided into four areas of A-D by random grouping.Full-thickness skin defects were created in Groups A-C while Group D was blank with normal skin.Operations were performed in Group A:implant with HA + PADM + thin skin autografts,Group B:implant with PADM + thin skin autografts and Group C:skin autografts group.Histological examination of specimen was performed at Day 56 postoperatively.And the biomechanical properties such as relaxation and stress-strain properties of grafts were recorded.Results The structure of PADM was found to be basically intact by hematoxylin and eosin E dyeing in Groups A and B.In Group A,dense fiber structure could be observed.Lots of regularly arranged collagenous fibers and new blood capillaries were grown into the dermal matrix with sparsely distributed inflammatory cells.In Group B,acellular dermal matrix became clustered with a small amount of invaded fibroblasts.And there was a high expression of inflammatory cells.The biomechanic pedormances of transplanted skin were:Group A's curve was mostly close to that of Group D's,Group B's curve was the most further from that of Group D's (P =0.001 ) and Group G's curve stayed between Groups A and B.Under the same strain,the stress of Groups A-D was ( 87 ± 8 ),( 115 ± 9 ),(60 ± 7 ) and (81 ± 4) kPa respectively.No significant difference of stress existed between these two groups (P=0.838).There was significant difference of stress between Groups B/C and D (P =0.001 and P =0.009).Conclusion Topical hyaluronic acid may be used to enhance the biomechanics pedormances of transplanted skin.%目的 探讨外用透明质酸(HA)对异种(猪)去细胞真皮基质( PADM)复合移植皮肤组织生物力学性能的影响.方法 选用10只日本大耳兔作为

  4. Repair of radial and digital nerve defect with human acellular nerve allograft:6 cases report%去细胞同种异体神经移植修复桡神经和指神经缺损六例

    Institute of Scientific and Technical Information of China (English)

    唐举玉; 俞芳; 吴攀峰; 黄臻; 梁捷予; 何波; 刘小林

    2014-01-01

    Objective To explore the safety and clinical effect of the human acellular nerve allograft (hANG) for repairing peripheral nerve defects.Methods During November,2009 to October,2010,6 patients with 3 digital nerve defects and 3 radial nerve defects were repaired with hANG.During postoperation period,safety was evaluated by local wound response and laboratory testing,while the efficacy was evaluated by British Medical Research Council sensory function assessment standards,static 2-point discrimination (2PD) and Semmes-Weinstein monofilament testing.Results Three patients with 6 digital nerve defects received hANG transplant.The length of nerve graft was 20-50 mm(mean 30.8 mm).After followed up for 31-40 months,the excellent rate of 2PD was 66.7%.Two of 3 patients rahabilited as well as the normal.Three patients with radial nerve defects,whose length of nerve graft was 35-60 mm(mean 48.3 mm).The strength of extensor carpiradialis longus muscle had restored Ⅲ in 1 case,and other 2 cases had no restoration.Conclusion hANG is safe and effective for repairing peripheral nerve defects,especially for digital nerve defects.%目的 探讨去细胞同种异体神经(hANG)移植修复周围神经缺损的安全性和有效性. 方法 2009年9月-2010年10月,应用hANG移植修复周围神经缺损6例,其中指神经缺损3例、桡神经缺损3例,术后观察伤口愈合情况及生化、免疫学检查,采用英国医学研究会感觉功能评定标准、Semmes-Weinstein单丝触觉和静态两点辨别觉(2PD)评价hANG的临床效果. 结果 所有病例切口术后无红肿及渗出、愈合良好.3例指神经损伤患者共有6条指神经缺损,神经移植长度20~ 50 mm(平均30.8 mm),随访31~40个月,静态2PD优良率66.7%,其中2例4条指神经缺损患者术后感觉基本恢复正常;3例桡神经损伤患者,神经移植长度35 ~ 60 mm(平均48.3 mm),随访18 ~ 36个月,其中l例桡侧腕伸肌肌力恢复至Ⅲ级,术后

  5. Use of a collagen-elastin matrix as transport carrier system to transfer proliferating epidermal cells to human dermis in vitro.

    Science.gov (United States)

    Waaijman, Taco; Breetveld, Melanie; Ulrich, Magda; Middelkoop, Esther; Scheper, Rik J; Gibbs, Susan

    2010-01-01

    This in vitro study describes a novel cell culture, transport, and transfer protocol that may be highly suitable for delivering cultured proliferating keratinocytes and melanocytes to large open skin wounds (e.g., burns). We have taken into account previous limitations identified using other keratinocyte transfer techniques, such as regulatory issues, stability of keratinocytes during transport (single cell suspensions undergo terminal differentiation), ease of handling during application, and the degree of epidermal blistering resulting after transplantation (both related to transplanting keratinocyte sheets). Large numbers of proliferating epidermal cells (EC) (keratinocytes and melanocytes) were generated within 10-14 days and seeded onto a three-dimensional matrix composed of elastin and collagen types I, III, and V (Matriderm®), which enabled easy and stable transport of the EC for up to 24 h under ambient conditions. All culture conditions were in accordance with the regulations set by the Dutch Central Committee on Research Involving Human Subjects (CCMO). As an in vitro model system for clinical in vivo transfer, the EC were then transferred from Matriderm onto human acellular dermis during a period of 3 days. After transfer the EC maintained the ability to regenerate into a fully differentiated epidermis containing melanocytes on the human dermis. Proliferating keratinocytes were located in the basal layer and keratin-10 expression was located in differentiating suprabasal layers similar to that found in human epidermis. No blistering was observed (separation of the epidermis from the basement membrane). Keratin-6 expression was strongly upregulated in the regenerating epidermis similar to normal wound healing. In summary, we show that EC-Matriderm contains viable, metabolically active keratinocytes and melanocytes cultured in a manner that permits easy transportation and contains epidermal cells with the potential to form a pigmented reconstructed

  6. Central role of pyrophosphate in acellular cementum formation.

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    Brian L Foster

    Full Text Available BACKGROUND: Inorganic pyrophosphate (PP(i is a physiologic inhibitor of hydroxyapatite mineral precipitation involved in regulating mineralized tissue development and pathologic calcification. Local levels of PP(i are controlled by antagonistic functions of factors that decrease PP(i and promote mineralization (tissue-nonspecific alkaline phosphatase, Alpl/TNAP, and those that increase local PP(i and restrict mineralization (progressive ankylosis protein, ANK; ectonucleotide pyrophosphatase phosphodiesterase-1, NPP1. The cementum enveloping the tooth root is essential for tooth function by providing attachment to the surrounding bone via the nonmineralized periodontal ligament. At present, the developmental regulation of cementum remains poorly understood, hampering efforts for regeneration. To elucidate the role of PP(i in cementum formation, we analyzed root development in knock-out ((-/- mice featuring PP(i dysregulation. RESULTS: Excess PP(i in the Alpl(-/- mouse inhibited cementum formation, causing root detachment consistent with premature tooth loss in the human condition hypophosphatasia, though cementoblast phenotype was unperturbed. Deficient PP(i in both Ank and Enpp1(-/- mice significantly increased cementum apposition and overall thickness more than 12-fold vs. controls, while dentin and cellular cementum were unaltered. Though PP(i regulators are widely expressed, cementoblasts selectively expressed greater ANK and NPP1 along the root surface, and dramatically increased ANK or NPP1 in models of reduced PP(i output, in compensatory fashion. In vitro mechanistic studies confirmed that under low PP(i mineralizing conditions, cementoblasts increased Ank (5-fold and Enpp1 (20-fold, while increasing PP(i inhibited mineralization and associated increases in Ank and Enpp1 mRNA. CONCLUSIONS: Results from these studies demonstrate a novel developmental regulation of acellular cementum, wherein cementoblasts tune cementogenesis by modulating

  7. Comparison of autogenous cartilage, acellular dermis, and solvent-dehydrated pericardium for the prevention and correction of dorsal nasal irregularities: an experimental study.

    Science.gov (United States)

    Çöloğlu, Harun; Uysal, Afşin; Tiftikçioğlu, Yiğit Özer; Oruç, Melike; Koçer, Uğur; Coşkun, Erhan; Ramadan, Selma Uysal; Astarcı, Müzeyyen Hesna

    2012-06-01

    Numerous materials have been used for the correction and prevention of dorsal nasal irregularities. Experimental and clinical studies have been useful but have provided insufficient results for several reasons, including the impossibility of obtaining pathologic specimens from aesthetic patients and imprecise experimental models. In this study, an experimental model for rhinoplasty is used for the comparative evaluation of solvent-dehydrated pericardium, acellular dermal matrix, and autogenous ear cartilage as onlay grafts for the prevention and correction of nasal dorsal irregularities. We used an experimental rabbit rhinoplasty model that has a human nose-like osteocartilaginous junction. Thus, our goal is to get a more realistic idea about the features of these three materials. Thirty New Zealand rabbits weighing 2,100-2,550 g were used. The noses of the rabbits were evaluated with computerized tomographic measurements, "pinch" tests were performed for skin properties, and all were photographed before the surgical procedures. They were divided into three groups: Autogenous cartilage grafts were applied after the rhinoplasty operation in group 1, acellular dermal matrixes were used after the rhinoplasty in group 2, and pericardium allografts were used after the rhinoplasty in group 3. The rabbits were followed up for 4 months before they were evaluated by photography, computerized tomography, and "pinch" tests for the skin properties of the nose. Then they were killed for histopathologic evaluation. Adhesion and resorption rates of the onlay grafts were observed and subdermal thickness measurements were made to determine the fate of the grafts as well as their effects on the overlying skin. The major advantages of the allografts used in groups 2 and 3 are the ease of obtaining them without any donor site morbidity, shorter operative procedures, and lower distortion rates due to lack of cartilage memory. The results of this study conform to those of previous

  8. Effect of schedule on reactogenicity and antibody persistence of acellular and whole-cell pertussis vaccines: value of laboratory tests as predictors of clinical performance.

    Science.gov (United States)

    Miller, E; Ashworth, L A; Redhead, K; Thornton, C; Waight, P A; Coleman, T

    1997-01-01

    The performance of four acellular pertussis vaccines containing between two and five pertussis antigens combined with diphtheria and tetanus toxoids was compared with that of British whole-cell diphtheria/tetanus/pertussis (DTP) vaccine both in laboratory assays for potency, toxicity and immunogenicity, and for reactogenicity and immunogenicity in infants. Clinical responses were evaluated in double blind randomized Phase II trials using 3/5/9 month and 2/3/4 month schedules. The acellular DTPs had much lower toxicity than whole-cell DTP in laboratory tests and were significantly less pyrogenic than whole-cell DTP under both schedules. Local reactions were not consistently lower in acellular than whole-cell vaccinees and varied with the source of the diphtheria and tetanus antigens used. Differences in endotoxin level and content of active pertussis toxin (PT) between acellular DTP vaccines were not clinically significant. The reactogenicity advantage of the acellular vaccines was substantially reduced under the 2/3/4 month schedule due to the reduced reactogenicity of the whole-cell DTP vaccine when given at a younger age. There was no relationship between antigen content measured in micrograms per dose and ELISA antibody responses to filamentous haemagglutinin (FHA) and PT in infants, nor was murine immunogenicity predictive of immunogenicity in humans. Antibody response to PT was attenuated in the whole-cell group under the 2/3/4 month schedule but was unaffected in the group receiving acellular vaccines with individually purified components; antibody response to pertactin (69 kDa antigen) was similar in recipients of the whole-cell and component acellular vaccines under the 2/3/4 month schedule. PT antibody persistence until 4-5 years of age was significantly better in recipients of the component acellular than either the whole-cell vaccine or the co-purified acellular vaccine under the 3/5/9 month schedule. However, diphtheria antitoxin levels were reduced in

  9. Long-Term Followup of Dermal Substitution with Acellular Dermal Implant in Burns and Postburn Scar Corrections

    OpenAIRE

    Juhasz, I.; Kiss, B.; Lukacs, L.; Erdei, I.; Peter, Z.; Remenyik, E.

    2010-01-01

    Full-thickness burn and other types of deep skin loss will result in scar formation. For at least partial replacement of the lost dermal layer, there are several options to use biotechnologically derived extracellular matrix components or tissue scaffolds of cadaver skin origin. In a survey, we have collected data on 18 pts who have previously received acellular dermal implant Alloderm. The age of these patients at the injury varied between 16 months and 84 years. The average area of the impl...

  10. Meshed acellular dermal matrix:technique and application in implant based breast reconstruction

    Institute of Scientific and Technical Information of China (English)

    Dino Zammit; Jonathan Kanevsky; Fan-Yi Meng; Tassos Dionisopoulos

    2016-01-01

    Alloderm was the first acellular dermal matrix used and remains a popular choice among plastic surgeons. However, while the overall surgical outcome of breast reconstruction using alloderm has been a success, the economic burden on the health care system makes it a subject of frequent re-evaluations in cost-effectiveness. Prompted by the high price of $3,700 USD for a 6 cm × 16 cm area, our group proposes the meshing of AlloDerm to decrease the total amount needed for breast reconstruction, while achieving comparable surgical outcomes as using unmeshed alloderm.

  11. Immunohistochemical Studies of the Expression of Matrix Metalloproteinase-2 and Metalloproteinase-9 in Human Prostate Cancer

    Institute of Scientific and Technical Information of China (English)

    曾汉青; 肖亚军; 鲁功成; 陈勇

    2003-01-01

    To study the expression of matrix metalloproteinase-2 and -9 in human prostate cancer,matrix metalloproteinase-2 and -9 were immunohistochemically detected in tissues of prostate cancer and benign prostatic hyperplasia (BPH). Our results showed that matrix metalloproteinase-2 and -9 levels in prostate cancer were much higher than those in tissues of BPH, with the cancer invasion being positively correlated with the expression of the metalloproteinases. It is concluded that matrix metalloproteinase-2 and -9 are better molecular markers, which are of help in the diagnosis and prediction of prognosis of prostate cancer.

  12. Plasma matrix metalloproteinase-9 response to downhill running in humans.

    Science.gov (United States)

    Welsh, M C; Allen, D L; Byrnes, W C

    2014-05-01

    Matrix metalloproteinase-9 is a proteolytic enzyme capable of degrading proteins of the muscle extracellular matrix. Systemic levels of MMP-9 or its inhibitor, tissue inhibitor of metalloproteinase-1 (TIMP-1), have the potential to serve as blood markers of exercise-induced muscle damage. The purpose of this study was to determine if an eccentrically-dominated task, downhill running (DHR), produces changes in plasma MMP-9 or TIMP-1 and examine the relationship between MMP-9/TIMP-1 levels and indirect indicators of muscle damage. Subjects were sedentary (SED, n=12) or had a history of concentrically-biased training (CON, n=9). MMP-9 and TIMP-1 were measured before (Pre-Ex), immediately after (Post-Ex), and 1-, 2-, 4-, and 7-days post-DHR (-10°), and compared to discomfort ratings, creatine kinase activity and strength loss. At 1-day Post-Ex, discomfort increased (5.6 ± 7.8 to 45.5 ± 19.9 mm; 0-100 mm scale), strength decreased (-6.9 ± 1.6%) and CK increased (162.9 ± 177.2%). MMP-9 was modestly but significantly increased at Post-Ex in both CONC and SED (32.7 ± 33.6%) and at 4-days in SED (66.9 ± 88.1%), Individual responses were variable, however. There were no correlations between MMPs and discomfort ratings, plasma CK or strength. While plasma MMP-9 changes may be detectable in the systemic circulation after DHR, they are small and do not correspond to other markers of damage. © Georg Thieme Verlag KG Stuttgart · New York.

  13. Effectiveness of Acellular Dermal Matrix Prophylaxis in Mandibular Impacted Molars Extraction :A Meta-analysis%脱细胞真皮基质预防下颌阻生磨牙拔除术后并发症效果的Meta分析

    Institute of Scientific and Technical Information of China (English)

    解龙川; 徐晓明; 曾宪涛

    2012-01-01

    Objective To evaluate the efficacy of acellular dermal matrix (ADM) prophylaxis in mandibular impacted molars (IM) extraction by performing a meta-analysis. Methods The PubMed, CENTRAL, SinoMed.CNKI, VIP, and Wanfang databases were searched from 1999 to July 2012,in order to retrieve relevant studies. Manual searching was also performed. After assessing the methodological quality and data extraction, a meta-analysis was conducted by using the Meta-Analyst 3. 13 software;and the levels of evidences were assessed according to GRADE system by using GRADEpro 3. 6 software. Results Twelve RCTs and CCTs involing 1 267 teeth(ADM group) were included. The meta-analysis showed that compared with blank control group,the ADM could obviously decrease the rate of dry socket(RR=0.14,95%CI =0.09 ~0.24,P< 0. 001) ,rebleeding(RR = 0. 34,95% CI =0. 20 ~ 0. 60,P < 0. 001) ,swelling( RR = 0. 58,95% CI =0. 35 ~ 0. 96,P = 0.04) ,and trismus( RR =0.72,95%CI =0.52~0.99,P =0.04) ;in sequence,the levels of evidences were high,morder-ate,very low,and low.Conclusions Based on current evidences,ADM can effectively reduce the complications after mandibular IM extraction.However,large sample,randomised,controlled,as an index of 'cost-benefit analysis' ,and according to CONSORT statement trails are suggested,before recommending ADM as a routine protocol.%目的:采用Meta分析的方法综合评价脱细胞真皮基质(ADM)预防下颌阻生磨牙拔除术后并发症的效果.方法:计算机检索PubMed、CENTRAL、SinoMed、CNKI、VIP和万方数据库中1999年至2012年7月发表的相关研究,并辅以手检.对符合纳入标准的研究进行质量评价和资料提取后,采用Meta-Analyst 3.13软件进行Meta分析,并遵照GRADE系统采用GRADEpr0 3.6软件进行证据等级评定.结果:共纳入12个(半)随机临床对照试验,共1267例使用ADM的患者.Meta分析结果表明,与空白对照相比,脱细胞真皮基质可以显著降低86%发生干槽症的风险(RR =0.14,95

  14. 脱细胞真皮基质用于前牙即刻种植扩增角化龈的临床研究%Clinical Observation of Peri-implants Keratinized Tissue Augmented in Anterior Teeth with Acellular Dermal Matrix Xerograft

    Institute of Scientific and Technical Information of China (English)

    彭国光; 谢建雅; 夏炜; 严鑫; 吴俊伟; 沈琳; 吴美珍; 谭玉莲

    2011-01-01

    Objective To evaluate the effectiveness of acellular dermal matrix (ADM) xerograft in increasing the width and esthetics of keratinized gingiva around anterior immediate dental implants. Methods Twenty patients received ADM xerograft with/without guided bone regeneration ( GBR ) after immediate implants in maxillary or mandibular anterior regions. The width of keratinized gingiva was recorded at 3 and 6 months after final porcelain crown restoration. The parameter was compared to the mean width of adjacent teeth, and the implant gingival papilla was evaluated with Jemt' s classification. Results ADM xerografts provided satisfactory results. The width and esthetics of the gingival/papilla in immediate implants were no difference (P >0.05) with them in the natural adjacent teeth, and the position before operation. Conclusion The width of keratinized tissues was augmentation by using the ADM xerograft.%目的 研究脱细胞真皮基质用于前牙即刻种植扩增角化龈的临床效果.方法 唇侧骨板垂直缺损不超过牙根长度1/3的单颗前牙即刻种植病例20例,拔除患牙即刻种植,利用异种脱细胞真皮基质双层封闭植牙创口,并与周围黏膜加压严密缝合,2~3周拆线,8~12周行冠修复.冠修复后3、6个月,从龈缘高点到膜龈联合线测量种植牙角化龈的宽度,和邻牙及术前角化龈的宽度进行比较,并对种植牙的龈乳头进行美学评价.结果 20颗种植牙的角化龈宽度与种植前相比无明显差异,与相邻牙也无明显差异,膜龈联合线自然;17例种植牙的龈乳头达到Jemt氏分类的2级、3级.种植前,20颗种植牙的角化龈宽度为(4.460±0.220) mm,冠修复后3个月为(4.451±0.245) mm,正常邻牙是(4.410±0.189) mm.冠修复后3个月,种植位点角化龈平均宽度与正常邻牙比较(t =1.283,P=0.215)、与术前比较(=0.584,P=0.566),差异均无统计学意义.冠修复后6个月,种植位点角化龈宽度为(4.448±0.223) mm

  15. Pulmonary heart valve replacement using stabilized acellular xenogeneic scaffolds; effects of seeding with autologous stem cells

    Directory of Open Access Journals (Sweden)

    Harpa Marius Mihai

    2015-12-01

    Full Text Available Background: We hypothesized that an ideal heart valve replacement would be acellular valve root scaffolds seeded with autologous stem cells. To test this hypothesis, we prepared porcine acellular pulmonary valves, seeded them with autologous adipose derived stem cells (ADSCs and implanted them in sheep and compared them to acellular valves.

  16. The Use of the Matrix Method for the Study of Human Motion:Theory and Applications

    Institute of Scientific and Technical Information of China (English)

    Zong-Ming Li; Jesse A. Fisk; Savio L-Y. Woo

    2003-01-01

    Kinematics has been successfully used to describe body motion without reference to the kinetics (or forces causing the motion). In this article, both the theory and applications of the matrix method are provided to describe complex human motion. After the definition of a Cartesian coordinate frame is introduced, the description of transformations between multiple coordinate frames is given; the decomposition of a transformation matrix into anatomical joint motion parameters (e.g. Euler angles) is then explained. The advantages of the matrix method are illustrated by three examples related to biomechanical studies. The first describes a reaching and grasping task in which matrix transformations are applied to position the hand with respect to an object during grasping. The second example demonstrates the utility of the matrix method in revealing the coupling motion of the wrist between flexion-extension and radial-ulnar deviation. The last example highlights the indispensable use of the matrix method for the study of knee biomechanics, including the description of knee joint kinematics during functional activities and determination of in-situ ligament forces using robotic technology, which has advanced our understanding of the functions of the cruciate ligaments to knee joint kinematics. It is hoped that the theoretical development and biomechanical application examples will help the readers apply the matrix method to research problems related to human motion.

  17. Comparison of recombinant human granulocyte - macrophage colony stimulating factor gel and acellular skin of treating deep second degree burn wound in clinical effect%重组人粒细胞巨噬细胞集落刺激因子凝胶与脱细胞异种皮治疗深Ⅱ度烧伤创面的临床效果比较

    Institute of Scientific and Technical Information of China (English)

    张留栓; 田彭

    2016-01-01

    Objective To investigate the effects of recombinant human granulocyte macrophage colony stimulating factor gel and the clini-cal effect of acellular xenogeneic skin for treating deep second degree burn wounds. Methods Between 2010 January 2013 to January in our hos-pital,deep second degree burn patients in 60 cases,using a random number table method patients were divided into three groups and were recor-ded as a group treated with rhGM CSF therapy),group B(given off acellular skin treatment)and group C treated with traditional methods of treat-ment),20 cases in each group. The 7,14,21 days of wound healing,7 days of wound bacteria detection rate,scar score were observed. Results A,B two groups compared to the C group,wound healing rate of 7,14,21 days is improved significantly,healing time significantly is lower, and the difference is statistically significant( P 0. 05). Between a group and C group, the bacterial detection rate at 7 days significantly decreased( P 0.05);而与 C 组对比,A 组7 d 细菌检出率有显著性的降低( P <0.01)。A、B 两组在色泽、柔软度、厚度、血管分布等4个方面较 C 组显著降低(均 P <0.01);与 B 组比较,A 组在柔软度、厚度、血管分布等4个方面也有显著性的降低(均 P <0.05)。结论在深Ⅱ度烧伤的创伤修复早期阶段,rhGM - CSF 凝胶与脱细胞异种皮治疗都具有良好的治愈率,各具有其不同的优势。

  18. Activity of MMP-9 after repair of abdominal wall defects with acellular and crosslinked bovine pericardium in rabbit.

    Science.gov (United States)

    Singh, Himani; Kumar, Naveen; Sharma, A K; Kataria, Meena; Munjal, Ashok; Kumar, Amit; Dewangan, Rukmani; Kumar, Vineet; Devarathnam, J; Kumar, Sachin

    2014-02-01

    This study was undertaken for the identification of matrix metalloproteinases (MMPs) in extracts obtained from native, acellular and crosslinked bovine pericardium (in vitro), as well as in the plasma after implantation of these biomaterials in rabbits (in vivo). Native pericardium (NP) expressed a 72 kDa (MMP-2) band; whereas, in acellular pericardium (AP) two bands (10 kDa and 92 kDa) of MMPs were observed of which, 92 kDa band was very faint. AP crosslinked with glutaraldehyde did not show any gelatinase activity and thus reflects the creation of new additional chemical bonds between the collagen molecules which has been effectively removed. Gelatin zymography showed only one major band of 92 kDa in all the implanted and untreated rabbit plasma, but the relative amount of 92 kDa was 1-2 times higher in acellular bovine pericardium implanted rabbits as compared to crosslinked and native groups. In NP group, the 92 kDa band was the dullest among the three groups. This indicated that the level of MMP-9 corresponds to the degree of collagen degradation. © 2012 The Authors. International Wound Journal © 2012 Medicalhelplines.com Inc and John Wiley & Sons Ltd.

  19. Sterile Acellular Dermal Collagen as a Treatment for Rippling Deformity of Breast

    Directory of Open Access Journals (Sweden)

    Brittany Busse

    2014-01-01

    Full Text Available Prosthetic implants are frequently used for breast augmentation and breast reconstruction following mastectomy. Unfortunately, long-term aesthetic results of prosthetic breast restoration may be hindered by complications such as rippling, capsular contracture, and implant malposition. The advent of use of acellular dermal matrices has greatly improved the outcomes of prosthetic breast reconstruction. We describe a case of rippling deformity of breast that was treated using an acellular dermal matrix product, AlloMax. The patient presented with visible rippling of bilateral prosthetic breast implants as well as significant asymmetry of the breasts after multiple excisional biopsies for right breast ductal carcinoma in situ. A 6×10 cm piece of AlloMax was placed on the medial aspect of each breast between the implant and the skin flap. Follow-up was performed at 1 week, 3 months, and 1 year following the procedure. The patient recovered well from the surgery and there were no complications. At her first postoperative follow-up the patient was extremely satisfied with the result. At her 3-month and 1-year follow-up she had no recurrence of her previous deformity and no new deformity.

  20. Composition of acellular pertussis and combination vaccines: a general review.

    Science.gov (United States)

    Jadhav, S S; Gairola, S

    1999-06-01

    Since the development and introduction of the acellular pertussis vaccine in Japan in the early eighties, we have come a long way in using this component in combination with other vaccines. However, the basic problem in development of an effective and safe pertussis vaccine is that the antigens to induce complete protection against clinical pertussis and the precise mechanism by which pertussis vaccine confers immunity is yet unknown. Hence, the composition of future acellular pertussis vaccine remains an open issue. Recently, acellular pertussis vaccine has been licensed for the booster doses in the U.S.A. and for primary immunization of infants in Italy and Germany. A multicentric trial has been carried out to compare the serological response and adverse reactions of 13 acellular pertussis vaccines. These vaccines contained one or more of the four components, i.e. FHA, PT, 69 kDa OMP and fimbriae. All vaccines were associated with substantially fewer and less adverse reactions and were more immunogenic with respect to antibodies against the added antigens. DTP vaccines in the near future will have combinations of other components and the key antigen for combination will be acellular pertussis component which is going to replace whole cell pertussis component in DTP vaccines. In view of this, manufacturers like ourselves from the developing countries are still groping in the dark, uncertain whether we should have a single component acellular pertussis vaccine or multicomponent one. This will have a major impact on the cost of production, the final cost of the combination vaccines and the regulatory issues that we will have to tackle in view of the recent thinking on harmonization in the pharmaceutical industry. Copyright 1999 The International Association for Biologicals.

  1. Detection of HOCl-mediated protein oxidation products in the extracellular matrix of human atherosclerotic plaques

    DEFF Research Database (Denmark)

    Woods, Alan A; Linton, Stuart M; Davies, Michael Jonathan

    2003-01-01

    for 83-96% of the total oxidized protein side-chain products detected in these plaques. Oxidation of matrix components extracted from healthy artery tissue, and model proteins, with reagent HOCl is shown to give rise to a similar pattern of products to those detected in advanced human lesions......Oxidation is believed to play a role in atherosclerosis. Oxidized lipids, sterols and proteins have been detected in early, intermediate and advanced human lesions at elevated levels. The spectrum of oxidized side-chain products detected on proteins from homogenates of advanced human lesions has...... by activated monocytes (and possibly macrophages) and is a highly basic protein, it would be expected to associate with polyanions such as the glycosaminoglycans of the extracellular matrix, and might result in damage being localized at such sites. In this study proteins extracted from extracellular matrix...

  2. [Effect of two different acellular lung matrices on α-SMA expression in A549 cells].

    Science.gov (United States)

    Chen, C; Wang, Z Y; Weng, J; Wang, Z B; Mei, J; Du, X H; Wang, L

    2017-01-24

    Objective: To explore the effect of acellular normal and fibrotic lung matrices on alpha smooth muscle actin (α-SMA) expression in human lung adenocarcinoma cell line A549. Methods: Twenty adult SD rats were randomly divided into normal group and idiopathic pulmonary fibrosis(IPF)group (n=10 each). The pulmonary fibrosis was induced by Bleomycin. Normal and fibrotic decellularized lungs were made, then sections with 500 μm thick were cut by a standard Vibratome. None scaffold was set as control group. A549 cells were seeded dropwise into different slices (normal and fibrotic scaffolds), and cultured for one week in vitro. The expression of α-SMA was measured by immunofluorescence staining and quantitative real time polymerase chain reaction (qRT-PCR). Results: In control group, the expression of α-SMA protein was positive in A549 cells by immunofluorescence staining. However, it expressed weakly both in normal and fibrotic scaffold group, and the fluorescence intensity in fibrotic scaffold group was significant lower than that in normal group (PSMA mRNA in normal and fibrotic scaffold group were (0.70±0.11) and (0.55±0.12), which were significant lower than that of control group (1.28±0.21) (PSMA mRNA in fibrotic scaffold group was decreased compared to that in normal scaffold group (PSMA in human lung adenocarcinoma cell line A549. It may inhibit the movement of A549 cells in acellular normal and fibrotic lung matrices, especially in acellular fibrotic lung scaffold.

  3. Microrheology and ROCK signaling of human endothelial cells embedded in a 3D matrix.

    Science.gov (United States)

    Panorchan, Porntula; Lee, Jerry S H; Kole, Thomas P; Tseng, Yiider; Wirtz, Denis

    2006-11-01

    Cell function is profoundly affected by the geometry of the extracellular environment confining the cell. Whether and how cells plated on a two-dimensional matrix or embedded in a three-dimensional (3D) matrix mechanically sense the dimensionality of their environment is mostly unknown, partly because individual cells in an extended matrix are inaccessible to conventional cell-mechanics probes. Here we develop a functional assay based on multiple particle tracking microrheology coupled with ballistic injection of nanoparticles to measure the local intracellular micromechanical properties of individual cells embedded inside a matrix. With our novel assay, we probe the mechanical properties of the cytoplasm of individual human umbilical vein endothelial cells (HUVECs) embedded in a 3D peptide hydrogel in the presence or absence of vascular endothelial growth factor (VEGF). We found that VEGF treatment, which enhances endothelial migration, increases the compliance and reduces the elasticity of the cytoplasm of HUVECs in a matrix. This VEGF-induced softening response of the cytoplasm is abrogated by specific Rho-kinase (ROCK) inhibition. These results establish combined particle-tracking microrheology and ballistic injection as the first method able to probe the micromechanical properties and mechanical response to agonists and/or drug treatments of individual cells inside a matrix. These results suggest that ROCK plays an essential role in the regulation of the intracellular mechanical response to VEGF of endothelial cells in a 3D matrix.

  4. Follow-up review on the long-term effect of composite transplantation of allogeneic acellular dermal matrix and split thickness skin autograft%异体脱细胞真皮基质加自体刃厚皮复合移植远期随访评价

    Institute of Scientific and Technical Information of China (English)

    潘云川; 徐家钦; 袁素; 梁尊鸿; 陈思环; 陈茹妹; 林思燕

    2010-01-01

    目的 评价异体ADM+自体刃厚皮复合移植的临床远期效果.方法 选择2001年3月-2008年10月,笔者单位收治的19例行异体ADM+自体刃厚皮复合移植患者为复合移植组(34个创面),同期9例行自体刃厚皮移植患者为对照组(11个创面).患者术后均随访2年以上.随访时,在曼彻斯特瘢痕量表的基础上设计随访对象评估表,评估移植皮肤的颜色、平整度、质地、挛缩、感觉、并发症情况,分值1~4分,得分越高、情况越差;采用温哥华瘢痕量表评估供皮区瘢痕形成情况;发放问卷调查患者满意度、移植期内健康记录;组织病理学方法观察其中4例患者皮肤组织结构.采用中立位法描述术前、术后及随访时患者关节活动范围.对数据进行非参数秩和检验、t检验或x2检验.结果 (1)复合移植组皮肤平整度、挛缩、质地评分分别为(1.6±0.5)、(1.8±0.8)、(1.5±0.8)分,显著低于对照组的(2.0±0.7)、(2.2±0.9)、(2.3±0.7)分(Z值分别为-2.058、-2.220、-2.323,P值均小于0.05);2组皮肤颜色、感觉、并发症评分结果相近(Z值分别为-0.628、-0.428、-2.520,P值均大于0.05).(2)复合移植组仅1个供皮区部分区域有轻度瘢痕.(3)复合移植组和对照组患者在疼痛、瘙痒和满意度方面比较,差异均无统计学意义(x2值分别为0.187、0.019、2.628,P值均大于0.05).(4)病理结果显示,手部复合移植后2年可见神经纤维结构,ADM在受体内未引起强烈的炎症反应.(5)复合移植组11处关节部位功能得到恢复或改善,另2处需再次手术.对照组2处关节部位均明显挛缩.结论 异体ADM+自体刃厚皮复合移植在防止瘢痕挛缩,改善功能及外观方面效果明显,长期存留于成人和儿童患者体内均未出现安全问题.%Objective To review the long-term clinical effect of composite transplantation of allogeneic acellular dermal matrix (ADM) and split thickness skin autograft (STSG). Methods

  5. 脱细胞组织工程真皮基质修复供皮区创面的临床观察%Clinical observation on repairing of wounds of skin graft donor site with acellular tissue engineering dermal matrix

    Institute of Scientific and Technical Information of China (English)

    邱学文; 王甲汉; 王颖; 刘亮; 吴起; 马军

    2013-01-01

    Objective To evaluate the clinical efficacy of acellular tissue engineering dermal matrix (ATDM) in repairing wounds of skin graft donor site.Methods Sixty patients with burn or chronic wounds hospitalized from January 2011 to April 2012 received autologous skin grafting.One wound [with size larger than 55 cm2,and thickness of (0.33 ± 0.03) mm] out of multiple skin graft donor sites of every patient was selected,and it was divided into two parts in accordance with self-control principle.A part of wound close to the wound edge with diameter of 5 cm was taken as trial area (treated with ATDM),and the remaining wound was taken as control area (treated with vaseline gauze) according to the random number table.Blood and urine routine,liver and kidney function,and levels of IgG and IgM in blood of patients were measured one day before operation and on the 1 st day after wound healing.Vital signs of patients were recorded on the operation day and the wound healing day.Gross condition of the wounds was observed during dressing change.Wound healing time was recorded.The healed wound was observed histologically.Data were processed with Logrank test or t test.Results Leucocyte count was lowered on the 1st day after wound healing [(7.1 ± 1.2) × 109/L] as compared with that one day before operation [(10.1 ± 1.5) ×109/L,t =-12.10,P <0.01].The differences were not statistically significant in red blood cell count,haemoglobin level,platelet count,urine routine,levels of indexes of liver and kidney function,levels of IgG and IgM in blood between one day before operation and the 1st day after wound healing,or in vital signs (including body temperature,pulse,respiration,systolic pressure,and diastolic pressure) between the operation day and the wound healing day (with t values from-1.43 to 1.88,P values all above 0.05).No adverse effects such as abnormal exudation,itching,redness and swelling,and exanthema were observed in the wound.The median wound healing time in trial area

  6. Degenerated human intervertebral discs contain autoantibodies against extracellular matrix proteins

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    S Capossela

    2014-04-01

    Full Text Available Degeneration of intervertebral discs (IVDs is associated with back pain and elevated levels of inflammatory cells. It has been hypothesised that discogenic pain is a direct result of vascular and neural ingrowth along annulus fissures, which may expose the avascular nucleus pulposus (NP to the systemic circulation and induce an autoimmune reaction. In this study, we confirmed our previous observation of antibodies in human degenerated and post-traumatic IVDs cultured in vitro. We hypothesised that the presence of antibodies was due to an autoimmune reaction against specific proteins of the disc. Furthermore we identified antigens which possibly trigger an autoimmune response in degenerative disc diseases. We demonstrated that degenerated and post-traumatic IVDs contain IgG antibodies against typical extracellular proteins of the disc, particularly proteins of the NP. We identified IgGs against collagen type II and aggrecan, confirming an autoimmune reaction against the normally immune privileged NP. We also found specific IgGs against collagens types I and V, but not against collagen type III. In conclusion, this study confirmed the association between disc degeneration and autoimmunity, and may open the avenue for future studies on developing prognostic, diagnostic and therapy-monitoring markers for degenerative disc diseases.

  7. Degenerated human intervertebral discs contain autoantibodies against extracellular matrix proteins.

    Science.gov (United States)

    Capossela, S; Schläfli, P; Bertolo, A; Janner, T; Stadler, B M; Pötzel, T; Baur, M; Stoyanov, J V

    2014-04-04

    Degeneration of intervertebral discs (IVDs) is associated with back pain and elevated levels of inflammatory cells. It has been hypothesised that discogenic pain is a direct result of vascular and neural ingrowth along annulus fissures, which may expose the avascular nucleus pulposus (NP) to the systemic circulation and induce an autoimmune reaction. In this study, we confirmed our previous observation of antibodies in human degenerated and post-traumatic IVDs cultured in vitro. We hypothesised that the presence of antibodies was due to an autoimmune reaction against specific proteins of the disc. Furthermore we identified antigens which possibly trigger an autoimmune response in degenerative disc diseases. We demonstrated that degenerated and post-traumatic IVDs contain IgG antibodies against typical extracellular proteins of the disc, particularly proteins of the NP. We identified IgGs against collagen type II and aggrecan, confirming an autoimmune reaction against the normally immune privileged NP. We also found specific IgGs against collagens types I and V, but not against collagen type III. In conclusion, this study confirmed the association between disc degeneration and autoimmunity, and may open the avenue for future studies on developing prognostic, diagnostic and therapy-monitoring markers for degenerative disc diseases.

  8. Visualization of extracellular matrix components within sectioned Salmonella biofilms on the surface of human gallstones.

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    Joanna M Marshall

    Full Text Available Chronic carriage of Salmonella Typhi is mediated primarily through the formation of bacterial biofilms on the surface of cholesterol gallstones. Biofilms, by definition, involve the formation of a bacterial community encased within a protective macromolecular matrix. Previous work has demonstrated the composition of the biofilm matrix to be complex and highly variable in response to altered environmental conditions. Although known to play an important role in bacterial persistence in a variety of contexts, the Salmonella biofilm matrix remains largely uncharacterized under physiological conditions. Initial attempts to study matrix components and architecture of the biofilm matrix on gallstone surfaces were hindered by the auto-fluorescence of cholesterol. In this work we describe a method for sectioning and direct visualization of extracellular matrix components of the Salmonella biofilm on the surface of human cholesterol gallstones and provide a description of the major matrix components observed therein. Confocal micrographs revealed robust biofilm formation, characterized by abundant but highly heterogeneous expression of polysaccharides such as LPS, Vi and O-antigen capsule. CsgA was not observed in the biofilm matrix and flagellar expression was tightly restricted to the biofilm-cholesterol interface. Images also revealed the presence of preexisting Enterobacteriaceae encased within the structure of the gallstone. These results demonstrate the use and feasibility of this method while highlighting the importance of studying the native architecture of the gallstone biofilm. A better understanding of the contribution of individual matrix components to the overall biofilm structure will facilitate the development of more effective and specific methods to disrupt these bacterial communities.

  9. Visualization of Extracellular Matrix Components within Sectioned Salmonella Biofilms on the Surface of Human Gallstones

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    Marshall, Joanna M.; Flechtner, Alan D.; La Perle, Krista M.; Gunn, John S.

    2014-01-01

    Chronic carriage of Salmonella Typhi is mediated primarily through the formation of bacterial biofilms on the surface of cholesterol gallstones. Biofilms, by definition, involve the formation of a bacterial community encased within a protective macromolecular matrix. Previous work has demonstrated the composition of the biofilm matrix to be complex and highly variable in response to altered environmental conditions. Although known to play an important role in bacterial persistence in a variety of contexts, the Salmonella biofilm matrix remains largely uncharacterized under physiological conditions. Initial attempts to study matrix components and architecture of the biofilm matrix on gallstone surfaces were hindered by the auto-fluorescence of cholesterol. In this work we describe a method for sectioning and direct visualization of extracellular matrix components of the Salmonella biofilm on the surface of human cholesterol gallstones and provide a description of the major matrix components observed therein. Confocal micrographs revealed robust biofilm formation, characterized by abundant but highly heterogeneous expression of polysaccharides such as LPS, Vi and O-antigen capsule. CsgA was not observed in the biofilm matrix and flagellar expression was tightly restricted to the biofilm-cholesterol interface. Images also revealed the presence of preexisting Enterobacteriaceae encased within the structure of the gallstone. These results demonstrate the use and feasibility of this method while highlighting the importance of studying the native architecture of the gallstone biofilm. A better understanding of the contribution of individual matrix components to the overall biofilm structure will facilitate the development of more effective and specific methods to disrupt these bacterial communities. PMID:24551241

  10. Lack of association between mannose binding lectin and antibody responses after acellular pertussis vaccinations.

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    Kirsi Gröndahl-Yli-Hannuksela

    Full Text Available BACKGROUND: Mannose-binding lectin (MBL is one of the key molecules in innate immunity and its role in human vaccine responses is poorly known. This study aimed to investigate the possible association of MBL polymorphisms with antibody production after primary and booster vaccinations with acellular pertussis vaccines in infants and adolescents. METHODOLOGY/PRINCIPAL FINDINGS: Five hundred and sixty eight subjects were included in this study. In the adolescent cohort 355 subjects received a dose of diphtheria and tetanus toxoids and acellular pertussis (dTpa vaccine ten years previously. Follow-up was performed at 3, 5 and 10 years. Infant cohort consisted of 213 subjects, who had received three primary doses of DTaP vaccine at 3, 5, and 12 months of age according to Finnish immunization program. Blood samples were collected before the vaccinations at 2,5 months of age and after the vaccinations at 13 months and 2 years of age. Concentrations of IgG antibodies to pertussis toxin, filamentous hemagglutinin, and pertactin and antibodies to diphtheria and tetanus toxoids were measured by standardized enzyme-linked immunosorbant assay. Single nucleotide polymorphisms of MBL2 gene exon1 (codons 52, 54, 57 were examined. MBL serum concentration was also measured from the adolescent cohort. No association was found with MBL2 exon 1 polymorphisms and antibody responses against vaccine antigens, after primary and booster dTpa vaccination. CONCLUSIONS: This study indicates that MBL polymorphisms do not affect the production and persistence of antibodies after acellular pertussis vaccination. Our finding also suggests that MBL might not be involved in modulating antibody responses to the vaccines made of purified bacterial proteins.

  11. Development of biomimetic nanocomposites as bone extracellular matrix for human osteoblastic cells.

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    Bhowmick, Arundhati; Mitra, Tapas; Gnanamani, Arumugam; Das, Manas; Kundu, Patit Paban

    2016-05-05

    Here, we have developed biomimetic nanocomposites containing chitosan, poly(vinyl alcohol) and nano-hydroxyapatite-zinc oxide as bone extracellular matrix for human osteoblastic cells and characterized by Fourier transform infrared spectroscopy, powder X-ray diffraction. Scanning electron microscopy images revealed interconnected macroporous structures. Moreover, in this study, the problem related to fabricating a porous composite with good mechanical strength has been resolved by incorporating 5wt% of nano-hydroxyapatite-zinc oxide into chitosan-poly(vinyl alcohol) matrix; the present composite showed high tensile strength (20.25MPa) while maintaining appreciable porosity (65.25%). These values are similar to human cancellous bone. These nanocomposites also showed superior water uptake, antimicrobial and biodegradable properties than the previously reported results. Compatibility with human blood and pH was observed, indicating nontoxicity of these materials to the human body. Moreover, proliferation of osteoblastic MG-63 cells onto the nanocomposites was also observed without having any negative effect.

  12. Acellular Nerve Allografts in Peripheral Nerve Regeneration: A Comparative Study

    Science.gov (United States)

    Moore, Amy M.; MacEwan, Matthew; Santosa, Katherine B.; Chenard, Kristofer E.; Ray, Wilson Z.; Hunter, Daniel A.; Mackinnon, Susan E.; Johnson, Philip J.

    2011-01-01

    Background Processed nerve allografts offer a promising alternative to nerve autografts in the surgical management of peripheral nerve injuries where short deficits exist. Methods Three established models of acellular nerve allograft (cold-preserved, detergent-processed, and AxoGen® -processed nerve allografts) were compared to nerve isografts and silicone nerve guidance conduits in a 14 mm rat sciatic nerve defect. Results All acellular nerve grafts were superior to silicone nerve conduits in support of nerve regeneration. Detergent-processed allografts were similar to isografts at 6 weeks post-operatively, while AxoGen®-processed and cold-preserved allografts supported significantly fewer regenerating nerve fibers. Measurement of muscle force confirmed that detergent-processed allografts promoted isograft-equivalent levels of motor recovery 16 weeks post-operatively. All acellular allografts promoted greater amounts of motor recovery compared to silicone conduits. Conclusions These findings provide evidence that differential processing for removal of cellular constituents in preparing acellular nerve allografts affects recovery in vivo. PMID:21660979

  13. Postsynaptic density protein 95 in the striosome and matrix compartments of the human neostriatum.

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    Ryoma eMorigaki

    2015-11-01

    Full Text Available The human neostriatum consists of two functional subdivisions referred to as the striosome (patch and matrix compartments. The striosome-matrix dopamine systems play a central role in cortico-thalamo-basal ganglia circuits, and their involvement is thought to underlie the genesis of multiple movement and behavioral disorders, and of drug addiction. Human neuropathology also has shown that striosomes and matrix have differential vulnerability patterns in several striatal neurodegenerative diseases. Postsynaptic density protein 95 (PSD-95, also known as DLG4, is a major scaffolding protein in the postsynaptic densities of dendritic spines. PSD-95 is now known to negatively regulate not only N-methyl-D-aspartate glutamate signaling, but also dopamine D1 signals at sites of postsynaptic transmission. Accordingly, a neuroprotective role for PSD-95 against dopamine D1 receptor (D1R-mediated neurotoxicity in striatal neurodegeneration also has been suggested. Here, we used a highly sensitive immunohistochemistry technique to show that in the human neostriatum, PSD-95 is differentially concentrated in the striosome and matrix compartments, with a higher density of PSD-95 labeling in the matrix compartment than in the striosomes. This compartment-specific distribution of PSD-95 was strikingly complementary to that of D1R. In addition to the possible involvement of PSD-95-mediated synaptic function in compartment-specific dopamine signals, we suggest that the striosomes might be more susceptible to D1R-mediated neurotoxicity than the matrix compartment. This notion may provide new insight into the compartment-specific vulnerability of MSNs in striatal neurodegeneration.

  14. Human epidermal keratinocyte cell response on integrin-specific artificial extracellular matrix proteins.

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    Tjin, Monica Suryana; Chua, Alvin Wen Choong; Ma, Dong Rui; Lee, Seng Teik; Fong, Eileen

    2014-08-01

    Cell-matrix interactions play critical roles in regulating cellular behavior in wound repair and regeneration of the human skin. In particular, human skin keratinocytes express several key integrins such as alpha5beta1, alpha3beta1, and alpha2beta1 for binding to the extracellular matrix (ECM) present in the basement membrane in uninjured skin. To mimic these key integrin-ECM interactions, artificial ECM (aECM) proteins containing functional domains derived from laminin 5, type IV collagen, fibronectin, and elastin are prepared. Human skin keratinocyte cell responses on the aECM proteins are specific to the cell-binding domain present in each construct. Keratinocyte attachment to the aECM protein substrates is also mediated by specific integrin-material interactions. In addition, the aECM proteins are able to support the proliferation of keratinocyte stem cells, demonstrating their promise for use in skin tissue engineering.

  15. Matrix Stiffness Corresponding to Strictured Bowel Induces a Fibrogenic Response in Human Colonic Fibroblasts

    Science.gov (United States)

    Johnson, Laura A.; Rodansky, Eva S.; Sauder, Kay L.; Horowitz, Jeffrey C.; Mih, Justin D.; Tschumperlin, Daniel J.; Higgins, Peter D.

    2013-01-01

    Background Crohn’s disease is characterized by repeated cycles of inflammation and mucosal healing which ultimately progress to intestinal fibrosis. This inexorable progression towards fibrosis suggests that fibrosis becomes inflammation-independent and auto-propagative. We hypothesized that matrix stiffness regulates this auto-propagation of intestinal fibrosis. Methods The stiffness of fresh ex vivo samples from normal human small intestine, Crohn’s disease strictures, and the unaffected margin were measured with a microelastometer. Normal human colonic fibroblasts were cultured on physiologically normal or pathologically stiff matrices corresponding to the physiological stiffness of normal or fibrotic bowel. Cellular response was assayed for changes in cell morphology, α-smooth muscle actin (αSMA) staining, and gene expression. Results Microelastometer measurements revealed a significant increase in colonic tissue stiffness between normal human colon and Crohn’s strictures as well as between the stricture and adjacent tissue margin. In Ccd-18co cells grown on stiff matrices corresponding to Crohn’s strictures, cellular proliferation increased. Pathologic stiffness induced a marked change in cell morphology and increased αSMA protein expression. Growth on a stiff matrix induced fibrogenic gene expression, decreased matrix metalloproteinase and pro-inflammatory gene expression, and was associated with nuclear localization of the transcriptional cofactor MRTF-A. Conclusions Matrix stiffness, representative of the pathological stiffness of Crohn’s strictures, activates human colonic fibroblasts to a fibrogenic phenotype. Matrix stiffness affects multiple pathways suggesting the mechanical properties of the cellular environment are critical to fibroblast function and may contribute to autopropagation of intestinal fibrosis in the absence of inflammation, thereby contributing to the intractable intestinal fibrosis characteristic of Crohn’s disease. PMID

  16. Research and Application of Human Capital Strategic Classification Tool: Human Capital Classification Matrix Based on Biological Natural Attribute

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    Yong Liu

    2014-12-01

    Full Text Available In order to study the causes of weak human capital structure strategic classification management in China, we analyze that enterprises around the world face increasingly difficult for human capital management. In order to provide strategically sound answers, the HR managers need the critical information provided by the right technology processing and analytical tools. In this study, there are different types and levels of human capital in formal organization management, which is not the same contribution to a formal organization. An important guarantee for sustained and healthy development of the formal or informal organization is lower human capital risk. To resist this risk is primarily dependent on human capital hedge force and appreciation force in value, which is largely dependent on the strategic value of the performance of senior managers. Based on the analysis of high-level managers perspective, we also discuss the value and configuration of principles and methods to be followed in human capital strategic classification based on Boston Consulting Group (BCG matrix and build Human Capital Classification (HCC matrix based on biological natural attribute to effectively realize human capital structure strategic classification.

  17. Interphotoreceptor matrix-poly(ϵ-caprolactone composite scaffolds for human photoreceptor differentiation

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    Petr Baranov

    2014-10-01

    Full Text Available Tissue engineering has been widely applied in different areas of regenerative medicine, including retinal regeneration. Typically, artificial biopolymers require additional surface modification (e.g. with arginine–glycine–aspartate-containing peptides or adsorption of protein, such as fibronectin, before cell seeding. Here, we describe an alternative approach for scaffold design: the manufacture of hybrid interphotoreceptor matrix-poly (ϵ-caprolactone scaffolds, in which the insoluble extracellular matrix of the retina is incorporated into a biodegradable polymer well suited for transplantation. The incorporation of interphotoreceptor matrix did not change the topography of polycaprolactone film, although it led to a slight increase in hydrophilic properties (water contact angle measurements. This hybrid scaffold provided sufficient stimuli for human retinal progenitor cell adhesion and inhibited proliferation, leading to differentiation toward photoreceptor cells (expression of Crx, Nrl, rhodopsin, ROM1. This scaffold may be used for transplantation of retinal progenitor cells and their progeny to treat retinal degenerative disorders.

  18. In vitro human periodontal ligament-like tissue formation with porous poly-L-lactide matrix

    Energy Technology Data Exchange (ETDEWEB)

    Liao, Wen [Graduate School of Dentistry, Department of Orthodontics, Osaka Dental University, 8-1 Kuzuha-hanazono-cho, Hirakata-shi, Osaka-fu 573-1121 (Japan); Okada, Masahiro [Department of Biomaterials, Osaka Dental University, 8-1 Kuzuha-hanazono-cho, Hirakata-shi, Osaka-fu 573-1121 (Japan); Sakamoto, Fumito; Okita, Naoya [Graduate School of Dentistry, Department of Orthodontics, Osaka Dental University, 8-1 Kuzuha-hanazono-cho, Hirakata-shi, Osaka-fu 573-1121 (Japan); Inami, Kaoru; Nishiura, Aki [Department of Orthodontics, Osaka Dental University, 8-1 Kuzuha-hanazono-cho, Hirakata-shi, Osaka-fu 573-1121 (Japan); Hashimoto, Yoshiya, E-mail: yoshiya@cc.osaka-dent.ac.jp [Department of Biomaterials, Osaka Dental University, 8-1 Kuzuha-hanazono-cho, Hirakata-shi, Osaka-fu 573-1121 (Japan); Matsumoto, Naoyuki [Department of Orthodontics, Osaka Dental University, 8-1 Kuzuha-hanazono-cho, Hirakata-shi, Osaka-fu 573-1121 (Japan)

    2013-08-01

    This study aimed to establish an in vitro human periodontal ligament-like tissue (HPdLLT) by three-dimensional culturing of human periodontal ligament fibroblasts (HPdLFs) in a porous poly-L-lactide (PLLA) matrix modified hydrophilically with ammonia solution. After ammonia modification, the surface roughness and culture-medium-soaking-up ability of the PLLA matrix increased, whereas the contact angle of water drops decreased. The thickness, porosity, and pore size of the PLLA matrix were 400 ± 50 μm, 83.3%, and 75–150 μm, respectively. HPdLFs (1 × 10{sup 5} cells) were seeded on the modified PLLA matrix and centrifuged to facilitate seeding into its interior and cultured for 14 days. Scanning electron microscope (SEM) observation, proliferation assay, picrosirius-red staining, and real-time polymerase chain reaction (RT-PCR) for type-1 collagen (COL1), periodontal ligament associated protein-1 (PLAP-1), fibroblast growth factor-2 (FGF-2), and alkaline phosphatase (ALP) mRNA were conducted on days 1, 3, 7, and 14. HPdLFs were observed entirely from the surface to the rear side of the matrix. Cell proliferation analysis, SEM observation, and picrosirius-red staining showed both progressive growth of 3D-cultured HPdLFs and extracellular matrix maturation by the secretion of COL1 and type 3 collagen (COL3) from days 1 to 14. Expressions of COL1, PLAP-1, and FGF-2 mRNA suggested the formation of cellular components and supplementation of extracellular components. Expressions of ALP, COL1, and PLAP-1 mRNA suggested the osteogenic potential of the HPdLLT. The results indicated in vitro HPdLLT formation, and it could be used in future periodontal ligament tissue engineering to achieve optimal periodontal regeneration. - Highlights: • First report on ammonia treated PLLA matrix for in vitro human periodontal ligament-like tissue generation. • Good combination of matrix thickness, pore size, and porosity. • Biodegradable PLLA is also possible to be used in vivo.

  19. In vitro human periodontal ligament-like tissue formation with porous poly-L-lactide matrix.

    Science.gov (United States)

    Liao, Wen; Okada, Masahiro; Sakamoto, Fumito; Okita, Naoya; Inami, Kaoru; Nishiura, Aki; Hashimoto, Yoshiya; Matsumoto, Naoyuki

    2013-08-01

    This study aimed to establish an in vitro human periodontal ligament-like tissue (HPdLLT) by three-dimensional culturing of human periodontal ligament fibroblasts (HPdLFs) in a porous poly-L-lactide (PLLA) matrix modified hydrophilically with ammonia solution. After ammonia modification, the surface roughness and culture-medium-soaking-up ability of the PLLA matrix increased, whereas the contact angle of water drops decreased. The thickness, porosity, and pore size of the PLLA matrix were 400±50 μm, 83.3%, and 75-150 μm, respectively. HPdLFs (1×10(5) cells) were seeded on the modified PLLA matrix and centrifuged to facilitate seeding into its interior and cultured for 14 days. Scanning electron microscope (SEM) observation, proliferation assay, picrosirius-red staining, and real-time polymerase chain reaction (RT-PCR) for type-1 collagen (COL1), periodontal ligament associated protein-1 (PLAP-1), fibroblast growth factor-2 (FGF-2), and alkaline phosphatase (ALP) mRNA were conducted on days 1, 3, 7, and 14. HPdLFs were observed entirely from the surface to the rear side of the matrix. Cell proliferation analysis, SEM observation, and picrosirius-red staining showed both progressive growth of 3D-cultured HPdLFs and extracellular matrix maturation by the secretion of COL1 and type 3 collagen (COL3) from days 1 to 14. Expressions of COL1, PLAP-1, and FGF-2 mRNA suggested the formation of cellular components and supplementation of extracellular components. Expressions of ALP, COL1, and PLAP-1 mRNA suggested the osteogenic potential of the HPdLLT. The results indicated in vitro HPdLLT formation, and it could be used in future periodontal ligament tissue engineering to achieve optimal periodontal regeneration.

  20. Monocytes increase human cardiac myofibroblast-mediated extracellular matrix remodeling through TGF-β1.

    Science.gov (United States)

    Mewhort, Holly E M; Lipon, Brodie D; Svystonyuk, Daniyil A; Teng, Guoqi; Guzzardi, David G; Silva, Claudia; Yong, V Wee; Fedak, Paul W M

    2016-03-15

    Following myocardial infarction (MI), cardiac myofibroblasts remodel the extracellular matrix (ECM), preventing mechanical complications. However, prolonged myofibroblast activity leads to dysregulation of the ECM, maladaptive remodeling, fibrosis, and heart failure (HF). Chronic inflammation is believed to drive persistent myofibroblast activity; however, the mechanisms are unclear. We assessed the influence of peripheral blood monocytes on human cardiac myofibroblast activity in a three-dimensional (3D) ECM microenvironment. Human cardiac myofibroblasts isolated from surgical biopsies of the right atrium and left ventricle were seeded into 3D collagen matrices. Peripheral blood monocytes were isolated from healthy human donors and cocultured with myofibroblasts. Monocytes increased myofibroblast activity measured by collagen gel contraction (baseline: 57.6 ± 5.9% vs. coculture: 65.2 ± 7.1% contraction; P matrix metalloproteinase 9 compared with baseline (122.9 ± 10.1 pg/ml and 3,496.0 ± 190.4 pg/ml, respectively, vs. 21.5 ± 16.3 pg/ml and 183.3 ± 43.9 pg/ml; P matrix. Peripheral blood monocyte interaction with human cardiac myofibroblasts stimulates myofibroblast activity through release of TGF-β1. These data implicate inflammation as a potential driver of cardiac fibrosis.

  1. Modulating notochordal differentiation of human induced pluripotent stem cells using natural nucleus pulposus tissue matrix.

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    Yongxing Liu

    Full Text Available Human induced pluripotent stem cells (hiPSCs can differentiate into notochordal cell (NC-like cells when cultured in the presence of natural porcine nucleus pulposus (NP tissue matrix. The method promises massive production of high-quality, functional cells to treat degenerative intervertebral discs (IVDs. Based on our previous work, we further examined the effect of cell-NP matrix contact and culture medium on the differentiation, and further assessed the functional differentiation ability of the generated NC-like. The study showed that direct contact between hiPSCs and NP matrix can promote the differentiation yield, whilst both the contact and non-contact cultures can generate functional NC-like cells. The generated NC-like cells are highly homogenous regarding the expression of notochordal marker genes. A culture medium containing a cocktail of growth factors (FGF, EGF, VEGF and IGF-1 also supported the notochordal differentiation in the presence of NP matrix. The NC-like cells showed excellent functional differentiation ability to generate NP-like tissue which was rich in aggrecan and collagen type II; and particularly, the proteoglycan to collagen content ratio was as high as 12.5-17.5 which represents a phenotype close to NP rather than hyaline cartilage. Collectively, the present study confirmed the effectiveness and flexibility of using natural NP tissue matrix to direct notochordal differentiation of hiPSCs, and the potential of using the generated NC-like cells for treating IVD degeneration.

  2. Neutrophil-Derived MMP-8 Drives AMPK-Dependent Matrix Destruction in Human Pulmonary Tuberculosis.

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    Catherine W M Ong

    2015-05-01

    Full Text Available Pulmonary cavities, the hallmark of tuberculosis (TB, are characterized by high mycobacterial load and perpetuate the spread of M. tuberculosis. The mechanism of matrix destruction resulting in cavitation is not well defined. Neutrophils are emerging as key mediators of TB immunopathology and their influx are associated with poor outcomes. We investigated neutrophil-dependent mechanisms involved in TB-associated matrix destruction using a cellular model, a cohort of 108 patients, and in separate patient lung biopsies. Neutrophil-derived NF-kB-dependent matrix metalloproteinase-8 (MMP-8 secretion was up-regulated in TB and caused matrix destruction both in vitro and in respiratory samples of TB patients. Collagen destruction induced by TB infection was abolished by doxycycline, a licensed MMP inhibitor. Neutrophil extracellular traps (NETs contain MMP-8 and are increased in samples from TB patients. Neutrophils lined the circumference of human pulmonary TB cavities and sputum MMP-8 concentrations reflected TB radiological and clinical disease severity. AMPK, a central regulator of catabolism, drove neutrophil MMP-8 secretion and neutrophils from AMPK-deficient patients secrete lower MMP-8 concentrations. AMPK-expressing neutrophils are present in human TB lung biopsies with phospho-AMPK detected in nuclei. These data demonstrate that neutrophil-derived MMP-8 has a key role in the immunopathology of TB and is a potential target for host-directed therapy in this infectious disease.

  3. 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.

  4. Human 2'-phosphodiesterase localizes to the mitochondrial matrix with a putative function in mitochondrial RNA turnover

    DEFF Research Database (Denmark)

    Poulsen, Jesper Buchhave; Andersen, Kasper Røjkjær; Kjær, Karina Hansen

    2011-01-01

    . Interestingly, 2′-PDE shares both functionally and structurally characteristics with the CCR4-type exonuclease–endonuclease–phosphatase family of deadenylases. Here we show that 2′-PDE locates to the mitochondrial matrix of human cells, and comprise an active 3′–5′ exoribonuclease exhibiting a preference...... a role in the cellular immune system, may also function in mitochondrial RNA turnover....

  5. Matrix-specified differentiation of human decidua parietalis placental stem cells

    OpenAIRE

    Sridharan, Indumathi; Kim, Taeyoung; Strakova, Zuzana; Wang, Rong

    2013-01-01

    To create suitable biological scaffolds for tissue engineering and cell therapeutics, it is essential to understand the matrix-mediated specification of stem cell differentiation. To this end, we studied the effect of collagen type I on stem cell lineage specification. We altered the properties of collagen type I by incorporating carbon nanotubes (CNT). The collagen-CNT composite material was stiffer with thicker fibers and longer D-period. Human decidua parietalis stem cells (hdpPSC) were fo...

  6. Tetanus–diphtheria–acellular pertussis vaccination for adults: an update

    Science.gov (United States)

    2017-01-01

    Although tetanus and diphtheria have become rare in developed countries, pertussis is still endemic in some developed countries. These are vaccine-preventable diseases and vaccination for adults is important to prevent the outbreak of disease. Strategies for tetanus, diphtheria, and pertussis vaccines vary from country to country. Each country needs to monitor consistently epidemiology of the diseases and changes vaccination policies accordingly. Recent studies showed that tetanus–diphtheria–acellular pertussis vaccine for adults is effective and safe to prevent pertussis disease in infants. However, vaccine coverage still remains low than expected and seroprevalence of protective antibodies levels for tetanus, diphtheria, and pertussis decline with aging. The importance of tetanus–diphtheria–acellular pertussis vaccine administration should be emphasized for the protection of young adult and elderly people also, not limited to children. PMID:28168170

  7. Osteogenic function of human acellular bone loaded with bone marrow stromal cells%骨髓基质细胞复合人脱细胞骨的成骨活性

    Institute of Scientific and Technical Information of China (English)

    张旗涛; 于有; 杨林; 姚猛; 陶天遵

    2006-01-01

    BACKGROUND: To search for an alloxenogeneic bone with good load bearing function and osteoblastic activity for treating bone defects is an important study issue. We have made a comparative study on its biome chanical characteristics and found that there was no significant difference in maximum load stress, maximum pressure as compared with fresh bone of the same size. Clinicians are concerned about the osteoblastic activity and whether the osteoblastic activity can be reserved after human allogenous a cellular bone (HAB) loaded with bone marrow stromal cells (BMSCs). OBJECTIVE: To investigate the experimental effect of HAB loaded with induced BMSCs, and observe the cellular adherence and growth as well as detect its osteoblastic activity. DESIGN: Single sample experiment. SETTING: Second Affiliated Hospital of Harbin Medical University. MATERIALS: This experiment was conducted at the Experimental Center of the Second Affiliated Hospital of Harbin Medical University between January 2003 and August 2004. HAB was obtained from fresh corpse iliac bones (donated voluntarily). METHODS: Connective tissues and cell compounds of the iliac bones were removed by processing with hydroperoxide andether solution and sterilized for preparing HAB. BMSCs from living femoral shaft bone marrow were cultured immediately in ordinary and mineralized medium containing DMEM, fetal bovine serum, dexomethasone, β-glycerophophate and ascor bic acid. Proliferation and differentiation of bone stromal cells were deter mined by detecting the level of alkaline phosphatase (ALP) and osteocalcin (OCN) in the culture medium. Induced bone stromal cells solution was condensed and implanted within HAB scaffold. Cellular osteoblastic activ ity was determined through morphological observation under the light mi croscope and electron microscope as well as biochemical index detection. MAIN OUTCOME MEASURES: ① Detection results of ALP and OCN of BMSCs/HAB composite. ② Histological observation results

  8. Biomechanical properties of peripheral nerve after acellular treatment

    Institute of Scientific and Technical Information of China (English)

    MA Xin-long; SUN Xiao-lei; YANG Zhao; LI Xiu-lan; MA Jian-xiong; ZHANG Yang; YUAN Zhen-zhen

    2011-01-01

    Background Peripheral nerve injury causes a high rate of disability and a huge economic burden,and is currently one of the serious health problems in the world.The use of nerve grafts plays a vital role in repairing nerve defects.Acellular nerve grafts have been widely used in many experimental models as a peripheral nerve substitute.The purpose of this study was to test the biomechanical properties of acellular nerve grafts.Methods Thirty-four fresh sciatic nerves were obtained from 17 adult male Wistar rats (age of 3 months) and randomly assigned to 3 groups:normal control group,nerve segments underwent no treatment and were put in phosphate buffered saline (pH 7.4) and stored at 4℃ until further use; physical method group,nerve segments were frozen at -196℃ and then thawed at 37℃; and chemical method group,nerve segments were chemically extracted with the detergents Triton X-200,sulfobetaine-10 (SB-10) and sulfobetaine-16 (SB-16).After the acellularization process was completed,the structural changes of in the sciatic nerves in each group were observed by hematoxylin-eosin staining and field emission scanning electron microscopy,then biomechanical properties were tested using a mechanical apparatus (Endura TEC ELF 3200,Bose,Boston,USA).Results Hematoxylin-eosin staining and field emission scanning electron microscopy demonstrated that the effects of acellularization,demyelination,and integrity of nerve fiber tube of the chemical method were better than that of the physical method.Biomechanical testing showed that peripheral nerve grafts treated with the chemical method resulted in some decreased biomechanical properties (ultimate load,ultimate stress,ultimate strain,and mechanical work to fracture) compared with normal control nerves,but the differences were not statistically significant (P >0.05).Conclusion Nerve treated with the chemical method may be more appropriate for use in implantation than nerve treated with the physical method.

  9. Second prize: Comprehensive proteomic analysis of human calcium oxalate monohydrate kidney stone matrix.

    Science.gov (United States)

    Canales, Benjamin K; Anderson, Lorraine; Higgins, Leeann; Slaton, Joel; Roberts, Ken P; Liu, Nathan; Monga, Manoj

    2008-06-01

    Previous efforts to identify the protein content of stone matrix have been limited by the lack of technology necessary to analyze the highly insoluble protein-crystalline complex. Our study objective is to characterize the matrix of calcium oxalate monohydrate (COM) stones using a comprehensive proteomics approach. Seven pure COM stones were powdered, and proteins were extracted using four different buffer solutions. Detergent cleanup spin columns or concentrators were used to remove detergent and to exchange buffers before trypsin digestion. Tryptic peptides were analyzed with reversed-phase, high-performance liquid chromatography (RP-HPLC) and tandem mass spectrometry (MS/MS) using a QSTAR Pulsar i quadrapole time of flight mass spectrometer. Tandem mass spectra were searched against National Center for Biotechnology Information human nonredundant database using ProteinPilot 1.0 software (Applied Biosystems, Inc.) for protein hits; peptide MS/MS spectra were manually inspected. Of the four buffers, only 2% sodium dodecyl sulfate (SDS) samples had normal HPLC and MS/MS elution patterns. We identified 68 distinct proteins with 95% confidence. More than 50 of the proteins have not been previously identified in stone matrix. Of particular note, a significant number of inflammatory proteins were identified, including immunoglobulins, defensin -3, clusterin, complement C3a, kininogen, and fibrinogen. SDS reducing buffer was efficient at solubilizing proteins from stone matrix for further MS-based proteomic analysis. A variety of cellular, structural, and plasma proteins comprise COM stone matrix. Several of the stone proteins are involved in cell injury pathways, which suggests that inflammation plays a role in human COM stone formation.

  10. Nanopatterned acellular valve conduits drive the commitment of blood-derived multipotent cells

    Science.gov (United States)

    Di Liddo, Rosa; Aguiari, Paola; Barbon, Silvia; Bertalot, Thomas; Mandoli, Amit; Tasso, Alessia; Schrenk, Sandra; Iop, Laura; Gandaglia, Alessandro; Parnigotto, Pier Paolo; Conconi, Maria Teresa; Gerosa, Gino

    2016-01-01

    Considerable progress has been made in recent years toward elucidating the correlation among nanoscale topography, mechanical properties, and biological behavior of cardiac valve substitutes. Porcine TriCol scaffolds are promising valve tissue engineering matrices with demonstrated self-repopulation potentiality. In order to define an in vitro model for investigating the influence of extracellular matrix signaling on the growth pattern of colonizing blood-derived cells, we cultured circulating multipotent cells (CMC) on acellular aortic (AVL) and pulmonary (PVL) valve conduits prepared with TriCol method and under no-flow condition. Isolated by our group from Vietnamese pigs before heart valve prosthetic implantation, porcine CMC revealed high proliferative abilities, three-lineage differentiative potential, and distinct hematopoietic/endothelial and mesenchymal properties. Their interaction with valve extracellular matrix nanostructures boosted differential messenger RNA expression pattern and morphologic features on AVL compared to PVL, while promoting on both matrices the commitment to valvular and endothelial cell-like phenotypes. Based on their origin from peripheral blood, porcine CMC are hypothesized in vivo to exert a pivotal role to homeostatically replenish valve cells and contribute to hetero- or allograft colonization. Furthermore, due to their high responsivity to extracellular matrix nanostructure signaling, porcine CMC could be useful for a preliminary evaluation of heart valve prosthetic functionality. PMID:27789941

  11. Effectiveness of acellular pertussis vaccination during childhood (Spain).

    Science.gov (United States)

    Plans, P; Toledo, D; Sala, M R; Camps, N; Villanova, M; Rodríguez, R; Alvarez, J; Solano, R; García-Cenoz, M; Barrabeig, I; Godoy, P; Minguell, S

    2016-12-01

    Pertussis vaccination with 4-5 doses of acellular vaccines is recommended in Spain to all children at 2 months to 6 years of age. The effectiveness of the acellular pertussis vaccination was assessed in this study by comparing the incidence of secondary pertussis in vaccinated (4-5 doses) and unvaccinated or partially vaccinated (0-3 doses) household contacts 1-9 years old of confirmed cases of pertussis in Spain in 2012-13. Eighty-five percent of contacts had been vaccinated with 4-5 doses of acellular pertussis vaccines. During the 2-year study period, 64 cases of secondary pertussis were detected among 405 household contacts 1-9 years old: 47 among vaccinated and 17 among unvaccinated or partially vaccinated contacts. The effectiveness for preventing secondary pertussis, calculated as 1 minus the relative risk (RR) of secondary pertussis in vaccinated vs. unvaccinated/partially vaccinated contacts, was 50 % [95 % confidence interval (CI): 19-69 %, p Spain.

  12. Human versus porcine tissue sourcing for an injectable myocardial matrix hydrogel.

    Science.gov (United States)

    Johnson, Todd D; Dequach, Jessica A; Gaetani, Roberto; Ungerleider, Jessica; Elhag, Dean; Nigam, Vishal; Behfar, Atta; Christman, Karen L

    2014-01-01

    Heart failure (HF) after myocardial infarction (MI) is a leading cause of death in the western world with a critical need for new therapies. A previously developed injectable hydrogel derived from porcine myocardial matrix (PMM) has had successful results in both small and large animal MI models. In this study, we sought to evaluate the impact of tissue source on this biomaterial, specifically comparing porcine and human myocardium sources. We first developed an analogous hydrogel derived from human myocardial matrix (HMM). The biochemical and physical properties of the PMM and HMM hydrogels were then characterized, including residual dsDNA, protein content, sulfated glycosaminoglycan (sGAG) content, complex viscosity, storage and loss moduli, and nano-scale topography. Biochemical activity was investigated with in vitro studies for the proliferation of vascular cells and differentiation of human cardiomyocyte progenitor cells (hCMPCs). Next, in vivo gelation and material spread were confirmed for both PMM and HMM after intramyocardial injection. After extensive comparison, the matrices were found to be similar, yet did show some differences. Because of the rarity of collecting healthy human hearts, the increased difficulty in processing the human tissue, shifts in ECM composition due to aging, and significant patient-to-patient variability, these studies suggest that the HMM is not a viable option as a scalable product for the clinic; however, the HMM has potential as a tool for in vitro cell culture.

  13. Clinical efficacy of negative-pressure wound therapy combined with porcine acellular dermal matrix for repairing deep burn wounds in limbs%负压伤口疗法联合猪脱细胞真皮基质修复四肢深度烧伤创面的临床疗效

    Institute of Scientific and Technical Information of China (English)

    刘伟; 李峰; 陈鑫; 潘青

    2016-01-01

    Objective To observe the clinical efficacy of negative pressure wound therapy (NPWT) in combination with porcine acellular dermal matrix (ADM) dressing for repairing deep burn wounds in limbs of patients with non-surgical treatment.Methods Thirty-two patients with deep partial-thickness burn to full-thickness burn on the limbs admitted to our ward from June 2012 to December 2015,conforming to the inclusion criteria,were divided into group NPWT (n =10,treated with interval negative pressure drainage at-16.6 kPa),group ADM (n =7,treated with porcine ADM dressing),and group NPWT + ADM (n =15,treated with interval negative pressure drainage and porcine ADM dressing as above) according to the random number table and patient's consent.After being treated for 21 d,residual wounds were cured by routine dressing change using sulfadiazine silver.On post treatment day (PTD) 7,14,and 21,wound gross observation was conducted,wound drainage fluid volume was recorded,and wound healing rate was calculated.Wound secretion was collected for bacterial culture before treatment and on PTD 21,and bacterial clearance effect was recorded.The wound healing time was also recorded.Measurement data were processed with analysis of variance for repeated measurement,one-way analysis of variance,and LSD test.Eenumeration data were processed with chi-square test or Fisher's exact test.Results (1) On PTD 7,the wounds of patients in group NPWT and group NPWT + ADM were significantly shrinked as compared with those before treatment.Skin paddle scattered on the wounds of patients in group NPWT + ADM on PTD 7.The wounds of patients in group ADM were slightly shrinked on PTD 7 as compared with those before treatment.On PTD 14,the wounds of patients in group NPWT were slightly shrinked as compared with those on PTD 7,while those in group NPWT + ADM were significantly shrinked as compared with those on PTD 7.Skin paddle on the wounds of patients in group NPWT + ADM on PTD 14 were increased and fused

  14. Matrix-specified differentiation of human decidua parietalis placental stem cells.

    Science.gov (United States)

    Sridharan, Indumathi; Kim, Taeyoung; Strakova, Zuzana; Wang, Rong

    2013-08-02

    To create suitable biological scaffolds for tissue engineering and cell therapeutics, it is essential to understand the matrix-mediated specification of stem cell differentiation. To this end, we studied the effect of collagen type I on stem cell lineage specification. We altered the properties of collagen type I by incorporating carbon nanotubes (CNT). The collagen-CNT composite material was stiffer with thicker fibers and longer D-period. Human decidua parietalis stem cells (hdpPSC) were found to differentiate exclusively and rapidly towards neural cells on the collagen-CNT matrix. We attribute this accelerated neural differentiation to the enhanced structural and mechanical properties of collagen-CNT material. Strikingly, the collagen-CNT matrix, unlike collagen, imposes the neural fate by an alternate mechanism that may be independent of beta-1 integrin and beta-catenin. The study demonstrates the sensitivity of stem cells to subtle changes in the matrix and the utilization of a novel biocomposite material for efficient and directed differentiation of stem cells.

  15. Effects of enamel matrix derivative and transforming growth factor-β1 on human osteoblastic cells

    Directory of Open Access Journals (Sweden)

    Rosa Adalberto L

    2011-07-01

    Full Text Available Abstract Background Extracellular matrix proteins are key factors that influence the regenerative capacity of tissues. The objective of the present study was to evaluate the effects of enamel matrix derivative (EMD, TGF-β1, and the combination of both factors (EMD+TGF-β1 on human osteoblastic cell cultures. Methods Cells were obtained from alveolar bone of three adult patients using enzymatic digestion. Effects of EMD, TGF-β1, or a combination of both were analyzed on cell proliferation, bone sialoprotein (BSP, osteopontin (OPN and alkaline phosphatase (ALP immunodetection, total protein synthesis, ALP activity and bone-like nodule formation. Results All treatments significantly increased cell proliferation compared to the control group at 24 h and 4 days. At day 7, EMD group showed higher cell proliferation compared to TGF-β1, EMD + TGF-β1 and the control group. OPN was detected in the majority of the cells for all groups, whereas fluorescence intensities for ALP labeling were greater in the control than in treated groups; BSP was not detected in all groups. All treatments decreased ALP levels at 7 and 14 days and bone-like nodule formation at 21 days compared to the control group. Conclusions The exposure of human osteoblastic cells to EMD, TGF-β1 and the combination of factors in vitro supports the development of a less differentiated phenotype, with enhanced proliferative activity and total cell number, and reduced ALP activity levels and matrix mineralization.

  16. Assembly of fibronectin into the extracellular matrix of early and late passage human skin fibroblasts

    Energy Technology Data Exchange (ETDEWEB)

    Mann, D.M.

    1987-01-01

    The specific binding of soluble /sup 125/I-human plasma fibronectin (/sup 125/I-HFN-P) to confluent cultures of early and late passage human skin fibroblasts was investigated. Previous studies HFN-P bound to fibroblast cell layers indicated that HNF-P was present in the cultures in two separate pools, distinguishable on the basis of their solubility in 1% deoxycholate. Examination of the kinetics of /sup 125/I-HFN-P binding to Pool I of early and late passage cultures revealed that both cultures required 2-4 h to approach steady-state conditions. Other kinetic studies showed that the rates of low of /sup 125/I-HFN-P from either Pool I or Pool II were similar for both cultures. Further, Scatchard analysis revealed a single class of Pool I binding sites with apparent dissociation constants (K/sub d/) of 5.3 x 10/sup -8/M (early passage) and 4.2 x 10/sup -8/M (late passage). These results indicate that early and late passage cultures of human fibroblasts exhibit differences in the number of cell surface biding sites for soluble fibronectin, and in the extent to which they incorporate soluble fibronectin into the extracellular matrix. Parameters which affect the fibronectin matrix assembly system of human skin fibroblasts were also examined. In addition, several monoclonal anti-fibronectin antibodies were characterized and developed as experimental probes for fibronectin structure and function.

  17. Acellular dermal matrix for repair of porcine bile duct defects:to promote vascular and bile duct epithelial regeneration%脱细胞真皮基质修复猪胆管缺损:促进血管及胆管上皮再生

    Institute of Scientific and Technical Information of China (English)

    陈刚; 白建华; 朱新锋; 曹俊; 刘其雨; 赵英鹏; 李立

    2015-01-01

    BACKGROUND:Acelular dermal matrix is a cel-free natural tissue scaffold similar to human soft tissue, which is easy to shape and has non-toxic side effects. It has been used to repair the urethra and ureter. OBJECTIVE:To investigate the effect of acelular dermal matrix on the repair of bile duct injury. METHODS:Thirty Diannan miniature pigs were randomly divided into three groups: in blank group, the bile duct was resected folowed by end to end anastomosis; in experimental group, bile duct defect model was made folowed by repair with acelular dermal matrix; in control group, bile duct defect model was made folowed by repair with expanded polytetrafluoroethylene. At 6 and 24 weeks after repair, bile duct patches and surrounding tissues were taken for immunohistochemical observation and RT-PCR detection. RESULTS AND CONCLUSION: Compared with the control and blank group, the expression of cytokeratin was higher, but the expression of transforming growth factor β1 was lower in the experimental group. Within 24 weeks after repair, the total mRNA level of transforming growth factor β1 was lower in the experimental group than the other two groups (P < 0.05), but the total mRNA levels of insulin-like growth factor 2 and vascular endothelial growth factor were higher in the experimental group (P < 0.05). These findings indicate that the acelular dermal matrix for repair of bile duct injury can promote angiogenesis and bile duct epithelial regeneration, but not increase the formation of scars.%背景:脱细胞真皮基质是无细胞的天然组织支架,与人体软组织十分相近,易于塑形,无毒副作用,已被用于修补尿道与输尿管。目的:观察脱细胞基质修补胆管损伤的效果。方法:将30头滇南小耳猪随机均分为3组,空白对照组切断胆管后行端端吻合,实验组人为制作胆管缺损后以脱细胞真皮基质修补,对照组人为制作胆管缺损后以膨体聚四氟乙烯修补。修补后6,24

  18. Food Matrix Effects of Polyphenol Bioaccessibility from Almond Skin during Simulated Human Digestion

    Science.gov (United States)

    Mandalari, Giuseppina; Vardakou, Maria; Faulks, Richard; Bisignano, Carlo; Martorana, Maria; Smeriglio, Antonella; Trombetta, Domenico

    2016-01-01

    The goal of the present study was to quantify the rate and extent of polyphenols released in the gastrointestinal tract (GIT) from natural (NS) and blanched (BS) almond skins. A dynamic gastric model of digestion which provides a realistic simulation of the human stomach was used. In order to establish the effect of a food matrix on polyphenols bioaccessibility, NS and BS were either digested in water (WT) or incorporated into home-made biscuits (HB), crisp-bread (CB) and full-fat milk (FM). Phenolic acids were the most bioaccessible class (68.5% release from NS and 64.7% from BS). WT increased the release of flavan-3-ols (p antioxidant status in the digestion medium, indicating that phenolic compounds could bind protein present in the food matrix. The release of bioactives from almond skins could explain the beneficial effects associated with almond consumption. PMID:27649239

  19. Protein crystallization with microseed matrix screening: application to human germline antibody Fabs

    Energy Technology Data Exchange (ETDEWEB)

    Obmolova, Galina, E-mail: gobmolov@its.jnj.com; Malia, Thomas J.; Teplyakov, Alexey; Sweet, Raymond W.; Gilliland, Gary L., E-mail: gobmolov@its.jnj.com [Janssen Research and Development LLC, 1400 McKean Road, Spring House, PA 19477 (United States)

    2014-07-23

    The power of microseed matrix screening is demonstrated in the crystallization of a panel of antibody Fab fragments. The crystallization of 16 human antibody Fab fragments constructed from all pairs of four different heavy chains and four different light chains was enabled by employing microseed matrix screening (MMS). In initial screening, diffraction-quality crystals were obtained for only three Fabs, while many Fabs produced hits that required optimization. Application of MMS, using the initial screens and/or refinement screens, resulted in diffraction-quality crystals of these Fabs. Five Fabs that failed to give hits in the initial screen were crystallized by cross-seeding MMS followed by MMS optimization. The crystallization protocols and strategies that resulted in structure determination of all 16 Fabs are presented. These results illustrate the power of MMS and provide a basis for developing future strategies for macromolecular crystallization.

  20. Aggrecan-based extracellular matrix is an integral part of the human basal ganglia circuit.

    Science.gov (United States)

    Brückner, G; Morawski, M; Arendt, T

    2008-01-24

    The extracellular matrix is known to be involved in neuronal communication and the regulation of plastic changes, and also considered to protect neurons and synapses against damage. The goal of this study was to investigate how major extracellular matrix components (aggrecan, link protein, hyaluronan) constitute the pathways of the nigral system in the human basal ganglia circuit affected by neurodegeneration in Parkinson's disease. Here we show that aggrecan- and link protein-related components form clear regional distribution patterns, whereas hyaluronan is widely distributed in gray and white matter. Two predominant phenotypes of the aggrecan-based matrix can be discriminated: (1) perineuronal nets (PNs) and (2) axonal coats (ACs) encapsulating preterminal fibers and synaptic boutons. Clearly contoured PNs are associated with GABAergic projection neurons in the external and internal division of the globus pallidus, the lateral and reticular part of the substantia nigra, as well as subpopulations of striatal and thalamic inhibitory interneurons. Dopaminergic nigral neurons are devoid of PNs but are contacted to a different extent by matrix-coated boutons forming subnucleus-specific patterns. A very dense network of ACs is characteristic especially of the posterior lateral cell groups of the compact substantia nigra (nigrosome 1). In the subthalamic nucleus and the lateral thalamic nuclei numerous AC-associated axons were attached to principal neurons devoid of PNs. We conclude from the region-specific patterns that the aggrecan-based extracellular matrix is adapted to the fast processing of sensorimotor activities which are the therapeutic target of surgery and deep brain stimulation in the treatment of advanced stages of Parkinson's disease.

  1. Establishment and characterization of human engineered cells stably expressing large extracellular matrix proteins.

    Science.gov (United States)

    Kwon, Daekee; Kang, Gwang-Sik; Han, Dong Keun; Park, Kwideok; Kim, Jae-Hwan; Lee, Soo-Hong

    2014-01-01

    Commercially available extracellular matrix (ECM) hydrogel-coated culture plates have been used to study the relationship between the ECM microenvironment and stem cell behavior. However, it is unclear whether ECM-coated dishes mimic the natural ECM microenvironment because the architecture of the ECM is constructed of randomly distributed fibers. The purpose of this study was the production and confirmation of human engineered cell lines stably expressing large ECM proteins such as collagen I/II and fibronectin. First, large (over 10 kb) ECM vectors encoding human collagen I/II and fibronectin were constructed and the circular vectors were linearized. Second, the linear ECM vectors were introduced into immortalized human embryonic kidney cells using various transfection methods. The polyethylenimine and liposome methods showed higher efficiencies than electroporation for transfection of these large vectors. Third, human ECM engineered cells were established by stable integration of the vector into the genomic DNA and resulted in stable overexpression of mRNA and proteins. In summary, human engineered cell lines stably expressing large ECM proteins such as human collagen I/II and fibronectin were successfully prepared, and secretion of the ECM components into the surrounding environment was confirmed by immunocytochemistry. Thus, human ECM engineered cells naturally secreting ECM components could be valuable for studying the relationship between the native ECM microenvironment and stem cell behavior.

  2. Decellularized matrix from tumorigenic human mesenchymal stem cells promotes neovascularization with galectin-1 dependent endothelial interaction.

    Directory of Open Access Journals (Sweden)

    Jorge S Burns

    Full Text Available BACKGROUND: Acquisition of a blood supply is fundamental for extensive tumor growth. We recently described vascular heterogeneity in tumours derived from cell clones of a human mesenchymal stem cell (hMSC strain (hMSC-TERT20 immortalized by retroviral vector mediated human telomerase (hTERT gene expression. Histological analysis showed that cells of the most vascularized tumorigenic clone, -BD11 had a pericyte-like alpha smooth muscle actin (ASMA+ and CD146+ positive phenotype. Upon serum withdrawal in culture, -BD11 cells formed cord-like structures mimicking capillary morphogenesis. In contrast, cells of the poorly tumorigenic clone, -BC8 did not stain for ASMA, tumours were less vascularized and serum withdrawal in culture led to cell death. By exploring the heterogeneity in hMSC-TERT20 clones we aimed to understand molecular mechanisms by which mesenchymal stem cells may promote neovascularization. METHODOLOGY/PRINCIPAL FINDINGS: Quantitative qRT-PCR analysis revealed similar mRNA levels for genes encoding the angiogenic cytokines VEGF and Angiopoietin-1 in both clones. However, clone-BD11 produced a denser extracellular matrix that supported stable ex vivo capillary morphogenesis of human endothelial cells and promoted in vivo neovascularization. Proteomic characterization of the -BD11 decellularized matrix identified 50 extracellular angiogenic proteins, including galectin-1. siRNA knock down of galectin-1 expression abrogated the ex vivo interaction between decellularized -BD11 matrix and endothelial cells. More stable shRNA knock down of galectin-1 expression did not prevent -BD11 tumorigenesis, but greatly reduced endothelial migration into -BD11 cell xenografts. CONCLUSIONS: Decellularized hMSC matrix had significant angiogenic potential with at least 50 angiogenic cell surface and extracellular proteins, implicated in attracting endothelial cells, their adhesion and activation to form tubular structures. hMSC -BD11 surface galectin-1

  3. Alteration of nuclear matrix-intermediate filament system and differential expression of nuclear matrix proteins during human hepatocarcinoma cell differentiation

    Institute of Scientific and Technical Information of China (English)

    Jian Tang; Jing-Wen Niu; Dong-Hui Xu; Zhi-Xing Li; Qi-Fu Li; Jin-An Chen

    2007-01-01

    AIM:To investigate the association between the configurational and compositional changes of nuclear matrix and the differentiation of carcinoma cells.METHODS: Cells cultured with or without 5 × 10-3mmol/L of hexamethylene bisacetamide (HMBA) on Nickel grids were treated by selective extraction and prepared for whole mount observation under electron microscopy. The samples were examined under transmission electron microscope. Nuclear matrix proteins were selectively extracted and subjected to subcellular proteomics study. The protein expression patterns were analyzed by PDQuest software. Spots of differentially expressed nuclear matrix proteins were excised and subjected to in situ digestion with trypsin.The peptides were analyzed by matrix-assisted laserdesorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). Data were submitted for database searching using Mascot tool (www. Matrixscience.com).RESULTS: The nuclear matrix (NM) and intermediate filament (IF) in SMMC-7721 hepatocarcinoma cells were found relatively sparse and arranged irregularly.The nuclear lamina was non-uniform, and two kinds of filaments were not tightly connected. After induction for differentiation by HMBA, the NM-IF filaments were concentrated and distributed uniformly. The heterogeneous population of filaments, including highly branched utrathin filaments could also be seen in the regular meshwork. The connection between the two kinds of filaments and the relatively thin, condensed and sharply demarcated lamina composed of intermediatesized filaments was relatively fastened. Meanwhile, 21NM proteins changed remarkably during SMMC-7721cell differentiation. Four proteins, I.e. Mutant Pyst1,hypothetical protein, nucleophosmin1, and LBP were downregulated, whereas four other proteins, eIF6, p44subunit, β-tubulin, and SIN3B were upregulated with the last one, SR2/ASF found only in the differentiated SMMC-7721 cells.CONCLUSION: The induced differentiation of SMMC-7721cells by HMBA is

  4. The parasite Entamoeba histolytica exploits the activities of human matrix metalloproteinases to invade colonic tissue.

    Science.gov (United States)

    Thibeaux, Roman; Avé, Patrick; Bernier, Michèle; Morcelet, Marie; Frileux, Pascal; Guillén, Nancy; Labruyère, Elisabeth

    2014-10-07

    Intestinal invasion by the protozoan parasite Entamoeba histolytica is characterized by remodelling of the extracellular matrix (ECM). The parasite cysteine proteinase A5 (CP-A5) is thought to cooperate with human matrix metalloproteinases (MMPs) involved in ECM degradation. Here, we investigate the role CP-A5 plays in the regulation of MMPs upon mucosal invasion. We use human colon explants to determine whether CP-A5 activates human MMPs. Inhibition of the MMPs' proteolytic activities abolishes remodelling of the fibrillar collagen structure and prevents trophozoite invasion of the mucosa. In the presence of trophozoites, MMPs-1 and -3 are overexpressed and are associated with fibrillar collagen remodelling. In vitro, CP-A5 performs the catalytic cleavage needed to activate pro-MMP-3, which in turn activates pro-MMP-1. Ex vivo, incubation with recombinant CP-A5 was enough to rescue CP-A5-defective trophozoites. Our results suggest that MMP-3 and/or CP-A5 inhibitors may be of value in further studies aiming to treat intestinal amoebiasis.

  5. Comprehensive analysis of leukocytes, vascularization and matrix metalloproteinases in human menstrual xenograft model.

    Science.gov (United States)

    Guo, Yong; He, Bin; Xu, Xiangbo; Wang, Jiedong

    2011-02-17

    In our previous study, menstrual-like changes in mouse were provoked through the pharmacologic withdrawal of progesterone with mifepristone following induction of decidualization. However, mouse is not a natural menstruation animal, and the menstruation model using external stimuli may not truly reflect the occurrence and development of the human menstrual process. Therefore, we established a model of menstruation based on human endometrial xenotransplantation. In this model, human endometrial tissues were transplanted subcutaneously into SCID mice that were ovarectomized and supplemented with estrogen and progestogen by silastic implants with a scheme imitating the endocrinological milieu of human menstrual cycle. Morphology, hormone levels, and expression of vimentin and cytokeratin markers were evaluated to confirm the menstrual-like changes in this model. With 28 days of hormone treatment, transplanted human endometrium survived and underwent proliferation, differentiation and disintegration, similar to human endometrium in vivo. Human CD45+ cells showed a peak of increase 28 days post-transplantation. Three days after progesterone withdrawal, mouse CD45+ cells increased rapidly in number and were significantly greater than human CD45+ cell counts. Mouse CD31+ blood vascular-like structures were detected in both transplanted and host tissues. After progesterone withdrawal, the expression levels of matrix metalloproteinases (MMP) 1, 2, and 9 were increased. In summary, we successfully established a human endometrial xenotransplantation model in SCID mice, based on the results of menstrual-like changes in which MMP-1, 2 and 9 are involved. We showed that leukocytes are originated from in situ proliferation in human xenografts and involved in the occurrence of menstruation. This model will help to further understand the occurrence, growth, and differentiation of the endometrium and the underlying mechanisms of menstruation.

  6. Comprehensive analysis of leukocytes, vascularization and matrix metalloproteinases in human menstrual xenograft model.

    Directory of Open Access Journals (Sweden)

    Yong Guo

    Full Text Available In our previous study, menstrual-like changes in mouse were provoked through the pharmacologic withdrawal of progesterone with mifepristone following induction of decidualization. However, mouse is not a natural menstruation animal, and the menstruation model using external stimuli may not truly reflect the occurrence and development of the human menstrual process. Therefore, we established a model of menstruation based on human endometrial xenotransplantation. In this model, human endometrial tissues were transplanted subcutaneously into SCID mice that were ovarectomized and supplemented with estrogen and progestogen by silastic implants with a scheme imitating the endocrinological milieu of human menstrual cycle. Morphology, hormone levels, and expression of vimentin and cytokeratin markers were evaluated to confirm the menstrual-like changes in this model. With 28 days of hormone treatment, transplanted human endometrium survived and underwent proliferation, differentiation and disintegration, similar to human endometrium in vivo. Human CD45+ cells showed a peak of increase 28 days post-transplantation. Three days after progesterone withdrawal, mouse CD45+ cells increased rapidly in number and were significantly greater than human CD45+ cell counts. Mouse CD31+ blood vascular-like structures were detected in both transplanted and host tissues. After progesterone withdrawal, the expression levels of matrix metalloproteinases (MMP 1, 2, and 9 were increased. In summary, we successfully established a human endometrial xenotransplantation model in SCID mice, based on the results of menstrual-like changes in which MMP-1, 2 and 9 are involved. We showed that leukocytes are originated from in situ proliferation in human xenografts and involved in the occurrence of menstruation. This model will help to further understand the occurrence, growth, and differentiation of the endometrium and the underlying mechanisms of menstruation.

  7. Tissue-engineered acellular matrix material:preparation and application in articular cartilage repair%脱细胞基质材料制备方法及在骨关节软骨损伤修复中的应用

    Institute of Scientific and Technical Information of China (English)

    赵玉果; 李明明

    2016-01-01

      结果与结论:①脱细胞基质组织工程材料交联后呈现为深蓝色,疏松多孔,直径为5 mm,硬度适中,具备一定的弹性;②苏木精-伊红染色不含有细胞碎屑及蓝染的核物质,不存在残留的细胞外基质;③甲苯胺蓝染色为蓝色材料支架孔隙率为90%,溶胀率为(1314±337)%;④脱细胞基质组织组材料1,3,5,7,9 d的A值显著高于纤维样组织组(P OBJECTIVE:To investigate the effect of tissue-engineered acelular matrix in articular cartilage repair. METHODS:Totaly 30 New Zealand rabbits were randomly alottedto fibroid tissue andacelular matrix groups (n=15 per group), and then articular cartilage defect models,4mmin diameter,were established at the white rabbitfemoral condyle. Acelular cartilage matrix scaffold was prepared using bovine knee cartilage, and model rats in the acelular matrix group were repaired with acelular cartilage matrix scaffold and the others in the fibroid tissue group repaired with fibroid tissues. Finaly, repair effects between two groups were compared. RESULTS AND CONCLUSION:The dark blue and porous tissue-engineered acelular matrix material could be found, with a diameter of 5mm and moderate hardness, and exhibited certain flexibility after cross-linking. Hematoxylin-eosin staining showed that cel debris,blue-stainednuclear materials and residual extracelular matrix disappeared. Toluidine blue staining found that the porosity of the blue scaffold was 90%, and the sweling ratio was (1314±337)%. The absorbance value in the acelular matrix group was significantly higher than that in the fibroid tissue group at 1, 3, 5, 7 and 9 days (P< 0.05). In the fibroid tissue group,defectsfiled withnewborn fibrous scars were overt. By contrast, in the acelularmatrix group, the white tissuescovered the defect regionwith smooth surface,and the woundwas basicaly healed,withanunclearboundaryafter 12weeks. Moreover, blue-stained, smal flattened cels appeared

  8. Chemically extracted acellular allogeneic nerve graft combined with ciliary neurotrophic factor promotes sciatic nerve repair

    Institute of Scientific and Technical Information of China (English)

    Yanru Zhang; Hui Zhang; Kaka Katiella; Wenhua Huang

    2014-01-01

    A chemically extracted acellular allogeneic nerve graft can reduce postoperative immune re-jection, similar to an autologous nerve graft, and can guide neural regeneration. However, it remains poorly understood whether a chemically extracted acellular allogeneic nerve graft combined with neurotrophic factors provides a good local environment for neural regenera-tion. This study investigated the repair of injured rat sciatic nerve using a chemically extracted acellular allogeneic nerve graft combined with ciliary neurotrophic factor. An autologous nerve anastomosis group and a chemical acellular allogeneic nerve bridging group were prepared as controls. At 8 weeks after repair, sciatic functional index, evoked potential amplitude of the soleus muscle, triceps wet weight recovery rate, total number of myelinated nerve fibers and myelin sheath thickness were measured. For these indices, values in the three groups showed the autologous nerve anastomosis group > chemically extracted acellular nerve graft + ciliary neurotrophic factor group > chemical acellular allogeneic nerve bridging group. These results suggest that chemically extracted acellular nerve grafts combined with ciliary neurotrophic factor can repair sciatic nerve defects, and that this repair is inferior to autologous nerve anasto-mosis, but superior to chemically extracted acellular allogeneic nerve bridging alone.

  9. RGD-Dependent Epithelial Cell-Matrix Interactions in the Human Intestinal Crypt

    Directory of Open Access Journals (Sweden)

    Yannick D. Benoit

    2012-01-01

    Full Text Available Interactions between the extracellular matrix (ECM and integrin receptors trigger structural and functional bonds between the cell microenvironment and the cytoskeleton. Such connections are essential for adhesion structure integrity and are key players in regulating transduction of specific intracellular signals, which in turn regulate the organization of the cell microenvironment and, consequently, cell function. The RGD peptide-dependent integrins represent a key subgroup of ECM receptors involved in the maintenance of epithelial homeostasis. Here we review recent findings on RGD-dependent ECM-integrin interactions and their roles in human intestinal epithelial crypt cells.

  10. Optimization of Substitution Matrix for Sequence Alignment of Major Capsid Proteins of Human Herpes Simplex Virus

    Directory of Open Access Journals (Sweden)

    Vipan Kumar Sohpal

    2011-12-01

    Full Text Available Protein sequence alignment has become an informative tool in modern molecular biology research. A number of substitution matrices have been readily available for sequence alignments, but it is challenging task to compute optimal matrices for alignment accuracy. Here, we used the parameter optimization procedure to select the optimal Q of substitution matrices for major viral capsid protein of human herpes simplex virus. Results predict that Blosum matrix is most accurate on alignment benchmarks, and Blosum 60 provides the optimal Q in all substitution matrices. PAM 200 matrices results slightly below than Blosum 60, while VTML matrices are intermediate of PAM and VT matrices under dynamic programming.

  11. Matrix metalloproteinase sensing via porous silicon microcavity devices functionalized with human antibodies

    Energy Technology Data Exchange (ETDEWEB)

    Martin, Marta; Gergely, Csilla [GES-UMR 5650, CNRS, Universite Montpellier 2, Pl. Eugene Bataillon 34095, Montpellier Cedex 5 (France); Taleb Bendiab, Chakib; Massif, Laurent; Cuisinier, Frederic [EA4203, Faculte d' Odontologie, Universite Montpellier 1, Montpellier Cedex 5 (France); Palestino, Gabriela [Facultad de Ciencias Quimicas, Universidad Autonoma de San Luis Potosi, Av. Salvador Nava 6, 78000 San Luis Potosi (Mexico); Agarwal, Vivechana [CIICAP, Universidad Autonoma del Estado de Morelos, Av. Universidad 1001, Col Chamilpa, Cuernavaca, Mor. (Mexico)

    2011-06-15

    Porous silicon microcavity (PSiMc) structures were used as support material for specific sensing of matrix metalloproteinases (MMPs). For lower concentrations of MMP-8, the structures were tested with two types of functionalization methods. Silanization of the oxidized porous silicon structures, followed by glutaraldehyde chemistry was found to give very inconsistent results. The use of biotinilated bovine serum albumin linked to the naked PSiMc was found to be an alternative method to attach the anti MMP-8 human antibody, previously modified with streptavidin, which was further used to sense MMP-8 (copyright 2011 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim) (orig.)

  12. Human hamstring tenocytes survive when seeded into a decellularized porcine Achilles tendon extracellular matrix.

    Science.gov (United States)

    Lohan, Anke; Stoll, Christiane; Albrecht, Marit; Denner, Andreas; John, Thilo; Krüger, Kay; Ertel, Wolfgang; Schulze-Tanzil, Gundula

    2013-01-01

    Tendon ruptures and defects remain major orthopaedic challenges. Tendon healing is a time-consuming process, which results in scar tissue with an altered biomechanical competence. Using a xenogeneic tendon extracellular matrix (ECM) as a natural scaffold, which can be reseeded with autologous human tenocytes, might be a promising approach to reconstruct damaged tendons. For this purpose, the porcine Achilles (AS) tendons serving as a scaffold were histologically characterized in comparison to human cell donor tendons. AS tendons were decellularized and then reseeded with primary human hamstring tenocytes using cell centrifuging, rotating culture and cell injection techniques. Vitality testing, histology and glycosaminoglycan/DNA quantifications were performed to document the success of tendon reseeding. Porcine AS tendons were characterized by a higher cell and sulfated glycosaminoglycan content than human cell donor tendons. Complete decellularization could be achieved, but led to a wash out of sulfated glycosaminoglycans. Nevertheless, porcine tendon could be recellularized with vital human tenocytes. The recellularization led to a slight increase in cell number compared to the native tendon and some glycosaminoglycan recovery. This study indicates that porcine tendon can be de- and recellularized using adult human tenocytes. Future work should optimize cell distribution within the recellularized tendon ECM and consider tendon- and donor species-dependent differences.

  13. EFFECTS OF GENISTEIN ON INVASION AND MATRIX METALLOPROTEINASE ACTIVITIES OF HT1080 HUMAN FIBROSARCOMA CELLS

    Institute of Scientific and Technical Information of China (English)

    1999-01-01

    @@ Effects of genistein on invasion and matrix metalloproteinase activities were investigated in HT1080 human sarcoma cells.Invasion of HT1080 cells through reconstituted basement membrane was inhibited when the cells were treated with 100 μ mol/L and 200 μ mol/L genistein.At the same concentrations,genistein not only suppressed latent forms of matrix metalloprotinese-2 and-9(MMP-2 and MMP-9) to convert into active forms,but also increase dramatically the tissue inhibitor of metalloproteinase(TIMP-1) mRNA contents and reverse the imbalance of MMPs and TIMPs.However,expressions of MMP-2 and MMP-9 were not significantly affected.Suppression of MMP activation and increase of TIMP-1 expression will decrease matrix degradation by MMPs,and consequently inhibit invasions of the cells.These results emphasized the existence of the imbalance between MMPs and TIMPs in tumor invasion and metastasis formation.The value of genistein as a drug for antiinvasion and anti-metastasis chemotherapy was suggested.

  14. EFFECTS OF GENISTEIN ON INVASION AND MATRIX METALLOPROTEINASE ACTIVITIES OF HT1080 HUMAN FIBROSARCOMA CELLS

    Institute of Scientific and Technical Information of China (English)

    颜春洪; 韩锐

    1999-01-01

    Effects of genistein on invasion and matrix metalloproteinase activities were investigated in HT1080 human sarcoma cells, lnvasion of HTI080 cells through reconstituted basement membnme was inhibited when the cells were treated with 100μmol/L and 200μmol/L genistein; At the same concentrations,genistein not only suppressed latent forms of matrix metalloprotinese-2 and-9 (MMP-2 mad MMP-9) to convert into active forms, but also increase dramatically the tissue inhibitor of metalloproteinase (TIMP-1 ) mRNA contents and reverse the imbalance of MMPs and TIMPs. However, expressions of MMP-2 and MMP-9 were not sigrdficantly affected. Suppression of MMP activation and increase of TMP-1 expression will decrease matrix degradation by MMPs, and consequently inhibit invasions of the cells. These results emphasized the existence of the imbalance between MMPs and TIMPs in tumor invasion mad metastasis formation, The value of genistein as a drug for antiinvasion and anti-metastasis chemotherapy was suggested.

  15. Muscarinic receptor agonists stimulate matrix metalloproteinase 1-dependent invasion of human colon cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Raufman, Jean-Pierre, E-mail: jraufman@medicine.umaryland.edu [Division of Gastroenterology and Hepatology, University of Maryland School of Medicine, Baltimore, MD (United States); Cheng, Kunrong; Saxena, Neeraj; Chahdi, Ahmed; Belo, Angelica; Khurana, Sandeep; Xie, Guofeng [Division of Gastroenterology and Hepatology, University of Maryland School of Medicine, Baltimore, MD (United States)

    2011-11-18

    Highlights: Black-Right-Pointing-Pointer Muscarinic receptor agonists stimulated robust human colon cancer cell invasion. Black-Right-Pointing-Pointer Anti-matrix metalloproteinase1 antibody pre-treatment blocks cell invasion. Black-Right-Pointing-Pointer Bile acids stimulate MMP1 expression, cell migration and MMP1-dependent invasion. -- Abstract: Mammalian matrix metalloproteinases (MMPs) which degrade extracellular matrix facilitate colon cancer cell invasion into the bloodstream and extra-colonic tissues; in particular, MMP1 expression correlates strongly with advanced colon cancer stage, hematogenous metastasis and poor prognosis. Likewise, muscarinic receptor signaling plays an important role in colon cancer; muscarinic receptors are over-expressed in colon cancer compared to normal colon epithelial cells. Muscarinic receptor activation stimulates proliferation, migration and invasion of human colon cancer cells. In mouse intestinal neoplasia models genetic ablation of muscarinic receptors attenuates carcinogenesis. In the present work, we sought to link these observations by showing that MMP1 expression and activation plays a mechanistic role in muscarinic receptor agonist-induced colon cancer cell invasion. We show that acetylcholine, which robustly increases MMP1 expression, stimulates invasion of HT29 and H508 human colon cancer cells into human umbilical vein endothelial cell monolayers - this was abolished by pre-incubation with atropine, a non-selective muscarinic receptor inhibitor, and by pre-incubation with anti-MMP1 neutralizing antibody. Similar results were obtained using a Matrigel chamber assay and deoxycholyltaurine (DCT), an amidated dihydroxy bile acid associated with colon neoplasia in animal models and humans, and previously shown to interact functionally with muscarinic receptors. DCT treatment of human colon cancer cells resulted in time-dependent, 10-fold increased MMP1 expression, and DCT-induced cell invasion was also blocked by pre

  16. Nicotine Treatment Induces Expression of Matrix Metalloproteinases in Human Osteoblastic Saos-2 Cells

    Institute of Scientific and Technical Information of China (English)

    Tomoko KATONO; Takayuki KAWATO; Natsuko TANABE; Naoto SUZUKI; Kazuhiro YAMANAKA; Hitoshi OKA; Masafumi MOTOHASHI; Masao MAENO

    2006-01-01

    Tobacco smoking is an important risk factor for the development of severe periodontitis.Recently, we showed that nicotine affected mineralized nodule formation, and that nicotine and lipopolysaccharide stimulated the formation of osteoclast-like cells by increasing production of macrophage colony-stimulating factor (M-CSF) and prostaglandin E2 (PGE2) by human osteoblastic Saos-2 cells. In the present study, we examined the effects of nicotine on the expression of matrix metalloproteinases (MMPs),tissue inhibitors of matrix metalloproteinases (TIMPs), the plasminogen activation system including the component of tissue-type plasminogen activator (tPA), urokinase-type PA (uPA), and PA inhibitor type 1(PAI- 1), α7 nicotine receptor, and c-fos. We also examined the effect of the nicotine antagonist D-tubocurarine on nicotine-induced expression of MMP-1. Gene expression was examined using real-time polymerase chain reaction (PCR) to estimate mRNA levels. In addition, expression of the MMP, TIMP, uPA, tPA, and PAI-1proteins was determined by Western blotting analysis. Nicotine treatment caused expression of MMP-1, 2, 3,and 13, but not MMP-14, to increase significantly after 5 or 10 d of culture; MMP-14 expression did not change through day 14. Enhancement of MMP-1 expression by nicotine treatment was eliminated by simultaneous treatment with D-tubocurarine. In the presence of nicotine, expression of uPA, PAI-1, or TIMP-1, 2, 3, or 4 did not change over 14 d of culture, whereas expression of tPA increased significantly by day 7. Nicotine also increased expression of the α7 nicotine receptor and c-fos genes. These results suggest that nicotine stimulates bone matrix turnover by increasing production of tPA and MMP-1, 2, 3, and 13,thereby tipping the balance between bone matrix formation and resorption toward the latter process.

  17. Identification of Extracellular Matrix Components and Biological Factors in Micronized Dehydrated Human Amnion/Chorion Membrane

    Science.gov (United States)

    Lei, Jennifer; Priddy, Lauren B.; Lim, Jeremy J.; Massee, Michelle; Koob, Thomas J.

    2017-01-01

    Objective: The use of bioactive extracellular matrix (ECM) grafts such as amniotic membranes is an attractive treatment option for enhancing wound repair. In this study, the concentrations, activity, and distribution of matrix components, growth factors, proteases, and inhibitors were evaluated in PURION® Processed, micronized, dehydrated human amnion/chorion membrane (dHACM; MiMedx Group, Inc.). Approach: ECM components in dHACM tissue were assessed by using immunohistochemical staining, and growth factors, cytokines, proteases, and inhibitors were quantified by using single and multiplex ELISAs. The activities of proteases that were native to the tissue were determined via gelatin zymography and EnzChek® activity assay. Results: dHACM tissue contained the ECM components collagens I and IV, hyaluronic acid, heparin sulfate proteoglycans, fibronectin, and laminin. In addition, numerous growth factors, cytokines, chemokines, proteases, and protease inhibitors that are known to play a role in the wound-healing process were quantified in dHACM. Though matrix metalloproteinases (MMPs) were present in dHACM tissues, inhibitors of MMPs overwhelmingly outnumbered the MMP enzymes by an overall molar ratio of 28:1. Protease activity assays revealed that the MMPs in the tissue existed primarily either in their latent form or complexed with inhibitors. Innovation: This is the first study to characterize components that function in wound healing, including inhibitor and protease content and activity, in micronized dHACM. Conclusion: A variety of matrix components and growth factors, as well as proteases and their inhibitors, were identified in micronized dHACM, providing a better understanding of how micronized dHACM tissue can be used to effectively promote wound repair. PMID:28224047

  18. Whooping cough, twenty years from acellular vaccines introduction.

    Science.gov (United States)

    Greco, D; Esposito, S; Tozzi, A; Pandolfi, E; Icardi, G; Giammanco, A

    2015-01-01

    Clinical pertussis resulting from infection with B. pertussis is a significant medical and public health problem, despite the huge success of vaccination that has greatly reduced its incidence. The whole cell vaccine had an undeniable success over the last 50 years, but its acceptance was strongly inhibited by fear, only partially justified, of severe side effects, but also, in the Western world, by the difficulty to enter in combination with other vaccines: today multi-vaccine formulations are essential to maintain a high vaccination coverage. The advent of acellular vaccines was greeted with enthusiasm by the public health world: in the Nineties, several controlled vaccine trials were carried out: they demonstrated a high safety and good efficacy of new vaccines. In fact, in the Western world, the acellular vaccines completely replaced the whole cells ones. In the last years, ample evidence on the variety of protection of these vaccines linked to the presence of different antigens of Bordetella pertussis was collected. It also became clear that the protection provided, on average around 80%, leaves every year a significant cohort of vaccinated susceptible even in countries with a vaccination coverage of 95%, such as Italy. Finally, it was shown that, as for the pertussis disease, protection decreases over time, to leave a proportion of adolescents and adults unprotected. Waiting for improved pertussis vaccines, the disease control today requires a different strategy that includes a booster at 5 years for infants, but also boosters for teenagers and young adults, re-vaccination of health care personnel, and possibly of pregnant women and of those who are in contact with infants (cocooning). Finally, the quest for better vaccines inevitably tends towards pertussis acellular vaccines with at least three components, which have demonstrated superior effectiveness and have been largely in use in Italy for fifteen years.

  19. Long-Term Followup of Dermal Substitution with Acellular Dermal Implant in Burns and Postburn Scar Corrections

    Directory of Open Access Journals (Sweden)

    I. Juhasz

    2010-01-01

    Full Text Available Full-thickness burn and other types of deep skin loss will result in scar formation. For at least partial replacement of the lost dermal layer, there are several options to use biotechnologically derived extracellular matrix components or tissue scaffolds of cadaver skin origin. In a survey, we have collected data on 18 pts who have previously received acellular dermal implant Alloderm. The age of these patients at the injury varied between 16 months and 84 years. The average area of the implants was 185 cm2. Among those, 15 implant sites of 14 patients were assessed at an average of 50 months after surgery. The scar function was assessed by using the modified Vancouver Scar Scale. We have found that the overall scar quality and function was significantly better over the implanted areas than over the surrounding skin. Also these areas received a better score for scar height and pliability. Our findings suggest that acellular dermal implants are especially useful tools in the treatment of full-thickness burns as well as postburn scar contractures.

  20. Development and preclinical evaluation of acellular collagen scaffolding and autologous artificial connective tissue in the regeneration of oral mucosa wounds.

    Science.gov (United States)

    Espinosa, Lady; Sosnik, Alejandro; Fontanilla, Marta R

    2010-05-01

    This work assessed wound healing response in rabbit oral lesions grafted with autologous artificial connective tissue or acellular collagen scaffolds. Autologous artificial oral connective tissue (AACT) was produced using rabbit fibroblasts and collagen I scaffolds. Before implantation, AACT grafts were assayed to demonstrate the presence of fibroblasts and extracellular matrix components, as well as the expression of characteristic genes and secretion of chemokines, cytokines, and growth factors. AACT grafts were tested in the rabbits from which the fibroblasts were obtained, whereas acellular collagen type I scaffolds (CS) were evaluated in a separate group of rabbits. In both cases, contralateral wounds closed by secondary intention were used as controls. In a separate experiment, AACT-grafted wounds were directly compared with contralateral CS-grafted wounds in the same animals. Wound contraction and histological parameters were examined to evaluate closure differences between the treatments in the three animal experiments performed. Contraction of wounds grafted with AACT and CS was significantly lower than in their controls (p oral mucosa.

  1. 以猪角膜脱细胞基质为载体培养人脐静脉内皮细胞构建实验性角膜后板层%Decellularized porcine corneal posterior lamellae as carrier matrix for cultivating human umbilical vein endothelial cells in vitro

    Institute of Scientific and Technical Information of China (English)

    祁冰; 侯光辉; 李柳; 季青山; 吴静; 周清

    2013-01-01

    AIM:To investigate the feasibility of corneal posterior lamellar reconstruction with human umbilical vein endothelial cells (HUVECs) and porcine cornea acellular matrix in vitro,and to observe the physiological function of the transplantation in vivo.METHODS:HUVECs were isolated,cultured,and labeled with fluorescent dye CM-DiI.Porcine corneas were treated with 100% glycerinum,cut to a thinner structure step by step,and dried on the super-clean bench.Transmission electron microscope were used to observe the histological changes of the porcine cornes acellular matrix.Labeled HUVECs were seeded onto the porcine cornea acellular matrix,and examined by scanning electron microscopy.When the HUVECs and Descemet's membrane fusion formed a monolayer,the corneal transplantation in rabbits was performed.Twenty-four New Zealand white rabbits were randomly divided into experimental group and control group (n =12 each),and their left eyes served as recipients.RESULTS:Cultured HUVECs exhibited polygonal shape.More than 90% HUVECs were labeled with CM-DiI and the cell membrane was positive with red fluorescence,which was detectable at least up to 3 generations.The histological examination indicated that porcine cornea cells were clearly extracted,and the collagen fibers were well arranged.A continuous monolayer of HUVECs on the porcine cornea acellular matrix was observedunder scanning electron microscopy.The reconstructed corneal posterior lamellae were similar to the noral cornea.The observation of transplantation showed that the cornea in experimental group was substantially transparent.However,that in control group was oedematous and adiaphanous.CONCLUSION:Corneal posterior lamellae can be reconstructedin vitro by cultivating HUVECs on porcine cornea acellular matrix.After xenogeneic transplantation,the graft survivesin vivo and expresses normal corneal endothelial cell biological functions.Deep lamellar corneal endothelial transplantation is an effective keratoplasty.%

  2. Inhibition of matrix metalloproteinase activity in human dentin via novel antibacterial monomer

    Science.gov (United States)

    Li, Fang; Majd, Hessam; Weir, Michael D.; Arola, Dwayne D.; Xu, Hockin H.K.

    2015-01-01

    Objectives Dentin-composite bond failure is caused by factors including hybrid layer degradation, which in turn can be caused by hydrolysis and enzymatic degradation of the exposed collagen in the dentin. The objectives of this study were to investigate a new antibacterial monomer (dimethylaminododecyl methacrylate, DMADDM) as an inhibitor for matrix metalloproteinases (MMPs), and to determine the effects of DMADDM on both soluble recombinant human MMPs (rhMMPs) and dentin matrix-bound endogenous MMPs. Methods Inhibitory effects of DMADDM at six mass% (0.1% to 10%) on soluble rhMMP-8 and rhMMP-9 were measured using a colorimetic assay. Matrix-bound endogenous MMP activity was evaluated in demineralized human dentin. Dentin beams were divided into four groups (n = 10) and incubated in calcium- and zinc-containing media (control medium); or control medium + 0.2% chlorhexidine (CHX); 5% 12-methacryloyloxydodecylpyridinium bromide (MDPB); or 5% DMADDM. Dissolution of dentin collagen peptides was evaluated by mechanical testing in three-point flexure, loss of dentin mass, and a hydroxyproline assay. Results Use of 0.1% to 10% DMADDM exhibited a strong concentration-dependent anti-MMP effect, reaching 90% of inhibition on rhMMP-8 and rhMMP-9 at 5% DMADDM concentration. Dentin beams in medium with 5% DMADDM showed 34% decrease in elastic modulus (vs. 73% decrease for control), 3% loss of dry dentin mass (vs. 28% loss for control), and significantly less solubilized hydroxyproline when compared with control (p dentin MMPs. These results, together with previous studies showing that adhesives containing DMADDM inhibited biofilms without compromising dentin bond strength, suggest that DMADDM is promising for use in adhesives to prevent collagen degradation in hybrid layer and protect the resin-dentin bond. PMID:25595564

  3. Human Kunitz-type protease inhibitor engineered for enhanced matrix retention extends longevity of fibrin biomaterials.

    Science.gov (United States)

    Briquez, Priscilla S; Lorentz, Kristen M; Larsson, Hans M; Frey, Peter; Hubbell, Jeffrey A

    2017-08-01

    Aprotinin is a broad-spectrum serine protease inhibitor used in the clinic as an anti-fibrinolytic agent in fibrin-based tissue sealants. However, upon re-exposure, some patients suffer from hypersensitivity immune reactions likely related to the bovine origin of aprotinin. Here, we aimed to develop a human-derived substitute to aprotinin. Based on sequence homology analyses, we identified the Kunitz-type protease inhibitor (KPI) domain of human amyloid-β A4 precursor protein as being a potential candidate. While KPI has a lower intrinsic anti-fibrinolytic activity than aprotinin, we reasoned that its efficacy is additionally limited by its fast release from fibrin material, just as aprotinin's is. Thus, we engineered KPI variants for controlled retention in fibrin biomaterials, using either covalent binding through incorporation of a substrate for the coagulation transglutaminase Factor XIIIa or through engineering of extracellular matrix protein super-affinity domains for sequestration into fibrin. We showed that both engineered KPI variants significantly slowed plasmin-mediated fibrinolysis in vitro, outperforming aprotinin. In vivo, our best engineered KPI variant (incorporating the transglutaminase substrate) extended fibrin matrix longevity by 50%, at a dose at which aprotinin did not show efficacy, thus qualifying it as a competitive substitute of aprotinin in fibrin sealants. Copyright © 2017 The Author(s). Published by Elsevier Ltd.. All rights reserved.

  4. Development and characterization of decellularized human nasoseptal cartilage matrix for use in tissue engineering.

    Science.gov (United States)

    Graham, M Elise; Gratzer, Paul F; Bezuhly, Michael; Hong, Paul

    2016-10-01

    Reconstruction of cartilage defects in the head and neck can require harvesting of autologous cartilage grafts, which can be associated with donor site morbidity. To overcome this limitation, tissue-engineering approaches may be used to generate cartilage grafts. The objective of this study was to decellularize and characterize human nasoseptal cartilage with the aim of generating a biological scaffold for cartilage tissue engineering. Laboratory study using nasoseptal cartilage. Remnant human nasoseptal cartilage specimens were collected and subjected to a novel decellularization treatment. The decellularization process involved several cycles of enzymatic detergent treatments. For characterization, decellularized and fresh (control) specimens underwent histological, biochemical, and mechanical analyses. Scanning electron microscopy and biocompatibility assay were also performed. The decellularization process had minimal effect on glycosaminoglycan content of the cartilage extracellular matrix. Deoxyribonucleic acid (DNA) analysis revealed the near-complete removal of genomic DNA from decellularized tissues. The effectiveness of the decellularization process was also confirmed on histological and scanning electron microscopic analyses. Mechanical testing results showed that the structural integrity of the decellularized tissue was maintained, and biocompatibility was confirmed. Overall, the current decellularization treatment resulted in significant reduction of genetic/cellular material with preservation of the underlying extracellular matrix structure. This decellularized material may serve as a potential scaffold for cartilage tissue engineering. N/A. Laryngoscope, 126:2226-2231, 2016. © 2016 The American Laryngological, Rhinological and Otological Society, Inc.

  5. The enamel matrix derivative (Emdogain) enhances human tongue carcinoma cells gelatinase production, migration and metastasis formation.

    Science.gov (United States)

    Laaksonen, Matti; Suojanen, Juho; Nurmenniemi, Sini; Läärä, Esa; Sorsa, Timo; Salo, Tuula

    2008-08-01

    Enamel matrix derivative Emdogain (EMD) is widely used in periodontal treatment to regenerate lost connective tissue and to improve the attachment of the teeth. Gelatinases (MMP-2 and -9) have an essential role in the promotion and progression of oral cancer growth and metastasis formation. We studied the effects of EMD on human tongue squamous cell carcinoma (HSC-3) cells in vitro and in vivo. In vitro, EMD (100 microg/ml and 200 microg/ml) remarkably induced the MMP-2 and -9 production from HSC-3 cells analysed by zymography and enzyme-linked immunosorbent assay. EMD also slightly induced the MMP-2 and -9 production from benign human mucosal keratinocytes (HMK). Furthermore, EMD clearly induced the transmigration of HSC-3 cells but had no effect on the HMK migration in transwell assays. The in vitro wound closure of HSC-3 cells was notably accelerated by EMD, whereas it had only minor effect on the wound closure of HMKs. The migration of both cell lines was inhibited by a selective cyclic anti-gelatinolytic peptide CTT-2. EMD had no effect on HSC-3 cell proliferation or apoptosis and only a limited effect on cell attachment to various extracellular matrix components. The in vivo mice experiment revealed that EMD substantially induced HSC-3 xenograft metastasis formation. Our results suggest that the use of EMD for patients with oral mucosal carcinomas or premalignant lesions should be carefully considered, possibly avoided.

  6. Effect of Supracervical Apposition and Spontaneous Labour on Apoptosis and Matrix Metalloproteinases in Human Fetal Membranes

    Directory of Open Access Journals (Sweden)

    Mahalia Chai

    2013-01-01

    Full Text Available Background. Apoptosis and matrix metalloproteinase (MMP-9 are capable of hydrolysing components of the extracellular matrix and weakening the fetal membranes which leads to eventual rupture, a key process of human parturition. The aim of this study was to determine the effect of supracervical apposition and spontaneous labour on apoptosis and MMP-9 in human fetal membranes at term. Methods. Fetal membranes were obtained from term non-labouring supracervical site (SCS and compared to (i a paired distal site (DS or (ii site of rupture (SOR after spontaneous labour onset. Results. The expression of the proapoptotic markers Bax, Smac, Fas, FasL, caspase-3, and PARP, was significantly higher in the non-labouring SCS chorion compared to paired DS. Bax, Smac, FasL, caspase-3, and PARP staining was higher in the non-labouring SCS fetal membranes than that in the post-labour SOR. MMP-9 expression and activity were higher in the post-labour SOR fetal membranes compared to non-labouring SCS fetal membranes. Conclusion. Components of the apoptotic signalling pathways and MMP-9 may play a role in rupture and labour. Non-labouring SCS fetal membranes display altered morphology and altered apoptotic biochemical characteristics in preparation for labour, while the laboured SOR displays unique MMP characteristics.

  7. Risk of Febrile Seizures and Epilepsy After Vaccination With Diphtheria, Tetanus, Acellular Pertussis, Inactivated Poliovirus, and Haemophilus Influenzae Type b

    DEFF Research Database (Denmark)

    Sun, Yuelian; Christensen, Jakob Christensen; Hviid, Anders

    2012-01-01

    Context Vaccination with whole-cell pertussis vaccine carries an increased risk of febrile seizures, but whether this risk applies to the acellular pertussis vaccine is not known. In Denmark, acellular pertussis vaccine has been included in the combined diphtheria-tetanus toxoids-acellular pertus......Context Vaccination with whole-cell pertussis vaccine carries an increased risk of febrile seizures, but whether this risk applies to the acellular pertussis vaccine is not known. In Denmark, acellular pertussis vaccine has been included in the combined diphtheria-tetanus toxoids...

  8. Scanning Electron Microscopic Examination of the Extracellular Matrix in the Decellularized Mouse and Human Cochlea.

    Science.gov (United States)

    Santi, Peter A; Aldaya, Robair; Brown, Alec; Johnson, Shane; Stromback, Tyler; Cureoglu, Sebahattin; Rask-Andersen, Helge

    2016-06-01

    Decellularized tissues have been used to investigate the extracellular matrix (ECM) in a number of different tissues and species. Santi and Johnson JARO 14:3-15 (2013) first described the decellularized inner ear in the mouse, rat, and human using scanning thin-sheet laser imaging microscopy (sTSLIM). The purpose of the present investigation is to examine decellularized cochleas in the mouse and human at higher resolution using scanning electron microscopy (SEM). Fresh cochleas were harvested and decellularized using detergent extraction methods. Following decellularization, the ECM of the bone, basilar membrane, spiral limbus, and ligament remained, and all of the cells were removed from the cochlea. A number of similarities and differences in the ECM of the mouse and human were observed. A novel, spirally directed structure was present on the basilar membrane and is located at the border between Hensen and Boettcher cells. These septa-like structures formed a single row in the mouse and multiple rows in the human. The basal lamina of the stria vascularis capillaries was present and appeared thicker in the human compared with the mouse. In the mouse, numerous openings beneath the spiral prominence that previously housed the root processes of the external sulcus cells were observed but in the human there was only a single row of openings. These and other anatomical differences in the ECM between the mouse and human may reflect functional differences and/or be due to aging; however, decellularized cochleas provide a new way to examine the cochlear ECM and reveal new observations.

  9. Suppression of α Smooth Muscle Actin Accumulation by Bovine Fetal Dermal Collagen Matrix in Full Thickness Skin Wounds.

    Science.gov (United States)

    Lineaweaver, William; Bush, Katie; James, Kenneth

    2015-06-01

    The suppression of elements associated with wound contracture and unfavorable scarring is a potentially important strategy in clinical wound management. In this study, the presence of α smooth muscle actin (αSMA), a protein involved in wound contraction, was analyzed in a series of wounds in which bovine fetal collagen (BFC) acellular dermal matrix (PriMatrix) was used in staged split thickness skin graft procedures. The results obtained through histological and quantitative image analyses of incidental biopsies from these wounds demonstrated a suppression of αSMA in the wound regions occupied by assimilated BFC relative to increased levels of αSMA found in other areas of the wound. The αSMA levels found in assimilated BFC were similar to αSMA levels in uninjured human dermis. These findings suggest a mechanism by which application of BFC could decrease contraction of full thickness skin wounds.

  10. Development of a porcine renal extracellular matrix scaffold as a platform for kidney regeneration.

    Science.gov (United States)

    Choi, Seock Hwan; Chun, So Young; Chae, Seon Yeong; Kim, Jin Rae; Oh, Se Heang; Chung, Sung Kwang; Lee, Jin Ho; Song, Phil Hyun; Choi, Gyu-Seog; Kim, Tae-Hwan; Kwon, Tae Gyun

    2015-04-01

    Acellular scaffolds, possessing an intact three-dimensional extracellular matrix (ECM) architecture and biochemical components, are promising for regeneration of complex organs, such as the kidney. We have successfully developed a porcine renal acellular scaffold and analyzed its physical/biochemical characteristics, biocompatibility, and kidney reconstructive potential. Segmented porcine kidney cortexes were treated with either 1% (v/v) Triton X-100 (Triton) or sodium dodecyl sulfate (SDS). Scanning electron microscopy showed both treatments preserved native tissue architecture, including porosity and composition. Swelling behavior was higher in the Triton-treated compared with the SDS-treated scaffold. Maximum compressive strength was lower in the Triton-treated compared with the SDS-treated scaffold. Attenuated total reflective-infrared spectroscopy showed the presence of amide II (-NH) in both scaffolds. Furthermore, richer ECM protein and growth factor contents were observed in the Triton-treated compared with SDS-treated scaffold. Primary human kidney cell adherence, viability, and proliferation were enhanced on the Triton-treated scaffold compared with SDS-treated scaffold. Following murine in vivo implantation, tumorigenecity was absent for both scaffolds after 8 weeks and in the Triton-treated scaffold only, glomeruli-like structure formation and neovascularity were observed. We identified 1% Triton X-100 as a more suitable decellularizing agent for porcine renal ECM scaffolds prior to kidney regeneration.

  11. Substratum Stiffness and Latrunculin B Regulate Matrix Gene and Protein Expression in Human Trabecular Meshwork Cells

    Science.gov (United States)

    Thomasy, Sara M.; Wood, Joshua A.; Kass, Philip H.; Murphy, Christopher J.

    2012-01-01

    Purpose. To determine the impact of substratum stiffness and latrunculin-B (Lat-B), on the expression of several matrix proteins that are associated with glaucoma. Methods. Human trabecular meshwork (HTM) cells were cultured on hydrogels possessing stiffness values mimicking those found in normal (5 kPa) and glaucomatous meshworks (75 kPa), or tissue culture polystyrene (TCP; >1 GPa). Cells were treated with 2.0 μM Lat-B in dimethyl sulfoxide (DMSO) or DMSO alone. RT-PCR was used to determine the impact of substratum stiffness and/or Lat-B treatment on the expression of secreted protein, acidic, cysteine rich (SPARC), myocilin, angiopoietin-like factor (ANGPTL)-7, and transglutaminase (TGM)-2. Immunofluorescence was used to assess changes in protein expression. Results. SPARC and myocilin mRNA expression were dramatically increased on the 75 kPa hydrogels and decreased on the 5 kPa hydrogels in comparison to TCP. In contrast, ANGPTL-7 mRNA and TGM-2 mRNA was decreased on the 75 kPa and 5 kPa hydrogels, respectively, in comparison with TCP. Treatment with Lat-B dramatically downregulated both SPARC and myocilin on 75 kPa hydrogels. In contrast, cells grown on TCP produced greater or similar amounts of SPARC and myocilin mRNA after Lat-B treatment. SPARC and myocilin protein expression paralleled changes in mRNA expression. Conclusions. Substratum stiffness impacts HTM matrix gene and protein expression and modulates the impact of Lat-B treatment on the expression of these matrix proteins. Integrating the use of biologically relevant substratum stiffness in the conduction of in vitro experiments gives important insights into HTM cell response to drugs that may more accurately predict responses observed in vivo. PMID:22247475

  12. The upregulation of matrix metalloproteinase -2 and -9 genes caused by resistin in human chondrocytes

    Directory of Open Access Journals (Sweden)

    Kürşat Oğuz Yaykaşlı

    2014-02-01

    Full Text Available  OBJECTIVES: The articular cartilage allows movement by absorbing mechanical loading within a physiological range. However, the accumulation of excessive adipose tissue has catabolic effect on extracellular matrix (ECM components in some diseases such as rheumatoid arthritis (RA and osteoarthritis (OA. Resistin, inflammatory adipokines is secreted by adipose tissue, and the elevated serum level was reported in obese subjects and patients with RA and OA. Gelatinases (MMP-2 and MMP-9, a subfamily of matrix metalloproteinases are responsible for destruction of collagen and matrix components. In this study, the effect of resistin on gelatinases genes expression was investigated. METHODS: Human chondrocytes was stimulated by resistin at 100 and 250 ng/ml doses for 3h, 6h, 12h, 24h and 48h. 2 µg RNA was subject to reverse transcription after RNA extraction. Gelatinases expressions were analyzed by quantitative Real-Time Polymerase Chain Reaction method. RESULTS: The expression levels of gelatinases were increased at both 100 and 250 ng/mL and peaked at 250 ng/ml dose for 48 hours. CONCLUSIONS: The clarification of etiology for irreversible destruction of ECM has a vital importance to develop new treatment strategies for RA and OA. In conclusion, increased levels of gelatinases expression caused by resistin were founded. The upregulation of gelatinases caused by resistin is might be a new target for obesity associated patients with RA and OA. However, the molecular pathways of this induction should be investigated in other studies. 

  13. Effects of N-acetylcysteine on matrix metalloproteinase-9 secretion and cell migration of human corneal epithelial cells

    OpenAIRE

    Ramaesh, T; Ramaesh, K; Riley, S C; West, J.D.; Dhillon, B

    2012-01-01

    Matrix metalloproteinase-9 (MMP-9) secreted by corneal epithelial cells has a role in the remodelling of extracellular matrix and migration of epithelial cells. Elevated levels of MMP-9 activity in the ocular surface may be involved in the pathogenesis of corneal diseases. N-acetylcysteine (NAC) has been used to treat corneal diseases, including recurrent epithelial erosions. In this study, its effects on the MMP-9 secretion and human corneal epithelial (HCE) cell migration were evaluated in ...

  14. Structural characterization of membrane-bound human immunodeficiency virus-1 Gag matrix with neutron reflectometry

    Science.gov (United States)

    Eells, Rebecca; Barros, Marilia; Scott, Kerry M.; Karageorgos, Ioannis; Heinrich, Frank; Lösche, Mathias

    2017-01-01

    The structural characterization of peripheral membrane proteins represents a tremendous challenge in structural biology due to their transient interaction with the membrane and the potential multitude of protein conformations during this interaction. Neutron reflectometry is uniquely suited to address this problem because of its ability to structurally characterize biological model systems nondestructively and under biomimetic conditions that retain full protein functionality. Being sensitive to only the membrane-bound fraction of a water-soluble peripheral protein, neutron reflectometry obtains a low-resolution average structure of the protein-membrane complex that is further refined using integrative modeling strategies. Here, the authors review the current technological state of biological neutron reflectometry exemplified by a detailed report on the structure determination of the myristoylated human immunodeficiency virus-1 (HIV-1) Gag matrix associated with phosphoserine-containing model membranes. The authors found that the HIV-1 Gag matrix is able to adopt different configurations at the membrane in a pH-dependent manner and that the myristate group orients the protein in a way that is conducive to PIP2-binding. PMID:28511544

  15. Volumetric characterization of human patellar cartilage matrix on phase contrast x-ray computed tomography

    Science.gov (United States)

    Abidin, Anas Z.; Nagarajan, Mahesh B.; Checefsky, Walter A.; Coan, Paola; Diemoz, Paul C.; Hobbs, Susan K.; Huber, Markus B.; Wismüller, Axel

    2015-03-01

    Phase contrast X-ray computed tomography (PCI-CT) has recently emerged as a novel imaging technique that allows visualization of cartilage soft tissue, subsequent examination of chondrocyte patterns, and their correlation to osteoarthritis. Previous studies have shown that 2D texture features are effective at distinguishing between healthy and osteoarthritic regions of interest annotated in the radial zone of cartilage matrix on PCI-CT images. In this study, we further extend the texture analysis to 3D and investigate the ability of volumetric texture features at characterizing chondrocyte patterns in the cartilage matrix for purposes of classification. Here, we extracted volumetric texture features derived from Minkowski Functionals and gray-level co-occurrence matrices (GLCM) from 496 volumes of interest (VOI) annotated on PCI-CT images of human patellar cartilage specimens. The extracted features were then used in a machine-learning task involving support vector regression to classify ROIs as healthy or osteoarthritic. Classification performance was evaluated using the area under the receiver operating characteristic (ROC) curve (AUC). The best classification performance was observed with GLCM features correlation (AUC = 0.83 +/- 0.06) and homogeneity (AUC = 0.82 +/- 0.07), which significantly outperformed all Minkowski Functionals (p GLCM-derived statistical features can distinguish between healthy and osteoarthritic tissue with high accuracy.

  16. Photobiomodulation on human annulus fibrosus cells during the intervertebral disk degeneration: extracellular matrix-modifying enzymes.

    Science.gov (United States)

    Hwang, Min Ho; Kim, Kyoung Soo; Yoo, Chang Min; Shin, Jae Hee; Nam, Hyo Geun; Jeong, Jin Su; Kim, Joo Han; Lee, Kwang Ho; Choi, Hyuk

    2016-05-01

    Destruction of extracellular matrix (ECM) leads to degeneration of the intervertebral disk (IVD), which is a major contributor to many spine disorders. IVD degeneration is induced by pro-inflammatory cytokines, such as tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β), which are secreted by immune cells, including macrophages and neutrophils. The cytokines modulate ECM-modifying enzymes such as matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in human annulus fibrosus (AF) cells. The resulting imbalance in catabolic and anabolic enzymes can cause generalized back, neck, and low back pain (LBP). Photobiomodulation (PBM) is known to regulate inflammatory responses and wound healing. The aim of this study was to mimic the degenerative IVD microenvironment, and to investigate the effect of a variety of PBM conditions (wavelength: 635, 525, and 470 nm; energy density: 16, 32, and 64 J/cm(2)) on the production of ECM-modifying-enzymes by AF cells under degenerative conditions induced by macrophage-conditioned medium (MCM), which contains pro-inflammatory cytokines such as TNF-α and IL-β secreted by macrophage during the development of intervertebral disk inflammation. We showed that the MCM-stimulated AF cells express imbalanced ratios of TIMPs (TIMP-1 and TIMP-2) and MMPs (MMP-1 and MMP-3). PBM selectively modulated the production of ECM-modifying enzymes in AF cells. These results suggest that PBM can be a therapeutic tool for degenerative IVD disorders.

  17. Food Matrix Effects of Polyphenol Bioaccessibility from Almond Skin during Simulated Human Digestion

    Directory of Open Access Journals (Sweden)

    Giuseppina Mandalari

    2016-09-01

    Full Text Available The goal of the present study was to quantify the rate and extent of polyphenols released in the gastrointestinal tract (GIT from natural (NS and blanched (BS almond skins. A dynamic gastric model of digestion which provides a realistic simulation of the human stomach was used. In order to establish the effect of a food matrix on polyphenols bioaccessibility, NS and BS were either digested in water (WT or incorporated into home-made biscuits (HB, crisp-bread (CB and full-fat milk (FM. Phenolic acids were the most bioaccessible class (68.5% release from NS and 64.7% from BS. WT increased the release of flavan-3-ols (p < 0.05 and flavonols (p < 0.05 from NS after gastric plus duodenal digestion, whereas CB and HB were better vehicles for BS. FM lowered the % recovery of polyphenols, the free total phenols and the antioxidant status in the digestion medium, indicating that phenolic compounds could bind protein present in the food matrix. The release of bioactives from almond skins could explain the beneficial effects associated with almond consumption.

  18. Grooved surface topography alters matrix-metalloproteinase production by human fibroblasts

    Energy Technology Data Exchange (ETDEWEB)

    Brydone, Alistair S; Dominic Meek, R M [Department of Orthopaedics, Southern General Hospital, 1345 Govan Road, Glasgow G51 4TF (United Kingdom); Dalby, Matthew J; Berry, Catherine C; McNamara, Laura E, E-mail: alibrydone@gmail.com [Centre for Cell Engineering, Joseph Black Building, Institute of Molecular, Cell and Systems Biology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ (United Kingdom)

    2011-06-15

    Extracellular matrix (ECM) remodelling is an essential physiological process in which matrix-metalloproteinases (MMPs) have a key role. Manipulating the manner in which cells produce MMPs and ECMs may enable the creation of a desired tissue type, i.e. effect repair, or the prevention of tissue invasion (e.g. metastasis). The aim of this project was to determine if culturing fibroblasts on grooved topography altered collagen deposition or MMP production. Human fibroblasts were seeded on planar or grooved polycaprolactone substrates (grooves were 12.5 {mu}m wide with varying depths of 240 nm, 540 nm or 2300 nm). Cell behaviour and collagen production were studied using fluorescence microscopy and the spent culture medium was assessed using gel zymography to detect MMPs. Total collagen deposition was high on the 240 nm deep grooves, but decreased as the groove depth increased, i.e. as cell contact guidance decreased. There was an increase in gelatinase on the 2300 nm deep grooved topography and there was a difference in the temporal expression of MMP-3 observed on the planar surface compared to the 540 nm and 2300 nm topographies. These results show that topography can alter collagen and MMP production. A fuller understanding of these processes may permit the design of surfaces tailored to tissue regeneration e.g. tendon repair.

  19. Host-Parasite Interaction: Parasite-Derived and -Induced Proteases That Degrade Human Extracellular Matrix

    Directory of Open Access Journals (Sweden)

    Carolina Piña-Vázquez

    2012-01-01

    Full Text Available Parasitic protozoa are among the most important pathogens worldwide. Diseases such as malaria, leishmaniasis, amoebiasis, giardiasis, trichomoniasis, and trypanosomiasis affect millions of people. Humans are constantly threatened by infections caused by these pathogens. Parasites engage a plethora of surface and secreted molecules to attach to and enter mammalian cells. The secretion of lytic enzymes by parasites into host organs mediates critical interactions because of the invasion and destruction of interstitial tissues, enabling parasite migration to other sites within the hosts. Extracellular matrix is a complex, cross-linked structure that holds cells together in an organized assembly and that forms the basement membrane lining (basal lamina. The extracellular matrix represents a major barrier to parasites. Therefore, the evolution of mechanisms for connective-tissue degradation may be of great importance for parasite survival. Recent advances have been achieved in our understanding of the biochemistry and molecular biology of proteases from parasitic protozoa. The focus of this paper is to discuss the role of protozoan parasitic proteases in the degradation of host ECM proteins and the participation of these molecules as virulence factors. We divide the paper into two sections, extracellular and intracellular protozoa.

  20. Enamel Matrix Derivative Promote Primary Human Pulp Cell Differentiation and Mineralization

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    Elisabeth Aurstad Riksen

    2014-05-01

    Full Text Available Enamel matrix derivative (EMD has been found to induce reactive dentin formation; however the molecular mechanisms involved are unclear. The effect of EMD (5–50 μg/mL on primary human pulp cells were compared to untreated cells and cells incubated with 10−8 M dexamethasone (DEX for 1, 2, 3, 7, and 14 days in culture. Expression analysis using Affymetrix microchips demonstrated that 10 μg/mL EMD regulated several hundred genes and stimulated the gene expression of proteins involved in mesenchymal proliferation and differentiation. Both EMD and DEX enhanced the expression of amelogenin (amel, and the dentinogenic markers dentin sialophosphoprotein (DSSP and dentin matrix acidic phosphoprotein 1 (DMP1, as well as the osteogenic markers osteocalcin (OC, BGLAP and collagen type 1 (COL1A1. Whereas, only EMD had effect on alkaline phosphatase (ALP mRNA expression, the stimulatory effect were verified by enhanced secretion of OC and COL1A from EMD treated cells, and increased ALP activity in cell culture medium after EMD treatment. Increased levels of interleukin-6 (IL-6, interleukin-8 (IL-8, and monocyte chemoattractant proteins (MCP-1 in the cell culture medium were also found. Consequently, the suggested effect of EMD is to promote differentiation of pulp cells and increases the potential for pulpal mineralization to favor reactive dentine formation.

  1. 珊瑚羟基磷灰石与异体脱细胞真皮基质联合修复牙根尖周组织缺损%Acellular dermal matrix allograft combined with coralline hydroxyapatite repair periapical tissue defects

    Institute of Scientific and Technical Information of China (English)

    徐隽; 王进涛; 李刚; 史芳川; 钟良军

    2014-01-01

      结果与结论:修复1个月后,实验组患者异体脱细胞真皮基质全部存活,因修整瘘管口周围炎性的肉芽组织导致的牙龈组织缺损已经愈合。在修复12个月后,实验组患者的修复有效率明显高于对照组(P <0.05)。实验组患者修复6个月后骨缺损区阴影基本消失,珊瑚羟基磷灰石颗粒间的透射影减小,出现有一定致密度的影像,提示有新骨长入;12个月后珊瑚羟基磷灰石颗粒密度已接近正常的骨组织密度,与正常骨组织之间有密度移行改变,逐渐与牙槽骨形成骨融合。异体脱细胞基质与珊瑚羟基磷灰石的生物相容性良好。提示异体脱细胞真皮基质与珊瑚羟基磷灰石联合修复根尖周组织缺损具有良好的临床疗效。%Chronic periapical periodontitis often causes periapical tissue defects and ultimately leads to the loss of teeth if the inflammation is not promptly cleared to terminate bone resorption and destruction of gingival tissue. Acelular dermal matrix alograft and coraline hydroxyapatite are the common materials to repair periodontal injury. To evaluate clinical efficacy of acelular dermal matrix alograft combined with coraline hydroxyapatite in repairing periapical tissue defects. A total of 76 patients of chronic apical periodontitis were randomly divided into two groups, with 38 cases in each group. In the experimental group, periapical tissue defects were treated with acelular dermal matrix alograft and coraline hydroxyapatite. In the control group, tissue defects were not treated. Al the involved patients underwent apicectomy and retrograde filing. Clinical parameters and radiographic film were recorded at 1 week, 6 months and 3 years folow-up visits to evaluate the repairing effects. After 1 month of treatment, al acelular dermal matrix alografts survived, and the defect of gingival tissues that caused by repairing fistula had been healed. After 3 years, the repairing

  2. Human cardiac extracellular matrix supports myocardial lineage commitment of pluripotent stem cells

    DEFF Research Database (Denmark)

    Oberwallner, Barbara; Brodarac, Andreja; Anić, Petra;

    2015-01-01

    lysis buffer, sodium dodecyl sulphate (SDS) and foetal bovine serum (FBS). Murine embryonic stem cells (ESCs), induced pluripotent stem cells (iPSCs) and mesenchymal stromal cells (MSCs) were seeded and grown in standard culture, on cECM or on non-specific ECM preparations (Matrigel® or Geltrex®). Cell......OBJECTIVES: Cross-talk between organ-specific extracellular matrix (ECM) and stem cells is often assumed but has not been directly demonstrated. We developed a protocol for the preparation of human cardiac ECM (cECM) and studied whether cECM has effects on pluripotent stem cell differentiation...... that may be useful for future cardiac regeneration strategies in patients with end-stage heart failure. METHODS: Of note, 0.3 mm-thick cECM slices were prepared from samples of myocardium from patients with end-stage non-ischaemic dilated cardiomyopathy, using a three-step protocol involving hypotonic...

  3. Changes of Nuclear Matrix Proteins Following the Differentiation of Human Osteosarcoma MG-63 Cells

    Institute of Scientific and Technical Information of China (English)

    Chun-Hong Zhao; Qi-Fu Li; Yan Zhao; Jing-Wen Niu; Zhi-Xing Li; Jin-An Chen

    2006-01-01

    Human osteosarcoma MG-63 cells were induced into differentiation by 5 mmol/L hexamethylene bisacetamide (HMBA). Their nuclear matrix proteins (NMPs) were selectively extracted and subjected to two-dimensional gel electrophoresis analysis.The results of protein patterns were analyzed by Melanie software. The spots of differentially expressed NMPs were excised and subjected to in situ digestion with trypsin. The maps of peptide mass fingerprinting were obtained by MALDI-TOFMS analysis, and were submitted for NCBI database searches by Mascot tool.There were twelve spots changed remarkably during the differentiation induced by HMBA, nine of which were identified. The roles of the regulated proteins during the MG-63 differentiation were analyzed. This study suggests that the induced differentiation of cancer cells is accompanied by the changes of NMPs, and confirms the presence of some specific NMPs related to the cancer cell proliferation and differentiation. The changed NMPs are potential markers for cancer diagnosis or targets for cancer therapy.

  4. REACTOGENICITY OF ACELLULAR PERTUSSIS VACCINE AND THE POSSIBILITY OF ITS USE IN ELDER CHILDREN

    Directory of Open Access Journals (Sweden)

    M.G. Galitskaya

    2008-01-01

    Full Text Available As is know, in the past few years, the incidence of pertussis has increased again. The article reveals the reasons of this phenomenon and the possible solutions for this problem. Besides, comparative analysis of the whole cell vaccine used in this country as within the framework of the national immunizations schedule and modern acellular vaccines is made. Results of multicenter research, convincingly proving the safety and efficiency of acellular pertussis vaccine, are presented.Key words: pertussis, prophylaxis, whole cell vaccine, acellular vaccine, efficiency, children.

  5. Expression of matrix metrallproteinase-2 in human tears fluid after LASIK

    Directory of Open Access Journals (Sweden)

    Ai-Wei Chen

    2014-12-01

    Full Text Available AIM: To monitor long-term changes of matrix metalloproteinase-2(MMP-2in human tears fluid after laser in situ keratomileusis(LASIK. METHODS: Thirty-two myopia cases(64 eyesunderwent uneventful LASIK were enrolled in the study. Tear fluid were collected and MMP-2 expression was analyzed by Western-bolt assay preoperatively and postoperatively on 15d, at 1, 3mo, and 1a. RESULTS: LASIK increased the concentration of MMP-2 in human tear fluid. At 15d postoperatively, the magnitude of MMP-2 was 1.4 times that of preoperative, thereafter subsided, but didn't return to preoperative level by 3mo(PP>0.05. CONCLUSION: MMP-2 is significantly expressed in human tear fluid after LASIK, then subsided with time, but didn't return to preoperative level by 3mo and almost recovered up to 1a, indicating wound healing of LASIK would continue up at least 3mo after surgery and almost recovered 1a postoperatively.

  6. Mineralized polycaprolactone nanofibrous matrix for odontogenesis of human dental pulp cells.

    Science.gov (United States)

    Kim, Jong-Jin; Bae, Won-Jung; Kim, Joung-Mok; Kim, Jung-Ju; Lee, Eun-Jung; Kim, Hae-Won; Kim, Eun-Cheol

    2014-03-01

    The aim of the present study was to fabricate mineralized polycaprolactone nanofibrous scaffold and investigate its ability to elicit odontogenic differentiation of human dental pulp cells, compared to the pure polycaprolactone scaffold. Polycaprolactone nanofibrous scaffold was produced by electrospinning, and the surface was mineralized with apatite. Cellular behaviors on the mineralized polycaprolactone scaffold were assessed in terms of cell adhesion, growth, and odontoblastic differentiation. To evaluate the signal transduction of human dental pulp cells, mRNA expression was analyzed and Western blotting was performed. Mineralized polycaprolactone showed improved cell proliferation, mineralized nodule formation, and expression of odontoblastic marker genes including alkaline phosphatase, osteopontin, osteocalcin, dentin sialophosphoprotein (DSPP), and dentin matrix protein-1, as compared with pure polycaprolactone. Although the cell adhesion on the mineralized polycaprolactone was similar to that of the polycaprolactone, the expression level of proteins including collagen type I and the key adhesion receptor (integrin components α1, α2, and β1) was upregulated in mineralized polycaprolactone compared to polycaprolactone. Especially, cells seeded onto mineralized polycaprolactone scaffolds showed significantly increased levels of phosphorylated focal adhesion kinase, a marker of integrin activation, and downstream pathways, such as phosphor (p)-Akt, p-extracellular signal regulated kinase, p-c Jun N-terminal kinase, nuclear factor-kappa B, c-fos, and c-jun, compared with pure polycaprolactone. The mineralized polycaprolactone scaffold is attractive for dentin tissue engineering by promoting growth and odontogenic differentiation of human dental pulp cells through the integrin-mediated signaling pathway.

  7. Human Dupuytren's Ex Vivo Culture for the Study of Myofibroblasts and Extracellular Matrix Interactions.

    Science.gov (United States)

    Karkampouna, Sofia; Kloen, Peter; Obdeijn, Miryam C; Riester, Scott M; van Wijnen, Andre J; Kruithof-de Julio, Marianna

    2015-01-01

    Organ fibrosis or "scarring" is known to account for a high death toll due to the extensive amount of disorders and organs affected (from cirrhosis to cardiovascular diseases). There is no effective treatment and the in vitro tools available do not mimic the in vivo situation rendering the progress of the out of control wound healing process still enigmatic. To date, 2D and 3D cultures of fibroblasts derived from DD patients are the main experimental models available. Primary cell cultures have many limitations; the fibroblasts derived from DD are altered by the culture conditions, lack cellular context and interactions, which are crucial for the development of fibrosis and weakly represent the derived tissue. Real-time PCR analysis of fibroblasts derived from control and DD samples show that little difference is detectable. 3D cultures of fibroblasts include addition of extracellular matrix that alters the native conditions of these cells. As a way to characterize the fibrotic, proliferative properties of these resection specimens we have developed a 3D culture system, using intact human resections of the nodule part of the cord. The system is based on transwell plates with an attached nitrocellulose membrane that allows contact of the tissue with the medium but not with the plastic, thus, preventing the alteration of the tissue. No collagen gel or other extracellular matrix protein substrate is required. The tissue resection specimens maintain their viability and proliferative properties for 7 days. This is the first "organ" culture system that allows human resection specimens from DD patients to be grown ex vivo and functionally tested, recapitulating the in vivo situation.

  8. Impact of acetaminophen consumption and resistance exercise on extracellular matrix gene expression in human skeletal muscle.

    Science.gov (United States)

    Patel, Shivam H; D'Lugos, Andrew C; Eldon, Erica R; Curtis, Donald; Dickinson, Jared M; Carroll, Chad C

    2017-07-01

    Acetaminophen (APAP) given during chronic exercise reduces skeletal muscle collagen and cross-linking in rats. We propose that the effect of APAP on muscle extracellular matrix (ECM) may, in part, be mediated by dysregulation of the balance between matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs). The purpose of this study was to evaluate the impact of APAP consumption during acute resistance exercise (RE) on several regulators of the ECM in human skeletal muscle. In a double-blinded, placebo-controlled, randomized crossover design, recreationally active men (n = 8, 25 ± 2 yr) performed two trials of knee extension. Placebo (PLA) or APAP (1,000 mg/6 h) was given for 24 h before and immediately following RE. Vastus lateralis biopsies were taken at baseline and 1 and 3 h post-RE. Quantitative RT-PCR was used to determine differences in mRNA expression. MMP-2, type I collagen, and type III collagen mRNA expression was not altered by exercise or APAP (P > 0.05). When compared with PLA, TIMP-1 expression was lower at 1 h post-RE during APAP conditions but greater than PLA at 3 h post-RE (P 0.05). Phosphorylation of ERK1/2 and p38-MAPK increased (P < 0.05) with RE but was not influenced by APAP. Our findings do not support our hypothesis and suggest that short-term APAP consumption before RE has a small impact on the measured ECM molecules in human skeletal muscle following acute RE. Copyright © 2017 the American Physiological Society.

  9. Identification of calprotectin, a calcium binding leukocyte protein, in human dental calculus matrix.

    Science.gov (United States)

    Kido, J; Nishikawa, S; Ishida, H; Yamashita, K; Kitamura, S; Kohri, K; Nagata, T

    1997-05-01

    Calprotectin is a calcium binding protein produced by leukocytes, macrophages and epithelial cells, and its levels in several tissues increase during infections and in many inflamed areas, suggesting that it may be an indicator of inflammatory activity. Osteopontin is a prominent phosphorylated glycoprotein in bone matrix, having calcium binding capacity. Recently, it has been reported that calprotectin and osteopontin are present in urinary stones (pathological mineralized masses in the body), and that these proteins may be involved in their formation. Dental calculus formed by mineralization of dental plaque is an inflammatory factor which may contribute to periodontal disease. It contains many organic components involved in mineralization. We recently found osteopontin molecules in human dental calculus and suggested that the components of its matrix may be similar to those of urinary stones. In this study, we investigated the presence of calprotectin in human dental calculus by immunohistochemical and immunoblotting analyses using a specific antibody for calprotectin. After fixation and demineralization of dental calculi adhered to tooth roots, sections embedded in paraffin were immunoreacted with the antibody for calprotectin and positive immunostaining for calprotectin was observed. Dental calculus proteins were then extracted with EDTA and separated by electrophoresis on 15% polyacrylamide gels. By immunoblotting analysis, 3 or 4 bands were observed at 11, 14.5, 22-25, 28 or 36.5 kDa and these patterns corresponded to those of calprotectin subunits. When non-immune rabbit serum was used instead of calprotectin-specific antibody as a negative control, no immunoreactivity was observed. These findings indicate that calprotectin is associated not only with antibacterial action but also with calcium binding capacity during dental calculus formation.

  10. Degradation and erosion mechanisms of bioresorbable porous acellular vascular grafts: an in vitro investigation.

    Science.gov (United States)

    Gade, Piyusha S; Lee, Keewon; Pfaff, Blaise N; Wang, Yadong; Robertson, Anne M

    2017-07-01

    A fundamental mechanism of in situ tissue regeneration from biodegradable synthetic acellular vascular grafts is the effective interplay between graft degradation, erosion and the production of extracellular matrix. In order to understand this crucial process of graft erosion and degradation, we conducted an in vitro investigation of grafts (n = 4 at days 1, 4, 7, 10 each) exposed to enzymatic degradation. Herein, we provide constitutive relationships for mass loss and mechanical properties based on much-needed experimental data. Furthermore, we formulate a mathematical model to provide a physics-based framework for understanding graft erosion. A novel finding is that despite their porous nature, grafts lost mass exponentially via surface erosion demonstrating a 20% reduction in outer diameter and no significant change in apparent density. A diffusion based, concentration gradient-driven mechanistic model of mass loss through surface erosion was introduced which can be extended to an in vivo setting through the use of two degradation parameters. Furthermore, notably, mechanical properties of degrading grafts did not scale with mass loss. Thus, we introduced a damage function scaling a neo-Hookean model to describe mechanical properties of the degrading graft; a refinement to existing mass-dependent growth and remodelling (G&R) models. This framework can be used to improve accuracy of well-established G&R theories in biomechanics; tools that predict evolving structure-function relationships of neotissues and guide graft design. © 2017 The Author(s).

  11. Acellular matrix scaffold for tissue-engineered intervertebral disc which is closest to the normal three-dimensional structure of the nucleus pulposus%去细胞基质支架构建组织工程椎间盘:最为接近正常髓核三维结构

    Institute of Scientific and Technical Information of China (English)

    谭伟; 吕海; 周初松

    2015-01-01

    背景:目前临床上对椎间盘退变疾病无特效治疗,组织工程技术的发展为它提供了新的治疗思路。目的:综述了去细胞基质支架构建组织工程椎间盘的研究的研究进展。方法:应用计算机检索PubMed、中国知网、万方、中国生物医学文献库数据库2005至2014年文献,检索关键词为“椎间盘退变,去细胞基质,组织工程,支架,髓核,纤维环;Intervertebral disc degeneration, Extracelular matrix,Tissue Engineering,Scaffold,nucleus pulposus,annulus fibrosus”。结果与结论:组织工程椎间盘构建包括3要素:细胞支架、种子细胞及细胞因子,支架是其中的关键。去细胞基质是其中一类重要支架,近年来成为组织工程椎间盘支架的研究热点,可分为去细胞纤维环支架、去细胞髓核支架及一体化椎间盘支架,其中不同支架具有多种制备方法。相对各种人工材料来源及单一天然蛋白成分来源的支架,去细胞基质具有自己显著的优点,抗原性小、生物相容性高、最为接近正常髓核三维结构,可提供细胞生长所需微环境。但去细胞基质做为组织工程椎间盘支架同时仍存在少量不足,亟待进一步改进。%BACKGROUND:At present there is no specific therapy for the treatment of degenerateive disc diseases. The development of tissue engineering technology provides a new therapy idea for it. OBJECTIVE:To review the research progress of acelular matrix scaffold to construct tissue-engineered intervertebral disc. METHODS:A computer-based online search of PubMed, CNKI, Wanfang and Chinese Biomedical Database was performed for relevant articles published between 2005 to 2014 using the keywords of “intervertebral disc degeneration, extracelular matrix, tissue engineering, scaffold, nucleus pulposus, annulus fibrosus” in English and Chinese, respectively. RESULTS AND CONCLUSION:Construction of tissue

  12. Biocompatibility and superiority of lyophilized acellular ligament scaffolds%冻干韧带脱细胞支架材料的生物相容性及优势

    Institute of Scientific and Technical Information of China (English)

    王辉; 陈雄生; 周盛源; 黄俊俊; 蔡弢艺

    2011-01-01

    BACKGROUND: Acellular matrix ligament removes the cellular components within the ligament tissue and reduce the immunogenicity through a variety of acellular ways. Simultaneously, the damage to scaffold structure is mild in the process of decellularization, and it retains the mechanical properties of the extracellular matrix.OBJECTIVE: To verify the biocompatibility and superiority of rabbit patellar tendon acellular scaffold after frozen and lyophilized processing.METHODS: Patellar ligaments were treated with 1% sodium deoxycholate for preparation of acellular ligaments with or without lyophilization. RESULTS AND CONCLUSION: No residual nuclear component was detected in all ligaments. Collagen structure was maintained.No significant differences in elastic modulus and ultimate tensile stress were found between non-lyophilized acellular scaffolds and lyophilized ones. The in vitro cytotoxicity test showed the cells grew well in all groups with or without extracts from lyophilized acellular scaffold. No significant difference was found between the control group and the experiment group. Toxicity symptoms were not obvious.Pyrogen detection experiment showed that no pyrogen was found in the lyophilized acellular scaffold extracts. Percutaneous stimulation test was negative as primary stimulation index was 0. In vivo implantation experiment showed that lyophilized acellular ligament scaffold showed the characteristics of little immunogenicity and light inflammation. Lyophilized acellular ligament scaffold treated with 1% DCA method not only maintains the mechanical characteristics of the non-lyophilized ones, but also has good biological compatibility. Because of its preparation, disinfection, packaging and preservation was easy and convenient, the lyophilized acellular ligament scaffold will be an ideal scaffold for tissue engineering ligament.%背景:韧带脱细胞基质是通过各种脱细胞方法将韧带组织内的细胞成分清除,降低免疫

  13. Dexamethasone-Mediated Activation of Fibronectin Matrix Assembly Reduces Dispersal of Primary Human Glioblastoma Cells.

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    Stephen Shannon

    Full Text Available Despite resection and adjuvant therapy, the 5-year survival for patients with Glioblastoma multiforme (GBM is less than 10%. This poor outcome is largely attributed to rapid tumor growth and early dispersal of cells, factors that contribute to a high recurrence rate and poor prognosis. An understanding of the cellular and molecular machinery that drive growth and dispersal is essential if we are to impact long-term survival. Our previous studies utilizing a series of immortalized GBM cell lines established a functional causation between activation of fibronectin matrix assembly (FNMA, increased tumor cohesion, and decreased dispersal. Activation of FNMA was accomplished by treatment with Dexamethasone (Dex, a drug routinely used to treat brain tumor related edema. Here, we utilize a broad range of qualitative and quantitative assays and the use of a human GBM tissue microarray and freshly-isolated primary human GBM cells grown both as conventional 2D cultures and as 3D spheroids to explore the role of Dex and FNMA in modulating various parameters that can significantly influence tumor cell dispersal. We show that the expression and processing of fibronectin in a human GBM tissue-microarray is variable, with 90% of tumors displaying some abnormality or lack in capacity to secrete fibronectin or assemble it into a matrix. We also show that low-passage primary GBM cells vary in their capacity for FNMA and that Dex treatment reactivates this process. Activation of FNMA effectively "glues" cells together and prevents cells from detaching from the primary mass. Dex treatment also significantly increases the strength of cell-ECM adhesion and decreases motility. The combination of increased cohesion and decreased motility discourages in vitro and ex vivo dispersal. By increasing cell-cell cohesion, Dex also decreases growth rate of 3D spheroids. These effects could all be reversed by an inhibitor of FNMA and by the glucocorticoid receptor antagonist, RU

  14. Risk of Brain Damage Following Pertussis Immunization with Whole-Cell cf Acellular Vaccines

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    J Gordon Millichap

    2004-06-01

    Full Text Available Serious neurological disorders reported following whole-cell (WC in comparison to acellular (AC pertussis vaccines (PV were evaluated by the Genetic Centers of America, Silver Spring, MD.

  15. Changes in extracellular matrix composition regulate cyclooxygenase-2 expression in human mesangial cells.

    Science.gov (United States)

    Alique, Matilde; Calleros, Laura; Luengo, Alicia; Griera, Mercedes; Iñiguez, Miguel Ángel; Punzón, Carmen; Fresno, Manuel; Rodríguez-Puyol, Manuel; Rodríguez-Puyol, Diego

    2011-04-01

    Glomerular diseases are characterized by a sustained synthesis and accumulation of abnormal extracellular matrix proteins, such as collagen type I. The extracellular matrix transmits information to cells through interactions with membrane components, which directly activate many intracellular signaling events. Moreover, accumulating evidence suggests that eicosanoids derived from cyclooxygenase (COX)-2 participate in a number of pathological processes in immune-mediated renal diseases, and it is known that protein kinase B (AKT) may act through different transcription factors in the regulation of the COX-2 promoter. The present results show that progressive accumulation of collagen I in the extracellular medium induces a significant increase of COX-2 expression in human mesangial cells, resulting in an enhancement in PGE(2) production. COX-2 overexpression is due to increased COX-2 mRNA levels. The study of the mechanism implicated in COX-2 upregulation by collagen I showed focal adhesion kinase (FAK) activation. Furthermore, we observed that the activation of the phosphatidylinositol 3-kinase (PI3K)/AKT pathway by collagen I and collagen I-induced COX-2 overexpression was abolished by PI3K and AKT inhibitors. Additionally, we showed that the cAMP response element (CRE) transcription factor is implicated. Finally, we studied COX-2 expression in an animal model, N(G)-nitro-l-arginine methyl ester hypertensive rats. In renal tissue and vascular walls, COX-2 and collagen type I content were upregulated. In summary, our results provide evidence that collagen type I increases COX-2 expression via the FAK/PI3K/AKT/cAMP response element binding protein signaling pathway.

  16. IAPP modulates cellular autophagy, apoptosis, and extracellular matrix metabolism in human intervertebral disc cells

    Science.gov (United States)

    Wu, Xinghuo; Song, Yu; Liu, Wei; Wang, Kun; Gao, Yong; Li, Shuai; Duan, Zhenfeng; Shao, Zengwu; Yang, Shuhua; Yang, Cao

    2017-01-01

    The pathogenic process of intervertebral disc degeneration (IDD) is characterized by imbalance in the extracellular matrix (ECM) metabolism. Nucleus pulposus (NP) cells have important roles in maintaining the proper structure and tissue homeostasis of disc ECM. These cells need adequate supply of glucose and oxygen. Islet amyloid polypeptide (IAPP) exerts its biological effects by regulating glucose metabolism. The purpose of this study was to investigate the expression of IAPP in degenerated IVD tissue, and IAPP modulation of ECM metabolism in human NP cells, especially the crosstalk mechanism between apoptosis and autophagy in these cells. We found that the expression of IAPP and Calcr-RAMP decreased considerably during IDD progression, along with the decrease in the expression of AG, BG, and Col2A1. Induction of IAPP in NP cells by transfection with pLV-IAPP enhanced the synthesis of aggrecan and Col2A1 and attenuated the expression of pro-inflammatory factors, tumor necrosis factor (TNF)-α, and interleukin (IL)-1. Upregulation of IAPP also affected the expression of the catabolic markers—matrix metalloproteinases (MMPs) 3, 9 and 13 and ADAMTS 4 and 5. Downregulation of IAPP by siRNA inhibited the expression of anabolic genes but increased the expression of catabolic genes and inflammatory factors. The expressions of autophagic and apoptotic markers in NP cells transfected with pLV-IAPP were upregulated, including BECLIN1, ATG5, ATG7, LC3 II/I and Bcl-2, while significantly increase in the expression of Bax and Caspase-3 in NP cells transfected with pLV-siIAPP. Mechanistically, PI3K/AKT-mTOR and p38/JNK MAPK signal pathways were involved. We propose that IAPP might play a pivotal role in the development of IDD, by regulating ECM metabolism and controlling the crosstalk between apoptosis and autophagy in NP, thus potentially offering a novel therapeutic approach to the treatment of IDD.

  17. Expression pattern of matrix metalloproteinases in human gynecological cancer cell lines

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    Feix Sonja

    2010-10-01

    Full Text Available Abstract Background Matrix metalloproteinases (MMPs are involved in the degradation of protein components of the extracellular matrix and thus play an important role in tumor invasion and metastasis. Their expression is related to the progression of gynecological cancers (e.g. endometrial, cervical or ovarian carcinoma. In this study we investigated the expression pattern of the 23 MMPs, currently known in humans, in different gynecological cancer cell lines. Methods In total, cell lines from three endometrium carcinomas (Ishikawa, HEC-1-A, AN3 CA, three cervical carcinomas (HeLa, Caski, SiHa, three chorioncarcinomas (JEG, JAR, BeWo, two ovarian cancers (BG-1, OAW-42 and one teratocarcinoma (PA-1 were examined. The expression of MMPs was analyzed by RT-PCR, Western blot and gelatin zymography. Results We demonstrated that the cell lines examined can constitutively express a wide variety of MMPs on mRNA and protein level. While MMP-2, -11, -14 and -24 were widely expressed, no expression was seen for MMP-12, -16, -20, -25, -26, -27 in any of the cell lines. A broad range of 16 MMPs could be found in the PA1 cells and thus this cell line could be used as a positive control for general MMP experiments. While the three cervical cancer cell lines expressed 10-14 different MMPs, the median expression in endometrial and choriocarcinoma cells was 7 different enzymes. The two investigated ovarian cancer cell lines showed a distinctive difference in the number of expressed MMPs (2 vs. 10. Conclusions Ishikawa, Caski, OAW-42 and BeWo cell lines could be the best choice for all future experiments on MMP regulation and their role in endometrial, cervical, ovarian or choriocarcinoma development, whereas the teratocarcinoma cell line PA1 could be used as a positive control for general MMP experiments.

  18. Silicon-based quantum dots induce inflammation in human lung cells and disrupt extracellular matrix homeostasis.

    Science.gov (United States)

    Stan, Miruna-Silvia; Sima, Cornelia; Cinteza, Ludmila Otilia; Dinischiotu, Anca

    2015-08-01

    Quantum dots (QDs) are nanocrystalline semiconductor materials that have been tested for biological applications such as cancer therapy, cellular imaging and drug delivery, despite the serious lack of information of their effects on mammalian cells. The present study aimed to evaluate the potential of Si/SiO2 QDs to induce an inflammatory response in MRC-5 human lung fibroblasts. Cells were exposed to different concentrations of Si/SiO2 QDs (25-200 μg·mL(-1)) for 24, 48, 72 and 96 h. The results obtained showed that uptake of QDs was dependent on biocorona formation and the stability of nanoparticles in various biological media (minimum essential medium without or with 10% fetal bovine serum). The cell membrane damage indicated by the increase in lactate dehydrogenase release after exposure to QDs was dose- and time-dependent. The level of lysosomes increased proportionally with the concentration of QDs, whereas an accumulation of autophagosomes was also observed. Cellular morphology was affected, as shown by the disruption of actin filaments. The enhanced release of nitric oxide and the increase in interleukin-6 and interleukin-8 protein expression suggested that nanoparticles triggered an inflammatory response in MRC-5 cells. QDs decreased the protein expression and enzymatic activity of matrix metalloproteinase (MMP)-2 and MMP-9 and also MMP-1 caseinase activity, whereas the protein levels of MMP-1 and tissue inhibitor of metalloproteinase-1 increased. The present study reveals for the first time that silicon-based QDs are able to generate inflammation in lung cells and cause an imbalance in extracellular matrix turnover through a differential regulation of MMPs and tissue inhibitor of metalloproteinase-1 protein expression.

  19. The human metapneumovirus matrix protein stimulates the inflammatory immune response in vitro.

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    Audrey Bagnaud-Baule

    Full Text Available Each year, during winter months, human Metapneumovirus (hMPV is associated with epidemics of bronchiolitis resulting in the hospitalization of many infants. Bronchiolitis is an acute illness of the lower respiratory tract with a consequent inflammation of the bronchioles. The rapid onset of inflammation suggests the innate immune response may have a role to play in the pathogenesis of this hMPV infection. Since, the matrix protein is one of the most abundant proteins in the Paramyxoviridae family virion, we hypothesized that the inflammatory modulation observed in hMPV infected patients may be partly associated with the matrix protein (M-hMPV response. By western blot analysis, we detected a soluble form of M-hMPV released from hMPV infected cell as well as from M-hMPV transfected HEK 293T cells suggesting that M-hMPV may be directly in contact with antigen presenting cells (APCs during the course of infection. Moreover, flow cytometry and confocal microscopy allowed determining that M-hMPV was taken up by dendritic cells (moDCs and macrophages inducing their activation. Furthermore, these moDCs enter into a maturation process inducing the secretion of a broad range of inflammatory cytokines when exposed to M-hMPV. Additionally, M-hMPV activated DCs were shown to stimulate IL-2 and IFN-γ production by allogeneic T lymphocytes. This M-hMPV-mediated activation and antigen presentation of APCs may in part explain the marked inflammatory immune response observed in pathology induced by hMPV in patients.

  20. CHANGES OF NUCLEAR MATRIX PROTEIN AND ITS RELATIONSHIP WITH c-erbB-2 IN HUMAN COLON ADENOCARCINOMA

    Institute of Scientific and Technical Information of China (English)

    WANG Ya-lan; GAO Jing; LI Yuan-yuan

    2005-01-01

    Objective: Nuclear matrix protein is tissue, cell-type specific, and tumor-relative. It plays an important role in the regulation of intranuclear processes. Some researches also showed that a c-erbB-2 promoter-specific DNA-binding nuclear matrix protein is present only in malignant human breast tissues and induces mitogenesis and cell surface expression of the c-erbB-2 protein in resting NIH/3T3 cells. But it is not clear that how it in colon adenocarcinomas. Methods:Two-dimensional gel electrophoretic method was used for NMP identification and immunohistochemistry was used for c-erbB-2 detection in 12 cases of colon adenocarcinomas and matched adjacent normal colon tissues. Results: 5 different nuclear matrix proteins (named C1-C5) were identified in 12 colon adenocarcinoma specimens, but not in the matched adjacent normal colon tissues; 3 nuclear matrix proteins (named N1-N3) were identified in all 12 matched adjacent normal colon tissues, but not in colon adenocarcinoma specimens. A nuclear matrix protein (named N4) was detected in all of 9moderated-well differentiated adenocarcinomas and all 12 matched adjacent normal colon tissues, but not in 3poor-differentiated adenocarcinomas. All of the 10 colon adenocarcinomas which had the nuclear matrix protein C4 were c-erbB-2 expression positive. Conclusion: The data suggest that there are specific nuclear matrix proteins in colon adenocarcinomas and its subtypes, which maybe valuable to serve as markers of colon adenocarcinomas in future. Nuclear matrix protein C4 probably is a c-erbB-2 promotor-specific nuclear matrix protein in colon adenocarcinomas, and may induce the expression of c-erbB-2.

  1. CADM1 controls actin cytoskeleton assembly and regulates extracellular matrix adhesion in human mast cells.

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    Elena P Moiseeva

    Full Text Available CADM1 is a major receptor for the adhesion of mast cells (MCs to fibroblasts, human airway smooth muscle cells (HASMCs and neurons. It also regulates E-cadherin and alpha6beta4 integrin in other cell types. Here we investigated a role for CADM1 in MC adhesion to both cells and extracellular matrix (ECM. Downregulation of CADM1 in the human MC line HMC-1 resulted not only in reduced adhesion to HASMCs, but also reduced adhesion to their ECM. Time-course studies in the presence of EDTA to inhibit integrins demonstrated that CADM1 provided fast initial adhesion to HASMCs and assisted with slower adhesion to ECM. CADM1 downregulation, but not antibody-dependent CADM1 inhibition, reduced MC adhesion to ECM, suggesting indirect regulation of ECM adhesion. To investigate potential mechanisms, phosphotyrosine signalling and polymerisation of actin filaments, essential for integrin-mediated adhesion, were examined. Modulation of CADM1 expression positively correlated with surface KIT levels and polymerisation of cortical F-actin in HMC-1 cells. It also influenced phosphotyrosine signalling and KIT tyrosine autophosphorylation. CADM1 accounted for 46% of surface KIT levels and 31% of F-actin in HMC-1 cells. CADM1 downregulation resulted in elongation of cortical actin filaments in both HMC-1 cells and human lung MCs and increased cell rigidity of HMC-1 cells. Collectively these data suggest that CADM1 is a key adhesion receptor, which regulates MC net adhesion, both directly through CADM1-dependent adhesion, and indirectly through the regulation of other adhesion receptors. The latter is likely to occur via docking of KIT and polymerisation of cortical F-actin. Here we propose a stepwise model of adhesion with CADM1 as a driving force for net MC adhesion.

  2. Generation of tissue-engineered small intestine using embryonic stem cell-derived human intestinal organoids.

    Science.gov (United States)

    Finkbeiner, Stacy R; Freeman, Jennifer J; Wieck, Minna M; El-Nachef, Wael; Altheim, Christopher H; Tsai, Yu-Hwai; Huang, Sha; Dyal, Rachel; White, Eric S; Grikscheit, Tracy C; Teitelbaum, Daniel H; Spence, Jason R

    2015-10-12

    Short bowel syndrome (SBS) is characterized by poor nutrient absorption due to a deficit of healthy intestine. Current treatment practices rely on providing supportive medical therapy with parenteral nutrition; while life saving, such interventions are not curative and are still associated with significant co-morbidities. As approaches to lengthen remaining intestinal tissue have been met with only limited success and intestinal transplants have poor survival outcomes, new approaches to treating SBS are necessary. Human intestine derived from embryonic stem cells (hESCs) or induced pluripotent stem cells (iPSCs), called human intestinal organoids (HIOs), have the potential to offer a personalized and scalable source of intestine for regenerative therapies. However, given that HIOs are small three-dimensional structures grown in vitro, methods to generate usable HIO-derived constructs are needed. We investigated the ability of hESCs or HIOs to populate acellular porcine intestinal matrices and artificial polyglycolic/poly L lactic acid (PGA/PLLA) scaffolds, and examined the ability of matrix/scaffolds to thrive when transplanted in vivo. Our results demonstrate that the acellular matrix alone is not sufficient to instruct hESC differentiation towards an endodermal or intestinal fate. We observed that while HIOs reseed acellular porcine matrices in vitro, the HIO-reseeded matrices do not thrive when transplanted in vivo. In contrast, HIO-seeded PGA/PLLA scaffolds thrive in vivo and develop into tissue that looks nearly identical to adult human intestinal tissue. Our results suggest that HIO-seeded PGA/PLLA scaffolds are a promising avenue for developing the mucosal component of tissue engineered human small intestine, which need to be explored further to develop them into fully functional tissue.

  3. Generation of tissue-engineered small intestine using embryonic stem cell-derived human intestinal organoids

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    Stacy R. Finkbeiner

    2015-11-01

    Full Text Available Short bowel syndrome (SBS is characterized by poor nutrient absorption due to a deficit of healthy intestine. Current treatment practices rely on providing supportive medical therapy with parenteral nutrition; while life saving, such interventions are not curative and are still associated with significant co-morbidities. As approaches to lengthen remaining intestinal tissue have been met with only limited success and intestinal transplants have poor survival outcomes, new approaches to treating SBS are necessary. Human intestine derived from embryonic stem cells (hESCs or induced pluripotent stem cells (iPSCs, called human intestinal organoids (HIOs, have the potential to offer a personalized and scalable source of intestine for regenerative therapies. However, given that HIOs are small three-dimensional structures grown in vitro, methods to generate usable HIO-derived constructs are needed. We investigated the ability of hESCs or HIOs to populate acellular porcine intestinal matrices and artificial polyglycolic/poly L lactic acid (PGA/PLLA scaffolds, and examined the ability of matrix/scaffolds to thrive when transplanted in vivo. Our results demonstrate that the acellular matrix alone is not sufficient to instruct hESC differentiation towards an endodermal or intestinal fate. We observed that while HIOs reseed acellular porcine matrices in vitro, the HIO-reseeded matrices do not thrive when transplanted in vivo. In contrast, HIO-seeded PGA/PLLA scaffolds thrive in vivo and develop into tissue that looks nearly identical to adult human intestinal tissue. Our results suggest that HIO-seeded PGA/PLLA scaffolds are a promising avenue for developing the mucosal component of tissue engineered human small intestine, which need to be explored further to develop them into fully functional tissue.

  4. [SPREADING OF TISSUE SPHEROIDS FROM PRIMARY HUMAN FIBROBLASTS ON THE SURFACE OF MICROFIBROUS ELECTROSPUN POLYURETHANE MATRIX (A scanning electron microscopic study)].

    Science.gov (United States)

    Kudan, Ye V; Pereira, F D A S; Parfenov, V A; Kasyanov, V A; Khesuani, Yu D; Bulanova, Ye A; Mironov, V A

    2015-01-01

    Tissue spheroids biofabricated from primary human fibroblasts using non-adhesive agarose forms, were placed by 3D bioprinter on the surface of microfibrous electrospun matrix. It was demonstrated that tissue spheroids attached to the surface of matrix during several hours and then gradually spread for several days which indicates high level of biocompatibiity of electrospun microfibrous polyurethane matrix. During this activity, human fibroblasts used processes of leading cell borders for initial step of attachment to matrix filaments. Tissue constructions formed during spreading of tissue spheroids on the surface of electrospun microfibrous polyurethane matrix seem to be a perspective technology platform for development of new methods of biofabrication and 3D bioprinting.

  5. Proliferation and extracellular matrix synthesis of smooth muscle cells cultured from human coronary atherosclerotic and restenotic lesions

    NARCIS (Netherlands)

    D.C. MacLeod (Donald); B.H. Strauss (Bradley); J. Escaned (Javier); V.A.W.M. Umans (Victor); R-J. van Suylen (Robert-Jan); A. Verkerk (Anton); P.J. de Feyter (Pim); P.W.J.C. Serruys (Patrick); M. de Jong (Marcel)

    1994-01-01

    textabstractOBJECTIVES. The purpose of this study was to examine the proliferative capacity and extracellular matrix synthesis of human coronary plaque cells in vitro. BACKGROUND. Common to both primary atherosclerosis and restenosis are vascular smooth muscle cell proliferation and production of ex

  6. The effect of oxygen tension on human articular chondrocyte matrix synthesis: integration of experimental and computational approaches.

    Science.gov (United States)

    Li, S; Oreffo, R O C; Sengers, B G; Tare, R S

    2014-09-01

    Significant oxygen gradients occur within tissue engineered cartilaginous constructs. Although oxygen tension is an important limiting parameter in the development of new cartilage matrix, its precise role in matrix formation by chondrocytes remains controversial, primarily due to discrepancies in the experimental setup applied in different studies. In this study, the specific effects of oxygen tension on the synthesis of cartilaginous matrix by human articular chondrocytes were studied using a combined experimental-computational approach in a "scaffold-free" 3D pellet culture model. Key parameters including cellular oxygen uptake rate were determined experimentally and used in conjunction with a mathematical model to estimate oxygen tension profiles in 21-day cartilaginous pellets. A threshold oxygen tension (pO2 ≈ 8% atmospheric pressure) for human articular chondrocytes was estimated from these inferred oxygen profiles and histological analysis of pellet sections. Human articular chondrocytes that experienced oxygen tension below this threshold demonstrated enhanced proteoglycan deposition. Conversely, oxygen tension higher than the threshold favored collagen synthesis. This study has demonstrated a close relationship between oxygen tension and matrix synthesis by human articular chondrocytes in a "scaffold-free" 3D pellet culture model, providing valuable insight into the understanding and optimization of cartilage bioengineering approaches.

  7. Self-similarity matrix based slow-time feature extraction for human target in high-resolution radar

    NARCIS (Netherlands)

    He, Y.; Aubry, P.; Le Chevalier, F.; Yarovoy, A.

    2014-01-01

    A new approach is proposed to extract the slow-time feature of human motion in high-resolution radars. The approach is based on the self-similarity matrix (SSM) of the radar signals. The Mutual Information is used as a measure of similarity. The SSMs of different radar signals (high-resolution range

  8. Human breast cancer cell-mediated bone collagen degradation requires plasminogen activation and matrix metalloproteinase activity

    Directory of Open Access Journals (Sweden)

    Hill Peter A

    2005-02-01

    Full Text Available Abstract Background Breast cancer cells frequently metastasize to the skeleton and induce extensive bone destruction. Cancer cells produce proteinases, including matrix metalloproteinases (MMPs and the plasminogen activator system (PAS which promote invasion of extracellular matrices, but whether these proteinases degrade bone matrix is unclear. To characterize the role that breast cancer cell proteinases play in bone degradation we compared the effects of three human breast cancer cell lines, MDA-MB-231, ZR-75-1 and MCF-7 with those of a normal breast epithelial cell line, HME. The cell lines were cultured atop radiolabelled matrices of either mineralized or non-mineralized bone or type I collagen, the principal organic constituent of bone. Results The 3 breast cancer cell lines all produced significant degradation of the 3 collagenous extracellular matrices (ECMs whilst the normal breast cell line was without effect. Breast cancer cells displayed an absolute requirement for serum to dissolve collagen. Degradation of collagen was abolished in plasminogen-depleted serum and could be restored by the addition of exogenous plasminogen. Localization of plasmin activity to the cell surface was critical for the degradation process as aprotinin, but not α2 antiplasmin, prevented collagen dissolution. During ECM degradation breast cancer cell lines expressed urokinase-type plasminogen activator (u-PA and uPA receptor, and MMPs-1, -3, -9,-13, and -14. The normal breast epithelial cell line expressed low levels of MMPs-1, and -3, uPA and uPA receptor. Inhibitors of both the PAS (aprotinin and PA inhibitor-1 and MMPs (CT1166 and tisue inhibitor of metalloproteinase blocked collagen degradation, demonstrating the requirement of both plasminogen activation and MMP activity for degradation. The activation of MMP-13 in human breast cancer cells was prevented by plasminogen activator inhibitor-1 but not by tissue inhibitor of metalloproteinase-1, suggesting

  9. Knockdown of asporin affects transforming growth factor-β1-induced matrix synthesis in human intervertebral annulus cells

    Directory of Open Access Journals (Sweden)

    Xu Jiang

    2016-10-01

    Conclusion: Our results have verified a functional feedback loop between TGF-β1 and asporin in human intervertebral annulus cells indicating that TGF-β1-induced annulus matrix biosynthesis can be significantly upregulated by knockdown of asporin. Therefore, asporin could be a potential new therapeutic target and inhibition of asporin could be adopted to enhance the anabolic effect of TGF-β1 in human intervertebral annulus cells in degenerative IVD diseases.

  10. The 5‘—flanking cis—acting elements of the human ε—globin gene associates with the nuclear matrix and binds to the nuclear matrix proteins

    Institute of Scientific and Technical Information of China (English)

    YANZHIJIANG; RUOLANQIAN

    1998-01-01

    The nuclear matrix attachment regions(MARs) and the binding nuclear matrix proteins in the 5'-flanking cisacting elements of the human ε-globin gene have been examined.Using in vitro DNA-matrix binding assay,it has been shown that the positive stage-specific regulatory element (ε-PREII,-446bp- -419bp) upstream of this gene could specifically associate with the nuclear matrix from K562 cells,indicating that ε-PREII may be an erythroidspecific facultative MAR.In gel mobility shift assay and Southwestern blotting assay,an erythroid-specific nuclear matrix protein (ε-NMPk) in K562 cells has been revealed to bind to this positive regulatory element (ε-PREII).Furthermore,we demonstrated that the silencer (-392bp- -177bp) upstream of the human ε-globin gene could associate with the nuclear matrices from K562,HEL and Raji cells.In addition,the nuclear matrix proteins prepared from these three cell lines could also bind to this silencer,suggesting that this silencer element might be a constitutive nuclear matrix attachment region(constitutive MAR).Our results demonstrated that the nuclear matrix and nuclear matrix proteins might play an important role in the regulation of the human ε-globin gene expression.

  11. Three-Dimensional In Vitro Skin and Skin Cancer Models Based on Human Fibroblast-Derived Matrix.

    Science.gov (United States)

    Berning, Manuel; Prätzel-Wunder, Silke; Bickenbach, Jackie R; Boukamp, Petra

    2015-09-01

    Three-dimensional in vitro skin and skin cancer models help to dissect epidermal-dermal and tumor-stroma interactions. In the model presented here, normal human dermal fibroblasts isolated from adult skin self-assembled into dermal equivalents with their specific fibroblast-derived matrix (fdmDE) over 4 weeks. The fdmDE represented a complex human extracellular matrix that was stabilized by its own heterogeneous collagen fiber meshwork, largely resembling a human dermal in vivo architecture. Complemented with normal human epidermal keratinocytes, the skin equivalent (fdmSE) thereof favored the establishment of a well-stratified and differentiated epidermis and importantly allowed epidermal regeneration in vitro for at least 24 weeks. Moreover, the fdmDE could be used to study the features of cutaneous skin cancer. Complementing fdmDE with HaCaT cells in different stages of malignancy or tumor-derived cutaneous squamous cell carcinoma cell lines, the resulting skin cancer equivalents (fdmSCEs) recapitulated the respective degree of tumorigenicity. In addition, the fdmSCE invasion phenotypes correlated with their individual degree of tissue organization, disturbance in basement membrane organization, and presence of matrix metalloproteinases. Together, fdmDE-based models are well suited for long-term regeneration of normal human epidermis and, as they recapitulate tumor-specific growth, differentiation, and invasion profiles of cutaneous skin cancer cells, also provide an excellent human in vitro skin cancer model.

  12. Pattern secretion of matrix Metalloproteinases and their biological tissue inhibitors by human glomerular mesangial cells in culture

    Directory of Open Access Journals (Sweden)

    "Hosseini R

    2001-08-01

    Full Text Available The glomerular mesangial cells (GMC play a central role in the synthesis and turnover of the glomerular mesangial matrix. The breakdown of the matrix likely depends on the balance between of a variety of proteinases including matrix metalloproteinases and their biological inhibitors secreted by the GMC, and any disturbance in the balance may result in appearance of various pathological states such as glomerulosclerosis. We therefore studied pattern secretion of matrix metalloproteinases (MMPs, MMP-1, MMP-2, MMP-3, MMP-9 and their biological tissue inhibitor of matrix metalloproteinases (TIMPs, TIMP-1 and TIMP-2 by cultured human GMC. We also measured MMP-1/TIMP-1 complex level in the cell culture supernatants. For this purpose, the GMC were incubated under serum-free conditions with medium (RPMI-1640 alone or in combination with TNF-α (30 ng/ml or phorbol myristate acetate (PMA (50 ng/ml for exactly 24, 48 and 72 hours. The above parameters were assayed by established ELISA techniques. Our results showed that the lowest and largest secretions were related to MMP-9 and MMP-2, respectively. The results indicated that the MMPs and TIMPs secretion were increased by TNF-α (MMP-1, MMP-2, TIMP-1 and TIMP-2 and PMA (MMP-2, TIMP-1 and TIMP-2, significantly (P<0.05. These results suggest that the GMC can synthesis and release various MMPs and their inhibitors (TIMPs that, in part, control turnover of extracellular matrix proteins.

  13. Enhanced expression of two discrete isoforms of matrix metalloproteinase-2 in experimental and human diabetic nephropathy

    Science.gov (United States)

    Bae, Sun Sik; Lee, Min Young; Rhee, Harin; Kim, Il Young; Seong, Eun Young; Lee, Dong Won; Lee, Soo Bong; Kwak, Ihm Soo; Lovett, David H.

    2017-01-01

    Background We recently reported on the enhanced expression of two isoforms of matrix metalloproteinase-2 (MMP-2) in human renal transplantation delayed graft function. These consist of the conventional secreted, full length MMP-2 isoform (FL-MMP-2) and a novel intracellular N-Terminal Truncated isoform (NTT-MMP-2) generated by oxidative stress-mediated activation of an alternate promoter in the MMP-2 first intron. Here we evaluated the effect of hyperglycemia and diabetes mellitus on the in vitro and in vivo expression of the two MMP-2 isoforms. Methods We quantified the abundance of the FL-MMP-2 and NTT-MMP-2 transcripts by qPCR in HK2 cells cultured in high glucose or 4-hydroxy-2-hexenal (HHE) and tested the effects of the NF-κB inhibitor pyrrolidine dithiocarbamate (PDTC). The streptozotocin (STZ) murine model of Type I diabetes mellitus and renal biopsies of human diabetic nephropathy were used in this study. Results Both isoforms of MMP-2 in HK2 cells were upregulated by culture in high glucose or with HHE. PDTC treatment did not suppress high glucose-mediated FL-MMP-2 expression but potently inhibited NTT-MMP-2 expression. With STZ-treated mice, renal cortical expression of both isoforms was increased (FL-MMP-2, 1.8-fold; NTT-MMP-2, greater than 7-fold). Isoform-specific immunohistochemical staining revealed low, but detectable levels of the FL-MMP-2 isoform in controls, while NTT-MMP-2 was not detected. While there was a modest increase in tubular epithelial cell staining for FL-MMP-2 in STZ-treated mice, NTT-MMP-2 was intensely expressed in a basolateral pattern. FL-MMP-2 and NTT-MMP-2 isoform expression as quantified by qPCR were both significantly elevated in renal biopsies of human diabetic nephropathy (12-fold and 3-fold, respectively). Conclusions The expression of both isoforms of MMP-2 was enhanced in an experimental model of diabetic nephropathy and in human diabetic nephropathy. Selective MMP-2 isoform inhibition could offer a novel approach for

  14. A new synthetic matrix metalloproteinase inhibitor reduces human mesenchymal stem cell adipogenesis

    Science.gov (United States)

    Bosco, Dale B.; Roycik, Mark D.; Jin, Yonghao; Schwartz, Martin A.; Lively, Ty J.; Zorio, Diego A. R.

    2017-01-01

    Development of adipose tissue requires the differentiation of less specialized cells, such as human mesenchymal stem cells (hMSCs), into adipocytes. Since matrix metalloproteinases (MMPs) play critical roles in the cell differentiation process, we conducted investigations to determine if a novel mercaptosulfonamide-based MMP inhibitor (MMPI), YHJ-7-52, could affect hMSC adipogenic differentiation and lipid accumulation. Enzyme inhibition assays, adipogenic differentiation experiments, and quantitative PCR methods were employed to characterize this inhibitor and determine its effect upon adipogenesis. YHJ-7-52 reduced lipid accumulation in differentiated cells by comparable amounts as a potent hydroxamate MMPI, GM6001. However, YHJ-7-82, a non-inhibitory structural analog of YHJ-7-52, in which the zinc-binding thiol group is replaced by a hydroxyl group, had no effect on adipogenesis. The two MMPIs (YHJ-7-52 and GM6001) were also as effective in reducing lipid accumulation in differentiated cells as T0070907, an antagonist of peroxisome-proliferator activated receptor gamma (PPAR-gamma), at a similar concentration. PPAR-gamma is a typical adipogenic marker and a key regulatory protein for the transition of preadiopocyte to adipocyte. Moreover, MMP inhibition was able to suppress lipid accumulation in cells co-treated with Troglitazone, a PPAR-gamma agonist. Our results indicate that MMP inhibitors may be used as molecular tools for adipogenesis and obesity treatment research. PMID:28234995

  15. Sensitive immunoassay of human chorionic gonadotrophin based on multi-walled carbon nanotube-chitosan matrix.

    Science.gov (United States)

    Li, Na; Yuan, Ruo; Chai, Yaqin; Chen, Shihong; An, Haizhen

    2008-10-01

    A novel amperometric immunosensor for human chorionic gonadotropin (HCG) assay has been fabricated through incorporating toluidine blue (TB) and hemoglobin (Hb) on the multiwall carbon nanotube (MWNT)-chitosan (CS) modified glassy carbon electrode, followed by electrostatic adsorption of a conducting gold nanoparticles (nanogold) film as sensing interface. The MWNT-CS matrix provided a congenial microenvironment for the immobilization of biomolecules and promoted the electron transfer to enhance the sensitivity of the immunosensor. Due to the strong electrocatalytic properties of Hb and MWNT toward H(2)O(2), the Hb and MWNT significantly amplified the current signal of the antigen-antibody reaction. The immobilized toluidine blue as an electron transfer mediator exhibited excellent electrochemical redox property. After the immunosensor was incubated with HCG solution, the access of activity center of the Hb to toluidine blue was partly inhibited, which leaded to a linear decrease in the catalytic efficiency of the Hb to the oxidation of immobilized toluidine blue by H(2)O(2) over HCG concentration ranges from 0.8 to 500 mIU/mL. Under optimal condition, the detection limit for the HCG immunoassay was 0.3 mIU/mL estimated at a signal-to-noise ratio of 3. Moreover, the proposed immunosensor displayed a satisfactory stability and reproducibility.

  16. Increased expression of the matrix metalloproteinase 2 in differentiating Tera 2 human embryonal carcinoma cells.

    Science.gov (United States)

    Tienari, J; Pertovaara, L; Saksela, O; Lehtonen, E; Vartio, T

    1994-01-15

    Secretion of proteolytic enzymes by cells has been implicated in tissue remodeling during embryonic development as well as in invasive neoplastic diseases. We studied the regulation of type-IV-collagenase activity in Tera 2 human embryonal carcinoma cells, which in the undifferentiated state proliferate rapidly and are tumorigenic. The undifferentiated cells produced relatively low levels of matrix-metalloproteinase-2 (MMP-2) activity. This activity was not markedly affected by exogenous basic fibroblast growth factor (bFGF) or 12-O-tetradecanoyl-phorbol-13-acetate (TPA), even though the plasminogen activator activity of the cells was increased by these agents. Tera 2 cells can be induced by retinoic acid to differentiate into quiescent cells, of which many express neuronal characteristics. The type-IV-collagenase activity of the cells increased markedly during the differentiation. This increase was mainly due to increased expression of MMP-2. Expression of tissue inhibitors of metalloproteinases (TIMP-1 and TIMP-2) was not markedly affected by the differentiation of Tera 2 cells. The results show that in the Tera 2 cell system, increased expression of MMP-2 is characteristic of the differentiated derivatives. This is in contrast with many other model systems, where increased type-IV-collagenase activity is associated with the malignant phenotype. This pattern of regulation may reflect the facts that Tera 2 cells resemble early embryonic cells and that their differentiation mimics related cell-differentiation processes in the developing embryo.

  17. Inhibition of matrix metalloproteinases expression in human dental pulp cells by all-trans retinoic acid

    Institute of Scientific and Technical Information of China (English)

    Jin Man Kim; Sang Wook Kang; Su-Mi Shin; Duck Su Kim; Kyong-Kyu Choi; Eun-Cheol Kim; Sun-Young Kim

    2014-01-01

    All-trans retinoic acid (ATRA) inhibits matrix metalloproteinase (MMP)-2 and MMP-9 in synovial fibroblasts, skin fibroblasts, bronchoalveolar lavage cells and cancer cells, but activates MMP-9 in neuroblast and leukemia cells. Very little is known regarding whether ATRA can activate or inhibit MMPs in human dental pulp cells (HDPCs). The purpose of this study was to determine the effects of ATRA on the production and secretion of MMP-2 and-9 in HDPCs. The productions and messenger RNA (mRNA) expressions of MMP-2 and-9 were accessed by gelatin zymography and real-time polymerase chain reaction (PCR), respectively. ATRA was found to decrease MMP-2 level in a dose-dependent manner. Significant reduction in MMP-2 mRNA expression was also observed in HDPCs treated with 25 mmol?L21 ATRA. However, HDPCs treated with ATRA had no effect on the pattern of MMP-9 produced or secreted in either cell extracts or conditioned medium fractions. Taken together, ATRA had an inhibitory effect on MMP-2 expression in HDPCs, which suggests that ATRA could be a candidate as a medicament which could control the inflammation of pulp tissue in vital pulp therapy and regenerative endodontics.

  18. Biocompatibility of pure titanium modified by human endothelial cell-derived extracellular matrix

    Energy Technology Data Exchange (ETDEWEB)

    Xue Xiaoqing [Key Laboratory of Advanced Technology for Materials of Education Ministry, Southwest Jiaotong University, Chengdu 610031 (China); Wang Jin, E-mail: jinxxwang@263.net [Key Laboratory of Advanced Technology for Materials of Education Ministry, Southwest Jiaotong University, Chengdu 610031 (China); Zhu Ying; Tu Qiufen; Huang Nan [Key Laboratory of Advanced Technology for Materials of Education Ministry, Southwest Jiaotong University, Chengdu 610031 (China)

    2010-04-01

    Extracellular matrix (ECM) used to modify biomaterial surface is a promising method for improving cardiovascular material hemocompatibility. In the present work, human umbilical vein endothelial cells (HUVECs) are cultured and native ECM is obtained on pure titanium surface. Fourier infrared spectrum (FTIR) test proves the existence of amide I and amide II band on the modified titanium surface. X-ray photoelectron spectroscopy (XPS) further confirms the chemical composition and binding types of the ECM proteins on the titanium substrate. The results of light microscopy and atomic force microscopy (AFM) exhibit the morphology of HUVEC derived ECM. There are higher water contact angles on the ECM modified samples. Furthermore, some ECM components, including fibronectin (FN), laminin (LN) and type IV collagen (IV-COL) are presented on ECM-covered titanium surface by immunofluorescence staining. The biological behavior of cultured HUVECs and adherent platelets on different samples are investigated by in vitro HUVECs culture and platelet adhesion. Cells exhibit better morphology and their proliferation ability greatly improve on the ECM-covered titanium. At the same time, the platelet adhesion and spreading are inhibited on ECM-covered titanium surface. These investigations demonstrate that ECM produced by HUVECs cannot only improve adhesion and proliferation ability of endothelial cell but also inhibit adhesion and activation of platelets. Thus, the approach described here may provide a basis for preparation of modified surface in cardiovascular implants application.

  19. Urine proteomes of healthy aging humans reveal extracellular matrix (ECM) alterations and immune system dysfunction.

    Science.gov (United States)

    Bakun, M; Senatorski, G; Rubel, T; Lukasik, A; Zielenkiewicz, P; Dadlez, M; Paczek, L

    2014-02-01

    Aging is a complex physiological process that poses considerable conundrums to rapidly aging societies. For example, the risk of dying from cardiovascular diseases and/or cancer steadily declines for people after their 60s, and other causes of death predominate for seniors older than 80 years of age. Thus, physiological aging presents numerous unanswered questions, particularly with regard to changing metabolic patterns. Urine proteomics analysis is becoming a non-invasive and reproducible diagnostic method. We investigated the urine proteomes in healthy elderly people to determine which metabolic processes were weakened or strengthened in aging humans. Urine samples from 37 healthy volunteers aged 19-90 years (19 men, 18 women) were analyzed for protein expression by liquid chromatography-tandem mass spectrometry. This generated a list of 19 proteins that were differentially expressed in different age groups (young, intermediate, and old age). In particular, the oldest group showed protein changes reflective of altered extracellular matrix turnover and declining immune function, in which changes corresponded to reported changes in cardiovascular tissue remodeling and immune disorders in the elderly. Thus, urinary proteome changes in the elderly appear to reflect the physiological processes of aging and are particularly clearly represented in the circulatory and immune systems. Detailed identification of "protein trails" creates a more global picture of metabolic changes that occur in the elderly.

  20. In vitro evaluation of the interactions between human corneal endothelial cells and extracellular matrix proteins.

    Science.gov (United States)

    Choi, Jin San; Kim, Eun Young; Kim, Min Jeong; Giegengack, Matthew; Khan, Faraaz A; Khang, Gilson; Soker, Shay

    2013-02-01

    The corneal endothelium is the innermost cell layer of the cornea and rests on Descemet's membrane consisting of various extracellular matrix (ECM) proteins which can directly affect the cellular behaviors such as cell adhesion, proliferation, polarity, morphogenesis and function. The objective of this study was to investigate the interactions between the ECM environment and human corneal endothelial cells (HCECs), with the ultimate goal to improve cell proliferation and function in vitro. To evaluate the interaction of HCECs with ECM proteins, cells were seeded on ECM-coated tissue culture dishes, including collagen type I (COL I), collagen type IV (COL IV), fibronectin (FN), FNC coating mix (FNC) and laminin (LM). Cell adhesion and proliferation of HCECs on each substratum and expression of CEC markers were studied. The results showed that HCECs plated on the COL I, COL IV, FN and FNC-coated plates had enhanced cell adhesion initially; the number for COL I, COL IV, FN and FNC was significantly higher than the control (P < 0.05). In addition, cells grown on ECM protein-coated dishes showed more compact cellular morphology and CEC marker expression compared to cells seeded on uncoated dishes. Collectively, our results suggest that an adequate ECM protein combination can provide a long-term culture environment for HCECs for corneal endothelium transplantation.

  1. Multiplex matrix network analysis of protein complexes in the human TCR signalosome.

    Science.gov (United States)

    Smith, Stephen E P; Neier, Steven C; Reed, Brendan K; Davis, Tessa R; Sinnwell, Jason P; Eckel-Passow, Jeanette E; Sciallis, Gabriel F; Wieland, Carilyn N; Torgerson, Rochelle R; Gil, Diana; Neuhauser, Claudia; Schrum, Adam G

    2016-08-02

    Multiprotein complexes transduce cellular signals through extensive interaction networks, but the ability to analyze these networks in cells from small clinical biopsies is limited. To address this, we applied an adaptable multiplex matrix system to physiologically relevant signaling protein complexes isolated from a cell line or from human patient samples. Focusing on the proximal T cell receptor (TCR) signalosome, we assessed 210 pairs of PiSCES (proteins in shared complexes detected by exposed surface epitopes). Upon stimulation of Jurkat cells with superantigen-loaded antigen-presenting cells, this system produced high-dimensional data that enabled visualization of network activity. A comprehensive analysis platform generated PiSCES biosignatures by applying unsupervised hierarchical clustering, principal component analysis, an adaptive nonparametric with empirical cutoff analysis, and weighted correlation network analysis. We generated PiSCES biosignatures from 4-mm skin punch biopsies from control patients or patients with the autoimmune skin disease alopecia areata. This analysis distinguished disease patients from the controls, detected enhanced basal TCR signaling in the autoimmune patients, and identified a potential signaling network signature that may be indicative of disease. Thus, generation of PiSCES biosignatures represents an approach that can provide information about the activity of protein signaling networks in samples including low-abundance primary cells from clinical biopsies.

  2. Structure and self-assembly of the calcium binding matrix protein of human metapneumovirus.

    Science.gov (United States)

    Leyrat, Cedric; Renner, Max; Harlos, Karl; Huiskonen, Juha T; Grimes, Jonathan M

    2014-01-07

    The matrix protein (M) of paramyxoviruses plays a key role in determining virion morphology by directing viral assembly and budding. Here, we report the crystal structure of the human metapneumovirus M at 2.8 Å resolution in its native dimeric state. The structure reveals the presence of a high-affinity Ca²⁺ binding site. Molecular dynamics simulations (MDS) predict a secondary lower-affinity site that correlates well with data from fluorescence-based thermal shift assays. By combining small-angle X-ray scattering with MDS and ensemble analysis, we captured the structure and dynamics of M in solution. Our analysis reveals a large positively charged patch on the protein surface that is involved in membrane interaction. Structural analysis of DOPC-induced polymerization of M into helical filaments using electron microscopy leads to a model of M self-assembly. The conservation of the Ca²⁺ binding sites suggests a role for calcium in the replication and morphogenesis of pneumoviruses.

  3. Distribution volumes of macromolecules in human ovarian and endometrial cancers--effects of extracellular matrix structure.

    Science.gov (United States)

    Haslene-Hox, Hanne; Oveland, Eystein; Woie, Kathrine; Salvesen, Helga B; Tenstad, Olav; Wiig, Helge

    2015-01-01

    Elements of the extracellular matrix (ECM), notably collagen and glucosaminoglycans, will restrict part of the space available for soluble macromolecules simply because the molecules cannot occupy the same space. This phenomenon may influence macromolecular drug uptake. To study the influence of steric and charge effects of the ECM on the distribution volumes of macromolecules in human healthy and malignant gynecologic tissues we used as probes 15 abundant plasma proteins quantified by high-resolution mass spectrometry. The available distribution volume (VA) of albumin was increased in ovarian carcinoma compared with healthy ovarian tissue. Furthermore, VA of plasma proteins between 40 and 190 kDa decreased with size for endometrial carcinoma and healthy ovarian tissue, but was independent of molecular weight for the ovarian carcinomas. An effect of charge on distribution volume was only found in healthy ovaries, which had lower hydration and high collagen content, indicating that a condensed interstitium increases the influence of negative charges. A number of earlier suggested biomarker candidates were detected in increased amounts in malignant tissue, e.g., stathmin and spindlin-1, showing that interstitial fluid, even when unfractionated, can be a valuable source for tissue-specific proteins. We demonstrate that the distribution of abundant plasma proteins in the interstitium can be elucidated by mass spectrometry methods and depends markedly on hydration and ECM structure. Our data can be used in modeling of drug uptake, and give indications on ECM components to be targeted to increase the uptake of macromolecular substances.

  4. Immunolocalization of dentin matrix protein-1 in human primary teeth treated with different pulp capping materials.

    Science.gov (United States)

    Lourenço Neto, Natalino; Marques, Nádia C T; Fernandes, Ana Paula; Rodini, Camila O; Sakai, Vivien T; Abdo, Ruy Cesar C; Machado, Maria Aparecida A M; Santos, Carlos F; Oliveira, Thais M

    2016-01-01

    The aim of this study was to evaluate the immunolocalization of dentin matrix protein (DMP)-1 in human primary teeth treated with different pulp capping materials. Twenty-five primary molars were divided into the following groups: formocresol (FC), calcium hydroxide (CH), mineral trioxide aggregate (MTA), corticosteroid/antibiotic solution + CH (O + CH), and Portland cement (PC), and all received conventional pulpotomy treatment. The teeth at the regular exfoliation period were extracted for histological analysis and immunolocalization of DMP-1. Statistical analysis was performed using the χ(2) test (p < 0.05). Histological analysis revealed statistically significant differences in the comparison among the groups through the use of a score system regarding the presence of hard tissue barrier, odontoblastic layer, and internal resorption, but not regarding pulp calcification. Immunohistochemical analysis showed immunostaining for DMP-1 in groups CH, MTA, O + CH, and PC. Internal resorption was observed in the groups FC and CH. MTA and PC showed pulp repair without inflammation and with the presence of hard tissue barrier. DMP-1 immunostaining was higher for MTA and PC, confirming the reparative and bioinductive capacity of these materials.

  5. Amsacrine suppresses matrix metalloproteinase-2 (MMP-2)/MMP-9 expression in human leukemia cells.

    Science.gov (United States)

    Liu, Wen-Hsin; Chen, Ying-Jung; Chien, Jen-Hung; Chang, Long-Sen

    2014-05-01

    This study explores the suppression mechanism of amsacrine (4-(9-Acridinylamino)-N-(methanesulfonyl)-m-anisidine hydrochloride) on matrix metalloproteinase-2 (MMP-2) and MMP-9 expression in human leukemia cells. Amsacrine attenuated cell invasion with decreased MMP-2/MMP-9 protein expression and mRNA levels in U937, Jurkat, HL-60, K562, KU812, and MEG-01 cells. Moreover, amsacrine reduced both MMP-2/MMP-9 promoter luciferase activity and MMP-2/MMP-9 mRNA stability in leukemia cells. Studies on amsacrine-treated U937 cells revealed that amsacrine-elicited ROS generation induced JNK and p38 MAPK activation but reduced the phospho-ERK level. Amsacrine-induced ERK inactivation and p38 MAPK/JNK activation were demonstrated to suppress MMP-2/MMP-9 promoter luciferase activity and promote MMP-2/MMP-9 mRNA decay, respectively. p38 MAPK/JNK activation led to up-regulation of protein phosphatase 2A catalytic subunit α (PP2Acα) in amsacrine-treated U937 cells. Okadaic acid (PP2A inhibitor) treatment increased MMP-2/MMP-9 mRNA stability in amsacrine-treated cells, whereas PP2Acα over-expression increased MMP-2/MMP-9 mRNA decay. Amsacrine-induced MMP-2/MMP-9 down-regulation was also related to PP2Acα up-regulation on Jurkat, HL-60, K562, KU812, and MEG-01 cells. Collectively, our data indicate that amsacrine induces MMP-2/MMP-9 down-regulation via simultaneous suppression of genetic transcription and mRNA stability in human leukemia cells.

  6. Growth and differentiation of human lens epithelial cells in vitro on matrix

    Science.gov (United States)

    Blakely, E. A.; Bjornstad, K. A.; Chang, P. Y.; McNamara, M. P.; Chang, E.; Aragon, G.; Lin, S. P.; Lui, G.; Polansky, J. R.

    2000-01-01

    PURPOSE: To characterize the growth and maturation of nonimmortalized human lens epithelial (HLE) cells grown in vitro. METHODS: HLE cells, established from 18-week prenatal lenses, were maintained on bovine corneal endothelial (BCE) extracellular matrix (ECM) in medium supplemented with basic fibroblast growth factor (FGF-2). The identity, growth, and differentiation of the cultures were characterized by karyotyping, cell morphology, and growth kinetics studies, reverse transcription-polymerase chain reaction (RT-PCR), immunofluorescence, and Western blot analysis. RESULTS: HLE cells had a male, human diploid (2N = 46) karyotype. The population-doubling time of exponentially growing cells was 24 hours. After 15 days in culture, cell morphology changed, and lentoid formation was evident. Reverse transcription-polymerase chain reaction (RT-PCR) indicated expression of alphaA- and betaB2-crystallin, fibroblast growth factor receptor 1 (FGFR1), and major intrinsic protein (MIP26) in exponential growth. Western analyses of protein extracts show positive expression of three immunologically distinct classes of crystallin proteins (alphaA-, alphaB-, and betaB2-crystallin) with time in culture. By Western blot analysis, expression of p57(KIP2), a known marker of terminally differentiated fiber cells, was detectable in exponential cultures, and levels increased after confluence. MIP26 and gamma-crystallin protein expression was detected in confluent cultures, by using immunofluorescence, but not in exponentially growing cells. CONCLUSIONS: HLE cells can be maintained for up to 4 months on ECM derived from BCE cells in medium containing FGF-2. With time in culture, the cells demonstrate morphologic characteristics of, and express protein markers for, lens fiber cell differentiation. This in vitro model will be useful for investigations of radiation-induced cataractogenesis and other studies of lens toxicity.

  7. Cartilage tissue engineering of nasal septal chondrocyte-macroaggregates in human demineralized bone matrix.

    Science.gov (United States)

    Liese, Juliane; Marzahn, Ulrike; El Sayed, Karym; Pruss, Axel; Haisch, Andreas; Stoelzel, Katharina

    2013-06-01

    Tissue Engineering is an important method for generating cartilage tissue with isolated autologous cells and the support of biomaterials. In contrast to various gel-like biomaterials, human demineralized bone matrix (DBM) guarantees some biomechanical stability for an application in biomechanically loaded regions. The present study combined for the first time the method of seeding chondrocyte-macroaggregates in DBM for the purpose of cartilage tissue engineering. After isolating human nasal chondrocytes and creating a three-dimensional macroaggregate arrangement, the DBM was cultivated in vitro with the macroaggregates. The interaction of the cells within the DBM was analyzed with respect to cell differentiation and the inhibitory effects of chondrocyte proliferation. In contrast to chondrocyte-macroaggregates in the cell-DBM constructs, morphologically modified cells expressing type I collagen dominated. The redifferentiation of chondrocytes, characterized by the expression of type II collagen, was only found in low amounts in the cell-DBM constructs. Furthermore, caspase 3, a marker for apoptosis, was detected in the chondrocyte-DBM constructs. In another experimental setting, the vitality of chondrocytes as related to culture time and the amount of DBM was analyzed with the BrdU assay. Higher amounts of DBM tended to result in significantly higher proliferation rates of the cells within the first 48 h. After 96 h, the vitality decreased in a dose-dependent fashion. In conclusion, this study provides the proof of concept of chondrocyte-macroaggregates with DBM as an interesting method for the tissue engineering of cartilage. The as-yet insufficient redifferentiation of the chondrocytes and the sporadic initiation of apoptosis will require further investigations.

  8. Bladder tissue regeneration using acellular bi-layer silk scaffolds in a large animal model of augmentation cystoplasty.

    Science.gov (United States)

    Tu, Duong D; Chung, Yeun Goo; Gil, Eun Seok; Seth, Abhishek; Franck, Debra; Cristofaro, Vivian; Sullivan, Maryrose P; Di Vizio, Dolores; Gomez, Pablo; Adam, Rosalyn M; Kaplan, David L; Estrada, Carlos R; Mauney, Joshua R

    2013-11-01

    Acellular scaffolds derived from Bombyx mori silk fibroin were investigated for their ability to support functional tissue regeneration in a porcine model of augmentation cystoplasty. Two bi-layer matrix configurations were fabricated by solvent-casting/salt leaching either alone (Group 1) or in combination with silk film casting (Group 2) to yield porous foams buttressed by heterogeneous surface pore occlusions or homogenous silk films, respectively. Bladder augmentation was performed with each scaffold group (6 × 6 cm(2)) in juvenile Yorkshire swine for 3 m of implantation. Augmented animals exhibited high rates of survival (Group 1: 5/6, 83%; Group 2: 4/4, 100%) and voluntary voiding over the course of the study period. Urodynamic evaluations demonstrated mean increases in bladder capacity over pre-operative levels (Group 1: 277%; Group 2: 153%) which exceeded nonsurgical control gains (144%) encountered due to animal growth.In addition, animals augmented with both matrix configurations displayed increases in bladder compliance over pre-operative levels(Group 1: 357%; Group 2: 338%) similar to growth-related elevations observed in non-surgical controls (354%) [corrected]. Gross tissue evaluations revealed that both matrix configurations supported extensive de novo tissue formation throughout the entire original implantation site which exhibited ultimate tensile strength similar to nonsurgical counterparts. Histological and immunohistochemical analyses showed that both implant groups promoted comparable extents of smooth muscle regeneration and contractile protein (α-smooth muscle actin and SM22α) expression within defect sites similar to controls. Parallel evaluations demonstrated the formation of a transitional, multi-layered urothelium with prominent cytokeratin, uroplakin, and p63 protein expression in both matrix groups. De novo innervation and vascularization processes were evident in all regenerated tissues indicated by synaptophysin-positive neuronal

  9. Collaborative study on a Guinea pig serological method for the assay of acellular pertussis vaccines.

    Science.gov (United States)

    Winsnes, R; Sesardic, D; Daas, A; Terao, E; Behr-Gross, M-E

    2009-10-01

    An international collaborative study (coded BSP083) was performed under the aegis of the Biological Standardisation Programme supported by the Council of Europe and the European Commission, with the aim of replacing the in vivo challenge assays for potency determination of combined acellular pertussis (aP) vaccines by a refined procedure also allowing reduction of animal use. This study investigates whether the immunogenicity of aP vaccine components could be assayed in a guinea pig (gp) serology model, using the same vaccine immunising doses as for D and T components potency testing, instead of using separate animals as is currently done. The BSP83 project is a follow up of 3 former collaborative studies (coded BSP019, BSP034 and BSP035) on serological methods for the potency testing of tetanus (T) and diphtheria (D) vaccines for human use. The use of gp instead of mice serology has the advantage of providing a larger volume of good quality antiserum for the assay of several vaccine components in the same sample, hence providing the opportunity for animal sparing. The results of Phase I of the study demonstrated that gp serology may be a useful method for the immunogenicity assay of acellular pertussis vaccines. This was confirmed in Phase II of the study, using 7 different combined aP vaccines in an international collaborative study involving 17 laboratories from both public and private sectors. Clear dose-response relationships were observed for different vaccines by ELISA, for antibodies against aP antigens, i.e. pertussis toxin (PT), filamentous haemagglutinin (FHA), fimbrial agglutinogens-2/3 (Fim 2/3) and pertactin (PRN). Intra- and inter-laboratory variations of aP ELISA results were found to be within an acceptable range. For some combined vaccines, however, the range of vaccine dilutions for immunisation confirmed to be optimal for D and T potency testing may not provide optimal dose-response for all aP components. Method adjustments may thus be required

  10. Extracellular Matrix Hydrogel Derived from Human Umbilical Cord as a Scaffold for Neural Tissue Repair and Its Comparison with Extracellular Matrix from Porcine Tissues.

    Science.gov (United States)

    Kočí, Zuzana; Výborný, Karel; Dubišová, Jana; Vacková, Irena; Jäger, Aleš; Lunov, Oleg; Jiráková, Klára; Kubinová, Šárka

    2017-06-01

    Extracellular matrix (ECM) hydrogels prepared by tissue decellularization have been reported as natural injectable materials suitable for neural tissue repair. In this study, we prepared ECM hydrogel derived from human umbilical cord (UC) and evaluated its composition and mechanical and biological properties in comparison with the previously described ECM hydrogels derived from porcine urinary bladder (UB), brain, and spinal cord. The ECM hydrogels did not differ from each other in the concentration of collagen, while the highest content of glycosaminoglycans as well as the shortest gelation time was found for UC-ECM. The elastic modulus was then found to be the highest for UB-ECM. In spite of a different origin, topography, and composition, all ECM hydrogels similarly promoted the migration of human mesenchymal stem cells (MSCs) and differentiation of neural stem cells, as well as axonal outgrowth in vitro. However, only UC-ECM significantly improved proliferation of tissue-specific UC-derived MSCs when compared with the other ECMs. Injection of UC-ECM hydrogels into a photothrombotic cortical ischemic lesion in rats proved its in vivo gelation and infiltration with host macrophages. In summary, this study proposes UC-ECM hydrogel as an easily accessible biomaterial of human origin, which has the potential for neural as well as other soft tissue reconstruction.

  11. Oxidation modifies the structure and function of the extracellular matrix generated by human coronary artery endothelial cells.

    Science.gov (United States)

    Chuang, Christine Y; Degendorfer, Georg; Hammer, Astrid; Whitelock, John M; Malle, Ernst; Davies, Michael J

    2014-04-15

    ECM (extracellular matrix) materials, such as laminin, perlecan, type IV collagen and fibronectin, play a key role in determining the structure of the arterial wall and the properties of cells that interact with the ECM. The aim of the present study was to investigate the effect of peroxynitrous acid, an oxidant generated by activated macrophages, on the structure and function of the ECM laid down by HCAECs (human coronary artery endothelial cells) in vitro and in vivo. We show that exposure of HCAEC-derived native matrix components to peroxynitrous acid (but not decomposed oxidant) at concentrations >1 μM results in a loss of antibody recognition of perlecan, collagen IV, and cell-binding sites on laminin and fibronectin. Loss of recognition was accompanied by decreased HCAEC adhesion. Real-time PCR showed up-regulation of inflammation-associated genes, including MMP7 (matrix metalloproteinase 7) and MMP13, as well as down-regulation of the laminin α2 chain, in HCAECs cultured on peroxynitrous acid-treated matrix compared with native matrix. Immunohistochemical studies provided evidence of co-localization of laminin with 3-nitrotyrosine, a biomarker of peroxynitrous acid damage, in type II-III/IV human atherosclerotic lesions, consistent with matrix damage occurring during disease development in vivo. The results of the present study suggest a mechanism through which peroxynitrous acid modifies endothelial cell-derived native ECM proteins of the arterial basement membrane in atherosclerotic lesions. These changes to ECM and particularly perlecan and laminin may be important in inducing cellular dysfunction and contribute to atherogenesis.

  12. Brainless but Multi-Headed: Decision Making by the Acellular Slime Mould Physarum polycephalum.

    Science.gov (United States)

    Beekman, Madeleine; Latty, Tanya

    2015-11-20

    Because of its peculiar biology and the ease with which it can be cultured, the acellular slime mould Physarum polycephalum has long been a model organism in a range of disciplines. Due to its macroscopic, syncytial nature, it is no surprise that it has been a favourite amongst cell biologists. Its inclusion in the experimental tool kit of behavioural ecologists is much more recent. These recent studies have certainly paid off. They have shown that, for an organism that lacks a brain or central nervous system, P. polycephalum shows rather complex behaviour. For example, it is capable of finding the shortest path through a maze, it can construct networks as efficient as those designed by humans, it can solve computationally difficult puzzles, it makes multi-objective foraging decisions, it balances its nutrient intake and it even behaves irrationally. Are the slime mould's achievements simply "cute", worthy of mentioning in passing but nothing to take too seriously? Or do they hint at the fundamental processes underlying all decision making? We will address this question after reviewing the decision-making abilities of the slime mould. Copyright © 2015 Elsevier Ltd. All rights reserved.

  13. Decellularization of porcine skeletal muscle extracellular matrix for the formulation of a matrix hydrogel: a preliminary study.

    Science.gov (United States)

    Fu, Yuehe; Fan, Xuejiao; Tian, Chunxiang; Luo, Jingcong; Zhang, Yi; Deng, Li; Qin, Tingwu; Lv, Qing

    2016-04-01

    Extracellular matrix (ECM) hydrogels are used as scaffolds to facilitate the repair and reconstruction of tissues. This study aimed to optimize the decellularization process of porcine skeletal muscle ECM and to formulate a matrix hydrogel scaffold. Five multi-step methods (methods A-E) were used to generate acellular ECM from porcine skeletal muscle [rinsing in SDS, trypsin, ethylenediaminetetraacetic acid (EDTA), Triton X-100 and/or sodium deoxycholate at 4-37°C]. The resulting ECM was evaluated using haematoxylin and eosin, 4-6-diamidino-2-phenylindole (DAPI) staining, and DNA quantification. Acellular matrix was dissolved in pepsin and gelled at 37°C. Hydrogel response to temperature was observed in vivo and in vitro. ECM components were assessed by Masson, Sirius red, and alcian blue staining, and total protein content. Acellular porcine skeletal muscle exhibited a uniform translucent white appearance. No intact nuclear residue was detected by haematoxylin and eosin staining, while DAPI staining showed a few nuclei in the matrixes produced by methods B, C, and D. Method A generated a gel that was too thin for gelation. However, the matrix obtained by rinsing in 0.2% trypsin/0.1% EDTA, 0.5% Triton X-100, and 1% Triton X-100/0.2% sodium deoxycholate was nuclei-free and produced a viscous solution that formed a structurally stable white jelly-like hydrogel. The residual DNA content of this solution was 49.37 ± 0.72 ng/mg, significantly less than in fresh skeletal muscle, and decreased to 19.22 ± 0.85 ng/mg after gelation (P collagen and glycosaminoglycan, with a total protein concentration of 64.8 ± 6.9%. An acellular ECM hydrogel from porcine skeletal muscle was efficiently produced.

  14. Human disc cells in monolayer vs 3D culture: cell shape, division and matrix formation

    Directory of Open Access Journals (Sweden)

    Hanley Edward N

    2000-10-01

    Full Text Available Abstract Background The relationship between cell shape, proliferation, and extracellular matrix (ECM production, important aspects of cell behavior, is examined in a little-studied cell type, the human annulus cell from the intervertebral disc, during monolayer vs three-dimensional (3D culture. Results Three experimental studies showed that cells respond specifically to culture microenvironments by changes in cell shape, mitosis and ECM production: 1 Cell passages showed extensive immunohistochemical evidence of Type I and II collagens only in 3D culture. Chondroitin sulfate and keratan sulfate were abundant in both monolayer and 3D cultures. 2 Cells showed significantly greater proliferation in monolayer in the presence of platelet-derived growth factor compared to cells in 3D. 3 Cells on Matrigel™-coated monolayer substrates became rounded and formed nodular colonies, a finding absent during monolayer growth. Conclusions The cell's in vivo interactions with the ECM can regulate shape, gene expression and other cell functions. The shape of the annulus cell changes markedly during life: the young, healthy disc contains spindle shaped cells and abundant collagen. With aging and degeneration, many cells assume a strikingly different appearance, become rounded and are surrounded by unusual accumulations of ECM products. In vitro manipulation of disc cells provides an experimental window for testing how disc cells from given individuals respond when they are grown in environments which direct cells to have either spindle- or rounded-shapes. In vitro assessment of the response of such cells to platelet-derived growth factor and to Matrigel™ showed a continued influence of cell shape even in the presence of a growth factor stimulus. These findings contribute new information to the important issue of the influence of cell shape on cell behavior.

  15. Suspension Culture Alters Insulin Secretion in Induced Human Umbilical Cord Matrix-Derived Mesenchymal Cells

    Directory of Open Access Journals (Sweden)

    Fatemeh Seyedi

    2016-04-01

    Full Text Available Objective: Worldwide, diabetes mellitus (DM is an ever-increasing metabolic disorder. A promising approach to the treatment of DM is the implantation of insulin producing cells (IPC that have been derived from various stem cells. Culture conditions play a pivotal role in the quality and quantity of the differentiated cells. In this experimental study, we have applied various culture conditions to differentiate human umbilical cord matrix-derived mesenchymal cells (hUCMs into IPCs and measured insulin production. Materials and Methods: In this experimental study, we exposed hUCMs cells to pancreatic medium and differentiated them into IPCs in monolayer and suspension cultures. Pancreatic medium consisted of serum-free Dulbecco’s modified eagle’s medium Nutrient mixture F12 (DMEM/F12 medium with 17.5 mM glucose supplemented by 10 mM nicotinamide, 10 nM exendin-4, 10 nM pentagastrin, 100 pM hepatocyte growth factor, and B-27 serum-free supplement. After differentiation, insulin content was analyzed by gene expression, immunocytochemistry (IHC and the chemiluminesence immunoassay (CLIA. Results: Reverse transcription-polymerase chain reaction (RT-PCR showed efficient expressions of NKX2.2, PDX1 and INSULIN genes in both groups. IHC analysis showed higher expression of insulin protein in the hanging drop group, and CLIA revealed a significant higher insulin production in hanging drops compared with the monolayer group following the glucose challenge test. Conclusion: We showed by this novel, simple technique that the suspension culture played an important role in differentiation of hUCMs into IPC. This culture was more efficient than the conventional culture method commonly used in IPC differentiation and cultivation.

  16. Marked induction of matrix metalloproteinase-10 by respiratory syncytial virus infection in human nasal epithelial cells.

    Science.gov (United States)

    Hirakawa, Satoshi; Kojima, Takashi; Obata, Kazufumi; Okabayashi, Tamaki; Yokota, Shin-Ichi; Nomura, Kazuaki; Obonai, Toshimasa; Fuchimoto, Jun; Himi, Tetsuo; Tsutsumi, Hiroyuki; Sawada, Norimasa

    2013-12-01

    Respiratory syncytial virus (RSV) is an important pathogen of bronchiolitis, asthma, and severe lower respiratory tract disease in infants and young children. Matrix metalloproteinases (MMPs) play key roles in viral infection, inflammation and remodeling of the airway. However, the roles and regulation of MMPs in human nasal epithelial cells (HNECs) after RSV infection remain unclear. To investigate the regulation of MMP induced after RSV infection in HNECs, an RSV-infected model of HNECs in vitro was used. It was found that mRNA of MMP-10 was markedly increased in HNECs after RSV infection, together with induction of mRNAs of MMP-1, -7, -9, and -19. The amount of MMP-10 released from HNECs was also increased in a time-dependent manner after RSV infection as was that of chemokine RANTES. The upregulation of MMP-10 in HNECs after RSV infection was prevented by inhibitors of NF-κB and pan-PKC with inhibition of RSV replication, whereas it was prevented by inhibitors of JAK/STAT, MAPK, and EGF receptors without inhibition of RSV replication. In lung tissue of an infant with severe RSV infection in which a few RSV antibody-positive macrophages were observed, MMP-10 was expressed at the apical side of the bronchial epithelial cells and alveolar epithelial cells. In conclusion, MMP-10 induced by RSV infection in HNECs is regulated via distinct signal transduction pathways with or without relation to RSV replication. MMP-10 may play an important role in the pathogenesis of RSV diseases and it has the potential to be a novel marker and therapeutic target for RSV infection.

  17. CD14{sup +} monocytes promote the immunosuppressive effect of human umbilical cord matrix stem cells

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Ding, E-mail: qqhewd@gmail.com [The State Key Laboratory of Experimental Hematology, Institute of Hematology and Hospital of Blood Diseases, Chinese Academy of Medical Sciences and Peking Union of Medical College, 288 Nanjing Road, Tianjin 300020 (China); TEDA Life and Technology Research Center, Institute of Hematology, Chinese Academy of Medical Sciences, TEDA, Tianjin (China); Chen, Ke, E-mail: chenke_59@hotmail.com [The State Key Laboratory of Experimental Hematology, Institute of Hematology and Hospital of Blood Diseases, Chinese Academy of Medical Sciences and Peking Union of Medical College, 288 Nanjing Road, Tianjin 300020 (China); TEDA Life and Technology Research Center, Institute of Hematology, Chinese Academy of Medical Sciences, TEDA, Tianjin (China); Du, Wei Ting, E-mail: duwtpumc@yahoo.com.cn [The State Key Laboratory of Experimental Hematology, Institute of Hematology and Hospital of Blood Diseases, Chinese Academy of Medical Sciences and Peking Union of Medical College, 288 Nanjing Road, Tianjin 300020 (China); Han, Zhi-Bo, E-mail: zhibohan@hotmail.com [The State Key Laboratory of Experimental Hematology, Institute of Hematology and Hospital of Blood Diseases, Chinese Academy of Medical Sciences and Peking Union of Medical College, 288 Nanjing Road, Tianjin 300020 (China); TEDA Life and Technology Research Center, Institute of Hematology, Chinese Academy of Medical Sciences, TEDA, Tianjin (China); Ren, He, E-mail: knifesharp2000@hotmail.com [National Engineering Research Center of Cell Products, AmCellGene Co. Ltd, TEDA, Tianjin (China); Chi, Ying, E-mail: caizhuying@hotmail.com [The State Key Laboratory of Experimental Hematology, Institute of Hematology and Hospital of Blood Diseases, Chinese Academy of Medical Sciences and Peking Union of Medical College, 288 Nanjing Road, Tianjin 300020 (China); TEDA Life and Technology Research Center, Institute of Hematology, Chinese Academy of Medical Sciences, TEDA, Tianjin (China); and others

    2010-09-10

    Here, the effect of CD14{sup +} monocytes on human umbilical cord matrix stem cell (hUC-MSC)-mediated immunosuppression was studied in vitro. hUC-MSCs exerted a potent inhibitory effect on the proliferation and interferon-{gamma} (IFN-{gamma}) secretion capacities of CD4{sup +} and CD8{sup +} T cells in response to anti-CD3/CD28 stimulation. Transwell co-culture system revealed that the suppressive effect was primarily mediated by soluble factors. Addition of prostaglandin synthesis inhibitors (indomethacin or NS-398) almost completely abrogated the immunosuppression activity of hUC-MSCs, identifying prostaglandin E{sub 2} (PGE{sub 2}) as an important soluble mediator. CD14{sup +} monocytes were found to be able to enhance significantly the immunosuppressive effect of hUC-MSCs in a dose-dependent fashion. Moreover, the inflammatory cytokine IL-1{beta}, either exogenously added or produced by CD14{sup +} monocytes in culture, could trigger expression of high levels of PGE{sub 2} by hUC-MSCs, whereas inclusion of the IL-1 receptor antagonist (IL-1RA) in the culture down-regulated not only PGE{sub 2} expression, but also reversed the promotional effect of CD14{sup +} monocytes and partially restored CD4{sup +} and CD8{sup +} T cell proliferation and IFN-{gamma} secretion. Our data demonstrate an important role of monocytes in the hUC-MSC-induced immunomodulation, which may have important implications in future efforts to explore the clinical potentials of hUC-MSCs.

  18. A retrospective analysis of a human cellular repair matrix for the treatment of chronic wounds.

    Science.gov (United States)

    Regulski, Matthew; Jacobstein, Douglas A; Petranto, Russell D; Migliori, Vincent J; Nair, Girish; Pfeiffer, Darelle

    2013-12-01

    Despite the introduction of advanced wound care modalities over the last 15 years, chronic wounds are an increasing problem. Few single options are available for clinicians to treat recalcitrant wounds such as diabetic foot ulcers (DFUs) and venous leg ulcers (VLUs). A retrospective, single-center study was conducted at an outpatient wound care center to evaluate the clinical effect of a human cellular repair matrix (h-CRM) on chronic wounds that had failed to heal. Data from all patients who had received this treatment modality during a period of 2 years were abstracted. Standard care included weekly visits, regular debridement, offloading DFUs, compression for VLUs, and h-CRM for wounds >4 weeks duration. A total of 66 patients (30 male, 36 female, mean age 71.1 [± 8.8] years) received h-CRM treatment for 67 wounds (34 VLUs, 27 DFUs, and six other chronic wounds). The average wound size at baseline was 6.65 (± 9.68) cm2, and the average wound duration before h-CRM treatment was 38 (±70.8) weeks. Fifty (50) patients (74.6%) had failed to heal using other advanced therapies. After 12 weeks of care, 51 of the 67 wounds (76.1%) were healed: 23 of 34 (67.6%) VLUs and 23 of 27 (85.2%) DFUs. Average time to closure in these wounds was 5.8 (±2.5) weeks. No significant differences were found between proportions of VLUs and DFUs healed. No adverse events or recurrences occurred during an average follow-up time of 20.4 months (range 11 to 32 months). Overall, patients received an average of 3.8 applications of h-CRM, and 3.2 applications were used among patients that healed. The study results suggest h-CRM may benefit patients with chronic wounds. Prospective, randomized clinical studies are warranted.

  19. Levels of matrix metalloproteinase-7 and osteopontin in human gingival crevicular fluid during initial tooth movement

    Directory of Open Access Journals (Sweden)

    Dhaval Oswal

    2015-01-01

    Full Text Available Purpose: During orthodontic treatment, the early response of periodontal tissues to mechanical stress involves several metabolic changes that allow tooth movement. The purpose of this investigation was to evaluate osteopontin (OPN and matrix metalloproteinase (MMP-7 in the gingival crevicular fluid (GCF of human teeth exposed to orthodontic force. Materials and Methods: GCF samples were obtained from 15 healthy orthodontic patients (age, 12-22 years. In each patient, the left maxillary canine having the fixed orthodontic appliance was used as the test tooth, and its antagonist, with no appliance, was the control. Orthodontic force, 75 g was applied using a 16 × 22 beta titanium closing loop. The GCF sampling on the disto-buccal aspects of experimental and control tooth was performed at specific time interval with sterilized absorbent paper point. Processing was carried out with enzyme-linked immunosorbent assay to detect OPN and MMP-7 levels. Results: The peak level of OPN was seen after 1 h application of orthodontic force which was 1280.36 pg/ml ± 185.02. The peak level of MMP-7 was seen at 0 h which was 598.3 pg/ml ± 107.5. The levels of OPN after 1 h increased to 1280.36 pg/ml ± 185.02, and they decreased at 24 h to 1012.86 pg/ml ± 168.47 (P = 0.001. The levels of MMP-7 after 1 h decreased to 478 pg/ml ± 99.7 which increased at 24 h to 526.9 pg/ml ± 99.2. Conclusions: Orthodontic forces affect both OPN and MMP-7 protein levels on the compression side in a time-dependent fashion.

  20. Extracellular matrix sub-types and mechanical stretch impact human cardiac fibroblast responses to transforming growth factor beta.

    Science.gov (United States)

    Watson, Chris J; Phelan, Dermot; Collier, Patrick; Horgan, Stephen; Glezeva, Nadia; Cooke, Gordon; Xu, Maojia; Ledwidge, Mark; McDonald, Kenneth; Baugh, John A

    2014-06-01

    Understanding the impact of extracellular matrix sub-types and mechanical stretch on cardiac fibroblast activity is required to help unravel the pathophysiology of myocardial fibrotic diseases. Therefore, the purpose of this study was to investigate pro-fibrotic responses of primary human cardiac fibroblast cells exposed to different extracellular matrix components, including collagen sub-types I, III, IV, VI and laminin. The impact of mechanical cyclical stretch and treatment with transforming growth factor beta 1 (TGFβ1) on collagen 1, collagen 3 and alpha smooth muscle actin mRNA expression on different matrices was assessed using quantitative real-time PCR. Our results revealed that all of the matrices studied not only affected the expression of pro-fibrotic genes in primary human cardiac fibroblast cells at rest but also affected their response to TGFβ1. In addition, differential cellular responses to mechanical cyclical stretch were observed depending on the type of matrix the cells were adhered to. These findings may give insight into the impact of selective pathological deposition of extracellular matrix proteins within different disease states and how these could impact the fibrotic environment.

  1. Potential sites for the perception of gravity in the acellular slime mold Physarum polycephalum

    Science.gov (United States)

    Block, I.; Briegleb, W.

    Recently a gravisensitivity of the acellular slime mold Physarum polycephalum, which possesses no specialized gravireceptor, could be established by conducting experiments under simulated and under real near weightlessness. In these experiments macroplasmodia showed a modulation of their contraction rhythm followed by regulation phenomena. Until now the perception mechanism for the gravistimulus is unknown, but several findings indicate the involvement of mitochondria: A) During the impediment of respiration the Og-reaction is inhibited and the regulation is reduced. B) The response to a light stimulus and the following regulation phenomena strongly resemble the behavior during exposure to Og, the only difference is that the two reactions are directed into opposite directions. In the blue-light reaction a flavin of the mitochondrial matrix seems to be involved in the light perception. C) The contraction rhythm as well as its modulations are coupled to rhythmic changes in the levels of ATP and calcium ions, involving the mitochondria as sites of energy production and of Ca++-storage. - So the mitochondria could be the site of the regulation and they possibly are the receptor sites for the light and gravity stimuli. - Also the observation of a morphologic polarity of the slime mold's plasmodial strands has to be considered: Cross-sections reveal that the ectoplasmic wall surrounding the streaming endoplasm is much thinner on the physically lower side than on the upper side of the strand - this applies to strands lying on or hanging on a horizontal surface. So, in addition to the mitochondria, also the morphologic polarity may be involved in the perception mechanism of the observed gravisensitivity and of the recently established geotaxis. - The potential role of the nuclei and of the contractile elements in the perception of gravity is also discussed.

  2. Doxycycline inhibits neutrophil (PMN)-type matrix metalloproteinases in human adult periodontitis gingiva.

    Science.gov (United States)

    Golub, L M; Sorsa, T; Lee, H M; Ciancio, S; Sorbi, D; Ramamurthy, N S; Gruber, B; Salo, T; Konttinen, Y T

    1995-02-01

    We previously reported that low-dose doxycycline (DOXY) therapy reduces host-derived collagenase activity in gingival tissue of adult periodontitis (AP) patients. However, it was not clear whether this in vivo effect was direct or indirect. In the present study, inflamed human gingival tissue, obtained from AP patients during periodontal surgery, was extracted and the extracts partially purified by (NH4)2SO4 precipitation. The extracts were then analyzed for collagenase activity using SDS-PAGE/fluorography/laser densitometry, and for gelatinase activity using type I gelatin zymography as well as a new quantitative assay using biotinylated type I gelatin as substrate. DOXY was added to the incubation mixture at a final concentration of 0-1000 microM. The concentration of DOXY required to inhibit 50% of the gingival tissue collagenase (IC50) was found to be 16-18 microM in the presence or absence of 1.2 mM APMA (an optimal organomercurial activator of latent procollagenases); this IC50 for DOXY was similar to that exhibited for collagenase or matrix metalloproteinase (MMP)-8 from polymorphonuclear leukocytes (PMNs) and from gingival crevicular fluid (GCF) of AP patients. Of interest, Porphyromonas gingivalis collagenase was also inhibited by similar DOXY levels (IC50 = 15 microM), however the collagenase activity observed in the gingival tissue extracts was found to be of mammalian not bacterial origin based on the production of the specific alpha A (3/4) and alpha B (1/4) collagen degradation fragments. In contrast, the inhibition of collagenase purified from culture media of human gingival fibroblasts (MMP-1) required much greater DOXY levels (IC50 = 280 microM). The predominant molecular forms of gelatinolytic activity presented in the AP patients gingival tissue extracts were found to closely correspond to the 92 kD PMN-type gelatinase (MMP-9) although small quantities of 72 kD fibroblast-type gelatinase (MMP-2), and some other low molecular weight gelatinases

  3. Influence of covering of auto-crosslinked sodium hyaluronate gel in combination with xenogenic acellular dermal matrix on healing of full-thickness skin defect wound in pig%自交联透明质酸钠凝胶联合异种脱细胞真皮基质覆盖对猪全层皮肤缺损创面愈合的影响

    Institute of Scientific and Technical Information of China (English)

    邱宇轩; 张国安; 万江波; 赵筱卓

    2016-01-01

    .0±3.8)%,明显低于其余3组(P值均小于0.05). 结论 自交联透明质酸钠凝胶联合羊ADM作为猪全层皮肤缺损创面微粒皮移植覆盖物延长了羊ADM在创面上的存留时间,促进了创面愈合.%Objective To explore the influence of covering of auto-crosslinked sodium hyaluronate gel in combination with xenogenic acellular dermal matrix (ADM) on healing of full-thickness skin defect wound in pig.Methods Totally four 10 cm × 10 cm full-thickness skin defect wounds were reproduced symmetrically on both sides of spine on the back of each one of the six Chinese experimental minipigs.After autologous microskin grafting,the 4 wounds in each pig were divided into 4 groups according to the random number table,with 6 wounds in each group.Wounds in allogenic skin group (AS) were covered by fullthickness skin from one (not the recipient) of the 6 pigs;wounds in xenogenic skin group (XS) were covered by full-thickness skin of sheep;wounds in xenogenic ADM group (XA) were covered by ADM of sheep;wounds in combination group (C) were covered by ADM of sheep combined with auto-crosslinked sodium hyaluronate gel.The wounds were bound up with pressure,and the dressing was changed once every 7 days.On post surgery day (PSD) 7,14,21,28,35,and 42 when changing dressing,the condition of wounds and the exfoliation of the covering on microskin were observed,and the complete exfoliation time of the covering was recorded.On PSD 28,35,and 42,the wound healing rate was calculated.Data were processed with one-way analysis of variance and SNK test.Results (1) On PSD 7,no fluid appeared under the covering of wounds in groups AS and C,while plenty of fluid appeared under the covering of wounds in groups XS and XA.From PSD 14 to 35,most of the full-thickness skin of pig in group AS did not exfoliate.All the full-thickness skin of sheep in group XS exfoliated,leaving a lot of crusts on the surface of the wounds on PSD 14.Most of the ADM of sheep in group XA separated

  4. Multispectral Optoacoustic Tomography of Matrix Metalloproteinase Activity in Vulnerable Human Carotid Plaques

    NARCIS (Netherlands)

    Razansky, Daniel; Harlaar, Niels J.; Hillebrands, Jan Luuk; Taruttis, Adrian; Herzog, Eva; Zeebregts, Clark J.; van Dam, Gooitzen M.; Ntziachristos, Vasilis

    Elevated expression of cathepsins, integrins and matrix metalloproteinases (MMPs) is typically associated with atherosclerotic plaque instability. While fluorescent tagging of such molecules has been amply demonstrated, no imaging method was so far shown capable of resolving these

  5. Application of nanodiamonds in human body fluid analysis by matrix-assisted laser desorption/ionization mass spectrometry

    Institute of Scientific and Technical Information of China (English)

    Xianglei Kong

    2008-01-01

    Direct mass spectrometric analysis of complex biological samples is very important and challenging. In this paper, nanodiamonds have been successfully used in matrix-assisted laser desorption/ionization mass spectrometric analysis of human serum and urine. As a practical tool and platform, it can be widely used in the field of humoral proteomics, and it plays a very promising role in clinical diagnosis, including identification of novel disease-associated biomarkers.

  6. Direct delayed breast reconstruction with TAP flap, implant and acellular dermal matrix (TAPIA)

    DEFF Research Database (Denmark)

    Børsen-Koch, Mikkel; Gunnarsson, Gudjon L; Udesen, Ann;

    2015-01-01

    BACKGROUND: The latissimus dorsi (LD) flap is considered one of the working horses within the field of breast reconstruction and it offers several advantages. However, donor-site morbidity may pose a problem. This article describes a new and modified technique for delayed breast reconstruction co...... there is a learning curve, this simple modified technique does not demand any perforator or other vessel dissection. Any trained plastic surgeon should be able to adopt the technique into the growing armamentarium of breast reconstruction possibilities....

  7. Acellular Dermal Matrix in Reconstructive Breast Surgery: Survey of Current Practice among Plastic Surgeons

    Directory of Open Access Journals (Sweden)

    Ahmed M. S. Ibrahim, MD

    2015-04-01

    Conclusions: Plastic surgeons currently use ADM in breast reconstruction for both immediate and staged procedures. Of those responding, a majority of plastic surgeons will incorporate drains and use postoperative antibiotics for more than 48 hours.

  8. An acellular dermal matrix allograft (Alloderm®) for increasing keratinized attached gingiva: A case series

    Science.gov (United States)

    Agarwal, Chitra; Kumar, Baron Tarun; Mehta, Dhoom Singh

    2015-01-01

    Context: Adequate amount of keratinized gingiva is necessary to keep gingiva healthy and free of inflammation. Autografts have been used for years with great success to increase the width of attached gingiva. Autografts, however, have the disadvantage of increasing postoperative morbidity and improper color match with the adjacent tissues. Alloderm® allograft has been introduced as an alternative to autografts to overcome these disadvantages. Aim: In this study, the efficacy of alloderm® in increasing the width of attached gingiva and the stability of gained attached gingiva was evaluated clinically. Materials and Methods: Five patients with sites showing inadequate width of attached gingiva (≤1 mm) were enrolled for the study. The width of keratinized gingiva and other clinical parameters were recorded at baseline and 9th month postoperatively. Result: In all cases, there is the average increase of about 2.5 mm of attached gingiva and was maintained for 9-month. Percentage shrinkage of the graft is about 75% at the end of 3rd month in all cases. Excellent colors match with adjacent tissue has been obtained. Conclusion: The study signifies that Alloderm® results in an adequate increase in the amount of attached gingiva and therefore can be used successfully in place of autografts. PMID:26015676

  9. An acellular dermal matrix allograft (Alloderm ® for increasing keratinized attached gingiva: A case series

    Directory of Open Access Journals (Sweden)

    Chitra Agarwal

    2015-01-01

    Full Text Available Context: Adequate amount of keratinized gingiva is necessary to keep gingiva healthy and free of infl ammation. Autografts have been used for years with great success to increase the width of attached gingiva. Autografts, however, have the disadvantage of increasing postoperative morbidity and improper color match with the adjacent tissues. Alloderm ® allograft has been introduced as an alternative to autografts to overcome these disadvantages. Aim: In this study, the efficacy of alloderm ® in increasing the width of attached gingiva and the stability of gained attached gingiva was evaluated clinically. Materials and Methods: Five patients with sites showing inadequate width of attached gingiva (≤1 mm were enrolled for the study. The width of keratinized gingiva and other clinical parameters were recorded at baseline and 9th month postoperatively. Result: In all cases, there is the average increase of about 2.5 mm of attached gingiva and was maintained for 9-month. Percentage shrinkage of the graft is about 75% at the end of 3 rd month in all cases. Excellent colors match with adjacent tissue has been obtained. Conclusion: The study signifi es that Alloderm ® results in an adequate increase in the amount of attached gingiva and therefore can be used successfully in place of autografts.

  10. Tissue engineering of the small intestine by acellular collagen sponge scaffold grafting.

    Science.gov (United States)

    Hori, Y; Nakamura, T; Matsumoto, K; Kurokawa, Y; Satomi, S; Shimizu, Y

    2001-01-01

    Tissue engineering of the small intestine will prove a great benefit to patients suffering from short bowel disease. However cell seeding in tissue engineering, such as fetal cell use, is accompanied by problems of ethical issues, rejection, and short supply. To overcome these problems, we carried out an experimental study on tissue engineering of the small intestine by acellular collagen sponge scaffold grafting. We resected the 5 cm long jejunum from beagle dogs and reconstructed it by acellular collagen sponge grafting with a silicon tube stent. The graft was covered with the omentum. At 1 month after operation, the silicon stent was removed endoscopically. Animals were sacrificed 1 and 4 months after operation, and were examined microscopically. Neo-intestinal regeneration was observed and the intestinal mucosa covered the luminal side of the regenerated intestine across the anastomosis. Thus, the small intestine was regenerated by tissue engineering technology using an acellular collagen sponge scaffold.

  11. Acellular comet assay: a tool for assessing variables influencing the alkaline comet assay.

    Science.gov (United States)

    Kennedy, Erin K; McNamee, James P; Prud'homme Lalonde, Louise; Jones, Trevor; Wilkinson, Diana

    2012-01-01

    In this study, an acellular modification to the alkaline comet assay to further evaluate key variables within the assay that may influence the outcome of genotoxicity studies is described. This acellular comet assay can detect differences of 0.2 Gy of (60)Co gamma-ray radiation between 0 and 1 Gy and differences of 1 Gy between 0 and 8 Gy; thus, this assay is applicable for a wide range of DNA damage levels. It is also shown that DNA damage from different radiation energies was not significantly different from (60)Co gamma-ray. This assay displayed a statistical increase in DNA damage due to uncontrolled exposure to natural light; however, the slope of the dose-response curve for light-exposed samples was similar to that for samples protected from light. A comparison of the alkaline comet assay with the acellular comet assay allowed for the intrinsic repair capacity of the alkaline comet assay to be quantified.

  12. Pertactin deficient Bordetella pertussis present a better fitness in mice immunized with an acellular pertussis vaccine.

    Science.gov (United States)

    Hegerle, N; Dore, G; Guiso, N

    2014-11-20

    Bordetella pertussis is the etiologic agent of whooping cough and has been the target of vaccination for over fifty years. The latest strategies include the use of acellular pertussis vaccines that induce specific immunity against few virulence factors amongst which pertactin is included in three and five component acellular pertussis vaccines. Recently, it has been reported that B. pertussis clinical isolates loose the production of this adhesin in regions reaching high vaccine coverage with vaccines targeting this virulence factor. We here demonstrate that isolates not producing pertactin are capable of sustaining longer infection as compared to pertactin producing isolates in an in vivo model of acellular pertussis immunization. Loosing pertactin production might thus provide a selective advantage to these isolates in this background, which could account for the upraise in prevalence of these pertactin deficient isolates in the population.

  13. 小鼠胚胎干细胞条件培养液培养的人角膜内皮细胞在脱细胞猪角膜基质上单层细胞片的构建%Formation of cell sheet on acellular porcine corneal stroma with human corneal endothelial cells cocultured by mouse embryonic stem cell conditioned medium

    Institute of Scientific and Technical Information of China (English)

    鹿晓燕; 王智崇

    2016-01-01

    Background Corneal transplantation faces a great challenge because of the shortage of corneal donors and difficulty of human corneal endothelial cells (HCECs) regeneration in vitro.So the study on tissue engineering cornea is still a main topic.Previous research showed that mouse embryonic stem cell conditioned medium (ESC-CM) improved the proliferative capacity of HCECs in vitro,and acellular porcine corneal stroma (APCS) was a good saffold material.However,whether HECEs cultured by mouse ESC-CM can form cell sheet in vitro were rarely studied.Objective This study was to investigate the potential that HCECs cultured by mouse ESC-CM form a monolayer cell sheet.Methods The supernatant of ESC-CM was collected after mouse ES-E14 cells were cultured,and the cultured medium was centrifuged and mixed with 75% human corneal endothelium medium (CEM)at a proportion of 1 ∶ 3 to prepare the 25% ESC-CM system.Primary cultures of HCECs were established from explants of corneal limbal with Descemet's membrane,and the cells were identified by using reverse-transcription PCR to determine the expressions of collagen Ⅷ (Col Ⅷ) mRNA and neuron-specific enolase (NSE) mRNA in the cells.APCS was prepared by decellularization with phospholipase A2 and bicarbonate solution,and the second generation of HCECs were inoculated on the sterilized APCS at a 800/mm2 density.The morphology of the cells was observed by hematoxylin-eosin staining under the phase-contrast microscope.The expressions of zona occludens protein-1 (ZO-1)and Na+-K+-ATPase in the cell sheet were detected by immunofluorescence staining.Results The second generation of HCECs cultured with 25% ESC-CM in vitro showed the hexagon in shape with positive expressions for Col Ⅷ mRNA and NSE mRNA.Decellularization APCS was transparent,and no corneal cells were seen,the structures of corneal collagenous fibres were regular.HCECs attached closely to APCS and formed monolayer sheet 7 days after culture on the APCS with the

  14. Extracellular matrix elasticity modulates TGF-β-induced p38 activation and myofibroblast transdifferentiation in human tenon fibroblasts.

    Science.gov (United States)

    Meyer-ter-Vehn, Tobias; Han, Hong; Grehn, Franz; Schlunck, Günther

    2011-11-25

    Extracellular matrix and the cytokine TGF-β influence scar formation in an interdependent fashion. In this study, the impact of extracellular matrix elasticity on TGF-β-induced signal transduction and myofibroblast transdifferentiation was examined. Primary human tenon fibroblasts were seeded on collagen-coated glass coverslips (rigid environment) or collagen or polyacrylamide gels (elastic environment) of different compliance and stimulated with TGF-β. Myofibroblast transdifferentiation was assessed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Western blot analysis for the marker gene α-smooth muscle actin (SMA), and SMA incorporation into stress fibers was determined by confocal immunofluorescence microscopy. CTGF transcription was assessed by RT-qPCR. Signaling pathways were examined by Western blot using phosphospecific antibodies and by immunofluorescence microscopy. TGF-β-dependent myofibroblast transdifferentiation was enhanced in a stiff environment. Increasing matrix elasticity attenuated TGF-β-induced myofibroblast transdifferentiation and the associated CTGF expression. TGF-β-induced p38 activation was reduced on elastic substrates. The results suggest that matrix elasticity influences TGF-β-dependent activation of p38 signaling and subsequent myofibroblast transdifferentiation. Biomechanical cues represent an important determinant of scarring processes. Therefore, cellular signals elicited by mechanotransduction deserve consideration in the design of novel antifibrotic strategies.

  15. MiR-15a-5p regulates viability and matrix degradation of human osteoarthritis chondrocytes via targeting VEGFA.

    Science.gov (United States)

    Chen, Hongwei; Tian, Yun

    2017-01-16

    Previous studies demonstrated that miR-15a-5p was probably associated with human hepatocellular carcinoma, while the function of miR-15a-5p in OA (Osteoarthritis) still remains unknown. Here, we uncovered the potential role of miR-15a-5p on OA pathogenesis and confirmed its predicted target VEGFA (Vascular Endothelial Growth Factor A). Measured by RT-PCR, miR-15a-5p expression increased remarkably while VEGFA expression was significantly decreased in OA chondrocytes compared with normal conditions. According to Luciferase activity assay, miR-15a-5p directly targeted the 3'-UTR of VEGFA to inhibit its expression. Functional analysis including CCK-8 assay and flow cytometry revealed that overexpression of VEGFA or inhibition of miR-15a-5p promoted cell proliferation, suppressed cell apoptosis and reduced matrix degradation in OA chondrocytes. Moreover, rescue assays carried out with both expression of VEGFA and miR-15a-5p demonstrated that miR-15a-5p contributes to cell apoptosis and matrix degradation via inhibiting VEGFA. We further provided evidence that multiple proteins related to matrix synthesis were regulated by miR-15a-5p and VEGFA using Western blot and ELISA assays. Taken together, our findings elucidated an underlying mechanism by which miR-15a-5p regulates viability and matrix degradation of OA and indicated a new target for OA diagnosis and therapy.

  16. Acellular biological tissues containing inherent glycosaminoglycans for loading basic fibroblast growth factor promote angiogenesis and tissue regeneration.

    Science.gov (United States)

    Lai, Po-Hong; Chang, Yen; Chen, Sung-Ching; Wang, Chung-Chi; Liang, Huang-Chien; Chang, Wei-Chun; Sung, Hsing-Wen

    2006-09-01

    It was found in our previous study that acellular tissues derived from bovine pericardia consist primarily of insoluble collagen, elastin, and tightly bound glycosaminoglycans (GAGs). It is speculated that the inherent GAGs in acellular tissues may serve as a reservoir for loading basic fibroblast growth factor (bFGF) and promote angiogenesis and tissue regeneration. This study was therefore designed to investigate effects of the content of GAGs in acellular bovine pericardia on the binding of bFGF and its release profile in vitro while its stimulation in angiogenesis and tissue regeneration in vivo were evaluated subcutaneously in a rat model. To control the content of GAGs, acellular tissues were treated additionally with hyaluronidase for 1 (Hase-D1), 3 (Hase-D3), or 5 days (Hase-D5). The in vitro results indicated that a higher content of GAGs in the acellular tissue resulted in an increase in bFGF binding and in a more gradual and sustained release of the growth factor. The in vivo results obtained at 1 week postoperatively showed that the density and the depth of neo-vessels infiltrated into the acellular tissue loaded with bFGF (acellular/bFGF) were significantly greater than the other test samples. At 1 month postoperatively, vascularized neo-connective tissues were found to fill the pores within each test sample, particularly for the acellular/bFGF tissue. These results suggested that the sustained release of bFGF from the acellular/ bFGF tissue continued to be effective in enhancing angiogenesis and generation of new tissues. In conclusion, the inherent GAGs present in acellular tissues may be used for binding and sustained release of bFGF to enhance angiogenesis and tissue regeneration.

  17. Peeping into human renal calcium oxalate stone matrix: characterization of novel proteins involved in the intricate mechanism of urolithiasis.

    Directory of Open Access Journals (Sweden)

    Kanu Priya Aggarwal

    Full Text Available BACKGROUND: The increasing number of patients suffering from urolithiasis represents one of the major challenges which nephrologists face worldwide today. For enhancing therapeutic outcomes of this disease, the pathogenic basis for the formation of renal stones is the need of hour. Proteins are found as major component in human renal stone matrix and are considered to have a potential role in crystal-membrane interaction, crystal growth and stone formation but their role in urolithiasis still remains obscure. METHODS: Proteins were isolated from the matrix of human CaOx containing kidney stones. Proteins having MW>3 kDa were subjected to anion exchange chromatography followed by molecular-sieve chromatography. The effect of these purified proteins was tested against CaOx nucleation and growth and on oxalate injured Madin-Darby Canine Kidney (MDCK renal epithelial cells for their activity. Proteins were identified by Matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF MS followed by database search with MASCOT server. In silico molecular interaction studies with CaOx crystals were also investigated. RESULTS: Five proteins were identified from the matrix of calcium oxalate kidney stones by MALDI-TOF MS followed by database search with MASCOT server with the competence to control the stone formation process. Out of which two proteins were promoters, two were inhibitors and one protein had a dual activity of both inhibition and promotion towards CaOx nucleation and growth. Further molecular modelling calculations revealed the mode of interaction of these proteins with CaOx at the molecular level. CONCLUSIONS: We identified and characterized Ethanolamine-phosphate cytidylyltransferase, Ras GTPase-activating-like protein, UDP-glucose:glycoprotein glucosyltransferase 2, RIMS-binding protein 3A, Macrophage-capping protein as novel proteins from the matrix of human calcium oxalate stone which play a critical role in kidney stone

  18. Comparative adherence of Candida albicans and Candida dubliniensis to human buccal epithelial cells and extracellular matrix proteins.

    Science.gov (United States)

    Jordan, Rachael P C; Williams, David W; Moran, Gary P; Coleman, David C; Sullivan, Derek J

    2014-04-01

    Candida albicans and Candida dubliniensis are very closely related pathogenic yeast species. Despite their close relationship, C. albicans is a far more successful colonizer and pathogen of humans. The purpose of this study was to determine if the disparity in the virulence of the two species is attributed to differences in their ability to adhere to human buccal epithelial cells (BECs) and/or extracellular matrix proteins. When grown overnight at 30°C in yeast extract peptone dextrose, genotype 1 C. dubliniensis isolates were found to be significantly more adherent to human BECs than C. albicans or C. dubliniensis genotypes 2-4 (P albicans to human BECs was observed, and C. dubliniensis genotype 1 and C. albicans adhered to BECs in significantly greater numbers than the other C. dubliniensis genotypes (P albicans to type I and IV collagen, fibronectin, laminin, vitronectin, and proline-rich peptides. These data suggest that C. albicans is not more adherent to epithelial cells or matrix proteins than C. dubliniensis and therefore other factors must contribute to the greater levels of virulence exhibited by C. albicans.

  19. Cannabinoid WIN-55,212-2 mesylate inhibits interleukin-1β induced matrix metalloproteinase and tissue inhibitor of matrix metalloproteinase expression in human chondrocytes.

    Science.gov (United States)

    Dunn, S L; Wilkinson, J M; Crawford, A; Le Maitre, C L; Bunning, R A D

    2014-01-01

    Interleukin-1β (IL-1β) is involved in the up-regulation of matrix metalloproteinases (MMPs) leading to cartilage degradation. Cannabinoids are anti-inflammatory and reduce joint damage in animal models of arthritis. This study aimed to determine a mechanism whereby the synthetic cannabinoid WIN-55,212-2 mesylate (WIN-55) may inhibit cartilage degradation. Effects of WIN-55 were studied on IL-1β stimulated production of MMP-3 and -13 and their inhibitors TIMP-1 and -2 in human chondrocytes. Chondrocytes were obtained from articular cartilage of patients undergoing total knee replacement. Chondrocytes were grown in monolayer and 3D alginate bead cultures. Real-time polymerase chain reaction (PCR) was used to determine the gene expression of MMP-3, -13, TIMP-1 and -2 and Enzyme Linked Immunosorbent Assay (ELISA) to measure the amount of MMP-3 and MMP-13 protein released into media. Immunocytochemistry was used to investigate the expression of cannabinoid receptors in chondrocyte cultures. Treatment with WIN-55 alone or in combination with IL-1β, decreased or abolished MMP-3, -13, TIMP-1 and -2 gene expression in human chondrocyte monolayer and alginate bead cultures in both a concentration and time dependent manner. WIN-55 treatment alone, and in combination with IL-1β, reduced MMP-3 and -13 protein production by chondrocytes cultured in alginate beads. Immunocytochemistry demonstrated the expression of cannabinoid receptors in chondrocyte cultures. Cannabinoid WIN-55 can reduce both basal and IL-1β stimulated gene and protein expression of MMP-3 and -13. However WIN-55 also decreased basal levels of TIMP-1 and -2 mRNA. These actions of WIN-55 suggest a mechanism by which cannabinoids may act to prevent cartilage breakdown in arthritis. Copyright © 2013 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.

  20. Human macrophages primed with angiogenic factors show dynamic plasticity, irrespective of extracellular matrix components

    NARCIS (Netherlands)

    Ploeger, Diana T. A.; van Putten, Sander M.; Koerts, Jasper A.; van Luyn, Marja J. A.; Harmsen, Martin C.

    2012-01-01

    Macrophages are important in inflammation as well as in tissue repair processes. They can be activated by various stimuli and classified into two major groups: M1 (classically activated) or M2 (alternatively activated). Inflammation, angiogenesis and matrix remodeling play a major role in tissue rep

  1. Interactions of human tenascin-X domains with dermal extracellular matrix molecules.

    NARCIS (Netherlands)

    Egging, D.; Berkmortel, F. van den; Taylor, G.; Bristow, J.; Schalkwijk, J.

    2007-01-01

    Tenascin-X (TNX) is a large 450 kDa extracellular matrix protein expressed in a variety of tissues including skin, joints and blood vessels. Deficiency of TNX causes a recessive form of Ehlers-Danlos syndrome characterized by joint hypermobility, skin fragility and hyperextensible skin. Skin of TNX

  2. Site controlled transgenic mice validating increased expression from human matrix metalloproteinase (MMP-1) promoter due to a naturally occurring SNP.

    Science.gov (United States)

    Coon, Charles I; Fiering, Steven; Gaudet, Justin; Wyatt, Colby A; Brinckerhoff, Constance E

    2009-09-01

    Matrix metalloproteinases (MMPs) comprise a family of more than 20 members, each with the ability to degrade components of the extracellular matrix. The interstitial collagenases have the unique capacity to degrade the stromal collagens, types I, II and III, the body's most abundant proteins. These collagenases include MMP-1, MMP-8, MMP-13 and MMP-14. MMP-1, with a very broad expression pattern, has major roles in mediating matrix destruction in many diseases. We have described a single nucleotide polymorphism (SNP) in the MMP-1 promoter that augments transcription. This SNP is the presence or absence of an extra guanine (G) at -1607 bp, which creates the sequence 5'-GGAA-3'(2G allele), and which is an ETS binding site. Compared to the 1G allele (5'-GAA-3'), the 2G SNP is associated with enhanced transcription of MMP-1 and increased enzymatic activity. Although murine systems are often used to model human diseases, mice have only distant homologues of human MMP-1. Therefore, we used a technique for the targeted insertion of a single copy of a gene at the HPRT locus to compare expression of the 1G and 2G alleles. We generated transgenic mice with -4372 bp of the human MMP-1 promoter containing either the 1G or 2G SNP in front of the lac Z (E.coli ss-galactosidase) gene. We measured the relative expression of the transgenes in vitro in embryonic stem (ES) cells and in fibroblasts derived from embryonic mice. Our data show modest constitutive expression of ss-galactosidase mRNA and protein from these alleles, with the 2G allele more transcriptionally active than the 1G allele. We conclude that these mice represent a model for integration of a single copy of the human MMP-1 promoter into the murine genome, and could be used to study MMP-1 gene expression in a murine system.

  3. Expression of extracellular matrix components and related growth factors in human tendon and muscle after acute exercise

    DEFF Research Database (Denmark)

    Heinemeier, K M; Bjerrum, S S; Schjerling, P

    2013-01-01

    the patellar tendon and vastus lateralis muscle of each leg at 2 (n = 10), 6 (n = 11), or 26 h (n = 10) after exercise. Levels of messenger ribonucleic acid mRNA for collagens, noncollagenous matrix proteins, and growth factors were measured with real-time reverse transcription polymerase chain reaction......Acute kicking exercise induces collagen synthesis in both tendon and muscle in humans, but it is not known if this relates to increased collagen transcription and if other matrix genes are regulated. Young men performed 1 h of one-leg kicking at 67% of max workload. Biopsies were taken from....... In tendon, gene expression was unchanged except for a decrease in insulin-like growth factor-IEa (IGF-IEa; P ...

  4. Differential Expression of Matrix Metalloproteases in Human Fibroblasts with Different Origins

    Directory of Open Access Journals (Sweden)

    Diana Lindner

    2012-01-01

    Full Text Available Fibroblasts are widely distributed cells and are responsible for the deposition of extracellular matrix (ECM components but also secrete ECM-degrading matrix metalloproteases. A finely balanced equilibrium between deposition and degradation of ECM is essential for structural integrity of tissues. In the past, fibroblasts have typically been understood as a uniform cell population with comparable functions regardless of their origin. Here, we determined growth curves of fibroblasts derived from heart, skin, and lung and clearly show the lowest proliferation rate for cardiac fibroblasts. Furthermore, we examined basal expression levels of collagen and different MMPs in these three types of fibroblasts and compared these concerning their site of origin. Interestingly, we found major differences in basal mRNA expression especially for MMP1 and MMP3. Moreover, we treated fibroblasts with TNF-α and observed different alterations under these proinflammatory conditions. In conclusion, fibroblasts show different properties in proliferation and MMP expression regarding their originated tissue.

  5. Repeated short climatic change affects the epidermal differentiation program and leads to matrix remodeling in a human organotypic skin model

    Science.gov (United States)

    Boutrand, Laetitia-Barbollat; Thépot, Amélie; Muther, Charlotte; Boher, Aurélie; Robic, Julie; Guéré, Christelle; Vié, Katell; Damour, Odile; Lamartine, Jérôme

    2017-01-01

    Human skin is subject to frequent changes in ambient temperature and humidity and needs to cope with these environmental modifications. To decipher the molecular response of human skin to repeated climatic change, a versatile model of skin equivalent subject to “hot–wet” (40°C, 80% relative humidity [RH]) or “cold–dry” (10°C, 40% RH) climatic stress repeated daily was used. To obtain an exhaustive view of the molecular mechanisms elicited by climatic change, large-scale gene expression DNA microarray analysis was performed and modulated function was determined by bioinformatic annotation. This analysis revealed several functions, including epidermal differentiation and extracellular matrix, impacted by repeated variations in climatic conditions. Some of these molecular changes were confirmed by histological examination and protein expression. Both treatments (hot–wet and cold–dry) reduced the expression of genes encoding collagens, laminin, and proteoglycans, suggesting a profound remodeling of the extracellular matrix. Strong induction of the entire family of late cornified envelope genes after cold–dry exposure, confirmed at protein level, was also observed. These changes correlated with an increase in epidermal differentiation markers such as corneodesmosin and a thickening of the stratum corneum, indicating possible implementation of defense mechanisms against dehydration. This study for the first time reveals the complex pattern of molecular response allowing adaption of human skin to repeated change in its climatic environment.

  6. Dentin matrix protein 1 and dentin sialophosphoprotein in human sound and carious teeth: an immunohistochemical and colorimetric assay

    Directory of Open Access Journals (Sweden)

    D. Martini

    2013-10-01

    Full Text Available Dentin matrix protein 1 (DMP1 and dentin sialophosphoprotein (DSPP are extracellular matrix proteins produced by odontoblasts involved in the dentin mineralization. The aim this study was to compare the distribution of DMP1 and DSPP in human sound dentin vs human sclerotic dentin. Sixteen sound and sixteen carious human molars were selected, fixed in paraformaldehyde and processed for immunohistochemical detection of DMP1 and DSPP by means of light microscopy, transmission electron microscopy (TEM and high-resolution field emission in-lens scanning electron microscopy (FEI-SEM. Specimens were submitted to a pre-embedding or a post-embedding immunolabeling technique using primary antibodies anti DMP1 and anti-DSPP and gold-conjugated secondary antibodies. Other samples were processed for the detection of DMP1 and DSPP levels. Dentin from these samples was mechanically fractured to powder, then a protein extraction and a protein level detection assay were performed. DMP1 and DSPP were more abundant in carious than in sound samples. Immunohistochemical analyses in sclerotic dentin disclosed a high expression of DMP1 and DSPP inside the tubules, suggesting an active biomineralization of dentin by odontoblasts. Furthermore, the detection of small amounts of these proteins inside the tubules far from the carious lesion, as shown in the present study, is consistent with the hypothesis of a preventive defense of all dentin after a noxious stimulus has undermined the tooth.

  7. Dentin matrix protein 1 and dentin sialophosphoprotein in human sound and carious teeth: an immunohistochemical and colorimetric assay.

    Science.gov (United States)

    Martini, D; Trirè, A; Breschi, L; Mazzoni, A; Teti, G; Falconi, M; Ruggeri, A

    2013-10-29

    Dentin matrix protein 1 (DMP1) and dentin sialophosphoprotein (DSPP) are extracellular matrix proteins produced by odontoblasts involved in the dentin mineralization. The aim this study was to compare the distribution of DMP1 and DSPP in human sound dentin vs human sclerotic dentin. Sixteen sound and sixteen carious human molars were selected, fixed in paraformaldehyde and processed for immunohistochemical detection of DMP1 and DSPP by means of light microscopy, transmission electron microscopy (TEM) and high-resolution field emission in-lens scanning electron microscopy (FEI-SEM). Specimens were submitted to a pre-embedding or a post-embedding immunolabeling technique using primary antibodies anti DMP1 and anti-DSPP and gold-conjugated secondary antibodies. Other samples were processed for the detection of DMP1 and DSPP levels. Dentin from these samples was mechanically fractured to powder, then a protein extraction and a protein level detection assay were performed. DMP1 and DSPP were more abundant in carious than in sound samples. Immunohistochemical analyses in sclerotic dentin disclosed a high expression of DMP1 and DSPP inside the tubules, suggesting an active biomineralization of dentin by odontoblasts. Furthermore, the detection of small amounts of these proteins inside the tubules far from the carious lesion, as shown in the present study, is consistent with the hypothesis of a preventive defense of all dentin after a noxious stimulus has undermined the tooth.

  8. Dentin Matrix Protein 1 and Dentin Sialophosphoprotein in Human Sound and Carious Teeth: An Immunohistochemical and Colorimetric Assay

    Science.gov (United States)

    Martini, D.; Trirè, A.; Breschi, L.; Mazzoni, A.; Orsini, G.; Teti, G.; Falconi, M.; Ruggeri, A.

    2013-01-01

    Dentin matrix protein 1 (DMP1) and dentin sialophosphoprotein (DSPP) are extracellular matrix proteins produced by odontoblasts involved in the dentin mineralization. The aim this study was to compare the distribution of DMP1 and DSPP in human sound dentin vs human sclerotic dentin. Sixteen sound and sixteen carious human molars were selected, fixed in paraformaldehyde and processed for immunohistochemical detection of DMP1 and DSPP by means of light microscopy, transmission electron microscopy (TEM) and high-resolution field emission in-lens scanning electron microscopy (FEI-SEM). Specimens were submitted to a pre-embedding or a post-embedding immunolabeling technique using primary antibodies anti DMP1 and anti-DSPP and gold-conjugated secondary antibodies. Other samples were processed for the detection of DMP1 and DSPP levels. Dentin from these samples was mechanically fractured to powder, then a protein extraction and a protein level detection assay were performed. DMP1 and DSPP were more abundant in carious than in sound samples. Immunohistochemical analyses in sclerotic dentin disclosed a high expression of DMP1 and DSPP inside the tubules, suggesting an active biomineralization of dentin by odontoblasts. Furthermore, the detection of small amounts of these proteins inside the tubules far from the carious lesion, as shown in the present study, is consistent with the hypothesis of a preventive defense of all dentin after a noxious stimulus has undermined the tooth. PMID:24441185

  9. Guided bone regeneration using acellular bovine pericardium in a rabbit mandibular model: in-vitro and in-vivo studies.

    Science.gov (United States)

    Bai, M; Zhang, T; Ling, T; Zhou, Z; Xie, H; Zhang, W; Hu, G; Jiang, C; Li, M; Feng, B; Wu, H

    2014-08-01

    To investigate the feasibility of acellular bovine pericardium (BP) for guided bone regeneration (GBR) in vitro and in vivo. The success of GBR relies on the fact that various cellular components possess different migration rates into the defect site and that a barrier membrane plays a significant role in these processes. BP membrane was isolated and decellularized using an enzymatic method. The microarchitecture, mechanical properties, cytotoxicity and cell chemotaxis properties of the acellular BP were evaluated in vitro, and the in-vivo efficacy of the acellular BP was also investigated in a rabbit mandibular model. The acellular BP membrane possessed an interconnected fibrous structure. Glutaraldehyde (GA) treatment was efficient for enhancement of the mechanical properties of the acellular BP bur and resulted in negligible cytotoxicity. After 16 wk, standardized osseous defects created in the rabbit mandible, and covered with acellular BP, were associated with an enhanced deposition of mineralized tissue when compared with defects left to spontaneous healing. GA-treated acellular BP is promising as a barrier membrane for GBR for further in-vivo and clinical studies. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  10. Human lung fibroblast-derived matrix facilitates vascular morphogenesis in 3D environment and enhances skin wound healing.

    Science.gov (United States)

    Du, Ping; Suhaeri, Muhammad; Ha, Sang Su; Oh, Seung Ja; Kim, Sang-Heon; Park, Kwideok

    2017-05-01

    Extracellular matrix (ECM) is crucial to many aspects of vascular morphogenesis and maintenance of vasculature function. Currently the recapitulation of angiogenic ECM microenvironment is still challenging, due mainly to its diverse components and complex organization. Here we investigate the angiogenic potential of human lung fibroblast-derived matrix (hFDM) in creating a three-dimensional (3D) vascular construct. hFDM was obtained via decellularization of in vitro cultured human lung fibroblasts and analyzed via immunofluorescence staining and ELISA, which detect multiple ECM macromolecules and angiogenic growth factors (GFs). Human umbilical vein endothelial cells (HUVECs) morphology was more elongated and better proliferative on hFDM than on gelatin-coated substrate. To prepare 3D construct, hFDM is collected, quantitatively analyzed, and incorporated in collagen hydrogel (Col) with HUVECs. Capillary-like structure (CLS) formation at 7day was significantly better with the groups containing higher doses of hFDM compared to the Col group (control). Moreover, the group (Col/hFDM/GFs) with both hFDM and angiogenic GFs (VEGF, bFGF, SDF-1) showed the synergistic activity on CLS formation and found much larger capillary lumen diameters with time. Further analysis of hFDM via angiogenesis antibody array kit reveals abundant biochemical cues, such as angiogenesis-related cytokines, GFs, and proteolytic enzymes. Significantly up-regulated expression of VE-cadherin and ECM-specific integrin subunits was also noticed in Col/hFDM/GFs. In addition, transplantation of Col/hFMD/GFs with HUVECs in skin wound model presents more effective re-epithelialization, many regenerated hair follicles, better transplanted cells viability, and advanced neovascularization. We believe that current system is a very promising platform for 3D vasculature construction in vitro and for cell delivery toward therapeutic applications in vivo. Functional 3D vasculature construction in vitro is still

  11. YKL-40, a mammalian member of the chitinase family, is a matrix protein of specific granules in human neutrophils

    DEFF Research Database (Denmark)

    Volck, B; Price, P A; Johansen, J S;

    1998-01-01

    YKL-40, also called human cartilage glycoprotein-39 (HC gp-39), is a member of family 18 glycosyl hydrolases. YKL-40 is secreted by chondrocytes, synovial cells, and macrophages, and recently it has been reported that YKL-40 has a role as an autoantigen in rheumatoid arthritis (RA). The function...... of YKL-40 is unknown, but the pattern of its expression in normal and disease states suggests that it could function in remodeling or degradation of the extracellular matrix. High levels of YKL-40 are found in synovial fluid from patients with active RA. Neutrophils are abundant in synovial fluid...

  12. Release of tensile strain on engineered human tendon tissue disturbs cell adhesions, changes matrix architecture, and induces an inflammatory phenotype

    DEFF Research Database (Denmark)

    Bayer, Monika L; Schjerling, Peter; Herchenhan, Andreas

    2014-01-01

    -inflammatory mediators and tendon phenotypic specific molecules, in an in vitro model where tendon-like tissue was engineered from human tendon cells. Tissue sampling was performed 1, 2, 4 and 6 days after surgical de-tensioning of the tendon construct. When tensile stimulus was removed, integrin type collagen receptors...... were upregulated. Stimulation with the cytokine TGF-β1 had distinct effects on some tendon-related genes in both tensioned and de-tensioned tissue. These findings indicate an important role of mechanical loading for cellular and matrix responses in tendon, including that loss of tension leads...

  13. Adaptive bone formation in acellular vertebrae of sea bass (Dicentrarchus labrax L.)

    NARCIS (Netherlands)

    Kranenbarg, S.; Cleynenbreugel, van T.; Schipper, H.; Leeuwen, van J.L.

    2005-01-01

    Mammalian bone is an active tissue in which osteoblasts and osteoclasts balance bone mass. This process of adaptive modelling and remodelling is probably regulated by strain-sensing osteocytes. Bone of advanced teleosts is acellular yet, despite the lack of osteocytes, it is capable of an adaptive

  14. Biomechanical properties of acellular sciatic nerves treated with a modified chemical method

    Institute of Scientific and Technical Information of China (English)

    Xinlong Ma; Zhao Yang; Xiaolei Sun; Jianxiong Ma; Xiulan Li; Zhenzhen Yuan; Yang Zhang; Honggang Guo

    2011-01-01

    Nerve grafts are able to adapt to surrounding biomechanical environments if the nerve graft itself exhibits appropriate biomechanical properties (load, elastic modulus, etc.). The present study was designed to determine the differences in biomechanical properties between fresh and chemically acellularized sciatic nerve grafts. Two different chemical methods were used to establish acellular nerve grafts. The nerve was chemically extracted in the Sondell method with a combination of Triton X-100 (nonionic detergent) and sodium deoxycholate (anionic detergent), and in the modified method with a combination of Triton X-200 (anionic detergent), sulfobetaine-10 (SB-10, amphoteric detergents), and sulfobetaine-16 (SB-16, amphoteric detergents). Following acellularization, hematoxylin-eosin staining and scanning electron microscopy demonstrated that the effect of acellularization via the modified method was similar to the traditional Sondell method. However, effects of demyelination and nerve fiber tube integrity were superior to the traditional Sondell method. Biomechanical testing showed that peripheral nerve graft treated using the chemical method resulted in decreased biomechanical properties (ultimate load, ultimate stress, ultimate strain, and mechanical work to fracture) compared with fresh nerves, but the differences had no statistical significance (P > 0.05). These results demonstrated no significant effect on biomechanical properties of nerves treated using the chemical method. In conclusion, nerve grafts treated via the modified method removed Schwann cells, preserved neural structures, and ensured biomechanical properties of the nerve graft, which could be more appropriate for implantation studies.

  15. Integrin-mediated interactions with extracellular matrix proteins for nucleus pulposus cells of the human intervertebral disc.

    Science.gov (United States)

    Bridgen, D T; Gilchrist, C L; Richardson, W J; Isaacs, R E; Brown, C R; Yang, K L; Chen, J; Setton, L A

    2013-10-01

    The extracellular matrix (ECM) of the human intervertebral disc is rich in molecules that interact with cells through integrin-mediated attachments. Porcine nucleus pulposus (NP) cells have been shown to interact with laminin (LM) isoforms LM-111 and LM-511 through select integrins that regulate biosynthesis and cell attachment. Since human NP cells lose many phenotypic characteristics with age, attachment and interaction with the ECM may be altered. Expression of LM-binding integrins was quantified for human NP cells using flow cytometry. The cell-ECM attachment mechanism was determined by quantifying cell attachment to LM-111, LM-511, or type II collagen after functionally blocking specific integrin subunits. Human NP cells express integrins β1, α3, and α5, with over 70% of cells positive for each subunit. Blocking subunit β1 inhibited NP cell attachment to all substrates. Blocking subunits α1, α2, α3, and α5 simultaneously, but not individually, inhibits NP cell attachment to laminins. While integrin α6β1 mediated porcine NP cell attachment to LM-111, we found integrins α3, α5, and β1 instead contributed to human NP cell attachment. These findings identify integrin subunits that may mediate interactions with the ECM for human NP cells and could be used to promote cell attachment, survival, and biosynthesis in cell-based therapeutics.

  16. Release of Tensile Strain on Engineered Human Tendon Tissue Disturbs Cell Adhesions, Changes Matrix Architecture, and Induces an Inflammatory Phenotype

    Science.gov (United States)

    Bayer, Monika L.; Schjerling, Peter; Herchenhan, Andreas; Zeltz, Cedric; Heinemeier, Katja M.; Christensen, Lise; Krogsgaard, Michael; Gullberg, Donald; Kjaer, Michael

    2014-01-01

    Mechanical loading of tendon cells results in an upregulation of mechanotransduction signaling pathways, cell-matrix adhesion and collagen synthesis, but whether unloading removes these responses is unclear. We investigated the response to tension release, with regard to matrix proteins, pro-inflammatory mediators and tendon phenotypic specific molecules, in an in vitro model where tendon-like tissue was engineered from human tendon cells. Tissue sampling was performed 1, 2, 4 and 6 days after surgical de-tensioning of the tendon construct. When tensile stimulus was removed, integrin type collagen receptors showed a contrasting response with a clear drop in integrin subunit α11 mRNA and protein expression, and an increase in α2 integrin mRNA and protein levels. Further, specific markers for tendon cell differentiation declined and normal tendon architecture was disturbed, whereas pro-inflammatory molecules were upregulated. Stimulation with the cytokine TGF-β1 had distinct effects on some tendon-related genes in both tensioned and de-tensioned tissue. These findings indicate an important role of mechanical loading for cellular and matrix responses in tendon, including that loss of tension leads to a decrease in phenotypical markers for tendon, while expression of pro-inflammatory mediators is induced. PMID:24465881

  17. Influence of culture conditions and extracellular matrix alignment on human mesenchymal stem cells invasion into decellularized engineered tissues.

    Science.gov (United States)

    Weidenhamer, Nathan K; Moore, Dusty L; Lobo, Fluvio L; Klair, Nathaniel T; Tranquillo, Robert T

    2015-05-01

    The variables that influence the in vitro recellularization potential of decellularized engineered tissues, such as cell culture conditions and scaffold alignment, have yet to be explored. The goal of this work was to explore the influence of insulin and ascorbic acid and extracellular matrix (ECM) alignment on the recellularization of decellularized engineered tissue by human mesenchymal stem cells (hMSCs). Aligned and non-aligned tissues were created by specifying the geometry and associated mechanical constraints to fibroblast-mediated fibrin gel contraction and remodelling using circular and C-shaped moulds. Decellularized tissues (matrices) of the same alignment were created by decellularization with detergents. Ascorbic acid promoted the invasion of hMSCs into the matrices due to a stimulated increase in motility and proliferation. Invasion correlated with hyaluronic acid secretion, α-smooth muscle actin expression and decreased matrix thickness. Furthermore, hMSCs invasion into aligned and non-aligned matrices was not different, although there was a difference in cell orientation. Finally, we show that hMSCs on the matrix surface appear to differentiate toward a smooth muscle cell or myofibroblast phenotype with ascorbic acid treatment. These results inform the strategy of recellularizing decellularized engineered tissue with hMSCs.

  18. The mechanical properties of human adipose tissues and their relationships to the structure and composition of the extracellular matrix.

    Science.gov (United States)

    Alkhouli, Nadia; Mansfield, Jessica; Green, Ellen; Bell, James; Knight, Beatrice; Liversedge, Neil; Tham, Ji Chung; Welbourn, Richard; Shore, Angela C; Kos, Katarina; Winlove, C Peter

    2013-12-01

    Adipose tissue (AT) expansion in obesity is characterized by cellular growth and continuous extracellular matrix (ECM) remodeling with increased fibrillar collagen deposition. It is hypothesized that the matrix can inhibit cellular expansion and lipid storage. Therefore, it is important to fully characterize the ECM's biomechanical properties and its interactions with cells. In this study, we characterize and compare the mechanical properties of human subcutaneous and omental tissues, which have different physiological functions. AT was obtained from 44 subjects undergoing surgery. Force/extension and stress/relaxation data were obtained. The effects of osmotic challenge were measured to investigate the cellular contribution to tissue mechanics. Tissue structure and its response to tensile strain were determined using nonlinear microscopy. AT showed nonlinear stress/strain characteristics of up to a 30% strain. Comparing paired subcutaneous and omental samples (n = 19), the moduli were lower in subcutaneous: initial 1.6 ± 0.8 (means ± SD) and 2.9 ± 1.5 kPa (P = 0.001), final 11.7 ± 6.4 and 32 ± 15.6 kPa (P matrix fibers. These results suggest that subcutaneous AT has greater capacity for expansion and recovery from mechanical deformation than omental AT.

  19. Molecular docking analysis of selected Clinacanthus nutans constituents as xanthine oxidase, nitric oxide synthase, human neutrophil elastase, matrix metalloproteinase 2, matrix metalloproteinase 9 and squalene synthase inhibitors

    Directory of Open Access Journals (Sweden)

    Radhakrishnan Narayanaswamy

    2016-01-01

    Full Text Available Background: Clinacanthus nutans (Burm. f. Lindau has gained popularity among Malaysians as a traditional plant for anti-inflammatory activity. Objective: This prompted us to carry out the present study on a selected 11 constituents of C. nutans which are clinacoside A–C, cycloclinacoside A1, shaftoside, vitexin, orientin, isovitexin, isoorientin, lupeol and β-sitosterol. Materials and Methods: Selected 11 constituents of C. nutans were evaluated on the docking behavior of xanthine oxidase (XO, nitric oxide synthase (NOS, human neutrophil elastase (HNE, matrix metalloproteinase (MMP 2 and 9, and squalene synthase (SQS using Discovery Studio Version 3.1. Also, molecular physicochemical, bioactivity, absorption, distribution, metabolism, excretion, and toxicity (ADMET, and toxicity prediction by computer assisted technology analyzes were also carried out. Results: The molecular physicochemical analysis revealed that four ligands, namely clinacoside A–C and cycloclinacoside A1 showed nil violations and complied with Lipinski's rule of five. As for the analysis of bioactivity, all the 11 selected constituents of C. nutans exhibited active score (>0 toward enzyme inhibitors descriptor. ADMET analysis showed that the ligands except orientin and isoorientin were predicted to have Cytochrome P4502D6 inhibition effect. Docking studies and binding free energy calculations revealed that clinacoside B exhibited the least binding energy for the target enzymes except for XO and SQS. Isovitexin and isoorientin showed the potentials in the docking and binding with all of the six targeted enzymes, whereas vitexin and orientin docked and bound with only NOS and HNE. Conclusion: This present study has paved a new insight in understanding these 11 C. nutans ligands as potential inhibitors against XO, NOS, HNE, MMP 2, MMP 9, and SQS.

  20. TCDD induces dermal accumulation of keratinocyte-derived matrix metalloproteinase-10 in an organotypic model of human skin

    Energy Technology Data Exchange (ETDEWEB)

    De Abrew, K. Nadira [Molecular and Environmental Toxicology Center, University of Wisconsin—Madison, Madison, WI 53706 (United States); Thomas-Virnig, Christina L.; Rasmussen, Cathy A. [Department of Pathology, University of Wisconsin—Madison, Madison, WI 53706 (United States); Bolterstein, Elyse A. [Molecular and Environmental Toxicology Center, University of Wisconsin—Madison, Madison, WI 53706 (United States); Schlosser, Sandy J. [Department of Pathology, University of Wisconsin—Madison, Madison, WI 53706 (United States); Allen-Hoffmann, B. Lynn, E-mail: blallenh@wisc.edu [Molecular and Environmental Toxicology Center, University of Wisconsin—Madison, Madison, WI 53706 (United States); Department of Pathology, University of Wisconsin—Madison, Madison, WI 53706 (United States)

    2014-05-01

    The epidermis of skin is the first line of defense against the environment. A three dimensional model of human skin was used to investigate tissue-specific phenotypes induced by the environmental contaminant, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Continuous treatment of organotypic cultures of human keratinocytes with TCDD resulted in intracellular spaces between keratinocytes of the basal and immediately suprabasal layers as well as thinning of the basement membrane, in addition to the previously reported hyperkeratinization. These tissue remodeling events were preceded temporally by changes in expression of the extracellular matrix degrading enzyme, matrix metalloproteinase-10 (MMP-10). In organotypic cultures MMP-10 mRNA and protein were highly induced following TCDD treatment. Q-PCR and immunoblot results from TCDD-treated monolayer cultures, as well as indirect immunofluorescence and immunoblot analysis of TCDD-treated organotypic cultures, showed that MMP-10 was specifically contributed by the epidermal keratinocytes but not the dermal fibroblasts. Keratinocyte-derived MMP-10 protein accumulated over time in the dermal compartment of organotypic cultures. TCDD-induced epidermal phenotypes in organotypic cultures were attenuated by the keratinocyte-specific expression of tissue inhibitor of metalloproteinase-1, a known inhibitor of MMP-10. These studies suggest that MMP-10 and possibly other MMP-10-activated MMPs are responsible for the phenotypes exhibited in the basement membrane, the basal keratinocyte layer, and the cornified layer of TCDD-treated organotypic cultures. Our studies reveal a novel mechanism by which the epithelial–stromal microenvironment is altered in a tissue-specific manner thereby inducing structural and functional pathology in the interfollicular epidermis of human skin. - Highlights: • TCDD causes hyperkeratosis and basement membrane changes in a model of human skin. • TCDD induces MMP-10 expression in organotypic cultures

  1. Human umbilical cord matrix mesenchymal stem cells suppress the growth of breast cancer by expression of tumor suppressor genes.

    Directory of Open Access Journals (Sweden)

    Naomi Ohta

    Full Text Available Human and rat umbilical cord matrix mesenchymal stem cells (UCMSC possess the ability to control the growth of breast carcinoma cells. Comparative analyses of two types of UCMSC suggest that rat UCMSC-dependent growth regulation is significantly stronger than that of human UCMSC. Their different tumoricidal abilities were clarified by analyzing gene expression profiles in the two types of UCMSC. Microarray analysis revealed differential gene expression between untreated naïve UCMSC and those co-cultured with species-matched breast carcinoma cells. The analyses screened 17 differentially expressed genes that are commonly detected in both human and rat UCMSC. The comparison between the two sets of gene expression profiles identified two tumor suppressor genes, adipose-differentiation related protein (ADRP and follistatin (FST, that were specifically up-regulated in rat UCMSC, but down-regulated in human UCMSC when they were co-cultured with the corresponding species' breast carcinoma cells. Over-expression of FST, but not ADRP, in human UCMSC enhanced their ability to suppress the growth of MDA-231 cells. The growth of MDA-231 cells was also significantly lower when they were cultured in medium conditioned with FST, but not ADRP over-expressing human UCMSC. In the breast carcinoma lung metastasis model generated with MDA-231 cells, systemic treatment with FST-over-expressing human UCMSC significantly attenuated the tumor burden. These results suggest that FST may play an important role in exhibiting stronger tumoricidal ability in rat UCMSC than human UCMSC and also implies that human UCMSC can be transformed into stronger tumoricidal cells by enhancing tumor suppressor gene expression.

  2. PRDM14 Promotes the Migration of Human Non-small Cell Lung Cancer Through Extracellular Matrix Degradation in vitro

    Institute of Scientific and Technical Information of China (English)

    Hong-Xia Bi; Han-Bing Shi; Ting Zhang; Ge Cui

    2015-01-01

    Background:As a novel molecular markerof non-small cell lung cancer (NSCLC),PRDI-BF1 and RIZ homology domain containing protein 14 (PRDM 14) is over-expressed in NSCLC tumor tissues.Extracellular matrix degradation mediated by the balance between matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinases (TIMPs) is one of the most important mechanism in lung cancer metastasis.This study aimed to determine if PRDM 14 promoted the migration of NSCLC cells through extracellular matrix degradation mediated by change of MMP/TIMP expression.Methods:The expression of PRDM14 was down-regulated in human cell line A 549 after transfection with lentiviral vector-mediated short-hairpin ribonucleic acids (shRNAs) which targeted the PRDM 14 promoter.Cellular migration ofshRNA-infected cells was detected by a scratch wound healing assay and transwell cell migration assay.Expression levels of MMP1,MMP2,TIMP1,and TIMP2 were measured by quantitative real-time polymerase chain reaction (RT-PCR).Results:Migration ofPRDM 14-shRNA-infected cells was significantly inhibited relative to control cells as measured by the scratch wound healing (P < 0.05) and transwell cell migration assays (P < 0.01).The expression of MMP1 in A549 cells infected by PRDM14-shRNA was down-regulated significantly (P < 0.01),whereas the expression of TIMPl and TIMP2 was up-regulated significantly (P < 0.01).Conclusions:PRDM 14 accelerates A549 cells migration in vitro through extracellular matrix degradation.PRDM 14 is considered as a potential therapeutic target in metastatic NSCLC.

  3. Resorbable extracellular matrix grafts in urologic reconstruction

    Directory of Open Access Journals (Sweden)

    Richard A. Santucci

    2005-06-01

    Full Text Available PURPOSE: There is an increasingly large body of literature concerning tissue-engineering products that may be used in urology. Some of these are quite complex (such as multilayer patient-specific cell-seeded implants yet the most simple and successful products to date are also the most uncomplicated: resorbable acellular extra-cellular matrices (ECMs harvested from animals. ECMs have been used in a variety of difficult urologic reconstruction problems, and this review is intended to summarize this complex literature for the practicing urologist. METHODS: Medline search of related terms such as "SIS, small intestinal submucosa, ECM, extracellular matrix, acellular matrix and urologic reconstruction". Manuscripts missed in the initial search were taken from the bibliographies of the primary references. RESULTS: Full review of potential clinical uses of resorbable extra-cellular matrices in urologic reconstruction. CONCLUSIONS: Currently, the "state of the art" in tissue engineering solutions for urologic reconstruction means resorbable acellular xenograft matrices. They show promise when used as a pubovaginal sling or extra bolstering layers in ureteral or urethral repairs, although recent problems with inflammation following 8-ply pubovaginal sling use and failures after 1- and 4-ply SIS repair of Peyronie's disease underscore the need for research before wide adoption. Preliminary data is mixed concerning the potential for ECM urethral patch graft, and more data is needed before extended uses such as bladder augmentation and ureteral replacement are contemplated. The distant future of ECMs in urology likely will include cell-seeded grafts with the eventual hope of producing "off the shelf" replacement materials. Until that day arrives, ECMs only fulfill some of the requirements for the reconstructive urologist.

  4. Matrix stiffness regulation of integrin-mediated mechanotransduction during osteogenic differentiation of human mesenchymal stem cells.

    Science.gov (United States)

    Shih, Yu-Ru V; Tseng, Kuo-Fung; Lai, Hsiu-Yu; Lin, Chi-Hung; Lee, Oscar K

    2011-04-01

    Mesenchymal stem cells (MSCs) cultured on extracellular matrices with different stiffness have been shown to possess diverse lineage commitment owing to the extracellular mechanical stimuli sensed by the cells. The aim of this study was to further delineate how matrix stiffness affects intracellular signaling through the mechanotransducers Rho kinase (ROCK) and focal adhesion kinase (FAK) and subsequently regulates the osteogenic phenotype of MSCs. MSCs were cultured in osteogenic medium on tunable polyacrylamide hydrogels coated with type I collagen with elasticities corresponding to Young's modulus of 7.0 ± 1.2 and 42.1 ± 3.2 kPa. Osteogenic differentiation was increased on stiffer matrices, as evident by type I collagen, osteocalcin, and Runx2 gene expressions and alizarin red S staining for mineralization. Western blot analysis demonstrated an increase in kinase activities of ROCK, FAK, and ERK1/2 on stiffer matrices. Inhibition of FAK, an important mediator of osteogenic differentiation, and inhibition of ROCK, a known mechanotransducer of matrix stiffness during osteogenesis, resulted in decreased expression of osteogenic markers during osteogenic induction. In addition, FAK affects osteogenic differentiation through ERK1/2, whereas ROCK regulates both FAK and ERK1/2. Furthermore, α(2)-integrin was upregulated on stiffer matrices during osteogenic induction, and its knockdown by siRNA downregulated the osteogenic phenotype through ROCK, FAK, and ERK1/2. Taken together, our results provide evidence that the matrix rigidity affects the osteogenic outcome of MSCs through mechanotransduction events that are mediated by α(2)-integrin.

  5. Expression of matrix metalloproteinases 2 and 9 in human gastric cancer and superficial gastritis

    Institute of Scientific and Technical Information of China (English)

    Clara; Luz; Sampieri; Sol; de; la; Pea; Mariana; Ochoa-Lara; Roberto; Zenteno-Cuevas; Kenneth; León-Córdoba

    2010-01-01

    AIM:To assess expression of matrix metalloproteinases 2(MMP2)and MMP9 in gastric cancer,superficial gastritis and normal mucosa,and to measure metalloproteinase activity.METHODS:MMP2 and MMP9 mRNA expression was determined by quantitative real-time polymerase chain reaction.Normalization was carried out using three different factors.Proteins were analyzed by quantitative gelatin zymography(qGZ).RESULTS:18S ribosomal RNA(18SRNA)was very highly expressed,while hypoxanthine ribosyltransferase-1(HPRT-1)was mode...

  6. Light and electron microscopic study of epithelial cells from the human oviduct and uterus subcultured on extracellular matrix gel.

    Science.gov (United States)

    Eslaminejad, Mohamadreza Baghaban; Valojerdi, Mojtaba Rezazadeh; Ashtiani, Saeed Kazemi; Eftekhari-Yazdi, Poopak

    2007-06-01

    To investigate the structure of epithelial cells from the human oviduct and uterus on extracellular matrix (ECM) gel in the first passage. Human oviducts and endometrial tissues were obtained from patients undergoing total hysterectomy; the epithelial cells, having been isolated by enzyme digestion, were cultured on polystyrene plastic surfaces. The epithelial nature of the cells was confirmed by immunocytochemistry, and their morphology was examined by microscopy. Cells of an epithelial nature were then trypsinized and cultured on an ECM gel-coated filter insert for 5 days. The cells, in parallel with the tissues, were subsequently prepared for transmission electron microscopy. Plastic-cultured cells had no sign of differentiation and appeared as elongated spindle cells in sections. These cells looked columnar and highly polarized after being cultured on ECM gel surfaces. They were similar to epithelial cells from the corresponding tissue fragment. Cultured on ECM gel, the ciliated epithelial cells of human oviducts appeared ultrastructurally similar to glandular cells from the human uterus. Cilia did not form under culture conditions. It seems that human uterine and oviduct epithelial cells can acquire polarized morphology and differentiated states on ECM gel after having lost it on plastic surfaces and that ECM gel by itself is not enough to induce cilia formation in culture.

  7. Cultivation of corneal endothelial cells on a pericellular matrix prepared from human decidua-derived mesenchymal cells.

    Directory of Open Access Journals (Sweden)

    Ryohei Numata

    Full Text Available The barrier and pump functions of the corneal endothelium are essential for the maintenance of corneal transparency. Although corneal transplantation is the only current therapy for treating corneal endothelial dysfunction, the potential of tissue-engineering techniques to provide highly efficient and less invasive therapy in comparison to corneal transplantation has been highly anticipated. However, culturing human corneal endothelial cells (HCECs is technically difficult, and there is no established culture protocol. The aim of this study was to investigate the feasibility of using a pericellular matrix prepared from human decidua-derived mesenchymal cells (PCM-DM as an animal-free substrate for HCEC culture for future clinical applications. PCM-DM enhanced the adhesion of monkey CECs (MCECs via integrin, promoted cell proliferation, and suppressed apoptosis. The HCECs cultured on the PCM-DM showed a hexagonal morphology and a staining profile characteristic of Na⁺/K⁺-ATPase and ZO-1 at the plasma membrane in vivo, whereas the control HCECs showed a fibroblastic phenotype. The cell density of the cultured HCECs on the PCM-DM was significantly higher than that of the control cells. These results indicate that PCM-DM provides a feasible xeno-free matrix substrate and that it offers a viable in vitro expansion protocol for HCECs while maintaining cellular functions for use as a subsequent clinical intervention for tissue-engineered based therapy of corneal endothelial dysfunction.

  8. Altered expression of mitochondrial and extracellular matrix genes in the heart of human fetuses with chromosome 21 trisomy

    Directory of Open Access Journals (Sweden)

    Olla Carlo

    2007-08-01

    Full Text Available Abstract Background The Down syndrome phenotype has been attributed to overexpression of chromosome 21 (Hsa21 genes. However, the expression profile of Hsa21 genes in trisomic human subjects as well as their effects on genes located on different chromosomes are largely unknown. Using oligonucleotide microarrays we compared the gene expression profiles of hearts of human fetuses with and without Hsa21 trisomy. Results Approximately half of the 15,000 genes examined (87 of the 168 genes on Hsa21 were expressed in the heart at 18–22 weeks of gestation. Hsa21 gene expression was globally upregulated 1.5 fold in trisomic samples. However, not all genes were equally dysregulated and 25 genes were not upregulated at all. Genes located on other chromosomes were also significantly dysregulated. Functional class scoring and gene set enrichment analyses of 473 genes, differentially expressed between trisomic and non-trisomic hearts, revealed downregulation of genes encoding mitochondrial enzymes and upregulation of genes encoding extracellular matrix proteins. There were no significant differences between trisomic fetuses with and without heart defects. Conclusion We conclude that dosage-dependent upregulation of Hsa21 genes causes dysregulation of the genes responsible for mitochondrial function and for the extracellular matrix organization in the fetal heart of trisomic subjects. These alterations might be harbingers of the heart defects associated with Hsa21 trisomy, which could be based on elusive mechanisms involving genetic variability, environmental factors and/or stochastic events.

  9. The effect of enamel matrix derivative (Emdogain®) on gene expression profiles of human primary alveolar bone cells.

    Science.gov (United States)

    Yan, X Z; Rathe, F; Gilissen, C; van der Zande, M; Veltman, J; Junker, R; Yang, F; Jansen, J A; Walboomers, X F

    2014-06-01

    Emdogain® is frequently used in regenerative periodontal treatment. Understanding its effect on gene expression of bone cells would enable new products and pathways promoting bone formation to be established. The aim of the study was to analyse the effect of Emdogain® on expression profiles of human-derived bone cells with the help of the micro-array, and subsequent validation. Bone was harvested from non-smoking patients during dental implant surgery. After outgrowth, cells were cultured until subconfluence, treated for 24 h with either Emdogain® (100 µg/ml) or control medium, and subsequently RNA was isolated and micro-array was performed. The most important genes demonstrated by micro-array data were confirmed by qPCR and ELISA tests. Emdogain tipped the balance between genes expressed for bone formation and bone resorption towards a more anabolic effect, by interaction of the PGE2 pathway and inhibition of IL-7 production. In addition the results of the present study indicate that Emdogain possibly has an effect on gene expression for extracellular matrix formation of human bone cells, in particular on bone matrix formation and on proliferation and differentiation. With the micro-array and the subsequent validation, the genes possibly involved in Emdogain action on bone cells were identified. These results can contribute to establishing new products and pathways promoting bone formation.

  10. Cultivation of corneal endothelial cells on a pericellular matrix prepared from human decidua-derived mesenchymal cells.

    Science.gov (United States)

    Numata, Ryohei; Okumura, Naoki; Nakahara, Makiko; Ueno, Morio; Kinoshita, Shigeru; Kanematsu, Daisuke; Kanemura, Yonehiro; Sasai, Yoshiki; Koizumi, Noriko

    2014-01-01

    The barrier and pump functions of the corneal endothelium are essential for the maintenance of corneal transparency. Although corneal transplantation is the only current therapy for treating corneal endothelial dysfunction, the potential of tissue-engineering techniques to provide highly efficient and less invasive therapy in comparison to corneal transplantation has been highly anticipated. However, culturing human corneal endothelial cells (HCECs) is technically difficult, and there is no established culture protocol. The aim of this study was to investigate the feasibility of using a pericellular matrix prepared from human decidua-derived mesenchymal cells (PCM-DM) as an animal-free substrate for HCEC culture for future clinical applications. PCM-DM enhanced the adhesion of monkey CECs (MCECs) via integrin, promoted cell proliferation, and suppressed apoptosis. The HCECs cultured on the PCM-DM showed a hexagonal morphology and a staining profile characteristic of Na⁺/K⁺-ATPase and ZO-1 at the plasma membrane in vivo, whereas the control HCECs showed a fibroblastic phenotype. The cell density of the cultured HCECs on the PCM-DM was significantly higher than that of the control cells. These results indicate that PCM-DM provides a feasible xeno-free matrix substrate and that it offers a viable in vitro expansion protocol for HCECs while maintaining cellular functions for use as a subsequent clinical intervention for tissue-engineered based therapy of corneal endothelial dysfunction.

  11. Phase contrast imaging X-ray computed tomography: quantitative characterization of human patellar cartilage matrix with topological and geometrical features

    Science.gov (United States)

    Nagarajan, Mahesh B.; Coan, Paola; Huber, Markus B.; Diemoz, Paul C.; Wismüller, Axel

    2014-03-01

    Current assessment of cartilage is primarily based on identification of indirect markers such as joint space narrowing and increased subchondral bone density on x-ray images. In this context, phase contrast CT imaging (PCI-CT) has recently emerged as a novel imaging technique that allows a direct examination of chondrocyte patterns and their correlation to osteoarthritis through visualization of cartilage soft tissue. This study investigates the use of topological and geometrical approaches for characterizing chondrocyte patterns in the radial zone of the knee cartilage matrix in the presence and absence of osteoarthritic damage. For this purpose, topological features derived from Minkowski Functionals and geometric features derived from the Scaling Index Method (SIM) were extracted from 842 regions of interest (ROI) annotated on PCI-CT images of healthy and osteoarthritic specimens of human patellar cartilage. The extracted features were then used in a machine learning task involving support vector regression to classify ROIs as healthy or osteoarthritic. Classification performance was evaluated using the area under the receiver operating characteristic (ROC) curve (AUC). The best classification performance was observed with high-dimensional geometrical feature vectors derived from SIM (0.95 ± 0.06) which outperformed all Minkowski Functionals (p analysis of chondrocyte patterns in human patellar cartilage matrix involving SIM-derived geometrical features can distinguish between healthy and osteoarthritic tissue with high accuracy.

  12. Macrophage Inflammatory Protein-1alpha mediates Matrix Metalloproteinase-9 enhancement in human adherent monocytes fed with malarial pigment

    Institute of Scientific and Technical Information of China (English)

    Giuliana Giribaldi; Elena Valente; Amina Khadjavi; Manuela Polimeni; Mauro Prato

    2011-01-01

    Objective:To investigate the role of macrophage inflammatory protein-1alpha (MIP-1alpha) in the detrimental enhancement of matrix metalloproteinase-9 (MMP-9)expression, release and activity induced by phagocytosis of malarial pigment (haemozoin,HZ) in human monocytes. Methods: Human adherent monocytes were unfed/fed with nativeHZ for 2 h. After 24 hours, MIP-1alpha production was evaluated by ELISA in cell supernatants. Alternatively,HZ-unfed/fed monocytes were treated in presence/absence of anti-humanMIP-1alpha blocking antibodies or recombinant humanMIP-1alpha for15 h (RNA studies) or 24 h (protein studies); therefore,MMP-9mRNA expression was evaluated in cell lysates by Real TimeRT-PCR, whereas proMMP-9and activeMMP-9protein release were measured in cell supernatants by Western blotting and gelatin zymography.Results: Phagocytosis ofHZ by human monocytes increased production ofMIP-1alpha, mRNA expression ofMMP-9and protein release of proMMP-9 and activeMMP-9. All theHZ-enhancing effects onMMP-9 were abrogated by anti-humanMIP-1alpha blocking antibodies and mimicked by recombinant humanMIP-1alpha.Conclusions:The present work suggests a role for MIP-1alpha in theHZ-dependent enhancement ofMMP-9 expression, release and activity observed in human monocytes, highlighting new detrimental effects ofHZ-triggered proinflammatory response by phagocytic cells in falciparum malaria.

  13. A flexible matrix-based human exposure assessment framework suitable for LCA and CAA

    DEFF Research Database (Denmark)

    Jolliet, Olivier; Ernstoff, Alexi; Huang, Lei

    2016-01-01

    of near-and far-field pathways and helps to understand the contribution of individual pathways to overall human exposure in various product application contexts. When combined with toxicity information this approach is a resourceful way to inform LCA and CAA and minimize human exposure to toxic chemicals......Humans can be exposed to chemicals via near-field exposure pathways (e.g. through consumer product use) and far-field exposure pathways (e.g. through environmental emissions along product life cycles). Pathways are often complex where chemicals can transfer directly from products to humans during...... use or exchange between near-and far-field compartments until sub -fractions reach humans via inhalation, ingestion or dermal uptake. Currently, however, multimedia exposure models mainly focus on far-field exposure pathways. Metrics and modeling approaches used in far-field, emission-based models...

  14. The inhibitory effect of polyvinylphosphonic acid on functional matrix metalloproteinase activities in human demineralized dentin.

    Science.gov (United States)

    Tezvergil-Mutluay, Arzu; Agee, Kelli A; Hoshika, Tomohiro; Tay, Franklin R; Pashley, David H

    2010-10-01

    This study has examined the use of polyvinylphosphonic acid (PVPA) as a potential matrix metalloproteinase (MMP) inhibitor and how brief cross-linking of demineralized dentin matrix that did not affect its mechanical properties enhanced the anti-MMP activity of PVPA. The anti-MMP potential of five PVPA concentrations (100-3000 microgml(-1)) was initially screened using a rhMMP-9 colorimetic assay. Demineralized dentin beams were treated with the same five concentrations of PVPA to collagen and then aged for 30 days in a calcium- and zinc-containing medium. The changes in modulus of elasticity, loss of dry mass and dissolution of collagen peptides were measured via three-point bending, precision weighing and hydroxyproline assay, respectively. All tested PVPA concentrations were highly effective (P<0.05) in inhibiting MMP-9. Ageing in the incubation medium did not significantly alter the modulus of elasticity of the five PVPA treatment groups. Conversely, aged dentin beams from the control group exhibited a significant decline in their modulus of elasticity (P<0.05) over time. Mass loss from the dentin beams and the corresponding increase in hydroxyproline in the medium in the five PVPA treatment groups were significantly lower than for the control (P<0.05). PVPA is a potent inhibitor of endogenous MMP activities in demineralized dentin. It may be used as an alternative to chlorhexidine to prevent collagen degradation within hybrid layers to extend the longevity of resin-dentin bonds.

  15. Adsorbing/dissolving Lyoprotectant Matrix Technology for Non-cryogenic Storage of Archival Human Sera.

    Science.gov (United States)

    Solivio, Morwena J; Less, Rebekah; Rynes, Mathew L; Kramer, Marcus; Aksan, Alptekin

    2016-04-12

    Despite abundant research conducted on cancer biomarker discovery and validation, to date, less than two-dozen biomarkers have been approved by the FDA for clinical use. One main reason is attributed to inadvertent use of low quality biospecimens in biomarker research. Most proteinaceous biomarkers are extremely susceptible to pre-analytical factors such as collection, processing, and storage. For example, cryogenic storage imposes very harsh chemical, physical, and mechanical stresses on biospecimens, significantly compromising sample quality. In this communication, we report the development of an electrospun lyoprotectant matrix and isothermal vitrification methodology for non-cryogenic stabilization and storage of liquid biospecimens. The lyoprotectant matrix was mainly composed of trehalose and dextran (and various low concentration excipients targeting different mechanisms of damage), and it was engineered to minimize heterogeneity during vitrification. The technology was validated using five biomarkers; LDH, CRP, PSA, MMP-7, and C3a. Complete recovery of LDH, CRP, and PSA levels was achieved post-rehydration while more than 90% recovery was accomplished for MMP-7 and C3a, showing promise for isothermal vitrification as a safe, efficient, and low-cost alternative to cryogenic storage.

  16. Peroxidase Enzymes Regulate Collagen Biosynthesis and Matrix Mineralization by Cultured Human Osteoblasts.

    Science.gov (United States)

    DeNichilo, Mark O; Shoubridge, Alexandra J; Panagopoulos, Vasilios; Liapis, Vasilios; Zysk, Aneta; Zinonos, Irene; Hay, Shelley; Atkins, Gerald J; Findlay, David M; Evdokiou, Andreas

    2016-03-01

    The early recruitment of inflammatory cells to sites of bone fracture and trauma is a critical determinant in successful fracture healing. Released by infiltrating inflammatory cells, myeloperoxidase (MPO) and eosinophil peroxidase (EPO) are heme-containing enzymes, whose functional involvement in bone repair has mainly been studied in the context of providing a mechanism for oxidative defense against invading microorganisms. We report here novel findings that show peroxidase enzymes have the capacity to stimulate osteoblastic cells to secrete collagen I protein and generate a mineralized extracellular matrix in vitro. Mechanistic studies conducted using cultured osteoblasts show that peroxidase enzymes stimulate collagen biosynthesis at a post-translational level in a prolyl hydroxylase-dependent manner, which does not require ascorbic acid. Our studies demonstrate that osteoblasts rapidly bind and internalize both MPO and EPO, and the catalytic activity of these peroxidase enzymes is essential to support collagen I biosynthesis and subsequent release of collagen by osteoblasts. We show that EPO is capable of regulating osteogenic gene expression and matrix mineralization in culture, suggesting that peroxidase enzymes may play an important role not only in normal bone repair, but also in the progression of pathological states where infiltrating inflammatory cells are known to deposit peroxidases.

  17. Enamel matrix derivative promote primary human pulp cell differentiation and mineralization

    National Research Council Canada - National Science Library

    Riksen, Elisabeth Aurstad; Landin, Maria A; Reppe, Sjur; Nakamura, Yukio; Lyngstadaas, Ståle Petter; Reseland, Janne E

    2014-01-01

    ...; however the molecular mechanisms involved are unclear. The effect of EMD (5-50 μg/mL) on primary human pulp cells were compared to untreated cells and cells incubated with 10⁻⁸ M dexamethasone (DEX...

  18. Nanofibers coated on acellular tissue-engineered bovine pericardium supports differentiation of mesenchymal stem cells into endothelial cells for tissue engineering.

    Science.gov (United States)

    Mathapati, Santosh; Bishi, Dillip Kumar; Venugopal, Jayarama Reddy; Cherian, Kotturathu Mammen; Guhathakurta, Soma; Ramakrishna, Seeram; Verma, Rama Shanker

    2014-04-01

    This study aimed to develop biodegradable, polymer-based nanofibers coated on acellular tissue-engineered bovine pericardium (ATEBP) for cell interfaces, enabling more exquisite functionality, such as mesenchymal stem cell (MSC) adhesion, proliferation and differentiation into endothelial cells for tissue engineering. ATEBP coated with nanofibers of poly(L-lactic acid)-co-poly(ε-caprolactone) (PLACL) and a blend of PLACL and gelatin were analyzed for human bone marrow-derived MSC adhesion, proliferation and differentiation into endothelial cells. The cell culture-based approach showed an increase in human bone marrow-derived MSC adhesion, proliferation and differentiation into endothelial cells on ATEBP coated with PLACL/gelatin nanofibers compared with ATEBP and PLACL nanofibers coated on ATEBP. ATEBP coated with PLACL/gelatin nanofibrous scaffolds, along with human bone marrow-derived MSCs differentiated into endothelial cells, might improve the scaffolds' functionality for tissue engineering.

  19. Categorical dimensions of human odor descriptor space revealed by non-negative matrix factorization

    Energy Technology Data Exchange (ETDEWEB)

    Chennubhotla, Chakra [University of Pittsburgh School of Medicine, Pittsburgh PA; Castro, Jason [Bates College

    2013-01-01

    In contrast to most other sensory modalities, the basic perceptual dimensions of olfaction remain un- clear. Here, we use non-negative matrix factorization (NMF) - a dimensionality reduction technique - to uncover structure in a panel of odor profiles, with each odor defined as a point in multi-dimensional descriptor space. The properties of NMF are favorable for the analysis of such lexical and perceptual data, and lead to a high-dimensional account of odor space. We further provide evidence that odor di- mensions apply categorically. That is, odor space is not occupied homogenously, but rather in a discrete and intrinsically clustered manner. We discuss the potential implications of these results for the neural coding of odors, as well as for developing classifiers on larger datasets that may be useful for predicting perceptual qualities from chemical structures.

  20. TCDD induces dermal accumulation of keratinocyte-derived matrix metalloproteinase-10 in an organotypic model of human skin.

    Science.gov (United States)

    De Abrew, K Nadira; Thomas-Virnig, Christina L; Rasmussen, Cathy A; Bolterstein, Elyse A; Schlosser, Sandy J; Allen-Hoffmann, B Lynn

    2014-05-01

    The epidermis of skin is the first line of defense against the environment. A three dimensional model of human skin was used to investigate tissue-specific phenotypes induced by the environmental contaminant, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Continuous treatment of organotypic cultures of human keratinocytes with TCDD resulted in intracellular spaces between keratinocytes of the basal and immediately suprabasal layers as well as thinning of the basement membrane, in addition to the previously reported hyperkeratinization. These tissue remodeling events were preceded temporally by changes in expression of the extracellular matrix degrading enzyme, matrix metalloproteinase-10 (MMP-10). In organotypic cultures MMP-10 mRNA and protein were highly induced following TCDD treatment. Q-PCR and immunoblot results from TCDD-treated monolayer cultures, as well as indirect immunofluorescence and immunoblot analysis of TCDD-treated organotypic cultures, showed that MMP-10 was specifically contributed by the epidermal keratinocytes but not the dermal fibroblasts. Keratinocyte-derived MMP-10 protein accumulated over time in the dermal compartment of organotypic cultures. TCDD-induced epidermal phenotypes in organotypic cultures were attenuated by the keratinocyte-specific expression of tissue inhibitor of metalloproteinase-1, a known inhibitor of MMP-10. These studies suggest that MMP-10 and possibly other MMP-10-activated MMPs are responsible for the phenotypes exhibited in the basement membrane, the basal keratinocyte layer, and the cornified layer of TCDD-treated organotypic cultures. Our studies reveal a novel mechanism by which the epithelial-stromal microenvironment is altered in a tissue-specific manner thereby inducing structural and functional pathology in the interfollicular epidermis of human skin.

  1. The interaction between epidermal growth factor (EGF) and matrix metalloproteinase induces the development of sweat glands in human fetal skin

    Institute of Scientific and Technical Information of China (English)

    Li Jianfu; Fu Xiaobing; Sheng Zhiyong

    2001-01-01

    Objective:The development of sweat glands is a very complicated biological process involving many factors. In this study, we explore the inter-relationship between epidermal growth factor (EGF),matrix metalloproteinases (MMP-2,MMP-7) and development of sweat glands in human embryos. Furthermore, we hope to elucidate the mechanism(s) underlying the induction of epidermal stem cells into sweat gland cells. Methods:Skin biospies of human embryos obtained from spontaneous abortions at different gestational ages from 11 to 31 weeks were used in this study. The dynamical expression of EGF, MMP-2, MMP-7 and keratin-7 (K7) in developing sweat gland cells or extracellular stroma surrounding the sweat gland cells were examined with S-P immunohistochemical methods.The localization of the cellular sources of MMP-2 and MMP 7 was examined with in situ hybridization. Results:At 14-20 wk of gestation, a gradual increase in EGF immunoreactivity was observed not only in developing sweat gland buds but also in extracellular stroma surrounding the buds,and the expression intensity peaked at 20-22 wk of gesta- tional age. All mRNA-positive buds or cells in developing sweat glands contained corresponding immunoreactive proteins. Positive immunostaining for K7 appeared in early sweat gland buds at 14-16wk of gestation, and from then on, K7 was concentrated in developing sweat gland cords or cells. Conclusions: The morphogenesis of sweat gland in human fetal skin begins at 14-16wk of gestational age, and essentially completes by 24wk. There is a close relationship among EGF,extracellular matrix remodeling and morphogenesis of sweat glands, and EGF is one of the inducers in the development and maturity of sweat gland buds or cells.

  2. Seeding for sirtuins: microseed matrix seeding to obtain crystals of human Sirt3 and Sirt2 suitable for soaking

    Energy Technology Data Exchange (ETDEWEB)

    Rumpf, Tobias [Albert-Ludwigs-University Freiburg, Albertstrasse 25, 79104 Freiburg, Baden-Württemberg (Germany); Gerhardt, Stefan; Einsle, Oliver, E-mail: einsle@biochemie.uni-freiburg.de [Albert-Ludwigs-University Freiburg, Albertstrasse 21, 79104 Freiburg, Baden-Württemberg (Germany); Jung, Manfred, E-mail: einsle@biochemie.uni-freiburg.de [Albert-Ludwigs-University Freiburg, Albertstrasse 25, 79104 Freiburg, Baden-Württemberg (Germany)

    2015-11-18

    In the present study, microseed matrix seeding was successfully applied to obtain a large number of crystals of the human sirtuin isotypes Sirt2 and Sirt3. These crystals appeared predictably in diverse crystallization conditions, diffracted to a higher resolution than reported in the literature and were subsequently used to study the protein–ligand interactions of two indole inhibitors. Sirtuins constitute a family of NAD{sup +}-dependent enzymes that catalyse the cleavage of various acyl groups from the ∊-amino group of lysines. They regulate a series of cellular processes and their misregulation has been implicated in various diseases, making sirtuins attractive drug targets. To date, only a few sirtuin modulators have been reported that are suitable for cellular research and their development has been hampered by a lack of structural information. In this work, microseed matrix seeding (MMS) was used to obtain crystals of human Sirt3 in its apo form and of human Sirt2 in complex with ADP ribose (ADPR). Crystal formation using MMS was predictable, less error-prone and yielded a higher number of crystals per drop than using conventional crystallization screening methods. The crystals were used to solve the crystal structures of apo Sirt3 and of Sirt2 in complex with ADPR at an improved resolution, as well as the crystal structures of Sirt2 in complex with ADPR and the indoles EX527 and CHIC35. These Sirt2–ADPR–indole complexes unexpectedly contain two indole molecules and provide novel insights into selective Sirt2 inhibition. The MMS approach for Sirt2 and Sirt3 may be used as the basis for structure-based optimization of Sirt2/3 inhibitors in the future.

  3. Comparison of structural, architectural and mechanical aspects of cellular and acellular bone in two teleost fish.

    Science.gov (United States)

    Cohen, Liat; Dean, Mason; Shipov, Anna; Atkins, Ayelet; Monsonego-Ornan, Efrat; Shahar, Ron

    2012-06-01

    The histological diversity of the skeletal tissues of fishes is impressive compared with that of other vertebrate groups, yet our understanding of the functional consequences of this diversity is limited. In particular, although it has been known since the mid-1800s that a large number of fish species possess acellular bones, the mechanical advantages and consequences of this structural characteristic - and therefore the nature of the evolution of this feature - remain unclear. Although several studies have examined the material properties of fish bone, these have used a variety of techniques and there have been no direct contrasts of acellular and cellular bone. We report on a comparison of the structural and mechanical properties of the ribs and opercula between two freshwater fish - the common carp Cyprinus carpio (a fish with cellular bone) and the tilapia Oreochromis aureus (a fish with acellular bone). We used light microscopy to show that the bones in both fish species exhibit poor blood supply and possess discrete tissue zones, with visible layering suggesting differences in the underlying collagen architecture. We performed identical micromechanical testing protocols on samples of the two bone types to determine the mechanical properties of the bone material of opercula and ribs. Our data support the consensus of literature values, indicating that Young's moduli of cellular and acellular bones are in the same range, and lower than Young's moduli of the bones of mammals and birds. Despite these similarities in mechanical properties between the bone tissues of the fish species tested here, cellular bone had significantly lower mineral content than acellular bone; furthermore, the percentage ash content and bone mineral density values (derived from micro-CT scans) show that the bone of these fishes is less mineralized than amniote bone. Although we cannot generalize from our data to the numerous remaining teleost species, the results presented here suggest

  4. Histochemical evidence of osteoclastic degradation of extracellular matrix in osteolytic metastasis originating from human lung small carcinoma (SBC-5) cells.

    Science.gov (United States)

    Li, Minqi; Amizuka, Norio; Takeuchi, Kiichi; Freitas, Paulo H L; Kawano, Yoshiro; Hoshino, Masaaki; Oda, Kimimitsu; Nozawa-Inoue, Kayoko; Maeda, Takeyasu

    2006-02-01

    The aim of this study was to assess the dynamics of osteoclast migration and the degradation of unmineralized extracellular matrix in an osteolytic metastasis by examining a well-standardized lung cancer metastasis model of nude mice. SBC-5 human lung small carcinoma cells were injected into the left cardiac ventricle of 6-week-old BALB/c nu/nu mice under anesthesia. At 25-30 days after injection, the animals were sacrificed and their femora and/or tibiae were removed for histochemical analyses. Metastatic lesions were shown to occupy a considerable area extending from the metaphyses to the bone marrow region. Tartrate resistant acid phosphatase (TRAPase)-positive osteoclasts were found in association with an alkaline phosphatase (ALPase)-positive osteoblastic layer lining the bone surface, but could also be localized in the ALPase-negative stromal tissues that border the tumor nodules. These stromal tissues were markedly positive for osteopontin, and contained a significant number of TRAPase-positive osteoclasts expressing immunoreactivity for CD44. We thus speculated that, mediating its affinity for CD44, osteopontin may serve to facilitate osteoclastic migration after their formation associated with ALPase-positive osteoblasts. We next examined the localization of cathepsin K and matrix metallo-proteinase-9 (MMP-9) in osteoclasts. Osteoclasts adjacent to the bone surfaces were positive for both proteins, whereas those in the stromal tissues in the tumor nests showed only MMP-9 immunoreactivity. Immunoelectron microscopy disclosed the presence of MMP-9 in the Golgi apparatus and in vesicular structures at the baso-lateral cytoplasmic region of the osteoclasts found in the stromal tissue. MMP-9-positive vesicular structures also contained fragmented extracellular materials. Thus, osteoclasts appear to either select an optimized function, namely secreting proteolytic enzymes from ruffled borders during bone resorption, or recognize the surrounding extracellular

  5. Ornithine decarboxylase, mitogen-activated protein kinase and matrix metalloproteinase-2 expressions in human colon tumors

    Institute of Scientific and Technical Information of China (English)

    Takahiro Nemoto; Shunichiro Kubota; Hideyuki Ishida; Nobuo Murata; Daijo Hashimoto

    2005-01-01

    AIM: To investigate the expressions of omithine decarboxylase (ODC), MMP-2, and Erk, and their relationship in human colon tumors.METHODS: ODC activity, MMP-2 expression, and mitogenactivated protein (MAP) kinase activity (Erk phosphorylation) were determined in 58 surgically removed human colon tumors and their adjacent normal tissues, using [1-14C]-ornithine as a substrate, ELISA assay, and Western blotting, respectively.RESULTS: ODC activity, MMP-2 expression, and Erk phosphorylation were significantly elevated in colon tumors, compared to those in adjacent normal tissues. A significant correlation was observed between ODC activities and MMP-2 levels.CONCLUSION: This is the first report showing a significant correlation between ODC activities and MMP-2 levels in human colon tumors. As MMP-2 is involved in cancer invasion and metastasis, and colon cancer overexpresses ODC, suppression of ODC expression may be a rational approach to treat colon cancer which overexpresses ODC.

  6. GH/IGF-I axis and matrix adaptation of the musculotendinous tissue to exercise in humans

    DEFF Research Database (Denmark)

    Heinemeier, K M; Mackey, Abigail; Doessing, S

    2012-01-01

    Exercise is not only associated with adaptive responses within skeletal muscle fibers but also with induction of collagen synthesis both in muscle and adjacent connective tissue. Additionally, exercise and training leads to activation of the systemic growth hormone/insulin-like growth factor I axis...... (GH/IGF-I), as well as increased local IGF-I expression. Studies in humans with pathologically high levels of GH/IGF-I, and in healthy humans who receive either weeks of GH administration or acute injection of IGF-I into connective tissue, demonstrate increased expression and synthesis of collagen...

  7. MicroRNA-143 Regulates Human Osteosarcoma Metastasis by Regulating Matrix Metalloprotease-13 Expression

    OpenAIRE

    Osaki, Mitsuhiko; Takeshita, Fumitaka; Sugimoto, Yui; Kosaka, Nobuyoshi; Yamamoto, Yusuke; Yoshioka, Yusuke; Kobayashi,Eisuke; Yamada, Tesshi; Kawai, Akira; Inoue, Toshiaki; Ito, Hisao; Oshimura, Mitsuo; Ochiya, Takahiro

    2011-01-01

    Pulmonary metastases are the main cause of death in patients with osteosarcoma, however, the molecular mechanisms of metastasis are not well understood. To detect lung metastasis-related microRNA (miRNA) in human osteosarcoma, we compared parental (HOS) and its subclone (143B) human osteosarcoma cell lines showing lung metastasis in a mouse model. miR-143 was the most downregulated miRNA (P < 0.01), and transfection of miR-143 into 143B significantly decreased its invasiveness, but not cell p...

  8. Tantalum coating on TiO{sub 2} nanotubes induces superior rate of matrix mineralization and osteofunctionality in human osteoblasts

    Energy Technology Data Exchange (ETDEWEB)

    Frandsen, Christine J.; Brammer, Karla S. [Materials Science and Engineering, University of California at San Diego, La Jolla, CA 92093 (United States); Noh, Kunbae [Corporate Research Institute, Cheil Industries, Inc., Gocheon-Dong, Uiwang-Si, Gyeonggi-Do, 437-711 (Korea, Republic of); Johnston, Gary [Materials Science and Engineering, University of California at San Diego, La Jolla, CA 92093 (United States); Jin, Sungho, E-mail: jin@ucsd.edu [Materials Science and Engineering, University of California at San Diego, La Jolla, CA 92093 (United States); Mechanical and Aerospace Engineering, University of California at San Diego, La Jolla, CA 92093 (United States)

    2014-04-01

    Nanostructured surface geometries have been the focus of a multitude of recent biomaterial research, and exciting findings have been published. However, only a few publications have directly compared nanostructures of various surface chemistries. The work herein directly compares the response of human osteoblast cells to surfaces of identical nanotube geometries with two well-known orthopedic biomaterials: titanium oxide (TiO{sub 2}) and tantalum (Ta). The results reveal that the Ta surface chemistry on the nanotube architecture enhances alkaline phosphatase activity, and promotes a ∼ 30% faster rate of matrix mineralization and bone-nodule formation when compared to results on bare TiO{sub 2} nanotubes. This study implies that unique combinations of surface chemistry and nanostructure may influence cell behavior due to distinctive physico-chemical properties. These findings are of paramount importance to the orthopedics field for understanding cell behavior in response to subtle alterations in nanostructure and surface chemistry, and will enable further insight into the complex manipulation of biomaterial surfaces. With increased focus in the field of orthopedic materials research on nanostructured surfaces, this study emphasizes the need for careful and systematic review of variations in surface chemistry in concurrence with nanotopographical changes. - Highlights: • A TiO{sub 2} nanotube surface structure was coated with tantalum. • Osteoblast cell response was compared between the tantalum coated and as-formed TiO{sub 2} nanotube surface. • We observed superior rates of bone matrix mineralization and osteoblast maturation on the tantalum coated nanotube surface.

  9. Skip Regulates TGF-β1-Induced Extracellular Matrix Degrading Proteases Expression in Human PC-3 Prostate Cancer Cells

    Directory of Open Access Journals (Sweden)

    Victor Villar

    2013-01-01

    Full Text Available Purpose. To determine whether Ski-interacting protein (SKIP regulates TGF-β1-stimulated expression of urokinase-type plasminogen activator (uPA, matrix metalloproteinase-9 (MMP-9, and uPA Inhibitor (PAI-1 in the androgen-independent human prostate cancer cell model. Materials and Methods. PC-3 prostate cancer cell line was used. The role of SKIP was evaluated using synthetic small interference RNA (siRNA compounds. The expression of uPA, MMP-9, and PAI-1 was evaluated by zymography assays, RT-PCR, and promoter transactivation analysis. Results. In PC-3 cells TGF-β1 treatment stimulated uPA, PAI-1, and MMP-9 expressions. The knockdown of SKIP in PC-3 cells enhanced the basal level of uPA, and TGF-β1 treatment inhibited uPA production. Both PAI-1 and MMP-9 production levels were increased in response to TGF-β1. The ectopic expression of SKIP inhibited both TGF-β1-induced uPA and MMP-9 promoter transactivation, while PAI-1 promoter response to the factor was unaffected. Conclusions. SKIP regulates the expression of uPA, PAI-1, and MMP-9 stimulated by TGF-β1 in PC-3 cells. Thus, SKIP is implicated in the regulation of extracellular matrix degradation and can therefore be suggested as a novel therapeutic target in prostate cancer treatment.

  10. Rodent neonatal germinal matrix hemorrhage mimics the human brain injury, neurological consequences, and post-hemorrhagic hydrocephalus.

    Science.gov (United States)

    Lekic, Tim; Manaenko, Anatol; Rolland, William; Krafft, Paul R; Peters, Regina; Hartman, Richard E; Altay, Orhan; Tang, Jiping; Zhang, John H

    2012-07-01

    Germinal matrix hemorrhage (GMH) is the most common neurological disease of premature newborns. GMH causes neurological sequelae such as cerebral palsy, post-hemorrhagic hydrocephalus, and mental retardation. Despite this, there is no standardized animal model of spontaneous GMH using newborn rats to depict the condition. We asked whether stereotactic injection of collagenase type VII (0.3 U) into the ganglionic eminence of neonatal rats would reproduce the acute brain injury, gliosis, hydrocephalus, periventricular leukomalacia, and attendant neurological consequences found in humans. To test this hypothesis, we used our neonatal rat model of collagenase-induced GMH in P7 pups, and found that the levels of free-radical adducts (nitrotyrosine and 4-hyroxynonenal), proliferation (mammalian target of rapamycin), inflammation (COX-2), blood components (hemoglobin and thrombin), and gliosis (vitronectin and GFAP) were higher in the forebrain of GMH pups, than in controls. Neurobehavioral testing showed that pups with GMH had developmental delay, and the juvenile animals had significant cognitive and motor disability, suggesting clinical relevance of the model. There was also evidence of white-matter reduction, ventricular dilation, and brain atrophy in the GMH animals. This study highlights an instructive animal model of the neurological consequences after germinal matrix hemorrhage, with evidence of brain injuries that can be used to evaluate strategies in the prevention and treatment of post-hemorrhagic complications.

  11. Matrix-degrading and pro-inflammatory changes in human vascular endothelial cells exposed to cigarette smoke condensate.

    Science.gov (United States)

    Nordskog, Brian K; Blixt, Allison D; Morgan, Walter T; Fields, Wanda R; Hellmann, Gary M

    2003-01-01

    Cigarette smoking has been associated with an increase in the severity and prevalence of atherosclerosis in the abdominal aorta. To begin our investigation of this finding, we used an integrated approach combining gene expression profiling, protein analysis, cytokine measurements, and cytotoxicity determinations to examine molecular responses of cultured human aortic and coronary endothelial cells exposed to cigarette smoke condensate (CSC) and nicotine. Exposure of endothelial cells to CSC (30 and 60 microg/mL TPM) for 24 h resulted in minimal cytotoxicity, and the upregulation of genes involved in matrix degradation (MMP-1, MMP-8, and MMP-9), xenobiotic metabolism (HO-1 and CYP1A2), and downregulation of genes involved in cell cycle regulation (including TOP2A, CCNB1, CCNA, CDKN3). Exposure of cells to a high physiological concentration of nicotine resulted in few differentially expressed genes. Immunoblot analysis of proteins selected from genes shown to be differentially regulated by microarray analysis revealed similar responses. Finally, a number of inflammatory cytokines measured in culture media were elevated in response to CSC. Together, these results describe a complex proinflammatory response, possibly mediating the recruitment of leukocytes through cytokine signaling. Additionally, fibrous cap destabilization may be facilitated by matrix metalloproteinase upregulation.

  12. Role of Nuclear Matrix in Estrogen Regulated Gene Expression in Human Breast Cancer Cells

    Science.gov (United States)

    1998-08-01

    form of endocrine manipulative therapy, e.g., antiestrogen therapy. However, most human breast cancers originate as hormonally dependent tumors as...development. 49 "Proprietary Data - Distribution to Government Agencies Only" ACKNOWLEDGMENTS Pierre Chambon (Institut de Genetique et de Biologie

  13. The Effects of Water Matrix on Decay of Human Fecal Molecular Markers and Campylobacter spp.

    Science.gov (United States)

    Although molecular source tracking for human fecal contamination is used on a wide range of sample types, little is known about comparative decay of proposed molecular markers under different conditions, or correlation with pathogen decay. Our purpose was to measure correlations ...

  14. TEMPERATURE-DEPENDENCE OF WATER TRANSPORT INTO THE MINERALIZED MATRIX OF FREEZE-DRIED HUMAN DENTIN

    NARCIS (Netherlands)

    VANDERGRAAF, ER; TENBOSCH, JJ

    1991-01-01

    Ten dentine sections cut perpendicular to the dentinal tubules from human mature non-carious third molars, were freeze-dried and then rehydrated by immersion in water at four temperatures, 10, 25, 40 and 70-degrees-C. The uptake of water by the sections was assessed as a function of rehydration time

  15. The Effects of Water Matrix on Decay of Human Fecal Molecular Markers and Campylobacter spp.

    Science.gov (United States)

    Although molecular source tracking for human fecal contamination is used on a wide range of sample types, little is known about comparative decay of proposed molecular markers under different conditions, or correlation with pathogen decay. Our purpose was to measure correlations ...

  16. Optical diagnosis of dengue virus infected human blood using Mueller matrix polarimetry

    Science.gov (United States)

    Anwar, Shahzad; Firdous, Shamaraz

    2016-08-01

    Currently dengue fever diagnosis methods include capture ELISAs, immunofluorescence tests, and hemagglutination assays. In this study optical diagnosis of dengue virus infection in the whole blood is presented utilizing Mueller matrix polarimetry. Mueller matrices of about 50 dengue viral infected and 25 non-dengue healthy blood samples were recorded utilizing light source from 500 to 700 nm with scanning step of 10 nm. Polar decomposition of the Mueller matrices for all the blood samples was performed that yielded polarization properties including depolarization, diattenuation, degree of polarization, retardance and optical activity, out of which, depolarization index clusters up the diseased and healthy in to different separate groups. The average depolarized light in the case of dengue infection in the whole blood at 500 nm is 18%, whereas for the healthy blood samples it is 13.5%. This suggests that depolarization index of polarized light at the wavelengths of 500, 510, 520, 530 and 540 nm, we find that in case of depolarization index values are higher for dengue viral infection as compared to normal samples. This technique can effectively be used for the characterization of the dengue virus infected at an early stage of disease.

  17. Changes in vascular extracellular matrix composition during decidual spiral arteriole remodeling in early human pregnancy.

    Science.gov (United States)

    Smith, Samantha D; Choudhury, Ruhul H; Matos, Patricia; Horn, James A; Lye, Stephen J; Dunk, Caroline E; Aplin, John D; Jones, Rebecca L; Harris, Lynda K

    2016-05-01

    Uterine spiral arteriole (SA) remodeling in early pregnancy involves a coordinated series of events including decidual immune cell recruitment, vascular cell disruption and loss, and colonization by placental-derived extravillous trophoblast (EVT). During this process, decidual SA are converted from narrow, muscular vessels into dilated channels lacking vasomotor control. We hypothesized that this extensive alteration in SA architecture must require significant reorganization and/or breakdown of the vascular extracellular matrix (ECM). First trimester decidua basalis (30 specimens) was immunostained to identify spiral arterioles undergoing trophoblast-independent and -dependent phases of remodeling. Serial sections were then immunostained for a panel of ECM markers, to examine changes in vascular ECM during the remodeling process. The initial stages of SA remodeling were characterized by loss of laminin, elastin, fibrillin, collagen types III, IV and VI from the basement membrane, vascular media and/or adventitia, and surrounding decidual stromal cells. Loss of ECM correlated with disruption and disorganization of vascular smooth muscle cells, and the majority of changes occurred prior to extensive colonization of the vessel wall by EVT. The final stages of SA remodeling, characterized by the arrival of EVT, were associated with the increased mural deposition of fibronectin and fibrinoid. This study provides the first detailed analysis of the spatial and temporal loss of ECM from the walls of remodeling decidual SA in early pregnancy.

  18. Expression of matrix metalloproteinases 2 and 9 in human gastric cancer and superficial gastritis.

    Science.gov (United States)

    Sampieri, Clara Luz; de la Peña, Sol; Ochoa-Lara, Mariana; Zenteno-Cuevas, Roberto; León-Córdoba, Kenneth

    2010-03-28

    To assess expression of matrix metalloproteinases 2 (MMP2) and MMP9 in gastric cancer, superficial gastritis and normal mucosa, and to measure metalloproteinase activity. MMP2 and MMP9 mRNA expression was determined by quantitative real-time polymerase chain reaction. Normalization was carried out using three different factors. Proteins were analyzed by quantitative gelatin zymography (qGZ). 18S ribosomal RNA (18SRNA) was very highly expressed, while hypoxanthine ribosyltransferase-1 (HPRT-1) was moderately expressed. MMP2 was highly expressed, while MMP9 was not detected or lowly expressed in normal tissues, moderately or highly expressed in gastritis and highly expressed in cancer. Relative expression of 18SRNA and HPRT-1 showed no significant differences. Significant differences in MMP2 and MMP9 were found between cancer and normal tissue, but not between gastritis and normal tissue. Absolute quantification of MMP9 echoed this pattern, but differential expression of MMP2 proved conflictive. Analysis by qGZ indicated significant differences between cancer and normal tissue in MMP-2, total MMP-9, 250 and 110 kDa bands. MMP9 expression is enhanced in gastric cancer compared to normal mucosa; interpretation of differential expression of MMP2 is difficult to establish.

  19. Imaging-guided two-photon excitation-emission-matrix measurements of human skin tissues

    Science.gov (United States)

    Yu, Yingqiu; Lee, Anthony M. D.; Wang, Hequn; Tang, Shuo; Zhao, Jianhua; Lui, Harvey; Zeng, Haishan

    2012-07-01

    There are increased interests on using multiphoton imaging and spectroscopy for skin tissue characterization and diagnosis. However, most studies have been done with just a few excitation wavelengths. Our objective is to perform a systematic study of the two-photon fluorescence (TPF) properties of skin fluorophores, normal skin, and diseased skin tissues. A nonlinear excitation-emission-matrix (EEM) spectroscopy system with multiphoton imaging guidance was constructed. A tunable femtosecond laser was used to vary excitation wavelengths from 730 to 920 nm for EEM data acquisition. EEM measurements were performed on excised fresh normal skin tissues, seborrheic keratosis tissue samples, and skin fluorophores including: NADH, FAD, keratin, melanin, collagen, and elastin. We found that in the stratum corneum and upper epidermis of normal skin, the cells have large sizes and the TPF originates from keratin. In the lower epidermis, cells are smaller and TPF is dominated by NADH contributions. In the dermis, TPF is dominated by elastin components. The depth resolved EEM measurements also demonstrated that keratin structure has intruded into the middle sublayers of the epidermal part of the seborrheic keratosis lesion. These results suggest that the imaging guided TPF EEM spectroscopy provides useful information for the development of multiphoton clinical devices for skin disease diagnosis.

  20. Elevated expression of matrix metalloproteinase-3 in human osteosarcoma and its association with tumor metastasis.

    Science.gov (United States)

    Huang, Jie-Feng; Du, Wen-Xi; Chen, Jun-Jie

    2016-01-01

    Matrix metalloproteinase-3 (MMP-3) is one of the several MMPs that is associated with malignant tumors of breast, colon, cervix and lung, where its expression has been correlated with tumor invasion and metastasis. However, the role of MMP-3 in metastasis of osteosarcoma has not yet been explored. MMP-3 expression in 15 primary and metastatic osteosarcomas with case-matched adjacent non-tumor tissue was assessed by immunohistochemistry and quantitative RT-PCR. Further, MMP-3 mRNA and protein levels were also determined in osteoblast and osteosarcoma cell lines. Additionally, migration and invasion assays were performed in MMP-3 knockdown cells. MMP-3 was expressed in 86.6% (13/15) of the osteosarcoma patients and its expression was significantly higher in metastatic tumors as compared to the primary osteosarcoma tumor tissues. Furthermore, osteosarcoma cell lines showed higher MMP-3 expression as compared to osteoblast cell lines. siRNA-mediated MMP-3 knockdown in osteosarcoma cell lines significantly inhibited their migration and invasion properties. Our results demonstrated that MMP-3 expression is deregulated in osteosarcomas and this potentially contributes to metastasis and might be a promising marker for the prognosis and therapy of metastatic osteosarcoma.

  1. Adhesion of the human pathogen Sporothrix schenckii to several extracellular matrix proteins

    Directory of Open Access Journals (Sweden)

    O.C. Lima

    1999-05-01

    Full Text Available The pathogenic fungus Sporothrix schenckii is the causative agent of sporotrichosis. This subcutaneous mycosis may disseminate in immunocompromised individuals and also affect several internal organs and tissues, most commonly the bone, joints and lung. Since adhesion is the first step involved with the dissemination of pathogens in the host, we have studied the interaction between S. schenckii and several extracellular matrix (ECM proteins. The binding of two morphological phases of S. schenckii, yeast cells and conidia, to immobilized type II collagen, laminin, fibronectin, fibrinogen and thrombospondin was investigated. Poly (2-hydroxyethyl methacrylate (poly-HEMA was used as the negative control. Cell adhesion was assessed by ELISA with a rabbit anti-S. schenckii antiserum. The results indicate that both morphological phases of this fungus can bind significantly to type II collagen, fibronectin and laminin in comparison to the binding observed with BSA (used as blocking agent. The adhesion rate observed with the ECM proteins (type II collagen, fibronectin and laminin was statistically significant (P<0.05 when compared to the adhesion obtained with BSA. No significant binding of conidia was observed to either fibrinogen or thrombospondin, but yeast cells did bind to the fibrinogen. Our results indicate that S. schenckii can bind to fibronectin, laminin and type II collagen and also show differences in binding capacity according to the morphological form of the fungus.

  2. Expression of matrix metalloproteinases 2 and 9 in human gastric cancer and superficial gastritis

    Science.gov (United States)

    Sampieri, Clara Luz; de la Peña, Sol; Ochoa-Lara, Mariana; Zenteno-Cuevas, Roberto; León-Córdoba, Kenneth

    2010-01-01

    AIM: To assess expression of matrix metalloproteinases 2 (MMP2) and MMP9 in gastric cancer, superficial gastritis and normal mucosa, and to measure metalloproteinase activity. METHODS: MMP2 and MMP9 mRNA expression was determined by quantitative real-time polymerase chain reaction. Normalization was carried out using three different factors. Proteins were analyzed by quantitative gelatin zymography (qGZ). RESULTS: 18S ribosomal RNA (18SRNA) was very highly expressed, while hypoxanthine ribosyltransferase-1 (HPRT-1) was moderately expressed. MMP2 was highly expressed, while MMP9 was not detected or lowly expressed in normal tissues, moderately or highly expressed in gastritis and highly expressed in cancer. Relative expression of 18SRNA and HPRT-1 showed no significant differences. Significant differences in MMP2 and MMP9 were found between cancer and normal tissue, but not between gastritis and normal tissue. Absolute quantification of MMP9 echoed this pattern, but differential expression of MMP2 proved conflictive. Analysis by qGZ indicated significant differences between cancer and normal tissue in MMP-2, total MMP-9, 250 and 110 kDa bands. CONCLUSION: MMP9 expression is enhanced in gastric cancer compared to normal mucosa; interpretation of differential expression of MMP2 is difficult to establish. PMID:20333791

  3. Expression of matrix metalloproteinase-7 and tissue inhibitor of metalloproteinase-2 in human endometrial carcinoma

    Institute of Scientific and Technical Information of China (English)

    Chen Mei; Bo Nai-xiu; Huang Ya-jun; Dai Qi; Gong Li-mei

    2010-01-01

    Objective: To investigate the expression of matrix metalloproteinase-7 (MMP-7) and its tissue inhibitor (TIMP-2) in endometrial carcinoma and analyze their significance in endometrial cancer′s invasion and metastasis. Methods: Endometrial tissues were collected from 64 patients with endometrial carcinoma, 20 patients with endometrial hyperplasia and 20 normal women. The expressions of MMP-7, TIMP-2 in endometrium were measured by immuohistochemistry. Results: Expressions of MMP-7, TIMP-2 in endometrium of patients with endometrial carcinoma were significantly higher than those in normal endometrium (P<0.05). MMP-7 expression increased with surgical-pathological staging, depth of myometrial invasion, histologic grades and lymph node metastasis (P<0.05), while TIMP-2 expression was related to lymph node metastasis (P<0.05). TIMP-2 expression in endometrial cancer was significantly higher than that in hyperplastic endometrium (P<0.05). Expressions of TIMP-2 and MMP-7 in endometrium of patients with endometrial carcinoma were positively correlated (r=0.654, P<0.001). Conclusion: Highly expressed MMP-7 and TIMP-2 in endometrium may be related to development, invasion and metastasis of endometrial cancers.

  4. Muscle-derived Decellularised Extracellular Matrix Improves Functional Recovery in a Rat Latissimus Dorsi Muscle Defect Model

    Science.gov (United States)

    2013-12-01

    Piccolo R, et al. Experimental abdominal wall defect repaired with acellular matrix. Pediatr Surg Int 2002;18:327e31. 28. Willett NJ, Li MT, Uhrig BA...matrix (M-ECM). Methods: Ten percent of the mass of the latissimus dorsi (LD) was excised. Three groups were examined: 1) no repair of defect (DEF), 2...to develop therapies for ex- tremity10,11 or abdominal muscle repair.16 For the devel- opment of potential therapies for craniofacial muscle repair

  5. Matrix Metalloproteinase-9 and Haemozoin: Wedding Rings for Human Host and Plasmodium falciparum Parasite in Complicated Malaria

    Directory of Open Access Journals (Sweden)

    Mauro Prato

    2011-01-01

    Full Text Available It is generally accepted that the combination of both Plasmodium falciparum parasite and human host factors is involved in the pathogenesis of complicated severe malaria, including cerebral malaria (CM. Among parasite products, the malarial pigment haemozoin (HZ has been shown to impair the functions of mononuclear and endothelial cells. Different CM models were associated with enhanced levels of matrix metalloproteinases (MMPs, a family of proteolytic enzymes able to disrupt subendothelial basement membrane and tight junctions and shed, activate, or inactivate cytokines, chemokines, and other MMPs through cleavage from their precursors. Among MMPs, a good candidate for targeted therapy might be MMP-9, whose mRNA and protein expression enhancement as well as direct proenzyme activation by HZ have been recently investigated in a series of studies by our group and others. In the present paper the role of HZ and MMP-9 in complicated malaria, as well as their interactions, will be discussed.

  6. Extracellular Matrix Modulates Morphology, Growth, Oxidative Stress Response and Functionality of Human Skin Fibroblasts during Aging In Vitro

    DEFF Research Database (Denmark)

    Jørgensen, Peter; Rattan, Suresh

    2014-01-01

    The Hayflick system of cellular aging and replicative senescence in vitro has been used widely in both basic and applied research in biogerontology. The state of replicative senescence is generally considered to be irreversible, but is modifiable by genetic and environmental manipulations. Some...... recent observations indicate that replicative lifespan, senescence and functionality of cells in vitro can be significantly affected by the quality of the extra cellular matrix (ECM). Following up on those reports, here we show that using the ECM prepared from early passage young cells, partial...... rejuvenation of serially passaged human facial skin fibroblasts was possible in pre-senescent middle-aged cells, but not in fully senescent late passage cells. ECM from young cells improved the appearance, viability, stress tolerance and wound healing ability of skin fibroblasts. Furthermore, young ECM...

  7. Interactions of surface-displayed glycolytic enzymes of Mycoplasma pneumoniae with components of the human extracellular matrix.

    Science.gov (United States)

    Gründel, Anne; Jacobs, Enno; Dumke, Roger

    2016-12-01

    Mycoplasma pneumoniae is a major cause of community-acquired respiratory infections worldwide. Due to the strongly reduced genome, the number of virulence factors expressed by this cell wall-less pathogen is limited. To further understand the processes during host colonization, we investigated the interactions of the previously confirmed surface-located glycolytic enzymes of M. pneumoniae (pyruvate dehydrogenase A-C [PdhA-C], glyceraldehyde-3-phosphate dehydrogenase [GapA], lactate dehydrogenase [Ldh], phosphoglycerate mutase [Pgm], pyruvate kinase [Pyk] and transketolase [Tkt]) to the human extracellular matrix (ECM) proteins fibrinogen (Fn), fibronectin (Fc), lactoferrin (Lf), laminin (Ln) and vitronectin (Vc), respectively. Concentration-dependent interactions between Fn and Vc and all eight recombinant proteins derived from glycolytic enzymes, between Ln and PdhB-C, GapA, Ldh, Pgm, Pyk and Tkt, between Lf and PdhA-C, GapA and Pyk, and between Fc and PdhC and GapA were demonstrated. In most cases, these associations are significantly influenced by ionic forces and by polyclonal sera against recombinant proteins. In immunoblotting, the complex of human plasminogen, activator (tissue-type or urokinase plasminogen activator) and glycolytic enzyme was not able to degrade Fc, Lf and Ln, respectively. In contrast, degradation of Vc was confirmed in the presence of all eight enzymes tested. Our data suggest that the multifaceted associations of surface-localized glycolytic enzymes play a potential role in the adhesion and invasion processes during infection of human respiratory mucosa by M. pneumoniae.

  8. Effects of the enamel matrix derivative on the proliferation and odontogenic differentiation of human dental pulp cells.

    Science.gov (United States)

    Wang, Yuanyuan; Zhao, Yuming; Ge, Lihong

    2014-01-01

    The enamel matrix derivative (EMD) has a positive effect on the proliferation of human periodontal ligament cells and the healing of periodontal tissues. The aim of this study was to evaluate the effects of EMD on the proliferation and differentiation of human dental pulp cells (hDPCs) in vitro. hDPCs were isolated from human impacted third molars and cultured in vitro. After treatment with100μg/mL EMD, the proliferation of hDPCs was determined by a cell counting kit 8 (CCK8) assay. After incubation in EMD osteogenic induction medium for 14 days, the osteogenic differentiation of hDPCs was evaluated by alkaline phosphatase (ALP) activity, alizarin staining and the expression of osteogenesis-related genes. The EMD osteogenic induction medium enhanced the proliferation of hDPCs. After osteogenic induction, EMD increased the osteogenic potential of hDPCs, as measured by alkaline phosphatase activity and calcium accumulation; the expression levels of osteogenesis-related genes, such as ALP, DSPP, BMP, and OPN were also upregulated. In addition, the expression levels of odontogenesis-related transcription factors Osterix and Runx2 were upregulated. EMD could enhance the mineralization of hDPSCs upregulated the expression of markers for odontoblast/osteoblast-like cells. Furth