Preparation and Evaluation of Gelatin-Chitosan-Nanobioglass 3D Porous Scaffold for Bone Tissue Engineering

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Maji, Kanchan; Dasgupta, Sudip; Pramanik, Krishna; Bissoyi, Akalabya

2016-01-01

The aim of the present study was to prepare and characterize bioglass-natural biopolymer based composite scaffold and evaluate its bone regeneration ability. Bioactive glass nanoparticles (58S) in the size range of 20?30?nm were synthesized using sol-gel method. Porous scaffolds with varying bioglass composition from 10 to 30?wt% in chitosan, gelatin matrix were fabricated using the method of freeze drying of its slurry at 40?wt% solids loading. Samples were cross-linked with glutaraldehyde t...

Fabrication of mineralized electrospun PLGA and PLGA/gelatin nanofibers and their potential in bone tissue engineering

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Meng, Z.X. [Center for Biomedical Materials and Engineering, Harbin Engineering University, Harbin 150001 (China); Li, H.F. [Department of Materials Science and Engineering, College of Engineering, Peking University, Beijing 100871 (China); Sun, Z.Z. [Center for Biomedical Materials and Engineering, Harbin Engineering University, Harbin 150001 (China); Zheng, W., E-mail: zhengwei@hrbeu.edu.cn [Center for Biomedical Materials and Engineering, Harbin Engineering University, Harbin 150001 (China); Zheng, Y.F., E-mail: yfzheng@pku.edu.cn [Center for Biomedical Materials and Engineering, Harbin Engineering University, Harbin 150001 (China); Department of Materials Science and Engineering, College of Engineering, Peking University, Beijing 100871 (China)

2013-03-01

Surface mineralization is an effective method to produce calcium phosphate apatite coating on the surface of bone tissue scaffold which could create an osteophilic environment similar to the natural extracellular matrix for bone cells. In this study, we prepared mineralized poly(D,L-lactide-co-glycolide) (PLGA) and PLGA/gelatin electrospun nanofibers via depositing calcium phosphate apatite coating on the surface of these nanofibers to fabricate bone tissue engineering scaffolds by concentrated simulated body fluid method, supersaturated calcification solution method and alternate soaking method. The apatite products were characterized by the scanning electron microscopy (SEM), Fourier transform-infrared spectroscopy (FT-IR), and X-ray diffractometry (XRD) methods. A large amount of calcium phosphate apatite composed of dicalcium phosphate dihydrate (DCPD), hydroxyapatite (HA) and octacalcium phosphate (OCP) was deposited on the surface of resulting nanofibers in short times via three mineralizing methods. A larger amount of calcium phosphate was deposited on the surface of PLGA/gelatin nanofibers rather than PLGA nanofibers because gelatin acted as nucleation center for the formation of calcium phosphate. The cell culture experiments revealed that the difference of morphology and components of calcium phosphate apatite did not show much influence on the cell adhesion, proliferation and activity. - Highlights: Black-Right-Pointing-Pointer Ca-P phases were coated on PLGA/gelatin electrospun nanofiber membranes within 3 h. Black-Right-Pointing-Pointer Ca-P coatings prepared by 3 methods exhibited different structures and components. Black-Right-Pointing-Pointer The Ca-P coating weight increase depends on the apatite nucleation velocity. Black-Right-Pointing-Pointer Surface hydrophilicity enhanced the velocity and quantity of apatite nucleation. Black-Right-Pointing-Pointer The resulting Ca-P apatite coatings exhibit good biocompatibility to MG63 cells.

Assessment of Matrix Metalloproteinases by Gelatin Zymography.

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Cathcart, Jillian

2016-01-01

Matrix metalloproteinases are endopeptidases responsible for remodeling of the extracellular matrix and have been identified as critical contributors to breast cancer progression. Gelatin zymography is a valuable tool which allows the analysis of MMP expression. In this approach, enzymes are resolved electrophoretically on a sodium dodecyl sulfate-polyacrylamide gel copolymerized with the substrate for the MMP of interest. Post electrophoresis, the enzymes are refolded in order for proteolysis of the incorporated substrate to occur. This assay yields valuable information about MMP isoforms or changes in activation and can be used to analyze the role of MMPs in normal versus pathological conditions.

Gelatin/nano-hydroxyapatite hydrogel scaffold prepared by sol-gel technology as filler to repair bone defects.

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Raucci, Maria Grazia; Demitri, Christian; Soriente, Alessandra; Fasolino, Ines; Sannino, Alessandro; Ambrosio, Luigi

2018-07-01

This study reports on the development of a scaffold with a gradient of bioactive solid signal embedded in the biodegradable polymer matrix by combining a sol-gel approach and freeze-drying technology. The chemical approach based on the sol-gel transition of calcium phosphates ensures the particles dispersion into the gelatin matrix and a direct control of interaction among COOH gelatin /Ca 2+ ions. Morphological analysis demonstrated that on the basis of the amount of inorganic component and by using specific process conditions, it is possible to control the spatial distribution of nanoparticles around the gelatin helix. In fact, methodology and formulations were able to discriminate between the different hydroxyapatite concentrations and their respective morphology. The good biological response represented by good cell attachment, proliferation and increased levels of alkaline phosphatase as an indicator of osteoblastic differentiation of human mesenchymal stem cells toward the osteogenic lineage, demonstrating the effect of bioactive solid signals on cellular behavior. Furthermore, the inhibition of reactive oxygen species production by composite materials predicted potential anti-inflammatory properties of scaffolds thus confirming their biocompatibility. Indeed, these interesting biological results suggest good potential application of this scaffold as filler to repair bone defects. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 2007-2019, 2018. © 2018 Wiley Periodicals, Inc.

In vitro and in vivo investigations on bone regeneration potential of laminated hydroxyapatite/gelatin nanocomposite scaffold along with DBM

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Tavakol, Shima [School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Department of Medical Nanotechnology (Iran, Islamic Republic of); Ragerdi Kashani, Iraj [School of Medicine, Tehran University of Medical Sciences, Department of Anatomy (Iran, Islamic Republic of); Azami, Mahmood [School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Department of Tissue Engineering (Iran, Islamic Republic of); Khoshzaban, Ahad [Tehran University of Medical Sciences, Iranian Tissue Bank Research and Preparation Center (Iran, Islamic Republic of); Tavakol, Behnaz [Kashan University of Medical Sciences, Department of Medicine (Iran, Islamic Republic of); Kharrazi, Sharmin [School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Department of Medical Nanotechnology (Iran, Islamic Republic of); Ebrahimi, Somayeh [University of Tarbiat Moallem, Department of Biology, Faculty of Sciences (Iran, Islamic Republic of); Rezayat Sorkhabadi, Seyed Mahdi, E-mail: sh_tavakol@razi.tums.ac.ir [School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Department of Medical Nanotechnology (Iran, Islamic Republic of)

2012-12-15

Bone regeneration ability of a scaffold strongly depends on its structure and the size of its components. In this study, a nanostructured scaffold was designed for bone repair using nano hydroxyapatite (nHA) (8-16 nm Multiplication-Sign 50-80 nm) and gelatin (GEL) as main components. In vitro investigations of calcium matrix deposition and gene expression of the seeded cells for this scaffold, demineralized bone matrix (DBM), scaffold plus DBM, and the control group were carried out. Bone regeneration in rat calvarium with critical defect size after 1, 4, and 8 weeks post implantation was investigated. The calcium matrix depositions by the osteoblast and RUNX2, ALP, osteonectin, and osteocalcin gene expression in scaffold were more significant than in other groups. Histomorphometry analysis confirmed in vitro results. In vitro and in vivo bone regeneration were least in scaffold plus DBM group. Enhanced effects in scaffold could be attributed to the shape and size of nHA particles and good architecture of the scaffold. Reduction of bone regeneration might be due to tight bonding of BMPs and nHA particles in the third group. Results obtained from this study confirmed that nano-scale size of the main components and the scaffold architecture (pore diameter, interconnectivity pores, etc.) have significant effects on bone regeneration ability of the scaffold and are important parameters in designing a temporary bone substitute.

In vitro and in vivo investigations on bone regeneration potential of laminated hydroxyapatite/gelatin nanocomposite scaffold along with DBM

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Tavakol, Shima; Ragerdi Kashani, Iraj; Azami, Mahmood; Khoshzaban, Ahad; Tavakol, Behnaz; Kharrazi, Sharmin; Ebrahimi, Somayeh; Rezayat Sorkhabadi, Seyed Mahdi

2012-01-01

Bone regeneration ability of a scaffold strongly depends on its structure and the size of its components. In this study, a nanostructured scaffold was designed for bone repair using nano hydroxyapatite (nHA) (8–16 nm × 50–80 nm) and gelatin (GEL) as main components. In vitro investigations of calcium matrix deposition and gene expression of the seeded cells for this scaffold, demineralized bone matrix (DBM), scaffold plus DBM, and the control group were carried out. Bone regeneration in rat calvarium with critical defect size after 1, 4, and 8 weeks post implantation was investigated. The calcium matrix depositions by the osteoblast and RUNX2, ALP, osteonectin, and osteocalcin gene expression in scaffold were more significant than in other groups. Histomorphometry analysis confirmed in vitro results. In vitro and in vivo bone regeneration were least in scaffold plus DBM group. Enhanced effects in scaffold could be attributed to the shape and size of nHA particles and good architecture of the scaffold. Reduction of bone regeneration might be due to tight bonding of BMPs and nHA particles in the third group. Results obtained from this study confirmed that nano-scale size of the main components and the scaffold architecture (pore diameter, interconnectivity pores, etc.) have significant effects on bone regeneration ability of the scaffold and are important parameters in designing a temporary bone substitute.

Gelatin Tight-Coated Poly(lactide-co-glycolide Scaffold Incorporating rhBMP-2 for Bone Tissue Engineering

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Juan Wang

2015-03-01

Full Text Available Surface coating is the simplest surface modification. However, bioactive molecules can not spread well on the commonly used polylactone-type skeletons; thus, the surface coatings of biomolecules are typically unstable due to the weak interaction between the polymer and the bioactive molecules. In this study, a special type of poly(lactide-co-glycolide (PLGA-based scaffold with a loosened skeleton was fabricated by phase separation, which allowed gelatin molecules to more readily diffuse throughout the structure. In this application, gelatin modified both the internal substrate and external surface. After cross-linking with glutaraldehyde, the surface layer gelatin was tightly bound to the diffused gelatin, thereby preventing the surface layer gelatin coating from falling off within 14 days. After gelatin modification, PLGA scaffold demonstrated enhanced hydrophilicity and improved mechanical properties (i.e., increased compression strength and elastic modulus in dry and wet states. Furthermore, a sustained release profile of recombinant human bone morphogenetic protein-2 (rhBMP-2 was achieved in the coated scaffold. The coated scaffold also supported the in vitro attachment, proliferation, and osteogenesis of rabbit bone mesenchymal stem cells (BMSCs, indicating the bioactivity of rhBMP-2. These results collectively demonstrate that the cross-linked-gelatin-coated porous PLGA scaffold incorporating bioactive molecules is a promising candidate for bone tissue regeneration.

Coating electrospun poly(epsilon-caprolactone) fibers with gelatin and calcium phosphate and their use as biomimetic scaffolds for bone tissue engineering.

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Li, Xiaoran; Xie, Jingwei; Yuan, Xiaoyan; Xia, Younan

2008-12-16

Electrospinning was employed to fabricate fibrous scaffolds of poly(epsilon-caprolactone) in the form of nonwoven mats. The surfaces of the fibers were then coated with gelatin through layer-by-layer self-assembly, followed by functionalization with a uniform coating of bonelike calcium phosphate by mineralization in the 10 times concentrated simulated body fluid for 2 h. Transmission electron microscopy, water contact angle, and scanning electron microscopy measurements confirmed the presence of gelatin and calcium phosphate coating layers, and X-ray diffraction results suggested that the deposited mineral phase was a mixture of dicalcium phosphate dehydrate (a precursor to apatite) and apatite. It was also demonstrated that the incorporation of gelatin promoted nucleation and growth of calcium phosphate. The porous scaffolds could mimic the structure, composition, and biological function of bone extracellular matrix. It was found that the preosteoblastic MC3T3-E1 cells attached, spread, and proliferated well with a flat morphology on the mineralized scaffolds. The proliferation rate of the cells on the mineralized scaffolds was significantly higher (by 1.9-fold) than that on the pristine fibrous scaffolds after culture for 7 days. These results indicated that the hybrid system containing poly(epsilon-caprolactone), gelatin, and calcium phosphate could serve as a new class of biomimetic scaffolds for bone tissue engineering.

Study and characterization of powder mackerel (Scomberomorus commerson) bone gelatin through hydrolysis of hydrochloric acid

Science.gov (United States)

Mardawati, E.; Sugandi, H.; Kayaputri, I. L.; Cahyana, Y.; Wira, D. W.; Pujianto, T.; Kastaman, R.

2018-02-01

Gelatin is one of the most common food additives in the food and beverage industry. Gelatin is generally made of leather or pig bones, causing concerns about the halal and safety of its product. Mackerel fish bone (Scomberomorus commerson) is a waste fish that has not been utilized well and it contains 18.6% of collagen so that it can be made into gelatin. The purpose of this research is to know the relation between HCl concentration with physical and chemical characteristics of gelatin and to know the best HCl concentration for gelatin production. Based on the physical and chemical analysis of gelatin, it is known that the concentration of hydrochloric acid influences the yield, viscosity, gel strength and pH produced. The higher HCl concentration there will be decrease in the pH value, gel strength, viscosity and protein. The yield will rise to the optimum point then decrease with respect to the high HCl concentration. Gelatin with 2% HCl concentration was the best treatment, with pH value 3.83, viscosity 3.65cP, gel strength 190.50 blooms which fulfilled British Standard, yield 10.16%, protein content 43.34%. It has functional group such as amino acids glycine, proline and hydroxyproline and 15 other amino acids, the gelatin group uptake in the region of amide wave numbers A, amides I, II and III, with a gelatin molecular weight of 290.35 g/mol.

Preparation and bioactivity evaluation of hydroxyapatite-titania/chitosan-gelatin polymeric biocomposites

International Nuclear Information System (INIS)

Mohamed, Khaled R.; Mostafa, Amani A.

2008-01-01

Biocomposites consisting of hydroxyapatite (HA) and natural polymers such as collagen, chitosan, chitin,and gelatin have been extensively investigated. However, studies on the combination of HA and titania with chitosan and gelatin have not been conducted yet. Novel biodegradable hydroxyapatite-titania/chitosan-gelatin polymeric composites were fabricated. In this work, our results are concerning with the preparation and characterization of HA powder and HA filler containing titania powder (10 and 30%) with a chitosan and gelatin copolymer matrix. The present research focuses on characterizing the structure of this novel class of biocomposites. Thermogravimetric analysis (TGA), X-ray diffraction (XRD), and Fourier Transformed Infrared Spectroscopy (FT-IR), Scanning electron microscopy (SEM-EDAX) were employed to assess the produced composites. The mechanical properties in terms of compressive strength and hardness test were also investigated. The in vitro study in simulated body fluid (SBF) was performed to assess the bioactivity of composites. The results proved that apatite resembling natural bone are formed faster and greater in the case the composite of HA containing 10% titania into chitosan-gelatin polymeric matrix when they are soaked in a simulated body fluid (SBF) than the composite containing 30% titania. The biocomposites containing HA with 10% titania are expected to be attractive for bioapplications as bone substitutes and scaffolds for tissue engineering in future

[Fabrication of porous poly lactic acid-bone matrix gelatin composite bioactive material and its osteoinductive activity].

Science.gov (United States)

Zhang, Yumin; Li, Baoxing; Li, Ji

2007-02-01

To fabricate a novel porous bioactive composite biomaterial consisting of poly lactic acid (PLA)-bone matrix gelatin (BMG) by using the supercritical carbon dioxide fluid technique (SC-CO2) and to evaluate its osteoinductive activity. The cortical bones selected from healthy adult donors were processed into BMG by the defatting, demineralizing, and deproteinizing processes. PLA and BMG were mixed at a volume radio of 3 : 1; then, the PLA-BMG mixed material and the pure PLA material were respectively placed in the supercritical carbon dioxide reaction kettles, and were respectively added by the NaCl particles 100-200 microm in diameter for the porosity of the materials so that the porous PLA-BMG composite material and the porous PLA composite material could be formed. The mouse osteoblast-like MC3T3-E1 cells were cultured in the dulbecco's modified eagle medium (DMEM) supplemented with 10% fetal bovine serum. Then, 20 microl of the MC3T3-E1 cell suspensions containing 2 X 10(6) cells /ml were delivered into the culturing plate (24 wells/plate) made of the different materials, which were co-cultured for 2 weeks. In the PLA-BMG group, 100 microg of the crushed PLA-BMG material was contained in each well; in the PLA group, 100 microg of the crushed PLA material was contained in each well; and in the DMEM group, only DMEM was contained, which served as the control group. There were 6 wells in each group. The quantitative analysis on the calcification area was performed by the staining of the alizarin red S. The co-cultured cells were harvested and lysated in 1 ml of 0. 2% Nonidet P-40 by the ultrasonic lysating technique. Then, the ALP activity and the Ca content were measured according to the illuminations of the reagent kits. The porous PLA-BMG composite material showed a good homological porosity with a pore diameter of 50-150 microm and a good connectivity between the pores. The ALP activity, the Ca content, and the calcification area were significantly greater in

Mineralization and Characterization of Composite Lyophilized Gelatin Sponges Intended for Early Bone Regeneration

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Isaac Rodriguez

2014-01-01

Full Text Available The application of freeze-dried gelatin sponges as alternative bone grafting substitutes has many advantages, including the ability to swell, high porosity, tailorable degradation, and versatility to incorporate multiple components such as growth factors and nanofillers. The purpose of this study was to mineralize (M and further characterize 1-Ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC cross-linked gelatin sponges enhanced with preparations rich in growth factors, hydroxyapatite, and chitin whiskers (PHCE. Sponges were characterized for their swelling and in vitro mineralization potential, surface characteristics, protein release, mechanical properties, and MG-63 cell attachment and infiltration. All sponges swelled up to 50% of their original volume upon hydration. Scanning electron microscopy showed sparse mineral deposition for gelatin-M scaffolds while PHCE-M scaffolds exhibited more uniform mineral nucleation. Over 21 days, PHCE-M scaffolds cumulatively released significantly more (30% of its initial protein content than all other scaffolds. PHCE-M scaffolds reported lower modulus values (1.3–1.6 MPa when compared to gelatin control scaffolds (1.6–3.2 MPa. Increased cell attachment and infiltration was noticed on PHCE and PHCE-M scaffolds. The results of the study demonstrate the enhanced performance of PHCE and PHCE-M scaffolds to serve as bone healing scaffolds. Their potential to release incorporated factors, comparable composition/mechanical properties to tissues developed in the early stages of bone healing, and enhanced initial cellular response make them suitable for further studies evaluating more complex cellular interactions.

Sustained delivery of biomolecules from gelatin carriers for applications in bone regeneration.

Science.gov (United States)

Song, Jiankang; Leeuwenburgh, Sander Cg

2014-08-01

Local delivery of therapeutic biomolecules to stimulate bone regeneration has matured considerably during the past decades, but control over the release of these biomolecules still remains a major challenge. To this end, suitable carriers that allow for tunable spatial and temporal delivery of biomolecules need to be developed. Gelatin is one of the most widely used natural polymers for the controlled and sustained delivery of biomolecules because of its biodegradability, biocompatibility, biosafety and cost-effectiveness. The current study reviews the applications of gelatin as carriers in form of bulk hydrogels, microspheres, nanospheres, colloidal gels and composites for the programmed delivery of commonly used biomolecules for applications in bone regeneration with a specific focus on the relationship between carrier properties and delivery characteristics.

Electrospun Gelatin/β-TCP Composite Nanofibers Enhance Osteogenic Differentiation of BMSCs and In Vivo Bone Formation by Activating Ca2+-Sensing Receptor Signaling

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Xuehui Zhang

2015-01-01

Full Text Available Calcium phosphate- (CaP- based composite scaffolds have been used extensively for the bone regeneration in bone tissue engineering. Previously, we developed a biomimetic composite nanofibrous membrane of gelatin/β-tricalcium phosphate (TCP and confirmed their biological activity in vitro and bone regeneration in vivo. However, how these composite nanofibers promote the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs is unknown. Here, gelatin/β-TCP composite nanofibers were fabricated by incorporating 20 wt% β-TCP nanoparticles into electrospun gelatin nanofibers. Electron microscopy showed that the composite β-TCP nanofibers had a nonwoven structure with a porous network and a rough surface. Spectral analyses confirmed the presence and chemical stability of the β-TCP and gelatin components. Compared with pure gelatin nanofibers, gelatin/β-TCP composite nanofibers caused increased cell attachment, proliferation, alkaline phosphatase activity, and osteogenic gene expression in rat BMSCs. Interestingly, the expression level of the calcium-sensing receptor (CaSR was significantly higher on the composite nanofibrous scaffolds than on pure gelatin. For rat calvarial critical sized defects, more extensive osteogenesis and neovascularization occurred in the composite scaffolds group compared with the gelatin group. Thus, gelatin/β-TCP composite scaffolds promote osteogenic differentiation of BMSCs in vitro and bone regeneration in vivo by activating Ca2+-sensing receptor signaling.

Zymographic analysis using gelatin-coated film of the effect of etanercept on the extracellular matrix-degrading activity in synovial fluids of rheumatoid arthritis patients.

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Kamataki, Akihisa; Ishida, Mutsuko; Komagamine, Masataka; Yoshida, Masaaki; Ando, Takanobu; Sawai, Takashi

2016-04-01

Rheumatoid arthritis (RA) is a chronic inflammatory disease. Most RA patients develop cartilage and bone destruction, and various proteinases are involved in the destruction of extracellular matrix of cartilage and bone. The aim of this study is to evaluate the utility of our newly developed method to measure total gelatinolytic activity. We adopted this method for measurement in synovial fluid from RA patients treated by the anti-rheumatic drug etanercept (ETN), a recombinant human soluble tumor necrosis factor receptor fusion protein, and compared the findings with clinical and laboratory data. Enzymatic activity of synovial fluid was analyzed by zymography using gelatin-coated film, and compared with the index of Disease Activity Score of 28 joints - C-reactive protein (DAS28-CRP), CRP and matrix metalloproteinase (MMP)-3 level before and after ETN therapy. Synovial fluids of 19 patients were collected before and after administration of ETN therapy. In nine of 19 patients, who showed a decrease in gelatin-degrading activity in synovial fluid, the index of DAS28-CRP (4.85-2.85, ΔDAS = -2.00) and CRP (3.30-0.94 mg/dL, ΔCRP = -2.36) was alleviated after ETN therapy, while cases with no change or an increase in gelatin-degrading activity showed a modest improvement in clinical data: DAS28-CRP (4.23-3.38, ΔDAS = -0.85) and CRP (1.70-0.74 mg/dL, ΔCRP = -0.96). Our newly developed method for measurement of gelatin-degrading activity in synovial fluid from RA patients is highly practicable and useful for predicting the effect of ETN therapy. © 2013 Asia Pacific League of Associations for Rheumatology and Wiley Publishing Asia Pty Ltd.

Characteristic study of chitosan addition in Tilapia (Oreochromis niloticus) bone based gelatin film

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Atmaka, W.; Yudhistira, B.; Putro, M. I. S.

2018-03-01

Tilapia (Oreochromis niloticus) is one of popular fish species in Indonesia. The high number in tilapia’s production and export of tilapia resulting in the increase of bone fish waste. an attempt to decrease the amount of the aforementioned waste, the fish bones were turned into gelatine. The gelatine produced from this waste can be put to good use by turning it into edible film due to its high water resistance and low tensile strength value. However, in order to make a proper film, both the water resistance and the tensile strength value needs another appropriate additional biopolymer. In this case, the appropriate biopolymer needed both to form the film and to repair its characteristics is chitosan. The purpose of this research is to find out the effect of the chitosan addition on the tilapia bone based gelatine film. The research used several mixtures of gelatine (G) and chitosan (C) with the following ratio: G100:C0 (GC1), G75:C25 (GC2), G50:C50 (GC3), G25:C75 (GC4), G0:C100 (GC5). ANOVA results (Psolubility, tensile strength, elongation at break, Fourier Transform Infrared (FTIR), color a, and color b but no significant effects on the vapor permeability and color L. The best result is shown on GC2 with thickness 0.119 mm; solubility 74.95%; tensile strength 2.635 Mpa; elongation at break 68.26%; water vapor permeability 5.897 g/h m2 and FTIR. The parameters in GC2 shows good compatibility between the two biopolymers.

Preparation of a biomimetic composite scaffold from gelatin/collagen and bioactive glass fibers for bone tissue engineering

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Sharifi, Esmaeel; Azami, Mahmoud [Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Kajbafzadeh, Abdol-Mohammad [Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Pediatric Urology Research Center, Section of Tissue Engineering and Stem Cells Therapy, Department of Pediatric Urology, Children' s Hospital Medical Center, Tehran, Iran (IRI) (Iran, Islamic Republic of); Moztarzadeh, Fatollah [Department of Biomedical Engineering, Amirkabir University of Technology (Tehran Polytechnic), Tehran (Iran, Islamic Republic of); Faridi-Majidi, Reza [Department of Medical Nanotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Shamousi, Atefeh; Karimi, Roya [Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Ai, Jafar, E-mail: jafar_ai@tums.ac.ir [Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Brain and Spinal Injury Research Center (BASIR), Imam Khomeini Hospital, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of)

2016-02-01

Bone tissue is a composite material made of organic and inorganic components. Bone tissue engineering requires scaffolds that mimic bone nature in chemical and mechanical properties. This study proposes a novel method for preparing composite scaffolds that uses sub-micron bioglass fibers as the organic phase and gelatin/collagen as the inorganic phase. The scaffolds were constructed by using freeze drying and electro spinning methods and their mechanical properties were enhanced by using genipin crosslinking agent. Electron microscopy micrographs showed that the structure of composite scaffolds were porous with pore diameters of approximately 70–200 μm, this was again confirmed by mercury porosimetery. These pores are suitable for osteoblast growth. The diameters of the fibers were approximately 150–450 nm. Structural analysis confirmed the formation of desirable phases of sub-micron bioglass fibers. Cellular biocompatibility tests illustrated that scaffolds containing copper ion in the bioglass structure had more cell growth and osteoblast attachment in comparison to copper-free scaffolds. - Highlights: • Fabrication of 45S5 sub-micron bioglass fiber using electrospinning method. • Production of copper doped submicron bioglass fibers on 45S5 bioglass base by electrospinning sol gel route method. • Incorporation of bioglass/Cu-bioglass sub-micron fibers into gelatin/collagen matrix to form biomimetic composite scaffold which were non-cytotoxic according to MTT assay. • Discovering that copper can decrease the glass transition temperatures and enhance osteoblast cell adhesion and viability.

Preparation of a biomimetic composite scaffold from gelatin/collagen and bioactive glass fibers for bone tissue engineering

International Nuclear Information System (INIS)

Sharifi, Esmaeel; Azami, Mahmoud; Kajbafzadeh, Abdol-Mohammad; Moztarzadeh, Fatollah; Faridi-Majidi, Reza; Shamousi, Atefeh; Karimi, Roya; Ai, Jafar

2016-01-01

Bone tissue is a composite material made of organic and inorganic components. Bone tissue engineering requires scaffolds that mimic bone nature in chemical and mechanical properties. This study proposes a novel method for preparing composite scaffolds that uses sub-micron bioglass fibers as the organic phase and gelatin/collagen as the inorganic phase. The scaffolds were constructed by using freeze drying and electro spinning methods and their mechanical properties were enhanced by using genipin crosslinking agent. Electron microscopy micrographs showed that the structure of composite scaffolds were porous with pore diameters of approximately 70–200 μm, this was again confirmed by mercury porosimetery. These pores are suitable for osteoblast growth. The diameters of the fibers were approximately 150–450 nm. Structural analysis confirmed the formation of desirable phases of sub-micron bioglass fibers. Cellular biocompatibility tests illustrated that scaffolds containing copper ion in the bioglass structure had more cell growth and osteoblast attachment in comparison to copper-free scaffolds. - Highlights: • Fabrication of 45S5 sub-micron bioglass fiber using electrospinning method. • Production of copper doped submicron bioglass fibers on 45S5 bioglass base by electrospinning sol gel route method. • Incorporation of bioglass/Cu-bioglass sub-micron fibers into gelatin/collagen matrix to form biomimetic composite scaffold which were non-cytotoxic according to MTT assay. • Discovering that copper can decrease the glass transition temperatures and enhance osteoblast cell adhesion and viability.

Enhanced Critical Size Defect Repair in Rabbit Mandible by Electrospun Gelatin/β-TCP Composite Nanofibrous Membranes

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Mingming Xu

2015-01-01

Full Text Available The design and fabrication of biodegradable barrier membranes with satisfactory structure and composition remain a considerable challenge for periodontal tissue regeneration. We have developed a biomimetic nanofibrous membrane made from a composite of gelatin and β-tricalcium phosphate (β-TCP. We previously confirmed the in vitro biological performance of the membrane material, but the efficacy of the membranes in promoting bone repair in situ has not yet been examined. Gelatin/β-TCP composite nanofibers were fabricated by incorporation of 20 wt.% β-TCP nanoparticles into electrospun gelatin nanofibers. Electron microscopy showed that the composite membranes presented a nonwoven structure with an interconnected porous network and had a rough surface due to the β-TCP nanoparticles, which were distributed widely and uniformly throughout the gelatin-fiber matrix. The repair efficacy of rabbit mandible defects implanted with bone substitute (Bio-Oss and covered with the gelatin/β-TCP composite nanofibrous membrane was evaluated in comparison with pure gelatin nanofibrous membrane. Gross observation, histological examination, and immunohistochemical analysis showed that new bone formation and defect closure were significantly enhanced by the composite membranes compared to the pure gelatin ones. From these results, we conclude that nanofibrous gelatin/β-TCP composite membranes could serve as effective barrier membranes for guided tissue regeneration.

Production and characterization of chitosan/gelatin/β-TCP scaffolds for improved bone tissue regeneration

Energy Technology Data Exchange (ETDEWEB)

Serra, I.R.; Fradique, R.; Vallejo, M.C.S.; Correia, T.R.; Miguel, S.P.; Correia, I.J., E-mail: icorreia@ubi.pt

2015-10-01

Recently, bone tissue engineering emerged as a viable therapeutic alternative, comprising bone implants and new personalized scaffolds to be used in bone replacement and regeneration. In this study, biocompatible scaffolds were produced by freeze-drying, using different formulations (chitosan, chitosan/gelatin, chitosan/β-TCP and chitosan/gelatin/β-TCP) to be used as temporary templates during bone tissue regeneration. Sample characterization was performed through attenuated total reflectance-Fourier transform infrared spectroscopy, X-ray diffraction and energy dispersive spectroscopy analysis. Mechanical characterization and porosity analysis were performed through uniaxial compression test and liquid displacement method, respectively. In vitro studies were also done to evaluate the biomineralization activity and the cytotoxic profile of the scaffolds. Scanning electron and confocal microscopy analysis were used to study cell adhesion and proliferation at the scaffold surface and within their structure. Moreover, the antibacterial activity of the scaffolds was also evaluated through the agar diffusion method. Overall, the results obtained revealed that the produced scaffolds are bioactive and biocompatible, allow cell internalization and show antimicrobial activity against Staphylococcus aureus. Such, make these 3D structures as potential candidates for being used on the bone tissue regeneration, since they promote cell adhesion and proliferation and also prevent biofilm development at their surfaces, which is usually the main cause of implant failure. - Highlights: • Production of 3D scaffolds composed by chitosan/gelatin/β-TCP by freeze-drying for bone regeneration • Physicochemical characterization of the bone substitutes by SEM, FTIR, XRD and EDS • Evaluation of the cytotoxic profile and antibacterial activity of the 3D structures through in vitro assays.

Production and characterization of chitosan/gelatin/β-TCP scaffolds for improved bone tissue regeneration

International Nuclear Information System (INIS)

Serra, I.R.; Fradique, R.; Vallejo, M.C.S.; Correia, T.R.; Miguel, S.P.; Correia, I.J.

2015-01-01

Recently, bone tissue engineering emerged as a viable therapeutic alternative, comprising bone implants and new personalized scaffolds to be used in bone replacement and regeneration. In this study, biocompatible scaffolds were produced by freeze-drying, using different formulations (chitosan, chitosan/gelatin, chitosan/β-TCP and chitosan/gelatin/β-TCP) to be used as temporary templates during bone tissue regeneration. Sample characterization was performed through attenuated total reflectance-Fourier transform infrared spectroscopy, X-ray diffraction and energy dispersive spectroscopy analysis. Mechanical characterization and porosity analysis were performed through uniaxial compression test and liquid displacement method, respectively. In vitro studies were also done to evaluate the biomineralization activity and the cytotoxic profile of the scaffolds. Scanning electron and confocal microscopy analysis were used to study cell adhesion and proliferation at the scaffold surface and within their structure. Moreover, the antibacterial activity of the scaffolds was also evaluated through the agar diffusion method. Overall, the results obtained revealed that the produced scaffolds are bioactive and biocompatible, allow cell internalization and show antimicrobial activity against Staphylococcus aureus. Such, make these 3D structures as potential candidates for being used on the bone tissue regeneration, since they promote cell adhesion and proliferation and also prevent biofilm development at their surfaces, which is usually the main cause of implant failure. - Highlights: • Production of 3D scaffolds composed by chitosan/gelatin/β-TCP by freeze-drying for bone regeneration • Physicochemical characterization of the bone substitutes by SEM, FTIR, XRD and EDS • Evaluation of the cytotoxic profile and antibacterial activity of the 3D structures through in vitro assays

Isolation of fish skin and bone gelatin from tilapia (Oreochromis niloticus): Response surface approach

Science.gov (United States)

Arpi, N.; Fahrizal; Novita, M.

2018-03-01

In this study, gelatin from fish collagen, as one of halal sources, was extracted from tilapia (Oreochromis niloticus) skin and bone, by using Response Surface Methodology to optimize gelatin extraction conditions. Concentrations of alkaline NaOH and acid HCl, in the pretreatment process, and temperatures in extraction process were chosen as independent variables, while dependent variables were yield, gel strength, and emulsion activity index (EAI). The result of investigation showed that lower NaOH pretreatment concentrations provided proper pH extraction conditions which combine with higher extraction temperatures resulted in high gelatin yield. However, gelatin emulsion activity index increased proportionally to the decreased in NaOH concentrations and extraction temperatures. No significant effect of the three independent variables on the gelatin gel strength. RSM optimization process resulted in optimum gelatin extraction process conditions using alkaline NaOH concentration of 0.77 N, acid HCl of 0.59 N, and extraction temperature of 66.80 °C. The optimal solution formula had optimization targets of 94.38%.

Viscoelastic and Functional Properties of Cod-Bone Gelatin in the Presence of Xylitol and Stevioside

Directory of Open Access Journals (Sweden)

Linyu Nian

2018-05-01

Full Text Available The physical, rheological, structural and functional properties of cod bone gelatin (CBG with various concentrations (0, 2, 4, 6, 10, and 15% of low-calorie sweeteners [xylitol (X and stevioside (S] to form gels were investigated. The gel strength of CBGX increased with increased xylitol due presumably to hydrogen bonds between xylitol and gelatin, but with CBGS the highest gel strength occurred when S concentration was 4%. Viscosity of CBGS samples were higher than CBGX due to S's high molecular mass. The viscoelasticity (G′ and G′′, foaming capacity and fat binding capacity of CBGX were higher while foam stability was lower. The emulsion activity and emulsion stability of CBGX were a little lower than CBGS at the same concentration. The structure of X is linear making it easier to form a dense three-dimensional network structure, while the complex cyclic structure of S had more difficulty forming a network structure with cod bone gelatin. Therefore, X may be a better choice for sweetening gelatin gels.

Multifunctional nano-hydroxyapatite and alginate/gelatin based sticky gel composites for potential bone regeneration

International Nuclear Information System (INIS)

Cai, Yurong; Yu, Juhong; Kundu, Subhas C.; Yao, Juming

2016-01-01

To improve the fixations of the implant and implant-bone integration after joint arthroplasty from locally preventing inflammation and promoting the bone regeneration, we design a multifunctional biomaterial consisting of recombinant human bone morphogenetic protein 2 (rhBMP-2) and antibiotic loaded nano-hydroxyapatite with an alginate/gelatin sticky gel. We investigate its role for the prevention of the inflammation and possibility of inducing a new bone growth along with its adhesive ability. The stickiness exists in the composite, which may help to fix itself on the bone fracture surface. The composite sustains the antibacterial effect and promotes the proliferation and differentiation of MG63 cells in vitro. In vivo experimentation also shows that the composite gel has a role for the reduction of inflammation. It enhances the formation of new bone and blood vessels compared to both the sole rhBMP-2 and non-rhBMP-2/antibiotic loaded composite gels. The multifunctional composite provides a promising material for the prosthetic and bone tissue regeneration. - Highlights: • Multifunctional nanohydroxyapatite composite is fabricated. • The composite consists of nHAP, growth factor, antibiotic and alginate/gelatin gel. • The composite shows antibacterial effect and good cytocompatibility. • No adverse effect to the cells tested in vitro and in vivo.

Multifunctional nano-hydroxyapatite and alginate/gelatin based sticky gel composites for potential bone regeneration

Energy Technology Data Exchange (ETDEWEB)

Cai, Yurong; Yu, Juhong [The Key Laboratory of Advanced Textile Materials and Manufacturing Technology of Ministry of Education, National Engineering Lab of Textile Fiber Materials & Processing Technology, College of Materials and Textile, Zhejiang Sci-Tech University, Hangzhou 310018 (China); Kundu, Subhas C. [Department of Biotechnology, Indian Institute of Technology (IIT) Kharagpur, West Bengal 721302 (India); Institute of Tissue Regeneration Engineering (ITREN), Dankook University, Cheonan 330-714 (Korea, Republic of); Yao, Juming, E-mail: yaoj@zstu.edu.cn [The Key Laboratory of Advanced Textile Materials and Manufacturing Technology of Ministry of Education, National Engineering Lab of Textile Fiber Materials & Processing Technology, College of Materials and Textile, Zhejiang Sci-Tech University, Hangzhou 310018 (China)

2016-09-15

To improve the fixations of the implant and implant-bone integration after joint arthroplasty from locally preventing inflammation and promoting the bone regeneration, we design a multifunctional biomaterial consisting of recombinant human bone morphogenetic protein 2 (rhBMP-2) and antibiotic loaded nano-hydroxyapatite with an alginate/gelatin sticky gel. We investigate its role for the prevention of the inflammation and possibility of inducing a new bone growth along with its adhesive ability. The stickiness exists in the composite, which may help to fix itself on the bone fracture surface. The composite sustains the antibacterial effect and promotes the proliferation and differentiation of MG63 cells in vitro. In vivo experimentation also shows that the composite gel has a role for the reduction of inflammation. It enhances the formation of new bone and blood vessels compared to both the sole rhBMP-2 and non-rhBMP-2/antibiotic loaded composite gels. The multifunctional composite provides a promising material for the prosthetic and bone tissue regeneration. - Highlights: • Multifunctional nanohydroxyapatite composite is fabricated. • The composite consists of nHAP, growth factor, antibiotic and alginate/gelatin gel. • The composite shows antibacterial effect and good cytocompatibility. • No adverse effect to the cells tested in vitro and in vivo.

Electrospun polycaprolactone/gelatin composites with enhanced cell–matrix interactions as blood vessel endothelial layer scaffolds

Energy Technology Data Exchange (ETDEWEB)

Jiang, Yong-Chao [National Center for International Research of Micro-Nano Molding Technology, Zhengzhou University, Zhengzhou (China); School of Mechanics and Engineering Science, Zhengzhou University, Zhengzhou (China); Department of Mechanical Engineering, University of Wisconsin-Madison, WI (United States); Wisconsin Institute for Discovery, University of Wisconsin-Madison, WI (United States); Jiang, Lin [National Center for International Research of Micro-Nano Molding Technology, Zhengzhou University, Zhengzhou (China); Department of Mechanical Engineering, University of Wisconsin-Madison, WI (United States); Wisconsin Institute for Discovery, University of Wisconsin-Madison, WI (United States); Huang, An [South China University of Technology, Guangzhou (China); Department of Mechanical Engineering, University of Wisconsin-Madison, WI (United States); Wisconsin Institute for Discovery, University of Wisconsin-Madison, WI (United States); Wang, Xiao-Feng [National Center for International Research of Micro-Nano Molding Technology, Zhengzhou University, Zhengzhou (China); School of Mechanics and Engineering Science, Zhengzhou University, Zhengzhou (China); Li, Qian [National Center for International Research of Micro-Nano Molding Technology, Zhengzhou University, Zhengzhou (China); Turng, Lih-Sheng, E-mail: turng@engr.wisc.edu [Department of Mechanical Engineering, University of Wisconsin-Madison, WI (United States); Wisconsin Institute for Discovery, University of Wisconsin-Madison, WI (United States)

2017-02-01

During the fabrication of tissue engineering scaffolds and subsequent tissue regeneration, surface bioactivity is vital for cell adhesion, spreading, and proliferation, especially for endothelium dysfunction repair. In this paper, synthetic polymer polycaprolactone (PCL) was blended with natural polymer gelatin at four different weight ratios followed by crosslinking (i.e., 100:0, 70:30, 50:50, 30:70, labeled as PCL-C, P7G3-C, P5G5-C, and P3G7-C) to impart enhanced bioactivity and tunable mechanical properties. The PCL/gelatin blends were first dissolved in 2,2,2-trifluroethanol (TFE) and supplementary acetic acid (1% relative to TFE) solvent, electrospun, and then cross-linked to produce PBS-proof fibrous scaffolds. Scanning electron micrographs (SEM) indicated that fibers of each sample were smooth and homogeneous, with the fiber diameters increasing from 1.01 ± 0.51 μm to 1.61 ± 0.46 μm as the content of gelatin increased. While thermal resistance and crystallization of the blends were affected by the presence of gelatin, as reflected by differential scanning calorimetry (DSC) results, water contact angle (WCA) tests confirmed that the scaffold surfaces became more hydrophilic. Tensile tests showed that PCL-C and P7G3-C scaffolds had mechanical properties comparable to those of human coronary arteries. As for cytocompatibility, skeleton staining images showed that human mesenchymal stem cells (hMSCs) had more favorable binding sites on PCL/gelatin scaffolds than those on PCL scaffolds. Cell proliferation assays revealed that P7G3-C scaffolds could support the most number of hMSCs. The results of this study demonstrated the enhanced cell-matrix interactions and potential use of electrospun PCL/gelatin scaffolds in the tissue engineering field, especially in wound dressings and endothelium regeneration. - Highlights: • Aqueous solution-resistant PCL/gelatin scaffolds were made via electrospinning. • PCL/gelatin composite scaffolds have tunable biophysical

Electrospun polycaprolactone/gelatin composites with enhanced cell–matrix interactions as blood vessel endothelial layer scaffolds

International Nuclear Information System (INIS)

Jiang, Yong-Chao; Jiang, Lin; Huang, An; Wang, Xiao-Feng; Li, Qian; Turng, Lih-Sheng

2017-01-01

During the fabrication of tissue engineering scaffolds and subsequent tissue regeneration, surface bioactivity is vital for cell adhesion, spreading, and proliferation, especially for endothelium dysfunction repair. In this paper, synthetic polymer polycaprolactone (PCL) was blended with natural polymer gelatin at four different weight ratios followed by crosslinking (i.e., 100:0, 70:30, 50:50, 30:70, labeled as PCL-C, P7G3-C, P5G5-C, and P3G7-C) to impart enhanced bioactivity and tunable mechanical properties. The PCL/gelatin blends were first dissolved in 2,2,2-trifluroethanol (TFE) and supplementary acetic acid (1% relative to TFE) solvent, electrospun, and then cross-linked to produce PBS-proof fibrous scaffolds. Scanning electron micrographs (SEM) indicated that fibers of each sample were smooth and homogeneous, with the fiber diameters increasing from 1.01 ± 0.51 μm to 1.61 ± 0.46 μm as the content of gelatin increased. While thermal resistance and crystallization of the blends were affected by the presence of gelatin, as reflected by differential scanning calorimetry (DSC) results, water contact angle (WCA) tests confirmed that the scaffold surfaces became more hydrophilic. Tensile tests showed that PCL-C and P7G3-C scaffolds had mechanical properties comparable to those of human coronary arteries. As for cytocompatibility, skeleton staining images showed that human mesenchymal stem cells (hMSCs) had more favorable binding sites on PCL/gelatin scaffolds than those on PCL scaffolds. Cell proliferation assays revealed that P7G3-C scaffolds could support the most number of hMSCs. The results of this study demonstrated the enhanced cell-matrix interactions and potential use of electrospun PCL/gelatin scaffolds in the tissue engineering field, especially in wound dressings and endothelium regeneration. - Highlights: • Aqueous solution-resistant PCL/gelatin scaffolds were made via electrospinning. • PCL/gelatin composite scaffolds have tunable biophysical

The effect of a chitosan-gelatin matrix and dexamethasone on the behavior of rabbit mesenchymal stem cells

International Nuclear Information System (INIS)

Medrado, G C B; Machado, C B; Valerio, P; Sanches, M D; Goes, A M

2006-01-01

Cartilage tissue has poor capability of self-repair, especially in the case of severe cartilage damage due to trauma or age-related degeneration. Cell-based tissue engineering using scaffolds has provided an option for the repair of defects in adult cartilage tissue. Mesenchymal stem cells (MSC) and chondrocytes are the two major cell sources for cartilage tissue engineering. The present study combined culture conditions of MSC in a chitosan-gelatin matrix in chondrogenic media to evaluate their effects on MSC viability and chondrogenesis for cartilage tissue engineering. MSC were harvested from rabbit bone marrows and cultured in chondrogenic media supplemented, or not, with dexamethasone in a chitosan-gelatin film (C-GF). The association of C-GF and dexamethasone promoted significant increase in cell adhesivity, viability and proliferation when compared to MCS cultured in media without dexamethasone or C-GF. In addition, dexamethasone promoted increase in the collagen concentration of MSC cultures. A reduction of alkaline phosphatase activity after three weeks of culture in chondrogenic media was verified. No influence of the C-GF or of dexamethasone was observed in this matter. Therefore, it is reasonable to suggest that biomaterial-based chitosan-gelatin and chondrogenic media supplemented with dexamethasone may stimulate the proliferation and differentiation of MSC according to the complex environmental conditions. The information presented here should be useful for the development of biomaterials to regulate the chondrogenesis of MSC suitable for cartilage tissue engineering

The effect of a chitosan-gelatin matrix and dexamethasone on the behavior of rabbit mesenchymal stem cells

Energy Technology Data Exchange (ETDEWEB)

Medrado, G C B [Medicine School, Federal University of Minas Gerais (UFMG), Belo Horizonte, MG (Brazil); Machado, C B [Biochemistry and Immunology Department, Biological Sciences Institute, UFMG - Federal University of Minas Gerais, mailbox 486, zip code 31270-901, Belo Horizonte, MG (Brazil); Valerio, P [Biochemistry and Immunology Department, Biological Sciences Institute, UFMG - Federal University of Minas Gerais, mailbox 486, zip code 31270-901, Belo Horizonte, MG (Brazil); Sanches, M D [Medicine School, Federal University of Minas Gerais (UFMG), Belo Horizonte, MG (Brazil); Goes, A M [Biochemistry and Immunology Department, Biological Sciences Institute, UFMG - Federal University of Minas Gerais, mailbox 486, zip code 31270-901, Belo Horizonte, MG (Brazil)

2006-09-15

Cartilage tissue has poor capability of self-repair, especially in the case of severe cartilage damage due to trauma or age-related degeneration. Cell-based tissue engineering using scaffolds has provided an option for the repair of defects in adult cartilage tissue. Mesenchymal stem cells (MSC) and chondrocytes are the two major cell sources for cartilage tissue engineering. The present study combined culture conditions of MSC in a chitosan-gelatin matrix in chondrogenic media to evaluate their effects on MSC viability and chondrogenesis for cartilage tissue engineering. MSC were harvested from rabbit bone marrows and cultured in chondrogenic media supplemented, or not, with dexamethasone in a chitosan-gelatin film (C-GF). The association of C-GF and dexamethasone promoted significant increase in cell adhesivity, viability and proliferation when compared to MCS cultured in media without dexamethasone or C-GF. In addition, dexamethasone promoted increase in the collagen concentration of MSC cultures. A reduction of alkaline phosphatase activity after three weeks of culture in chondrogenic media was verified. No influence of the C-GF or of dexamethasone was observed in this matter. Therefore, it is reasonable to suggest that biomaterial-based chitosan-gelatin and chondrogenic media supplemented with dexamethasone may stimulate the proliferation and differentiation of MSC according to the complex environmental conditions. The information presented here should be useful for the development of biomaterials to regulate the chondrogenesis of MSC suitable for cartilage tissue engineering.

Fabrication of gelatin-siloxane fibrous mats via sol-gel and electrospinning procedure and its application for bone tissue engineering

International Nuclear Information System (INIS)

Ren Lei; Wang Jun; Yang Fangyu; Wang Lin; Wang Dong; Wang Tianxiao; Tian Miaomiao

2010-01-01

Our strategy is to design and fabricate biomimetic and bioactive scaffolds that resemble the native extracellular matrix as closely as possible so as to create conducive living milieu that will induce cell to function naturally. In the present study, gelatin/siloxane (GS) hybrids were prepared by a sol-gel processing, and electrospinning technique was used to fabricate GS fibrous mats to support the growth of bone marrow-derived mesenchymal stem cells (BMSCs) for tissue engineering of bone. The results indicate that the porous structure and fiber size of the GS fibrous mats can be fine tuned by varying the viscosity of GS precursor solution. Additionally, the Ca 2+ -containing GS fibrous mats biomimetically deposited apatite in a simulated body fluid (SBF), as well as stimulating its BMSCs proliferation and differentiation in vitro, thereby dignifying its in vitro bioactivity.

Effects of ultraviolet irradiation on bonding strength between Co-Cr alloy and citric acid-crosslinked gelatin matrix.

Science.gov (United States)

Inoue, Motoki; Sasaki, Makoto; Katada, Yasuyuki; Taguchi, Tetsushi

2014-02-01

Novel techniques for creating a strong bond between polymeric matrices and biometals are required. We immobilized polymeric matrices on the surface of biometal for drug-eluting stents through covalent bond. We performed to improve the bonding strength between a cobalt-chromium alloy and a citric acid-crosslinked gelatin matrix by ultraviolet irradiation on the surface of cobalt-chromium alloy. The ultraviolet irradiation effectively generated hydroxyl groups on the surface of the alloy. The bonding strength between the gelatin matrix and the alloy before ultraviolet irradiation was 0.38 ± 0.02 MPa, whereas it increased to 0.48 ± 0.02 MPa after ultraviolet irradiation. Surface analysis showed that the citric acid derivatives occurred on the surface of the cobalt-chromium alloy through ester bond. Therefore, ester bond formation between the citric acid derivatives active esters and the hydroxyl groups on the cobalt-chromium alloy contributed to the enhanced bonding strength. Ultraviolet irradiation and subsequent immobilization of a gelatin matrix using citric acid derivatives is thus an effective way to functionalize biometal surfaces.

Enzyme-Gelatin Electrochemical Biosensors: Scaling Down

Directory of Open Access Journals (Sweden)

Hendrik A. Heering

2012-03-01

Full Text Available In this article we investigate the possibility of scaling down enzyme-gelatin modified electrodes by spin coating the enzyme-gelatin layer. Special attention is given to the electrochemical behavior of the selected enzymes inside the gelatin matrix. A glassy carbon electrode was used as a substrate to immobilize, in the first instance, horse heart cytochrome c (HHC in a gelatin matrix. Both a drop dried and a spin coated layer was prepared. On scaling down, a transition from diffusion controlled reactions towards adsorption controlled reactions is observed. Compared to a drop dried electrode, a spin coated electrode showed a more stable electrochemical behavior. Next to HHC, we also incorporated catalase in a spin coated gelatin matrix immobilized on a glassy carbon electrode. By spincoating, highly uniform sub micrometer layers of biocompatible matrices can be constructed. A full electrochemical study and characterization of the modified surfaces has been carried out. It was clear that in the case of catalase, gluteraldehyde addition was needed to prevent leaking of the catalase from the gelatin matrix.

Bioactivation of calcium deficient hydroxyapatite with foamed gelatin gel. A new injectable self-setting bone analogue.

Science.gov (United States)

Dessì, M; Alvarez-Perez, M A; De Santis, R; Ginebra, M P; Planell, J A; Ambrosio, L

2014-02-01

An alternative approach to bone repair for less invasive surgical techniques, involves the development of biomaterials directly injectable into the injury sites and able to replicate a spatially organized platform with features of bone tissue. Here, the preparation and characterization of an innovative injectable bone analogue made of calcium deficient hydroxyapatite and foamed gelatin is presented. The biopolymer features and the cement self-setting reaction were investigated by rheological analysis. The porous architecture, the evolution of surface morphology and the grains dimension were analyzed with electron microscopy (SEM/ESEM/TEM). The physico-chemical properties were characterized by X-ray diffraction and FTIR analysis. Moreover, an injection test was carried out to prove the positive effect of gelatin on the flow ensuing that cement is fully injectable. The cement mechanical properties are adequate to function as temporary substrate for bone tissue regeneration. Furthermore, MG63 cells and bone marrow-derived human mesenchymal stem cells (hMSCs) were able to migrate and proliferate inside the pores, and hMSCs differentiated to the osteoblastic phenotype. The results are paving the way for an injectable bone substitute with properties that mimic natural bone tissue allowing the successful use as bone filler for craniofacial and orthopedic reconstructions in regenerative medicine.

Electrospun gelatin/poly(ε-caprolactone) fibrous scaffold modified with calcium phosphate for bone tissue engineering

Energy Technology Data Exchange (ETDEWEB)

Rajzer, Izabella, E-mail: irajzer@ath.bielsko.pl [University of Bielsko-Biala (ATH), Department of Mechanical Engineering Fundamentals, Division of Materials Engineering, Willowa 2 Street, 43-309 Bielsko-Biała (Poland); Menaszek, Elżbieta [Jagiellonian University (UJ), Collegium Medicum, Department of Cytobiology, Medyczna 9 Street, 30-068 Cracow (Poland); Kwiatkowski, Ryszard [University of Bielsko-Biala (ATH), Faculty of Materials and Environmental Sciences, Institute of Textile Engineering and Polymer Materials, Willowa 2 Street, 43-309 Bielsko-Biała (Poland); Planell, Josep A.; Castano, Oscar [Institute for Bioengineering of Catalonia (IBEC), Biomaterials for Regenerative Therapies, Baldiri Reixac 15-21, 08028 Barcelona (Spain); Polytechnic University of Catalonia (UPC), Diagonal 647, 08028 Barcelona (Spain); CIBER-BBN The Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine, Barcelona (Spain)

2014-11-01

In this study gelatin (Gel) modified with calcium phosphate nanoparticles (SG5) and polycaprolactone (PCL) were used to prepare a 3D bi-layer scaffold by collecting electrospun PCL and gelatin/SG5 fibers separately in the same collector. The objective of this study was to combine the desired properties of PCL and Gel/SG5 in the same scaffold in order to enhance mineralization, thus improving the ability of the scaffold to bond to the bone tissue. The scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR) and the wide angle X-ray diffraction (WAXD) measurements confirmed that SG5 nanoparticles were successfully incorporated into the fibrous gelatin matrix. The composite Gel/SG5/PCL scaffold exhibited more enhanced mechanical properties than individual Gel and Gel/SG5 scaffolds. The presence of SG5 nanoparticles accelerated the nucleation and growth of apatite crystals on the surface of the composite Gel/SG5/PCL scaffold in simulated body fluid (SBF). The osteoblast response in vitro to developed electrospun scaffolds (PCL and Gel/SG5/PCL) was investigated by using normal human primary NHOst cell lines. NHOst cell culture studies showed that higher alkaline phosphatase (ALP) activity and better mineralization were obtained in the case of composite materials than in pure PCL scaffolds. The mechanically strong PCL scaffold served as a skeleton, while the Gel/SG5 fibers facilitated cell spreading and mineralization of the scaffold. - Highlights: • Bi-layer scaffolds were produced by electrospinning method. • The addition of nanoparticles enhanced the bioactivity of scaffold. • Bi-layer scaffold enhanced ALP activity and NHOst cell mineralization.

Sequentially-crosslinked biomimetic bioactive glass/gelatin methacryloyl composites hydrogels for bone regeneration.

Science.gov (United States)

Zheng, Jiafu; Zhao, Fujian; Zhang, Wen; Mo, Yunfei; Zeng, Lei; Li, Xian; Chen, Xiaofeng

2018-08-01

In recent years, gelatin-based composites hydrogels have been intensively investigated because of their inherent bioactivity, biocompatibility and biodegradability. Herein, we fabricated photocrosslinkable biomimetic composites hydrogels from bioactive glass (BG) and gelatin methacryloyl (GelMA) by a sequential physical and chemical crosslinking (gelation + UV) approach. The results showed that the compressive modulus of composites hydrogels increased significantly through the sequential crosslinking approach. The addition of BG resulted in a significant increase in physiological stability and apatite-forming ability. In vitro data indicated that BG/GelMA composites hydrogels promoted cell attachment, proliferation and differentiation. Overall, the BG/GelMA composites hydrogels combined the advantages of good biocompatibility and bioactivity, and had potential applications in bone regeneration. Copyright © 2018. Published by Elsevier B.V.

Study of the effects of heat-treatment of hydroxyapatite synthesized in gelatin matrix

Science.gov (United States)

Zaits, A. V.; Golovanova, O. A.; Kuimova, M. V.

2017-01-01

In the study, the isothermal thermogravimetric analysis (TGA) of hydroxyapatite synthesized in gelatin matrix (HAG) has been performed. 3 wt.% HAG samples were synthesized from the solution simulating the human extracellular fluid (SBF). X-ray diffraction and IR spectroscopy were used to determine the composition. During the experiment, increase in the calcination temperature up to 200°C-800°C was found to cause weight loss. The study of phase composition revealed that heat treatment does not affect the phase composition of the solid phase, which is composed of hydroxylapatite (HA). The prepared HAG (3 wt.% gelatin) samples are shown to have low thermal stability; the degradation of the samples occurs at 400°C.

Barium-cross-linked alginate-gelatine microcapsule as a potential platform for stem cell production and modular tissue formation.

Science.gov (United States)

Alizadeh Sardroud, Hamed; Nemati, Sorour; Baradar Khoshfetrat, Ali; Nabavinia, Mahbobeh; Beygi Khosrowshahi, Younes

2017-08-01

Influence of gelatine concentration and cross-linker ions of Ca 2+ and Ba 2+ was evaluated on characteristics of alginate hydrogels and proliferation behaviours of model adherent and suspendable stem cells of fibroblast and U937 embedded in alginate microcapsules. Increasing gelatine concentration to 2.5% increased extent of swelling to 15% and 25% for barium- and calcium-cross-linked hydrogels, respectively. Mechanical properties also decreased with increasing swelling of hydrogels. Both by increasing gelatine concentration and using barium ions increased considerably the proliferation of encapsulated model stem cells. Barium-cross-linked alginate-gelatine microcapsule tested for bone building block showed a 13.5 ± 1.5-fold expansion for osteoblast cells after 21 days with deposition of bone matrix. The haematopoietic stem cells cultured in the microcapsule after 7 days also showed up to 2-fold increase without adding any growth factor. The study demonstrates that barium-cross-linked alginate-gelatine microcapsule has potential for use as a simple and efficient 3D platform for stem cell production and modular tissue formation.

Radiation synthesis of gelatin/CM-chitosan/{beta}-tricalcium phosphate composite scaffold for bone tissue engineering

Energy Technology Data Exchange (ETDEWEB)

Zhou Ying [College of Engineering, Peking University, Beijing 100871 (China); Center for Biomedical Materials and Tissue Engineering, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100871 (China); Xu Ling, E-mail: lingxu@pku.edu.cn [College of Engineering, Peking University, Beijing 100871 (China); Center for Biomedical Materials and Tissue Engineering, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100871 (China); Zhang Xiangmei; Zhao Yinghui [Center for Biomedical Materials and Tissue Engineering, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100871 (China); Wei Shicheng, E-mail: sc-wei@pku.edu.cn [Center for Biomedical Materials and Tissue Engineering, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100871 (China); Department of Oral and Maxillofacial Surgery, School and Hospital of Stomatology, Peking University, Beijing 100081 (China); Zhai Maolin [Beijing National Laboratory for Molecular Sciences, Department of Applied Chemistry, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871 (China)

2012-05-01

A series of biodegradable composite scaffolds was fabricated from an aqueous solution of gelatin, carboxymethyl chitosan (CM-chitosan) and {beta}-tricalcium phosphate ({beta}-TCP) by radiation-induced crosslinking at ambient temperature. Ultrasonic treatment on the polymer solutions significantly influenced the distribution of {beta}-TCP particles. An ultrasonic time of 20 min, followed by 30 kGy irradiation induced a crosslinked scaffold with homogeneous distribution of {beta}-TCP particles, interconnected porous structure, sound swelling capacity and mechanical strength. Fourier Transform Infrared Spectroscopy and X-ray Diffraction analysis indicated that {beta}-TCP successfully incorporated with the network of gelatin and CM-chitosan. In vivo implantation of the scaffold into the mandible of beagle dog revealed that the scaffolds had excellent biocompatibility and the presence of {beta}-TCP can accelerate bone regeneration. The comprehensive results of this study paved way for the application of gelatin/CM-chitosan/{beta}-TCP composite scaffolds as candidate of bone tissue engineering material. - Highlights: Black-Right-Pointing-Pointer Radiation induced a crosslinked scaffold with interconnected porous structure. Black-Right-Pointing-Pointer Ultrasonic time of 20 min led to homogenerously distribution of {beta}-TCP. Black-Right-Pointing-Pointer Increasing amount of {beta}-TCP would restrict the swelling properties. Black-Right-Pointing-Pointer Proper fraction of {beta}-TCP will promote the mechanical properties of the scaffolds. Black-Right-Pointing-Pointer Hybrid of {beta}-TCP promoted the bone regeneration of the mandibles of beagle dogs.

Design, synthesis, and initial evaluation of D-glyceraldehyde crosslinked gelatin-hydroxyapatite as a potential bone graft substitute material

Science.gov (United States)

Florschutz, Anthony Vatroslav

Utilization of bone grafts for the treatment of skeletal pathology is a common practice in orthopaedic, craniomaxillofacial, dental, and plastic surgery. Autogenous bone graft is the established archetype but has disadvantages including donor site morbidity, limited supply, and prolonging operative time. In order to avoid these and other issues, bone graft substitute materials are becoming increasingly prevalent among surgeons for reconstructing skeletal defects and arthrodesis applications. Bone graft substitutes are biomaterials, biologics, and guided tissue/bone regenerative devices that can be used alone or in combinations as supplements or alternatives to autogenous bone graft. There is a growing interest and trend to specialize graft substitutes for specific indications and although there is good rationale for this indication-specific approach, the development and utility of a more universal bone graft substitute may provide a better answer for patients and surgeons. The aim of the present research focuses on the design, synthesis, and initial evaluation of D-glyceraldehyde crosslinked gelatin-hydroxyapatite composites for potential use as a bone graft substitutes. After initial establishment of rational material design, gelatinhydroxyapatite scaffolds were fabricated with different gelatin:hydroxyapatite ratios and crosslinking concentrations. The synthesized scaffolds were subsequently evaluated on the basis of their swelling behavior, porosity, density, percent composition, mechanical properties, and morphology and further assessed with respect to cell-biomaterial interaction and biomineralization in vitro. Although none of the materials achieved mechanical properties suitable for structural graft applications, a reproducible material design and synthesis was achieved with properties recognized to facilitate bone formation. Select scaffold formulations as well as a subset of scaffolds loaded with recombinant human bone morphogenetic protein-2 were

D-Glucose as a modifying agent in gelatin/collagen matrix and reservoir nanoparticles for Calendula officinalis delivery.

Science.gov (United States)

Lam, P-L; Kok, S H-L; Bian, Z-X; Lam, K-H; Tang, J C-O; Lee, K K-H; Gambari, R; Chui, C-H

2014-05-01

Gelatin/Collagen-based matrix and reservoir nanoparticles require crosslinkers to stabilize the formed nanosuspensions, considering that physical instability is the main challenge of nanoparticulate systems. The use of crosslinkers improves the physical integrity of nanoformulations under the-host environment. Aldehyde-based fixatives, such as formaldehyde and glutaraldehyde, have been widely applied to the crosslinking process of polymeric nanoparticles. However, their potential toxicity towards human beings has been demonstrated in many previous studies. In order to tackle this problem, D-glucose was used during nanoparticle formation to stabilize the gelatin/collagen-based matrix wall and reservoir wall for the deliveries of Calendula officinalis powder and oil, respectively. In addition, therapeutic selectivity between malignant and normal cells could be observed. The C. officinalis powder loaded nanoparticles significantly strengthened the anti-cancer effect towards human breast adenocarcinoma MCF7 cells and human hepatoma SKHep1 cells when compared with the free powder. On the contrary, the nanoparticles did not show significant cytotoxicity towards normal esophageal epithelial NE3 cells and human skin keratinocyte HaCaT cells. On the basis of these evidences, D-glucose modified gelatin/collagen matrix nanoparticles containing C. officinalis powder might be proposed as a safer alternative vehicle for anti-cancer treatments. Copyright © 2014 Elsevier B.V. All rights reserved.

Fish gelatin.

Science.gov (United States)

Boran, Gokhan; Regenstein, Joe M

2010-01-01

Gelatin is a multifunctional ingredient used in foods, pharmaceuticals, cosmetics, and photographic films as a gelling agent, stabilizer, thickener, emulsifier, and film former. As a thermoreversible hydrocolloid with a narrower gap between its melting and gelling temperatures, both of which are below human body temperature, gelatin provides unique advantages over carbohydrate-based gelling agents. Gelatin is mostly produced from pig skin, and cattle hides and bones. Some alternative raw materials have recently gained attention from both researchers and the industry not just because they overcome religious concerns shared by Jews and Muslims but also because they provide, in some cases, technological advantages over mammalian gelatins. Fish skins from a number of fish species are among the other sources that have been comprehensively studied as sources for gelatin production. Fish skins have a significant potential for the production of high-quality gelatin with different melting and gelling temperatures over a much wider range than mammalian gelatins, yet still have a sufficiently high gel strength and viscosity. Gelatin quality is industrially determined by gel strength, viscosity, melting or gelling temperatures, the water content, and microbiological safety. For gelatin manufacturers, yield from a particular raw material is also important. Recent experimental studies have shown that these quality parameters vary greatly depending on the biochemical characteristics of the raw materials, the manufacturing processes applied, and the experimental settings used for quality control tests. In this review, the gelatin quality achieved from different fish species is reviewed along with the experimental procedures used to determine gelatin quality. In addition, the chemical structure of collagen and gelatin, the collagen-gelatin conversion, the gelation process, and the gelatin market are discussed. Copyright © 2010 Elsevier Inc. All rights reserved.

Three-Dimensional Supermacroporous Carrageenan-Gelatin Cryogel Matrix for Tissue Engineering Applications

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Archana Sharma

2013-01-01

Full Text Available A tissue-engineered polymeric scaffold should provide suitable macroporous structure similar to that of extracellular matrix which can induce cellular activities and guide tissue regeneration. Cryogelation is a technique in which appropriate monomers or polymeric precursors frozen at sub-zero temperature leads to the formation of supermacroporous cryogel matrices. In this study carrageenan-gelatin (natural polymers cryogels were synthesized by using glutaraldehyde and 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride and N-hydroxysuccinimide (EDC-NHS as crosslinking agent at optimum concentrations. Matrices showed large and interconnected pores which were in the range of 60–100 μm diameter. Unconfined compression analysis showed elasticity and physical integrity of all cryogels, as these matrices regained their original length after 90% compressing from the original size. Moreover Young’s modulus was found to be in the range of 4–11 kPa for the dry cryogel sections. These cryogels also exhibited good in vitro degradation capacity at 37 °C within 4 weeks of incubation. Supermacroporous carrageenan-gelatin cryogels showed efficient cell adherence and proliferation of Cos-7 cells which was examined by SEM. PI nuclear stain was used to observe cell-matrix interaction. Cytotoxicity of the scaffolds was checked by MTT assay which showed that cryogels are biocompatible and act as a potential material for tissue engineering and regenerative medicine.

Enzymatically crosslinked carboxymethyl-chitosan/gelatin/nano-hydroxyapatite injectable gels for in situ bone tissue engineering application

Energy Technology Data Exchange (ETDEWEB)

Mishra, Debasish; Bhunia, Bibhas; Banerjee, Indranil [Department of Biotechnology, Indian Institute of Technology Kharagpur (India); Datta, Pallab; Dhara, Santanu [School of Medical Science and Technology, Indian Institute of Technology Kharagpur (India); Maiti, Tapas K., E-mail: maititapask@gmail.com [Department of Biotechnology, Indian Institute of Technology Kharagpur (India)

2011-10-10

Present study reports synthesis and characterization of an enzymatically crosslinked injectable gel (iGel) suitable for cell based bone tissue engineering application. The gel comprises of carboxymethyl-chitosan (CMC)/gelatin/nano-hydroxyapatite (nHAp) susceptible to tyrosinase/p-cresol mediated in situ gelling at physiological temperature. Study revealed that a combination of tyrosinase (60U) and p-cresol (2 mM) as crosslinking agents yield rigid gels at physiological temperature when applied to CMC/gelatin within 35 min in presence or absence of nHAp. Rheological study in conjugation with FT-IR analysis showed that an increase in CMC concentration in the gel leads to higher degree of crosslinking and higher strength. Scanning electron microscopy showed that pore sizes of iGels increased with higher gelatin concentration. In vitro study of osteoblast cell proliferation and differentiation showed that, although all iGels are supportive towards the growth of primary osteoblast cells, GC1:1 supported cellular differentiation to the maximum. Application of iGels in mice revealed that stability of the in situ formed gels depends on the degree of crosslinking and CMC concentration. In conclusion, the iGels may be used in treating irregular small bone defects with minimal clinical invasion as well as for bone cell delivery. - Research Highlights: {yields} Enzymatically crosslinked injectable gel made up of CM-chitosan (C)/gelatin (G)/nHAp. {yields} Tyrosinase/p-cresol used for crosslinking and in situ gelling of polymers at 37deg. C. {yields} 60U tyrosinase and 2mM p-cresol is needed for gelation in 35 min. {yields} Higher GC ratio manifests lower crosslinking and gel strength but higher porosity. {yields} GC1:1 shows maximum in vivo gel stability and in vitro osteoblast differentiation.

Bone morphogenetic protein-2 loaded poly(D,L-lactide-co-glycolide microspheres enhance osteogenic potential of gelatin/hydroxyapatite/β-tricalcium phosphate cryogel composite for alveolar ridge augmentation

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Hao-Chieh Chang

2017-12-01

Full Text Available Background/Purpose: Sufficient bony support is essential to ensure the success of dental implant osseointegration. However, the reconstruction of vertical ridge deficiencies is still a major challenge for dental implants. This study introduced a novel treatment strategy by infusing poly(D,L-lactide-co-glycolide (PLGA microspheres encapsulating bone morphogenetic protein-2 (BMP-2 within a gelatin/hydroxyapatite/β-tricalcium phosphate (gelatin/HA/β-TCP cryogel composite to facilitate supra-alveolar ridge augmentation. Methods: The gelatin scaffold was crosslinked using cryogel technique, and HA/β-TCP particles were mechanically entrapped to form the gelatin/HA/β-TCP composite. Co-axial electrohydrodynamic atomization technology was used to fabricate PLGA microspheres encapsulating BMP-2. The composites of gelatin/HA/β-TCP alone, with infusion of BMP-2 solution (BMPi or microspheres (BMPm, were fixed on rat mandibles using a titanium mini-implant for 4 weeks, and the therapeutic efficiency was evaluated by micro-computed tomography, bone fluorochrome, and histology. Results: The gelatin/HA/β-TCP composite was homogenously porous, and BMP-2 was sustained release from the microspheres without initial burst release. Ridge augmentation was noted in all specimens treated with the gelatin/HA/β-TCP composite, and greater bone deposition ratio were noted in Groups BMPi and BMPm. Compared with Group BMPi, specimens in Group BMPm showed significantly greater early osteogenesis and evident osseointegration in the supra-alveolar level. Conclusion: BMP-2 loaded PLGA microspheres effectively promoted osteogenic potential of the gelatin/HA/β-TCP composite and facilitated supra-alveolar ridge augmentation in vivo. Keywords: bone morphogenetic protein-2, bone regeneration, dental implant, tissue engineering, tissue scaffolds

Characterization of partially hydrolyzed OCP crystals deposited in a gelatin matrix as a scaffold for bone tissue engineering

Science.gov (United States)

Ezoe, Yushi; Anada, Takahisa; Yamazaki, Hajime; Handa, Takuto; Kobayashi, Kazuhito; Takahashi, Tetsu; Suzuki, Osamu

2015-03-01

The present study was designed to investigate how hydrolysis of octacalcium phosphate (OCP) into hydroxyapatite is affected by the presence of gelatin (Gel) molecules and how osteoblastic cells respond to the resultant OCP hydrolyzate/Gel composites as the hydrolysis advances. OCP was prepared from a solution containing calcium and phosphate ions and Gel molecules, having a composition to produce a 40 wt% OCP as a final co-precipitate as the OCP/Gel. The precipitate was further incubated up to 40 h to advance the hydrolysis of OCP. These precipitates were processed to mold OCP/Gel sponges through lyophilization and dehydrothermal treatment. Chemical analysis, X-ray diffraction, Fourier transform infrared spectroscopy, transmission electron microscopy, and selected area electron diffraction revealed that the hydrolysis of OCP/Gel composite in hot water advanced in a time-dependent manner and faster than hydrolysis of OCP alone. The effect of Gel on the OCP hydrolysis was further examined in the presence of distinct concentrations of Gel molecules in hot water, showing that the Gel enhanced the hydrolysis as the concentration increased. Proliferation and differentiation of mouse bone marrow stromal ST-2 cells on the hydrolyzed OCP/Gel composites were compatible with Gel sponge alone after 21 days of culture, suggesting that these composites could be a candidate as a scaffold in bone tissue engineering.

Characterization of partially hydrolyzed OCP crystals deposited in a gelatin matrix as a scaffold for bone tissue engineering

Energy Technology Data Exchange (ETDEWEB)

Ezoe, Yushi [Tohoku University Graduate School of Dentistry, Division of Oral and Maxillofacial Surgery (Japan); Anada, Takahisa [Tohoku University Graduate School of Dentistry, Division of Craniofacial Function Engineering (Japan); Yamazaki, Hajime [The Forsyth Institute, Department of Applied Oral Sciences, Center for Biomineralization (United States); Handa, Takuto; Kobayashi, Kazuhito; Takahashi, Tetsu [Tohoku University Graduate School of Dentistry, Division of Oral and Maxillofacial Surgery (Japan); Suzuki, Osamu, E-mail: suzuki-o@m.tohoku.ac.jp [Tohoku University Graduate School of Dentistry, Division of Craniofacial Function Engineering (Japan)

2015-03-15

The present study was designed to investigate how hydrolysis of octacalcium phosphate (OCP) into hydroxyapatite is affected by the presence of gelatin (Gel) molecules and how osteoblastic cells respond to the resultant OCP hydrolyzate/Gel composites as the hydrolysis advances. OCP was prepared from a solution containing calcium and phosphate ions and Gel molecules, having a composition to produce a 40 wt% OCP as a final co-precipitate as the OCP/Gel. The precipitate was further incubated up to 40 h to advance the hydrolysis of OCP. These precipitates were processed to mold OCP/Gel sponges through lyophilization and dehydrothermal treatment. Chemical analysis, X-ray diffraction, Fourier transform infrared spectroscopy, transmission electron microscopy, and selected area electron diffraction revealed that the hydrolysis of OCP/Gel composite in hot water advanced in a time-dependent manner and faster than hydrolysis of OCP alone. The effect of Gel on the OCP hydrolysis was further examined in the presence of distinct concentrations of Gel molecules in hot water, showing that the Gel enhanced the hydrolysis as the concentration increased. Proliferation and differentiation of mouse bone marrow stromal ST-2 cells on the hydrolyzed OCP/Gel composites were compatible with Gel sponge alone after 21 days of culture, suggesting that these composites could be a candidate as a scaffold in bone tissue engineering.

Heterotopic new bone formation causes resorption of the inductive bone matrix

International Nuclear Information System (INIS)

Nilsson, O.S.; Persson, P.E.; Ekelund, A.

1990-01-01

The bone matrix of growing rats was labeled by multiple injections of 3H-proline, and demineralized bone matrix (DBM) was prepared. The DBM was allotransplanted heterotopically into growing rats. New bone formation was induced in and around the implants. The new bone formation was accompanied by a decrease in the content of 3H; 20 and 30 days after implantation, 72% and 46%, respectively, of the activity remained in the implants. Daily injections of indomethacin (2 mg/kg) inhibited calcium uptake by about 20% at 20 and 30 days and inhibited the release of 3H from the DBM to a similar degree. Heterotopic bone induction by DBM is accompanied by matrix resorption, and inhibition of the new bone formation decreases the resorption of DBM

Fabrication and Physical Evaluation of Gelatin-Coated Carbonate Apatite Foam

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Kanae Hara

2016-08-01

Full Text Available Carbonate apatite (CO3Ap foam has gained much attention in recent years because of its ability to rapidly replace bone. However, its mechanical strength is extremely low for clinical use. In this study, to understand the potential of gelatin-reinforced CO3Ap foam for bone replacement, CO3Ap foam was reinforced with gelatin and the resulting physical characteristics were evaluated. The mechanical strength increased significantly with the gelatin reinforcement. The compressive strength of gelatin-free CO3Ap foam was 74 kPa whereas that of the gelatin-reinforced CO3Ap foam, fabricated using 30 mass % gelatin solution, was approximately 3 MPa. Heat treatment for crosslinking gelatin had little effect on the mechanical strength of the foam. The gelatin-reinforced foam did not maintain its shape when immersed in a saline solution as this promoted swelling of the gelatin; however, in the same conditions, the heat-treated gelatin-reinforced foam proved to be stable. It is concluded, therefore, that heat treatment is the key to the fabrication of stable gelatin-reinforced CO3Ap foam.

Extraskeletal and intraskeletal new bone formation induced by demineralized bone matrix combined with bone marrow cells

International Nuclear Information System (INIS)

Lindholm, T.S.; Nilsson, O.S.; Lindholm, T.C.

1982-01-01

Dilutions of fresh autogenous bone marrow cells in combination with allogeneic demineralized cortical bone matrix were tested extraskeletally in rats using roentgenographic, histologic, and 45 Ca techniques. Suspensions of bone marrow cells (especially diluted 1:2 with culture media) combined with demineralized cortical bone seemed to induce significantly more new bone than did demineralized bone, bone marrow, or composite grafts with whole bone marrow, respectively. In a short-term spinal fusion experiment, demineralized cortical bone combined with fresh bone marrow produced new bone and bridged the interspace between the spinous processes faster than other transplantation procedures. The induction of undifferentiated host cells by demineralized bone matrix is further complemented by addition of autogenous, especially slightly diluted, bone marrow cells

Extraction of gelatin from catfish bone using NaOH and its utilization as a template on mesoporous silica alumina

Science.gov (United States)

Nuryanto, R.; Trisunaryanti, W.; Falah, I. I.; Triyono

2018-04-01

Gelatin extraction from catfish bone using NaOH and its utilization as a template on a synthesis of mesoporous silica-alumina had been investigated. The extraction was prepared by immersing 25 g catfish bone in 125 mL of NaOH in concentration of 0.0; 0.05; 0.10; 0.15 and 0.20 M for 24 h, then washing with demineralized water until pH 7, followed by immersed the bone into 125 mL of 1 M HCl for 1 h, then washed using demineralized water into pH 5. To produce gelatin the bone was refluxed with 100 mL demineralized water at 70°C for 5 h then evaporated at 50°C. The dry gelatin was characterized using FTIR and electrophoresis (SDS-PAGE). The best performance of gelatin was produced by NaOH 0.10 M. The gelatin consists of amide A, B, I, II, III and molecular weight of 25-200kDa. Silica and Alumina material prepared from Lapindo mud extraction. Dry Lapindo mud crushed and filtered until pass 100 mesh, then reflux using 6 M HCl (1:4 w/V) at 90°C for 5h then filtered. The filtrate was consisting alumina solution adding with 6 M NaOH (2/3 V/V) them filtered. The filtrate then injected by CO2 gas for 30 minutes and filtered, the residue was calcined at 500°C for 5h. The residual of Lapindo mud dried and refluxed with 6 M NaOH (1:4 w/v) at 90 °C. After 5h filtered and the filtrate added by HCl to pH 8 and filtered, the residual then dried. The Si and Al were then analyzed by XRF and consist of silica and alumina for 99.1 and 87.73%, respectively. Silica-alumina was prepared using silica and alumina extracted from Lapindo mud. 6 g of SiO2 and 2 g of NaOH was immersed in 62 mL of demineralized water then added with alumina solution (0.204 g alumina in 30 mL demineralized water). The gelatin solution (5 g gelatin in 70 mL demineralized water) was dropped into the silica-alumina while stirring at 50°C for 4 h and aging for 24 h. The synthesized silica alumina was analysed using FTIR and surface area analyser. The FT-IR spectra indicated the TO4 (T=Si, Al) vibration at wave

Characterization and bioactivity of nano-submicro octacalcium phosphate/gelatin composite

International Nuclear Information System (INIS)

Miura, Kei-ichiro; Anada, Takahisa; Honda, Yoshitomo; Shiwaku, Yukari; Kawai, Tadashi; Echigo, Seishi; Takahashi, Tetsu; Suzuki, Osamu

2013-01-01

The present study was designed to investigate the physicochemical and bioactive properties of a nano-submicro sized octacalcium phosphate (OCP)-dispersed gelatin (Gel) composite (nano-submicro OCP/Gel) used as a bone substitute material in various bone defects. Well-grown, synthesized OCP was mechanically ground from 100 to 300 μm-sieved granules to particles that were approximately 500 nm in size. Then, 50 wt% of the nano-submicro OCP was mixed with porcine skin-derived acid extracted gelatin. The mixture was molded and lyophilized and then subjected to dehydrothermal crosslinking. X-ray diffraction (XRD) and Fourier transform infrared (FTIR) spectroscopy showed that the structure of OCP was retained even after mechanical grinding to a nano-submicro scale level as well as inclusion in the Gel matrix. The bioactivity of nano-submicro OCP/Gel was examined by immersing the composite in simulated body fluid (SBF) for 7 days and by implanting it in rat critical-sized calvaria defects for 8 weeks. The nano-submicro OCP tended to convert to low crystalline hydroxyapatite (HA) in SBF as assessed by XRD. The nano-submicro OCP/Gel exhibited osteoconductivity in vivo, yielding new bone formation that was closely associated with the implanted composite. These results suggest that the nano-submicro OCP/Gel composite exhibits similar osteoconductivity as observed in other OCP-based materials previously reported and could be used as a bone substitute material for repairing various defects in bone.

Characterization and bioactivity of nano-submicro octacalcium phosphate/gelatin composite

Energy Technology Data Exchange (ETDEWEB)

Miura, Kei-ichiro [Division of Craniofacial Function Engineering, Tohoku University Graduate School of Dentistry, Sendai (Japan); Division of Oral and Maxillofacial Surgery, Tohoku University Graduate School of Dentistry, Sendai (Japan); Anada, Takahisa; Honda, Yoshitomo [Division of Craniofacial Function Engineering, Tohoku University Graduate School of Dentistry, Sendai (Japan); Shiwaku, Yukari [Division of Craniofacial Function Engineering, Tohoku University Graduate School of Dentistry, Sendai (Japan); Division of Advanced Prosthetic Dentistry, Tohoku University Graduate School of Dentistry, Sendai (Japan); Kawai, Tadashi; Echigo, Seishi; Takahashi, Tetsu [Division of Oral and Maxillofacial Surgery, Tohoku University Graduate School of Dentistry, Sendai (Japan); Suzuki, Osamu, E-mail: suzuki-o@m.tohoku.ac.jp [Division of Craniofacial Function Engineering, Tohoku University Graduate School of Dentistry, Sendai (Japan)

2013-10-01

The present study was designed to investigate the physicochemical and bioactive properties of a nano-submicro sized octacalcium phosphate (OCP)-dispersed gelatin (Gel) composite (nano-submicro OCP/Gel) used as a bone substitute material in various bone defects. Well-grown, synthesized OCP was mechanically ground from 100 to 300 μm-sieved granules to particles that were approximately 500 nm in size. Then, 50 wt% of the nano-submicro OCP was mixed with porcine skin-derived acid extracted gelatin. The mixture was molded and lyophilized and then subjected to dehydrothermal crosslinking. X-ray diffraction (XRD) and Fourier transform infrared (FTIR) spectroscopy showed that the structure of OCP was retained even after mechanical grinding to a nano-submicro scale level as well as inclusion in the Gel matrix. The bioactivity of nano-submicro OCP/Gel was examined by immersing the composite in simulated body fluid (SBF) for 7 days and by implanting it in rat critical-sized calvaria defects for 8 weeks. The nano-submicro OCP tended to convert to low crystalline hydroxyapatite (HA) in SBF as assessed by XRD. The nano-submicro OCP/Gel exhibited osteoconductivity in vivo, yielding new bone formation that was closely associated with the implanted composite. These results suggest that the nano-submicro OCP/Gel composite exhibits similar osteoconductivity as observed in other OCP-based materials previously reported and could be used as a bone substitute material for repairing various defects in bone.

A gelatin composite scaffold strengthened by drug-loaded halloysite nanotubes.

Science.gov (United States)

Ji, Lijun; Qiao, Wei; Zhang, Yuheng; Wu, Huayu; Miao, Shiyong; Cheng, Zhilin; Gong, Qianming; Liang, Ji; Zhu, Aiping

2017-09-01

Mechanical properties and anti-infection are two of the most concerned issues for artificial bone grafting materials. Bone regeneration porous scaffolds with sustained drug release were developed by freeze-drying the mixture of nanosized drug-loaded halloysite nanotubes (HNTs) and gelatin. The scaffolds showed porous structure and excellent biocompatibility. The mechanical properties of the obtained composite scaffolds were enhanced significantly by HNTs to >300%, comparing to those of gelatin scaffold, and match to those of natural cancellous bones. The ibuprofen-loaded HNTs incorporated in the scaffolds allowed extended drug release over 100h, comparing to 8h when directly mixed the drug into the gelatin scaffold. The biological properties of the composite scaffolds were investigated by culturing MG63 cells on them. The HNTs/gelatin scaffolds with excellent mechanical properties and sustained drug release could be a promising artificial bone grating material. Copyright © 2017 Elsevier B.V. All rights reserved.

Preparation and Evaluation of Gelatin-Chitosan-Nanobioglass 3D Porous Scaffold for Bone Tissue Engineering

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Kanchan Maji

2016-01-01

Full Text Available The aim of the present study was to prepare and characterize bioglass-natural biopolymer based composite scaffold and evaluate its bone regeneration ability. Bioactive glass nanoparticles (58S in the size range of 20–30 nm were synthesized using sol-gel method. Porous scaffolds with varying bioglass composition from 10 to 30 wt% in chitosan, gelatin matrix were fabricated using the method of freeze drying of its slurry at 40 wt% solids loading. Samples were cross-linked with glutaraldehyde to obtain interconnected porous 3D microstructure with improved mechanical strength. The prepared scaffolds exhibited >80% porosity with a mean pore size range between 100 and 300 microns. Scaffold containing 30 wt% bioglass (GCB 30 showed a maximum compressive strength of 2.2±0.1 MPa. Swelling and degradation studies showed that the scaffold had excellent properties of hydrophilicity and biodegradability. GCB 30 scaffold was shown to be noncytotoxic and supported mesenchymal stem cell attachment, proliferation, and differentiation as indicated by MTT assay and RUNX-2 expression. Higher cellular activity was observed in GCB 30 scaffold as compared to GCB 0 scaffold suggesting the fact that 58S bioglass nanoparticles addition into the scaffold promoted better cell adhesion, proliferation, and differentiation. Thus, the study showed that the developed composite scaffolds are potential candidates for regenerating damaged bone tissue.

Fabrication and in vitro evaluation of a sponge-like bioactive-glass/gelatin composite scaffold for bone tissue engineering

International Nuclear Information System (INIS)

Nadeem, Danish; Kiamehr, Mostafa; Yang, Xuebin; Su, Bo

2013-01-01

In this work a bioactive composite scaffold, comprised of bioactive-glass and gelatin, is introduced. Through direct foaming a sponge-like composite of a sol–gel derived bioactive-glass (70S30C; 70% SiO 2 , 30% CaO) and porcine gelatin was developed for use as a biodegradable scaffold for bone tissue engineering. The composite was developed to provide a suitable alternative to synthetic polymer based scaffolds, allowing directed regeneration of bone tissue. The fabricated scaffold was characterised through X-ray microtomography, scanning electron and light microscopy demonstrating a three dimensionally porous and interconnected structure, with an average pore size (170 μm) suitable for successful cell proliferation and tissue ingrowth. Acellular bioactivity was assessed through apatite formation during submersion in simulated body fluid (SBF) whereby the rate and onset of apatite nucleation was found to be comparable to that of bioactive-glass. Modification of dehydrothermal treatment parameters induced varying degrees of crosslinking, allowing the degradation of the composite to be tailored to suit specific applications and establishing its potential for a wide range of applications. Use of genipin to supplement crosslinking by dehydrothermal treatment provided further means of modifying degradability. Biocompatibility of the composite was qualified through successful cultures of human dental pulp stem cells (HDPSCs) on samples of the composite scaffold. Osteogenic differentiation of HDPSCs and extracellular matrix deposition were confirmed through positive alkaline phosphatase staining and immunohistochemistry. - Highlights: ► Optimised composition and fabrication produced sponge-like porosity (pore size ∼ 170 μm). ► Maximum aqueous stability via dehydrothermal treatment at 145 °C, for 48 h ► Biocompatibility and osteogenic potential confirmed via successful HDPSC cultures. ► Minimal toxicity exhibited in optimally crosslinked samples (10 m

Fabrication and in vitro evaluation of a sponge-like bioactive-glass/gelatin composite scaffold for bone tissue engineering

Energy Technology Data Exchange (ETDEWEB)

Nadeem, Danish [Biomaterials Engineering Group, School of Oral and Dental Sciences, University of Bristol, BS1 2LY (United Kingdom); Kiamehr, Mostafa [Biomaterials and Tissue Engineering Group, Leeds Dental Institute, University of Leeds, LS2 9LU (United Kingdom); Yang, Xuebin [Biomaterials and Tissue Engineering Group, Leeds Dental Institute, University of Leeds, LS2 9LU (United Kingdom); NIHR Leeds Musculoskeletal Biomedical Research Unit, Chapel Allerton Hospital, Leeds LS7 4SA (United Kingdom); Su, Bo, E-mail: b.su@bristol.ac.uk [Biomaterials Engineering Group, School of Oral and Dental Sciences, University of Bristol, BS1 2LY (United Kingdom)

2013-07-01

In this work a bioactive composite scaffold, comprised of bioactive-glass and gelatin, is introduced. Through direct foaming a sponge-like composite of a sol–gel derived bioactive-glass (70S30C; 70% SiO{sub 2}, 30% CaO) and porcine gelatin was developed for use as a biodegradable scaffold for bone tissue engineering. The composite was developed to provide a suitable alternative to synthetic polymer based scaffolds, allowing directed regeneration of bone tissue. The fabricated scaffold was characterised through X-ray microtomography, scanning electron and light microscopy demonstrating a three dimensionally porous and interconnected structure, with an average pore size (170 μm) suitable for successful cell proliferation and tissue ingrowth. Acellular bioactivity was assessed through apatite formation during submersion in simulated body fluid (SBF) whereby the rate and onset of apatite nucleation was found to be comparable to that of bioactive-glass. Modification of dehydrothermal treatment parameters induced varying degrees of crosslinking, allowing the degradation of the composite to be tailored to suit specific applications and establishing its potential for a wide range of applications. Use of genipin to supplement crosslinking by dehydrothermal treatment provided further means of modifying degradability. Biocompatibility of the composite was qualified through successful cultures of human dental pulp stem cells (HDPSCs) on samples of the composite scaffold. Osteogenic differentiation of HDPSCs and extracellular matrix deposition were confirmed through positive alkaline phosphatase staining and immunohistochemistry. - Highlights: ► Optimised composition and fabrication produced sponge-like porosity (pore size ∼ 170 μm). ► Maximum aqueous stability via dehydrothermal treatment at 145 °C, for 48 h ► Biocompatibility and osteogenic potential confirmed via successful HDPSC cultures. ► Minimal toxicity exhibited in optimally crosslinked samples (10 m

Preparation of gelatin based porous biocomposite for bone tissue engineering and evaluation of gamma irradiation effect on its properties

Energy Technology Data Exchange (ETDEWEB)

Islam, Md. Minhajul [Department of Applied Chemistry and Chemical Engineering, Faculty of Engineering and Technology, University of Dhaka, Dhaka 1000 (Bangladesh); Khan, Mubarak A. [Institute of Radiation and Polymer Technology (IRPT), Atomic Energy Research Establishment (AERE), P. O. Box No. 3787, Dhaka 1000 (Bangladesh); Rahman, Mohammed Mizanur, E-mail: mizanur.rahman@du.ac.bd [Department of Applied Chemistry and Chemical Engineering, Faculty of Engineering and Technology, University of Dhaka, Dhaka 1000 (Bangladesh)

2015-04-01

Biodegradable porous hybrid polymer composites were prepared by using gelatin as base polymer matrix, β-tricalcium phosphate (TCP) and calcium sulfate (CS) as cementing materials, chitosan as an antimicrobial agent, and glutaraldehyde and polyethylene glycol (PEG) as crosslinkers at different mass ratios. Thereafter, the composites were subjected to γ-radiation sterilization. The structure and properties of these composite scaffolds were characterized by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), mechanical properties testing (compressive, bending, tensile and impact), thermogravimetry/differential thermal analysis (TG/DTA), and physical stability test in simulated body fluid (SBF). We found that TCP rich composites showed enhanced mechanical properties among all the crosslinked composites. γ-Radiation sterilization triggered further cross linking in polymer matrix resulting a decrease in pore size of the composites and an increase in pore wall thickness with improved mechanical and thermal properties. The chemically crosslinked composite with 40% TCP followed by γ-radiation sterilization showed the smallest pore size distribution with a mean pore diameter of 159.22 μm, which falls in the range of 100–350 μm — known to be suitable for osteoconduction. Considering its improved mechanical and thermal properties along with osteoconduction ability without cytotoxicity, we propose this biocomposite as a viable candidate for bone tissue engineering. - Highlights: • Composite scaffolds were prepared from biopolymers (gelatin and chitosan). • β-TCP and CS were used as bioactive cementing materials at different ratios. • γ-Sterilization improved the mechanical properties of the biocomposites. • γ-Sterilization reduced the cytotoxicity and induced high antimicrobial properties. • Composite having 40% TCP has the proper pore size distribution for osteoconduction.

Computational segmentation of collagen fibers in bone matrix indicates bone quality in ovariectomized rat spine.

Science.gov (United States)

Daghma, Diaa Eldin S; Malhan, Deeksha; Simon, Paul; Stötzel, Sabine; Kern, Stefanie; Hassan, Fathi; Lips, Katrin Susanne; Heiss, Christian; El Khassawna, Thaqif

2018-05-01

Bone loss varies according to disease and age and these variations affect bone cells and extracellular matrix. Osteoporosis rat models are widely investigated to assess mechanical and structural properties of bone; however, bone matrix proteins and their discrepant regulation of diseased and aged bone are often overlooked. The current study considered the spine matrix properties of ovariectomized rats (OVX) against control rats (Sham) at 16 months of age. Diseased bone showed less compact structure with inhomogeneous distribution of type 1 collagen (Col1) and changes in osteocyte morphology. Intriguingly, demineralization patches were noticed in the vicinity of blood vessels in the OVX spine. The organic matrix structure was investigated using computational segmentation of collagen fibril properties. In contrast to the aged bone, diseased bone showed longer fibrils and smaller orientation angles. The study shows the potential of quantifying transmission electron microscopy images to predict the mechanical properties of bone tissue.

Characterization of an Olive Flounder Bone Gelatin-Zinc Oxide Nanocomposite Film and Evaluation of Its Potential Application in Spinach Packaging.

Science.gov (United States)

Beak, Songee; Kim, Hyeri; Song, Kyung Bin

2017-11-01

Olive flounder bone gelatin (OBG) was used for a film base material in this study. In addition, zinc oxide nanoparticles (ZnO) were incorporated into the OBG film to prepare a nanocomposite film and to impart antimicrobial activity to it. The tensile strength of the OBG film increased by 6.62 MPa, and water vapor permeability and water solubility decreased by 0.93 × 10 -9 g/m s Pa and 13.79%, respectively, by the addition of ZnO to the OBG film. In particular, the OBG-ZnO film exhibited antimicrobial activity against Listeria monocytogenes. To investigate the applicability of the OBG-ZnO packaging film, fresh spinach was wrapped in this film and stored for a week. The results indicated that the OBG-ZnO film showed antimicrobial activity against L. monocytogenes inoculated on spinach without affecting the quality of spinach, such as vitamin C content and color. Thus, the OBG-ZnO nanocomposite film can be applied as an efficient antimicrobial food packaging material. As a base material of edible films, gelatin was extracted from olive flounder bone, which is fish processing by-product. Olive flounder bone gelatin (OBG) nanocomposite films were prepared with zinc oxide nanoparticles (ZnO). For an application to antimicrobial packaging, spinach was wrapped with the OBG-ZnO nanocomposite film. © 2017 Institute of Food Technologists®.

Comparison of the behavior of fibroblast and bone marrow-derived mesenchymal stem cell on nitrogen plasma-treated gelatin films

International Nuclear Information System (INIS)

Prasertsung, I.; Kanokpanont, S.; Mongkolnavin, R.; Wong, C.S.; Panpranot, J.; Damrongsakkul, S.

2013-01-01

The attachment and growth behavior of mouse fibroblast (L929) and rat bone marrow-derived mesenchymal stem cell (MSC) on nitrogen plasma-treated and untreated gelatin films was investigated and compared. The gelatin films were prepared by solution casting (0.05% w/v) and crosslinked using dehydrothermal treatment. The crosslinked gelatin films were treated with nitrogen alternating current (AC) 50 Hz plasma systems at various treatment time. The results on the attachment and growth of two cells; L929 and MSC, on plasma-treated gelatin film showed that the number of attached and proliferated cells on plasma-treated gelatin films was significantly increased compared to untreated samples. However, no significant difference between the number of attached L929 and MSC on plasma-treated gelatin was observed. The shorter population doubling time and higher growth rate of cells cultured on plasma-treated film indicated the greater growth of cells, compared to ones on untreated films. The greatest enhancement of cell attachment and growth were noticed when the film was treated with nitrogen plasma for 9 to 15 s. This suggested that the greater attachment and growth of both cells on gelatin films resulted from the change of surface properties, i.e. hydrophilicity, surface energy, and chemistry. The suitable water contact angle and oxygen/nitrogen ratio (O/N) of gelatin film for best L929 and MSC attachment were observed at 27–32° and 1.4, respectively. These conditions also provided the best proliferation of cells on plasma-treated gelatin films. - Highlights: • We compared the attachment and growth behavior of L929 and MSC. • The attachment of two cells on plasma-treated gelatin was significantly increased. • The shorter population doubling time and higher growth rate of cells were observed. • L929 fibroblast exhibited the greater proliferation, compared to MSC

3D chitosan-gelatin-chondroitin porous scaffold improves osteogenic differentiation of mesenchymal stem cells

Energy Technology Data Exchange (ETDEWEB)

Machado, C B [Department of Biochemistry and Immunology, Institute of Biological Sciences, Federal University of Minas Gerais (Brazil); Ventura, J M G [Department of Ceramics and Glass Engineering, University of Aveiro (Portugal); Lemos, A F [Department of Ceramics and Glass Engineering, University of Aveiro (Portugal); Ferreira, J M F [Department of Ceramics and Glass Engineering, University of Aveiro (Portugal); Leite, M F [Department of Physiology and Biophysics, Institute of Biological Sciences, Federal University of Minas Gerais (Brazil); Goes, A M [Department of Biochemistry and Immunology, Institute of Biological Sciences, Federal University of Minas Gerais (Brazil)

2007-06-01

A porous 3D scaffold was developed to support and enhance the differentiation process of mesenchymal stem cells (MSC) into osteoblasts in vitro. The 3D scaffold was made with chitosan, gelatin and chondroitin and it was crosslinked by EDAC. The scaffold physicochemical properties were evaluated. SEM revealed the high porosity and interconnection of pores in the scaffold; rheological measurements show that the scaffold exhibits a characteristic behavior of strong gels. The elastic modulus found in compressive tests of the crosslinked scaffold was about 50 times higher than the non-crosslinked one. After 21 days, the 3D matrix submitted to hydrolytic degradation loses above 40% of its weight. MSC were collected from rat bone marrow and seeded in chitosan-gelatin-chondroitin 3D scaffolds and in 2D culture plates as well. MSC were differentiated into osteoblasts for 21 days. Cell proliferation and alkaline phosphatase activity were followed weekly during the osteogenic process. The osteogenic differentiation of MSC was improved in 3D culture as shown by MTT assay and alkaline phosphatase activity. On the 21st day, bone markers, osteopontin and osteocalcin, were detected by the PCR analysis. This study shows that the chitosan-gelatin-chondroitin 3D structure provides a good environment for the osteogenic process and enhances cellular proliferation.

3D chitosan-gelatin-chondroitin porous scaffold improves osteogenic differentiation of mesenchymal stem cells.

Science.gov (United States)

Machado, C B; Ventura, J M G; Lemos, A F; Ferreira, J M F; Leite, M F; Goes, A M

2007-06-01

A porous 3D scaffold was developed to support and enhance the differentiation process of mesenchymal stem cells (MSC) into osteoblasts in vitro. The 3D scaffold was made with chitosan, gelatin and chondroitin and it was crosslinked by EDAC. The scaffold physicochemical properties were evaluated. SEM revealed the high porosity and interconnection of pores in the scaffold; rheological measurements show that the scaffold exhibits a characteristic behavior of strong gels. The elastic modulus found in compressive tests of the crosslinked scaffold was about 50 times higher than the non-crosslinked one. After 21 days, the 3D matrix submitted to hydrolytic degradation loses above 40% of its weight. MSC were collected from rat bone marrow and seeded in chitosan-gelatin-chondroitin 3D scaffolds and in 2D culture plates as well. MSC were differentiated into osteoblasts for 21 days. Cell proliferation and alkaline phosphatase activity were followed weekly during the osteogenic process. The osteogenic differentiation of MSC was improved in 3D culture as shown by MTT assay and alkaline phosphatase activity. On the 21st day, bone markers, osteopontin and osteocalcin, were detected by the PCR analysis. This study shows that the chitosan-gelatin-chondroitin 3D structure provides a good environment for the osteogenic process and enhances cellular proliferation.

3D chitosan-gelatin-chondroitin porous scaffold improves osteogenic differentiation of mesenchymal stem cells

International Nuclear Information System (INIS)

Machado, C B; Ventura, J M G; Lemos, A F; Ferreira, J M F; Leite, M F; Goes, A M

2007-01-01

A porous 3D scaffold was developed to support and enhance the differentiation process of mesenchymal stem cells (MSC) into osteoblasts in vitro. The 3D scaffold was made with chitosan, gelatin and chondroitin and it was crosslinked by EDAC. The scaffold physicochemical properties were evaluated. SEM revealed the high porosity and interconnection of pores in the scaffold; rheological measurements show that the scaffold exhibits a characteristic behavior of strong gels. The elastic modulus found in compressive tests of the crosslinked scaffold was about 50 times higher than the non-crosslinked one. After 21 days, the 3D matrix submitted to hydrolytic degradation loses above 40% of its weight. MSC were collected from rat bone marrow and seeded in chitosan-gelatin-chondroitin 3D scaffolds and in 2D culture plates as well. MSC were differentiated into osteoblasts for 21 days. Cell proliferation and alkaline phosphatase activity were followed weekly during the osteogenic process. The osteogenic differentiation of MSC was improved in 3D culture as shown by MTT assay and alkaline phosphatase activity. On the 21st day, bone markers, osteopontin and osteocalcin, were detected by the PCR analysis. This study shows that the chitosan-gelatin-chondroitin 3D structure provides a good environment for the osteogenic process and enhances cellular proliferation

Conductive vancomycin-loaded mesoporous silica polypyrrole-based scaffolds for bone regeneration

DEFF Research Database (Denmark)

Zanjanizadeh Ezazi, Nazanin; Shahbazi, Mohammad-Ali; Shatalin, Yuri V.

2018-01-01

-conductive scaffolds. Osteoblast cells were perfectly immersed into the gelatin matrix and remained viable for 14 days. Overall, new conductive composite bone scaffolds were created and the obtained results strongly verified the applicability of this conductive scaffold in drug delivery, encouraging its further...

Influence of short-term aluminum exposure on demineralized bone matrix induced bone formation

Energy Technology Data Exchange (ETDEWEB)

Severson, A.R. (Minnesota Univ., Duluth, MN (United States). Dept. of Anatomy and Cell Biology); Haut, C.F.; Firling, C.E. (Minnesota Univ., Duluth, MN (United States). Dept. of Biology); Huntley, T.E. (Minnesota Univ., Duluth, MN (United States). Dept. of Biochemistry and Molecular Biology)

1992-12-01

The effects of aluminum exposure on bone formation employing the demineralized bone matrix (DBM) induced bone development model were studied using 4-week-old Sprague-Dawley rats injected with a saline (control) or an aluminum chloride (experimental) solution. After 2 weeks of aluminum treatment, 20-mg portions of rat DBM were implanted subcutaneously on each side in the thoracic region of the control and experimental rats. Animals were killed 7, 12, or 21 days after implantation of the DBM and the developing plaques removed. No morphological, histochemical, or biochemical differences were apparent between plaques from day 7 control and experimental rats. Plaques from day 12 control and experimental rats exhibited cartilage formation and alkaline phosphatase activity localized in osteochondrogenic cells, chondrocytes, osteoblasts, and extracellular matrix. Unlike the plaques from control rats that contained many osteoblastic mineralizing fronts, the plaques from the 12-day experimental group had a preponderance of cartilaginous tissue, no evidence of mineralization, increased levels of alkaline phosphatase activity, and a reduced calcium content. Plaques developing for 21 days in control animals demonstrated extensive new bone formation and bone marrow development, while those in the experimental rats demonstrated unmineralized osteoid-like matrix with poorly developed bone marrow. Alkaline phosphatase activity of the plaques continued to remain high on day 21 for the control and experimental groups. Calcium levels were significantly reduced in the experimental group. These biochemical changes correlated with histochemical reductions in bone calcification. Thus, aluminum administration to rats appears to alter the differentiation and calcification of developing cartilage and bone in the DBM-induced bone formation model and suggests that aluminum by some mechanism alters the matrix calcification in growing bones. (orig.).

Osteoblast Differentiation and Bone Matrix Formation In Vivo and In Vitro.

Science.gov (United States)

Blair, Harry C; Larrouture, Quitterie C; Li, Yanan; Lin, Hang; Beer-Stoltz, Donna; Liu, Li; Tuan, Rocky S; Robinson, Lisa J; Schlesinger, Paul H; Nelson, Deborah J

2017-06-01

We review the characteristics of osteoblast differentiation and bone matrix synthesis. Bone in air breathing vertebrates is a specialized tissue that developmentally replaces simpler solid tissues, usually cartilage. Bone is a living organ bounded by a layer of osteoblasts that, because of transport and compartmentalization requirements, produce bone matrix exclusively as an organized tight epithelium. With matrix growth, osteoblasts are reorganized and incorporated into the matrix as living cells, osteocytes, which communicate with each other and surface epithelium by cell processes within canaliculi in the matrix. The osteoblasts secrete the organic matrix, which are dense collagen layers that alternate parallel and orthogonal to the axis of stress loading. Into this matrix is deposited extremely dense hydroxyapatite-based mineral driven by both active and passive transport and pH control. As the matrix matures, hydroxyapatite microcrystals are organized into a sophisticated composite in the collagen layer by nucleation in the protein lattice. Recent studies on differentiating osteoblast precursors revealed a sophisticated proton export network driving mineralization, a gene expression program organized with the compartmentalization of the osteoblast epithelium that produces the mature bone matrix composite, despite varying serum calcium and phosphate. Key issues not well defined include how new osteoblasts are incorporated in the epithelial layer, replacing those incorporated in the accumulating matrix. Development of bone in vitro is the subject of numerous projects using various matrices and mesenchymal stem cell-derived preparations in bioreactors. These preparations reflect the structure of bone to variable extents, and include cells at many different stages of differentiation. Major challenges are production of bone matrix approaching the in vivo density and support for trabecular bone formation. In vitro differentiation is limited by the organization and

Biomimetic composite coating on rapid prototyped scaffolds for bone tissue engineering.

Science.gov (United States)

Arafat, M Tarik; Lam, Christopher X F; Ekaputra, Andrew K; Wong, Siew Yee; Li, Xu; Gibson, Ian

2011-02-01

The objective of this present study was to improve the functional performance of rapid prototyped scaffolds for bone tissue engineering through biomimetic composite coating. Rapid prototyped poly(ε-caprolactone)/tri-calcium phosphate (PCL/TCP) scaffolds were fabricated using the screw extrusion system (SES). The fabricated PCL/TCP scaffolds were coated with a carbonated hydroxyapatite (CHA)-gelatin composite via biomimetic co-precipitation. The structure of the prepared CHA-gelatin composite coating was studied by scanning electron microscopy (SEM), X-ray photoelectron spectroscopy and Fourier transform infrared spectroscopy. Compressive mechanical testing revealed that the coating process did not have any detrimental effect on the mechanical properties of the scaffolds. The cell-scaffold interaction was studied by culturing porcine bone marrow stromal cells (BMSCs) on the scaffolds and assessing the proliferation and bone-related gene and protein expression capabilities of the cells. Confocal laser microscopy and SEM images of the cell-scaffold constructs showed a uniformly distributed cell sheet and accumulation of extracellular matrix in the interior of CHA-gelatin composite-coated PCL/TCP scaffolds. The proliferation rate of BMSCs on CHA-gelatin composite-coated PCL/TCP scaffolds was about 2.3 and 1.7 times higher than that on PCL/TCP scaffolds and CHA-coated PCL/TCP scaffolds, respectively, by day 10. Furthermore, reverse transcription polymerase chain reaction and Western blot analysis revealed that CHA-gelatin composite-coated PCL/TCP scaffolds stimulate osteogenic differentiation of BMSCs the most, compared with PCL/TCP scaffolds and CHA-coated PCL/TCP scaffolds. These results demonstrate that CHA-gelatin composite-coated rapid prototyped PCL/TCP scaffolds are promising for bone tissue engineering. Copyright © 2010 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Minocycline-released hydroxyapatite-gelatin nanocomposite and its cytocompatibility in vitro

Energy Technology Data Exchange (ETDEWEB)

Dou Xiaochen; Zhu Xiaopeng; Zhou Jian; Cai Huaqiong; Li Quanli [Key Laboratory of Oral Disease Research of Anhui Province, Stomatologic Hospital and Collage, Anhui Medical University, Hefei (China); Tang Jian, E-mail: ql-li@126.com [Department of Orthopedic Surgery, The First Hospital of Anhui Medical University, Hefei (China)

2011-04-15

The incorporation of antibacterial agents into biomaterials is extremely desirable for repairing bone defects. Minocycline, a semi-synthetic tetracycline antibiotic, is active against aerobic, anaerobic, Gram-positive and Gram-negative bacteria, and can enhance bone formation, decrease connective tissue breakdown and diminish bone resorption. In this study, a novel minocycline-releasing biomaterial was synthesized using a biomimetic method. A measured amount of an acidic hydroxyapatite and minocycline solution was respectively added to a gelatin solution and kept at 40 deg. C and pH 7-8 for 2 h. The mixture was aged overnight, lyophilized and a hydroxyapatite-gelatin-minocycline composite was obtained. The composite was co-cultured with rat bone marrow stromal cells (BMSCs) in vitro. Our results show that nanohydroxyapatite was distributed evenly in the fibrils of the gelatin. Minocycline was incorporated into the composite and could be released from the composite particles slowly over 2 weeks in vitro. The composite promoted BMSC adhesion, proliferation and differentiation in vitro. The approach described here may provide a basis for the preparation of an antibacterial biomaterial for bone regeneration.

Minocycline-released hydroxyapatite-gelatin nanocomposite and its cytocompatibility in vitro

International Nuclear Information System (INIS)

Dou Xiaochen; Zhu Xiaopeng; Zhou Jian; Cai Huaqiong; Li Quanli; Tang Jian

2011-01-01

The incorporation of antibacterial agents into biomaterials is extremely desirable for repairing bone defects. Minocycline, a semi-synthetic tetracycline antibiotic, is active against aerobic, anaerobic, Gram-positive and Gram-negative bacteria, and can enhance bone formation, decrease connective tissue breakdown and diminish bone resorption. In this study, a novel minocycline-releasing biomaterial was synthesized using a biomimetic method. A measured amount of an acidic hydroxyapatite and minocycline solution was respectively added to a gelatin solution and kept at 40 deg. C and pH 7-8 for 2 h. The mixture was aged overnight, lyophilized and a hydroxyapatite-gelatin-minocycline composite was obtained. The composite was co-cultured with rat bone marrow stromal cells (BMSCs) in vitro. Our results show that nanohydroxyapatite was distributed evenly in the fibrils of the gelatin. Minocycline was incorporated into the composite and could be released from the composite particles slowly over 2 weeks in vitro. The composite promoted BMSC adhesion, proliferation and differentiation in vitro. The approach described here may provide a basis for the preparation of an antibacterial biomaterial for bone regeneration.

Enhanced Bone Tissue Regeneration by Porous Gelatin Composites Loaded with the Chinese Herbal Decoction Danggui Buxue Tang.

Directory of Open Access Journals (Sweden)

Wen-Ling Wang

Full Text Available Danggui Buxue Tang (DBT is a traditional Chinese herbal decoction containing Radix Astragali and Radix Angelicae sinensis. Pharmacological results indicate that DBT can stimulate bone cell proliferation and differentiation. The aim of the study was to investigate the efficacy of adding DBT to bone substitutes on bone regeneration following bone injury. DBT was incorporated into porous composites (GGT made from genipin-crosslinked gelatin and β-triclacium phosphates as bone substitutes (GGTDBT. The biological response of mouse calvarial bone to these composites was evaluated by in vivo imaging systems (IVIS, micro-computed tomography (micro-CT, and histology analysis. IVIS images revealed a stronger fluorescent signal in GGTDBT-treated defect than in GGT-treated defect at 8 weeks after implantation. Micro-CT analysis demonstrated that the level of repair from week 4 to 8 increased from 42.1% to 71.2% at the sites treated with GGTDBT, while that increased from 33.2% to 54.1% at GGT-treated sites. These findings suggest that the GGTDBT stimulates the innate regenerative capacity of bone, supporting their use in bone tissue regeneration.

Coherent scattering and matrix correction in bone-lead measurements

International Nuclear Information System (INIS)

Todd, A.C.

2000-01-01

The technique of K-shell x-ray fluorescence of lead in bone has been used in many studies of the health effects of lead. This paper addresses one aspect of the technique, namely the coherent conversion factor (CCF) which converts between the matrix of the calibration standards and those of human bone. The CCF is conventionally considered a constant but is a function of scattering angle, energy and the elemental composition of the matrices. The aims of this study were to quantify the effect on the CCF of several assumptions which may not have been tested adequately and to compare the CCFs for plaster of Paris (the present matrix of calibration standards) and a synthetic apatite matrix. The CCF was calculated, using relativistic form factors, for published compositions of bone, both assumed and assessed compositions of plaster, and the synthetic apatite. The main findings of the study were, first, that impurities in plaster, lead in the plaster or bone matrices, coherent scatter from non-bone tissues and the individual subject's measurement geometry are all minor or negligible effects; and, second, that the synthetic apatite matrix is more representative of bone mineral than is plaster of Paris. (author)

Promotion of osteogenic differentiation of stem cells and increase of bone-bonding ability in vivo using urease-treated titanium coated with calcium phosphate and gelatin

Science.gov (United States)

Huang, Zhong-Ming; Qi, Yi-Ying; Du, Shao-Hua; Feng, Gang; Unuma, Hidero; Yan, Wei-Qi

2013-10-01

Because of its excellent biocompatibility and low allergenicity, titanium has been widely used for bone replacement and tissue engineering. To produce a desirable composite with enhanced bone response and mechanical strength, in this study bioactive calcium phosphate (CaP) and gelatin composites were coated onto titanium (Ti) via a novel urease technique. The cellular responses to the CaP/gelatin/Ti (CaP/gel/Ti) and bone bonding ability were evaluated with proliferation and osteogenic differentiation of mesenchymal stem cells (MSCs) on CaP/gel/Ti and CaP/Ti in vitro. The results showed that the optical density values, alkaline phosphatase expression and genes expression of MSCs on CaP/gel/Ti were similar to those on CaP/Ti, yet significantly higher than those on pure Ti (p layer. An interfacial layer, containing Ti, Ca and P, was found to form at the interface between bone and the implant on all three groups by EDS analysis. However, the content of Ca, P in the surface of CaP/gel/Ti implants was more than in the other two groups at each time point. The CaP/gel/Ti modified by the urease method was not only beneficial for MSCs proliferation and osteogenic differentiation, but also favorable for bone bonding ability on Ti implants in vivo, suggesting that Ti functionalized with CaP and gelatin might have a great potential in clinical joint replacement or dental implants.

Osteogenic Matrix Cell Sheets Facilitate Osteogenesis in Irradiated Rat Bone

Directory of Open Access Journals (Sweden)

Yoshinobu Uchihara

2015-01-01

Full Text Available Reconstruction of large bone defects after resection of malignant musculoskeletal tumors is a significant challenge in orthopedic surgery. Extracorporeal autogenous irradiated bone grafting is a treatment option for bone reconstruction. However, nonunion often occurs because the osteogenic capacity is lost by irradiation. In the present study, we established an autogenous irradiated bone graft model in the rat femur to assess whether osteogenic matrix cell sheets improve osteogenesis of the irradiated bone. Osteogenic matrix cell sheets were prepared from bone marrow-derived stromal cells and co-transplanted with irradiated bone. X-ray images at 4 weeks after transplantation showed bridging callus formation around the irradiated bone. Micro-computed tomography images at 12 weeks postoperatively showed abundant callus formation in the whole circumference of the irradiated bone. Histology showed bone union between the irradiated bone and host femur. Mechanical testing showed that the failure force at the irradiated bone site was significantly higher than in the control group. Our study indicates that osteogenic matrix cell sheet transplantation might be a powerful method to facilitate osteogenesis in irradiated bones, which may become a treatment option for reconstruction of bone defects after resection of malignant musculoskeletal tumors.

Treatment of active unicameral bone cysts with percutaneous injection of demineralized bone matrix and autogenous bone marrow.

Science.gov (United States)

Rougraff, Bruce T; Kling, Thomas J

2002-06-01

The treatment of unicameral bone cysts varies from open bone-grafting procedures to percutaneous injection of corticosteroids or bone marrow. The purpose of this study was to evaluate the feasibility and effectiveness of percutaneous injection of a mixture of demineralized bone matrix and autogenous bone marrow for the treatment of simple bone cysts. Twenty-three patients with an active unicameral bone cyst were treated with trephination and injection of allogeneic demineralized bone matrix and autogenous bone marrow. The patients were followed for an average of fifty months (range, thirty to eighty-one months), at which time pain, function, and radiographic signs of resolution of the cyst were assessed. The average time until the patients had pain relief was five weeks, and the average time until the patients returned to full, unrestricted activities was six weeks. Bone-healing at the site of the injection was first seen radiographically at three to six months. No patient had a pathologic fracture during this early bone-healing stage. Cortical remodeling was seen radiographically by six to nine months, and after one year the response was usually complete, changing very little from then on. Five patients required a second injection because of recurrence of the cyst, and all five had a clinically and radiographically quiescent cyst after an average of thirty-six additional months of follow-up. Seven of the twenty-three patients had incomplete healing manifested by small, persistent radiolucent areas within the original cyst. None of these cysts increased in size or resulted in pain or fracture. Percutaneous injection of allogeneic demineralized bone matrix and autogenous bone marrow is an effective treatment for unicameral bone cysts.

A H2O2 Biosensor Based on Immobilization of HorseradishPeroxidase in a Gelatine Network Matrix

Directory of Open Access Journals (Sweden)

Jun-Jie Zhu

2005-05-01

Full Text Available A simple and promising H2O2 biosensor has been developed by successfulentrapment of horseradish peroxidase (HRP in a gelatine matrix which was cross-linkedwith formaldehyde. The large microscopic surface area and porous morphology of thegelatine matrix lead to high enzyme loading and the enzyme entrapped in this matrix canretain its bioactivity. This biosensor exhibited a fast amperometric response to hydrogenperoxide (H2O2. The linear range for H2O2 determination was from 2.5×10-5 to2.5×10-3 M, with a detection limit of 2.0×10-6 M based on S / N = 3. This biosensorpossessed very good reproducibility.

Photoluminescence and structural properties of CdSe quantum dot–gelatin composite films

Energy Technology Data Exchange (ETDEWEB)

Borkovska, L., E-mail: bork@isp.kiev.ua [V. Lashkaryov Institute of Semiconductor Physics of NAS of Ukraine, Pr. Nauky 41, 03028 Kyiv (Ukraine); Korsunska, N.; Stara, T.; Gudymenko, O.; Kladko, V. [V. Lashkaryov Institute of Semiconductor Physics of NAS of Ukraine, Pr. Nauky 41, 03028 Kyiv (Ukraine); Stroyuk, O.; Raevskaya, A. [L. Pysarzhevsky Institute of Physical Chemistry of NAS of Ukraine, Pr. Nauky 31, 03028 Kyiv (Ukraine); Kryshtab, T. [Instituto Politécnico Nacional – ESFM, Av. IPN, Ed.9 U.P.A.L.M., 07738 Mexico D.F. (Mexico)

2014-11-15

Optical and structural properties of composite films of CdSe quantum dots (QDs) embedded in gelatin matrix have been investigated by photoluminescence (PL), optical absorption and X-ray diffraction (XRD) methods. The optical absorption of the composite in the visible spectral range is found to be determined mainly by light absorption in the QDs. The decrease of the film transparency and the shift of the absorption edge to lower energies observed upon thermal annealing of the films at 140–160 °C are ascribed to the formation of chromophore groups in gelatin matrix. XRD patterns of the composite revealed helix to coil transition in gelatin matrix under thermal annealing of the composite at 100–160 °C. It is found that PL spectra of the composite are dominated by exciton and defect-related emission of the QDs and also contain weak emission of gelatin matrix. It is found that thermal annealing of the composite at 100–160 °C changes PL intensity and produces the shift of the PL bands to lower energies. As the annealed composite was kept in air for several months, the shift of exciton-related PL band position restored partially and the PL intensity increased. It is proposed that the increase of the PL intensity upon the thermal annealing of composite at 140 °C can be used for enhancement of the QD-related PL. Changes that occurred in the PL spectra of composite are ascribed to structural and chemical transformations in gelatin matrix and at the QD/gelatin interface.

Enhanced release of bone morphogenetic proteins from demineralized bone matrix by gamma irradiation

International Nuclear Information System (INIS)

Sung, Nak-Yun; Choi, Jong-il

2015-01-01

Gamma irradiation is a useful method for sterilizing demineralized bone matrix (DBM), but its effect on the osteoinductivity of DBM is still controversial. In this study, the osteoinductive activity of gamma-irradiated DBM was examined using a mouse myoblastic cell line (C2C12). DBM was extracted from adult bovine bone and was irradiated at a dose of 25 kGy using a 60 cobalt gamma-irradiator. Cell proliferation with DBM was not affected by gamma-irradiation, but alkaline phosphatase and osteocalcin productions were significantly increased in C2C12 cell groups treated with gamma-irradiated DBM. It was reasoned that bone morphogenetic proteins were more efficiently released from gamma-irradiated DBM than from the non-irradiated control. This result suggests the effectiveness of radiation sterilization of bone implants - Highlights: • Demineralized bone matrix (DBM) was gamma-irradiated for sterilization. • Irradiated DBM had higher alkaline phosphatase and osteocalcin production. • It was reasoned the more released bone morphogenetic proteins by irradiation. • This result supports the application of radiation sterilization for bone implants

The physico-chemical properties of pangas catfish (Pangasius pangasius) skin gelatin

Science.gov (United States)

Pradarameswari, K. A.; Zaelani, K.; Waluyo, E.; Nurdiani, R.

2018-04-01

Gelatin can be used as emulsifier and stabilizer in food products. Until now, the most widely used raw materials for the production of gelatin industry are cow bone, cow skin and pig skin. Fish gelatin has been highlighted as a better alternative to replace mammals gelatin based on ethical and religious perspective. Fish gelatin was extracted from Pangas catfish skin to determine its physico-chemical properties. Different temperatures (45 °C, 50 °C, 55 °C) were employed during gelatin extraction. Higher temperature increased the yield and fat contents of Pangas catfish skin gelatin. In contrary, higher water, protein, ash contents were observed during lower temperature. Temperature significantly (p fish skin gelatin. Based on the FTIR spectrum catfish skin gelatin functional groups can be identified as N-H, O-H, C = H, C-O and C-H.

Preparation of Rhodamine B Fluorescent Poly(methacrylic acid) Coated Gelatin Nanoparticles

OpenAIRE

Gan, Zhenhai; Ju, Jianhui; Zhang, Ting; Wu, Daocheng

2011-01-01

Poly(methacrylic acid) (PMAA)-coated gelatin nanoparticles encapsulated with fluorescent dye rhodamine B were prepared by the coacervation method with the aim to retard the release of rhodamine B from the gelatin matrix. With sodium sulfate as coacervation reagent for gelatin, a kind of biopolymer with excellent biocompatibility, the formed gelatin nanoparticles were cross-linked by formaldehyde followed by the polymerization of methacrylic acid coating. The fluorescent poly(methacrylic acid)...

In vitro and in vivo Biocompatibility of Alginate Dialdehyde/Gelatin Hydrogels with and without Nanoscaled Bioactive Glass for Bone Tissue Engineering Applications

Directory of Open Access Journals (Sweden)

Ulrike Rottensteiner

2014-03-01

Full Text Available In addition to good mechanical properties needed for three-dimensional tissue engineering, the combination of alginate dialdehyde, gelatin and nano-scaled bioactive glass (45S5 is supposed to combine excellent cellular adhesion, proliferation and differentiation properties, good biocompatibility and predictable degradation rates. The goal of this study was to evaluate the in vitro and in vivo biocompatibility as a first step on the way to its use as a scaffold in bone tissue engineering. In vitro evaluation showed good cell adherence and proliferation of bone marrow derived mesenchymal stem cells seeded on covalently crosslinked alginate dialdehyde-gelatin (ADA-GEL hydrogel films with and without 0.1% nano-Bioglass® (nBG. Lactate dehydrogenase (LDH- and mitochondrial activity significantly increased in both ADA-GEL and ADA-GEL-nBG groups compared to alginate. However, addition of 0.1% nBG seemed to have slight cytotoxic effect compared to ADA-GEL. In vivo implantation did not produce a significant inflammatory reaction, and ongoing degradation could be seen after four weeks. Ongoing vascularization was detected after four weeks. The good biocompatibility encourages future studies using ADA-GEL and nBG for bone tissue engineering application.

Promotion of osteogenic differentiation of stem cells and increase of bone-bonding ability in vivo using urease-treated titanium coated with calcium phosphate and gelatin

International Nuclear Information System (INIS)

Huang, Zhong-Ming; Qi, Yi-Ying; Du, Shao-Hua; Feng, Gang; Yan, Wei-Qi; Unuma, Hidero

2013-01-01

Because of its excellent biocompatibility and low allergenicity, titanium has been widely used for bone replacement and tissue engineering. To produce a desirable composite with enhanced bone response and mechanical strength, in this study bioactive calcium phosphate (CaP) and gelatin composites were coated onto titanium (Ti) via a novel urease technique. The cellular responses to the CaP/gelatin/Ti (CaP/gel/Ti) and bone bonding ability were evaluated with proliferation and osteogenic differentiation of mesenchymal stem cells (MSCs) on CaP/gel/Ti and CaP/Ti in vitro. The results showed that the optical density values, alkaline phosphatase expression and genes expression of MSCs on CaP/gel/Ti were similar to those on CaP/Ti, yet significantly higher than those on pure Ti (p < 0.05). CaP/gel/Ti and CaP/Ti rods (2 mm in diameter, 10 mm in length) were also implanted into femoral shaft of rabbits and pure Ti rods served as control (n = 10). Histological examination, scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS) measurements were performed at 4 and 8 weeks after the operation. The histological and SEM observations demonstrated clearly that more new bone formed on the surface of CaP/gel/Ti than in the other two groups at each time point. The CaP/gel/Ti bonded to the surrounding bone directly with no intervening soft tissue layer. An interfacial layer, containing Ti, Ca and P, was found to form at the interface between bone and the implant on all three groups by EDS analysis. However, the content of Ca, P in the surface of CaP/gel/Ti implants was more than in the other two groups at each time point. The CaP/gel/Ti modified by the urease method was not only beneficial for MSCs proliferation and osteogenic differentiation, but also favorable for bone bonding ability on Ti implants in vivo, suggesting that Ti functionalized with CaP and gelatin might have a great potential in clinical joint replacement or dental implants. (paper)

Efficacy of supermacroporous poly(ethylene glycol)–gelatin cryogel matrix for soft tissue engineering applications

Energy Technology Data Exchange (ETDEWEB)

Sharma, Archana [Department of Biological Sciences, Birla Institute of Technology and Science, Pilani-K.K Birla Goa Campus, 403726 Goa (India); Bhat, Sumrita [Department of Biological Sciences and Bioengineering, Indian Institute of Technology Kanpur, Kanpur, 208016 UP (India); Nayak, Vijayashree, E-mail: vijayashree@goa.bits-pilani.ac.in [Department of Biological Sciences, Birla Institute of Technology and Science, Pilani-K.K Birla Goa Campus, 403726 Goa (India); Kumar, Ashok, E-mail: ashokkum@iitk.ac.in [Department of Biological Sciences and Bioengineering, Indian Institute of Technology Kanpur, Kanpur, 208016 UP (India)

2015-02-01

Three dimensional scaffolds synthesized using natural or synthetic polymers act as an artificial niche for cell adherence and proliferation. In this study, we have fabricated cryogels employing blend of poly (ethylene glycol) (PEG) and gelatin using two different crosslinkers like, glutaraldehyde and EDC-NHS by cryogelation technique. Synthesized matrices possessed interconnected porous structure in the range of 60–100 μm diameter and regained their original length after 90% compression without deformation. Visco-elastic behavior was studied by rheology and unconfined compression analysis, elastic modulus of these cryogels was observed to be > 10{sup 5} Pa which showed their elasticity and mechanical strength. TGA and DSC also showed the stability of these cryogels at different temperatures. In vitro degradation capacity was analyzed for 4 weeks at 37 °C. IMR-32, C2C12 and Cos-7 cells proliferation and ECM secretion on PEG–gelatin cryogels were observed by SEM and fluorescent analysis. In vitro biocompatibility was analyzed by MTT assay for the period of 15 days. Furthermore, cell proliferation efficiency, metabolic activity and functionality of IMR-32 cells were analyzed by neurotransmitter assay and DNA quantification. The cell–matrix interaction, elasticity, mechanical strength, stability at different temperatures, biocompatible, degradable nature showed the potentiality of these cryogels towards soft tissue engineering such as neural, cardiac and skin. - Highlights: • PEG–gelatin cryogel matrices were produced by cryogelation technology. • Matrices showed suitable properties for tissue engineering applications. • Polymeric cryogels supported growth of IMR-32, C2C12 and Cos-7 cells in vitro.

Radiocarbon dating of VIRI bone samples using ultrafiltration

Energy Technology Data Exchange (ETDEWEB)

Minami, Masayo, E-mail: minami@nendai.nagoya-u.ac.jp [Center for Chronological Research, Nagoya University, Nagoya 464-8602 (Japan); Yamazaki, Kana [Faculty of Science, Nagoya University, Nagoya 464-8602 (Japan); Omori, Takayuki [University Museum, University of Tokyo, Tokyo 113-0033 (Japan); Nakamura, Toshio [Center for Chronological Research, Nagoya University, Nagoya 464-8602 (Japan)

2013-01-15

Ultrafiltration can effectively remove low-molecular-weight (LMW) contaminants from bone gelatin to extract high-molecular-weight (HMW) proteins that are derived from original bone collagen, though it cannot remove HMW collagen crosslinked with humic acids. Therefore, ultrafiltration is often used to obtain more accurate {sup 14}C dates of bones. However, ultrafiltration may introduce new contaminants to bone gelatins, mainly from ultrafilters used. To study the effects of ultrafiltration on {sup 14}C age, we analyzed the C/N ratio, {delta}{sup 13}C{sub PDB} and {delta}{sup 15}N{sub AIR} values, and {sup 14}C ages of acid-soluble bone collagen obtained by decalcification, gelatin extracted from acid-insoluble bone collagen, and the HMW gelatin and LMW fractions produced during ultrafiltration of the extracted gelatin. Bone samples from the Fifth International Radiocarbon Intercomparison (VIRI) were used: VIRI-E (mammoth), -F (horse), -G (human), and -I (whale). In this study, carbon and nitrogen content and gelatin yields were used to evaluate collagen preservation in the VIRI bone samples. Radiocarbon ages, {delta}{sup 13}C{sub PDB} and {delta}{sup 15}N{sub AIR} values of unfiltered and HMW gelatins were obtained and compared with the published consensus values. The LMW fraction was found to exhibit different values from those of the other fractions, indicating the possible presence of extraneous contamination. The Vivaspin Trade-Mark-Sign 6 ultrafilters used in this study were analyzed and radiocarbon dated both before and after cleaning. We present evidence to suggest that LMW fraction contaminants could be derived from the ultrafilters rather than humic substances. Excessively long ultrafiltration time was suspected to have contaminated the bone samples with material from the ultrafilter, because those samples exhibited older {sup 14}C ages than did those filtered for shorter durations. The results in this study indicate that {sup 14}C ages of unfiltered

Periodontal tissue regeneration with PRP incorporated gelatin hydrogel sponges

International Nuclear Information System (INIS)

Nakajima, Dai; Tabata, Yasuhiko; Sato, Soh

2015-01-01

Gelatin hydrogels have been designed and prepared for the controlled release of the transforming growth factor (TGF-b1) and the platelet-derived growth factor (PDGF-BB). PRP (Platelet rich plasma) contains many growth factors including the PDGF and TGF-b1. The objective of this study was to evaluate the regeneration of periodontal tissue following the controlled release of growth factors in PRP. For the periodontal ligament cells and osteoblast, PRP of different concentrations was added. The assessment of DNA, mitochondrial activity and ALP activity were measured. To evaluate the TGF-β1 release from PRP incorporated gelatin sponge, amounts of TGF-β1 in each supernatant sample were determined by the ELISA. Transplantation experiments to prepare a bone defect in a rat alveolar bone were an implanted gelatin sponge incorporated with different concentration PRP. In DNA assay and MTT assay, after the addition of PRP to the periodontal ligament cells and osteoblast, the cell count and mitochondrial activity had increased the most in the group with the addition of 5  ×  PRP. In the ALP assay, after the addition of PRP to the periodontal ligament cells, the cell activity had increased the most in the group with the addition of 3  ×  PRP. In the transplantation, the size of the bone regenerated in the defect with 3  ×  PRP incorporated gelatin sponge was larger than that of the other group. (paper)

Periodontal tissue regeneration with PRP incorporated gelatin hydrogel sponges.

Science.gov (United States)

Nakajima, Dai; Tabata, Yasuhiko; Sato, Soh

2015-10-20

Gelatin hydrogels have been designed and prepared for the controlled release of the transforming growth factor (TGF-b1) and the platelet-derived growth factor (PDGF-BB). PRP (Platelet rich plasma) contains many growth factors including the PDGF and TGF-b1. The objective of this study was to evaluate the regeneration of periodontal tissue following the controlled release of growth factors in PRP. For the periodontal ligament cells and osteoblast, PRP of different concentrations was added. The assessment of DNA, mitochondrial activity and ALP activity were measured. To evaluate the TGF-β1 release from PRP incorporated gelatin sponge, amounts of TGF-β1 in each supernatant sample were determined by the ELISA. Transplantation experiments to prepare a bone defect in a rat alveolar bone were an implanted gelatin sponge incorporated with different concentration PRP. In DNA assay and MTT assay, after the addition of PRP to the periodontal ligament cells and osteoblast, the cell count and mitochondrial activity had increased the most in the group with the addition of 5  ×  PRP. In the ALP assay, after the addition of PRP to the periodontal ligament cells, the cell activity had increased the most in the group with the addition of 3  ×  PRP. In the transplantation, the size of the bone regenerated in the defect with 3  ×  PRP incorporated gelatin sponge was larger than that of the other group.

BONE REGENERATION AFTER DEMINERALIZED BONE MATRIX AND CASTOR OIL (RICINUS COMMUNIS) POLYURETHANE IMPLANTATION

Science.gov (United States)

Leite, Fábio Renato Manzolli; Ramalho, Lizeti Toledo de Oliveira

2008-01-01

Innocuous biocompatible materials have been searched to repair or reconstruct bone defects. Their goal is to restore the function of live or dead tissues. This study compared connective tissue and bone reaction when exposed to demineralized bovine bone matrix and a polyurethane resin derived from castor bean (Ricinus communis). Forty-five rats were assigned to 3 groups of 15 animals (control, bovine bone and polyurethane). A cylindrical defect was created on mandible base and filled with bovine bone matrix and the polyurethane. Control group received no treatment. Analyses were performed after 15, 45 and 60 days (5 animals each). Histological analysis revealed connective tissue tolerance to bovine bone with local inflammatory response similar to that of the control group. After 15 days, all groups demonstrated similar outcomes, with mild inflammatory reaction, probably due to the surgical procedure rather than to the material. In the polymer group, after 60 days, scarce multinucleated cells could still be observed. In general, all groups showed good stability and osteogenic connective tissue with blood vessels into the surgical area. The results suggest biocompatibility of both materials, seen by their integration into rat mandible. Moreover, the polyurethane seems to be an alternative in bone reconstruction and it is an inexhaustible source of biomaterial. PMID:19089203

Trisacryl Gelatin Microspheres Versus Polyvinyl Alcohol Particles in the Preoperative Embolization of Bone Neoplasms

International Nuclear Information System (INIS)

Basile, Antonio; Rand, Thomas; Lomoschitz, Fritz; Toma, Cyril; Lupattelli, Tommaso; Kettenbach, Joachim; Lammer, Johannes

2004-01-01

The aim of this study was to compare the efficacy of trisacryl gelatin microspheres versus polyvinyl alcohol particles (PVA) in the preoperative embolization of bone neoplasms, on the basis of intraoperative blood loss quantified by the differences in preoperative and postoperative hematic levels of hemoglobin, hematocrit and erythrocytes count. From January 1997 to December 2002, preoperative embolization of bone tumors (either primary or secondary) was carried out in 49 patients (age range 12/78), 20 of whom were treated with trysacril gelatin microspheres (group A) and 29 with PVA particles (group B). The delay between embolization and surgery ranged from 1 to 13 days in group A and 1 to 4 days in group B. As used in international protocols, we considered hematic levels of hemoglobin, hematocrit and erythrocytes count for the measurement of intraoperative blood loss then the differences in pre- and postoperative levels were used as statistical comparative parameters. We compared the values of patients treated with embospheres (n = 10) and PVA (n = 18) alone, and patients treated with (group A = 10; group B = 11) versus patients treated without other additional embolic materials in each group (group A = 10; group B = 18). According to the Student's t-test (p < 0.05), the difference of hematic parameters between patients treated by embospheres and PVA alone were significant; otherwise there was no significant difference between patients treated with only one embolic material (embospheres and PVA) versus those treated with other additional embolic agents in each group. The patients treated with microspheres had a minor quantification of intraoperative blood loss compared to those who received PVA particles. Furthermore, they had a minor increase of bleeding related to the delay time between embolization and surgery. The use of additional embolic material did not improve the efficacy of the procedure in either group of patients

Local effect of zoledronic acid on new bone formation in posterolateral spinal fusion with demineralized bone matrix in a murine model.

Science.gov (United States)

Zwolak, Pawel; Farei-Campagna, Jan; Jentzsch, Thorsten; von Rechenberg, Brigitte; Werner, Clément M

2018-01-01

Posterolateral spinal fusion is a common orthopaedic surgery performed to treat degenerative and traumatic deformities of the spinal column. In posteriolateral spinal fusion, different osteoinductive demineralized bone matrix products have been previously investigated. We evaluated the effect of locally applied zoledronic acid in combination with commercially available demineralized bone matrix putty on new bone formation in posterolateral spinal fusion in a murine in vivo model. A posterolateral sacral spine fusion in murine model was used to evaluate the new bone formation. We used the sacral spine fusion model to model the clinical situation in which a bone graft or demineralized bone matrix is applied after dorsal instrumentation of the spine. In our study, group 1 received decortications only (n = 10), group 2 received decortication, and absorbable collagen sponge carrier, group 3 received decortication and absorbable collagen sponge carrier with zoledronic acid in dose 10 µg, group 4 received demineralized bone matrix putty (DBM putty) plus decortication (n = 10), and group 5 received DBM putty, decortication and locally applied zoledronic acid in dose 10 µg. Imaging was performed using MicroCT for new bone formation assessment. Also, murine spines were harvested for histopathological analysis 10 weeks after surgery. The surgery performed through midline posterior approach was reproducible. In group with decortication alone there was no new bone formation. Application of demineralized bone matrix putty alone produced new bone formation which bridged the S1-S4 laminae. Local application of zoledronic acid to demineralized bone matrix putty resulted in significant increase of new bone formation as compared to demineralized bone matrix putty group alone. A single local application of zoledronic acid with DBM putty during posterolateral fusion in sacral murine spine model increased significantly new bone formation in situ in our model. Therefore, our

Gelatin- and starch-based hydrogels. Part A: Hydrogel development, characterization and coating.

Science.gov (United States)

Van Nieuwenhove, Ine; Salamon, Achim; Peters, Kirsten; Graulus, Geert-Jan; Martins, José C; Frankel, Daniel; Kersemans, Ken; De Vos, Filip; Van Vlierberghe, Sandra; Dubruel, Peter

2016-11-05

The present work aims at constructing the ideal scaffold matrix of which the physico-chemical properties can be altered according to the targeted tissue regeneration application. Ideally, this scaffold should resemble the natural extracellular matrix (ECM) as close as possible both in terms of chemical composition and mechanical properties. Therefore, hydrogel films were developed consisting of methacrylamide-modified gelatin and starch-pentenoate building blocks because the ECM can be considered as a crosslinked hydrogel network consisting of both polysaccharides and structural, signaling and cell-adhesive proteins. For the gelatin hydrogels, three different substitution degrees were evaluated including 31%, 72% and 95%. A substitution degree of 32% was applied for the starch-pentenoate building block. Pure gelatin hydrogels films as well as interpenetrating networks with gelatin and starch were developed. Subsequently, these films were characterized using gel fraction and swelling experiments, high resolution-magic angle spinning (1)H NMR spectroscopy, rheology, infrared mapping and atomic force microscopy. The results indicate that both the mechanical properties and the swelling extent of the developed hydrogel films can be controlled by varying the chemical composition and the degree of substitution of the methacrylamide-modified gelatin applied. The storage moduli of the developed materials ranged between 14 and 63kPa. Phase separation was observed for the IPNs for which separated starch domains could be distinguished located in the surrounding gelatin matrix. Furthermore, we evaluated the affinity of aggrecan for gelatin by atomic force microscopy and radiolabeling experiments. We found that aggrecan can be applied as a bioactive coating for gelatin hydrogels by a straightforward physisorption procedure. Thus, we achieved distinct fine-tuning of the physico-chemical properties of these hydrogels which render them promising candidates for tissue engineering

Determination of the shear modulus of gelatine hydrogels by magnetization measurements using dispersed nickel nanorods as mechanical probes

International Nuclear Information System (INIS)

Bender, P.; Tschöpe, A.; Birringer, R.

2013-01-01

Ni nanorods are dispersed into gelatine gels and used as nanoprobes to estimate the shear modulus of the surrounding gel matrix by magnetization measurements. The nanorods are synthesized via pulsed electrodeposition of Ni into porous alumina, released from the templates by dissolution of the oxide layer and after several processing steps dispersed into gelatine gels with an isotropic orientation-distribution. Magnetization measurements of the resulting gels show a significant influence of the gelatine concentration on their magnetic behavior. In particular, with decreasing gelatine concentration the measured coercivity is reduced indicating a mechanical rotation of the nanorods in the field direction. A theoretical model which relates the measured coercivity to the shear modulus of the surrounding gel matrix is introduced and applied to investigate the ageing process of gelatine gels with different gelatine concentrations at room temperature. - Highlights: • AAO-template synthesis of uniaxial ferromagnetic single domain Ni nanorods. • Embedding nanorods as magnetic probes in soft elastic gelatine hydrogels. • Coercivity of isotropic samples increases with gelation time and gelatine concentration. • Quantitative relationship between coercivity and matrix shear modulus is obtained from an extended Stoner–Wohlfarth-model. • Semi-quantitative method for magnetic rheometry of soft elastic materials

Determination of the shear modulus of gelatine hydrogels by magnetization measurements using dispersed nickel nanorods as mechanical probes

Energy Technology Data Exchange (ETDEWEB)

Bender, P., E-mail: nano@p-bender.de; Tschöpe, A., E-mail: antsch@mx.uni-saarland.de; Birringer, R., E-mail: r.birringer@nano.uni-saarland.de

2013-11-15

Ni nanorods are dispersed into gelatine gels and used as nanoprobes to estimate the shear modulus of the surrounding gel matrix by magnetization measurements. The nanorods are synthesized via pulsed electrodeposition of Ni into porous alumina, released from the templates by dissolution of the oxide layer and after several processing steps dispersed into gelatine gels with an isotropic orientation-distribution. Magnetization measurements of the resulting gels show a significant influence of the gelatine concentration on their magnetic behavior. In particular, with decreasing gelatine concentration the measured coercivity is reduced indicating a mechanical rotation of the nanorods in the field direction. A theoretical model which relates the measured coercivity to the shear modulus of the surrounding gel matrix is introduced and applied to investigate the ageing process of gelatine gels with different gelatine concentrations at room temperature. - Highlights: • AAO-template synthesis of uniaxial ferromagnetic single domain Ni nanorods. • Embedding nanorods as magnetic probes in soft elastic gelatine hydrogels. • Coercivity of isotropic samples increases with gelation time and gelatine concentration. • Quantitative relationship between coercivity and matrix shear modulus is obtained from an extended Stoner–Wohlfarth-model. • Semi-quantitative method for magnetic rheometry of soft elastic materials.

In situ localization of gelatinolytic activity in the extracellular matrix of metastases of colon cancer in rat liver using quenched fluorogenic DQ-gelatin

NARCIS (Netherlands)

Mook, Olaf R. F.; van Overbeek, Claudia; Ackema, Eleonora G.; van Maldegem, Febe; Frederiks, Wilma M.

2003-01-01

Matrix metalloproteinases (MMPs) such as gelatinases are believed to play an important role in invasion and metastasis of cancer. In this study we investigated the possible role of MMP-2 and MMP-9 in an experimental model of colon cancer metastasis in rat liver. We demonstrated with gelatin

In vitro study of nano-hydroxyapatite/chitosan–gelatin composites for bio-applications

Directory of Open Access Journals (Sweden)

Khaled R. Mohamed

2014-03-01

Full Text Available The present work aims to study the in vitro properties of nano-hydroxyapatite/chitosan–gelatin composite materials. In vitro behavior was performed in simulated body fluid (SBF to verify the formation of apatite layer onto the composite surfaces. The in vitro data proved the deposition of calcium and phosphorus ions onto hydroxyapatite /polymeric composite surfaces especially those containing high concentrations of polymer content. The degradation of the composites decreased with increase in the polymeric matrix content and highly decreased in the presence of citric acid (CA, especially these composites which contain 30% polymeric content. The water absorption of the composites increased with increase in the polymeric content and highly increased with CA addition. The Fourier transformed infrared reflectance (FT-IR and scanning electron microscope (SEM for the composites confirmed the formation of bone-like apatite layer on the composite surfaces, especially those containing high content of polymers (30% with 0.2 M of CA. These promising composites have suitable properties for bio-applications such as bone grafting and bone tissue engineering applications in the future.

A novel bio-inorganic bone implant containing deglued bone

Indian Academy of Sciences (India)

With the aim of developing an ideal bone graft, a new bone grafting material was developed using deglued bone, chitosan and gelatin. Deglued bone (DGB) which is a by-product of bone glue industries and has the close crystallographic similarities of hydroxyapatite was used as main component in the preparation of bone ...

Bone induction by composite of bioerodible polyorthoester and demineralized bone matrix in rats

DEFF Research Database (Denmark)

Pinholt, E M; Solheim, E; Bang, G

1991-01-01

A composite of a local, sustained, drug-release system, Alzamer bioerodible polyorthoester, and demineralized bone-matrix (DBM) particles implanted in the abdominal muscle of 89 Wistar rats induced cartilage and bone formation at the same rate as DBM when evaluated histologically and by 85Sr uptake....... The composite implant was technically easier to use than DBM alone....

Bone induction by composite of bioerodible polyorthoester and deminiralized bone matrix in rats

International Nuclear Information System (INIS)

Pinholt, E.M.; Solheim, E.; Bang, G.; Sudmann, E.

1991-01-01

A composite of a local, sustained, drug-release system, Alzamer bioerodible polyorthoester, and demineralized bone-matrix (DBM) particles implanted in the abdominal muscle of 89 Wistar rats induced cartilage and bone formation at the same rate as DBM when evaluated histologically and by 85 Sr uptake. The composite implant was technically easier to use than DBM alone. (author)

2-Methoxy-2,4-diphenyl-3(2H)-furanone-labeled gelatin zymography and reverse zymography: a rapid real-time method for quantification of matrix metalloproteinases-2 and -9 and tissue inhibitors of metalloproteinases.

Science.gov (United States)

Min, Danqing; Lyons, James Guy; Jia, Junhong; Lo, Lisa; McLennan, Susan V

2006-02-01

Measurement of matrix metalloproteinases (MMPs) and their specific tissue inhibitors of metalloproteinases (TIMPs) by the techniques of zymography and reverse zymography provide useful information regarding the status of matrix accumulation or breakdown. This report describes the use of 2-methoxy-2,4-diphenyl-3(2H)-furanone (MDPF), a fluorescent compound which can be used to label gelatin as a substrate for detection of the gelatin degrading MMP-2 and -9 by zymography. In addition, a modification of the zymographic technique by addition of excess MMPs enables the use of the MDPF-labeled gelatin substrate for the identification and quantification of TIMPs by reverse zymography. Both systems are real-time sensitive reliable quantification techniques, easily used for measurement of these MMPs and TIMPs in clinical, biological, and tissue culture samples.

Fabrication and characterization of electrospun osteon mimicking scaffolds for bone tissue engineering

Energy Technology Data Exchange (ETDEWEB)

Andric, T. [Virginia Tech-Wake Forest School of Biomedical Engineering and Sciences, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061 (United States); Sampson, A.C. [Chemical Engineering, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061 (United States); Freeman, J.W., E-mail: jwfreeman@vt.edu [Virginia Tech-Wake Forest School of Biomedical Engineering and Sciences, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061 (United States)

2011-01-01

Skeletal loss and bone deficiencies are a major worldwide problem with over 600,000 procedures performed in the US alone annually, making bone one of the most transplanted tissues, second to blood only. Bone is a composite tissue composed of organic matrix, inorganic bone mineral, and water. Structurally bone is organized into two distinct types: trabecular (or cancellous) and cortical (or compact) bones. Trabecular bone is characterized by an extensive interconnected network of pores. Cortical bone is composed of tightly packed units, called osteons, oriented parallel along to the axis of the bone. While the majority of scaffolds attempt to replicate the structure of the trabecular bone, fewer attempts have been made to create scaffolds to mimic the structure of cortical bone. The aim of this study was to develop a technique to fabricate scaffolds that mimic the organization of an osteon, the structural unit of cortical bone. We successfully built a rotating stage for PGA fibers and utilized it for collecting electrospun nanofibers and creating scaffolds. Resulting scaffolds consisted of concentric layers of electrospun PLLA or gelatin/PLLA nanofibers wrapped around PGA microfiber core with diameters that ranged from 200 to 600 {mu}m. Scaffolds were mineralized by incubation in 10x simulated body fluid, and scaffolds composed of 10%gelatin/PLLA had significantly higher amounts of calcium phosphate. The electrospun scaffolds also supported cellular attachment and proliferation of MC3T3 cells over the period of 28 days.

Iota-carrageenan/chitosan/gelatin scaffold for the osteogenic differentiation of adipose-derived MSCs in vitro.

Science.gov (United States)

Li, Junjie; Yang, Boguang; Qian, Yufeng; Wang, Qiyu; Han, Ruijin; Hao, Tong; Shu, Yao; Zhang, Yabin; Yao, Fanglian; Wang, Changyong

2015-10-01

In this study, we have developed ι-carrageenan/chitosan/gelatin (CCG) scaffold containing multiple functional groups (-NH2 , -OH, -COOH, and -SO3 H) to resemble the native extracellular matrix (ECM), using the ion-shielding technology and ultrasonic dispersion method. Fourier transform infrared spectroscopy (FTIR) of the CCG scaffolds suggests that the formation of CCG network involves electrostatic interactions between ι-carrageenan (ι-CA) and chitosan/gelatin, and the covalent cross-linking among amino groups of chitosan and/or gelatin. Scanning electron microscopic (SEM) observation reveals that the porous structure of scaffolds can be modulated by the ratio of ι-CA to chitosan/gelatin. The swelling ratio of the hydrogels increases as the ι-CA contents increase. Using differential scanning calorimetry, we found that the double helix structure of ι-CA is only stabilized at low contents of ι-CA in the CCG scaffolds (e.g., 5 wt %). The scaffolds containing 5% ι-CA showed the best protein adsorption capacity (4.46 ± 0.63 μg protein/mg scaffold) and elastic modulus (5.37 ± 1.03 MPa). In addition, the CCG scaffolds exhibit excellent support for adipose-derived mesenchymal stem cells (ADMSCs) attachment and proliferation, and they can improve the osteogenic differentiation and neovascularization capacities of ADMSCs. Overall, we conclude that the CCG may represent an ideal scaffold material for bone tissue engineering. © 2014 Wiley Periodicals, Inc.

Synthesis and characterization of hydroxyapatite-gelatine composite materials for orthopaedic application

International Nuclear Information System (INIS)

Yanovska, A.; Kuznetsov, V.; Stanislavov, A.; Husak, E.; Pogorielov, M.; Starikov, V.; Bolshanina, S.; Danilchenko, S.

2016-01-01

The composite materials based on hydroxyapatite (HA) and gelatine (Gel) with addition of silver and zirconium oxide were obtained. The study investigates a combination of low powered ultrasonic irradiation and low concentration of gelatine in the co-precipitation synthesis. These composites have different weight ratios of organic/inorganic components and may be synthesized in two ways: simple mixing and co-precipitation. Both of which were compared. The estimation of porosity, in vivo testing, surface morphology and phase composition as well as the IR-analysis were provided. Hydroxyapatite was the main crystalline phase in obtained composites. While around powdered HA-Gel composite the connective tissue capsule is formed without bone tissue formation, HA-Gel-Ag porous composite implantation leads to formation of new bone tissue and activation of cell proliferation. Addition of silver ions into composite material allows decreasing inflammation on the first stage of implantation and has positive effect on bone tissue formation. Some of the obtained composite materials containing silver or ZrO_2 are biocompatible. bio-resorbable and osteoconductive with high level of porosity (75–85%). - Highlights: • Hydroxyapatite-gelatine composites with addition of Ag"+ and ZrO_2 were obtained. • Composites were synthesized in two ways: simple mixing and co-precipitation. • Co-precipitation synthesis combined ultrasonic treatment and low concentration of gelatine. • Obtained composites have different weight ratios of organic/inorganic components. • Some composites are osteoconductive and all of them have high level of porosity (75–85%).

Influence of lyophilization factors and gelatin concentration on pore structures of atelocollagen/gelatin sponge biomaterial.

Science.gov (United States)

Yang, Longqiang; Tanabe, Koji; Miura, Tadashi; Yoshinari, Masao; Takemoto, Shinji; Shintani, Seikou; Kasahara, Masataka

2017-07-26

This study aimed to investigate influences of lyophilization factors and gelatin concentration on pore structures of ACG sponge. ACG sponges of different freezing temperatures (-30, -80 and -196 o C), freezing times (1, 2 and 24 h), gelatin concentrations (0.6%AC+0.15%G, 0.6%AC+0.6%G and 0.6%AC+2.4%G), and with 500 μM fluvastatin were fabricated. Pore structures including porosity and pore size were analyzed by scanning electron microscopy and ImageJ. The cytotoxic effects of ACG sponges were evaluated in vitro. Freezing temperature did not affect porosity while high freezing temperature (-30 o C) increased pore size. The high gelatin concentration group (0.6%AC+2.4%G) had decreased porosity and pore size. Freezing time and 500 μM fluvastatin did not affect pore structures. The cytotoxicity and cell proliferation assays revealed that ACG sponges had no cytotoxic effects on human mesenchymal stromal cell growth and proliferation. These results indicate that ACG sponge may be a good biomaterial scaffold for bone regeneration.

Variation of Hydroxyapatite Content in Soft Gelatin Affects Mesenchymal Stem Cell Differentiation

Directory of Open Access Journals (Sweden)

Fahsai Kantawong

2016-01-01

Full Text Available Gelatin is a common material used in tissue engineering and hydroxyapatite (HA has a composition and structure similar to natural bone mineral. HA is also used to increase the adhesion ability of scaffolds. The physical and mechanical properties of gelatin, together with the chemical properties of HA, can affect cell differentiation. The main purpose of this study is to investigate the gene expression of human mesenchymal stem cells (HMSCs upon culturing on gelatin composite with HA. Low amounts of HA were introduced into the gelatin in order to modulate properties of gelatin. Three types of hydrogel were fabricated by glutaraldehyde crosslinking before lyophilization to produce the porous 3D structure: (1 pure gelatin, (2 0.5 mg/ml HA in gelatin, and (3 1 mg/ml HA in gelatin. The fabricated hydrogels were used as scaffolds to cultivate HMSCs for two periods - 24 hours and 3 weeks. The results showed that all types of fabricated hydrogels could be used to cultivate HMSCs. Changes of gene expressions indicated that the HMSCs cultured on the 1 mg/ml HA in gelatin showed neuronal lineage-specific differentiation.

The sintered microsphere matrix for bone tissue engineering: in vitro osteoconductivity studies.

Science.gov (United States)

Borden, Mark; Attawia, Mohamed; Laurencin, Cato T

2002-09-05

A tissue engineering approach has been used to design three-dimensional synthetic matrices for bone repair. The osteoconductivity and degradation profile of a novel polymeric bone-graft substitute was evaluated in an in vitro setting. Using the copolymer poly(lactide-co-glycolide) [PLAGA], a sintering technique based on microsphere technology was used to fabricate three-dimensional porous scaffolds for bone regeneration. Osteoblasts and fibroblasts were seeded onto a 50:50 PLAGA scaffold. Morphologic evaluation through scanning electron microscopy demonstrated that both cell types attached and spread over the scaffold. Cells migrated through the matrix using cytoplasmic extensions to bridge the structure. Cross-sectional images indicated that cellular proliferation had penetrated into the matrix approximately 700 microm from the surface. Examination of the surfaces of cell/matrix constructs demonstrated that cellular proliferation had encompassed the pores of the matrix by 14 days of cell culture. With the aim of optimizing polymer composition and polymer molecular weight, a degradation study was conducted utilizing the matrix. The results demonstrate that degradation of the sintered matrix is dependent on molecular weight, copolymer ratio, and pore volume. From this data, it was determined that 75:25 PLAGA with an initial molecular weight of 100,000 has an optimal degradation profile. These studies show that the sintered microsphere matrix has an osteoconductive structure capable of functioning as a cellular scaffold with a degradation profile suitable for bone regeneration. Copyright 2002 Wiley Periodicals, Inc.

Electron beam effects on gelatin polymer

Energy Technology Data Exchange (ETDEWEB)

Inamura, Patricia Y.; Shimazaki, Kleber; Souza, Clecia de M.; Moura, Esperidiana A.B.; Mastro, Nelida L. del, E-mail: patyoko@yahoo.co [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil); Colombo, Maria A., E-mail: mascolombo@yahoo.com.b [Faculdade de Tecnologia da Zona Leste, Sao Paulo, SP (Brazil)

2009-07-01

The main field of electron-beam radiation processing applications is the modification of polymeric material. Polymer development includes new pathways to produce natural polymers with better mechanical and barrier properties and thermal stability. The aim of this paper was to investigate the behavior of a gelatin/acrylamide polymer treated by electron-beam radiation. Gelatin is a heterogeneous mixture of water-soluble proteins of high average molecular mass derived by hydrolytic action from animal collagen, a fibrous insoluble protein, which is widely found in nature as the major constituent of skin, bones and connective tissue. Hydrolyzed collagen is composed of a unique sequence of amino acids, characterized particularly by the high content of glycine, proline and hydroxyproline. Among biomaterials, gelatin is an interesting material because is a partially crystalline polymer and has a relatively low melting point. Samples of gelatin together with glycerin as plasticizer and acrylamide as copolymer were irradiated with doses of 10 kGy and 40 kGy, using an electron beam accelerator, dose rate 22.41kGy/s, at room temperature in presence of air. After irradiation, some preliminary analyses were done like viscometry, texture analyses and colorimetry. The results of the diverse tests showed changes that can be ascribed to radiation-induced crosslinking. The electron-beam processed acrylamide-gelatin polymer using glycerin as plasticizer must be first extensively characterized before to be used for general applications. (author)

Electron beam effects on gelatin polymer

International Nuclear Information System (INIS)

Inamura, Patricia Y.; Shimazaki, Kleber; Souza, Clecia de M.; Moura, Esperidiana A.B.; Mastro, Nelida L. del; Colombo, Maria A.

2009-01-01

The main field of electron-beam radiation processing applications is the modification of polymeric material. Polymer development includes new pathways to produce natural polymers with better mechanical and barrier properties and thermal stability. The aim of this paper was to investigate the behavior of a gelatin/acrylamide polymer treated by electron-beam radiation. Gelatin is a heterogeneous mixture of water-soluble proteins of high average molecular mass derived by hydrolytic action from animal collagen, a fibrous insoluble protein, which is widely found in nature as the major constituent of skin, bones and connective tissue. Hydrolyzed collagen is composed of a unique sequence of amino acids, characterized particularly by the high content of glycine, proline and hydroxyproline. Among biomaterials, gelatin is an interesting material because is a partially crystalline polymer and has a relatively low melting point. Samples of gelatin together with glycerin as plasticizer and acrylamide as copolymer were irradiated with doses of 10 kGy and 40 kGy, using an electron beam accelerator, dose rate 22.41kGy/s, at room temperature in presence of air. After irradiation, some preliminary analyses were done like viscometry, texture analyses and colorimetry. The results of the diverse tests showed changes that can be ascribed to radiation-induced crosslinking. The electron-beam processed acrylamide-gelatin polymer using glycerin as plasticizer must be first extensively characterized before to be used for general applications. (author)

Differences in cytocompatibility between collagen, gelatin and keratin

Energy Technology Data Exchange (ETDEWEB)

Wang, Yanfang; Zhang, Weiwei; Yuan, Jiang, E-mail: jyuan@njnu.edu.cn; Shen, Jian, E-mail: jshen@njnu.edu.cn

2016-02-01

Keratins are cysteine-rich intermediate filament proteins found in the cytoskeleton of the epithelial cells and in the matrix of hair, feathers, wool, nails and horns. The natural abundance of cell adhesion sequences, RGD (Arg-Gly-Asp) and LDV (Leu-Asp-Val), makes them suitable for tissue engineering applications. The purpose of our study is to evaluate their cytocompatibility as compared to well-known collagen and gelatin proteins. Herein, collagen, gelatin and keratin were blended with poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and electrospun to afford nanofibrous mats, respectively. These PHBV/protein composite mats were characterized by field emission scanning electron microscopy (FE-SEM), attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR), X-ray photoelectron spectroscopy (XPS), and dynamic mechanical analysis (DMA). The cytocompatibility was evaluated with cell adhesion, cell viability and cell proliferation. The data from MTT and BrDU revealed that collagen had significantly superior cytocompatibility as compared to gelatin and keratin. Gelatin showed a better cytocompatibility than keratin without statistical significance difference. Finally, we gave the reasons to account for the above conclusions. - Highlights: • Collagen, gelatin and keratin were coelectrospun with PHBV to afford nanofibrous mats. • Cytocompatibility was evaluated with cell adhesion, cell viability and cell proliferation. • Collagen had significantly superior cytocompatibility as compared to gelatin and keratin.

Synthesis and characterization of hydroxyapatite-gelatine composite materials for orthopaedic application

Energy Technology Data Exchange (ETDEWEB)

Yanovska, A., E-mail: biophy@yandex.ru [Institute of Applied Physics, National Academy of Sciences of Ukraine, 58 Petropavlovskaya Str., 40000, Sumy (Ukraine); Sumy State University, Ministry of Education and Science of Ukraine, 2 R. Korsakova Str., 40007, Sumy (Ukraine); Kuznetsov, V. [Institute of Applied Physics, National Academy of Sciences of Ukraine, 58 Petropavlovskaya Str., 40000, Sumy (Ukraine); Sumy State University, Ministry of Education and Science of Ukraine, 2 R. Korsakova Str., 40007, Sumy (Ukraine); Stanislavov, A. [Institute of Applied Physics, National Academy of Sciences of Ukraine, 58 Petropavlovskaya Str., 40000, Sumy (Ukraine); Husak, E. [Institute of Applied Physics, National Academy of Sciences of Ukraine, 58 Petropavlovskaya Str., 40000, Sumy (Ukraine); Sumy State University, Ministry of Education and Science of Ukraine, 2 R. Korsakova Str., 40007, Sumy (Ukraine); Pogorielov, M. [Sumy State University, Ministry of Education and Science of Ukraine, 2 R. Korsakova Str., 40007, Sumy (Ukraine); Starikov, V. [National Technical University ”Kharkov Polytechnic Institute”, 21 Frunze Str., 61002, Kharkov (Ukraine); Bolshanina, S. [Sumy State University, Ministry of Education and Science of Ukraine, 2 R. Korsakova Str., 40007, Sumy (Ukraine); Danilchenko, S. [Institute of Applied Physics, National Academy of Sciences of Ukraine, 58 Petropavlovskaya Str., 40000, Sumy (Ukraine)

2016-11-01

The composite materials based on hydroxyapatite (HA) and gelatine (Gel) with addition of silver and zirconium oxide were obtained. The study investigates a combination of low powered ultrasonic irradiation and low concentration of gelatine in the co-precipitation synthesis. These composites have different weight ratios of organic/inorganic components and may be synthesized in two ways: simple mixing and co-precipitation. Both of which were compared. The estimation of porosity, in vivo testing, surface morphology and phase composition as well as the IR-analysis were provided. Hydroxyapatite was the main crystalline phase in obtained composites. While around powdered HA-Gel composite the connective tissue capsule is formed without bone tissue formation, HA-Gel-Ag porous composite implantation leads to formation of new bone tissue and activation of cell proliferation. Addition of silver ions into composite material allows decreasing inflammation on the first stage of implantation and has positive effect on bone tissue formation. Some of the obtained composite materials containing silver or ZrO{sub 2} are biocompatible. bio-resorbable and osteoconductive with high level of porosity (75–85%). - Highlights: • Hydroxyapatite-gelatine composites with addition of Ag{sup +} and ZrO{sub 2} were obtained. • Composites were synthesized in two ways: simple mixing and co-precipitation. • Co-precipitation synthesis combined ultrasonic treatment and low concentration of gelatine. • Obtained composites have different weight ratios of organic/inorganic components. • Some composites are osteoconductive and all of them have high level of porosity (75–85%).

Preparation of Rhodamine B Fluorescent Poly(methacrylic acid Coated Gelatin Nanoparticles

Directory of Open Access Journals (Sweden)

Zhenhai Gan

2011-01-01

Full Text Available Poly(methacrylic acid (PMAA-coated gelatin nanoparticles encapsulated with fluorescent dye rhodamine B were prepared by the coacervation method with the aim to retard the release of rhodamine B from the gelatin matrix. With sodium sulfate as coacervation reagent for gelatin, a kind of biopolymer with excellent biocompatibility, the formed gelatin nanoparticles were cross-linked by formaldehyde followed by the polymerization of methacrylic acid coating. The fluorescent poly(methacrylic acid coated gelatin (FPMAAG nanoparticles had a uniform spherical shape and a size distribution of 60±5 nm. Infrared spectral analysis confirmed the formation of PMAA coating on the gelatin nanoparticles. Based on UV-Vis spectra, the loading efficiency of rhodamine B for the FPMAAG nanoparticles was 0.26 μg per mg nanoparticles. The encapsulated rhodamine B could sustain for two weeks. Favorable fluorescence property and fluorescence imaging of cells confirmed that the FPMAAG nanoparticles have promising biochemical, bioanalytical, and biomedical applications.

Anabolic action of parathyroid hormone (PTH) does not compromise bone matrix mineral composition or maturation.

Science.gov (United States)

Vrahnas, Christina; Pearson, Thomas A; Brunt, Athena R; Forwood, Mark R; Bambery, Keith R; Tobin, Mark J; Martin, T John; Sims, Natalie A

2016-12-01

Intermittent administration of parathyroid hormone (PTH) is used to stimulate bone formation in patients with osteoporosis. A reduction in the degree of matrix mineralisation has been reported during treatment, which may reflect either production of undermineralised matrix or a greater proportion of new matrix within the bone samples assessed. To explore these alternatives, high resolution synchrotron-based Fourier Transform Infrared Microspectroscopy (sFTIRM) coupled with calcein labelling was used in a region of non-remodelling cortical bone to determine bone composition during anabolic PTH treatment compared with region-matched samples from controls. 8week old male C57BL/6 mice were treated with vehicle or 50μg/kg PTH, 5 times/week for 4weeks (n=7-9/group). Histomorphometry confirmed greater trabecular and periosteal bone formation and 3-point bending tests confirmed greater femoral strength in PTH-treated mice. Dual calcein labels were used to match bone regions by time-since-mineralisation (bone age) and composition was measured by sFTIRM in six 15μm 2 regions at increasing depth perpendicular to the most immature bone on the medial periosteal edge; this allowed in situ measurement of progressive changes in bone matrix during its maturation. The sFTIRM method was validated in vehicle-treated bones where the expected progressive increases in mineral:matrix ratio and collagen crosslink type ratio were detected with increasing bone maturity. We also observed a gradual increase in carbonate content that strongly correlated with an increase in longitudinal stretch of the collagen triple helix (amide I:amide II ratio). PTH treatment did not alter the progressive changes in any of these parameters from the periosteal edge through to the more mature bone. These data provide new information about how the bone matrix matures in situ and confirm that bone deposited during PTH treatment undergoes normal collagen maturation and normal mineral accrual. Copyright © 2016

Angiographic findings of demineralised bone matrix grafts in femoral fracture repair in canines*

International Nuclear Information System (INIS)

Kumar, R.V.S.; Ramakrishna, O.

2000-01-01

Demineralised bone matrix implants and autogenous cancellous bone grafts were evaluated in femoral fracture repair with bone loss in canines, and compared to untreated animals. Angiograms were taken at 3,6,9 and 12 weeks post- operatively using conrary-420 as contrast medium. Early angiograms showed leakage of contrast medium into soft tissue. Grafted groups showed slight hypervascularity and enhanced capillary network. At 12 Weeks the same groups observed nearly normal persistent vascular supply. Early establishment of cortices continuity and draining of contrast medium via extraosseous veins was observed in demineralized bone matrix implanted groups

Bisphosphonate treatment affects trabecular bone apparent modulus through micro-architecture rather than matrix properties

DEFF Research Database (Denmark)

Ding, Ming

2004-01-01

and trabecular architecture independently. Conventional histomorphometry and microdamage data were obtained from the second and third lumbar vertebrae of the same dogs [Bone 28 (2001) 524]. Bisphosphonate treatment resulted in an increased apparent Young's modulus, decreased bone turnover, increased calcified...... matrix density, and increased microdamage. We could not detect any change in the effective Young's modulus of the calcified matrix in the bisphosphonate treated groups. The observed increase in apparent Young's modulus was due to increased bone mass and altered trabecular architecture rather than changes...... in the calcified matrix modulus. We hypothesize that the expected increase in the Young's modulus of the calcified matrix due to the increased calcified matrix density was counteracted by the accumulation of microdamage. Udgivelsesdato: 2004 May...

Experimental Evaluation of the Effectiveness of Demineralized Bone Matrix and Collagenated Heterologous Bone Grafts Used Alone or in Combination with Platelet-Rich Fibrin on Bone Healing in Sinus Floor Augmentation.

Science.gov (United States)

Peker, Elif; Karaca, Inci Rana; Yildirim, Benay

2016-01-01

The aim of this study was an experimental evaluation of the effectiveness of demineralized bone matrix (DBM) and collagenated heterologous bone graft (CHBG) used alone or in combination with platelet-rich fibrin on bone healing in sinus floor augmentation procedures. In this study, 36 New Zealand rabbits were used. The bilateral sinus elevation was performed, and 72 defects were obtained. The rabbit maxillary sinuses were divided into four groups according to the augmentation biomaterials obtained: demineralized bone matrix (Grafton DBM Putty, Osteotech; DBM group), DBM combined with platelet-rich fibrin (PRF; DBM + PRF group), collagenated heterologous bone graft (CHBG; Apatos Mix, OsteoBiol, Tecnoss; CHBG group), CHBG combined with PRF (CHBG + PRF group). All groups were sacrificed at 2, 4, and 8 weeks after surgery for histologic, histomorphometric, and immunohistochemical analyses. The inflammatory reaction was moderate to intense at the second week in all groups and declined from 2 to 8 weeks. New bone formation was started at the second week and increased from 2 to 8 weeks in all groups. There was no significant difference in bone formation between the experimental groups that used PRF mixed graft material and control groups that used only graft material. The percentage of new bone formation showed a significant difference in DBM groups and DBM + PRF groups compared with other groups. There were osteoclasts around all the bone graft materials used, but the percentage of residual graft particles was significantly higher in CHBG groups and CHBG + PRF groups at the eighth week. There is no beneficial effect of the application of PRF in combination with demineralized bone matrix or collagenated heterologous bone graft on bone formation in sinus floor augmentation. The results of this study showed that both collagenated heterologous bone graft and demineralized bone matrix have osteoconductive properties, but demineralized bone matrix showed more bone formation

A Comparative Analysis of Recombinant Human Bone Morphogenetic Protein-2 with a Demineralized Bone Matrix versus Iliac Crest Bone Graft for Secondary Alveolar Bone Grafts in Patients with Cleft Lip and Palate: Review of 501 Cases.

Science.gov (United States)

Hammoudeh, Jeffrey A; Fahradyan, Artur; Gould, Daniel J; Liang, Fan; Imahiyerobo, Thomas; Urbinelli, Leo; Nguyen, JoAnna T; Magee, William; Yen, Stephen; Urata, Mark M

2017-08-01

Alveolar cleft reconstruction using iliac crest bone graft is considered standard of care for children with complete cleft lip and palate at the time of mixed dentition. Harvesting bone may result in donor-site morbidity and additional operating time and length of hospitalization. Recombinant human bone morphogenetic protein (rhBMP)-2 with a demineralized bone matrix is an alternative bone source for alveolar cleft reconstruction. The authors investigated the outcomes of rhBMP-2/demineralized bone matrix versus iliac crest bone graft for alveolar cleft reconstruction by reviewing postoperative surgical complications and cleft closure. A retrospective chart review was conducted for 258 rhBMP-2/demineralized bone matrix procedures (mean follow-up, 2.9 years) and 243 iliac crest bone graft procedures (mean follow-up, 4.1 years) on 414 patients over a 12-year period. The authors compared complications, canine eruption, and alveolar cleft closure between the two groups. In the rhBMP-2/demineralized bone matrix group, one patient required prolonged intubation because of intraoperative airway swelling not thought to be caused by rhBMP-2, 36 reported facial swelling and one required outpatient steroids as treatment, and 12 had dehiscence; however, half of these complications resolved without intervention. Twenty-three of the 228 rhBMP-2/demineralized bone matrix patients and 28 of the 242 iliac crest bone graft patients required repeated surgery for alveolar cleft repair. Findings for canine tooth eruption into the cleft site through the graft were similar between the groups. The rhBMP-2/demineralized bone matrix appears to be an acceptable alternative for alveolar cleft repair. The authors found no increase in serious adverse events with the use of this material. Local complications, such as swelling and minor wound dehiscence, predominantly improved without intervention. Therapeutic, III.

Mussel-inspired graphene oxide nanosheet-enwrapped Ti scaffolds with drug-encapsulated gelatin microspheres for bone regeneration.

Science.gov (United States)

Han, Lu; Sun, Honglong; Tang, Pengfei; Li, Pengfei; Xie, Chaoming; Wang, Menghao; Wang, Kefeng; Weng, Jie; Tan, Hui; Ren, Fuzeng; Lu, Xiong

2018-02-27

Graphene oxide (GO) attracts considerable attention for biomedical applications owing to its unique nanostructure and remarkable physicochemical characteristics. However, it is challenging to uniformly deposit GO on chemically inert Ti scaffolds, which have good biocompatibility and wide applications in bone engineering. In this study, a GO-functionalized Ti porous scaffold (GO/Ti scaffold) was prepared by depositing GO onto polydopamine (PDA) modified Ti scaffolds. The mussel-inspired PDA modification facilitated the interaction between GO and Ti surfaces, leading to a uniform coverage of GO on Ti scaffolds. BMP2 and vancomycin (Van) were separately encapsulated into gelatin microspheres (GelMS). Then, drug-containing GelMS were assembled on GO/Ti scaffolds and anchored by the functional groups of GO. The modified scaffold independently delivered multiple biomolecules with different physiochemical properties, without interfering with each other. Thus, the GO/Ti scaffold has the dual functions of inducing bone regeneration and preventing bacterial infection. In summary, this mussel-inspired GO/Ti hybrid scaffold combined the good mechanical properties of Ti scaffolds and the advantages of GO nanosheets. GO nanosheets with their unique nanostructure and functional groups, together with GelMS on Ti scaffolds, are suitable carriers for drug delivery and provide adhesive sites for cell adhesion and create nanostructured environments for bone regeneration.

Tendon Reattachment to Bone in an Ovine Tendon Defect Model of Retraction Using Allogenic and Xenogenic Demineralised Bone Matrix Incorporated with Mesenchymal Stem Cells.

Directory of Open Access Journals (Sweden)

Tanujan Thangarajah

Full Text Available Tendon-bone healing following rotator cuff repairs is mainly impaired by poor tissue quality. Demineralised bone matrix promotes healing of the tendon-bone interface but its role in the treatment of tendon tears with retraction has not been investigated. We hypothesized that cortical demineralised bone matrix used with minimally manipulated mesenchymal stem cells will result in improved function and restoration of the tendon-bone interface with no difference between xenogenic and allogenic scaffolds.In an ovine model, the patellar tendon was detached from the tibial tuberosity and a complete distal tendon transverse defect measuring 1 cm was created. Suture anchors were used to reattach the tendon and xenogenic demineralised bone matrix + minimally manipulated mesenchymal stem cells (n = 5, or allogenic demineralised bone matrix + minimally manipulated mesenchymal stem cells (n = 5 were used to bridge the defect. Graft incorporation into the tendon and its effect on regeneration of the enthesis was assessed using histomorphometry. Force plate analysis was used to assess functional recovery.Compared to the xenograft, the allograft was associated with significantly higher functional weight bearing at 6 (P = 0.047, 9 (P = 0.028, and 12 weeks (P = 0.009. In the allogenic group this was accompanied by greater remodeling of the demineralised bone matrix into tendon-like tissue in the region of the defect (p = 0.015, and a more direct type of enthesis characterized by significantly more fibrocartilage (p = 0.039. No failures of tendon-bone healing were noted in either group.Demineralised bone matrix used with minimally manipulated mesenchymal stem cells promotes healing of the tendon-bone interface in an ovine model of acute tendon retraction, with superior mechanical and histological results associated with use of an allograft.

Function of Matrix IGF-1 in Coupling Bone Resorption and Formation

Science.gov (United States)

Crane, Janet L.; Cao, Xu

2013-01-01

Balancing bone resorption and formation is the quintessential component for the prevention of osteoporosis. Signals that determine the recruitment, replication, differentiation, function, and apoptosis of osteoblasts and osteoclasts direct bone remodeling and determine whether bone tissue is gained, lost, or balanced. Therefore understanding the signaling pathways involved in the coupling process will help develop further targets for osteoporosis therapy, by blocking bone resorption or enhancing bone formation in a space and time dependent manner. Insulin-like growth factor type 1 (IGF-1) has long been known to play a role in bone strength. It is one of the most abundant substances in the bone matrix, circulates systemically and is secreted locally, and has a direct relationship with bone mineral density. Recent data has helped further our understanding of the direct role of IGF-1 signaling in coupling bone remodeling which will be discussed in this review. The bone marrow microenvironment plays a critical role in the fate of MSCs and HSCs and thus how IGF-1 interacts with other factors in the microenvironment are equally important. While previous clinical trials with IGF-1 administration have been unsuccessful at enhancing bone formation, advances in basic science studies have provided insight into further mechanisms that should be considered for future trials. Additional basic science studies dissecting the regulation and the function of matrix IGF-1 in modeling and remodeling will continue to provide further insight for future directions for anabolic therapies for osteoporosis. PMID:24068256

Function of matrix IGF-1 in coupling bone resorption and formation.

Science.gov (United States)

Crane, Janet L; Cao, Xu

2014-02-01

Balancing bone resorption and formation is the quintessential component for the prevention of osteoporosis. Signals that determine the recruitment, replication, differentiation, function, and apoptosis of osteoblasts and osteoclasts direct bone remodeling and determine whether bone tissue is gained, lost, or balanced. Therefore, understanding the signaling pathways involved in the coupling process will help develop further targets for osteoporosis therapy, by blocking bone resorption or enhancing bone formation in a space- and time-dependent manner. Insulin-like growth factor type 1 (IGF-1) has long been known to play a role in bone strength. It is one of the most abundant substances in the bone matrix, circulates systemically and is secreted locally, and has a direct relationship with bone mineral density. Recent data has helped further our understanding of the direct role of IGF-1 signaling in coupling bone remodeling which will be discussed in this review. The bone marrow microenvironment plays a critical role in the fate of mesenchymal stem cells and hematopoietic stem cells and thus how IGF-1 interacts with other factors in the microenvironment are equally important. While previous clinical trials with IGF-1 administration have been unsuccessful at enhancing bone formation, advances in basic science studies have provided insight into further mechanisms that should be considered for future trials. Additional basic science studies dissecting the regulation and the function of matrix IGF-1 in modeling and remodeling will continue to provide further insight for future directions for anabolic therapies for osteoporosis.

[Levels of bone mineral matrix organization and the mechanisms determining parameters of its formation].

Science.gov (United States)

Avrunin, A S; Tikhilov, R M; Abolin, A B; Shcherbak, I G

2005-01-01

Authors suggest to regard bone mineral matrix as the four-level structure. The first level is represented by an internal structure of a mineral, the second--by mineral morphological structure, the third--by coplanar association of minerals, and the fourth--by macroassociation of minerals in a single complex inside each bone. The most probable mechanisms determining stability of reproduction of mineral matrix parameters on each of these levels are shown. As a result of their functioning, the variants of bone mineral matrix structures are formed that are the programmed reflection of specificity of the given site of organic structures.

Insulin-like growth factor I has independent effects on bone matrix formation and cell replication

International Nuclear Information System (INIS)

Hock, J.M.; Centrella, M.; Canalis, E.

1988-01-01

The effects of insulin-like growth factor-I (IGF-I) and insulin on bone matrix synthesis and bone cell replication were studied in cultured 21-day-old fetal rat calvariae. Histomorphometry techniques were developed to measure the incorporation of [2,3- 3 H]proline and [methyl- 3 H]thymidine into bone matrix and bone cell nuclei, respectively, using autoradiographs of sagittal sections of calvariae cultured with IGF-I, insulin, or vehicle for up to 96 h. To confirm an effect on bone formation, IGF-I was also studied for its effects on [ 3 H]proline incorporation into collagenase-digestible protein (CDP) and noncollagen protein and on [ 3 H]thymidine incorporation into acid-precipitable material (DNA). IGF-I at 10(-9)-10(-7) M significantly increased the rate of bone matrix apposition and CDP after 24 h by 45-50% and increased cell labeling by 8-fold in the osteoprogenitor cell zone, by 4-fold in the osteoblast cell zone, and by 2-fold in the periosteal fibroblast zone. Insulin at 10(-9)-10(-6) M also increased matrix apposition rate and CDP by 40-50%, but increased cell labeling by 2-fold only at a concentration of 10(-7) M or higher and then only in the osteoprogenitor cell zone. When hydroxyurea was added to IGF-I-treated bones, the effects of IGF-I on DNA synthesis were abolished, but the increase in bone matrix apposition induced by IGF-I was only partly diminished. In conclusion, IGF-I stimulates matrix synthesis in calvariae, an effect that is partly, although not completely, dependent on its stimulatory effect on DNA synthesis

Graphene oxide decorated electrospun gelatin nanofibers: Fabrication, properties and applications

Energy Technology Data Exchange (ETDEWEB)

Jalaja, K. [Department of Chemistry, Indian Institute of Space Science and Technology, Valiamala, Thiruvananthapuram, Kerala 695 547 (India); Sreehari, V.S. [Indian Institute of Science Education and Research Bhopal, Bhauri, Madhya Pradesh 462066 (India); Kumar, P.R. Anil [Tissue culture laboratory, Biomedical Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Poojappura, Thiruvananthapuram, Kerala 695 012 (India); Nirmala, R. James, E-mail: nirmala@iist.ac.in [Department of Chemistry, Indian Institute of Space Science and Technology, Valiamala, Thiruvananthapuram, Kerala 695 547 (India)

2016-07-01

Gelatin nanofiber fabricated by electrospinning process is found to mimic the complex structural and functional properties of natural extracellular matrix for tissue regeneration. In order to improve the physico-chemical and biological properties of the nanofibers, graphene oxide is incorporated in the gelatin to form graphene oxide decorated gelatin nanofibers. The current research effort is focussed on the fabrication and evaluation of physico-chemical and biological properties of graphene oxide-gelatin composite nanofibers. The presence of graphene oxide in the nanofibers was established by transmission electron microscopy (TEM). We report the effect of incorporation of graphene oxide on the mechanical, thermal and biological performance of the gelatin nanofibers. The tensile strength of gelatin nanofibers was increased from 8.29 ± 0.53 MPa to 21 ± 2.03 MPa after the incorporation of GO. In order to improve the water resistance of nanofibers, natural based cross-linking agent, namely, dextran aldehyde was employed. The cross-linked composite nanofibers showed further increase in the tensile strength up to 56.4 ± 2.03 MPa. Graphene oxide incorporated gelatin nanofibers are evaluated for bacterial activity against gram positive (Staphylococcus aureus) and gram negative (Escherichia coli) bacteria and cyto compatibility using mouse fibroblast cells (L-929 cells). The results indicate that the graphene oxide incorporated gelatin nanofibers do not prevent bacterial growth, nevertheless support the L-929 cell adhesion and proliferation. - Highlights: • Graphene oxide nano reinforced gelatin nanofibers are fabricated by electrospinning. • Graphene oxide (0.5%) loading resulted in increased tensile strength. • GO/gelatin nanofibers are cross-linked with dextran aldehyde. • Composite nanofibers favoured adhesion of L-929 cells. • GO/gelatin mats do not prevent bacterial growth.

Recombinant Gelatin Microspheres : Novel Formulations for Tissue Repair?

NARCIS (Netherlands)

Tuin, Annemarie; Kluijtmans, Sebastiaan G.; Bouwstra, Jan B.; Harmsen, Martin C.; Van Luyn, Marja J. A.

Microspheres (MSs) can function as multifunctional scaffolds in different approaches of tissue repair (TR), as a filler, a slow-release depot for growth factors, or a delivery vehicle for cells. Natural cell adhesion-supporting extracellular matrix components like gelatin are good materials for

Transplantation of artificial gelatin-co-bletillastriata gelatin/ Salvia ...

African Journals Online (AJOL)

Purpose: To evaluate the growth of the composite corium (constructed with fibroblast cells and gelatinco- Bletillastriata gelatin/Salvia miltiorrhiza materials) on rats. Methods: The composite artificial corium was constructed by culturing fibroblast cells in gelatin-co- Bletillastriata gelatin/Salvia miltiorrhiza materials.

Minimally invasive esthetic ridge preservation with growth-factor enhanced bone matrix.

Science.gov (United States)

Nevins, Marc L; Said, Sherif

2017-12-28

Extraction socket preservation procedures are critical to successful esthetic implant therapy. Conventional surgical approaches are technique sensitive and often result in alteration of the soft tissue architecture, which then requires additional corrective surgical procedures. This case series report presents the ability of flapless surgical techniques combined with a growth factor-enhanced bone matrix to provide esthetic ridge preservation at the time of extraction for compromised sockets. When considering esthetic dental implant therapy, preservation, or further enhancement of the available tissue support at the time of tooth extraction may provide an improved esthetic outcome with reduced postoperative sequelae and decreased treatment duration. Advances in minimally invasive surgical techniques combined with recombinant growth factor technology offer an alternative for bone reconstruction while maintaining the gingival architecture for enhanced esthetic outcome. The combination of freeze-dried bone allograft (FDBA) and rhPDGF-BB (platelet-derived growth factor-BB) provides a growth-factor enhanced matrix to induce bone and soft tissue healing. The use of a growth-factor enhanced matrix is an option for minimally invasive ridge preservation procedures for sites with advanced bone loss. Further studies including randomized clinical trials are needed to better understand the extent and limits of these procedures. The use of minimally invasive techniques with growth factors for esthetic ridge preservation reduces patient morbidity associated with more invasive approaches and increases the predictability for enhanced patient outcomes. By reducing the need for autogenous bone grafts the use of this technology is favorable for patient acceptance and ease of treatment process for esthetic dental implant therapy. © 2017 Wiley Periodicals, Inc.

Pathologic fracture through a unicameral bone cyst of the pelvis: CT-guided percutaneous curettage, biopsy, and bone matrix injection.

Science.gov (United States)

Tynan, Jennifer R; Schachar, Norman S; Marshall, Geoffrey B; Gray, Robin R

2005-02-01

Unicameral bone cysts of the pelvis are extremely rare. A 19-year old man presented with a pathologic fracture through a pelvic unicameral bone cyst. He was treated with computed tomography-guided percutaneous curettage, biopsy, and demineralized bone matrix injection. Treatment has proven successful in short-term follow-up.

Gel-Based Gelatin Zymography to Examine Matrix Metalloproteinase Activity in Cell Culture.

Science.gov (United States)

Chhabra, Aastha; Rani, Vibha

2018-01-01

Gelatin zymography, first described by Heussen and Dowdle in the 1980s, is a widely used valuable tool in research and diagnostics. The technique identifies gelatinases by the degradation of their preferential substrate as well as by their molecular weight (kDa). We here describe detailed methodology for the detection of pro- and active- forms of both MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in cells using norepinephrine-stimulated H9c2 cardiomyoblasts as model. An easy to follow step-by-step protocol has been carefully written for reliable results. We also suggest an acceptable method for quantification of gelatin zymograms.

Osteoinductivity of gelatin/β-tricalcium phosphate sponges loaded with different concentrations of mesenchymal stem cells and bone morphogenetic protein-2 in an equine bone defect model.

Science.gov (United States)

Seo, Jong-Pil; Tsuzuki, Nao; Haneda, Shingo; Yamada, Kazutaka; Furuoka, Hidefumi; Tabata, Yasuhiko; Sasaki, Naoki

2014-03-01

Fracture is one of the most life-threatening injuries in horses. Fracture repair is often associated with unsatisfactory outcomes and is associated with a high incidence of complications. This study aimed to evaluate the osteogenic effects of gelatin/β-tricalcium phosphate (GT) sponges loaded with different concentrations/ratios of mesenchymal stem cells (MSCs) and bone morphogenetic protein-2 (BMP-2) in an equine bone defect model. Seven thoroughbred horses were used in this study. Eight bone defects were created in the third metatarsal bones of each horse. Then, eight treatments, namely control, GT, GT/M-5, GT/M-6, GT/M-5/B-1, GT/M-5/B-3, GT/M-6/B-1, and GT/M-6/B-3 were applied to the eight different sites in a randomized manner (M-5: 2 × 10(5) MSCs; M-6: 2 × 10(6) MSCs; B-1: 1 μg of BMP-2; B-3: 3 μg of BMP-2). Repair of bone defects was assessed by radiography, quantitative computed tomography (QCT), and histopathological evaluation. Radiographic scores and CT values were significantly lower in the control group than in the other groups, while they were significantly higher in the GT/M-5/B-3 and GT/M-6/B-3 groups than in the other groups. The amount of mature compact bone filling the defects was greater in the GT/M-5/B-3 and GT/M-6/B-3 groups than in the other groups. The present study demonstrated that the GT sponge loaded with MSCs and BMP-2 promoted bone regeneration in an equine bone defect model. The GT/MSC/BMP-2 described here may be useful for treating horses with bone injuries.

Production of Composite Scaffold Containing Silk Fibroin, Chitosan, and Gelatin for 3D Cell Culture and Bone Tissue Regeneration.

Science.gov (United States)

Li, Jianqing; Wang, Qiuke; Gu, Yebo; Zhu, Yu; Chen, Liang; Chen, Yunfeng

2017-11-08

BACKGROUND Bone tissue engineering, a powerful tool to treat bone defects, is highly dependent on use of scaffolds. Both silk fibroin (SF) and chitosan (Cs) are biocompatible and actively studied for reconstruction of tissue engineering. Gelatin (Gel) is also widely applied in the biomedical field due to its low antigenicity and physicochemical stability. MATERIAL AND METHODS In this study, 4 different types of scaffolds were constructed - SF, SF/Cs, SF/Gel, and SF/Cs/Gel - and we compared their physical and chemical properties as well as biological characterization of these scaffolds to determine the most suitable scaffold for use in bone regeneration. First, these scaffolds were produced via chemical cross-linking method and freeze-drying technique. Next, the characterization of internal structure was studied using scanning electron microscopy and the porosity was evaluated by liquid displacement method. Then, we compared physicochemical properties such as water absorption rate and degradation property. Finally, MC3T3-E1 cells were inoculated on the scaffolds to study the biocompatibility and osteogenesis of the three-dimensional (3D) scaffolds in vitro. RESULTS The composite scaffold formed by all 3 components was the best for use in bone regeneration. CONCLUSIONS We conclude that the best scaffold among the 4 studied for MC3T3-E1 cells is our SF/Cs/Gel scaffold, suggesting a new choice for bone regeneration that can be used to treat bone defects or fractures in clinical practice.

Calcium citrate: a new biomaterial that can enhance bone formation in situ

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WANG Li-ming

2012-11-01

Full Text Available 【Abstract】 Objective: To investigate the effect of a new biomaterial combining calcium citrate and recombinant human bone morphogenetic protein-2 (rhBMP-2 on bone regeneration in a bone defect rabbit model. Methods: Totally 30 male New Zealand white rabbits were randomly and equally divided into calcium citrate-rhBMP-2 (CC-rhBMP-2 group and rhBMP-2 only group. Two 10 mm-long and 5 mm-deep bone defects were respec-tively created in the left and right femoral condyles of the rabbits. Subsequently 5 pellets of calcium citrate (10 mg combined with rhBMP-2 (2 mg or rhBMP-2 alone were im-planted into the bone defects and compressed with cotton swab. Bone granules were obtained at 2, 4 and 6 weeks after procedure and received histological analysis. LSD t-test and a subsequent t-test were adopted for statistical analysis. Results: Histomorphometric analysis revealed newly formed bones, and calcium citrate has been absorbed in the treatment group. The percent of newly formed bone area in femoral condyle in control group and CC-rhBMP-2 group was respectively 31.73%±1.26% vs 48.21%±2.37% at 2 weeks; 43.40%±1.65% vs 57.32%±1.47% at 4 weeks, and 51.32%±7.80% vs 66.74%±4.05% at 6 weeks (P<0.05 for all. At 2 weeks, mature cancellous bone was observed to be already formed in the treatment group. Conclusion: From this study, it can be concluded that calcium citrate combined with rhBMP-2 signifcantly en-hances bone regeneration in bone defects. This synthetic gelatin matrix stimulates formation of new bone and bone marrow in the defect areas by releasing calcium ions. Key words: Bone morphogenetic protein-2; Biocompatible materials; Calcium citrate; Gelatin

Early matrix change of a nanostructured bone grafting substitute in the rat.

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Xu, Weiguo; Holzhüter, Gerd; Sorg, Heiko; Wolter, Daniel; Lenz, Solvig; Gerber, Thomas; Vollmar, Brigitte

2009-11-01

A nanocrystalline bone substitute embedded in a highly porous silica gel matrix (NanoBone) has previously been shown to bridge bone defects by an organic matrix. As the initial host response on the bone graft substitute might be a determinant for subsequent bone formation, our present purpose was to characterize the early tissue reaction on this biomaterial. After implantation of 80 mg of NanoBone into the adipose neck tissue of a total of 35 rats, grafts were harvested for subsequent analysis at days 3, 6, 9, 12, and 21. The biomaterial was found encapsulated by granulation tissue which partly penetrated the implant at day 3 and completely pervaded the graft at day 12 on implantation. Histology revealed tartrate-resistant acid phosphatase (TRAP)-positive giant cells covering the biomaterial. ED1 (CD68) immunopositivity of these cells further indicated their osteoclast-like phenotype. Scanning electron microscopy revealed organic tissue components within the periphery of the graft already at day 9, whereas the central hematoma region still presented the silica-surface of the biomaterial. Energy dispersive X-ray spectroscopy further demonstrated that the silica gel was degraded faster in the peripheral granulation tissue than in the central hematoma and was replaced by organic host components by day 12. In conclusion, the silica gel matrix is rapidly replaced by carbohydrate macromolecules. This might represent a key step in the process of graft degradation on its way toward induction of bone formation. The unique composition and structure of this nanoscaled biomaterial seem to support its degradation by host osteoclast-like giant cells.

Investigating Linear and Nonlinear Viscoelastic behaviour and microstructures of Gelatine-Multiwalled carbon nanotubes composites

KAUST Repository

Yang, Zhi

2015-12-01

We have investigated the linear and nonlinear rheology of various gelatine-multiwalled carbon nanotube (gel-MWNT) composites, namely physically-crosslinked-gelatine gel-MWNT composites, chemically-crosslinked-gelatine gel-MWNT composites, and chemically-physically-crosslinked-gelatine gel-MWNT composites. Further, the internal structures of these gel-MWNT composites were characterized by ultra-small angle neutron scattering and scanning electron microscopy. The adsorption of gelatine onto the surface of MWNT is also investigated to understand gelatine-assisted dispersion of MWNT during ultrasonication. For all gelatine gels, addition of MWNT increases their complex modulus. The dependence of storage modulus with frequency for gelatine-MWNT composites is similar to that of the corresponding neat gelatine matrix. However, by incorporating MWNT, the dependence of the loss modulus on frequency is reduced. The linear viscoelastic region is decreased approximately linearly with the increase of MWNT concentration. The pre-stress results demonstrate that the addition of MWNT does not change the strain-hardening behaviour of physically-crosslinked gelatine gel. However, the addition of MWNT can increase the strain-hardening behaviour of chemically-crosslinked gelatine gel, and chemically-physically crosslinked gelatine gel. Results from light microscopy, cryo-SEM, and USANS demonstrate the hierarchical structures of MWNT, including that tens-of-micron scale MWNT agglomerates are present. Furthermore, the adsorption curve of gelatine onto the surface of MWNT follows two-stage pseudo-saturation behaviour.

Matrix change of bone grafting substitute after implantation into guinea pig bulla.

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Punke, Ch; Zehlicke, T; Just, T; Holzhüter, G; Gerber, T; Pau, H W

2012-05-01

Many different surgical techniques have been developed to remove open mastoid cavities. In addition to autologous materials, alloplastic substances have been used. A very slow absorption of these materials and extrusion reactions have been reported. We investigated a newly developed, highly porous bone grafting material to eliminate open mastoid cavities, in an animal model. To characterise the transformation process, the early tissue reactions were studied in relation to the matrix transformation of the bone material. NanoBone (NB), a highly porous bone grafting material based on calcium phosphate and silica, was filled into the open bullae from 20 guinea pigs. The bullae were examined histologically. Energy dispersive X-ray spectroscopy (EDX) was used to investigate the change in the elemental composition at different sampling times. The surface topography of the sections was examined by electron microscopy. After 1 week, periodic acid-Schiffs (PAS) staining demonstrated accumulation of glycogen and proteins, particularly in the border area of the NB particles. After 2 weeks, the particles were evenly coloured after PAS staining. EDX analysis showed a rapid absorption of the silica in the bone grafting material. NanoBone showed a rapid matrix change after implantation in the bullae of guinea pigs. The absorption of the silica matrix and replacement by PAS-positive substances like glycoproteins and mucopolysaccharides seems to play a decisive role in the degradation processes of NB. This is associated with the good osteoinductive properties of the material.

Gelatin Scaffolds with Controlled Pore Structure and Mechanical Property for Cartilage Tissue Engineering.

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Chen, Shangwu; Zhang, Qin; Nakamoto, Tomoko; Kawazoe, Naoki; Chen, Guoping

2016-03-01

Engineering of cartilage tissue in vitro using porous scaffolds and chondrocytes provides a promising approach for cartilage repair. However, nonuniform cell distribution and heterogeneous tissue formation together with weak mechanical property of in vitro engineered cartilage limit their clinical application. In this study, gelatin porous scaffolds with homogeneous and open pores were prepared using ice particulates and freeze-drying. The scaffolds were used to culture bovine articular chondrocytes to engineer cartilage tissue in vitro. The pore structure and mechanical property of gelatin scaffolds could be well controlled by using different ratios of ice particulates to gelatin solution and different concentrations of gelatin. Gelatin scaffolds prepared from ≥70% ice particulates enabled homogeneous seeding of bovine articular chondrocytes throughout the scaffolds and formation of homogeneous cartilage extracellular matrix. While soft scaffolds underwent cellular contraction, stiff scaffolds resisted cellular contraction and had significantly higher cell proliferation and synthesis of sulfated glycosaminoglycan. Compared with the gelatin scaffolds prepared without ice particulates, the gelatin scaffolds prepared with ice particulates facilitated formation of homogeneous cartilage tissue with significantly higher compressive modulus. The gelatin scaffolds with highly open pore structure and good mechanical property can be used to improve in vitro tissue-engineered cartilage.

Radiation synthesis and characterization of nanosilver/gelatin/carboxymethyl chitosan hydrogel

International Nuclear Information System (INIS)

Zhou Ying; Zhao Yinghui; Wang Lu; Xu Ling; Zhai Maolin; Wei Shicheng

2012-01-01

A series of antibacterial hydrogels were fabricated from an aqueous solution of AgNO 3 , gelatin and carboxymethyl chitosan (CM-chitosan) by radiation-induced reduction and crosslinking at ambient temperature. The nanosilver particles were in situ synthesized accompanying with the formation of gelatin/CM-chitosan hydrogel. Transmission Electron Microscope and UV–vis analysis have verified the formation and homogeneous distribution of nanosilver particles in the hydrogel matrix. The nanosilver/gelatin/CM-chitosan hydrogels possessed interconnected porous structure, had a compressive modulus of 44 to 56 kPa, and could absorb 62 to 108 times of deionized water to its dry weight. Furthermore, the hydrogels were found to have sound antibacterial effect on Escherichia coli (E. coli), and their antibacterial ability could be significantly enhanced by the increasing of AgNO 3 content. The comprehensive results of this study suggest that nanosilver/gelatin/CM-chitosan hydrogels have potential as an antibacterial wound dressing. - Highlights: ► Nanosilver/gelatin/CM-chitosan hydrogel was synthesized by radiation crosslinking. ► Nanosilver particles distributed homogeneously in the hydrogel. ► The size of nanosilver increased with the increase of AgNO 3 concentration. ► The nanosilver/gelatin/CM-chitosan hydrogel has antibacterial ability.

Particle size modeling and morphology study of chitosan/gelatin/nanohydroxyapatite nanocomposite microspheres for bone tissue engineering.

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Bagheri-Khoulenjani, Shadab; Mirzadeh, Hamid; Etrati-Khosroshahi, Mohammad; Shokrgozar, Mohammad Ali

2013-06-01

In this study, nanocomposite microspheres based on chitosan/gelatin/nanohydroxyapatite were fabricated, and effects of the nanohydroxyapatite/biopolymer (chitosan/gelatin) weight ratio (nHA/P), stirring rate, chitosan concentration and biopolymer concentration on the particle size, and morphology of nanocomposite microspheres were investigated. Particle size of microspheres was modeled by design of experiments using the surface response method. Particle size, morphology of microspheres, and distribution of nanoparticles within the composite microspheres were evaluated using an optical microscope, scanning electron microscopy (SEM) and transmission electron microscopy (TEM), respectively. X-ray diffraction and Fourier transform infrared spectroscopy were applied to study the physical and chemical characteristics of microspheres. Results showed that by modulating the nHA/P ratio, chitosan concentration, polymer concentration, and stirring rate, it is possible to fabricate microspheres in wide rages of particle size (5-150 μm). Analysis of variance confirmed that the modified quadratic model can be used to predict the particle size of nanocomposite microspheres within the design space. SEM studies showed that microspheres with different compositions had totally different morphologies from dense morphologies to porous ones. TEM images demonstrated that nanoparticles were distributed uniformly within the polymeric matrix. MTT assay and cell culture studies showed that microspheres with different compositions possessed good biocompatibility. © 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2013. Copyright © 2012 Wiley Periodicals, Inc.

Investigating Effects of Gelatin-Chitosan Film on Culture of Bone Marrow Stromal Cells in Rat

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A Karami joyani

2015-02-01

Conclusion: Results of proliferation,differentiation and apoptosis cultured BMSCs on a gelatin-chitosan film showed that gelatin-chitosan film can be used as a good model of a biodegradable scaffold in tissue engineering and cell therapy.

Minimal invasive surgery for unicameral bone cyst using demineralized bone matrix: a case series

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Cho Hwan

2012-07-01

Full Text Available Abstract Background Various treatments for unicameral bone cyst have been proposed. Recent concern focuses on the effectiveness of closed methods. This study evaluated the effectiveness of demineralized bone matrix as a graft material after intramedullary decompression for the treatment of unicameral bone cysts. Methods Between October 2008 and June 2010, twenty-five patients with a unicameral bone cyst were treated with intramedullary decompression followed by grafting of demineralized bone matrix. There were 21 males and 4 female patients with mean age of 11.1 years (range, 3–19 years. The proximal metaphysis of the humerus was affected in 12 patients, the proximal femur in five, the calcaneum in three, the distal femur in two, the tibia in two, and the radius in one. There were 17 active cysts and 8 latent cysts. Radiologic change was evaluated according to a modified Neer classification. Time to healing was defined as the period required achieving cortical thickening on the anteroposterior and lateral plain radiographs, as well as consolidation of the cyst. The patients were followed up for mean period of 23.9 months (range, 15–36 months. Results Nineteen of 25 cysts had completely consolidated after a single procedure. The mean time to healing was 6.6 months (range, 3–12 months. Four had incomplete healing radiographically but had no clinical symptom with enough cortical thickness to prevent fracture. None of these four cysts needed a second intervention until the last follow-up. Two of 25 patients required a second intervention because of cyst recurrence. All of the two had a radiographical healing of cyst after mean of 10 additional months of follow-up. Conclusions A minimal invasive technique including the injection of DBM could serve as an excellent treatment method for unicameral bone cysts.

Minimal invasive surgery for unicameral bone cyst using demineralized bone matrix: a case series.

Science.gov (United States)

Cho, Hwan Seong; Seo, Sung Hwa; Park, So Hyun; Park, Jong Hoon; Shin, Duk Seop; Park, Il Hyung

2012-07-29

Various treatments for unicameral bone cyst have been proposed. Recent concern focuses on the effectiveness of closed methods. This study evaluated the effectiveness of demineralized bone matrix as a graft material after intramedullary decompression for the treatment of unicameral bone cysts. Between October 2008 and June 2010, twenty-five patients with a unicameral bone cyst were treated with intramedullary decompression followed by grafting of demineralized bone matrix. There were 21 males and 4 female patients with mean age of 11.1  years (range, 3-19 years). The proximal metaphysis of the humerus was affected in 12 patients, the proximal femur in five, the calcaneum in three, the distal femur in two, the tibia in two, and the radius in one. There were 17 active cysts and 8 latent cysts. Radiologic change was evaluated according to a modified Neer classification. Time to healing was defined as the period required achieving cortical thickening on the anteroposterior and lateral plain radiographs, as well as consolidation of the cyst. The patients were followed up for mean period of 23.9 months (range, 15-36 months). Nineteen of 25 cysts had completely consolidated after a single procedure. The mean time to healing was 6.6 months (range, 3-12 months). Four had incomplete healing radiographically but had no clinical symptom with enough cortical thickness to prevent fracture. None of these four cysts needed a second intervention until the last follow-up. Two of 25 patients required a second intervention because of cyst recurrence. All of the two had a radiographical healing of cyst after mean of 10 additional months of follow-up. A minimal invasive technique including the injection of DBM could serve as an excellent treatment method for unicameral bone cysts.

Marginal zinc deficiency in pregnant rats impairs bone matrix formation and bone mineralization in their neonates.

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Nagata, Masashi; Kayanoma, Megumu; Takahashi, Takeshi; Kaneko, Tetsuo; Hara, Hiroshi

2011-08-01

Zinc (Zn) deficiency during pregnancy may result in a variety of defects in the offspring. We evaluated the influence of marginal Zn deficiency during pregnancy on neonatal bone status. Nine-week-old male Sprague-Dawley rats were divided into two groups and fed AIN-93G-based experimental diets containing 35 mg Zn/kg (Zn adequately supplied, N) or 7 mg Zn/kg (low level of Zn, L) from 14-day preconception to 20 days of gestation, that is, 1 day before normal delivery. Neonates were delivered by cesarean section. Litter size and neonate weight were not different between the two groups. However, in the L-diet-fed dam group, bone matrix formation in isolated neonatal calvaria culture was clearly impaired and was not recovered by the addition of Zn into the culture media. Additionally, serum concentration of osteocalcin, as a bone formation parameter, was lower in neonates from the L-diet-fed dam group. Impaired bone mineralization was observed with a significantly lower content of phosphorus in neonate femurs from L-diet-fed dams compared with those from N-diet-fed dams. Moreover, Zn content in the femur and calvaria of neonates from the L-diet group was lower than that of the N-diet-fed group. In the marginally Zn-deficient dams, femoral Zn content, serum concentrations of Zn, and osteocalcin were reduced when compared with control dams. We conclude that maternal Zn deficiency causes impairment of bone matrix formation and bone mineralization in neonates, implying the importance of Zn intake during pregnancy for proper bone development of offspring.

Gelatinized wheat starch influences crystallization behaviour and structure of roll-in shortenings in laminated bakery products.

Science.gov (United States)

Mattice, Kristin D; Marangoni, Alejandro G

2018-03-15

One hydrogenated and one non-hydrogenated shortening were baked with isolated components of a croissant matrix, including crystalline wheat starch, gelatinized wheat starch, gluten, and formed gluten network. The impact of the matrix components on fat crystallization was analyzed for polymorphism using powder X-ray diffraction, solid fat content by pulsed nuclear magnetic resonance and thermal behaviour by differential scanning calorimetry. When compared to results obtained from croissants prepared with the respective shortenings, samples containing gelatinized wheat starch displayed notably similar results: polymorphic conversion, from the β' to β form over storage, and visually broader peaks in the melting endotherms indicating a greater temperature was required to completely melt all of the fat. All other component mixtures behaved similar to the respective fats in bulk. The measured rate of crystallization was greater in samples containing gelatinized wheat starch, indicating that the gelatinized starch could act as a nucleation site to speed crystallization. Copyright © 2017 Elsevier Ltd. All rights reserved.

Characterization of pre-gelatinized rice and bean flour

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Ana Vânia Carvalho

2013-06-01

Full Text Available The objective of this study was to develop a pre-gelatinized flour using a mixture of broken rice and split beans by thermoplastic extrusion, and to evaluate the physicochemical, nutritional, and technological quality of the final product. The extrusion parameters were maintained using three heating zones with temperatures of 30 ºC, 40 ºC, and 70 ºC; screw speed of 177 rpm; feed rate of 257 g/m, and circular matrix of 3.85 mm. The following characterization analyses were performed: physicochemical, nutritional, water absorption index (WAI, water solubility index (WSI, and paste viscosity. The pre-gelatinized rice and bean flour had an intermediate value of WAI, 7.51 g/g, and high WSI value, 24.61%. Regarding proteins, it was verified an average content of 12.9% in the final product. The amino acid contents found in the pre-gelatinized flour indicate that the mixture has the essential amino acids. It was also found that the pre-gelatinized flour supplies more than 60% of the essential amino acids recommended for children aged one to three years old. The gelatinized flour composed of broken rice and split beans is an alternative to the use of these by-products of the manufacture process of rice and beans to obtain a product with viable technological characteristics and high nutritional value.

Effects of fatigue on microstructure and mechanical properties of bone organic matrix under compression

International Nuclear Information System (INIS)

Trębacz, Hanna; Zdunek, Artur; Cybulska, Justyna; Pieczywek, Piotr

2013-01-01

The aim of the study was to investigate whether a fatigue induced weakening of cortical bone was revealed in microstructure and mechanical competence of demineralized bone matrix. Two types of cortical bone samples (plexiform and Haversian) were use. Bone slabs from the midshaft of bovine femora were subjected to cyclical bending. Fatigued and adjacent control samples were cut into cubes and demineralized in ethylenediaminetetraacetic acid. Demineralized samples were either subjected to microscopic quantitative image analysis, or compressed to failure (in longitudinal or transverse direction) with a simultaneous analysis of acoustic emission (AE). In fatigued samples porosity of organic matrix and average area of pores have risen, along with a change in the pores shape. The effect of fatigue depended on the type of the bone, being more pronounced in the plexiform than in Haversian tissue. Demineralized bone matrix was anisotropic under compressive loads in both types of cortical structure. The main result of fatigue pretreatment on mechanical parameters was a significant decrease of ultimate strain in the transverse direction in plexiform samples. The decrease of strain in this group was accompanied by a considerable increase of the fraction of large pores and a significant change in AE energy.

Assessment of gelatinases (MMP-2 and MMP-9) by gelatin zymography.

Science.gov (United States)

Toth, Marta; Sohail, Anjum; Fridman, Rafael

2012-01-01

Gelatin zymography is a simple yet powerful method to detect proteolytic enzymes capable of degrading gelatin from various biological sources. It is particularly useful for the assessment of two key members of the matrix metalloproteinase family, MMP-2 (gelatinase A) and MMP-9 (gelatinase B), due to their potent gelatin-degrading activity. This polyacrylamide gel electrophoresis-based method can provide a reliable assessment of the type of gelatinase, relative amount, and activation status (latent, compared with active enzyme forms) in cultured cells, tissues, and biological fluids. The method can be used to investigate factors that regulate gelatinase expression and modulate zymogen activation in experimental systems. The system provides information on the pattern of gelatinase expression and activation in human cancer tissues and how this relates to cancer progression. Interpretation of the data obtained in gelatin zymography requires a thorough understanding of the principles and pitfalls of the technique; this is particularly important when evaluating enzyme levels and the presence of active gelatinase species. If properly used, gelatin zymography is an excellent tool for the study of gelatinases in biological systems.

Radiation as a processing aid for upgrading gelatin and gelatin capsules

International Nuclear Information System (INIS)

Patel, K.M.; Tantry, M.; Sharma, G.; Gopal, N.G.S.

1979-01-01

Gelatin for pharmaceutical use should not contain more than 1000 microorganisms per gram and must not contain pathogenic microorganisms like Salmonella species and E.Coli. For some biological studies spore free gelatin is essential. Locally available gelatin and gelatin capsules have been found to contain a good number of microorganisms. Gelatin and its capsules containing different colours were exposed to graded doses of radiation and stored at different temperatures. They were then examined for microbial content, disintegration characteristics, pH, viscosity and colour change. The study shows that radiation treatment is useful to upgrade microbiologically both gelatin and its capsules. This technique provides apparently a useful and economic method for producing spore free gelatin which in commerce is relatively expensive. (auth.)

Radiation as a processing aid for upgrading gelatin and gelatin capsules

Energy Technology Data Exchange (ETDEWEB)

Patel, K M; Tantry, M; Sharma, G; Gopal, N G.S. [Bhabha Atomic Research Centre, Bombay (India). ISOMED

1979-01-01

Gelatin for pharmaceutical use should not contain more than 1000 microorganisms per gram and must not contain pathogenic microorganisms like Salmonella species and E.Coli. For some biological studies spore free gelatin is essential. Locally available gelatin and gelatin capsules have been found to contain a good number of microorganisms. Gelatin and its capsules containing different colours were exposed to graded doses of radiation and stored at different temperatures. They were then examined for microbial content, disintegration characteristics, pH, viscosity and colour change. The study shows that radiation treatment is useful to upgrade microbiologically both gelatin and its capsules. This technique provides apparently a useful and economic method for producing spore free gelatin which in commerce is relatively expensive.

The effect of carrier type on bone regeneration of demineralized bone matrix in vivo.

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Tavakol, Shima; Khoshzaban, Ahad; Azami, Mahmoud; Kashani, Iraj Ragerdi; Tavakol, Hani; Yazdanifar, Mahbube; Sorkhabadi, Seyed Mahdi Rezayat

2013-11-01

Demineralized bone matrix (DBM) is a bone substitute biomaterial used as an excellent grafting material. Some factors such as carrier type might affect the healing potential of this material. The background data discuss the present status of the field: Albumin as a main protein in blood and carboxymethyl cellulose (CMC) were applied frequently in the DBM gels. We investigated the bone-repairing properties of 2 DBMs with different carriers. Bone regeneration in 3 groups of rat calvaria treated with DBM from the Iranian Tissue Bank Research and Preparation Center, DBM from Hans Biomed Corporation, and an empty cavity was studied. Albumin and CMC as carriers were used. The results of bone regeneration in the samples after 1, 4, and 8 weeks of implantation were compared. The block of the histologic samples was stained with hematoxylin and eosin, and the percentage area of bone formation was calculated using the histomorphometry method. The results of in vivo tests showed a significantly stronger new regenerated bone occupation in the DBM with albumin carrier compared with the one with CMC 8 weeks after the implantation. The 2 types of DBM had a significant difference in bone regeneration. This difference is attributed to the type of carriers. Albumin could improve mineralization and bioactivity compared with CMC.

Bone regeneration by gelatin hydro-gel seat containing BMP-2 and its application to canine orbital floor fracture Model

International Nuclear Information System (INIS)

Mochizuki, Yuichi

2007-01-01

Reported are preparation of the gel seat in the title (GHG) for sustained release of BMP-2 (bone morphogenetic protein-2) and its application to the fracture model of orbital floor of the dog. The chemically linked GHG was prepared from gelatin and glutaraldehyde and lyophilized. BMP-2 solution was dropped on the GHG seat to be contained there. To see the biobehavior of BMP-2 and GHG, they were labeled with 125 I and were given subcutaneously in the back of nude mice, of which remaining radioactivity was periodically measured by Aloka ARC-310B gamma counter. This experiment revealed the sustained release of BMP-2 along with degradation of the GHG. Then a complex of the GHG and bio-degradable polymer (L-lactide-ε-caprolactone) was prepared and implanted to the artificially fractured region (10 x 10 mm) of dog orbit floor, of which recovering process was evaluated by analysis of bone structure with soft X-ray (SOFRON, TRS-1005) roentgenography, histology, and micro-CT imaging (Comscantecno's Scan Xmate-A090S) for trabecular bone volume, thickness, number and separation. This experiment revealed that new bone was effectively induced to regenerate on the complex, of which structure was found similar to the normal trabecula. Thus in future, the complex can be useful for ideal treatment of the orbit floor fracture without necessity of donor. (R.T.)

Amino Acid Profile, Group of Functional and Molecular Weight Distribution of Goat Skin Gelatin That Produced Through Acid Process

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Muhammad Irfan Said

2012-02-01

Full Text Available Gelatin is a product of hydrolysis of collagen protein from animals that are partially processed.  Gelatin used in food and non food industries.  Gelatin is produced when many import of raw skins and bones of pigs and cows.  Goat skins potential as a raw material substitution that still doubt its halal. Process production of gelatin determine the properties of gelatin. The objectives of this research were to determine amino acid profile, group of functional and molecular weight distribution of gelatin made from goat skins which was produced through a process of acid. The skin of male Bligon goat, 1.5 to 2.5 year old was used as raw materials. Process production of gelatin was using acid type acetic acid (CH3COOH 0.5 M (v/v as curing material. The experimental design applied in this study and commercial gelatin was used as control. The results showed that gelatin produced from goat skin through the process of acid had properties identical with commercial gelatin. It can be concluded that the gelatin has the potential substitute product of commercial gelatin. Keywords: collagen, gelatin, goat skin, curing, acid process

Heterogeneous and self-organizing mineralization of bone matrix promoted by hydroxyapatite nanoparticles.

Science.gov (United States)

Campi, G; Cristofaro, F; Pani, G; Fratini, M; Pascucci, B; Corsetto, P A; Weinhausen, B; Cedola, A; Rizzo, A M; Visai, L; Rea, G

2017-11-16

The mineralization process is crucial to the load-bearing characteristics of the bone extracellular matrix. In this work, we have studied the spatiotemporal dynamics of mineral deposition by human bone marrow mesenchymal stem cells differentiating toward osteoblasts promoted by the presence of exogenous hydroxyapatite nanoparticles. At the molecular level, the added nanoparticles positively modulated the expression of bone-specific markers and enhanced calcified matrix deposition during osteogenic differentiation. The nucleation, growth and spatial arrangement of newly deposited hydroxyapatite nanocrystals have been evaluated using scanning micro X-ray diffraction and scanning micro X-ray fluorescence. As leading results, we have found the emergence of a complex scenario where the spatial organization and temporal evolution of the process exhibit heterogeneous and self-organizing dynamics. At the same time the possibility of controlling the differentiation kinetics, through the addition of synthetic nanoparticles, paves the way to empower the generation of more structured bone scaffolds in tissue engineering and to design new drugs in regenerative medicine.

Hydrazine-induced thermo-reversible optical shifts in silver-gelatin bionanocomposites

Science.gov (United States)

Aimé, Carole; Rietveld, Ivo B.; Coradin, Thibaud

2011-03-01

Bionanocomposites formed by in situ growth of silver nanoparticles within gelatin gels exhibit large (up to 100 nm) thermo-reversible optical shifts resulting from the enhancement of gel matrix scattering by the interaction of the biopolymer with the hydrazine reducing agent.

Nonvolatile Resistive Switching Memory Utilizing Cobalt Embedded in Gelatin

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Cheng-Jung Lee

2017-12-01

Full Text Available This study investigates the preparation and electrical properties of Al/cobalt-embedded gelatin (CoG/ indium tin oxide (ITO resistive switching memories. Co. elements can be uniformly distributed in gelatin without a conventional dispersion procedure, as confirmed through energy dispersive X-ray analyzer and X-ray photoelectron spectroscopy observations. With an appropriate Co. concentration, Co. ions can assist the formation of an interfacial AlOx layer and improve the memory properties. High ON/OFF ratio, good retention capability, and good endurance switching cycles are demonstrated with 1 M Co. concentration, in contrast to 0.5 M and 2 M memory devices. This result can be attributed to the suitable thickness of the interfacial AlOx layer, which acts as an oxygen reservoir and stores and releases oxygen during switching. The Co. element in a solution-processed gelatin matrix has high potential for bio-electronic applications.

A 3D printed nano bone matrix for characterization of breast cancer cell and osteoblast interactions

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Zhu, Wei; Castro, Nathan J.; Cui, Haitao; Zhou, Xuan; Boualam, Benchaa; McGrane, Robert; Glazer, Robert I.; Zhang, Lijie Grace

2016-08-01

Bone metastasis is one of the most prevalent complications of late-stage breast cancer, in which the native bone matrix components, including osteoblasts, are intimately involved in tumor progression. The development of a successful in vitro model would greatly facilitate understanding the underlying mechanism of breast cancer bone invasion as well as provide a tool for effective discovery of novel therapeutic strategies. In the current study, we fabricated a series of in vitro bone matrices composed of a polyethylene glycol hydrogel and nanocrystalline hydroxyapatite of varying concentrations to mimic the native bone microenvironment for the investigation of breast cancer bone metastasis. A stereolithography-based three-dimensional (3D) printer was used to fabricate the bone matrices with precisely controlled architecture. The interaction between breast cancer cells and osteoblasts was investigated in the optimized bone matrix. Using a Transwell® system to separate the two cell lines, breast cancer cells inhibited osteoblast proliferation, while osteoblasts stimulated breast cancer cell growth, whereas, both cell lines increased IL-8 secretion. Breast cancer cells co-cultured with osteoblasts within the 3D bone matrix formed multi-cellular spheroids in comparison to two-dimensional monolayers. These findings validate the use of our 3D printed bone matrices as an in vitro metastasis model, and highlights their potential for investigating breast cancer bone metastasis.

Co-electrospun gelatin-poly(L-lactic acid) scaffolds: Modulation of mechanical properties and chondrocyte response as a function of composition

Energy Technology Data Exchange (ETDEWEB)

Torricelli, Paola [Preclinical and Surgical Studies Laboratory, Codivilla Putti Research Institute, Rizzoli Orthopaedic Institute, via di Barbiano, 1/10, 40136 Bologna (Italy); Laboratory of Biocompatibility, Innovative Technologies and Advanced Therapies—Department Rizzoli Research, Innovation, Technology, via di Barbiano, 1/10, 40136 Bologna (Italy); Gioffrè, Michela; Fiorani, Andrea; Panzavolta, Silvia [Department of Chemistry “G. Ciamician” and National Consortium of Materials Science and Technology (INSTM, Bologna RU), University of Bologna (Italy); Gualandi, Chiara [Department of Chemistry “G. Ciamician” and National Consortium of Materials Science and Technology (INSTM, Bologna RU), University of Bologna (Italy); Advanced Mechanics and Materials—Interdepartmental Center for Industrial Research (AMM ICIR), University of Bologna (Italy); Fini, Milena [Preclinical and Surgical Studies Laboratory, Codivilla Putti Research Institute, Rizzoli Orthopaedic Institute, via di Barbiano, 1/10, 40136 Bologna (Italy); Laboratory of Biocompatibility, Innovative Technologies and Advanced Therapies—Department Rizzoli Research, Innovation, Technology, via di Barbiano, 1/10, 40136 Bologna (Italy); Focarete, Maria Letizia, E-mail: marialetizia.focarete@unibo.it [Department of Chemistry “G. Ciamician” and National Consortium of Materials Science and Technology (INSTM, Bologna RU), University of Bologna (Italy); Health Sciences and Technologies—Interdepartmental Center for Industrial Research (HST-ICIR) (Italy); Bigi, Adriana [Department of Chemistry “G. Ciamician” and National Consortium of Materials Science and Technology (INSTM, Bologna RU), University of Bologna (Italy)

2014-03-01

Bio-synthetic scaffolds of interspersed poly(L-lactic acid) (PLLA) and gelatin (GEL) fibers are fabricated by co-electrospinning. Tailored PLLA/GEL compositions are obtained and GEL crosslinking with genipin provides for the maintenance of good fiber morphology. Scaffold tensile mechanical properties are intermediate between those of pure PLLA and GEL and vary as a function of PLLA content. Primary human chondrocytes grown on the scaffolds exhibit good proliferation and increased values of the differentiation parameters, especially for intermediate PLLA/GEL compositions. Mineralization tests enable the deposition of a uniform layer of poorly crystalline apatite onto the scaffolds, suggesting potential applications involving cartilage as well as cartilage–bone interface tissue engineering. - Highlights: • Bio-synthetic scaffolds of PLLA and gelatin are produced by co-electrospinning. • Scaffolds with tailored PLLA–gelatin composition are fabricated. • PLLA/gelatin ratio controls scaffold mechanical properties and mineralization. • Chondrocyte proliferation and differentiation are modulated. • Scaffolds are suitable for cartilage–bone interface tissue engineering.

Hybrid scaffold bearing polymer-siloxane Schiff base linkage for bone tissue engineering

Energy Technology Data Exchange (ETDEWEB)

Nair, Bindu P., E-mail: bindumelekkuttu@gmail.com; Gangadharan, Dhanya; Mohan, Neethu; Sumathi, Babitha; Nair, Prabha D., E-mail: pdnair49@gmail.com

2015-07-01

Scaffolds that can provide the requisite biological cues for the fast regeneration of bone are highly relevant to the advances in tissue engineering and regenerative medicine. In the present article, we report the fabrication of a chitosan–gelatin–siloxane scaffold bearing interpolymer-siloxane Schiff base linkage, through a single-step dialdehyde cross-linking and freeze-drying method using 3-aminopropyltriethoxysilane as the siloxane precursor. Swelling of the scaffolds in phosphate buffered saline indicates enhancement with increase in siloxane concentration, whereas compressive moduli of the wet scaffolds reveal inverse dependence, owing to the presence of siloxane, rich in silanol groups. It is suggested that through the strategy of dialdehyde cross-linking, a limiting siloxane loading of 20 wt.% into a chitosan-gelatin matrix should be considered ideal for bone tissue engineering, because the scaffold made with 30 wt.% siloxane loading degrades by 48 wt.%, in 21 days. The hybrid scaffolds bearing Schiff base linkage between the polymer and siloxane, unlike the stable linkages in earlier reports, are expected to give a faster release of siloxanes and enhancement in osteogenesis. This is verified by the in vitro evaluation of the hybrid scaffolds using rabbit adipose mesenchymal stem cells, which revealed osteogenic cell-clusters on a polymer-siloxane scaffold, enhanced alkaline phosphatase activity and the expression of bone-specific genes, whereas the control scaffold without siloxane supported more of cell-proliferation than differentiation. A siloxane concentration dependent enhancement in osteogenic differentiation is also observed. - Highlights: • A hybrid scaffold bearing interpolymer-siloxane Schiff base linkage • A limiting siloxane loading of 20 wt.% into chitosan–gelatin matrix • A siloxane concentration dependent enhancement in osteogenic differentiation.

Injection of demineralized bone matrix with bone marrow concentrate improves healing in unicameral bone cyst.

Science.gov (United States)

Di Bella, Claudia; Dozza, Barbara; Frisoni, Tommaso; Cevolani, Luca; Donati, Davide

2010-11-01

Unicameral bone cysts are benign lesions that usually spontaneously regress with skeletal maturity; however, the high risk of pathologic fractures often justifies treatment that could reinforce a weakened bone cortex. Various treatments have been proposed but there is no consensus regarding the best procedure. We compared the healing rates and failures of two methods of cure based on multiple injections of corticosteroid or a single injection of demineralized bone matrix (DBM) in association with bone marrow concentrate (BMC). We retrospectively reviewed 184 patients who had one of the two treatments for unicameral bone cysts with cortical erosion. Clinical records were reviewed for treatment failures and radiographs for healing in all patients. The minimum followup was 12 months for the Steroids Group (mean, 48 months; range, 12-120 months) and 12 months for the DBM + BMC Group (mean, 20 months; range, 12-28 months). After one treatment we observed a lower healing rate of cysts treated with multiple injections of steroids compared with the healing after the first injection of DBM + BMC (21% versus 58%, respectively). At last followup, 38% healed with steroids and 71% with DBM + BMC. The rate of failure after one steroid injection was higher than after a single injection of BDM + BMC (63% versus 24%, respectively). We observed no difference in fracture rates after treatment between the two groups. A single injection of DBM added with autologous bone marrow concentrate appears to provide a higher healing rate with a lower number of failures compared with a single injection of steroids.

Novel electrospun nanofibers of modified gelatin-tyrosine in cartilage tissue engineering

International Nuclear Information System (INIS)

Agheb, Maria; Dinari, Mohammad; Rafienia, Mohammad; Salehi, Hossein

2017-01-01

In natural cartilage tissues, chondrocytes are linked to extracellular matrix (ECM) through cell-surface binding proteins. Surface modification of gelatin can provide a new generation of biopolymers and fibrous scaffolds with chemical, mechanical, and biological properties. In this study tyrosine protein and 1,2,3-triazole ring were utilized to functionalize gelatin without Cu catalyst. Their molecular structure was characterized by Fourier transform infrared spectroscopy (FTIR) and nuclear magnetic resonance spectroscopy ( 1 HNMR). Chemical cross-linkers such as glutaraldehyde (GA) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC)/N-hydroxysulfosuccinimide (NHS) were used to electrospin the modified gelatin. The modification of gelatin and cross-linking effects were confirmed by scanning electron microscopy (SEM), contact angle measurement, and mechanical tests. MTT assay using chondrocyte cells showed cell viability of electrospun modified gelatin scaffolds. In vitro cell culture studies showed that electrospun engineered protein scaffolds would support the attachment and growth of cells. The results also showed that cross-linked nanofibers with EDC/NHS could be considered excellent matrices in cell adhesion and proliferation before electrospinning process and their potential substrate in tissue engineering applications, especially in the field of cartilage engineering.

Novel electrospun nanofibers of modified gelatin-tyrosine in cartilage tissue engineering

Energy Technology Data Exchange (ETDEWEB)

Agheb, Maria [Biosensor Research Center, Isfahan University of Medical Sciences, Isfahan 81744176 (Iran, Islamic Republic of); Dinari, Mohammad [Department of Chemistry, Isfahan University of Technology, Isfahan 84156-83111 (Iran, Islamic Republic of); Rafienia, Mohammad, E-mail: m_rafienia@med.mui.ac.ir [Biosensor Research Center, Isfahan University of Medical Sciences, Isfahan 81744176 (Iran, Islamic Republic of); Salehi, Hossein [Biosensor Research Center, Isfahan University of Medical Sciences, Isfahan 81744176 (Iran, Islamic Republic of)

2017-02-01

In natural cartilage tissues, chondrocytes are linked to extracellular matrix (ECM) through cell-surface binding proteins. Surface modification of gelatin can provide a new generation of biopolymers and fibrous scaffolds with chemical, mechanical, and biological properties. In this study tyrosine protein and 1,2,3-triazole ring were utilized to functionalize gelatin without Cu catalyst. Their molecular structure was characterized by Fourier transform infrared spectroscopy (FTIR) and nuclear magnetic resonance spectroscopy ({sup 1}HNMR). Chemical cross-linkers such as glutaraldehyde (GA) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC)/N-hydroxysulfosuccinimide (NHS) were used to electrospin the modified gelatin. The modification of gelatin and cross-linking effects were confirmed by scanning electron microscopy (SEM), contact angle measurement, and mechanical tests. MTT assay using chondrocyte cells showed cell viability of electrospun modified gelatin scaffolds. In vitro cell culture studies showed that electrospun engineered protein scaffolds would support the attachment and growth of cells. The results also showed that cross-linked nanofibers with EDC/NHS could be considered excellent matrices in cell adhesion and proliferation before electrospinning process and their potential substrate in tissue engineering applications, especially in the field of cartilage engineering.

Patient-Derived Human Induced Pluripotent Stem Cells From Gingival Fibroblasts Composited With Defined Nanohydroxyapatite/Chitosan/Gelatin Porous Scaffolds as Potential Bone Graft Substitutes.

Science.gov (United States)

Ji, Jun; Tong, Xin; Huang, Xiaofeng; Zhang, Junfeng; Qin, Haiyan; Hu, Qingang

2016-01-01

Human embryonic stem cells and adult stem cells have always been the cell source for bone tissue engineering. However, their limitations are obvious, including ethical concerns and/or a short lifespan. The use of human induced pluripotent stem cells (hiPSCs) could avoid these problems. Nanohydroxyapatite (nHA) is an important component of natural bone and bone tissue engineering scaffolds. However, its regulation on osteogenic differentiation with hiPSCs from human gingival fibroblasts (hGFs) is unknown. The purpose of the present study was to investigate the osteogenic differentiation of hiPSCs from patient-derived hGFs regulated by nHA/chitosan/gelatin (HCG) scaffolds with different nHA ratios, such as HCG-111 (1 wt/vol% nHA) and HCG-311 (3 wt/vol% nHA). First, hGFs were reprogrammed into hiPSCs, which have enhanced osteogenic differentiation capability. Second, HCG-111 and HCG-311 scaffolds were successfully synthesized. Finally, hiPSC/HCG complexes were cultured in vitro or subcutaneously transplanted into immunocompromised mice in vivo. The osteogenic differentiation effects of two types of HCG scaffolds on hiPSCs were assessed for up to 12 weeks. The results showed that HCG-311 increased osteogenic-related gene expression of hiPSCs in vitro proved by quantitative real-time polymerase chain reaction, and hiPSC/HCG-311 complexes formed much bone-like tissue in vivo, indicated by cone-beam computed tomography imaging, H&E staining, Masson staining, and RUNX-2, OCN immunohistochemistry staining. In conclusion, our study has shown that osteogenic differentiation of hiPSCs from hGFs was improved by HCG-311. The mechanism might be that the nHA addition stimulates osteogenic marker expression of hiPSCs from hGFs. Our work has provided an innovative autologous cell-based bone tissue engineering approach with soft tissues such as clinically abundant gingiva. The present study focused on patient-personalized bone tissue engineering. Human induced pluripotent stem cells

Development of a gelatin-based polyurethane vascular graft by spray, phase-inversion technology

International Nuclear Information System (INIS)

Losi, Paola; Al Kayal, Tamer; Briganti, Enrica; Volpi, Silvia; Soldani, Giorgio; Mancuso, Luisa; Cao, Giacomo; Celi, Simona; Gualerzi, Alice

2015-01-01

The capacity of a composite vascular graft constituting polyurethane (PU) and gelatin to support cell growth was investigated using human mesenchymal stem cells (hMSCs). Gelatin-based polyurethane grafts were fabricated by co-spraying polyurethane and gelatin using a spray, phase-inversion technique. Graft microstructure was investigated by light and scanning electron microscopy. Uniaxial tensile tests were performed to assess the grafts’ mechanical properties in longitudinal and circumferential directions. hMSCs obtained from bone marrow aspirate were seeded onto flat graft samples. After 24, 48, and 72 h of incubation, cell morphology was evaluated by Giemsa staining and cell viability was calculated by XTT assay. SEM analysis evidenced that PU samples display a microporous structure, whereas the gelatin-based PU samples show a fibrillar appearance. The presence of cross-linked gelatin produced a significant increase of ultimate tensile strength and ultimate elongation in circumferential directions compared to PU material. Qualitative analysis of hMSC adhesion onto the grafts revealed remarkable differences between gelatin-based PU and control graft. hMSCs grown onto gelatin-based PU graft form a monolayer that reached confluence at 72 h, whereas cells seeded onto the control graft were not able to undergo appropriate spreading. hMSCs grown onto gelatin-based PU graft showed significantly higher viability than cells seeded onto bare PU at all time points. In conclusion, a composite vascular graft was successfully manufactured by simultaneous co-spraying of a synthetic polymer and a protein to obtain a scaffold that combines the mechanical characteristics of polyurethanes with the favorable cell interaction features of gelatin. (paper)

Detection of growth factor binding to gelatin and heparin using a photonic crystal optical biosensor

International Nuclear Information System (INIS)

Morgan, Abby W.; Chan, Leo L.; Sendemir-Urkmez, Aylin; Cunningham, Brian T.; Jamison, Russell D.

2010-01-01

Drug-carrier interactions are important to protein controlled release systems to protect the protein from denaturation and ensure properly timed release. A novel photonic crystal biosensor was used to investigate a gelatin-protein controlled release system to determine the amount of protein bound to the carrier at physiological conditions. The Biomolecular Interaction Detection (BIND) system reflects a narrow band of wavelengths when white light is shone incident to the grating. As mass is deposited onto the surface, the peak wavelength value is shifted due to changes in the optical density of the biosensor. The BIND system was used to detect the binding of growth factors onto acidic gelatin, basic gelatin, and heparin on the sensor surface. Through a series of experiments, including functionalizing the sensor, adjusting the ionic strength of the solution, adjusting the substrate concentration, and minimizing non-specific signal, the adsorption of the gelatins and heparin on the sensor was enhanced. The binding interaction of recombinant human transforming growth factor (rhTGF)-β1 and bone morphogenetic protein (rhBMP)-2 with the two types of gelatin and heparin were investigated. The strength of the interaction between rhTGF-β1 and the substrates is in the following order: heparin > acidic gelatin > basic gelatin. RhBMP-2 bound to the substrates but with less intensity than TGF-β1: heparin > basic gelatin > acidic gelatin. This work provides support for the controlled release mechanism through degradation of the gelatin carrier.

Biological effect of hydrolyzed collagen on bone metabolism.

Science.gov (United States)

Daneault, Audrey; Prawitt, Janne; Fabien Soulé, Véronique; Coxam, Véronique; Wittrant, Yohann

2017-06-13

Osteoporosis is a chronic and asymptomatic disease characterized by low bone mass and skeletal microarchitectural deterioration, increased risk of fracture, and associated comorbidities most prevalent in the elderly. Due to an increasingly aging population, osteoporosis has become a major health issue requiring innovative disease management. Proteins are important for bone by providing building blocks and by exerting specific regulatory function. This is why adequate protein intake plays a considerable role in both bone development and bone maintenance. More specifically, since an increase in the overall metabolism of collagen can lead to severe dysfunctions and a more fragile bone matrix and because orally administered collagen can be digested in the gut, cross the intestinal barrier, enter the circulation, and become available for metabolic processes in the target tissues, one may speculate that a collagen-enriched diet provides benefits for the skeleton. Collagen-derived products such as gelatin or hydrolyzed collagen (HC) are well acknowledged for their safety from a nutritional point of view; however, what is their impact on bone biology? In this manuscript, we critically review the evidence from literature for an effect of HC on bone tissues in order to determine whether HC may represent a relevant alternative in the design of future nutritional approaches to manage osteoporosis prevention.

Autologous bone graft versus demineralized bone matrix in internal fixation of ununited long bones.

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Pieske, Oliver; Wittmann, Alexandra; Zaspel, Johannes; Löffler, Thomas; Rubenbauer, Bianka; Trentzsch, Heiko; Piltz, Stefan

2009-12-15

Non-unions are severe complications in orthopaedic trauma care and occur in 10% of all fractures. The golden standard for the treatment of ununited fractures includes open reduction and internal fixation (ORIF) as well as augmentation with autologous-bone-grafting. However, there is morbidity associated with the bone-graft donor site and some patients offer limited quantity or quality of autologous-bone graft material. Since allogene bone-grafts are introduced on the market, this comparative study aims to evaluate healing characteristics of ununited bones treated with ORIF combined with either iliac-crest-autologous-bone-grafting (ICABG) or demineralized-bone-matrix (DBM). From 2000 to 2006 out of sixty-two consecutive patients with non-unions presenting at our Level I Trauma Center, twenty patients had ununited diaphyseal fractures of long bones and were treated by ORIF combined either by ICABG- (n = 10) or DBM-augmentation (n = 10). At the time of index-operation, patients of the DBM-group had a higher level of comorbidity (ASA-value: p = 0.014). Mean duration of follow-up was 56.6 months (ICABG-group) and 41.2 months (DBM-group). All patients were clinically and radiographically assessed and adverse effects related to bone grafting were documented. The results showed that two non-unions augmented with ICABG failed osseous healing (20%) whereas all non-unions grafted by DBM showed successful consolidation during the first year after the index operation (p = 0.146). No early complications were documented in both groups but two patients of the ICABG-group suffered long-term problems at the donor site (20%) (p = 0.146). Pain intensity were comparable in both groups (p = 0.326). However, patients treated with DBM were more satisfied with the surgical procedure (p = 0.031). With the use of DBM, the costs for augmentation of the non-union-site are more expensive compared to ICABG (calculated difference: 160 euro/case). Nevertheless, this study demonstrated that the

Performance of high amylose starch-composited gelatin films influenced by gelatinization and concentration.

Science.gov (United States)

Wang, Wenhang; Wang, Kun; Xiao, Jingdong; Liu, Yaowei; Zhao, Yana; Liu, Anjun

2017-01-01

In order to study the impact of starch in film performance, high amylose corn starch was composited in gelatin films under different gelatinization conditions and, in high and low concentrations (10 and 50wt.%). It was found that hot water gelatinized starch (Gel-Shw) increased film mechanical strength and was dependent upon the starch concentration. The addition of an alkali component to the starch significantly enhanced the swelling of the starch granules and expedited the gelatinization process. Incorporation of starch, especially the alkalized starch (Sha), into the gelatin films decreased film solubility which improved its water resistance and water vapor permeability (WVP). Multiple techniques (DSC, TGA, FT-IR, and XRD) were used to characterize the process and results, including the crosslinking of the dissolved starch molecules and the particles formed from gelatinized starch during retrogradation process, which played an important role in improving the thermal stability of the composited gelatin films. Overall, the starch-gelatin composition provides a potential approach to improve gelatin film performance and benefit its applications in the food industry. Copyright © 2016 Elsevier B.V. All rights reserved.

Age-dependence of power spectral density and fractal dimension of bone mineralized matrix in atomic force microscope topography images: potential correlates of bone tissue age and bone fragility in female femoral neck trabeculae.

Science.gov (United States)

Milovanovic, Petar; Djuric, Marija; Rakocevic, Zlatko

2012-11-01

There is an increasing interest in bone nano-structure, the ultimate goal being to reveal the basis of age-related bone fragility. In this study, power spectral density (PSD) data and fractal dimensions of the mineralized bone matrix were extracted from atomic force microscope topography images of the femoral neck trabeculae. The aim was to evaluate age-dependent differences in the mineralized matrix of human bone and to consider whether these advanced nano-descriptors might be linked to decreased bone remodeling observed by some authors and age-related decline in bone mechanical competence. The investigated bone specimens belonged to a group of young adult women (n = 5, age: 20-40 years) and a group of elderly women (n = 5, age: 70-95 years) without bone diseases. PSD graphs showed the roughness density distribution in relation to spatial frequency. In all cases, there was a fairly linear decrease in magnitude of the power spectra with increasing spatial frequencies. The PSD slope was steeper in elderly individuals (-2.374 vs. -2.066), suggesting the dominance of larger surface morphological features. Fractal dimension of the mineralized bone matrix showed a significant negative trend with advanced age, declining from 2.467 in young individuals to 2.313 in the elderly (r = 0.65, P = 0.04). Higher fractal dimension in young women reflects domination of smaller mineral grains, which is compatible with the more freshly remodeled structure. In contrast, the surface patterns in elderly individuals were indicative of older tissue age. Lower roughness and reduced structural complexity (decreased fractal dimension) of the interfibrillar bone matrix in the elderly suggest a decline in bone toughness, which explains why aged bone is more brittle and prone to fractures. © 2012 The Authors Journal of Anatomy © 2012 Anatomical Society.

One-step direct-laser metal writing of sub-100 nm 3D silver nanostructures in a gelatin matrix

International Nuclear Information System (INIS)

Kang, SeungYeon; Vora, Kevin; Mazur, Eric

2015-01-01

Developing an ability to fabricate high-resolution, 3D metal nanostructures in a stretchable 3D matrix is a critical step to realizing novel optoelectronic devices such as tunable bulk metal-dielectric optical devices and THz metamaterial devices that are not feasible with alternative techniques. We report a new chemistry method to fabricate high-resolution, 3D silver nanostructures using a femtosecond-laser direct metal writing technique. Previously, only fabrication of 3D polymeric structures or single-/few-layer metal structures was possible. Our method takes advantage of unique gelatin properties to overcome such previous limitations as limited freedom in 3D material design and short sample lifetime. We fabricate more than 15 layers of 3D silver nanostructures with a resolution of less than 100 nm in a stable dielectric matrix that is flexible and has high large transparency that is well-matched for potential applications in the optical and THz metamaterial regimes. This is a single-step process that does not require any further processing. This work will be of interest to those interested in fabrication methods that utilize nonlinear light–matter interactions and the realization of future metamaterials. (fast track communication)

One-step direct-laser metal writing of sub-100 nm 3D silver nanostructures in a gelatin matrix

Science.gov (United States)

Kang, SeungYeon; Vora, Kevin; Mazur, Eric

2015-03-01

Developing an ability to fabricate high-resolution, 3D metal nanostructures in a stretchable 3D matrix is a critical step to realizing novel optoelectronic devices such as tunable bulk metal-dielectric optical devices and THz metamaterial devices that are not feasible with alternative techniques. We report a new chemistry method to fabricate high-resolution, 3D silver nanostructures using a femtosecond-laser direct metal writing technique. Previously, only fabrication of 3D polymeric structures or single-/few-layer metal structures was possible. Our method takes advantage of unique gelatin properties to overcome such previous limitations as limited freedom in 3D material design and short sample lifetime. We fabricate more than 15 layers of 3D silver nanostructures with a resolution of less than 100 nm in a stable dielectric matrix that is flexible and has high large transparency that is well-matched for potential applications in the optical and THz metamaterial regimes. This is a single-step process that does not require any further processing. This work will be of interest to those interested in fabrication methods that utilize nonlinear light-matter interactions and the realization of future metamaterials.

Biological conduits combining bone marrow mesenchymal stem cells and extracellular matrix to treat long-segment sciatic nerve defects

Directory of Open Access Journals (Sweden)

Yang Wang

2015-01-01

Full Text Available The transplantation of polylactic glycolic acid conduits combining bone marrow mesenchymal stem cells and extracellular matrix gel for the repair of sciatic nerve injury is effective in some respects, but few data comparing the biomechanical factors related to the sciatic nerve are available. In the present study, rabbit models of 10-mm sciatic nerve defects were prepared. The rabbit models were repaired with autologous nerve, a polylactic glycolic acid conduit + bone marrow mesenchymal stem cells, or a polylactic glycolic acid conduit + bone marrow mesenchymal stem cells + extracellular matrix gel. After 24 weeks, mechanical testing was performed to determine the stress relaxation and creep parameters. Following sciatic nerve injury, the magnitudes of the stress decrease and strain increase at 7,200 seconds were largest in the polylactic glycolic acid conduit + bone marrow mesenchymal stem cells + extracellular matrix gel group, followed by the polylactic glycolic acid conduit + bone marrow mesenchymal stem cells group, and then the autologous nerve group. Hematoxylin-eosin staining demonstrated that compared with the polylactic glycolic acid conduit + bone marrow mesenchymal stem cells group and the autologous nerve group, a more complete sciatic nerve regeneration was found, including good myelination, regularly arranged nerve fibers, and a completely degraded and resorbed conduit, in the polylactic glycolic acid conduit + bone marrow mesenchymal stem cells + extracellular matrix gel group. These results indicate that bridging 10-mm sciatic nerve defects with a polylactic glycolic acid conduit + bone marrow mesenchymal stem cells + extracellular matrix gel construct increases the stress relaxation under a constant strain, reducing anastomotic tension. Large elongations under a constant physiological load can limit the anastomotic opening and shift, which is beneficial for the regeneration and functional reconstruction of sciatic nerve. Better

Demineralized bone matrix and human cancellous bone enhance fixation of titanium implants

DEFF Research Database (Denmark)

Babiker, Hassan; Ding, Ming; Overgaard, Søren

Best Poster 5Demineralized bone matrix and human cancellous bone enhance fixation of titanium implants AuthorsBabiker , H.; Ding M.; Overgaard S.InstitutionOrthopaedic Research Laboratory, Department of Orthopaedic Surgery, Odense University Hospital, Clinical Institute, University of Southern...... from human tissue were included (IsoTis OrthoBiologics, Inc. USA). Both materials are commercially available. Titanium alloy implants (Biomet Inc.) of 10 mm in length and 10 mm in diameter were inserted bilaterally into the femoral condyles of 8 skeletally mature sheep. Thus four implants...... with a concentric gap of 2 mm were implanted in each sheep. The gap was filled with: DBM; DBM/CB with ratio of 1/3; DBM/allograft with ratio of 1/3; or allograft (Gold standard), respectively. Standardised surgical procedure was used1. At sacrifice, 6 weeks after surgery, both distal femurs were harvested...

Comparison of cell behavior on pva/pva-gelatin electrospun nanofibers with random and aligned configuration

Science.gov (United States)

Huang, Chen-Yu; Hu, Keng-Hsiang; Wei, Zung-Hang

2016-12-01

Electrospinning technique is able to create nanofibers with specific orientation. Poly(vinyl alcohol) (PVA) have good mechanical stability but poor cell adhesion property due to the low affinity of protein. In this paper, extracellular matrix, gelatin is incorporated into PVA solution to form electrospun PVA-gelatin nanofibers membrane. Both randomly oriented and aligned nanofibers are used to investigate the topography-induced behavior of fibroblasts. Surface morphology of the fibers is studied by optical microscopy and scanning electron microscopy (SEM) coupled with image analysis. Functional group composition in PVA or PVA-gelatin is investigated by Fourier Transform Infrared (FTIR). The morphological changes, surface coverage, viability and proliferation of fibroblasts influenced by PVA and PVA-gelatin nanofibers with randomly orientated or aligned configuration are systematically compared. Fibroblasts growing on PVA-gelatin fibers show significantly larger projected areas as compared with those cultivated on PVA fibers which p-value is smaller than 0.005. Cells on PVA-gelatin aligned fibers stretch out extensively and their intracellular stress fiber pull nucleus to deform. Results suggest that instead of the anisotropic topology within the scaffold trigger the preferential orientation of cells, the adhesion of cell membrane to gelatin have substantial influence on cellular behavior.

Demineralized bone matrix fibers formable as general and custom 3D printed mold-based implants for promoting bone regeneration.

Science.gov (United States)

Rodriguez, Rudy U; Kemper, Nathan; Breathwaite, Erick; Dutta, Sucharita M; Hsu, Erin L; Hsu, Wellington K; Francis, Michael P

2016-07-26

Bone repair frequently requires time-consuming implant construction, particularly when using un-formed implants with poor handling properties. We therefore developed osteoinductive, micro-fibrous surface patterned demineralized bone matrix (DBM) fibers for engineering both defect-matched and general three-dimensional implants. Implant molds were filled with demineralized human cortical bone fibers there were compressed and lyophilized, forming mechanically strong shaped DBM scaffolds. Enzyme linked immunosorbent assays and mass spectrometry confirmed that DBM fibers contained abundant osteogenic growth factors (bone morphogenetic proteins, insulin-like growth factor-I) and extracellular matrix proteins. Mercury porosimetry and mechanical testing showed interconnected pores within the mechanically stable, custom DBM fiber scaffolds. Mesenchymal stem cells readily attached to the DBM and showed increasing metabolic activity over time. DBM fibers further increased alkaline phosphatase activity in C2C12 cells. In vivo, DBM implants elicited osteoinductive potential in a mouse muscle pouch, and also promoted spine fusion in a rat arthrodesis model. DBM fibers can be engineered into custom-shaped, osteoinductive and osteoconductive implants with potential for repairing osseous defects with precise fitment, potentially reducing operating time. By providing pre-formed and custom implants, this regenerative allograft may improve patient outcomes following surgical bone repair, while further advancing personalized orthopedic and craniomaxillofacial medicine using three-dimensional-printed tissue molds.

Physicochemical and antifungal properties of bio-nanocomposite film based on gelatin-chitin nanoparticles.

Science.gov (United States)

Sahraee, Samar; Milani, Jafar M; Ghanbarzadeh, Babak; Hamishehkar, Hamed

2017-04-01

The gelatin-based nanocomposite films containing chitin nanoparticles (N-chitin) with concentrations of 0, 3, 5 and 10% were prepared and their physical, thermal and anti-microbial properties were investigated. Scanning electron microscopy (SEM) micrographs showed that N-chitin size distribution was around 60-70nm which dispersed appropriately at low concentration in gelatin matrix. The results showed that incorporation of N-chitin significantly influenced apparent color and transparency of the gelatin films. The reduced water vapor permeability (WVP) and solubility and higher surface hydrophobicity of the nanocomposite films were obtained by enhancing N-chitin concentration in film formulation. The use of N-chitin up to 5% concentration in the gelatin based nanocomposite film led to improved mechanical properties. Also, the results of differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) confirmed improved stability of nanocomposite films against melting and degradation at high temperatures in comparison to neat gelatin film. The well compatibility of chitin nanoparticles with gelatin polymer was concluded from Fourier transform infrared (FTIR) spectra and X-ray diffraction (XRD) plots. Finally, the gelatin based nanocomposite films had anti-fungal properties against Aspergillus niger in the contact surface zone. Increasing the concentration of N-chitin up to 5% enlarged inhibition zone diameter, but the nanocomposite film containing 10% N-chitin showed smaller inhibition zone. Copyright © 2016 Elsevier B.V. All rights reserved.

Healing improvement after rotator cuff repair using gelatin-grafted poly(L-lactide) electrospun fibrous membranes.

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Zhao, Song; Xie, Xiaoxing; Pan, Guoqing; Shen, Peng; Zhao, Jinzhong; Cui, Wenguo

2015-01-01

Rotator cuff tears (RCTs) are a common cause of shoulder pain and disability in middle and older age. Despite improvements in the understanding of this disease process and advances in surgical treatment, rotator cuff (RC) repair failure rates remain high. Insufficient healing capacity is likely the main factor for failure of reconstruction. We fabricated implantable biodegradable gelatin-grafted poly(L-lactide) (PLLA) fibrous membranes using electrospinning technology and evaluated them using in vitro cell proliferation assays. Then, we established chronic rat RCT models and randomly assigned rats into one of three groups. In group 1 (n = 48), the detached supraspinatus tendon was repaired to its anatomic footprint (transosseous repair). In groups 2 and 3, the rats underwent transosseous repair and were implanted with either pure PLLA membranes (n = 48) or gelatin-PLLA membranes (n = 48) to augment the repairs. The animals were killed at 2, 4, and 8 wk postoperatively, which was followed by histomorphometric and biomechanical evaluation. Histologic observations revealed that gelatin-PLLA membranes have excellent biocompatibility and biodegradability. At 2, 4, and 8 wk postoperatively, the gelatin-PLLA membranes significantly increased the area of glycosaminoglycan staining at the tendon-bone interface compared with the control group (P < 0.05) and significantly improved collagen organization, as measured by birefringence under polarized light at the healing enthesis compared with the control and PLLA groups (P < 0.05). Biomechanical testing revealed that the gelatin-PLLA group had a greater ultimate load to failure and stiffness than the control group at 4 and 8 wk (P < 0.05). The gelatin-PLLA membranes had the highest stress of the healing enthesis. Local application of gelatin-PLLA fibrous membranes to the healing tendon-bone interface after RC repair in a rat chronic RCT model was found to strengthen the healing enthesis, increase the area of

Nanoporous Structure of Bone Matrix at Osteoporosis from Data of Atomic Force Microscopy and IR Spectroscopy

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A. A. Gaidash

2011-01-01

Full Text Available It was found that in an osteoporotic bone the fraction of nanosized pores decreases, the mineral phase amorphizes, hydrated shells around mineralized particles of the bone matrix thicken, and adhesion forces increase. This contributes to the formation of water clusters similar to bulk water clusters compared to the healthy bone tissue and leads to the accumulation of more viscous liquid with increased intermolecular interaction forces in the pores of the bone matrix. Given this, the rates of chemical reactions proceeding in the water phase of ultrathin channels of general parts of collagen fibrils decrease. Ultimately, nanopores of collagen-apatite interfaces lose, to a certain extent, the capability of catalyzing the hydroxyapatite crystallization.

Changes in chemical composition of bone matrix in ovariectomized (OVX) rats detected by Raman spectroscopy and multivariate analysis

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Oshima, Yusuke; Iimura, Tadahiro; Saitou, Takashi; Imamura, Takeshi

2015-02-01

Osteoporosis is a major bone disease that connotes the risk of fragility fractures resulting from alterations to bone quantity and/or quality to mechanical competence. Bone strength arises from both bone quantity and quality. Assessment of bone quality and bone quantity is important for prediction of fracture risk. In spite of the two factors contribute to maintain the bone strength, only one factor, bone mineral density is used to determine the bone strength in the current diagnosis of osteoporosis. On the other hand, there is no practical method to measure chemical composition of bone tissue including hydroxyapatite and collagen non-invasively. Raman spectroscopy is a powerful technique to analyze chemical composition and material properties of bone matrix non-invasively. Here we demonstrated Raman spectroscopic analysis of the bone matrix in osteoporosis model rat. Ovariectomized (OVX) rat was made and the decalcified sections of tibias were analyzed by a Raman microscope. In the results, Raman bands of typical collagen appeared in the obtained spectra. Although the typical mineral bands at 960 cm-1 (Phosphate) was absent due to decalcified processing, we found that Raman peak intensities of amide I and C-C stretching bands were significantly different between OVX and sham-operated specimens. These differences on the Raman spectra were statistically compared by multivariate analyses, principal component analysis (PCA) and liner discrimination analysis (LDA). Our analyses suggest that amide I and C-C stretching bands can be related to stability of bone matrix which reflects bone quality.

Transglutaminase reactivity with gelatine: perspective applications in tissue engineering.

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Bertoni, F; Barbani, N; Giusti, P; Ciardelli, G

2006-05-01

Gelatine was crosslinked by means of an enzymatic treatment using tissue transglutaminase (tTGase) (Sigma) and microbial transglutaminase (mTGase) (Ajinomoto) which catalyses the formation of isopeptide bonds between the gamma-carbonyl group of a glutamine residue and the epsilon-amino group of a lysine residue. The reaction is an interesting alternative to the traditional glutaraldehyde crosslinking, which has several drawbacks (e.g., in medical application) due to the toxicity of the chemical reagent. To further investigate the possibility to utilize the modified protein for tissue engineering application, TGase crosslinked gelatine was incorporated in a gellan matrix, a polysaccharide, to enhance the stability in aqueous media. Films obtained by casting were characterized by thermal analysis, chemical imaging, swelling behaviour and cell adhesion.

Thermal characterisation of gelatin extracted from yellowfin tuna skin and commercial mammalian gelatin.

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Rahman, Mohammad Shafiur; Al-Saidi, Ghalib Said; Guizani, Nejib

2008-05-15

Glass transition and other thermal characteristics of gelatin from different sources were studied by differential scanning calorimetry (DSC) and modulated DSC (MDSC). The initial glass transition temperatures of equilibrated gelatin samples at 11.3% relative humidity, determined from reversible heat flow thermogram of MDSC, were 23, 75 and 59°C, respectively, for tuna skin, bovine and porcine gelatin. When gelatin samples were equilibrated at higher relative humidity of 52.9%, glass transition temperature of fish skin and bovine gelatin decreased to -3 and 57°C, respectively. Further increase of equilibration relative humidity to 75.3% showed increased value in the case of tuna skin, whereas bovine and porcine did not show any significant change. DSC and MDSC results indicated that tuna gelatin showed lower glass transition compared to mammalian source gelatin equilibrated at the same constant relative humidity. In general glass transition measured by DSC was found lower than the values measured by MDSC. The results in this study showed that the degree of plasticization varied with the source of gelatin as well as their extraction methods. Copyright © 2007 Elsevier Ltd. All rights reserved.

Quantitative mapping of matrix content and distribution across the ligament-to-bone insertion.

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Jeffrey P Spalazzi

Full Text Available The interface between bone and connective tissues such as the Anterior Cruciate Ligament (ACL constitutes a complex transition traversing multiple tissue regions, including non-calcified and calcified fibrocartilage, which integrates and enables load transfer between otherwise structurally and functionally distinct tissue types. The objective of this study was to investigate region-dependent changes in collagen, proteoglycan and mineral distribution, as well as collagen orientation, across the ligament-to-bone insertion site using Fourier transform infrared spectroscopic imaging (FTIR-I. Insertion site-related differences in matrix content were also evaluated by comparing tibial and femoral entheses. Both region- and site-related changes were observed. Collagen content was higher in the ligament and bone regions, while decreasing across the fibrocartilage interface. Moreover, interfacial collagen fibrils were aligned parallel to the ligament-bone interface near the ligament region, assuming a more random orientation through the bulk of the interface. Proteoglycan content was uniform on average across the insertion, while its distribution was relatively less variable at the tibial compared to the femoral insertion. Mineral was only detected in the calcified interface region, and its content increased exponentially across the mineralized fibrocartilage region toward bone. In addition to new insights into matrix composition and organization across the complex multi-tissue junction, findings from this study provide critical benchmarks for the regeneration of soft tissue-to-bone interfaces and integrative soft tissue repair.

Quantitative Mapping of Matrix Content and Distribution across the Ligament-to-Bone Insertion

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Spalazzi, Jeffrey P.; Boskey, Adele L.; Pleshko, Nancy; Lu, Helen H.

2013-01-01

The interface between bone and connective tissues such as the Anterior Cruciate Ligament (ACL) constitutes a complex transition traversing multiple tissue regions, including non-calcified and calcified fibrocartilage, which integrates and enables load transfer between otherwise structurally and functionally distinct tissue types. The objective of this study was to investigate region-dependent changes in collagen, proteoglycan and mineral distribution, as well as collagen orientation, across the ligament-to-bone insertion site using Fourier transform infrared spectroscopic imaging (FTIR-I). Insertion site-related differences in matrix content were also evaluated by comparing tibial and femoral entheses. Both region- and site-related changes were observed. Collagen content was higher in the ligament and bone regions, while decreasing across the fibrocartilage interface. Moreover, interfacial collagen fibrils were aligned parallel to the ligament-bone interface near the ligament region, assuming a more random orientation through the bulk of the interface. Proteoglycan content was uniform on average across the insertion, while its distribution was relatively less variable at the tibial compared to the femoral insertion. Mineral was only detected in the calcified interface region, and its content increased exponentially across the mineralized fibrocartilage region toward bone. In addition to new insights into matrix composition and organization across the complex multi-tissue junction, findings from this study provide critical benchmarks for the regeneration of soft tissue-to-bone interfaces and integrative soft tissue repair. PMID:24019964

Demineralized dentin matrix composite collagen material for bone tissue regeneration.

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Li, Jianan; Yang, Juan; Zhong, Xiaozhong; He, Fengrong; Wu, Xiongwen; Shen, Guanxin

2013-01-01

Demineralized dentin matrix (DDM) had been successfully used in clinics as bone repair biomaterial for many years. However, particle morphology of DDM limited it further applications. In this study, DDM and collagen were prepared to DDM composite collagen material. The surface morphology of the material was studied by scanning electron microscope (SEM). MC3T3-E1 cells responses in vitro and tissue responses in vivo by implantation of DDM composite collagen material in bone defect of rabbits were also investigated. SEM analysis showed that DDM composite collagen material evenly distributed and formed a porous scaffold. Cell culture and animal models results indicated that DDM composite collagen material was biocompatible and could support cell proliferation and differentiation. Histological evaluation showed that DDM composite collagen material exhibited good biocompatibility, biodegradability and osteoconductivity with host bone in vivo. The results suggested that DDM composite collagen material might have a significant clinical advantage and potential to be applied in bone and orthopedic surgery.

Two types of mineral-related matrix vesicles in the bone mineralization of zebrafish

International Nuclear Information System (INIS)

Yang, L; Zhang, Y; Cui, F Z

2007-01-01

Two types of mineral-related matrix vesicle, multivesicular body (MVB) and monovesicle, were detected in the skeletal bone of zebrafish. Transmission electron microscopy and energy dispersive spectroscopy (EDS) analyses of the vesicular inclusions reveal that both types of vesicles contain calcium and phosphorus, suggesting that these vesicles may be involved in mineral ion delivery for the bone mineralization of zebrafish. However, their size and substructure are quite different. Monovesicles, whose diameter ranges from 100 nm to 550 nm, are similar to the previously reported normal matrix vesicles, while MVBs have a larger size of 700-1000 nm in nominal diameter and possess a substructure that is composed of smaller vesicles with their average size around 100 nm. The presence of mineral-related MVBs, which is first identified in zebrafish bone, indicates that the mineralization-associated transportation process of mineral ions is more complicated than is ordinarily imagined

Extraction and physicochemical characterization of broiler (Gallus gallus domesticus) skin gelatin compared to commercial bovine gelatin.

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Aykin-Dinçer, E; Koç, A; Erbas, M

2017-09-01

Gelatin was extracted from broiler (Gallus gallus domesticus) skins and analyzed to compare its physicochemical properties with those of commercial bovine gelatin. The average yield of broiler skin gelatin was 6.5% on a wet weight basis. Broiler skin gelatin had more α1-and α2-chains than β-chain and contained high molecular weight (γ-chain) polymers. Glycine was the dominant amino acid in broiler skin gelatin (20.26%), followed by proline (Pro) (15.12%) then hydroxyproline (Hyp) (11.36%). Compared to commercial bovine gelatin, broiler skin gelatin had less total imino acids (Pro and Hyp) but a higher (33.65 vs. 31.38°C) melting temperature (P commercial bovine gelatins appeared to be associated with differences in their amino acid composition and molecular weight distribution. The sensory evaluation results revealed that broiler skin gelatin could be a potential alternative to commercial bovine gelatin, useful in various food products. © 2017 Poultry Science Association Inc.

The role of lysyl oxidase, the extracellular matrix and the pre-metastatic niche in bone metastasis

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Alison Gartland

2016-09-01

Full Text Available Most deaths from solid cancers occur as a result of secondary metastasis to distant sites. Bone is the most frequent metastatic site for many cancer types and can account for up to 80% of cancer-related deaths in certain tumours. The progression from a discrete solid primary tumour to devastating and painful bone metastases is a complex process involving multiple cell types and steps. There is increasing evidence that modulation of the extracellular matrix plays an important role in the lethal transition from a primary to disseminated metastatic bone tumour. This review provides an overview of the current understanding on the role of role of lysyl oxidase, the extracellular matrix and the pre-metastatic niche in bone metastasis

Bioactive glass-ceramic bone repair associated or not with autogenous bone: a study of organic bone matrix organization in a rabbit critical-sized calvarial model.

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Biguetti, Claudia Cristina; Cavalla, Franco; Tim, Carla Roberta; Saraiva, Patrícia Pinto; Orcini, Wilson; De Andrade Holgado, Leandro; Rennó, Ana Claudia Muniz; Matsumoto, Mariza Akemi

2018-04-26

The aim of the study was to analyze bone matrix (BMX) organization after bone grafting and repair using a new bioactive glass-ceramic (Biosilicate ® ) associated or not with particulate autogenous bone graft. Thirty rabbits underwent surgical bilateral parietal defects and divided into groups according to the materials used: (C) control-blood clot, (BG) particulate autogenous bone, (BS) bioactive glass-ceramic, and BG + BS. After 7, 14, and 30 days post-surgery, a fragment of each specimen was fixed in - 80 °C liquid nitrogen for zymographic evaluation, while the remaining was fixed in 10% formalin for histological birefringence analysis. The results of this study demonstrated that matrix organization in experimental groups was significantly improved compared to C considering collagenous organization. Zymographic analysis revealed pro-MMP-2, pro-MMP-9, and active (a)-MMP-2 in all groups, showing gradual decrease of total gelatinolytic activity during the periods. At day 7, BG presented more prominent gelatinolytic activity for pro-MMP-2 and 9 and a-MMP-2, when compared to the other groups. In addition, at day 7, a 53% activation ratio (active form/[active form + latent form]) was evident in C group, 33% in BS group, and 31% in BG group. In general, BS allowed the production of a BMX similar to BG, with organized collagen deposition and MMP-2 and MMP-9 disponibility, permitting satisfactory bone remodeling at the late period. The evaluation of new bone substitute, with favorable biological properties, opens the possibility for its use as a viable and efficient alternative to autologous bone graft.

Soft, chewable gelatin-based pharmaceutical oral formulations: a technical approach.

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Dille, Morten J; Hattrem, Magnus N; Draget, Kurt I

2018-06-01

Hard tablets and capsules for oral drug delivery cause problems for people experiencing dysphagia. This work describes the formulation and properties of a gelatin based, self-preserved, and soft chewable tablet as an alternative and novel drug delivery format. Gelatin (8.8-10% in 24.7-29% water) constituted the matrix of the soft, semi-solid tablets. Three different pharmaceuticals (Ibuprofen 10%, Acetaminophen 15%, and Meloxicam 1.5%) were tested in this formulation. Microbial stability was controlled by lowering the water activity with a mixture of sorbitol and xylitol (45.6-55%). Rheological properties were tested applying small strain oscillation measurements. Taste masking of ibuprofen soft-chew tablets was achieved by keeping the ibuprofen insoluble at pH 4.5 and keeping the processing temperature below the crystalline-to-amorphous transition temperature. Soft-chew formulations showed good stability for all three pharmaceuticals (up to 24 months), and the ibuprofen containing formulation exhibited comparable dissolution to a standard oral tablet as well as good microbial stability. The rheological properties of the ibuprofen/gelatin formulation had the fingerprint of a true gelatin gel, albeit higher moduli, and melting temperature. The results suggest that easy-to-swallow and well taste-masked soft chewable tablet formulations with extended shelf life are within reach for several active pharmaceutical ingredients (APIs).

Gelatin-based laser direct-write technique for the precise spatial patterning of cells.

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Schiele, Nathan R; Chrisey, Douglas B; Corr, David T

2011-03-01

Laser direct-writing provides a method to pattern living cells in vitro, to study various cell-cell interactions, and to build cellular constructs. However, the materials typically used may limit its long-term application. By utilizing gelatin coatings on the print ribbon and growth surface, we developed a new approach for laser cell printing that overcomes the limitations of Matrigel™. Gelatin is free of growth factors and extraneous matrix components that may interfere with cellular processes under investigation. Gelatin-based laser direct-write was able to successfully pattern human dermal fibroblasts with high post-transfer viability (91% ± 3%) and no observed double-strand DNA damage. As seen with atomic force microscopy, gelatin offers a unique benefit in that it is present temporarily to allow cell transfer, but melts and is removed with incubation to reveal the desired application-specific growth surface. This provides unobstructed cellular growth after printing. Monitoring cell location after transfer, we show that melting and removal of gelatin does not affect cellular placement; cells maintained registry within 5.6 ± 2.5 μm to the initial pattern. This study demonstrates the effectiveness of gelatin in laser direct-writing to create spatially precise cell patterns with the potential for applications in tissue engineering, stem cell, and cancer research.

Properties of gelatin-based films incorporated with chitosan-coated microparticles charged with rutin.

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Dammak, Ilyes; Bittante, Ana Mônica Quinta Barbosa; Lourenço, Rodrigo Vinicius; do Amaral Sobral, Paulo José

2017-08-01

The aim of this study was development an active film based on gelatin incorporated with antioxidant, rutin carried into microparticles. The complexation between oppositely charged lecithin and chitosan was applied to prepare the chitosan-coated microparticles. The generated microparticles had an average size of 520±4nm and a span of 0.3 were formulated by a rotor-stator homogenize at the homogenization speed 10,000rpm. Composite films were prepared by incorporating chitosan-coated microparticles, at various concentrations (0.05, 0.1, 0.5, or 1% (based on the weight of the gelatin powder)) in the gelatin-based films. For the prepared films, the results showed that obtained physicochemical, water vapor barrier, and mechanical were compared with native gelatin film with a slight decrease for chitosan concentration higher than 0.5%. The microstructure studies done by scanning electron microscopes, revealed different micropores embedded with oil resulting from the incorporation of the microparticles into the gelatin matrix. Moreover, the calorimetric results were comparable to those of gelatin control film with T g value 45°C and increased crystallinity percentage with increasing incorporation of microparticles. This original concept of composite biodegradable films may thus be a good alternative to incorporate liposoluble active compounds to design an active packaging with good properties. Copyright © 2017 Elsevier B.V. All rights reserved.

Synthesis, characterization and optical properties of gelatin doped with silver nanoparticles

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Mahmoud, K. H.; Abbo, M.

2013-12-01

In this study, silver nanoparticles were synthesized by chemical reduction of silver salt (AgNO3) solution. Formation of nanoparticles was confirmed by UV-visible spectrometry. The surface plasmon resonance peak is located at 430 nm. Doping of silver nanoparticles (Ag NPs) with gelatin biopolymer was studied. The silver content in the polymer matrix was in the range of 0.4-1 wt%. The formation of nanoparticles disappeared for silver content higher than 1 wt%. The morphology and interaction of gelatin doped with Ag NPs was examined by transmission electron microscopy and FTIR spectroscopy. The content of Ag NPs has a pronounced effect on optical and structural properties of gelatin. Optical parameters such as refractive index, complex dielectric constant were calculated. The dispersion of the refractive index was discussed in terms of the single - oscillator Wemple-DiDomenico model. Color properties of the prepared samples were discussed in the framework of CIE L*u*v* color space.

Immobilization of enzymes on radiation-modified gelatine gel by using a chemical cross-linking agent

International Nuclear Information System (INIS)

Bachmann, S.; Gebicka, L.; Galant, S.

1981-01-01

Investigations into the effect of ionizing radiation on the gelatine gels have shown that water-insoluble gel can be formed under suitable irradiation conditions. To establish the optimal conditions for the processing of the insoluble gel, the yield of cross-linking has been determined for gelatine solutions and its gels irradiated with various doses in the absence and in the presence of oxygen. Glucose isomerase (GI) was used as a test enzyme for immobilization on the gelatine gel. This enzyme which catalyses the isomerization of glucose to fructose has been used on the commercial-scale production of high fructose syrups. The support matrix chosen for the enzyme immobilization has been obtained by irradiating 4% wt/vol. de-aerated gelatine gel at a dose of 1.5 x 10 4 kGy at 15 0 C. Actinoplanes missouriensis cells containing GI were mixed with gelatine gel particles and cross-linked with glutaraldehyde. It was found that the immobilized GI can be successfully applied in the continuous isomerization of glucose to fructose. (author)

Formulating gelatin free products

NARCIS (Netherlands)

Buwalda, P.L.

2014-01-01

Gels are applied in many systems in particular in foods. Gelatin is the most common of all gelling agents. In the food industry there is a long quest for replacing gelatin. This chapter focuses on a more application by application approach where a dominant property of gelatin is matched with a

Treatment of osteomyelitis defects by a vancomycin-loaded gelatin/β-tricalcium phosphate composite scaffold

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Zhou, J.; Zhou, X. G.; Wang, J. W.; Zhou, H.; Dong, J.

2018-01-01

Objective In the present study, we aimed to assess whether gelatin/β-tricalcium phosphate (β-TCP) composite porous scaffolds could be used as a local controlled release system for vancomycin. We also investigated the efficiency of the scaffolds in eliminating infections and repairing osteomyelitis defects in rabbits. Methods The gelatin scaffolds containing differing amounts of of β-TCP (0%, 10%, 30% and 50%) were prepared for controlled release of vancomycin and were labelled G-TCP0, G-TCP1, G-TCP3 and G-TCP5, respectively. The Kirby-Bauer method was used to examine the release profile. Chronic osteomyelitis models of rabbits were established. After thorough debridement, the osteomyelitis defects were implanted with the scaffolds. Radiographs and histological examinations were carried out to investigate the efficiency of eliminating infections and repairing bone defects. Results The prepared gelatin/β-TCP scaffolds exhibited a homogeneously interconnected 3D porous structure. The G-TCP0 scaffold exhibited the longest duration of vancomycin release with a release duration of eight weeks. With the increase of β-TCP contents, the release duration of the β-TCP-containing composite scaffolds was decreased. The complete release of vancomycin from the G-TCP5 scaffold was achieved within three weeks. In the treatment of osteomyelitis defects in rabbits, the G-TCP3 scaffold showed the most efficacious performance in eliminating infections and repairing bone defects. Conclusions The composite scaffolds could achieve local therapeutic drug levels over an extended duration. The G-TCP3 scaffold possessed the optimal porosity, interconnection and controlled release performance. Therefore, this scaffold could potentially be used in the treatment of chronic osteomyelitis defects. Cite this article: J. Zhou, X. G. Zhou, J. W. Wang, H. Zhou, J. Dong. Treatment of osteomyelitis defects by a vancomycin-loaded gelatin/β-tricalcium phosphate composite scaffold. Bone Joint Res

Fourier transform infrared imaging microspectroscopy and tissue-level mechanical testing reveal intraspecies variation in mouse bone mineral and matrix composition.

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Courtland, Hayden-William; Nasser, Philip; Goldstone, Andrew B; Spevak, Lyudmila; Boskey, Adele L; Jepsen, Karl J

2008-11-01

Fracture susceptibility is heritable and dependent upon bone morphology and quality. However, studies of bone quality are typically overshadowed by emphasis on bone geometry and bone mineral density. Given that differences in mineral and matrix composition exist in a variety of species, we hypothesized that genetic variation in bone quality and tissue-level mechanical properties would also exist within species. Sixteen-week-old female A/J, C57BL/6J (B6), and C3H/HeJ (C3H) inbred mouse femora were analyzed using Fourier transform infrared imaging and tissue-level mechanical testing for variation in mineral composition, mineral maturity, collagen cross-link ratio, and tissue-level mechanical properties. A/J femora had an increased mineral-to-matrix ratio compared to B6. The C3H mineral-to-matrix ratio was intermediate of A/J and B6. C3H femora had reduced acid phosphate and carbonate levels and an increased collagen cross-link ratio compared to A/J and B6. Modulus values paralleled mineral-to-matrix values, with A/J femora being the most stiff, B6 being the least stiff, and C3H having intermediate stiffness. In addition, work-to-failure varied among the strains, with the highly mineralized and brittle A/J femora performing the least amount of work-to-failure. Inbred mice are therefore able to differentially modulate the composition of their bone mineral and the maturity of their bone matrix in conjunction with tissue-level mechanical properties. These results suggest that specific combinations of bone quality and morphological traits are genetically regulated such that mechanically functional bones can be constructed in different ways.

Injection of Unicameral Bone Cysts with Bone Marrow Aspirate and Demineralized Bone Matrix Avoids Open Curettage and Bone Grafting in a Retrospective Cohort.

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Gundle, Kenneth R; Bhatt, Etasha M; Punt, Stephanie E; Bompadre, Viviana; Conrad, Ernest U

2017-01-01

Many treatment options exist for unicameral bone cysts (UBC), without clear evidence of superiority. Meta-analyses have been limited by small numbers of patients in specific anatomic and treatment subgroups. The purpose of this study was to report the outcomes of injecting bone marrow aspirate and demineralized bone matrix (BMA/DBM) for the treatment of proximal humerus UBC. Fifty-one patients with proximal humerus lesions treated by BMA/DBM injection were retrospectively reviewed from a single academic medical center. The mean number of injections performed per patient was 2.14 (range 1-5). Eleven patients underwent only one injection (22%), an additional 19 patients completed treatment after two injections (37%), four patients healed after three injections (8%), and one patient healed after four injections (2%). The cumulative success rate of serial BMA/DBM injections was 22% (11/51), 58% (30/51), 67% (34/51), and 69% (35/51). Eleven patients (22%) ultimately underwent open curettage and bone grafting, and five patients (10%) were treated with injection of calcium phosphate bone substitute. A BMA/DBM injection strategy avoided an open procedure in 78% of patients with a proximal humerus UBC. The majority of patients underwent at least 2 injection treatments. Level IV retrospective cohort study.

Response of induced bone defects in horses to collagen matrix containing the human parathyroid hormone gene.

Science.gov (United States)

Backstrom, Kristin C; Bertone, Alicia L; Wisner, Erik R; Weisbrode, Stephen E

2004-09-01

To determine whether human parathyroid hormone (hPTH) gene in collagen matrix could safely promote bone formation in diaphyseal or subchondral bones of horses. 8 clinically normal adult horses. Amount, rate, and quality of bone healing for 13 weeks were determined by use of radiography, quantitative computed tomography, and histomorphometric analysis. Diaphyseal cortex and subchondral bone defects of metacarpi were filled with hPTH(1-34) gene-activated matrix (GAM) or remained untreated. Joints were assessed on the basis of circumference, synovial fluid analysis, pain on flexion, lameness, and gross and histologic examination. Bone volume index was greater for cortical defects treated with hPTH(1-34) GAM, compared with untreated defects. Bone production in cortical defects treated with hPTH(1-34) GAM positively correlated with native bone formation in untreated defects. In contrast, less bone was detected in hPTH(1-34) GAM-treated subchondral bone defects, compared with untreated defects, and histology confirmed poorer healing and residual collagen sponge. Use of hPTH(1-34) GAM induced greater total bone, specifically periosteal bone, after 13 weeks of healing in cortical defects of horses. The hPTH(1-34) GAM impeded healing of subchondral bone but was biocompatible with joint tissues. Promotion of periosteal bone formation may be beneficial for healing of cortical fractures in horses, but the delay in onset of bone formation may negate benefits. The hPTH(1-34) GAM used in this study should not be placed in articular subchondral bone defects, but contact with articular surfaces is unlikely to cause short-term adverse effects.

Virus immobilization on biomaterial scaffolds through biotin-avidin interaction for improving bone regeneration.

Science.gov (United States)

Hu, Wei-Wen; Wang, Zhuo; Krebsbach, Paul H

2016-02-01

To spatially control therapeutic gene delivery for potential tissue engineering applications, a biotin-avidin interaction strategy was applied to immobilize viral vectors on biomaterial scaffolds. Both adenoviral vectors and gelatin sponges were biotinylated and avidin was applied to link them in a virus-biotin-avidin-biotin-material (VBABM) arrangement. The tethered viral particles were stably maintained within scaffolds and SEM images illustrated that viral particles were evenly distributed in three-dimensional (3D) gelatin sponges. An in vivo study demonstrated that transgene expression was restricted to the implant sites only and transduction efficiency was improved using this conjugation method. For an orthotopic bone regeneration model, adenovirus encoding BMP-2 (AdBMP2) was immobilized to gelatin sponges before implanting into critical-sized bone defects in rat calvaria. Compared to gelatin sponges with AdBMP2 loaded in a freely suspended form, the VBABM method enhanced gene transfer and bone regeneration was significantly improved. These results suggest that biotin-avidin immobilization of viral vectors to biomaterial scaffolds may be an effective strategy to facilitate tissue regeneration. Copyright © 2013 John Wiley & Sons, Ltd.

Laminated electrospun nHA/PHB-composite scaffolds mimicking bone extracellular matrix for bone tissue engineering.

Science.gov (United States)

Chen, Zhuoyue; Song, Yue; Zhang, Jing; Liu, Wei; Cui, Jihong; Li, Hongmin; Chen, Fulin

2017-03-01

Electrospinning is an effective means to generate nano- to micro-scale polymer fibers resembling native extracellular matrix for tissue engineering. However, a major problem of electrospun materials is that limited pore size and porosity may prevent adequate cellular infiltration and tissue ingrowth. In this study, we first prepared thin layers of hydroxyapatite nanoparticle (nHA)/poly-hydroxybutyrate (PHB) via electrospinning. We then laminated the nHA/PHB thin layers to obtain a scaffold for cell seeding and bone tissue engineering. The results demonstrated that the laminated scaffold possessed optimized cell-loading capacity. Bone marrow mesenchymal stem cells (MSCs) exhibited better adherence, proliferation and osteogenic phenotypes on nHA/PHB scaffolds than on PHB scaffolds. Thereafter, we seeded MSCs onto nHA/PHB scaffolds to fabricate bone grafts. Histological observation showed osteoid tissue formation throughout the scaffold, with most of the scaffold absorbed in the specimens 2months after implantation, and blood vessels ingrowth into the graft could be observed in the graft. We concluded that electrospun and laminated nanoscaled biocomposite scaffolds hold great therapeutic potential for bone regeneration. Copyright © 2016 Elsevier B.V. All rights reserved.

Gelatin/chitosan biofilm: preparation and characterization

International Nuclear Information System (INIS)

Trindade, Luciane da C.; Nunes, Raquel A.; Diniz, Nadie K.S.; Braga, Carla R.C.; Silva, Suedina M. de Lima

2011-01-01

In this study, gelatin, chitosan and gelatin/chitosan bio films using the ratio of gelatin/chitosan (50/50) were prepared by casting method. The bio films prepared were characterized by X-ray diffraction, scanning electron microscopy and dissolution ratio. According to the results, the incorporation of chitosan into gelatin indicate the decrease of crystallinity of chitosan, a compact structure without large pores and that the dissolution of gelatin/chitosan film is little influenced by hot water than gelatin films. (author)

Multiscale, Converging Defects of Macro-Porosity, Microstructure and Matrix Mineralization Impact Long Bone Fragility in NF1

Science.gov (United States)

Kühnisch, Jirko; Seto, Jong; Lange, Claudia; Schrof, Susanne; Stumpp, Sabine; Kobus, Karolina; Grohmann, Julia; Kossler, Nadine; Varga, Peter; Osswald, Monika; Emmerich, Denise; Tinschert, Sigrid; Thielemann, Falk; Duda, Georg; Seifert, Wenke; el Khassawna, Thaqif; Stevenson, David A.; Elefteriou, Florent; Kornak, Uwe; Raum, Kay; Fratzl, Peter; Mundlos, Stefan; Kolanczyk, Mateusz

2014-01-01

Bone fragility due to osteopenia, osteoporosis or debilitating focal skeletal dysplasias is a frequent observation in the Mendelian disease Neurofibromatosis type 1 (NF1). To determine the mechanisms underlying bone fragility in NF1 we analyzed two conditional mouse models, Nf1Prx1 (limb knock-out) and Nf1Col1 (osteoblast specific knock-out), as well as cortical bone samples from individuals with NF1. We examined mouse bone tissue with micro-computed tomography, qualitative and quantitative histology, mechanical tensile analysis, small-angle X-ray scattering (SAXS), energy dispersive X-ray spectroscopy (EDX), and scanning acoustic microscopy (SAM). In cortical bone of Nf1Prx1 mice we detected ectopic blood vessels that were associated with diaphyseal mineralization defects. Defective mineral binding in the proximity of blood vessels was most likely due to impaired bone collagen formation, as these areas were completely devoid of acidic matrix proteins and contained thin collagen fibers. Additionally, we found significantly reduced mechanical strength of the bone material, which was partially caused by increased osteocyte volume. Consistent with these observations, bone samples from individuals with NF1 and tibial dysplasia showed increased osteocyte lacuna volume. Reduced mechanical properties were associated with diminished matrix stiffness, as determined by SAM. In line with these observations, bone tissue from individuals with NF1 and tibial dysplasia showed heterogeneous mineralization and reduced collagen fiber thickness and packaging. Collectively, the data indicate that bone fragility in NF1 tibial dysplasia is partly due to an increased osteocyte-related micro-porosity, hypomineralization, a generalized defect of organic matrix formation, exacerbated in the regions of tensional and bending force integration, and finally persistence of ectopic blood vessels associated with localized macro-porotic bone lesions. PMID:24465906

Laminated electrospun nHA/PHB-composite scaffolds mimicking bone extracellular matrix for bone tissue engineering

Energy Technology Data Exchange (ETDEWEB)

Chen, Zhuoyue [Lab of Tissue Engineering, Faculty of Life Science, Northwest University, 229 TaiBai North Road, Xi' an, Shaanxi Province 710069 (China); Provincial Key Laboratory of Biotechnology of Shaanxi, Northwest University, 229 TaiBai North Road, Xi' an, Shaanxi Province 710069 (China); Song, Yue [Lab of Tissue Engineering, Faculty of Life Science, Northwest University, 229 TaiBai North Road, Xi' an, Shaanxi Province 710069 (China); Zhang, Jing [Lab of Tissue Engineering, Faculty of Life Science, Northwest University, 229 TaiBai North Road, Xi' an, Shaanxi Province 710069 (China); Provincial Key Laboratory of Biotechnology of Shaanxi, Northwest University, 229 TaiBai North Road, Xi' an, Shaanxi Province 710069 (China); Key Laboratory of Resource Biology and Modern Biotechnology in Western China, Ministry of Education, Northwest University, 229 TaiBai North Road, Xi' an, Shaanxi Province, 710069 (China); Liu, Wei [Lab of Tissue Engineering, Faculty of Life Science, Northwest University, 229 TaiBai North Road, Xi' an, Shaanxi Province 710069 (China); Cui, Jihong, E-mail: cjh@nwu.edu.cn [Lab of Tissue Engineering, Faculty of Life Science, Northwest University, 229 TaiBai North Road, Xi' an, Shaanxi Province 710069 (China); Provincial Key Laboratory of Biotechnology of Shaanxi, Northwest University, 229 TaiBai North Road, Xi' an, Shaanxi Province 710069 (China); Key Laboratory of Resource Biology and Modern Biotechnology in Western China, Ministry of Education, Northwest University, 229 TaiBai North Road, Xi' an, Shaanxi Province, 710069 (China); and others

2017-03-01

Electrospinning is an effective means to generate nano- to micro-scale polymer fibers resembling native extracellular matrix for tissue engineering. However, a major problem of electrospun materials is that limited pore size and porosity may prevent adequate cellular infiltration and tissue ingrowth. In this study, we first prepared thin layers of hydroxyapatite nanoparticle (nHA)/poly-hydroxybutyrate (PHB) via electrospinning. We then laminated the nHA/PHB thin layers to obtain a scaffold for cell seeding and bone tissue engineering. The results demonstrated that the laminated scaffold possessed optimized cell-loading capacity. Bone marrow mesenchymal stem cells (MSCs) exhibited better adherence, proliferation and osteogenic phenotypes on nHA/PHB scaffolds than on PHB scaffolds. Thereafter, we seeded MSCs onto nHA/PHB scaffolds to fabricate bone grafts. Histological observation showed osteoid tissue formation throughout the scaffold, with most of the scaffold absorbed in the specimens 2 months after implantation, and blood vessels ingrowth into the graft could be observed in the graft. We concluded that electrospun and laminated nanoscaled biocomposite scaffolds hold great therapeutic potential for bone regeneration. - Highlights: • We laminated the nHA/PHB layers to obtain a scaffold for bone tissue engineering. • The laminated scaffold performed optimized cell-loading capacity. • MSCs exhibited osteogenic phenotypes on the laminated scaffold. • Osteoid tissue formed throughout the laminated scaffold after 2 months in vivo. The laminated bio-composite scaffolds can be applied to bone regeneration.

Laminated electrospun nHA/PHB-composite scaffolds mimicking bone extracellular matrix for bone tissue engineering

International Nuclear Information System (INIS)

Chen, Zhuoyue; Song, Yue; Zhang, Jing; Liu, Wei; Cui, Jihong

2017-01-01

Electrospinning is an effective means to generate nano- to micro-scale polymer fibers resembling native extracellular matrix for tissue engineering. However, a major problem of electrospun materials is that limited pore size and porosity may prevent adequate cellular infiltration and tissue ingrowth. In this study, we first prepared thin layers of hydroxyapatite nanoparticle (nHA)/poly-hydroxybutyrate (PHB) via electrospinning. We then laminated the nHA/PHB thin layers to obtain a scaffold for cell seeding and bone tissue engineering. The results demonstrated that the laminated scaffold possessed optimized cell-loading capacity. Bone marrow mesenchymal stem cells (MSCs) exhibited better adherence, proliferation and osteogenic phenotypes on nHA/PHB scaffolds than on PHB scaffolds. Thereafter, we seeded MSCs onto nHA/PHB scaffolds to fabricate bone grafts. Histological observation showed osteoid tissue formation throughout the scaffold, with most of the scaffold absorbed in the specimens 2 months after implantation, and blood vessels ingrowth into the graft could be observed in the graft. We concluded that electrospun and laminated nanoscaled biocomposite scaffolds hold great therapeutic potential for bone regeneration. - Highlights: • We laminated the nHA/PHB layers to obtain a scaffold for bone tissue engineering. • The laminated scaffold performed optimized cell-loading capacity. • MSCs exhibited osteogenic phenotypes on the laminated scaffold. • Osteoid tissue formed throughout the laminated scaffold after 2 months in vivo. The laminated bio-composite scaffolds can be applied to bone regeneration.

Incorporation of mesoporous silica nanoparticles into random electrospun PLGA and PLGA/gelatin nanofibrous scaffolds enhances mechanical and cell proliferation properties

International Nuclear Information System (INIS)

Mehrasa, Mohammad; Asadollahi, Mohammad Ali; Nasri-Nasrabadi, Bijan; Ghaedi, Kamran; Salehi, Hossein; Dolatshahi-Pirouz, Alireza; Arpanaei, Ayyoob

2016-01-01

Poly(lactic-co-glycolic acid) (PLGA) and PLGA/gelatin random nanofibrous scaffolds embedded with different amounts of mesoporous silica nanoparticles (MSNPs) were fabricated using electrospinning method. To evaluate the effects of nanoparticles on the scaffolds, physical, chemical, and mechanical properties as well as in vitro degradation behavior of scaffolds were investigated. The mean diameters of nanofibers were 974 ± 68 nm for the pure PLGA scaffolds vs 832 ± 70, 764 ± 80, and 486 ± 64 for the PLGA/gelatin, PLGA/10 wt% MSNPs, and the PLGA/gelatin/10 wt% MSNPs scaffolds, respectively. The results suggested that the incorporation of gelatin and MSNPs into PLGA-based scaffolds enhances the hydrophilicity of scaffolds due to an increase of hydrophilic functional groups on the surface of nanofibers. With porosity examination, it was concluded that the incorporation of MSNPs and gelatin decrease the porosity of scaffolds. Nanoparticles also improved the tensile mechanical properties of scaffolds. Using in vitro degradation analysis, it was shown that the addition of nanoparticles to the nanofibers matrix increases the weight loss percentage of PLGA-based samples, whereas it decreases the weight loss percentage in the PLGA/gelatin composites. Cultivation of rat pheochromocytoma cell line (PC12), as precursor cells of dopaminergic neural cells, on the scaffolds demonstrated that the introduction of MSNPs into PLGA and PLGA/gelatin matrix leads to improved cell attachment and proliferation and enhances cellular processes. - Highlights: • PLGA-based random nanofibers embedded with mesoporous silica nanoparticles were fabricated using electrospinning method • Incorporation of gelatin and MSNPs into PLGA-based scaffolds increased the hydrophilicity of scaffold • Addition of nanoparticles also improved the tensile mechanical properties of scaffolds • Introduction of MSNPs led to improved cell attachment and proliferation

Incorporation of mesoporous silica nanoparticles into random electrospun PLGA and PLGA/gelatin nanofibrous scaffolds enhances mechanical and cell proliferation properties

Energy Technology Data Exchange (ETDEWEB)

Mehrasa, Mohammad [Department of Biotechnology, Faculty of Advanced Sciences and Technologies, University of Isfahan, Isfahan 81746-73441 (Iran, Islamic Republic of); Department of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran (Iran, Islamic Republic of); Asadollahi, Mohammad Ali, E-mail: ma.asadollahi@ast.ui.ac.ir [Department of Biotechnology, Faculty of Advanced Sciences and Technologies, University of Isfahan, Isfahan 81746-73441 (Iran, Islamic Republic of); Nasri-Nasrabadi, Bijan [Department of Chemical Engineering, Isfahan University of Technology, Isfahan (Iran, Islamic Republic of); Ghaedi, Kamran [Department of Biology, Faculty of Science, University of Isfahan, Isfahan 81746-73441 (Iran, Islamic Republic of); Salehi, Hossein [Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan (Iran, Islamic Republic of); Dolatshahi-Pirouz, Alireza [DTU Nanotech, Center for Nanomedicine and Theranostics, Technical University of Denmark (DTU), DK-2800 Kgs. Lyngby (Denmark); Arpanaei, Ayyoob, E-mail: arpanaei@yahoo.com [Department of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran (Iran, Islamic Republic of)

2016-09-01

Poly(lactic-co-glycolic acid) (PLGA) and PLGA/gelatin random nanofibrous scaffolds embedded with different amounts of mesoporous silica nanoparticles (MSNPs) were fabricated using electrospinning method. To evaluate the effects of nanoparticles on the scaffolds, physical, chemical, and mechanical properties as well as in vitro degradation behavior of scaffolds were investigated. The mean diameters of nanofibers were 974 ± 68 nm for the pure PLGA scaffolds vs 832 ± 70, 764 ± 80, and 486 ± 64 for the PLGA/gelatin, PLGA/10 wt% MSNPs, and the PLGA/gelatin/10 wt% MSNPs scaffolds, respectively. The results suggested that the incorporation of gelatin and MSNPs into PLGA-based scaffolds enhances the hydrophilicity of scaffolds due to an increase of hydrophilic functional groups on the surface of nanofibers. With porosity examination, it was concluded that the incorporation of MSNPs and gelatin decrease the porosity of scaffolds. Nanoparticles also improved the tensile mechanical properties of scaffolds. Using in vitro degradation analysis, it was shown that the addition of nanoparticles to the nanofibers matrix increases the weight loss percentage of PLGA-based samples, whereas it decreases the weight loss percentage in the PLGA/gelatin composites. Cultivation of rat pheochromocytoma cell line (PC12), as precursor cells of dopaminergic neural cells, on the scaffolds demonstrated that the introduction of MSNPs into PLGA and PLGA/gelatin matrix leads to improved cell attachment and proliferation and enhances cellular processes. - Highlights: • PLGA-based random nanofibers embedded with mesoporous silica nanoparticles were fabricated using electrospinning method • Incorporation of gelatin and MSNPs into PLGA-based scaffolds increased the hydrophilicity of scaffold • Addition of nanoparticles also improved the tensile mechanical properties of scaffolds • Introduction of MSNPs led to improved cell attachment and proliferation.

Aluminum and bone: Review of new clinical circumstances associated with Al(3+) deposition in the calcified matrix of bone.

Science.gov (United States)

Chappard, D; Bizot, P; Mabilleau, G; Hubert, L

2016-06-01

Several decades ago, aluminum encephalopathy associated with osteomalacia has been recognized as the major complication of chronic renal failure in dialyzed patients. Removal of aluminum from the dialysate has led to a disappearance of the disease. However, aluminum deposit occurs in the hydroxyapatite of the bone matrix in some clinical circumstances that are presented in this review. We have encountered aluminum in bone in patients with an increased intestinal permeability (coeliac disease), or in the case of prolonged administration of aluminum anti-acid drugs. A colocalisation of aluminum with iron was also noted in cases of hemochromatosis and sickle cell anemia. Aluminium was also identified in a series of patients with exostosis, a frequent benign bone tumor. Corrosion of prosthetic implants composed of grade V titanium (TA6V is an alloy containing 6% aluminum and 4% vanadium) was also observed in a series of hip or knee revisions. Aluminum can be identified in undecalcified bone matrix stained by solochrome azurine, a highly specific stain allowing the detection of 0.03 atomic %. Colocalization of aluminum and iron does not seem to be the fruit of chance but the cellular and molecular mechanisms are still poorly understood. Histochemistry is superior to spectroscopic analyses (EDS and WDS in scanning electron microscopy). Copyright © 2015 Elsevier Masson SAS. All rights reserved.

EKSTRAKSI GELATIN DARI KAKI AYAM BROILER MELALUI BERBAGAI LARUTAN ASAM DAN BASA DENGAN VARIASI LAMA PERENDAMAN

Directory of Open Access Journals (Sweden)

Muhammad Rasyid Indrawan

2016-12-01

Full Text Available Gelatin is a biopolymer that can be obtained from partially hydrolysis of collagen present in skin, bone, and connective tissues of animals This study used chicken Broiler feet as a source of collagen. This study was conducted to determine the effect of soaking treatment with solution of acid and base, and with variety of soaking time. The solution used is HCl, CH3COOH, and NaOH with different concentration. The process of soaking followed by extraction, filtration, and drying to obtain a sheet of gelatin. Gelatin was analyzed qualitatively with chemical reaction. The best results of yield can be obtained from various methods of soaking given by HCl 2% for 2 days, CH3COOH 2% for 3 days, and NaOH 2% for 1 day. Keywords : Gelatin, Chicken feet Broiler’s, Acid-treated, Alkali-treated ABSTRAK Gelatin merupakan biopolimer yang biasanya diperoleh dari hidrolisis parsial kolagen jaringan kulit, tulang, dan jaringan ikat hewan. Penelitian ini menggunakan kaki ayam broiler sebagai sumber kolagen. Penelitian dilakukan untuk melihat pengaruh perlakuan perendaman kaki ayam broiler melalui berbagai larutan asam dan basa dengan variasi lama perendaman. Larutan yang digunakan yaitu HCl, CH3COOH, dan NaOH dengan berbagai konsentrasi. Proses perendaman dilanjutkan dengan ekstraksi, filtrasi, dan pengeringan untuk mendapatkan lembaran gelatin. Gelatin yang diperoleh dianalisis secara kualitatif melalui reaksi kimia. Hasil rendemen terbaik dapat diperoleh dari berbagai metode perendaman yaitu HCl 2% selama 2 hari, CH3COOH 2% selama 3 hari, dan NaOH 2% selama 1 hari. Kata Kunci : Gelatin, Kaki ayam Broiler, Metode asam, Metode basa

Ultra-structural defects cause low bone matrix stiffness despite high mineralization in osteogenesis imperfecta mice.

Science.gov (United States)

Vanleene, Maximilien; Porter, Alexandra; Guillot, Pascale-Valerie; Boyde, Alan; Oyen, Michelle; Shefelbine, Sandra

2012-06-01

Bone is a complex material with a hierarchical multi-scale organization from the molecule to the organ scale. The genetic bone disease, osteogenesis imperfecta, is primarily caused by mutations in the collagen type I genes, resulting in bone fragility. Because the basis of the disease is molecular with ramifications at the whole bone level, it provides a platform for investigating the relationship between structure, composition, and mechanics throughout the hierarchy. Prior studies have individually shown that OI leads to: 1. increased bone mineralization, 2. decreased elastic modulus, and 3. smaller apatite crystal size. However, these have not been studied together and the mechanism for how mineral structure influences tissue mechanics has not been identified. This lack of understanding inhibits the development of more accurate models and therapies. To address this research gap, we used a mouse model of the disease (oim) to measure these outcomes together in order to propose an underlying mechanism for the changes in properties. Our main finding was that despite increased mineralization, oim bones have lower stiffness that may result from the poorly organized mineral matrix with significantly smaller, highly packed and disoriented apatite crystals. Using a composite framework, we interpret the lower oim bone matrix elasticity observed as the result of a change in the aspect ratio of apatite crystals and a disruption of the crystal connectivity. Copyright © 2012 Elsevier Inc. All rights reserved.

How to exhaust your bone marrow

DEFF Research Database (Denmark)

Salomo, Louise; Salomo, Morten; Andersen, Steven A W

2013-01-01

at work and in his spare time, and kept a very thorough training and weight diary. Owing to a high intake of energy and protein drinks he tried to optimise his physical performance and kept a normal body mass index  at 23.7. A bone marrow biopsy showed gelatinous bone marrow transformation, normally seen...

Calcium-phosphate matrix with or without TGF-β3 improves tendon-bone healing after rotator cuff repair.

Science.gov (United States)

Kovacevic, David; Fox, Alice J; Bedi, Asheesh; Ying, Liang; Deng, Xiang-Hua; Warren, Russell F; Rodeo, Scott A

2011-04-01

Rotator cuff tendon heals by formation of an interposed zone of fibrovascular scar tissue. Recent studies demonstrate that transforming growth factor-beta 3 (TGF-β(3)) is associated with tissue regeneration and "scarless" healing, in contrast to scar-mediated healing that occurs with TGF-β(1). Delivery of TGF-β(3) in an injectable calcium-phosphate matrix to the healing tendon-bone interface after rotator cuff repair will result in increased attachment strength secondary to improved bone formation and collagen organization and reduced scar formation of the healing enthesis. Controlled laboratory study. Ninety-six male Sprague-Dawley rats underwent unilateral detachment of the supraspinatus tendon followed by acute repair using transosseous suture fixation. Animals were allocated into 1 of 3 groups: (1) repair alone (controls, n = 32), (2) repair augmented by application of an osteoconductive calcium-phosphate (Ca-P) matrix only (n = 32), or (3) repair augmented with Ca-P matrix + TGF-β(3) (2.75 µg) at the tendon-bone interface (n = 32). Animals were euthanized at either 2 weeks or 4 weeks postoperatively. Biomechanical testing of the supraspinatus tendon-bone complex was performed at 2 and 4 weeks (n = 8 per group). Microcomputed tomography was utilized to quantitate bone microstructure at the repair site. The healing tendon-bone interface was evaluated with histomorphometry and immunohistochemical localization of collagen types I (COLI) and III (COLIII). Statistical analysis was performed using 2-way analysis of variance with significance set at P repair site is associated with new bone formation, increased fibrocartilage, and improved collagen organization at the healing tendon-bone interface in the early postoperative period after rotator cuff repair. The addition of TGF-β(3) significantly improved strength of the repair at 4 weeks postoperatively and resulted in a more favorable COLI/COLIII ratio. The delivery of TGF-β(3) with an injectable Ca-P matrix

Mechanical and barrier properties of maize starch-gelatin composite films: effects of amylose content.

Science.gov (United States)

Wang, Kun; Wang, Wenhang; Ye, Ran; Xiao, Jingdong; Liu, Yaowei; Ding, Junsheng; Zhang, Shaojing; Liu, Anjun

2017-08-01

In order to obtain new reinforcing bio-fillers to improve the physicochemical properties of gelatin-based films, three types of maize starch, waxy maize starch (Ap), normal starch (Ns) and high-amylose starch (Al), were incorporated into gelatin film and the resulting film properties were investigated, focusing on the impact of amylose content. The thickness, opacity and roughness of gelatin film increased depending on the amylose content along with the starch concentration. The effects of the three starches on the mechanical properties of gelatin film were governed by amylose content, starch concentration as well as environmental relative humidity (RH). At 75% RH, the presence of Al and Ns in the gelatin matrix increased the film strength but decreased its elongation, while Ap exhibited an inverse effect. Starch addition decreased the oxygen permeability of the film, with the lowest value at 20% Al and Ns. All starches, notably at 30% content, led to a decrease in the water vapor permeability of the film at 90% RH, especially Ns starch. Furthermore, the starches improved the thermal stability of the film to some extent. Fourier transform infrared spectra indicated that some weak intermolecular interactions such as hydrogen bonding occurred between gelatin and starch. Moreover, a high degree of B-type crystallinity of starch was characterized in Gel-Al film by X-ray diffraction. Tailoring the properties of gelatin film by the incorporation of different types of maize starch provides the potential to extend its applications in edible food packaging. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

On spray drying of oxidized corn starch cross-linked gelatin microcapsules for drug release

International Nuclear Information System (INIS)

Dang, Xugang; Yang, Mao; Shan, Zhihua; Mansouri, Shahnaz; May, Bee K; Chen, Xiaodong; Chen, Hui; Woo, Meng Wai

2017-01-01

Spray-dried gelatin/oxidized corn starch (G/OCS) microcapsules were produced for drug release application. The prepared microcapsules were characterized through a scanning electron microscope (SEM) picture and thermogravimetric analysis (TGA). The swelling characteristics of the G/OCS microcapsules and release properties of vitamin C were then investigated. The results from structural analysis indicated that the presence of miscibility and compatibility between oxidized corn starch and gelatin, and exhibits high thermal stability up to 326 °C. The swelling of G/OCS microcapsules increased with increasing pH and reduced with decreasing ionic strength, attributed to the cross-linking between gelatin and oxidized corn starch, ionization of functional groups. Vitamin C release characteristic revealed controlled release behavior in the first 3 h of contact with an aqueous medium. This release behavior was independent of the swelling behavior indicating the potential of the encapsulating matrix to produce controlled release across a spectrum of pH environment. - Highlights: • It's first time to prepare microencapsulation with gelatin and oxidized corn starch. • The microencapsulation material can be biodegradable completely. • The production technology of microcapsule is convenient. • This work explores the potential to use oxidized starch cross-linked gelatin. • The microencapsulation material can be used for drug release.

On spray drying of oxidized corn starch cross-linked gelatin microcapsules for drug release

Energy Technology Data Exchange (ETDEWEB)

Dang, Xugang; Yang, Mao; Shan, Zhihua [National Engineering Laboratory for Clean Technology Leather Manufacture, Sichuan University, Chengdu, Sichuan 610065 (China); Mansouri, Shahnaz [Department of Chemical Engineering, Monash University, VIC 3800 (Australia); May, Bee K [School of Applied Science, RMIT University, 124 La Trobe St, Melbourne, VIC 3001 (Australia); Chen, Xiaodong [Department of Chemical Engineering, Monash University, VIC 3800 (Australia); School of Chemical and Environmental Engineering, College of Chemistry, Chemical Engineering and Material Science, Soochow University (China); Chen, Hui, E-mail: leather2088@sina.com [National Engineering Laboratory for Clean Technology Leather Manufacture, Sichuan University, Chengdu, Sichuan 610065 (China); Department of Chemical Engineering, Monash University, VIC 3800 (Australia); Woo, Meng Wai, E-mail: meng.woo@monash.edu [Department of Chemical Engineering, Monash University, VIC 3800 (Australia)

2017-05-01

Spray-dried gelatin/oxidized corn starch (G/OCS) microcapsules were produced for drug release application. The prepared microcapsules were characterized through a scanning electron microscope (SEM) picture and thermogravimetric analysis (TGA). The swelling characteristics of the G/OCS microcapsules and release properties of vitamin C were then investigated. The results from structural analysis indicated that the presence of miscibility and compatibility between oxidized corn starch and gelatin, and exhibits high thermal stability up to 326 °C. The swelling of G/OCS microcapsules increased with increasing pH and reduced with decreasing ionic strength, attributed to the cross-linking between gelatin and oxidized corn starch, ionization of functional groups. Vitamin C release characteristic revealed controlled release behavior in the first 3 h of contact with an aqueous medium. This release behavior was independent of the swelling behavior indicating the potential of the encapsulating matrix to produce controlled release across a spectrum of pH environment. - Highlights: • It's first time to prepare microencapsulation with gelatin and oxidized corn starch. • The microencapsulation material can be biodegradable completely. • The production technology of microcapsule is convenient. • This work explores the potential to use oxidized starch cross-linked gelatin. • The microencapsulation material can be used for drug release.

Demineralized bone matrix and human cancellous bone enhance fixation of porous-coated titanium implants in sheep

DEFF Research Database (Denmark)

Babiker, Hassan; Ding, Ming; Overgaard, Søren

2016-01-01

matrix (DBM), alone or in combination with allograft or commercially available human cancellous bone (CB), may replace allografts, as they have the capability of inducing new bone and improving implant fixation through enhancing bone ongrowth. The purpose of this study was to investigate the effect...... of DBM alone, DBM with CB, or allograft on the fixation of porous-coated titanium implants. DBM100 and CB produced from human tissue were included. Both materials are commercially available. DBM granules are placed in pure DBM and do not contain any other carrier. Titanium alloy implants, 10 mm long × 10...... mm diameter, were inserted bilaterally into the femoral condyles of eight skeletally mature sheep. Thus, four implants with a concentric gap of 2 mm were implanted in each sheep. The gap was filled with: (a) DBM; (b) DBM:CB at a ratio of 1:3; (c) DBM:allograft at a ratio of 1:3; or (d) allograft...

Castor oil polymer induces bone formation with high matrix metalloproteinase-2 expression.

Science.gov (United States)

Saran, Wallace Rocha; Chierice, Gilberto Orivaldo; da Silva, Raquel Assed Bezerra; de Queiroz, Alexandra Mussolino; Paula-Silva, Francisco Wanderley Garcia; da Silva, Léa Assed Bezerra

2014-02-01

The aim of this study was to evaluate the modulation of matrix metalloproteinase-2 (MMP-2) and -9 (MMP-9) expression in newly formed bone tissue at the interface between implants derived from castor oil (Ricinus communis) polymer and the tibia medullary canal. Forty-four rabbits were assigned to either Group 1 (n = 12; control) or Group 2 (n = 30), which had the tibial medullary canals reamed bilaterally and filled with polymer. CT scans showed no space between the material surface and the bone at the implant/bone marrow interface, and the density of the tissues at this interface was similar to the density measured of other regions of the bone. At 90 days postimplantation, the interface with the polymer presented a thick layer of newly formed bone tissue rich in osteocytes. This tissue exhibited ongoing maturation at 120 and 150 days postimplantation. Overall, bone remodeling process was accompanied by positive modulation of MMP-2 and low MMP-9 expression. Differently, in control group, the internal surface close to the medullary canal was lined by osteoblasts, followed by a bone tissue zone with few lacunae filled with osteocytes. Maturation of the tissue of the medullary internal surface occurred in the inner region, with the bone being nonlamellar. © 2013 Wiley Periodicals, Inc.

Analysing the bioactive makeup of demineralised dentine matrix on bone marrow mesenchymal stem cells for enhanced bone repair.

Science.gov (United States)

Avery, S J; Sadaghiani, L; Sloan, A J; Waddington, R J

2017-07-10

Dentine matrix has proposed roles for directing mineralised tissue repair in dentine and bone; however, the range of bioactive components in dentine and specific biological effects on bone-derived mesenchymal stem cells (MSCs) in humans are less well understood. The aims of this study were to further elucidate the biological response of MSCs to demineralised dentine matrix (DDM) in enhancing wound repair responses and ascertain key contributing components. Dentine was obtained from human teeth and DDM proteins solubilised with ethylenediaminetetraacetic acid (EDTA). Bone marrow derived MSCs were commercially obtained. Cells with a more immature phenotype were then selected by preferential fibronectin adhesion (FN-BMMSCs) for use in subsequent in vitro assays. DDM at 10 µg/mL reduced cell expansion, attenuated apoptosis and was the minimal concentration capable of inducing osteoblastic differentiation. Enzyme-linked immunosorbent assay (ELISA) quantification of growth factors indicated physiological levels produced the above responses; transforming growth factor β (TGF-β1) was predominant (15.6 ng/mg DDM), with relatively lower concentrations of BMP-2, FGF, VEGF and PDGF (6.2-4.7 ng/mg DDM). Fractionation of growth factors from other DDM components by heparin affinity chromatography diminished osteogenic responses. Depletion of biglycan from DDM also attenuated osteogenic potency, which was partially rescued by the isolated biglycan. Decorin depletion from DDM had no influence on osteogenic potency. Collectively, these results demonstrate the potential of DDM for the delivery of physiological levels of growth factors for bone repair processes, and substantiate a role for biglycan as an additional adjuvant for driving osteogenic pathways.

The available evidence on demineralised bone matrix in trauma and orthopaedic surgery

NARCIS (Netherlands)

J. van der Stok (Johan); Hartholt, K.A.; Schoenmakers, D.A.L.; J.J. Arts (Jacobus J.)

2017-01-01

textabstractObjectives: The aim of this systematic literature review was to assess the clinical level of evidence of commercially available demineralised bone matrix (DBM) products for their use in trauma and orthopaedic related surgery. Methods: A total of 17 DBM products were used as search terms

Molecular beacon-based real-time PCR method for detection of porcine DNA in gelatin and gelatin capsules.

Science.gov (United States)

Mohamad, Nurhidayatul Asma; Mustafa, Shuhaimi; Khairil Mokhtar, Nur Fadhilah; El Sheikha, Aly Farag

2018-03-05

The pharmaceutical industry has boosted gelatin consumption worldwide. This is supported by the availability of cost-effective gelatin production from porcine by-products. However, cross-contamination of gelatin materials, where porcine gelatin was unintentionally included in the other animal sources of gelatin, has caused significant concerns about halal authenticity. The real-time polymerase chain reaction (PCR) has enabled a highly specific and sensitive animal species detection method in various food products. Hence, such a technique was employed in the present study to detect and quantify porcine DNA in gelatin using a molecular beacon probe, with differences in performance between mitochondrial (cytochrome b gene) and chromosomal DNA-(MPRE42 repetitive element) based porcine-specific PCR assays being compared. A higher sensitivity was observed in chromosomal DNA (MPRE-PCR assay), where this assay allows the detection of gelatin DNA at amounts as as low as 1 pg, whereas mitochondrial DNA (CBH-PCR assay) can only detect at levels down to 10 pg of gelatin DNA. When an analysis with commercial gelatin and gelatin capsule samples was conducted, the same result was observed, with a significantly more sensitive detection being provided by the repetitive element of chromosomal DNA. The present study has established highly sensitive DNA-based porcine detection systems derived from chromosomal DNA that are feasible for highly processed products such as gelatin and gelatin capsules containing a minute amount of DNA. This sensitive detection method can also be implemented to assist the halal authentication process of various food products available on the market. © 2018 Society of Chemical Industry. © 2018 Society of Chemical Industry.

THE MEASUREMENT OF BONE QUALITY USING GRAY LEVEL CO-OCCURRENCE MATRIX TEXTURAL FEATURES.

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Shirvaikar, Mukul; Huang, Ning; Dong, Xuanliang Neil

2016-10-01

In this paper, statistical methods for the estimation of bone quality to predict the risk of fracture are reported. Bone mineral density and bone architecture properties are the main contributors of bone quality. Dual-energy X-ray Absorptiometry (DXA) is the traditional clinical measurement technique for bone mineral density, but does not include architectural information to enhance the prediction of bone fragility. Other modalities are not practical due to cost and access considerations. This study investigates statistical parameters based on the Gray Level Co-occurrence Matrix (GLCM) extracted from two-dimensional projection images and explores links with architectural properties and bone mechanics. Data analysis was conducted on Micro-CT images of 13 trabecular bones (with an in-plane spatial resolution of about 50μm). Ground truth data for bone volume fraction (BV/TV), bone strength and modulus were available based on complex 3D analysis and mechanical tests. Correlation between the statistical parameters and biomechanical test results was studied using regression analysis. The results showed Cluster-Shade was strongly correlated with the microarchitecture of the trabecular bone and related to mechanical properties. Once the principle thesis of utilizing second-order statistics is established, it can be extended to other modalities, providing cost and convenience advantages for patients and doctors.

Identification of transcriptional macromolecular associations in human bone using browser based in silico analysis in a giant correlation matrix.

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Reppe, Sjur; Sachse, Daniel; Olstad, Ole K; Gautvik, Vigdis T; Sanderson, Paul; Datta, Harish K; Berg, Jens P; Gautvik, Kaare M

2013-03-01

Intracellular signaling is critically dependent on gene regulatory networks comprising physical molecular interactions. Presently, there is a lack of comprehensive databases for most human tissue types to verify such macromolecular interactions. We present a user friendly browser which helps to identify functional macromolecular interactions in human bone as significant correlations at the transcriptional level. The molecular skeletal phenotype has been characterized by transcriptome analysis of iliac crest bone biopsies from 84 postmenopausal women through quantifications of ~23,000 mRNA species. When the signal levels were inter-correlated, an array containing >260 million correlations was generated, thus recognizing the human bone interactome at the RNA level. The matrix correlation and p values were made easily accessible by a freely available online browser. We show that significant correlations within the giant matrix are reproduced in a replica set of 13 male vertebral biopsies. The identified correlations differ somewhat from transcriptional interactions identified in cell culture experiments and transgenic mice, thus demonstrating that care should be taken in extrapolating such results to the in vivo situation in human bone. The current giant matrix and web browser are a valuable tool for easy access to the human bone transcriptome and molecular interactions represented as significant correlations at the RNA-level. The browser and matrix should be a valuable hypothesis generating tool for identification of regulatory mechanisms and serve as a library of transcript relationships in human bone, a relatively inaccessible tissue. Copyright © 2012 Elsevier Inc. All rights reserved.

Design and Fabrication of Biodegradable Porous Chitosan/Gelatin/Tricalcium Phosphate Hybrid Scaffolds for Tissue Engineering

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Y. Mohammadi

2007-08-01

Full Text Available In this study, based on a biomimetic approach, novel 3D biodegradable porous hybrid scaffolds consisting of chitosan, gelatin, and tricalcium phosphate were developed for bone and cartilage tissue engineering. Macroporous chitosan/ gelatin/β-TCP scaffolds were prepared through the process of freeze-gelation/solid-liquid phase separation. The results showed that the prepared scaffolds are highly porous, with porosities larger than 80%, and have interconnected pores. Biocompatibility studies were successfully performed by in vitro and in vivo assays. Moreover, the attachment, migration, and proliferation of chondrocytes on these unique temporary scaffolds were examined to determine their potentials in tissue engineering applications.

Enhanced Sternal Healing via Platelet-Rich Plasma and Biodegradable Gelatin Hydrogel.

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Shibata, Masafumi; Takagi, Gen; Kudo, Mitsuhiro; Kurita, Jiro; Kawamoto, Yoko; Miyagi, Yasuo; Kanazashi, Mikimoto; Sakatani, Takashi; Naito, Zenya; Tabata, Yasuhiko; Miyamoto, Masaaki; Nitta, Takashi

2018-05-16

Platelet-rich plasma (PRP) contains numerous growth factors and promotes bone fracture healing. The aim of this study was to evaluate the effectiveness of the controlled release of PRP from biodegradable gelatin hydrogel for promoting healing in a rabbit ischemic sternal model. PRP was prepared from the whole blood of a Japanese white rabbit. Sixteen rabbits were randomized into four groups (each n = 4) and all underwent median sternotomy and bilateral internal thoracic artery removal. Before the sternum was closed, the following solutions were applied between the sternum incisions in three of the groups: 30 mg of gelatin hydrogel incorporating 300 μL of phosphate-buffered saline, 300 μL of a solution form of PRP, or 30 mg of gelatin hydrogel incorporating 300 μL of PRP (PRP+Gel). The fourth group acted as a control. Sternal healing was evaluated by histology and micro-computed tomography 7 days after the intervention. The PRP+Gel group showed a significantly higher proportion of fibrosis within the fracture area (an indicator of sternal healing) than the other groups and a significantly higher mean intensity of osteocalcin. These results indicate that the controlled release of PRP from locally applied gelatin hydrogel was markedly effective in enhancing sternal healing in the early postoperative period. This novel therapy could potentially help prevent complications such as deep sternal wound infection and could result in early postoperative ambulation after median sternotomy.

Effect of platelet-rich plasma combined with demineralised bone matrix on bone healing in rabbit ulnar defects.

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Galanis, Vasilios; Fiska, Alice; Kapetanakis, Stylianos; Kazakos, Konstantinos; Demetriou, Thespis

2017-09-01

This study evaluates the effect of autologous platelet-rich plasma (PRP) combined with xenogeneic demineralised bone matrix (DBM) on bone healing of critical-size ulnar defects (2-2.5 times the ulnar diameter) in New Zealand White rabbits. Critical-size defects were created unilaterally in the ulna of 36 rabbits, while keeping the contralateral limb intact. They were divided into three groups. In Group A, the defect was filled with autologous PRP and in Group B, with autologous PRP combined with DBM; in Group C, the defect remained empty. The rabbits were euthanised 12 weeks postoperatively. Radiological, biomechanical and histological assessments were carried out and statistical analysis of the results was performed. Group B had significantly higher radiological and histological scores than Groups A and C. Defects in Group B showed significant new bone formation, whereas there was minimal or no new bone formation in Groups A and C. Only specimens in Group B showed macroscopic bone union. Biomechanical evaluation of the treated and intact contralateral limbs in Group B showed significant differences. In this study, statistically significant enhancement of bone healing was found in critical-size defects treated with PRP and DBM, as shown by radiological findings, gross assessment, and biomechanical and histopathological results. Defects in the two other groups remained unbridged. Therefore, PRP was effective only when it was used in combination with a bone graft. Copyright: © Singapore Medical Association

The Effects of Ribose on Mechanical and Physicochemical Properties of Cold Water Fish Gelatin Films

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Neda Javadian

2014-06-01

Full Text Available Native fish gelatin has some disadvantages such as high hydrophilic, and solubility in cold water. Mixing with other biopolymers and crosslinking by sugars may improve functional properties of fish gelatin. So in this research, the effects of ribose were investigated on moisture sorption isotherm, solubility in water, and mechanical properties of cold water fish gelatin (CWFG films. Ribose sugar was incorporated into CWFG solutions at different concentrations (e.g. 0, 2, 4, and 6% w/w dried gelatin. Physicochemical properties such as water solubility, moisture sorption isotherm and mechanical properties of the films were measured according to ASTM standards. Results showed that incorporation of ribose sugar significantly improved functional properties of CWFG films. Solubility, moisture content and monolayer water content of the matrixes were decreased by increasing the ribose contents. Mechanical properties of biocomposites were improved more than 20% and moisture sorption isotherm curve significantly shifted to lower moisture contents. The results of this study could be explored for commercial use, depending on industrial needs for either production of edible films or for packaging purposes.

Origin Determination and Differentiation of Gelatin Species of Bovine, Porcine, and Piscine through Analytical Methods

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Hatice Saadiye Eryılmaz

2017-06-01

Full Text Available Gelatin origin determination has been a crucial issue with respect to religion and health concerns. It is necessary to analyze the origin of gelatin with reliable methods to ensure not only consumer choices but also safety and legal requirements such as labeling. There are many analytical methods developed for detection and/or quantification of gelatin from different sources including bovine, porcine and piscine. These analytical methods can be divided into physicochemical, chromatographic, immunochemical, spectroscopic and molecular methods. Moreover, computational methods have been used in some cases consecutively to ensure sensitivity of the analytical methods. Every method has different advantages and limitations due to their own principles, applied food matrix and process conditions of material. The present review intends to give insight into novel analytical methods and perspectives that have been developed to differentiate porcine, bovine and piscine gelatins and to establish their authenticity. Almost every method can be succeeded in origin determination; however, it is a matter of sensitivity in that some researches fail to ensure sufficient differentiation.

Three-Dimensional Cone Beam Computed Tomography Volumetric Outcomes of rhBMP-2/Demineralized Bone Matrix versus Iliac Crest Bone Graft for Alveolar Cleft Reconstruction.

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Liang, Fan; Yen, Stephen L-K; Imahiyerobo, Thomas; Sanborn, Luke; Yen, Leia; Yen, Daniel; Nazarian, Sheila; Jedrzejewski, Breanna; Urata, Mark; Hammoudeh, Jeffrey

2017-10-01

Recent studies indicate that recombinant human bone morphogenetic protein-2 (rhBMP-2) in a demineralized bone matrix scaffold is a comparable alternative to iliac bone autograft in the setting of secondary alveolar cleft repair. Postreconstruction occlusal radiographs demonstrate improved bone stock when rhBMP-2/demineralized bone matrix (DBM) scaffold is used but lack the capacity to evaluate bone growth in three dimensions. This study uses cone beam computed tomography to provide the first clinical evaluation of volumetric and density comparisons between these two treatment modalities. A prospective study was conducted with 31 patients and 36 repairs of the alveolar cleft over a 2-year period. Twenty-one repairs used rhBMP-2/DBM scaffold and 14 repairs used iliac bone grafting. Postoperatively, occlusal radiographs were obtained at 3 months to evaluate bone fill; cone beam computed tomographic images were obtained at 6 to 9 months to compare volumetric and density data. At 3 months, postoperative occlusal radiographs demonstrated that 67 percent of patients receiving rhBMP-2/DBM scaffold had complete bone fill of the alveolus, versus 56 percent of patients in the autologous group. In contrast, cone beam computed tomographic data showed 31.6 percent (95 percent CI, 24.2 to 38.5 percent) fill in the rhBMP-2 group compared with 32.5 percent (95 percent CI, 22.1 to 42.9 percent) in the autologous population. Density analysis demonstrated identical average values between the groups (1.38 g/cc). These data demonstrate comparable bone regrowth and density values following secondary alveolar cleft repair using rhBMP-2/DBM scaffold versus autologous iliac bone graft. Cone beam computed tomography provides a more nuanced understanding of true bone regeneration within the alveolar cleft that may contribute to the information provided by occlusal radiographs alone. Therapeutic, II.

Localized ridge defect augmentation using human pericardium membrane and demineralized bone matrix.

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Vidyadharan, Arun Kumar; Ravindran, Anjana

2014-01-01

Patient wanted to restore her lost teeth with implants in the lower left first molar and second premolar region. Cone beam computerized tomography (CBCT) revealed inadequate bone width and height around future implant sites. The extraction socket of second premolar area revealed inadequate socket healing with sparse bone fill after 4 months of extraction. To evaluate the clinical feasibility of using a collagen physical resorbable barrier made of human pericardium (HP) to augment localized alveolar ridge defects for the subsequent placement of dental implants. Ridge augmentation was done in the compromised area using Puros® demineralized bone matrix (DBM) Putty with chips and an HP allograft membrane. Horizontal (width) and vertical hard tissue measurements with CBCT were recorded on the day of ridge augmentation surgery, 4 month and 7 months follow-up. Intra oral periapical taken 1 year after implant installation showed minimal crestal bone loss. Bone volume achieved through guided bone regeneration was a gain of 4.8 mm horizontally (width) and 6.8 mm vertically in the deficient ridge within a period of 7 months following the procedure. The results suggested that HP Allograft membrane may be a suitable component for augmentation of localized alveolar ridge defects in conjunction with DBM with bone chips.

Alterations in proteins of bone marrow extracellular matrix in undernourished mice

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C.L. Vituri

2000-08-01

Full Text Available The objective of the present study was to determine the effect of protein malnutrition on the glycoprotein content of bone marrow extracellular matrix (ECM. Two-month-old male Swiss mice were submitted to protein malnutrition with a low-protein diet containing 4% casein as compared to 20% casein in the control diet. When the experimental group had attained a 20% loss of their original body weight, we extracted the ECM proteins from bone marrow with PBS buffer, and analyzed ECM samples by SDS-PAGE (7.5% and ECL Western blotting. Quantitative differences were observed between control and experimental groups. Bone marrow ECM from undernourished mice had greater amounts of extractable fibronectin (1.6-fold increase and laminin (4.8-fold increase when compared to the control group. These results suggest an association between fluctuations in the composition of the hematopoietic microenvironment and altered hematopoiesis observed in undernourished mice.

A composite demineralized bone matrix--self assembling peptide scaffold for enhancing cell and growth factor activity in bone marrow.

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Hou, Tianyong; Li, Zhiqiang; Luo, Fei; Xie, Zhao; Wu, Xuehui; Xing, Junchao; Dong, Shiwu; Xu, Jianzhong

2014-07-01

The need for suitable bone grafts is high; however, there are limitations to all current graft sources, such as limited availability, the invasive harvest procedure, insufficient osteoinductive properties, poor biocompatibility, ethical problems, and degradation properties. The lack of osteoinductive properties is a common problem. As an allogenic bone graft, demineralized bone matrix (DBM) can overcome issues such as limited sources and comorbidities caused by invasive harvest; however, DBM is not sufficiently osteoinductive. Bone marrow has been known to magnify osteoinductive components for bone reconstruction because it contains osteogenic cells and factors. Mesenchymal stem cells (MSCs) derived from bone marrow are the gold standard for cell seeding in tissue-engineered biomaterials for bone repair, and these cells have demonstrated beneficial effects. However, the associated high cost and the complicated procedures limit the use of tissue-engineered bone constructs. To easily enrich more osteogenic cells and factors to DBM by selective cell retention technology, DBM is modified by a nanoscale self-assembling peptide (SAP) to form a composite DBM/SAP scaffold. By decreasing the pore size and increasing the charge interaction, DBM/SAP scaffolds possess a much higher enriching yield for osteogenic cells and factors compared with DBM alone scaffolds. At the same time, SAP can build a cellular microenvironment for cell adhesion, proliferation, and differentiation that promotes bone reconstruction. As a result, a suitable bone graft fabricated by DBM/SAP scaffolds and bone marrow represents a new strategy and product for bone transplantation in the clinic. Copyright © 2014 Elsevier Ltd. All rights reserved.

Novel biocompatible polymeric blends for bone regeneration: Material and matrix design and development

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Deng, Meng

The first part of the work presented in this dissertation is focused on the design and development of novel miscible and biocompatible polyphosphazene-polyester blends as candidate materials for scaffold-based bone tissue engineering applications. Biodegradable polyesters such as poly(lactide-co-glycolide) (PLAGA) are among the most widely used polymeric materials for bone tissue engineering. However, acidic degradation products resulting from the bulk degradation mechanism often lead to catastrophic failure of the structure integrity, and adversely affect biocompatibility both in vitro and in vivo. One promising approach to circumvent these limitations is to blend PLAGA with other macromolecules that can buffer the acidic degradation products with a controlled degradation rate. Biodegradable polyphosphazenes (PPHOS), a new class of biomedical materials, have proved to be superior candidate materials to achieve this objective due to their unique buffering degradation products. A highly practical blending approach was adopted to develop novel biocompatible, miscible blends of these two polymers. In order to achieve this miscibility, a series of amino acid ester, alkoxy, aryloxy, and dipeptide substituted PPHOS were synthesized to promote hydrogen bonding interactions with PLAGA. Five mixed-substituent PPHOS compositions were designed and blended with PLAGA at different weight ratios producing candidate blends via a mutual solvent method. Preliminary characterization identified two specific side groups namely glycylglycine dipeptide and phenylphenoxy that resulted in improved blend miscibility and enhanced in vitro osteocompatibility. These findings led to the synthesis of a mixed-substituent polyphosphazene poly[(glycine ethyl glycinato)1(phenylphenoxy)1phosphazene] (PNGEGPhPh) for blending with PLAGA. Two dipeptide-based blends having weight ratios of PNGEGPhPh to PLAGA namely 25:75 (Matrix1) and 50:50 (Matrix2) were fabricated. Both of the blends were

Reconstitution of bone-like matrix in osteogenically differentiated mesenchymal stem cell–collagen constructs: A three-dimensional in vitro model to study hematopoietic stem cell niche

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WY Lai

2013-10-01

Full Text Available Mesenchymal stem/stromal cells (MSCs and osteoblasts are important niche cells for hematopoietic stem cells (HSCs in bone marrow osteoblastic niche. Here, we aim to partially reconstitute the bone marrow HSC niche in vitro using collagen microencapsulation for investigation of the interactions between HSCs and MSCs. Mouse MSCs (mMSCs microencapsulated in collagen were osteogenically differentiated to derive a bone-like matrix consisting of osteocalcin, osteopontin, and calcium deposits and secreted bone morphogenic protein 2 (BMP2. Decellularized bone-like matrix was seeded with fluorescence-labeled human MSCs and HSCs. Comparing with pure collagen scaffold, significantly more HSCs and HSC–MSC pairs per unit area were found in the decellularized bone-like matrix. Moreover, incubation with excess neutralizing antibody of BMP2 resulted in a significantly higher number of HSC per unit area than that without in the decellularized matrix. This work suggests that the osteogenic differentiated MSC–collagen microsphere is a valuable three-dimensional in vitro model to elucidate cell–cell and cell–matrix interactions in HSC niche.

An NMR relaxometry and gravimetric study of gelatin-free aqueous polyacrylamide dosimeters

International Nuclear Information System (INIS)

Babic, Steven; Schreiner, L John

2006-01-01

In conformal radiation therapy, a high dose of radiation is given to a target volume to increase the probability of cure, and care is taken to minimize the dose to surrounding healthy tissue. The techniques used to achieve this are very complicated and the precise verification of the resulting three-dimensional (3D) dose distribution is required. Polyacrylamide gelatin (PAG) dosimeters with magnetic resonance imaging and optical computed tomography scanning provide the required 3D dosimetry with high spatial resolution. Many basic studies have characterized these chemical dosimeters that polymerize under irradiation. However, the investigation of the fundamental properties of the radiation-induced polymerization in PAG dosimeters is complicated by the presence of the background gelatin matrix. In this work, a gelatin-free model system for the study of the basic radiation-induced polymerization in PAG dosimeters has been developed. Experiments were performed on gelatin-free dosimeters, named aqueous polyacrylamide (APA) dosimeters, containing equal amounts of acrylamide and N,N'-methylene-bisacrylamide. The APA dosimeters were prepared with four different total monomer concentrations (2, 4, 6 and 8% by weight). Nuclear magnetic resonance (NMR) spin-spin and spin-lattice proton relaxation measurements at 20 MHz, and gravimetric analyses performed on all four dosimeters, show a continuous degree of polymerization over the dose range of 0-25 Gy. The developed NMR model explains the relationship observed between the relaxation data and the amount of crosslinked polymer formed at each dose. This model can be extended with gelatin relaxation data to provide a fundamental understanding of radiation-induced polymerization in the conventional PAG dosimeters

Silk fibroin/gelatin-chondroitin sulfate-hyaluronic acid effectively enhances in vitro chondrogenesis of bone marrow mesenchymal stem cells.

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Sawatjui, Nopporn; Damrongrungruang, Teerasak; Leeanansaksiri, Wilairat; Jearanaikoon, Patcharee; Hongeng, Suradej; Limpaiboon, Temduang

2015-01-01

Tissue engineering is becoming promising for cartilage repair due to the limited self-repair capacity of cartilage tissue. We previously fabricated and characterized a three-dimensional silk fibroin/gelatin-chondroitin sulfate-hyaluronic acid (SF-GCH) scaffold and showed that it could promote proliferation of human bone marrow mesenchymal stem cells (BM-MSCs). This study aimed to evaluate its biological performance as a new biomimetic material for chondrogenic induction of BM-MSCs in comparison to an SF scaffold and conventional pellet culture. We found that the SF-GCH scaffold significantly enhanced the proliferation and chondrogenic differentiation of BM-MSCs compared to the SF scaffold and pellet culture in which the production of sulfated glycoaminoglycan was increased in concordance with the up-regulation of chondrogenic-specific gene markers. Our findings indicate the significant role of SF-GCH by providing a supportive structure and the mimetic cartilage environment for chondrogenesis which enables cartilage regeneration. Thus, our fabricated SF-GCH scaffold may serve as a potential biomimetic material for cartilage tissue engineering. Copyright © 2015 Elsevier B.V. All rights reserved.

Electric field-assisted formation of organically modified hydroxyapatite (ormoHAP) spheres in carboxymethylated gelatin gels.

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Heinemann, C; Heinemann, S; Kruppke, B; Worch, H; Thomas, J; Wiesmann, H P; Hanke, T

2016-10-15

A biomimetic strategy was developed in order to prepare organically modified hydroxyapatite (ormoHAP) with spherical shape. The technical approach is based on electric field-assisted migration of calcium ions and phosphate ions into a hydrogel composed of carboxymethylated gelatin. The electric field as well as the carboxymethylation using glucuronic acid (GlcA) significantly accelerates the mineralization process, which makes the process feasible for lab scale production of ormoHAP spheres and probably beyond. A further process was developed for gentle separation of the ormoHAP spheres from the gelatin gel without compromising the morphology of the mineral. The term ormoHAP was chosen since morphological analyses using electron microscopy (SEM, TEM) and element analysis (EDX, FT-IR, XRD) confirmed that carboxymethylated gelatin molecules use to act as organic templates for the formation of nanocrystalline HAP. The hydroxyapatite (HAP) crystals self-organize to form hollow spheres with diameters ranging from 100 to 500nm. The combination of the biocompatible chemical composition and the unique structure of the nanocomposites is considered to be a useful basis for future applications in functionalized degradable biomaterials. A novel bioinspired mineralization process was developed based on electric field-assisted migration of calcium and phosphate ions into biochemically carboxymethylated gelatin acting as organic template. Advantages over conventional hydroxyapatite include particle size distribution and homogeneity as well as achievable mechanical properties of relevant composites. Moreover, specifically developed calcium ion or phosphate ion release during degradation can be useful to adjust the fate of bone cells in order to manipulate remodeling processes. The hollow structure of the spheres can be useful for embedding drugs in the core, encapsulated by the highly mineralized outer shell. In this way, controlled drug release could be achieved, which enables

Halal authenticity of gelatin using species-specific PCR.

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Shabani, Hessam; Mehdizadeh, Mehrangiz; Mousavi, Seyed Mohammad; Dezfouli, Ehsan Ansari; Solgi, Tara; Khodaverdi, Mahdi; Rabiei, Maryam; Rastegar, Hossein; Alebouyeh, Mahmoud

2015-10-01

Consumption of food products derived from porcine sources is strictly prohibited in Islam. Gelatin, mostly derived from bovine and porcine sources, has many applications in the food and pharmaceutical industries. To ensure that food products comply with halal regulations, development of valid and reliable analytical methods is very much required. In this study, a species-specific polymerase chain reaction (PCR) assay using conserved regions of mitochondrial DNA (cytochrome b gene) was performed to evaluate the halal authenticity of gelatin. After isolation of DNA from gelatin powders with known origin, conventional PCR using species-specific primers was carried out on the extracted DNA. The amplified expected PCR products of 212 and 271 bp were observed for porcine and bovine gelatin, respectively. The sensitivity of the method was tested on binary gelatin mixtures containing 0.1%, 1%, 10%, and 100% (w/w) of porcine gelatin within bovine gelatin and vice versa. Although most of the DNA is degraded due to the severe processing steps of gelatin production, the minimum level of 0.1% w/w of both porcine and bovine gelatin was detected. Moreover, eight food products labeled as containing bovine gelatin and eight capsule shells were subjected to PCR examination. The results showed that all samples contained bovine gelatin, and the absence of porcine gelatin was verified. This method of species authenticity is very useful to verify whether gelatin and gelatin-containing food products are derived from halal ingredients. Copyright © 2015 Elsevier Ltd. All rights reserved.

On spray drying of oxidized corn starch cross-linked gelatin microcapsules for drug release.

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Dang, Xugang; Yang, Mao; Shan, Zhihua; Mansouri, Shahnaz; May, Bee K; Chen, Xiaodong; Chen, Hui; Woo, Meng Wai

2017-05-01

Spray-dried gelatin/oxidized corn starch (G/OCS) microcapsules were produced for drug release application. The prepared microcapsules were characterized through a scanning electron microscope (SEM) picture and thermogravimetric analysis (TGA). The swelling characteristics of the G/OCS microcapsules and release properties of vitamin C were then investigated. The results from structural analysis indicated that the presence of miscibility and compatibility between oxidized corn starch and gelatin, and exhibits high thermal stability up to 326°C. The swelling of G/OCS microcapsules increased with increasing pH and reduced with decreasing ionic strength, attributed to the cross-linking between gelatin and oxidized corn starch, ionization of functional groups. Vitamin C release characteristic revealed controlled release behavior in the first 3h of contact with an aqueous medium. This release behavior was independent of the swelling behavior indicating the potential of the encapsulating matrix to produce controlled release across a spectrum of pH environment. Copyright © 2016 Elsevier B.V. All rights reserved.

Fabrication and characterisation of biomimetic, electrospun gelatin fibre scaffolds for tunica media-equivalent, tissue engineered vascular grafts

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Elsayed, Y. [Advanced Materials Group, University of Surrey, Guildford, Surrey GU2 7XH (United Kingdom); Lekakou, C., E-mail: C.Lekakou@surrey.ac.uk [Advanced Materials Group, University of Surrey, Guildford, Surrey GU2 7XH (United Kingdom); Labeed, F. [Centre of Biomedical Engineering, University of Surrey, Guildford, Surrey GU2 7XH (United Kingdom); Tomlins, P. [National Physical Laboratory (NPL), Teddington, Middlesex TW11 0LW (United Kingdom)

2016-04-01

It is increasingly recognised that biomimetic, natural polymers mimicking the extracellular matrix (ECM) have low thrombogenicity and functional motifs that regulate cell–matrix interactions, with these factors being critical for tissue engineered vascular grafts especially grafts of small diameter. Gelatin constitutes a low cost substitute of soluble collagen but gelatin scaffolds so far have shown generally low strength and suture retention strength. In this study, we have devised the fabrication of novel, electrospun, multilayer, gelatin fibre scaffolds, with controlled fibre layer orientation, and optimised gelatin crosslinking to achieve not only compliance equivalent to that of coronary artery but also for the first time strength of the wet tubular acellular scaffold (swollen with absorbed water) same as that of the tunica media of coronary artery in both circumferential and axial directions. Most importantly, for the first time for natural scaffolds and in particular gelatin, high suture retention strength was achieved in the range of 1.8–1.94 N for wet acellular scaffolds, same or better than that for fresh saphenous vein. The study presents the investigations to relate the electrospinning process parameters to the microstructural parameters of the scaffold, which are further related to the mechanical performance data of wet, crosslinked, electrospun scaffolds in both circumferential and axial tubular directions. The scaffolds exhibited excellent performance in human smooth muscle cell (SMC) proliferation, with SMCs seeded on the top surface adhering, elongating and aligning along the local fibres, migrating through the scaffold thickness and populating a transverse distance of 186 μm and 240 μm 9 days post-seeding for scaffolds of initial dry porosity of 74 and 83%, respectively. - Highlights: • Novel crosslinked electrospun gelatin scaffolds of specific fibre layer orientation • These scaffolds have compliance equivalent to that of coronary

A Novel High Mechanical Property PLGA Composite Matrix Loaded with Nanodiamond-Phospholipid Compound for Bone Tissue Engineering.

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Zhang, Fan; Song, Qingxin; Huang, Xuan; Li, Fengning; Wang, Kun; Tang, Yixing; Hou, Canglong; Shen, Hongxing

2016-01-20

A potential bone tissue engineering material was produced from a biodegradable polymer, poly(lactic-co-glycolic acid) (PLGA), loaded with nanodiamond phospholipid compound (NDPC) via physical mixing. On the basis of hydrophobic effects and physical absorption, we modified the original hydrophilic surface of the nanodiamond (NDs) with phospholipids to be amphipathic, forming a typical core-shell structure. The ND-phospholipid weight ratio was optimized to generate sample NDPC50 (i.e., ND-phospholipid weight ratio of 100:50), and NDPC50 was able to be dispersed in a PLGA matrix at up to 20 wt %. Compared to a pure PLGA matrix, the introduction of 10 wt % of NDPC (i.e., sample NDPC50-PF10) resulted in a significant improvement in the material's mechanical and surface properties, including a decrease in the water contact angle from 80 to 55°, an approximately 100% increase in the Young's modulus, and an approximate 550% increase in hardness, thus closely resembling that of human cortical bone. As a novel matrix supporting human osteoblast (hFOB1.19) growth, NDPC50-PFs with different amounts of NDPC50 demonstrated no negative effects on cell proliferation and osteogenic differentiation. Furthermore, we focused on the behaviors of NDPC-PFs implanted into mice for 8 weeks and found that NDPC-PFs induced acceptable immune response and can reduce the rapid biodegradation of PLGA matrix. Our results represent the first in vivo research on ND (or NDPC) as nanofillers in a polymer matrix for bone tissue engineering. The high mechanical properties, good in vitro and in vivo biocompatibility, and increased mineralization capability suggest that biodegradable PLGA composite matrices loaded with NDPC may potentially be useful for a variety of biomedical applications, especially bone tissue engineering.

Three-dimensional CaP/gelatin lattice scaffolds with integrated osteoinductive surface topographies for bone tissue engineering

International Nuclear Information System (INIS)

Nadeem, Danish; Su, Bo; Smith, Carol-Anne; Dalby, Matthew J; Dominic Meek, R M; Lin, Sien; Li, Gang

2015-01-01

Surface topography is known to influence stem cells and has been widely used as physical stimuli to modulate cellular behaviour including adhesion, proliferation and differentiation on 2D surfaces. Integration of well-defined surface topography into three-dimensional (3D) scaffolds for tissue engineering would be useful to direct the cell fate for intended applications. Technical challenges are remaining as how to fabricate such 3D scaffolds with controlled surface topography from a range of biodegradable and biocompatible materials. In this paper, a novel fabrication process using computer numerically controlled machining and lamination is reported to make 3D calcium phosphate/gelatin composite scaffolds with integrated surface micropatterns that are introduced by embossing prior to machining. Geometric analysis shows that this method is versatile and can be used to make a wide range of lattices with porosities that meet the basic requirements for bone tissue engineering. Both in vitro and in vivo studies show that micropatterned composite scaffolds with surfaces comprising 40 μm pits and 50 μm grooves were optimal for improved osteogenesis. The results have demonstrated the potential of a novel fabrication process for producing cell-instructive scaffolds with designed surface topographies to induce specific tissue regeneration. (paper)

Changes in subchondral bone mineral density and collagen matrix organization in growing horses.

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Holopainen, Jaakko T; Brama, Pieter A J; Halmesmäki, Esa; Harjula, Terhi; Tuukkanen, Juha; van Weeren, P René; Helminen, Heikki J; Hyttinen, Mika M

2008-12-01

The effects of growth and maturation on the mineral deposition and the collagen framework of equine subchondral bone (SCB) were studied. Osteochondral specimens (diameter 6 mm) from the left metacarpophalangeal joint of 5-(n=8), 11-(n=8) and 18-month-old (n=6) horses were investigated at two differently loaded sites (Site 1 (S1): intermittent peak loading; Site 2 (S2): habitual loading). The SCB mineral density (BMD) was measured with peripheral quantitative computer tomography (pQCT), and the data were adjusted against the volume fraction (Vv) of the bone extracellular matrix (ECM). Polarised light microscopy (PLM) was used to analyze the Vv, the collagen fibril parallelism index and the orientation angle distribution in two fractions (1 mm/fraction) beneath the osteochondral junction of the SCB. PLM analysis was made along two randomly selected perpendicularly oriented vertical sections to measure the tissue anisotropy in the x-, y-, and z-directions. The BMD of SCB at S1 and S2 increased significantly during maturation. At the same time, the Vv of the ECM increased even more. This meant that the Vv-adjusted BMD decreased. There were no significant differences between sites. The basic collagen fibril framework of SCB seems to be established already at the age of 5 months. During maturation, the extracellular matrix underwent a decrease in collagen fibril parallelism but no changes in collagen orientation. The variation was negligible in the collagen network estimates in the two section planes. Growth and maturation induce significant changes in the equine SCB. The BMD increase in SCB is primarily due to the growth of bone volume and not to any increase in mineral deposition. An increase in weight-bearing appears to greatly affect the BMD and the volume of the extracellular matrix. Growth and maturation induce a striking change in collagen fibril parallelism but not in fibril orientation. The structural anisotropy of the subchondral bone is significant along the

Effect of electron beam irradiation on mechanical properties of gelatin/Brazil nut shell fiber composites

International Nuclear Information System (INIS)

Inamura, Patricia Y.; Shimazaki, Kleber; Moura, Esperidiana Augusta Barretos de; Mastro, Nelida L. del; Colombo, Maria Aparecida; Rosa, Ricardo de

2010-01-01

The use of natural fiber as polymeric matrix reinforcement has attracted interest, as fibers are renewable, of low cost, biodegradable and possesses non-toxic properties. In the present paper, Brazil nuts (Bertholletia excelsa) shell fiber (10% w/w) were mixed with gelatin (25% w/w), glycerin as plasticizer and acrylamide as copolymer to investigate the resultant mechanical properties effects upon ionizing radiation. The samples were irradiated at 40 kGy using a Dynamitron electron beam accelerator, at room temperature in the presence of air. The results showed that samples of gelatin with 10% of Brazil nuts shell fiber and irradiated at 40 kGy presented promising results for mechanical performance. (author)

Detection of porcine DNA in gelatine and gelatine-containing processed food products-Halal/Kosher authentication.

Science.gov (United States)

Demirhan, Yasemin; Ulca, Pelin; Senyuva, Hamide Z

2012-03-01

A commercially available real-time PCR, based on a multi-copy target cytochrome b (cyt b) using porcine specific primers, has been validated for the Halal/Kosher authentication of gelatine. Extraction and purification of DNA from gelatine were successfully achieved using the SureFood® PREP Animal system, and real-time PCR was carried out using SureFood® Animal ID Pork Sens kit. The minimum level of adulteration that could be detected was 1.0% w/w for marshmallows and gum drops. A small survey was undertaken of processed food products such as gum drops, marshmallows and Turkish delight, believed to contain gelatine. Of fourteen food products from Germany, two samples were found to contain porcine gelatine, whereas of twenty-nine samples from Turkey twenty-eight were negative. However, one product from Turkey contained porcine DNA and thus was not Halal, and neither was the use of porcine gelatine indicated on the product label. Copyright © 2011 Elsevier Ltd. All rights reserved.

Ridge augmentation using recombinant human fibroblast growth factor-2 with biodegradable gelatin sponges incorporating β-tricalcium phosphate: a preclinical study in dogs.

Science.gov (United States)

Hoshi, S; Akizuki, T; Matsuura, T; Ikawa, T; Kinoshita, A; Oda, S; Tabata, Y; Matsui, M; Izumi, Y

2016-02-01

Fibroblast growth factor-2 (FGF-2) regulates the proliferation and differentiation of osteogenic cells, resulting in the promotion of bone formation. Biodegradable gelatin sponges incorporating β-tricalcium phosphate (β-TCP) have been reported as a scaffold, which has the ability to control growth factor release, offering sufficient mechanical strength and efficient migration of mesenchymal cells. In this study, we evaluated the effects of the combined use of recombinant human FGF-2 (rhFGF-2) and gelatin/β-TCP sponge on ridge augmentation in dogs. Six male beagle dogs were used in this study. Twelve wk after tooth extraction, bilateral 10 × 5 mm (width × depth) saddle-type defects were created 3 mm apart from the mesial side of the maxillary canine. At the experimental sites, the defects were filled with gelatin/β-TCP sponge infiltrated with 0.3% rhFGF-2, whereas gelatin/β-TCP sponge infiltrated with saline was applied to the control sites. Eight wk after surgery, qualitative and quantitative analyses were performed. There were no signs of clinical inflammation at 8 wk after surgery. Histometric measurements revealed that new bone height at the experimental sites (2.98 ± 0.65 mm) was significantly greater than that at the control sites (1.56 ± 0.66 mm; p = 0.004). The total tissue height was greater at the experimental sites (6.62 ± 0.66 mm) than that at the control sites (5.95 ± 0.74 mm), although there was no statistical significant difference (p = 0.051). Cast model measurements revealed that the residual defect height at the experimental sites (2.31 ± 0.50 mm) was significantly smaller than that at the control sites (3.51 ± 0.78 mm; p = 0.012). The combined use of rhFGF-2 and gelatin/β-TCP sponge promotes ridge augmentation in canine saddle-type bone defects. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Semisolid matrix-filled hard gelatin capsules for rapid dissolution of amlodipine besilate: Development and assessment

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Vijay K Tyagi

2013-01-01

Full Text Available The objective of the study was to prepare semisolid capsules (SSCs of poorly water-soluble drug amlodipine besilate (AB using a combination of technologies involving solid dispersion (SD preparation and converting it into semisolid matrix filled in hard gelatin capsules (termed as SSCs with the aim of reducing lag time in drug release and to improve the dissolution rate. AB is used for its anti-arrhythmic, anti-anginal, and anti-hypertensive activity. These are the emergency activities which should be treated as fast as possible like in the case of angina attack (heart attack. Any lag time that is generated due to its poor dissolution can add on in this emergency and that can be avoided by developing a readily dissolvable formulation: SDs of AB. SD of AB was prepared by fusion method using varying combinations of Poloxamer 407 and Plasdone S630. A total of nine batches (SD1−SD9 were characterized for the in vitro dissolution behavior in phosphate buffer pH7.4. SD8 with 95.8% cumulative drug release in 60 min, t50% = 4.1 min and DE 30 Min = 84.2% were selected for the development of the semisolid matrix. Differential scanning calorimetry of SD8 revealed molecular dispersion of AB and Plasdone S630 in Poloxamer 407. SD8 was then formulated as SSCs using gelucire 44/14 and PEG 400 as semisolid components and PEG 6000 as a suspending agent to achieve the reduction in lag time for drug release. A total of seven SSC formulations were prepared and evaluated for drug release. Formulation of SSC4 showed maximum cumulative drug release (CDR of 98.9% within 20 min that was almost a threefold reduction in the time required to achieve similar CDR by SD of AB. Thus, SSCs present an excellent approach to enhance the dissolution as well as to reduce the lag time of dissolution for poor water-soluble drugs especially to those therapeutic classes that are intended for faster onset of action.

Physicochemical and osteoplastic characteristics of 3D printed bone grafts based on synthetic calcium phosphates and natural polymers

Science.gov (United States)

Nezhurina, E. K.; Karalkin, P. A.; Komlev, V. S.; Sviridova, I. K.; Kirsanova, V. A.; Akhmedova, S. A.; Shanskiy, Ya D.; Fedotov, A. Yu; Barinov, S. M.; Sergeeva, N. S.

2018-04-01

A creation of personalized implants for regeneration of bone tissue seems to be a very promising biomedical technological approach. We have studied the physicochemical characteristics, cyto- and biocompatibility of three-dimensional constructs based on sodium alginate and gelatin in combination with 2 types of calcium phosphate (tricalcium phosphate or octacalcium phosphate) obtained by inkjet 3D printing. In our experiments, we have studied the physical and chemical properties of the constructs – their porosity, chemical composition, microarchitecture of the surface and mechanical elasticity. The cytocompatibility of 3D constructs and matrix-for-cell properties were investigated in vitro on a model of human osteosarcoma MG-63 cell line by means of MTT assay. The biocompatibility of 3D constructs was studied on the model of subcutaneous implantation in mice up to 12 weeks. All types of 3D constructs were cytocompatible in vitro, demonstrated good matrix-for-cells properties, and had supported cell proliferation for 2 weeks. In results of subcutaneous in vivo test all constructs demonstrated biocompatibility with slow bioresorption of organic and inorganic components. Osteogenesis proceeded more actively in rat tibia model defects (marginal excision), substituted by 3D printed 3-component implants based on alginate, gelatin and octacalcium phosphate.

Electrospun Zein/Gelatin Scaffold-Enhanced Cell Attachment and Growth of Human Periodontal Ligament Stem Cells

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Fanqiao Yang

2017-10-01

Full Text Available Periodontitis is a widespread dental disease affecting 10 to 15% of worldwide adult population, yet the current treatments are far from satisfactory. The human periodontal ligament stem cell is a promising potential seed cell population type in cell-based therapy and tissue regeneration, which require appropriate scaffold to provide a mimic extracellular matrix. Zein, a native protein derived from corn, has an excellent biodegradability, and therefore becomes a hotspot on research and application in the field of biomaterials. However, the high hydrophobicity of zein is unfavorable for cell adhesion and thus greatly limits its use. In this study, we fabricate co-electrospun zein/gelatin fiber scaffolds in order to take full advantages of the two natural materials and electrospun fiber structure. Zein and gelatin in four groups of different mass ratios (100:00, 100:20, 100:34, 100:50, and dissolved the mixtures in 1,1,1,3,3,3-hexafluoro-2-propanol, then produced membranes by electrospinning. The results showed that the scaffolds were smooth and homogeneous, as shown in scanning electron micrographs. The diameter of hybrid fibers was increased from 69 ± 22 nm to 950 ± 356 nm, with the proportion of gelatin increase. The cell affinity of zein/gelatin nanofibers was evaluated by using human periodontal ligament stem cells. The data showed that hydrophilicity and cytocompatibility of zein nanofibers were improved by blended gelatin. Taken together, our results indicated that the zein/gelatin co-electrospun fibers had sufficient mechanical properties, satisfied cytocompatibility, and can be utilized as biological scaffolds in the field of tissue regeneration.

Identification of osteocalcin as a permanent aging constituent of the bone matrix: basis for an accurate age at death determination.

Science.gov (United States)

Ritz, S; Turzynski, A; Schütz, H W; Hollmann, A; Rochholz, G

1996-01-12

Age at death determination based on aspartic acid racemization in dentin has been applied successfully in forensic odontology for several years now. An age-dependent accumulation of D-aspartic acid has also recently been demonstrated in bone osteocalcin, one of the most abundant noncollagenous proteins of the organic bone matrix. Evaluation of these initial data on in vivo racemization of aspartic acid in bone osteocalcin was taken a step further. After purification of osteocalcin from 53 skull bone specimens, the extent of aspartic acid racemization in this peptide was determined. The D-aspartic acid content of purified bone osteocalcin exhibited a very close relationship to age at death. This confirmed identification of bone osteocalcin as a permanent, 'aging' peptide of the organic bone matrix. Its D-aspartic acid content may be used as a measure of its age and hence that of the entire organism. The new biochemical approach to determination of age at death by analyzing bone is complex and demanding from a methodologic point of view, but appears to be superior in precision and reproducibility to most other methods applicable to bone.

Matrix metalloproteinases (MMP-2 and MMP-9) activity in corneal ulcer and ocular surface disorders determined by gelatin zymography.

Science.gov (United States)

Singh, Arti; Maurya, O P S; Jagannadhan, M V; Patel, Ashok

2012-01-01

The purpose of this paper is to determine the active form of matrix metalloproteinases (MMP-2 and MMP-9) in corneal ulcer and ocular surface disorder patients. A total of 35 patients of corneal ulcer, 20 patients of ocular surface disorders and 10 control subjects were included in this study and estimation of active form of MMP-2 and MMP-9 was done by gelatin zymography. Tear samples were collected by capillary tube method. Both pro- and active forms of MMP-9 were detected in 24 out of 35 patients with corneal ulcer and 15 out of 20 patients with ocular surface disorders. None of the patients were showing MMP-2 activity. Neither MMP-2 nor MMP-9 was detected in the control group. Active forms of MMP-9 are present in tears of severe ulcerative and ocular surface disorder patients. Thus, proteinase inhibitors have been recommended for the treatment of corneal ulcer and ocular surface disorders to reduced the progression of stromal ulcer and to minimize corneal scarring.

Studies on electronic structure of interfaces between Ag and gelatin for stabilization of Ag nanoparticles

International Nuclear Information System (INIS)

Tani, Tadaaki; Uchida, Takayuki

2015-01-01

Extremely high stability of Ag nanoparticles in photographic materials has forced us to study the electronic structures of the interfaces between thin layers of Ag, Au, and Pt and their surface membranes in ambient atmosphere by photoelectron yield spectroscopy in air and Kelvin probe method. Owing to the Fermi level equalization between a metal layer and a membrane coming from air, the electron transfer took place from the membrane to Pt and Au layers and from an Ag layer to the membrane, giving the reason for poor stability of Ag nanoparticles in air. The control of the Fermi level of an Ag layer with respect to that of a gelatin membrane in air could be widely made according to Nernst's equation by changing the pH and pAg values of an aqueous gelatin solution used to form the membrane, and thus available to stabilize Ag nanoparticles in a gelatin matrix. (author)

Biochemical studies of the macromolecular matrix of long bones in the Op/Orl mutant rat strain

Energy Technology Data Exchange (ETDEWEB)

Moczar, E; Berenholc, S; Phan-Dinh-Tuy, B; Robert, A M

1978-01-01

The long bones of normal and Op/Orl mutant rats were incubated with /sup 14/C-glucose and fractionated by EDTA and urea extraction. The analytical results of the various extracts suggested an increase in structural glycoprotein content and a decrease in collagen solubility in the long bones of mutants. Significant differences were found in the organic matrix composition of male and female bones of the two strains. /sup 14/C-glucose incorporation was stronger in males than in females. The presence of a glycosaminoglycan different from the chondroitinesulfate was shown in males. Basic amino acid content (lysine, arginine, histidine) was clearly higher in the insoluble residue of male bones .

Biochemical studies of the macromolecular matrix of long bones in the Op/Orl mutant rat strain

International Nuclear Information System (INIS)

Moczar, E.; Berenholc, S.; Phan-Dinh-Tuy, B.; Robert, A.M.

1978-01-01

The long bones of normal and Op/Orl mutant rats were incubated with 14 C-glucose and fractionated by EDTA and urea extraction. The analytical results of the various extracts suggested an increase in structural glycoprotein content and a decrease in collagen solubility in the long bones of mutants. Significant differences were found in the organic matrix composition of male and female bones of the two strains. 14 C-glucose incorporation was stronger in males than in females. The presence of a glycosaminoglycan different from the chondroitinesulfate was shown in males. Basic amino acid content (lysine, arginine, histidine) was clearly higher in the insoluble residue of male bones

Raman and Fourier Transform Infrared (FT-IR) Mineral to Matrix Ratios Correlate with Physical Chemical Properties of Model Compounds and Native Bone Tissue.

Science.gov (United States)

Taylor, Erik A; Lloyd, Ashley A; Salazar-Lara, Carolina; Donnelly, Eve

2017-10-01

Raman and Fourier transform infrared (FT-IR) spectroscopic imaging techniques can be used to characterize bone composition. In this study, our objective was to validate the Raman mineral:matrix ratios (ν 1 PO 4 :amide III, ν 1 PO 4 :amide I, ν 1 PO 4 :Proline + hydroxyproline, ν 1 PO 4 :Phenylalanine, ν 1 PO 4 :δ CH 2 peak area ratios) by correlating them to ash fraction and the IR mineral:matrix ratio (ν 3 PO 4 :amide I peak area ratio) in chemical standards and native bone tissue. Chemical standards consisting of varying ratios of synthetic hydroxyapatite (HA) and collagen, as well as bone tissue from humans, sheep, and mice, were characterized with confocal Raman spectroscopy and FT-IR spectroscopy and gravimetric analysis. Raman and IR mineral:matrix ratio values from chemical standards increased reciprocally with ash fraction (Raman ν 1 PO 4 /Amide III: P Raman ν 1 PO 4 /Amide I: P Raman ν 1 PO 4 /Proline + Hydroxyproline: P Raman ν 1 PO 4 /Phenylalanine: P Raman ν 1 PO 4 /δ CH 2 : P Raman and IR mineral:matrix ratio values were strongly correlated ( P Raman mineral:matrix bone composition parameter correlates strongly to ash fraction and to its IR counterpart. Finally, the mineral:matrix ratio values of the native bone tissue are similar to those of both chemical standards and theoretical values, confirming the biological relevance of the chemical standards and the characterization techniques.

Biosynthesis and Characterization of Nanocellulose-Gelatin Films

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Muenduen Phisalaphong

2013-02-01

Full Text Available A nanocellulose-gelatin (bacterial cellulose gelatin (BCG film was developed by a supplement of gelatin, at a concentration of 1%–10% w/v, in a coconut-water medium under the static cultivation of Acetobacter xylinum. The two polymers exhibited a certain degree of miscibility. The BCG film displayed dense and uniform homogeneous structures. The Fourier transform infrared spectroscopy (FTIR results demonstrated interactions between the cellulose and gelatin. Incorporation of gelatin into a cellulose nanofiber network resulted in significantly improved optical transparency and water absorption capacity of the films. A significant drop in the mechanical strengths and a decrease in the porosity of the film were observed when the supplement of gelatin was more than 3% (w/v. The BCG films showed no cytotoxicity against Vero cells.

Instrumental texture profile analysis of gelatin gel extracted from grouper skin and commercial (bovine and porcine) gelatin gels.

Science.gov (United States)

Rahman, Mohammad Shafiur; Al-Mahrouqi, Abdullah Issa

2009-01-01

Mechanical compression was used to study the gelling characteristics of gelatin gels. Texture profile analysis (TPA) showed that the hardness of fish and mammalian gelatin increased significantly as the concentrations of gels increased. TPA attributes of 10% fish skin gel showed significant differences from those obtained from 20% and 30% gels. In bovine and porcine cases, such generic trends were not observed. Mechanical characteristics of 10% gels of gelatin from fish skin, determined from one cycle compression, were significantly lower than other sources of gelatin gels, while bovine and porcine gels did not show any significant differences. In the case of TPA, hardness of bovine gelatin gel was highest at 41 N for 10% gel, followed by porcine (30 N) then fish skin (5 N) gelatin gels. The gels prepared from different sources did not show any generic trends when all other mechanical attributes were considered.

Construction of human induced pluripotent stem cell-derived oriented bone matrix microstructure by using in vitro engineered anisotropic culture model.

Science.gov (United States)

Ozasa, Ryosuke; Matsugaki, Aira; Isobe, Yoshihiro; Saku, Taro; Yun, Hui-Suk; Nakano, Takayoshi

2018-02-01

Bone tissue has anisotropic microstructure based on collagen/biological apatite orientation, which plays essential roles in the mechanical and biological functions of bone. However, obtaining an appropriate anisotropic microstructure during the bone regeneration process remains a great challenging. A powerful strategy for the control of both differentiation and structural development of newly-formed bone is required in bone tissue engineering, in order to realize functional bone tissue regeneration. In this study, we developed a novel anisotropic culture model by combining human induced pluripotent stem cells (hiPSCs) and artificially-controlled oriented collagen scaffold. The oriented collagen scaffold allowed hiPSCs-derived osteoblast alignment and further construction of anisotropic bone matrix which mimics the bone tissue microstructure. To the best of our knowledge, this is the first report showing the construction of bone mimetic anisotropic bone matrix microstructure from hiPSCs. Moreover, we demonstrated for the first time that the hiPSCs-derived osteoblasts possess a high level of intact functionality to regulate cell alignment. © 2017 The Authors Journal of Biomedical Materials Research Part A Published by Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 360-369, 2018. © 2017 The Authors Journal of Biomedical Materials Research Part A Published by Wiley Periodicals, Inc.

Silver-gelatine bionanocomposites for qualitative detection of a pesticide by SERS.

Science.gov (United States)

Fateixa, S; Soares, S F; Daniel-da-Silva, A L; Nogueira, H I S; Trindade, T

2015-03-07

The controlled release of pesticides using hydrogel vehicles is an important procedure to limit the amount of these compounds in the environment, providing an effective way for crop protection. A key-step in the formulation of new materials for these purposes encompasses the monitoring of available pesticides in the gel matrix under variable working conditions. In this work, we report a series of bionanocomposites made of Ag nanoparticles (NPs) and gelatine A for the surface enhanced Raman scattering (SERS) detection of sodium diethyldithiocarbamate (EtDTC) as a pesticide model. These studies demonstrate the effectiveness of these substrates for the detection of EtDTC in aqueous solutions in a concentration as low as 10(-5) M. We have monitored the Raman signal enhancement of this analyte in bionanocomposites having an increasing amount of gelatine due to their relevance in formulating hydrogels of variable gel strengths. Under these conditions, the bionanocomposites have shown an effective SERS activity using EtDTC, demonstrating their effectiveness in the qualitative detection of this analyte. Finally, experiments involving the release of EtDTC from Ag/gelatine samples have been monitored by SERS, which attest the potential of this spectroscopic method in the laboratorial monitoring of hydrogels for pesticide release.

Enzymatic degradation of polycaprolactone–gelatin blend

International Nuclear Information System (INIS)

Banerjee, Aditi; Chatterjee, Kaushik; Madras, Giridhar

2015-01-01

Blends of polycaprolactone (PCL), a synthetic polymer and gelatin, natural polymer offer a optimal combination of strength, water wettability and cytocompatibility for use as a resorbable biomaterial. The enzymatic degradation of PCL, gelatin and PCL–gelatin blended films was studied in the presence of lipase (Novozym 435, immobilized) and lysozyme. Novozym 435 degraded the PCL films whereas lysozyme degraded the gelatin. Though Novozym 435 and lysozyme individually could degrade PCL–gelatin blended films, the combination of these enzymes showed the highest degradation of these blended films. Moreover, the enzymatic degradation was much faster when fresh enzymes were added at regular intervals. The changes in physico-chemical properties of polymer films due to degradation were studied by scanning electron microscopy, Fourier transform infrared spectroscopy and differential scanning calorimetry. These results have important implications for designing resorbable biomedical implants. (paper)

Interaction of gelatin with polyenes modulates antifungal activity and biocompatibility of electrospun fiber mats

Directory of Open Access Journals (Sweden)

Lakshminarayanan R

2014-05-01

Full Text Available Rajamani Lakshminarayanan,1,2 Radhakrishnan Sridhar,3,4 Xian Jun Loh,5 Muruganantham Nandhakumar,1 Veluchamy Amutha Barathi,1,6 Madhaiyan Kalaipriya,3,4 Jia Lin Kwan,1 Shou Ping Liu,1,2 Roger Wilmer Beuerman,1,2 Seeram Ramakrishna3,4,7 1Singapore Eye Research Institute, 2Signature Research Program in Neuroscience and Behavioral Disorders, Duke-NUS Graduate Medical School, 3Department of Mechanical Engineering, National University of Singapore, 4Center for Nanofibers and Nanotechnology, National University of Singapore, 5Institute of Materials Research and Engineering, A*STAR (Agency for Science, Technology and Research, 3 Research Link, Singapore, 6Department of Ophthalmology, Yong Loo Lin School of Medicine, National University of Singapore, 7NUS Nanoscience and Nanotechnology Initiative, Singapore Abstract: Topical application of antifungals does not have predictable or well-controlled release characteristics and requires reapplication to achieve therapeutic local concentration in a reasonable time period. In this article, the efficacy of five different US Food and Drug Administration-approved antifungal-loaded (amphotericin B, natamycin, terbinafine, fluconazole, and itraconazole electrospun gelatin fiber mats were compared. Morphological studies show that incorporation of polyenes resulted in a two-fold increase in fiber diameter and the mats inhibit the growth of yeasts and filamentous fungal pathogens. Terbinafine-loaded mats were effective against three filamentous fungal species. Among the two azole antifungals compared, the itraconazole-loaded mat was potent against Aspergillus strains. However, activity loss was observed for fluconazole-loaded mats against all of the test organisms. The polyene-loaded mats displayed rapid candidacidal activities as well. Biophysical and rheological measurements indicate strong interactions between polyene antifungals and gelatin matrix. As a result, the polyenes stabilized the triple helical

Anaphylaxis to gelatin-containing rectal suppositories.

Science.gov (United States)

Sakaguchi, M; Inouye, S

2001-12-01

Some children--though the number is few-have been sensitized with gelatin. To investigate the relationship between the presence of antigelatin IgE and anaphylaxis to gelatin-containing rectal suppository, we measured antigelatin IgE in the sera of the children with anaphylaxis. Ten children showed systemic allergic reactions, including anaphylaxis, to a chloral hydrate rectal suppository containing gelatin (231 mg/dose) that had been used as a sedative. These children's clinical histories and serum samples were submitted from physicians to the National Institute of Infectious Diseases during a 2-year period from 1996 to 1997. Of the 10 children, 5 showed apparent anaphylaxis, including hypotension and/or cyanosis, along with urticaria or wheezing; 2 showed both urticaria and wheezing without hypotension or cyanosis; the other 3 showed only urticaria. All of the children had antigelatin IgE (mean value +/- SD, 7.9 +/- 8.4 Ua/mL). As a control, samples from 250 randomly selected children had no antigelatin IgE. These findings suggest that the 10 children's systemic allergic reactions to this suppository were caused by the gelatin component. Gelatin-containing suppositories must be used with the same caution as gelatin-containing vaccines and other medications.

Bi-layered nanocomposite bandages for controlling microbial infections and overproduction of matrix metalloproteinase activity.

Science.gov (United States)

Anjana, J; Mohandas, Annapoorna; Seethalakshmy, S; Suresh, Maneesha K; Menon, Riju; Biswas, Raja; Jayakumar, R

2018-04-15

Chronic diabetic wounds is characterised by increased microbial contamination and overproduction of matrix metalloproteases that would degrade the extracellular matrix. A bi-layer bandage was developed, that promotes the inhibition of microbial infections and matrix metalloprotease (MMPs) activity. Bi-layer bandage containing benzalkonium chloride loaded gelatin nanoparticles (BZK GNPs) in chitosan-Hyaluronic acid (HA) as a bottom layer and sodium alendronate containing chitosan as top layer was developed. We hypothesized that the chitosan-gelatin top layer with sodium alendronate could inhibit the MMPs activity, whereas the chitosan-HA bottom layer with BZK GNPs (240±66nm) would enable the elimination of microbes. The porosity, swelling and degradation nature of the prepared Bi-layered bandage was studied. The bottom layer could degrade within 4days whereas the top layer remained upto 7days. The antimicrobial activity of the BZK NPs loaded bandage was determined using normal and clinical strains. Gelatin zymography shows that the proteolytic activity of MMP was inhibited by the bandage. Copyright © 2017 Elsevier B.V. All rights reserved.

Effects of increased collagen-matrix density on the mechanical properties and in vivo absorbability of hydroxyapatite-collagen composites as artificial bone materials

Energy Technology Data Exchange (ETDEWEB)

Yunoki, Shunji [Life Science Group, Tokyo Metropolitan Industrial Technology Research Institute, 2-11-1 Fukasawa, Setagaya-ku, Tokyo 158-0081 (Japan); Sugiura, Hiroaki; Kondo, Eiji; Yasuda, Kazunori [Department of Sports Medicine and Joint Surgery, Graduate School of Medicine, Hokkaido University, Kita-15 Nishi-7, Sapporo, Hokkaido 060-8638 Japan (Japan); Ikoma, Toshiyuki; Tanaka, Junzo, E-mail: yunoki.shunji@iri-tokyo.jp [Department of Metallurgy and Ceramics Science, 2-12-1-S7-1, Ookayama, Meguro-ku, Tokyo 152-8550 (Japan)

2011-02-15

The aim of this study was to evaluate the effects of increased collagen-matrix density on the mechanical properties and in vivo absorbability of porous hydroxyapatite (HAp)-collagen composites as artificial bone materials. Seven types of porous HAp-collagen composites were prepared from HAp nanocrystals and dense collagen fibrils. Their densities and HAp/collagen weight ratios ranged from 122 to 331 mg cm{sup -3} and from 20/80 to 80/20, respectively. The flexural modulus and strength increased with an increase in density, reaching 2.46 {+-} 0.48 and 0.651 {+-} 0.103 MPa, respectively. The porous composites with a higher collagen-matrix density exhibited much higher mechanical properties at the same densities, suggesting that increasing the collagen-matrix density is an effective way of improving the mechanical properties. It was also suggested that other structural factors in addition to collagen-matrix density are required to achieve bone-like mechanical properties. The in vivo absorbability of the composites was investigated in bone defects of rabbit femurs, demonstrating that the absorption rate decreased with increases in the composite density. An exhaustive increase in density is probably limited by decreases in absorbability as artificial bones.

Effects of increased collagen-matrix density on the mechanical properties and in vivo absorbability of hydroxyapatite-collagen composites as artificial bone materials

International Nuclear Information System (INIS)

Yunoki, Shunji; Sugiura, Hiroaki; Kondo, Eiji; Yasuda, Kazunori; Ikoma, Toshiyuki; Tanaka, Junzo

2011-01-01

The aim of this study was to evaluate the effects of increased collagen-matrix density on the mechanical properties and in vivo absorbability of porous hydroxyapatite (HAp)-collagen composites as artificial bone materials. Seven types of porous HAp-collagen composites were prepared from HAp nanocrystals and dense collagen fibrils. Their densities and HAp/collagen weight ratios ranged from 122 to 331 mg cm -3 and from 20/80 to 80/20, respectively. The flexural modulus and strength increased with an increase in density, reaching 2.46 ± 0.48 and 0.651 ± 0.103 MPa, respectively. The porous composites with a higher collagen-matrix density exhibited much higher mechanical properties at the same densities, suggesting that increasing the collagen-matrix density is an effective way of improving the mechanical properties. It was also suggested that other structural factors in addition to collagen-matrix density are required to achieve bone-like mechanical properties. The in vivo absorbability of the composites was investigated in bone defects of rabbit femurs, demonstrating that the absorption rate decreased with increases in the composite density. An exhaustive increase in density is probably limited by decreases in absorbability as artificial bones.

Neutrophil elastase processing of Gelatinase A is mediated by extracellular matrix

Energy Technology Data Exchange (ETDEWEB)

Rice, A.; Banda, M.J. [Univ. of California, San Franciso, CA (United States)

1995-07-18

Gelatinase A (72-kDa type IV collagenase) is a metalloproteinase that is expressed by many cells in culture and is overexpressed by some tumor cells. It has been suggested that the serine proteinase neutrophil elastase might play a role iii the posttranslational processing of gelatinase A and that noncatalytic interactions between gelatinase A and components of the extracellular matrix might alter potential processing pathways. These questions were addressed with the use of gelatin substrate zymography, gelatinolytic activity assays, and amino acid sequence analysis. We found that neutrophil elastase does proteolytically modify gelatinase A by cleaving at a number of sites within gelatinase A. Sequential treatment of gelatinase A with 4-aminophenylmercuric acetate (APMA) and neutrophil elastase yielded an active gelatinase with a 4-fold increase in gelatinolytic activity. The increased gelatinolytic activity correlated with that of a 40-kDa fragment of gelatinase A. Matrix components altered the proteolytic modifications in gelatinase A that were mediated by neutrophil elastase. In the absence of gelatin, neutrophil elastase destructively degraded gelatinase A by hydrolyzing at least two bonds within the fibronectin-like gelatin-binding domain of gelatinase A. In the presence of gelatin, these two inactivating cleavage sites were protected, and cleavage at a site within the hemopexin-like carboxyl-terminal domain resulted in a truncated yet active gelatinase. The results suggest a regulatory role for extracellular matrix molecules in stabilizing gelatinase A fragments and in altering the availability of sites susceptible to destructive proteolysis by neutrophil elastase. 32 refs., 10 figs.

Closing the medullary canal after retrograde nail removal using a bioabsorbable bone plug: technical tip

NARCIS (Netherlands)

Schepers, T.; Vogels, L. M. M.

2012-01-01

We describe a simple technique for closure of the intra-articular opening after the removal of a retrograde femur nail. With the use of a gelatine bioabsorbable bone plug the medullary canal is closed, reducing leakage of blood and cancellous bone particles from the bone into the knee joint

Molecular interactions in gelatin/chitosan composite films.

Science.gov (United States)

Qiao, Congde; Ma, Xianguang; Zhang, Jianlong; Yao, Jinshui

2017-11-15

Gelatin and chitosan were mixed at different mass ratios in solution forms, and the rheological properties of these film-forming solutions, upon cooling, were studied. The results indicate that the significant interactions between gelatin and chitosan promote the formation of multiple complexes, reflected by an increase in the storage modulus of gelatin solution. Furthermore, these molecular interactions hinder the formation of gelatin networks, consequently decreasing the storage modulus of polymer gels. Both hydrogen bonds and electrostatic interactions are formed between gelatin and chitosan, as evidenced by the shift of the amide-II bands of polymers. X-ray patterns of composite films indicate that the contents of triple helices decrease with increasing chitosan content. Only one glass transition temperature (T g ) was observed in composite films with different composition ratios, and it decreases gradually with an increase in chitosan proportion, indicating that gelatin and chitosan have good miscibility and form a wide range of blends. Copyright © 2017 Elsevier Ltd. All rights reserved.

Preservation and storage of prepared ballistic gelatine.

Science.gov (United States)

Mattijssen, E J A T; Alberink, I; Jacobs, B; van den Boogaard, Y

2016-02-01

The use of ballistic gelatine, generally accepted as a human muscle tissue simulant in wound ballistic studies, might be improved by adding a preservative (Methyl 4-hydroxybenzoate) which inhibits microbial growth. This study shows that replacing a part of the gelatine powder by the preservative does not significantly alter the penetration depth of projectiles. Storing prepared blocks of ballistic gelatine over time decreased the penetration depth of projectiles. Storage of prepared gelatine for 4 week already showed a significant effect on the penetration depth of projectiles. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Influence of Porcine Intervertebral Disc Matrix on Stem Cell Differentiation

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Hans-Lothar Fuchsbauer

2011-08-01

Full Text Available For back disorders, cell therapy is one approach for a real regeneration of a degenerated nucleus pulposus. Human mesenchymal stem cells (hMSC could be differentiated into nucleus pulposus (NP-like cells and used for cell therapy. Therefore it is necessary to find a suitable biocompatible matrix, which supports differentiation. It could be shown that a differentiation of hMSC in a microbial transglutaminase cross-linked gelatin matrix is possible, but resulted in a more chondrocyte-like cell type. The addition of porcine NP extract to the gelatin matrix caused a differentiation closer to the desired NP cell phenotype. This concludes that a hydrogel containing NP extract without any other supplements could be suitable for differentiation of hMSCs into NP cells. The NP extract itself can be cross-linked by transglutaminase to build a hydrogel free of NP atypical substrates. As shown by side-specific biotinylation, the NP extract contains molecules with free glutamine and lysine residues available for the transglutaminase.

Characterization of Fish Skin Gelatin Yellowfin Tuna

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Mala Nurilmala

2017-08-01

Full Text Available Gelatin is one of the products which become a necessity for various industries, i.e. food and non-food industries. The application of gelatin has been increasing year by year in Indonesia. However, there is nogelatin industry in Indonesia so far. Thus, it is necessary to find an alternative source of gelatin, especially from fishery by products.Thus, the purpose of this research was to extract fish skin gelatin of yellowfin tuna with temperature treatments (55, 65 and 75oC. In addition, the properties of resulted gelatin were determined including yield, pH, gel strength, viscosity, functional groups, molecular weight profiles, and amino acid composition. The extraction at 75oC was chosen as the best result. The yield was 17%; pH 5.3; gel strength 1789.55 gf, viscosity 104.2 Cp, respectively. There was functional group amide A, I, II, dan III. SDS-PAGE showed β, α1 dan α2 bands for tuna skin gelatin. In addition, the main amino acids were glycine and proline.

Preparation and comparative characterization of keratin–chitosan and keratin–gelatin composite scaffolds for tissue engineering applications

International Nuclear Information System (INIS)

Balaji, S.; Kumar, Ramadhar; Sripriya, R.; Kakkar, Prachi; Ramesh, D. Vijaya; Reddy, P. Neela Kanta; Sehgal, P.K.

2012-01-01

We report fabrication of three dimensional scaffolds with well interconnected matrix of high porosity using keratin, chitosan and gelatin for tissue engineering and other biomedical applications. Scaffolds were fabricated using porous Keratin–Gelatin (KG), Keratin–Chitosan (KC) composites. The morphology of both KG and KC was investigated using SEM. The scaffolds showed high porosity with interconnected pores in the range of 20–100 μm. They were further tested by FTIR, DSC, CD, tensile strength measurement, water uptake and swelling behavior. In vitro cell adhesion and cell proliferation tests were carried out to study the biocompatibility behavior and their application as an artificial skin substitute. Both KG and KC composite scaffolds showed similar properties and patterns for cell proliferation. Due to rapid degradation of gelatin in KG, we found that it has limited application as compared to KC scaffold. We conclude that KC scaffold owing to its slow degradation and antibacterial properties would be a better substrate for tissue engineering and other biomedical application. Highlights: ► Extraction of reduced keratin from horn meal. ► Preparation of keratin–gelatin and keratin–chitosan composite scaffolds. ► Characterizations of the composite scaffolds. ► Comparative cytotoxicity analysis on NIH3T3 fibroblasts.

Polymer gel dosimeter with AQUAJOINT® as hydrogel matrix

Science.gov (United States)

Maeyama, Takuya; Ishida, Yasuhiro; Kudo, Yoshihiro; Fukasaku, Kazuaki; Ishikawa, Kenichi L.; Fukunishi, Nobuhisa

2018-05-01

We report a polymer gel dosimeter based on a new gel matrix (AQUAJOINT®) that is a thermo-irreversible hydrogel formed by mixing two types of water-based liquids at room temperature. Normoxic N-vinylpyrrolidone-based polymer gels were prepared with AQUAJOINT® instead of gelatin. This AQUAJOINT®-based gel dosimeter exhibits a 2.5-fold increase in sensitivity over a gelatin-based gel dosimeter and a linear dose-response in the dose range of 0-8 Gy. This gel has heat resistance in a jar and controlled gel properties such as viscoelastic and mechanical characters, which may be useful for deformable polymer gel dosimetry.

The available evidence on demineralised bone matrix in trauma and orthopaedic surgery : a systematic review

NARCIS (Netherlands)

van der Stok, J.; Hartholt, K.A.; Schoenmakers, D.A.L.; Arts, J.J.C.

2017-01-01

Objectives: The aim of this systematic literature review was to assess the clinical level of evidence of commercially available demineralised bone matrix (DBM) products for their use in trauma and orthopaedic related surgery. Methods: A total of 17 DBM products were used as search terms in two

Direct Scaffolding of Biomimetic Hydroxyapatite-gelatin Nanocomposites using Aminosilane Cross-linker for Bone Regeneration

OpenAIRE

Chiu, Chi-Kai; Ferreira, Joao; Luo, Tzy-Jiun M.; Geng, Haixia; Lin, Feng-Chang; Ko, Ching-Chang

2012-01-01

Hydroxyapatite-gelatin modified siloxane (GEMOSIL) nanocomposite was developed by coating, kneading and hardening processes to provide formable scaffolding for alloplastic graft applications. The present study aims to characterize scaffolding formability and mechanical properties of GEMOSIL, and to test the in vitro and in vivo biocompatibility of GEMOSIL. Buffer Solution initiated formable paste followed by the sol-gel reaction led to a final hardened composite. Results showed the adequate c...

The fabrication of nanocomposites via calcium phosphate formation on gelatin–chitosan network and the gelatin influence on the properties of biphasic composites

International Nuclear Information System (INIS)

Babaei, Zahra; Jahanshahi, Mohsen; Rabiee, Sayed Mahmood

2013-01-01

A novel biodegradable polymer–ceramic nanocomposite which consisted of gelatin (Gel), chitosan (CS), and calcium phosphate (CaP) nanoparticles was prepared based on in situ preparation method. The fabricated biocomposites were characterized by FTIR, X-ray diffraction (XRD), transmission electron microscopy (TEM) as well as scanning electron microscope with X-ray elemental analysis (SEM-EDX). The characterization results confirmed that the crystalline calcium phosphate nanoparticles were mineralized in polymeric matrix and the interaction between Ca2+ in calcium phosphate and functional groups in polymers molecular chains was formed. XRD result showed that in addition to hydroxyapatite (HA), Brushite (BR) and tricalcium phosphate (β-TCP) particles also were formed due to lack of complete penetration of the basic solution into the polymeric matrix. However, SEM image indicated that the polymeric matrix has the controlling role in the particle size of calcium phosphate. The size of particles in three component composites was about 100 nm while in two component composites proved to be more in μm size. TEM observation supported SEM results and showed that the three component composites have calcium phosphate nanoparticles. The elastic modulus and compressive strength of the composites were also improved by the employment of gelatin and chitosan together, which can make them more beneficial for surgical applications. - Highlights: ► A new type of calcium phosphate nanocomposites consisted chitosan and gelatin as polymeric matrix was obtained via in situ precipitation method. ► The formed particles into the polymeric matrix are hydroxyapatite, Brushite and β-tricalcium phosphate. ► The polymer concentration is a predominant factor for inhibiting nucleation and growth of calcium phosphate particles as the particles size was also decreased by increasing the amount of polymeric phase. ► The nano particles formed in three component composites and micro particles

The role of surface charge in the desolvation process of gelatin: implications in nanoparticle synthesis and modulation of drug release

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Ahsan SM

2017-01-01

Full Text Available Saad M Ahsan, Chintalagiri Mohan Rao Centre for Cellular and Molecular Biology, Council of Scientific and Industrial Research, Hyderabad, Telangana, India Abstract: The process of moving hydrophobic amino acids into the core of a protein by desolvation is important in protein folding. However, a rapid and forced desolvation can lead to precipitation of proteins. Desolvation of proteins under controlled conditions generates nanoparticles – homogeneous aggregates with a narrow size distribution. The protein nanoparticles, under physiological conditions, undergo surface erosion due to the action of proteases, releasing the entrapped drug/gene. The packing density of protein nanoparticles significantly influences the release kinetics. We have investigated the desolvation process of gelatin, exploring the role of pH and desolvating agent in nanoparticle synthesis. Our results show that the desolvation process, initiated by the addition of acetone, follows distinct pathways for gelatin incubated at different pH values and results in the generation of nanoparticles with varying matrix densities. The nanoparticles synthesized with varying matrix densities show variations in drug loading and protease-dependent extra- and intracellular drug release. These results will be useful in fine-tuning the synthesis of nanoparticles with desirable drug release profiles. Keywords: protein desolvation, nanoparticle assembly, gelatin nanoparticle synthesis, protease susceptibility, intracellular drug release

Extraction and electrospinning of gelatin from fish skin.

Science.gov (United States)

Songchotikunpan, Panida; Tattiyakul, Jirarat; Supaphol, Pitt

2008-04-01

Ultra-fine gelatin fibers were successfully fabricated by electrospinning from the solutions of Nile tilapia (Oreochromis niloticus) skin-extracted gelatin in either acetic acid or formic acid aqueous solutions. The extracted gelatin contained 7.3% moisture, 89.4% protein, 0.3% lipid, and 0.4% ash contents (on the basis of wet weight), while the bloom gel strength, the shear viscosity, and the pH values were 328 g, 17.8 mPa s, and 5.0, respectively. Both the acid concentration and the concentration of the gelatin solutions strongly influenced the properties of the as-prepared solutions and the obtained gelatin fibers. At low acid concentrations (i.e., 15% (w/v) extracted gelatin solutions in 10 and 20% (v/v) acetic acid solvents or 10-60% (v/v) formic acid solvents), a combination between smooth and beaded fibers was observed. At low concentrations of the gelatin solutions in either 40% (v/v) acetic acid solvent or 80% (v/v) formic acid solvent (i.e., 5-11%, w/v), either discrete beads or beaded fibers were obtained, while, at higher concentrations (i.e., 14-29%, w/v), only smooth or a combination of smooth and beaded fibers were obtained. The average diameters of the obtained fibers, regardless of the types of the acid solvents used, ranged between 109 and 761 nm. Lastly, cross-linking of the obtained gelatin fiber mats with glutaraldehyde vapor caused slight shrinkage from their original dimension, and the cross-linked gelatin fiber mats became stiffer.

3D Bioprinting of Heterogeneous Aortic Valve Conduits with Alginate/Gelatin Hydrogels

Science.gov (United States)

Duan, Bin; Hockaday, Laura A.; Kang, Kevin H.; Butcher, Jonathan T.

2013-01-01

Heart valve disease is a serious and growing public health problem for which prosthetic replacement is most commonly indicated. Current prosthetic devices are inadequate for younger adults and growing children. Tissue engineered living aortic valve conduits have potential for remodeling, regeneration, and growth, but fabricating natural anatomical complexity with cellular heterogeneity remain challenging. In the current study, we implement 3D bioprinting to fabricate living alginate/gelatin hydrogel valve conduits with anatomical architecture and direct incorporation of dual cell types in a regionally constrained manner. Encapsulated aortic root sinus smooth muscle cells (SMC) and aortic valve leaflet interstitial cells (VIC) were viable within alginate/gelatin hydrogel discs over 7 days in culture. Acellular 3D printed hydrogels exhibited reduced modulus, ultimate strength, and peak strain reducing slightly over 7-day culture, while the tensile biomechanics of cell-laden hydrogels were maintained. Aortic valve conduits were successfully bioprinted with direct encapsulation of SMC in the valve root and VIC in the leaflets. Both cell types were viable (81.4±3.4% for SMC and 83.2±4.0% for VIC) within 3D printed tissues. Encapsulated SMC expressed elevated alpha-smooth muscle actin when printed in stiff matrix, while VIC expressed elevated vimentin in soft matrix. These results demonstrate that anatomically complex, heterogeneously encapsulated aortic valve hydrogel conduits can be fabricated with 3D bioprinting. PMID:23015540

In vitro and in vivo evaluation of chitosan–gelatin scaffolds for cartilage tissue engineering

Energy Technology Data Exchange (ETDEWEB)

Whu, Shu Wen [Department of Orthopaedic Surgery, Chang Gung Memorial Hospital at Keelung, College of Medicine, Chang Gung University, Taoyuan, Taiwan (China); Hung, Kun-Che; Hsieh, Kuo-Huang [Institute of Polymer Science and Engineering, National Taiwan University, Taipei, Taiwan (China); Chen, Chih-Hwa [Department of Orthopaedic Surgery, Chang Gung Memorial Hospital at Keelung, College of Medicine, Chang Gung University, Taoyuan, Taiwan (China); Tsai, Ching-Lin, E-mail: tsaicl@ntuh.gov.tw [Department of Orthopaedics, National Taiwan University Hospital, Taipei, Taiwan (China); Hsu, Shan-hui, E-mail: shhsu@ntu.edu.tw [Institute of Polymer Science and Engineering, National Taiwan University, Taipei, Taiwan (China)

2013-07-01

Chitosan–gelatin polyelectrolyte complexes were fabricated and evaluated as tissue engineering scaffolds for cartilage regeneration in vitro and in vivo. The crosslinker for the gelatin component was selected among glutaraldehyde, bisepoxy, and a water-soluble carbodiimide (WSC) based upon the proliferation of chondrocytes on the crosslinked gelatin. WSC was found to be the most suitable crosslinker. Complex scaffolds made from chitosan and gelatin with a component ratio equal to one possessed the proper degradation rate and mechanical stability in vitro. Chondrocytes were able to proliferate well and secrete abundant extracellular matrix in the chitosan–gelatin (1:1) complex scaffolds crosslinked by WSC (C1G1{sub WSC}) compared to the non-crosslinked scaffolds. Implantation of chondrocytes-seeded scaffolds in the defects of rabbit articular cartilage confirmed that C1G1{sub WSC} promoted the cartilage regeneration. The neotissue formed the histological feature of tide line and lacunae in 6.5 months. The amount of glycosaminoglycans in C1G1{sub WSC} constructs (0.187 ± 0.095 μg/mg tissue) harvested from the animals after 6.5 months was 14 wt.% of that in normal cartilage (1.329 ± 0.660 μg/mg tissue). The average compressive modulus of regenerated tissue at 6.5 months was about 0.539 MPa, which approached to that of normal cartilage (0.735 MPa), while that in the blank control (3.881 MPa) was much higher and typical for fibrous tissue. Type II collagen expression in C1G1{sub WSC} constructs was similarly intense as that in the normal hyaline cartilage. According to the above results, the use of C1G1{sub WSC} scaffolds may enhance the cartilage regeneration in vitro and in vivo. - Highlights: • We developed a chitosan–gelatin scaffold crosslinked with carbodiimide. • Neocartilage formation was more evident in crosslinked vs. non-crosslinked scaffolds. • Histological features of tide line and lacunae were observed in vivo at 6.5 months. • Compressive

In vivo performance of combinations of autograft, demineralized bone matrix, and tricalcium phosphate in a rabbit femoral defect model

International Nuclear Information System (INIS)

Kim, Jinku; McBride, Sean; Hollinger, Jeffrey O; Dean, David D; Sylvia, Victor L; Doll, Bruce A

2014-01-01

Large bone defects may be treated with autologous or allogeneic bone preparations. Each treatment has advantages and disadvantages; therefore, a clinically viable option for treating large (e.g., gap) bone defects may be a combination of the two. In the present study, bone repair was determined with combinations of autografts, allografts, and synthetic bone grafts using an established rabbit femoral defect model. Bilateral unicortical femoral defects were surgically prepared and treated with combinatorial bone grafts according to one of seven treatment groups. Recipient sites were retrieved at six weeks. Cellular/tissue responses and new bone formation were assessed by histology and histomorphometry. Histological analysis images indicated neither evidence of inflammatory, immune responses, tissue necrosis, nor osteolysis. Data suggested co-integration of implanted agents with host and newly formed bone. Finally, the histomorphometric data suggested that the tricalcium phosphate-based synthetic bone graft substitute allowed new bone formation that was similar to the allograft (i.e., demineralized bone matrix, DBM). (paper)

Time domain optical coherence tomography investigation of bone matrix interface in rat femurs

Science.gov (United States)

Rusu, Laura-Cristina; Negruá¹±iu, Meda-Lavinia; Sinescu, Cosmin; Hoinoiu, Bogdan; Topala, Florin-Ionel; Duma, Virgil-Florin; Rominu, Mihai; Podoleanu, Adrian G.

2013-08-01

The materials used to fabricate scaffolds for tissue engineering are derived from synthetic polymers, mainly from the polyester family, or from natural materials (e.g., collagen and chitosan). The mechanical properties and the structural properties of these materials can be tailored by adjusting the molecular weight, the crystalline state, and the ratio of monomers in the copolymers. Quality control and adjustment of the scaffold manufacturing process are essential to achieve high standard scaffolds. Most scaffolds are made from highly crystalline polymers, which inevitably result in their opaque appearance. Their 3-D opaque structure prevents the observation of internal uneven surface structures of the scaffolds under normal optical instruments, such as the traditional light microscope. The inability to easily monitor the inner structure of scaffolds as well as the interface with the old bone poses a major challenge for tissue engineering: it impedes the precise control and adjustment of the parameters that affect the cell growth in response to various mimicked culture conditions. The aim of this paper is to investigate the interface between the femur rat bone and the new bone that is obtained using a method of tissue engineering that is based on different artificial matrixes inserted in previously artificially induced defects. For this study, 15 rats were used in conformity with ethical procedures. In all the femurs a round defect was induced by drilling with a 1 mm spherical Co-Cr surgical drill. The matrixes used were Bioss and 4bone. These materials were inserted into the induced defects. The femurs were investigated at 1 week, 1 month, 2 month and three month after the surgical procedures. The interfaces were examined using Time Domain (TD) Optical Coherence Tomography (OCT) combined with Confocal Microscopy (CM). The optical configuration uses two single mode directional couplers with a superluminiscent diode as the source centered at 1300 nm. The scanning

Role of solvent-mediated carbodiimide cross-linking in fabrication of electrospun gelatin nanofibrous membranes as ophthalmic biomaterials

International Nuclear Information System (INIS)

Chou, Shih-Feng; Luo, Li-Jyuan; Lai, Jui-Yang; Ma, David Hui-Kang

2017-01-01

Due to their ability to mimic the structure of extracellular matrix, electrospun gelatin nanofibers are promising cell scaffolding materials for tissue engineering applications. However, the hydrophilic gelatin molecules usually need stabilization before use in aqueous physiological environment. Considering that biomaterials cross-linked via film immersion technique may have a more homogeneous cross-linked structure than vapor phase cross-linking, this work aims to investigate the chemical modification of electrospun gelatin nanofibrous membranes by liquid phase carbodiimide in the presence of ethanol/water co-solvents with varying ethanol concentrations ranging from 80 to 99.5 vol%. The results of characterization showed that increasing water content in the binary reaction solvent system increases the extent of cross-linking of gelatin nanofibers, but simultaneously promotes the effect of biopolymer swelling and distortion in fiber mat structure. As compared to non-cross-linked counterparts, carbodiimide treated gelatin nanofibrous mats exhibited better thermal and biological stability where the shrinkage temperature and resistance to enzymatic degradation varied in response to ethanol/water solvent composition-mediated generation of cross-links. Irrespective of their cross-linking density, all studied membrane samples did not induce any responses in ocular epithelial cell cultures derived from cornea, lens, and retina. Unlike many other cross-linking agents and/or methods (e.g., excessive vapor phase cross-linking) that may pose a risk of toxicity, our study demonstrated that these nanofibrous materials are well tolerated by anterior segment tissues. These findings also indicate the safety of using ethanol/water co-solvents for chemical cross-linking of gelatin to engineer nanofibrous materials with negligible biological effects. In summary, the present results suggest the importance of solvent-mediated carbodiimide cross-linking in modulating structure�

Role of solvent-mediated carbodiimide cross-linking in fabrication of electrospun gelatin nanofibrous membranes as ophthalmic biomaterials

Energy Technology Data Exchange (ETDEWEB)

Chou, Shih-Feng [Department of Mechanical Engineering, University of Texas at Tyler, Tyler, TX 75799 (United States); Luo, Li-Jyuan [Department of Chemical and Materials Engineering, Chang Gung University, Taoyuan 33302, Taiwan, ROC (China); Lai, Jui-Yang, E-mail: jylai@mail.cgu.edu.tw [Institute of Biochemical and Biomedical Engineering, Chang Gung University, Taoyuan 33302, Taiwan, ROC (China); Biomedical Engineering Research Center, Chang Gung University, Taoyuan 33302, Taiwan, ROC (China); Center for Tissue Engineering, Chang Gung Memorial Hospital, Taoyuan 33305, Taiwan, ROC (China); Department of Ophthalmology, Chang Gung Memorial Hospital, Taoyuan 33305, Taiwan, ROC (China); Department of Materials Engineering, Ming Chi University of Technology, New Taipei City 24301, Taiwan, ROC (China); Ma, David Hui-Kang [Center for Tissue Engineering, Chang Gung Memorial Hospital, Taoyuan 33305, Taiwan, ROC (China); Department of Ophthalmology, Chang Gung Memorial Hospital, Taoyuan 33305, Taiwan, ROC (China); Department of Chinese Medicine, Chang Gung University, Taoyuan 33302, Taiwan, ROC (China)

2017-02-01

Due to their ability to mimic the structure of extracellular matrix, electrospun gelatin nanofibers are promising cell scaffolding materials for tissue engineering applications. However, the hydrophilic gelatin molecules usually need stabilization before use in aqueous physiological environment. Considering that biomaterials cross-linked via film immersion technique may have a more homogeneous cross-linked structure than vapor phase cross-linking, this work aims to investigate the chemical modification of electrospun gelatin nanofibrous membranes by liquid phase carbodiimide in the presence of ethanol/water co-solvents with varying ethanol concentrations ranging from 80 to 99.5 vol%. The results of characterization showed that increasing water content in the binary reaction solvent system increases the extent of cross-linking of gelatin nanofibers, but simultaneously promotes the effect of biopolymer swelling and distortion in fiber mat structure. As compared to non-cross-linked counterparts, carbodiimide treated gelatin nanofibrous mats exhibited better thermal and biological stability where the shrinkage temperature and resistance to enzymatic degradation varied in response to ethanol/water solvent composition-mediated generation of cross-links. Irrespective of their cross-linking density, all studied membrane samples did not induce any responses in ocular epithelial cell cultures derived from cornea, lens, and retina. Unlike many other cross-linking agents and/or methods (e.g., excessive vapor phase cross-linking) that may pose a risk of toxicity, our study demonstrated that these nanofibrous materials are well tolerated by anterior segment tissues. These findings also indicate the safety of using ethanol/water co-solvents for chemical cross-linking of gelatin to engineer nanofibrous materials with negligible biological effects. In summary, the present results suggest the importance of solvent-mediated carbodiimide cross-linking in modulating structure�

Bone matrix calcification during embryonic and postembryonic rat calvarial development assessed by SEM-EDX spectroscopy, XRD, and FTIR spectroscopy.

Science.gov (United States)

Henmi, Akiko; Okata, Hiroshi; Anada, Takahisa; Yoshinari, Mariko; Mikami, Yasuto; Suzuki, Osamu; Sasano, Yasuyuki

2016-01-01

Bone mineral is constituted of biological hydroxyapatite crystals. In developing bone, the mineral crystal matures and the Ca/P ratio increases. However, how an increase in the Ca/P ratio is involved in maturation of the crystal is not known. The relationships among organic components and mineral changes are also unclear. The study was designed to investigate the process of calcification during rat calvarial bone development. Calcification was evaluated by analyzing the atomic distribution and concentration of Ca, P, and C with scanning electron microscopy (SEM)-energy-dispersive X-ray (EDX) spectroscopy and changes in the crystal structure with X-ray diffraction (XRD) and Fourier transform infrared (FTIR) spectroscopy. Histological analysis showed that rat calvarial bone formation started around embryonic day 16. The areas of Ca and P expanded, matching the region of the developing bone matrix, whereas the area of C became localized around bone. X-ray diffraction and FTIR analysis showed that the amorphous-like structure of the minerals at embryonic day 16 gradually transformed into poorly crystalline hydroxyapatite, whereas the proportion of mineral to protein increased until postnatal week 6. FTIR analysis also showed that crystallization of hydroxyapatite started around embryonic day 20, by which time SEM-EDX spectroscopy showed that the Ca/P ratio had increased and the C/Ca and C/P ratios had decreased significantly. The study suggests that the Ca/P molar ratio increases and the proportion of organic components such as proteins of the bone matrix decreases during the early stage of calcification, whereas crystal maturation continues throughout embryonic and postembryonic bone development.

Altered composition of bone as triggered by irradiation facilitates the rapid erosion of the matrix by both cellular and physicochemical processes.

Directory of Open Access Journals (Sweden)

Danielle E Green

Full Text Available Radiation rapidly undermines trabecular architecture, a destructive process which proceeds despite a devastated cell population. In addition to the 'biologically orchestrated' resorption of the matrix by osteoclasts, physicochemical processes enabled by a damaged matrix may contribute to the rapid erosion of bone quality. 8w male C57BL/6 mice exposed to 5 Gy of Cs(137 γ-irradiation were compared to age-matched control at 2d, 10d, or 8w following exposure. By 10d, irradiation had led to significant loss of trabecular bone volume fraction. Assessed by reflection-based Fourier transform infrared imaging (FTIRI, chemical composition of the irradiated matrix indicated that mineralization had diminished at 2d by -4.3±4.8%, and at 10d by -5.8±3.2%. These data suggest that irradiation facilitates the dissolution of the matrix through a change in the material itself, a conclusion supported by a 13.7±4.5% increase in the elastic modulus as measured by nanoindentation. The decline in viable cells within the marrow of irradiated mice at 2d implies that the immediate collapse of bone quality and inherent increased risk of fracture is not solely a result of an overly-active biologic process, but one fostered by alterations in the material matrix that predisposes the material to erosion.

Solubilization of bovine gelatin using power ultrasound: gelation without heating.

Science.gov (United States)

Farahnaky, Asgar; Zendeboodi, Fatemeh; Azizi, Rezvan; Mesbahi, Gholamreza; Majzoobi, Mahsa

2017-04-01

The aim of this study was to investigate the efficacy of power ultrasound without using any heating stage in solubilizeing gelatin dispersions, and to characterize the mechanical and microstructural properties of the resulting gels using texture analysis and scanning electron microscopy, respectively. Usually to prepare a gel from gelatin, a primary heating stage of at about 40C or above is required to solubilize gelatin macromolecules. In this study solubilizing gelatin dispersions using power ultrasound without any heating was successfully performed. For solubilising gelatin, an ultrasound equipment with a frequency of 20 kHz, amplitude of 100% and power range of 50-150 W was used. Aqueous gelatin dispersions (4% w/v) were subjected to ultrasound for different times (40-240 s) at a constant temperature of 13C. Applying ultrasound to gelatin dispersions caused increases in water absorption and water solubility of the hydrocolloid. The textural parameters of the resulting gelatin gels, increased with increasing time and power of ultrasound. Moreover, a generalized Maxwell model with three elements was used for calculating relaxation times of the gels. The microstructural observations by SEM showed that the structural cohesiveness of the gels increased by increasing ultrasonication time. Ultrasound-assisted solubilization of gelatin can have emerging implications for industrial uses in pharmaceuticals, food and non-food systems. Usually to prepare a gel from gelatin, a primary heating stage of at about 40C or above is required to solubilize gelatin macromolecules. Therefore, the use of gelatin as a hydrocolloid in food processings or pharmaceutical formulations which lack a heating step has been a technological and practical challenge. In this study solubilizing gelatin dispersions using power ultrasound without any heating was successfully performed. Ultrasound-assisted solubilisation of gelatin can have emerging implications for industrial uses in pharmaceuticals

Preparation and characterization of hydroxyapatite/gelatin composite membranes for immunoisolation

Science.gov (United States)

Chen, Jyh-Ping; Chang, Feng-Nian

2012-12-01

Composite membranes are fabricated from hydroxyapatite (HAP) and gelatin for immunoisolation of cells. The films were fabricated by crosslinking 5 wt%, 10 wt%, and 20 wt% gelatin with 1 wt% glutaraldehyde (GA) in the presence of HAP. Fourier transform infrared spectroscopy analysis confirms imide bond formation between GA and gelatin, while the crystal structure of HAP powder remains unchanged from X-ray diffraction analysis. The degree of crosslinking depends on crosslinking time and gelatin concentration. For 5% and 10% gelatin, the degree of crosslinking levels off at 90% within 48 h. From scanning electron microscopy micrographs, the microstructure of the composite membrane depends on the amount of gelatin used in the crosslinking reaction. The mechanical strength of the composite membrane could be enhanced by increasing the gelatin concentration. BET analysis indicates that pore size of the micropores on the surface HAP/gelatin agglomerates decreases with increasing gelatin concentration. However, the macropore, through which diffusion of molecules occurs, is larger at higher gelatin concentrations. The permeability coefficients of different molecules through a HAP/gelatin composite membrane increase with increasing gelatin concentration and is inversely correlated with the molecular weight of the molecule. For immunoisolation of cells, the diffusion of large molecules stimulated by the immune system can be rejected by a chamber constructed from the HAP/gelatin membrane. Insulinoma cells were encapsulated in alginate-poly-L-lysine-alginate microcapsules and enclosed in a HAP/gelatin chamber. The chamber did not impair the viability and function of insulinoma cells and cells can secrete insulin in response to glucose concentration change. The chamber is therefore useful for the physiologically controlled secretion of insulin in response to the blood glucose level. Intraperitoneal transplantation of the chamber into streptozotocin-induced diabetic SD rats could

Bone Mineral 31P and Matrix-Bound Water Densities Measured by Solid-State 1H and 31P MRI

Science.gov (United States)

Seifert, Alan C.; Li, Cheng; Rajapakse, Chamith S.; Bashoor- Zadeh, Mahdieh; Bhagat, Yusuf A.; Wright, Alexander C.; Zemel, Babette S.; Zavaliangos, Antonios; Wehrli, Felix W.

2014-01-01

Bone is a composite material consisting of mineral and hydrated collagen fractions. MRI of bone is challenging due to extremely short transverse relaxation times, but solid-state imaging sequences exist that can acquire the short-lived signal from bone tissue. Previous work to quantify bone density via MRI used powerful experimental scanners. This work seeks to establish the feasibility of MRI-based measurement on clinical scanners of bone mineral and collagen-bound water densities, the latter as a surrogate of matrix density, and to examine the associations of these parameters with porosity and donors’ age. Mineral and matrix-bound water images of reference phantoms and cortical bone from 16 human donors, ages 27-97 years, were acquired by zero-echo-time 31P and 1H MRI on whole body 7T and 3T scanners, respectively. Images were corrected for relaxation and RF inhomogeneity to obtain density maps. Cortical porosity was measured by micro-CT, and apparent mineral density by pQCT. MRI-derived densities were compared to x-ray-based measurements by least-squares regression. Mean bone mineral 31P density was 6.74±1.22 mol/L (corresponding to 1129±204 mg/cc mineral), and mean bound water 1H density was 31.3±4.2 mol/L (corresponding to 28.3±3.7 %v/v). Both 31P and bound water (BW) densities were correlated negatively with porosity (31P: R2 = 0.32, p bone mineralization ratio (expressed here as the ratio of 31P density to bound water density), which is proportional to true bone mineralization, was found to be uncorrelated with porosity, age, or pQCT density. This work establishes the feasibility of image-based quantification of bone mineral and bound water densities using clinical hardware. PMID:24846186

[Bone Cell Biology Assessed by Microscopic Approach. Bone mineralization by ultrastructural imaging].

Science.gov (United States)

Hasegawa, Tomoka

2015-10-01

Bone mineralization can be divided into two phases ; one is primary mineralization associated with osteoblastic bone formation, and the other is secondary mineralization which gradually increases mineral density of bone matrix after the primary mineralization. Primary mineralization is initiated by matrix vesicles synthesized by mature osteoblasts. Crystalline calcium phosphates are nucleated inside these matrix vesicles, and then, get out of them forming spherical mineralized nodule, which can grow more by being supplied with Ca2+ and PO4(3-) (matrix vesicle mineralization). Thereafter, the mineralized nodules make contacts with surrounding collagen fibrils, extending mineralization along with their longitudinal axis from the contact points (collagen mineralization). In this review, the ultrastructural findings on bone mineralization, specially, primary mineralization will be provided.

Chest wall reconstruction in a canine model using polydioxanone mesh, demineralized bone matrix and bone marrow stromal cells.

Science.gov (United States)

Tang, Hua; Xu, Zhifei; Qin, Xiong; Wu, Bin; Wu, Lihui; Zhao, XueWei; Li, Yulin

2009-07-01

Extensive chest wall defect reconstruction remains a challenging problem for surgeons. In the past several years, little progress has been made in this area. In this study, a biodegradable polydioxanone (PDO) mesh and demineralized bone matrix (DBM) seeded with osteogenically induced bone marrow stromal cells (BMSCs) were used to reconstruct a 6 cm x 5.5 cm chest wall defect. Four experimental groups were evaluated (n=6 per group): polydioxanone (PDO) mesh/DBMs/BMSCs group, polydioxanone (PDO) mesh/DBMs group, polydioxanone (PDO) mesh group, and a blank group (no materials) in a canine model. All the animals survived except those in the blank group. In all groups receiving biomaterial implants, the polydioxanone (PDO) mesh completely degraded at 24 weeks and was replaced by fibrous tissue with thickness close to that of the normal intercostal tissue (P>0.05). In the polydioxanone (PDO) mesh/DBMs/BMSCs group, new bone formation and bone-union were observed by radiographic and histological examination. More importantly, the reconstructed rib could maintain its original radian and achieve satisfactory biomechanics close to normal ribs in terms of bending stress (P>0.05). However, in the other two groups, fibrous tissue was observed in the defect and junctions, and the reconstructed ribs were easily distorted under an outer force. Based on these results, a surgical approach utilizing biodegradable polydioxanone (PDO) mesh in combination with DBMs and BMSCs could repair the chest wall defect not only in function but also in structure.

Controlled Fabrication of Gelatin Nanoparticles as Drug Carriers

Science.gov (United States)

Jahanshahi, M.; Sanati, M. H.; Minuchehr, Z.; Hajizadeh, S.; Babaei, Z.

2007-08-01

In recent years, significant effort has been devoted to develop nanotechnology for drug delivery since it offers a suitable means of delivering small molecular weight drugs, as well as macromolecules such as proteins, peptides or genes by either localized or targeted delivery to the tissue of interest. Nanotechnology focuses on formulating therapeutic agents in biocompatible nanocomposites such as nanoparticles, nanocapsules, micellar systems, and conjugates. Protein nanoparticles (BSA, HAS and gelatin) generally vary in size from 50-300 nm and they hold certain advantages such as greater stability during storage, stability in vivo, non-toxicity, non-antigen and ease to scale up during manufacture over the other drug delivery systems. The primary structure of gelatin offers many possibilities for chemical modification and covalent drug attachment. Here nanoparticles of gelatin type A were prepared by a two-step desolvation method as a colloidal drug delivery system and the essential parameters in fabrication were considered. Gelatin was dissolved in 25 mL distilled water under room temperature range. Then acetone was added to the gelatin solution as a desolvating agent to precipitate the high molecular weight (HMW) gelatin. The supernatant was discarded and the HMW gelatin re-dissolved by adding 25 mL distilled water and stirring at 600 rpm. Acetone were added drop-wise to form nanoparticles. At the end of the process, glutaraldehyde solution was used for preparing nanoparticles as a cross-linking agent, and stirred for 12h at 600 rpm. For purification stage we use centrifuge with 600rpm for 3 times. The objective of the present study is consideration of some factors such as temperature, gelatin concentration, agitation speed and the amount of acetone and their effects on size and distribution of nanoparticles. Among the all conditions, 60° C, 50 mg/ml gelatin concentration, 75 ml acetone had the best result and the nanoparticle size was under 170 nm. The effect

Accurate 3-D Profile Extraction of Skull Bone Using an Ultrasound Matrix Array.

Science.gov (United States)

Hajian, Mehdi; Gaspar, Robert; Maev, Roman Gr

2017-12-01

The present study investigates the feasibility, accuracy, and precision of 3-D profile extraction of the human skull bone using a custom-designed ultrasound matrix transducer in Pulse-Echo. Due to the attenuative scattering properties of the skull, the backscattered echoes from the inner surface of the skull are severely degraded, attenuated, and at some points overlapped. Furthermore, the speed of sound (SOS) in the skull varies significantly in different zones and also from case to case; if considered constant, it introduces significant error to the profile measurement. A new method for simultaneous estimation of the skull profiles and the sound speed value is presented. The proposed method is a two-folded procedure: first, the arrival times of the backscattered echoes from the skull bone are estimated using multi-lag phase delay (MLPD) and modified space alternating generalized expectation maximization (SAGE) algorithms. Next, these arrival times are fed into an adaptive sound speed estimation algorithm to compute the optimal SOS value and subsequently, the skull bone thickness. For quantitative evaluation, the estimated bone phantom thicknesses were compared with the mechanical measurements. The accuracies of the bone thickness measurements using MLPD and modified SAGE algorithms combined with the adaptive SOS estimation were 7.93% and 4.21%, respectively. These values were 14.44% and 10.75% for the autocorrelation and cross-correlation methods. Additionally, the Bland-Altman plots showed the modified SAGE outperformed the other methods with -0.35 and 0.44 mm limits of agreement. No systematic error that could be related to the skull bone thickness was observed for this method.

Intervariability and intravariability of bone morphogenetic proteins in commercially available demineralized bone matrix products.

Science.gov (United States)

Bae, Hyun W; Zhao, Li; Kanim, Linda E A; Wong, Pamela; Delamarter, Rick B; Dawson, Edgar G

2006-05-20

Enzyme-linked immunosorbent assay was used to detect bone morphogenetic proteins (BMPs) 2, 4, and 7 in 9 commercially available ("off the shelf") demineralized bone matrix (DBM) product formulations using 3 different manufacturer's production lots of each DBM formulation. To evaluate and compare the quantity of BMPs among several different DBM formulations (inter-product variability), as well as examine the variability of these proteins in different production lots within the same DBM formulation (intra-product variability). DBMs are commonly used to augment available bone graft in spinal fusion procedures. Surgeons are presented with an ever-increasing variety of commercially available human DBMs from which to choose. Yet, there is limited information on a specific DBM product's osteoinductive efficacy, potency, and constancy. There were protein extracts from each DBM sample separately dialyzed 4 times against distilled water at 4 degrees C for 48 hours. The amount of BMP-2, BMP-4, and BMP-7 was determined using enzyme-linked immunosorbent assay. RESULTS.: The concentrations of detected BMP-2 and BMP-7 were low for all DBM formulations, only nanograms of BMP were extracted from each gram of DBM (20.2-120.6 ng BMP-2/g DBM product; 54.2-226.8 ng BMP-7/g DBM). The variability of BMP concentrations among different lots of the same DBM formulation, intra-product variability, was higher than the variability of concentrations among different DBM formulations, inter-product variability (coefficient of variation range BMP-2 [16.34% to 76.01%], P DBMs are low, in the order of 1 x 10(-9) g of BMP/g of DBM. There is higher variability in concentration of BMPs among 3 different lots of the same DBM formulation than among different DBM formulations. This variability questions DBM products' reliability and, possibly, efficacy in providing consistent osteoinduction.

Incorporation of mesoporous silica nanoparticles into random electrospun PLGA and PLGA/gelatin nanofibrous scaffolds enhances mechanical and cell proliferation properties

DEFF Research Database (Denmark)

Mehrasa, Mohammad; Asadollahi, Mohammad Ali; Nasri-Nasrabadi, Bijan

2016-01-01

Poly(lactic-co-glycolic.acid) (PLGA) and PLGA/gelatin random nanofibrous scaffolds embedded with different amounts of mesoporous silica nanoparticles (MSNPs) were fabricated using electrospinning method. To evaluate the effects of nanoparticles on the scaffolds, physical, chemical, and mechanical...... the porosity of scaffolds. Nanoparticles also improved the tensile mechanical properties of scaffolds. Using in vitro degradation analysis, it was shown that the addition of nanoparticles to the nano fibers matrix increases the weight loss percentage of PLGA-based samples, whereas it decreases the weight loss...... properties as well as in vitro degradation behavior of scaffolds were investigated. The mean diameters of nanofibers were 974 ± 68 nm for the pure PLGA scaffolds vs 832 ± 70, 764 ± 80, and 486 ± 64 for the PLGA/gelatin, PLGA/10 wt% MSNPs, and the PLGA/gelatin/10 wt% MSNPs scaffolds, respectively. The results...

Preparation and characterization of hydroxyapatite/gelatin composite membranes for immunoisolation

International Nuclear Information System (INIS)

Chen, Jyh-Ping; Chang, Feng-Nian

2012-01-01

Highlights: ► Cross-linking gelatin in the presence of hydroxyapatite forms composite membranes. ► The membrane was used for immunoisolation and encapsulation of cells. ► Encapsulated islet cells secrete insulin in response to glucose concentrations. ► The membrane is a good candidate for bioartificial pancreas development. - Abstract: Composite membranes are fabricated from hydroxyapatite (HAP) and gelatin for immunoisolation of cells. The films were fabricated by crosslinking 5 wt%, 10 wt%, and 20 wt% gelatin with 1 wt% glutaraldehyde (GA) in the presence of HAP. Fourier transform infrared spectroscopy analysis confirms imide bond formation between GA and gelatin, while the crystal structure of HAP powder remains unchanged from X-ray diffraction analysis. The degree of crosslinking depends on crosslinking time and gelatin concentration. For 5% and 10% gelatin, the degree of crosslinking levels off at 90% within 48 h. From scanning electron microscopy micrographs, the microstructure of the composite membrane depends on the amount of gelatin used in the crosslinking reaction. The mechanical strength of the composite membrane could be enhanced by increasing the gelatin concentration. BET analysis indicates that pore size of the micropores on the surface HAP/gelatin agglomerates decreases with increasing gelatin concentration. However, the macropore, through which diffusion of molecules occurs, is larger at higher gelatin concentrations. The permeability coefficients of different molecules through a HAP/gelatin composite membrane increase with increasing gelatin concentration and is inversely correlated with the molecular weight of the molecule. For immunoisolation of cells, the diffusion of large molecules stimulated by the immune system can be rejected by a chamber constructed from the HAP/gelatin membrane. Insulinoma cells were encapsulated in alginate-poly-L-lysine-alginate microcapsules and enclosed in a HAP/gelatin chamber. The chamber did not impair

Preparation and characterization of hydroxyapatite/gelatin composite membranes for immunoisolation

Energy Technology Data Exchange (ETDEWEB)

Chen, Jyh-Ping, E-mail: jpchen@mail.cgu.edu.tw [Department of Chemical and Materials Engineering, Chang Gung University, 259 Wen Hwa 1st Rd., Kwei-San, Taoyuan 333, Taiwan (China); Chang, Feng-Nian [Department of Chemical and Materials Engineering, Chang Gung University, 259 Wen Hwa 1st Rd., Kwei-San, Taoyuan 333, Taiwan (China)

2012-12-01

Highlights: Black-Right-Pointing-Pointer Cross-linking gelatin in the presence of hydroxyapatite forms composite membranes. Black-Right-Pointing-Pointer The membrane was used for immunoisolation and encapsulation of cells. Black-Right-Pointing-Pointer Encapsulated islet cells secrete insulin in response to glucose concentrations. Black-Right-Pointing-Pointer The membrane is a good candidate for bioartificial pancreas development. - Abstract: Composite membranes are fabricated from hydroxyapatite (HAP) and gelatin for immunoisolation of cells. The films were fabricated by crosslinking 5 wt%, 10 wt%, and 20 wt% gelatin with 1 wt% glutaraldehyde (GA) in the presence of HAP. Fourier transform infrared spectroscopy analysis confirms imide bond formation between GA and gelatin, while the crystal structure of HAP powder remains unchanged from X-ray diffraction analysis. The degree of crosslinking depends on crosslinking time and gelatin concentration. For 5% and 10% gelatin, the degree of crosslinking levels off at 90% within 48 h. From scanning electron microscopy micrographs, the microstructure of the composite membrane depends on the amount of gelatin used in the crosslinking reaction. The mechanical strength of the composite membrane could be enhanced by increasing the gelatin concentration. BET analysis indicates that pore size of the micropores on the surface HAP/gelatin agglomerates decreases with increasing gelatin concentration. However, the macropore, through which diffusion of molecules occurs, is larger at higher gelatin concentrations. The permeability coefficients of different molecules through a HAP/gelatin composite membrane increase with increasing gelatin concentration and is inversely correlated with the molecular weight of the molecule. For immunoisolation of cells, the diffusion of large molecules stimulated by the immune system can be rejected by a chamber constructed from the HAP/gelatin membrane. Insulinoma cells were encapsulated in alginate

Effect of dose rate on radical and property of gelatin

International Nuclear Information System (INIS)

Geng Shengrong; Chen Yuxia; Zu Xiaoyan; Li Xin; Jiang Hongyou

2015-01-01

The gelatin was irradiated respectively in the range of 0-32 kGy by dose rates of 60 Gy/min 60 Co, 480 Gy/min 60 Co and 12000 Gy/min accelerator, and the relationships of the radical character and gelatin property with dose rate were investigated through electron spin resonance (ESR) and gelatin permeation chromatogram. The results show that there is weak ESR signal from unirradiated gelatin, but irradiated one presents typical double peak. The order of ESR signal intensity of gelatin with the same absorbed dosage from high to low is 60 Gy/min 60 Co, 480 Gy/min 60 Co and 12000 Gy/min accelerator. The linear relationship between ESR signal intensity from 60 Co irradiated gelatin and absorbed dose is y= 26.983x 2 +1 641.8x-205.69. The intrinsic viscosity, average relative molecular weight, gelatin strength and breaking elongation of irradiated gelatin from high to low are 480 Gy/min 60 Co, 12000 Gy/min accelerator and 60 Gy/min 60 Co. The protection mechanism of high dose rate radiation on gelatin degradation is that the production of effective long life free radicals reduces. (authors)

Transplantation of artificial gelatin-co-bletillastriata gelatin/Salvia ...

African Journals Online (AJOL)

Tropical Journal of Pharmaceutical Research April 2016; 15 (4): 735-741 ... Methods: The composite artificial corium was constructed by culturing fibroblast cells in gelatin-co- ... Conclusion: The composite artificial corium has some clinical prospects for use in the treatment of ... burns, and treatment of large areas of skin.

Protective Effects of LSGYGP from Fish Skin Gelatin Hydrolysates on UVB-Induced MEFs by Regulation of Oxidative Stress and Matrix Metalloproteinase Activity.

Science.gov (United States)

Ma, Qingyu; Liu, Qiuming; Yuan, Ling; Zhuang, Yongliang

2018-03-28

A previous study has shown that tilapia fish skin gelatin hydrolysates inhibited photoaging in vivo, and that, Leu-Ser-Gly-Tyr-Gly-Pro (LSGYGP) identified in the hydrolysate had a high hydroxyl radical scavenging activity. In this study, activities of LSGYGP were further evaluated using ultraviolet B (UVB)-induced mouse embryonic fibroblasts (MEFs). UVB irradiation significantly increased the intercellular reactive oxygen species (ROS) production and matrix metalloproteinases (MMPs) activities and decreased the content of collagen in MEFs. LSGYGP reduced the intercellular ROS generation in UVB-induced MEFs. Meanwhile, the decrease of superoxide dismutase (SOD) activity and the increase of malondiaidehyde (MDA) content were inhibited by LSGYGP. LSGYGP reduced MMP-1 and MMP-9 activities in a dose-dependent manner. Molecular docking simulation indicated that LSGYGP inhibited MMPs activities by docking the active sites of MMP-1 and MMP-9. Furthermore, LSGYGP also affected the intercellular phosphorylation of UVB-induced the mitogen-activated protein kinase pathway. LSGYGP could protect collagen synthesis in MEFs under UVB irradiation by inhibiting oxidative stress and regulating MMPs activities.

Protective Effects of LSGYGP from Fish Skin Gelatin Hydrolysates on UVB-Induced MEFs by Regulation of Oxidative Stress and Matrix Metalloproteinase Activity

Directory of Open Access Journals (Sweden)

Qingyu Ma

2018-03-01

Full Text Available A previous study has shown that tilapia fish skin gelatin hydrolysates inhibited photoaging in vivo, and that, Leu-Ser-Gly-Tyr-Gly-Pro (LSGYGP identified in the hydrolysate had a high hydroxyl radical scavenging activity. In this study, activities of LSGYGP were further evaluated using ultraviolet B (UVB-induced mouse embryonic fibroblasts (MEFs. UVB irradiation significantly increased the intercellular reactive oxygen species (ROS production and matrix metalloproteinases (MMPs activities and decreased the content of collagen in MEFs. LSGYGP reduced the intercellular ROS generation in UVB-induced MEFs. Meanwhile, the decrease of superoxide dismutase (SOD activity and the increase of malondiaidehyde (MDA content were inhibited by LSGYGP. LSGYGP reduced MMP-1 and MMP-9 activities in a dose-dependent manner. Molecular docking simulation indicated that LSGYGP inhibited MMPs activities by docking the active sites of MMP-1 and MMP-9. Furthermore, LSGYGP also affected the intercellular phosphorylation of UVB-induced the mitogen-activated protein kinase pathway. LSGYGP could protect collagen synthesis in MEFs under UVB irradiation by inhibiting oxidative stress and regulating MMPs activities.

Sequential VEGF and BMP-2 releasing PLA-PEG-PLA scaffolds for bone tissue engineering: I. Design and in vitro tests.

Science.gov (United States)

Eğri, Sinan; Eczacıoğlu, Numan

2017-03-01

Biodegradable PLA-PEG-PLA block copolymers were synthesized with desired backbone structures and molecular weights using PEG20000. Rectangular scaffolds were prepared by freeze drying with or without using NaCl particles. Bone morphogenetic protein (BMP)-2 was loaded to the matrix after the scaffold formation for sustained release while vascular endothelial growth factor (VEGF) was loaded within the pores with gelatin solution. VEGF release was quite fast and almost 60% of it was released in 2 d. However, sequential - sustained released was observed for BMP-2 in the following few months. Corporation of VEGF/BMP-2 couple into the scaffolds increased the cell adhesion and proliferation. Neither significant cytotoxicity nor apoptosis/necrosis were observed.

Dentin matrix gelatin (DMG) as a possible "universal" grafting material in periodontics.

Science.gov (United States)

Gould, T R; Westbury, L; Tillman, J

1982-01-01

The ideal of periodontal surgery is the total regeneration of the lost periodontal complex. A promising new osseous grafting material is Dental Matrix Gelation (DMG). DMG was prepared by a method similar to that of Conover and Urist (1979). This consisted of sequential extraction in 1:1 chloroform-methanol, 25 degrees C for 1 hour; 0.6 N HCl, 2 degrees C for 24 hours with constant agitation; 2 M CaCl2, 2 degrees C for 1 hour; 0.5 M EDTA pH 7.4, 2 degrees C for 1 hour; washed in distilled water 1 hour. Twelve rats were anesthetized, had heads shaved, midline flaps reflected, and 2 mm holes drilled through the right and left parietal bones. This type of osseous defect normally heals only by fibrous scarring and has been used to define osteoinductive materials. The DMG was cut into pieces about 1 mm square and placed into the right side defect while the left side remained open as a control. The animals were sacrificed on a schedule of two rats every 2 weeks until the 10th week when four rats were killed. The results showed complete osseous closure of the DMG site while the control healed by fibrous scarring. DMG seems to have strong osteoinductive power, and used allogenically has great potential as a commercially viable implant material.

Irradiation of gelatin. Important applications for the development of new materials

Energy Technology Data Exchange (ETDEWEB)

Del Mastro, N. L., E-mail: nlmastro@ipen.br [Instituto de Pesquisas Energeticas e Nucleares IPEN-CNEN/SP, Av. Prof. Lineu Prestes 2242, 05008-900 Sao Paulo (Brazil)

2011-11-15

Gelatin is obtained from a naturally occurring protein, collagen, by chemical and thermal hydrolysis. As a protein, gelatin is biodegradable. Among biomaterials, gelatin is an interesting material because the dehydrated gelatin is a partially crystalline polymer and has a relatively low melting point. Gelatin melts to a liquid when heated and solidifies when cooled again. Together with water, it forms a semi-solid colloid gel. If gelatin is placed in contact with cold water, some of the material dissolves, their solubility being determined by their manufacturing method. Ionizing radiation acting on aqueous biological systems produces labile intermediates. Macromolecular free radicals may be diffusion ally mobile in homogenous solutions but relatively immobile in gelled systems. In this paper, different works, some of them performed in our laboratory, are firstly described showing gelatin irradiation in diverse systems. We have already studied the mechanical properties of gelatin composites prepared with a natural fiber, plasticizer and treated by electron beam irradiation. Departing from that, the range of novel applications for gelatin composites like gelatin nanoparticles as biodegradable s and low cell toxic alternative carrier delivery systems are outlined. The potential and the possibilities of using gelatin irradiation for important applications for the development of new materials for medical and food industry are presented. (Author)

The decontamination effects of gamma irradiation on the edible gelatin

International Nuclear Information System (INIS)

Fu, Junjie; Shen, Weiqiao; Bao, Jinsong; Chen, Qinglong

2000-01-01

The decontamination effects of gamma irradiation on the edible gelatin were studied. The results indicated that the bacterium and mold in the gelatin decreased significantly with the dose of 5 kGy treatment. However, the content of crude protein, microelement, amino acid in the gelatin remained unchanged under the irradiation of 4 and 8 kGy. The viscosity of the gelatin decreased with the increase of the irradiation dose, but the gelatin with a dose of 5 kGy treatment still accorded with the standard of the second-order class. These results suggested that the optimum irradiation dose for edible gelatin for the purpose of decontamination was in the range 3-5 kGy. (author)

Biomimetically-mineralized composite coatings on titanium functionalized with gelatin methacrylate hydrogels

Energy Technology Data Exchange (ETDEWEB)

Tan, Guoxin, E-mail: tanguoxin@126.com [School of Chemical Engineering and Light Industry, Guangdong University of Technology, Guangzhou, 510006 (China); Zhou, Lei [School of Chemical Engineering and Light Industry, Guangdong University of Technology, Guangzhou, 510006 (China); Ning, Chengyun, E-mail: imcyning@scut.edu.cn [College of Materials Science and Technology, South China University of Technology, Guangzhou, 510641 (China); Tan, Ying [School of Chemical Engineering and Light Industry, Guangdong University of Technology, Guangzhou, 510006 (China); Ni, Guoxin [Department of Orthopedics and Traumatology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515 (China); Liao, Jingwen; Yu, Peng; Chen, Xiaofeng [College of Materials Science and Technology, South China University of Technology, Guangzhou, 510641 (China)

2013-08-15

Immobilizing organic–inorganic hybrid composites onto the implant surface is a promising strategy to improve host acceptance of the implant. The objective of this present study was to obtain a unique macroporous titanium-surface with the organic–mineral composite coatings consisting of gelatin methacrylate hydrogel (GelMA) and hydroxyapatite (HA). A 3-(trimethoxysilyl) propyl methacrylate (TMSPMA) layer was first coated onto the titanium surface, and surface was then covalently functionalized with GelMA using a photochemical method. Mineralization of the GelMA coating on the titanium surface was subsequently carried out by a biomimetic method. After 3-day mineralization, a large number of mineral phases comprising spherical amorphous nanoparticles were found randomly deposited inside GelMA matrix. The resulting mineralized hydrogel composites exhibited a unique rough surface of macroporous structure. The structure of the prepared GelMA/HA composite coating was studied by field emission scanning electron microscopy (FESEM), energy dispersive X-ray spectra (EDS), attenuated total refraction Fourier transform infrared spectroscopy (ATR-FTIR), X-ray photoelectron spectroscopy (XPS) and X-ray diffraction (XRD). Water contact angle measurement revealed the hydrophilicity properties of composite coatings. GelMA/HA on titanium after the TMSPMA treatment is very stable when tested in vitro with a PBS solution at 37 °C, due to the role of TMSPMA as a molecular bridge. It was expected that the macroporous GelMA/HA composite coatings might potentially promote and accelerate titanium (Ti)-based implants osseointegration for bone repair and regeneration.

Fabrication and Characterization of Electrospun Polycaprolactone Blended with Chitosan-Gelatin Complex Nanofibrous Mats

Directory of Open Access Journals (Sweden)

Yongfang Qian

2014-01-01

Full Text Available Design and fabrication of nanofibrous scaffolds should mimic the native extracellular matrix. This study is aimed at investigating electrospinning of polycaprolactone (PCL blended with chitosan-gelatin complex. The morphologies were observed from scanning electron microscope. As-spun blended mats had thinner fibers than pure PCL. X-ray diffraction was used to analyze the degree of crystallinity. The intensity at two peaks at 2θ of 21° and 23.5° gradually decreased with the percentage of chitosan-gelatin complex increasing. Moreover, incorporation of the complex could obviously improve the hydrophilicity of as-spun blended mats. Mechanical properties of as-spun nanofibrous mats were also tested. The elongation at break of fibrous mats increased with the PCL content increasing and the ultimate tensile strength varied with different weight ratios. The as-spun mats had higher tensile strength when the weight ratio of PCL to CS-Gel was 75/25 compared to pure PCL. Both as-spun PCL scaffolds and PCL/CS-Gel scaffolds supported the proliferation of porcine iliac endothelial cells, and PCL/CS-Gel had better cell viability than pure PCL. Therefore, electrospun PCL/Chitosan-gelatin nanofibrous mats with weight ratio of 75/25 have better hydrophilicity mechanical properties, and cell proliferation and thus would be a promising candidate for tissue engineering scaffolds.

Biomimetic and enzyme-responsive dynamic hydrogels for studying cell-matrix interactions in pancreatic ductal adenocarcinoma.

Science.gov (United States)

Liu, Hung-Yi; Korc, Murray; Lin, Chien-Chi

2018-04-01

The tumor microenvironment (TME) governs all aspects of cancer progression and in vitro 3D cell culture platforms are increasingly developed to emulate the interactions between components of the stromal tissues and cancer cells. However, conventional cell culture platforms are inadequate in recapitulating the TME, which has complex compositions and dynamically changing matrix mechanics. In this study, we developed a dynamic gelatin-hyaluronic acid hybrid hydrogel system through integrating modular thiol-norbornene photopolymerization and enzyme-triggered on-demand matrix stiffening. In particular, gelatin was dually modified with norbornene and 4-hydroxyphenylacetic acid to render this bioactive protein photo-crosslinkable (through thiol-norbornene gelation) and responsive to tyrosinase-triggered on-demand stiffening (through HPA dimerization). In addition to the modified gelatin that provides basic cell adhesive motifs and protease cleavable sequences, hyaluronic acid (HA), an essential tumor matrix, was modularly and covalently incorporated into the cell-laden gel network. We systematically characterized macromer modification, gel crosslinking, as well as enzyme-triggered stiffening and degradation. We also evaluated the influence of matrix composition and dynamic stiffening on pancreatic ductal adenocarcinoma (PDAC) cell fate in 3D. We found that either HA-containing matrix or a dynamically stiffened microenvironment inhibited PDAC cell growth. Interestingly, these two factors synergistically induced cell phenotypic changes that resembled cell migration and/or invasion in 3D. Additional mRNA expression array analyses revealed changes unique to the presence of HA, to a stiffened microenvironment, or to the combination of both. Finally, we presented immunostaining and mRNA expression data to demonstrate that these irregular PDAC cell phenotypes were a result of matrix-induced epithelial-mesenchymal transition (EMT). Copyright © 2018 Elsevier Ltd. All rights

In vivo performance of novel soybean/gelatin-based bioactive and injectable hydroxyapatite foams.

Science.gov (United States)

Kovtun, Anna; Goeckelmann, Melanie J; Niclas, Antje A; Montufar, Edgar B; Ginebra, Maria-Pau; Planell, Josep A; Santin, Matteo; Ignatius, Anita

2015-01-01

Major limitations of calcium phosphate cements (CPCs) are their relatively slow degradation rate and the lack of macropores allowing the ingrowth of bone tissue. The development of self-setting cement foams has been proposed as a suitable strategy to overcome these limitations. In previous work we developed a gelatine-based hydroxyapatite foam (G-foam), which exhibited good injectability and cohesion, interconnected porosity and good biocompatibility in vitro. In the present study we evaluated the in vivo performance of the G-foam. Furthermore, we investigated whether enrichment of the foam with soybean extract (SG-foam) increased its bioactivity. G-foam, SG-foam and non-foamed CPC were implanted in a critical-size bone defect in the distal femoral condyle of New Zealand white rabbits. Bone formation and degradation of the materials were investigated after 4, 12 and 20weeks using histological and biomechanical methods. The foams maintained their macroporosity after injection and setting in vivo. Compared to non-foamed CPC, cellular degradation of the foams was considerably increased and accompanied by new bone formation. The additional functionalization with soybean extract in the SG-foam slightly reduced the degradation rate and positively influenced bone formation in the defect. Furthermore, both foams exhibited excellent biocompatibility, implying that these novel materials may be promising for clinical application in non-loaded bone defects. Copyright © 2014 Acta Materialia Inc. All rights reserved.

Dextran and gelatin based photocrosslinkable tissue adhesive.

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Wang, Tao; Nie, Jun; Yang, Dongzhi

2012-11-06

A two-component tissue adhesive based on biocompatible and bio-degradable polymers (oxidized urethane dextran (Dex-U-AD) and gelatin) was prepared and photocrosslinked under the ultraviolet (UV) irradiation. The adhesive could adhere to surface of gelatin, which simulated the human tissue steadily. The structures of above Dex-U-AD were characterized by FTIR, (1)H NMR spectroscopy and XRD. The adhesion property of result products was evaluated by lap-shear test. The maximum adhesion strength could reach to 4.16±0.72 MPa which was significantly higher than that of fibrin glue. The photopolymerization process of Dex-U-AD/gelatin was monitored by real time infrared spectroscopy (RTIR). It took less than 5 min to complete the curing process. The cytotoxicity of Dex-U-AD/gelatin also was evaluated which indicated that Dex-U-AD/gelatin gels were nontoxic to L929 cell. The relationship between all the above-mentioned properties and degree of oxidization of Dex-U-AD was assessed. The obtained products have the potential to serve as tissue adhesive in the future. Copyright © 2012 Elsevier Ltd. All rights reserved.

Organic composite-mediated surface coating of human acellular bone matrix with strontium.

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Huang, Yi-Zhou; Wang, Jing-Jing; Huang, Yong-Can; Wu, Cheng-Guang; Zhang, Yi; Zhang, Chao-Liang; Bai, Lin; Xie, Hui-Qi; Li, Zhao-Yang; Deng, Li

2018-03-01

Acellular bone matrix (ACBM) provides an osteoconductive scaffold for bone repair, but its osteoinductivity is poor. Strontium (Sr) improves the osteoinductivity of bone implants. In this study, we developed an organic composite-mediated strontium coating strategy for ACBM scaffolds by using the ion chelating ability of carboxymethyl cellulose (CMC) and the surface adhesion ability of dopamine (DOPA). The organic coating composite, termed the CMC-DOPA-Sr composite, was synthesized under a mild condition, and its chemical structure and strontium ion chelating ability were then determined. After surface decoration, the physicochemical properties of the strontium-coated ACBM (ACBM-Sr) scaffolds were characterized, and their biocompatibility and osteoinductivity were determined in vitro and in vivo. The results showed that the CMC-DOPA-Sr composite facilitated strontium coating on the surface of ACBM scaffolds. The ACBM-Sr scaffolds possessed a sustained strontium ion release profile, exhibited good cytocompatibility, and enhanced the osteogenic differentiation of mesenchymal stem cells in vitro. Furthermore, the ACBM-Sr scaffolds showed good histocompatibility after subcutaneous implantation in nude mice. Taken together, this study provided a simple and mild strategy to realize strontium coating for ACBM scaffolds, which resulted in good biocompatibility and improved osteoinductivity. Copyright © 2017 Elsevier B.V. All rights reserved.

[Children, Collect Bones! : Teaching Aids and Propaganda Material on Bone-Collections and Bone-Utilisation Used in German Schools During the "Third Reich"].

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Vaupel, Elisabeth; Preiß, Florian

2018-06-05

In the nineteenth and early twentieth centuries bones were an essential raw material for the German chemical industry, vital to the production of fertilizer, glue, gelatine, soap and other products. As most of this material was imported, the German school system during the "Third Reich" took the utilisation of bones as an example to illustrate the relevance of the four-year plan of 1936 and its policy of economic self-sufficiency. The school children were encouraged to collect bones from domestic sources and bring them to the collecting points in the schools. Several NS-institutions developed a variety of teaching aids and materials to support school education on this economically and politically important topic. Focussing on the example of bone-utilisation, this paper examines the messages and intentions of these educational materials. It also demonstrates how even apparently ideologically unbiased school subjects, such as chemistry, were instrumentalised for the political indoctrination of the pupils.

Characterization of Electrospun Nanofibrous Scaffolds for Nanobiomedical Applications

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Emul, E.; Saglam, S.; Ates, H.; Korkusuz, F.; Saglam, N.

2016-08-01

The electrospinning method is employed in the production of porous fiber scaffolds, and the usage of electrospun scaffolds especially as drug carrier and bone reconstructive material such as implants is promising for future applications in tissue engineering. The number of publications has grown very rapidly in this field through the fabrication of complex scaffolds, novel approaches in nanotechnology, and improvements of imaging methods. Hence, characterization of these materials has also grown significantly important for getting satisfied and accurate results. This advantageous and versatile method is ideal for mimicking bone extracellular matrix, and many biodegradable and biocompatible polymers are preferred in the field of bone reconstruction. In this study, gelatin, gelatin/nanohydroxyapatite (nHAp) and gelatin/PLLA/nHAp scaffolds were fabricated by the electrospinning process. These composite fibers showed clear and continuous morphology according to observation through a scanning electron microscope and their component analyses were also determined by Fourier transform infrared spectrometer analyses. These characterization experiments revealed the great effects of the electrospinning method for biomedical applications and have an especially important role in bone reconstruction and production of implant coating material.

Antimicrobial gelatin-based elastomer nanocomposite membrane loaded with ciprofloxacin and polymyxin B sulfate in halloysite nanotubes for wound dressing.

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Shi, Rui; Niu, Yuzhao; Gong, Min; Ye, Jingjing; Tian, Wei; Zhang, Liqun

2018-06-01

Bacterial infection is a major problem world-wide, especially in wound treatment where it can severely prolong the healing process. In this study, a double drug co-delivery elastic antibacterial nanocomposite was developed by combining ciprofloxacin (CPX) and polymyxin B sulfate-loaded halloysite clay nanotubes (HNTs-B) into a gelatin elastomer. CPX nanoparticles which act against both gram positive and gram-negative bacterium were dispersed directly in the matrix, and polymyxin B sulfate was loaded in HNTs and then distributed into the matrix. The effect of CPX and HNTs-B content on the physical properties, cytotoxicity, fibroblast adhesion and proliferation, in vitro drug release behavior and anti-bacterial properties were systematically investigated. The ciprofloxacin crystals and HNT-B were distributed in the matrix uniformly. The HNTs in the drug loading system not only enhanced the matrix' tensile strength but also slowed down the release rate of the high dissoluble polymyxin B sulfate. When the amount of HNT in the matrix increased, the thermal stability and tensile strength also increased but the polymyxin B sulfate release rate decreased because the HNTs prevented the drug release inside. All the nanocomposites exhibited antimicrobial activity against both gram-negative and gram-positive bacteria with the dual combination of drugs released from the nanocomposites. Furthermore, this kind of gelatin-based nanocomposites possesses higher water-absorbing quality, low cytotoxicity, adaptable biodegradability and good elasticity which can satisfy the requirements for an ideal biomaterial for use in wound healing applications. Copyright © 2018. Published by Elsevier B.V.

Gelatin Nano-coating for Inhibiting Surface Crystallization of Amorphous Drugs.

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Teerakapibal, Rattavut; Gui, Yue; Yu, Lian

2018-01-05

Inhibit the fast surface crystallization of amorphous drugs with gelatin nano-coatings. The free surface of amorphous films of indomethacin or nifedipine was coated by a gelatin solution (type A or B) and dried. The coating's effect on surface crystallization was evaluated. Coating thickness was estimated from mass change after coating. For indomethacin (weak acid, pK a  = 4.5), a gelatin coating of either type deposited at pH 5 and 10 inhibited its fast surface crystal growth. The coating thickness was 20 ± 10 nm. A gelatin coating deposited at pH 3, however, provided no protective effect. These results suggest that an effective gelatin coating does not require that the drug and the polymer have opposite charges. The ineffective pH 3 coating might reflect the poor wetting of indomethacin's neutral, hydrophobic surface by the coating solution. For nifedipine (weak base, pK a  = 2.6), a gelatin coating of either type deposited at pH 5 inhibited its fast surface crystal growth. Gelatin nano-coatings can be conveniently applied to amorphous drugs from solution to inhibit fast surface crystallization. Unlike strong polyelectrolyte coatings, a protective gelatin coating does not require strict pairing of opposite charges. This could make gelatin coating a versatile, pharmaceutically acceptable coating for stabilizing amorphous drugs.

Bone preparation at the KCCAMS laboratory

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Beaumont, Will, E-mail: wbeaumon@uci.ed [Earth System Science Dept., University of California, B321 Croul Hall, Irvine, CA 92697-3100 (United States); Beverly, Robert; Southon, John [Earth System Science Dept., University of California, B321 Croul Hall, Irvine, CA 92697-3100 (United States); Taylor, R.E. [Department of Anthropology, University of California, Riverside, CA 92521 (United States)

2010-04-15

Collagen purification by ultrafiltration is becoming widely accepted as a superior {sup 14}C sample preparation method for removing environmental contamination from archaeological bone. We present an overview of bone sample preparation methods used at the KCCAMS laboratory, UC Irvine. Methods development data are also discussed, including results of tests of gelatinization temperature, evaluation of 10 kDa vs. 30 kDa molecular weight (MW) ultrafiltration, improvements to ultrafilter cleaning methods, tests of ultrafilter memory, and ultrafilter glycerin contamination. These tests were not intended to be exhaustive, but rather to show how standard procedures can be validated and improved with relatively little effort.

The Feasibility of Gelatin-Based Retronasal Stimuli to Assess Olfactory Perception

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Daniel Shepherd

2015-10-01

Full Text Available Links between some psychological disorders and olfactory deficits are well documented, and screening tests have been developed to exploit these associations. Odors can take one of two routes to the olfactory receptors in the nasal epithelium, the orthonasal or retronasal route. This article discusses the potential use of the retronasal route to assess olfaction using gelatin-based stimuli delivered orally. Using a relatively new psychophysical method, the Single-Interval Adjustment Matrix task, we estimated vanillin thresholds for five healthy participants sampling small vanillin flavored gels. Our data demonstrate the feasibility of using solid-state gustatory stimuli to assess retronasal perception.

[Biomimetic nanohydroxyapatite/gelatin composite material preparation and in vitro study].

Science.gov (United States)

Li, Siriguleng; Hu, Xiaowen

2014-09-01

To prepare nHA/gelatin porous scaffold and to evaluate its physical and chemical properties and biocompatibility. We used nano-powders of HA and gelatin to prepare 3D porous composite scaffold by freeze-drying technique, and used scanning electron microscope, fourier transform infrared spectroscopy and universal testing machine to characterize the composite material. Osteoblasts were primarily cultured, and the third-passage osteoblasts were co-cultured with the composite material. The cell adhesion and morphology were examined under scanning electron microscope. The cell viability analysis was performed by MTT assay, and the alkaline phosphatase activity was measured with alkaline phosphatase kit. Scanning electron microscope showed that the scaffold possessed a 3-dimensional interconnected homogenous porous structure with pore sizes ranging from 150 to 400 μm. Fourier transform infrared spectroscopy showed that the composite material had a strong chemical bond between the inorganic phase and organic phase. The scaffold presented the compressive strength of (3.28 ± 0.51) MPa and porosities of (80.6 ± 4.1)%. Composite materials showed features of had good biocompatibility. Mouse osteoblasts were well adhered and spread on the materials. The grade of the cell toxicity ranged from I to II. On the 5th and 7th day the proliferative rate of osteoblasts on scaffolds in the composite materials was significantly higher than that in the control group. The activity of alkaline phosphatase was obviously higher than that in the control group on Day 1 and 3. Nano-hydroxyapatite and gelatin in certain proportions and under certain conditions can be prepared into a composite biomimetic porous scaffolds with high porosity and three-dimensional structure using freeze-drying method. The scaffold shows good biocompatibility with mouse osteoblasts and may be a novel scaffolds for bone tissue engineering.

Optical limiting in gelatin stabilized Cu-PVP nanocomposite colloidal suspension

Science.gov (United States)

Tamgadge, Y. S.; Gedam, P. P.; Thakare, N. B.; Talwatkar, S. S.; Sunatkari, A. L.; Muley, G. G.

2018-05-01

This article illustrates investigations on optical limiting properties of Cu-PVP nanocomposite colloidal suspension. Gelatin stabilized Cu nanoparticles have been synthesized using chemical reduction method and thin films in PVP matrix have been obtained using spin coating technique. Thin films have been characterized by X-ray diffraction (XRD), Ultraviolet-visible (UV-vis) spectroscopy, etc. for structural and linear optical studies. Optical limiting properties of Colloidal Cu-PVP nanocomposites have been investigated at 808 nm diode CW laser. Minimum optical limiting threshold was found for GCu3-PVP nanocomposites sample. The strong optical limiting is thermal in origin as CW laser is used and effects are attributed to thermal lensing effect.

Composite porous scaffold of PEG/PLA support improved bone matrix deposition in vitro compared to PLA-only scaffolds.

Science.gov (United States)

Bhaskar, Birru; Owen, Robert; Bahmaee, Hossein; Wally, Zena; Sreenivasa Rao, Parcha; Reilly, Gwendolen C

2018-05-01

Controllable pore size and architecture are essential properties for tissue-engineering scaffolds to support cell ingrowth colonization. To investigate the effect of polyethylene glycol (PEG) addition on porosity and bone-cell behavior, porous polylactic acid (PLA)-PEG scaffolds were developed with varied weight ratios of PLA-PEG (100/0, 90/10, 75/25) using solvent casting and porogen leaching. Sugar 200-300 µm in size was used as a porogen. To assess scaffold suitability for bone tissue engineering, MLO-A5 murine osteoblast cells were cultured and cell metabolic activity, alkaline phosphatase (ALP) activity and bone-matrix production determined using (alizarin red S staining for calcium and direct red 80 staining for collagen). It was found that metabolic activity was significantly higher over time on scaffolds containing PEG, ALP activity and mineralized matrix production were also significantly higher on scaffolds containing 25% PEG. Porous architecture and cell distribution and penetration into the scaffold were analyzed using SEM and confocal microscopy, revealing that inclusion of PEG increased pore interconnectivity and therefore cell ingrowth in comparison to pure PLA scaffolds. The results of this study confirmed that PLA-PEG porous scaffolds support mineralizing osteoblasts better than pure PLA scaffolds, indicating they have a high potential for use in bone tissue engineering applications. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1334-1340, 2018. © 2018 Wiley Periodicals, Inc.

Porous surface modified bioactive bone cement for enhanced bone bonding.

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Qiang He

Full Text Available Polymethylmethacrylate bone cement cannot provide an adhesive chemical bonding to form a stable cement-bone interface. Bioactive bone cements show bone bonding ability, but their clinical application is limited because bone resorption is observed after implantation. Porous polymethylmethacrylate can be achieved with the addition of carboxymethylcellulose, alginate and gelatin microparticles to promote bone ingrowth, but the mechanical properties are too low to be used in orthopedic applications. Bone ingrowth into cement could decrease the possibility of bone resorption and promote the formation of a stable interface. However, scarce literature is reported on bioactive bone cements that allow bone ingrowth. In this paper, we reported a porous surface modified bioactive bone cement with desired mechanical properties, which could allow for bone ingrowth.The porous surface modified bioactive bone cement was evaluated to determine its handling characteristics, mechanical properties and behavior in a simulated body fluid. The in vitro cellular responses of the samples were also investigated in terms of cell attachment, proliferation, and osteoblastic differentiation. Furthermore, bone ingrowth was examined in a rabbit femoral condyle defect model by using micro-CT imaging and histological analysis. The strength of the implant-bone interface was also investigated by push-out tests.The modified bone cement with a low content of bioactive fillers resulted in proper handling characteristics and adequate mechanical properties, but slightly affected its bioactivity. Moreover, the degree of attachment, proliferation and osteogenic differentiation of preosteoblast cells was also increased. The results of the push-out test revealed that higher interfacial bonding strength was achieved with the modified bone cement because of the formation of the apatite layer and the osseointegration after implantation in the bony defect.Our findings suggested a new bioactive

Gelatin-Based Hydrogels for Organ 3D Bioprinting

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Xiaohong Wang

2017-08-01

Full Text Available Three-dimensional (3D bioprinting is a family of enabling technologies that can be used to manufacture human organs with predefined hierarchical structures, material constituents and physiological functions. The main objective of these technologies is to produce high-throughput and/or customized organ substitutes (or bioartificial organs with heterogeneous cell types or stem cells along with other biomaterials that are able to repair, replace or restore the defect/failure counterparts. Gelatin-based hydrogels, such as gelatin/fibrinogen, gelatin/hyaluronan and gelatin/alginate/fibrinogen, have unique features in organ 3D bioprinting technologies. This article is an overview of the intrinsic/extrinsic properties of the gelatin-based hydrogels in organ 3D bioprinting areas with advanced technologies, theories and principles. The state of the art of the physical/chemical crosslinking methods of the gelatin-based hydrogels being used to overcome the weak mechanical properties is highlighted. A multicellular model made from adipose-derived stem cell proliferation and differentiation in the predefined 3D constructs is emphasized. Multi-nozzle extrusion-based organ 3D bioprinting technologies have the distinguished potential to eventually manufacture implantable bioartificial organs for purposes such as customized organ restoration, high-throughput drug screening and metabolic syndrome model establishment.

Advanced bredigite-containing magnesium-matrix composites for biodegradable bone implant applications.

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Dezfuli, Sina Naddaf; Huan, Zhiguang; Mol, Arjan; Leeflang, Sander; Chang, Jiang; Zhou, Jie

2017-10-01

The present research was aimed at developing magnesium-matrix composites that could allow effective control over their physiochemical and mechanical responses when in contact with physiological solutions. A biodegradable, bioactive ceramic - bredigite was chosen as the reinforcing phase in the composites, based on the hypothesis that the silicon- and magnesium-containing ceramic could protect magnesium from fast corrosion and at the same time stimulate cell proliferation. Methods to prepare composites with integrated microstructures - a prerequisite to achieve controlled biodegradation were developed. A systematic experimental approach was taken in order to elucidate the in vitro biodegradation mechanisms and kinetics of the composites. It was found that the composites with 20-40% homogenously dispersed bredigite particles, prepared from powders, could indeed significantly decrease the degradation rate of magnesium by up to 24 times. Slow degradation of the composites resulted in the retention of the mechanical integrity of the composites within the strength range of cortical bone after 12days of immersion in a cell culture medium. Cell attachment, cytotoxicity and bioactivity tests confirmed the stimulatory effects of bredigite embedded in the composites on the attachment, viability and differentiation of bone marrow stromal cells. Thus, the multiple benefits of adding bredigite to magnesium in enhancing degradation behavior, mechanical properties, biocompatibility and bioactivity were obtained. The results from this research showed the excellent potential of the bredigite-containing composites for bone implant applications, thus warranting further in vitro and in vivo research. Copyright © 2017 Elsevier B.V. All rights reserved.

Projectile penetration into ballistic gelatin.

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Swain, M V; Kieser, D C; Shah, S; Kieser, J A

2014-01-01

Ballistic gelatin is frequently used as a model for soft biological tissues that experience projectile impact. In this paper we investigate the response of a number of gelatin materials to the penetration of spherical steel projectiles (7 to 11mm diameter) with a range of lower impacting velocities (projectile velocity are found to be linear for all systems above a certain threshold velocity required for initiating penetration. The data for a specific material impacted with different diameter spheres were able to be condensed to a single curve when the penetration depth was normalised by the projectile diameter. When the results are compared with a number of predictive relationships available in the literature, it is found that over the range of projectiles and compositions used, the results fit a simple relationship that takes into account the projectile diameter, the threshold velocity for penetration into the gelatin and a value of the shear modulus of the gelatin estimated from the threshold velocity for penetration. The normalised depth is found to fit the elastic Froude number when this is modified to allow for a threshold impact velocity. The normalised penetration data are found to best fit this modified elastic Froude number with a slope of 1/2 instead of 1/3 as suggested by Akers and Belmonte (2006). Possible explanations for this difference are discussed. © 2013 Published by Elsevier Ltd.

Microencapsulation of sulforaphane from broccoli seed extracts by gelatin/gum arabic and gelatin/pectin complexes.

Science.gov (United States)

García-Saldaña, Jesús S; Campas-Baypoli, Olga N; López-Cervantes, Jaime; Sánchez-Machado, Dalia I; Cantú-Soto, Ernesto U; Rodríguez-Ramírez, Roberto

2016-06-15

Sulforaphane is a phytochemical that has received attention in recent years due to its chemopreventive properties. However, the uses and applications of this compound are very limited, because is an unstable molecule that is degraded mainly by changes in temperature and pH. In this research, the use of food grade polymers for microencapsulation of sulforaphane was studied by a complex coacervation method using the interaction of oppositely charged polymers as gelatin/gum arabic and gelatin/pectin. The polymers used were previously characterized in moisture content, ash and nitrogen. The encapsulation yield was over 80%. The gelatin/pectin complex had highest encapsulation efficiency with 17.91%. The presence of sulforaphane in the complexes was confirmed by FTIR and UV/visible spectroscopy. The materials used in this work could be a new and attractive option for the protection of sulforaphane. Copyright © 2016 Elsevier Ltd. All rights reserved.

Comparison of ballistic impact effects between biological tissue and gelatin.

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Jin, Yongxi; Mai, Ruimin; Wu, Cheng; Han, Ruiguo; Li, Bingcang

2018-02-01

Gelatin is commonly used in ballistic testing as substitute for biological tissue. Comparison of ballistic impact effects produced in the gelatin and living tissue is lacking. The work in this paper was aimed to compare the typical ballistic impact effects (penetration trajectory, energy transfer, temporary cavity) caused by 4.8mm steel ball penetrating the 60kg porcine hind limbs and 10wt% gelatin. The impact event in the biological tissue was recorded by high speed flash X-ray machine at different delay time, while the event in the gelatin continuously recorded by high speed video was compared to that in the biological tissue. The collected results clearly displayed that the ballistic impact effects in the muscle and gelatin were similar for the steel ball test; as for instance, the projectile trajectory in the two targets was basically similar, the process of energy transfer was highly coincident, and the expansion of temporary cavity followed the same pattern. This study fully demonstrated that choosing gelatin as muscle simulant was reasonable. However, the maximum temporary cavity diameter in the gelatin was a little larger than that in the muscle, and the expansion period of temporary cavity was longer in the gelatin. Additionally, the temporary cavity collapse process in the two targets followed different patterns, and the collapse period in the gelatin was two times as long as that in the muscle. Copyright © 2017 Elsevier Ltd. All rights reserved.

Regulation of extracellular matrix vesicles via rapid responses to steroid hormones during endochondral bone formation.

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Asmussen, Niels; Lin, Zhao; McClure, Michael J; Schwartz, Zvi; Boyan, Barbara D

2017-12-09

Endochondral bone formation is a precise and highly ordered process whose exact regulatory framework is still being elucidated. Multiple regulatory pathways are known to be involved. In some cases, regulation impacts gene expression, resulting in changes in chondrocyte phenotypic expression and extracellular matrix synthesis. Rapid regulatory mechanisms are also involved, resulting in release of enzymes, factors and micro RNAs stored in extracellular matrisomes called matrix vesicles. Vitamin D metabolites modulate endochondral development via both genomic and rapid membrane-associated signaling pathways. 1α,25-dihydroxyvitamin D3 [1α,25(OH) 2 D 3 ] acts through the vitamin D receptor (VDR) and a membrane associated receptor, protein disulfide isomerase A3 (PDIA3). 24R,25-dihydroxyvitamin D3 [24R,25(OH) 2 D 3 ] affects primarily chondrocytes in the resting zone (RC) of the growth plate, whereas 1α,25(OH) 2 D 3 affects cells in the prehypertrophic and upper hypertrophic cell zones (GC). This includes genomically directing the cells to produce matrix vesicles with zone specific characteristics. In addition, vitamin D metabolites produced by the cells interact directly with the matrix vesicle membrane via rapid signal transduction pathways, modulating their activity in the matrix. The matrix vesicle payload is able to rapidly impact the extracellular matrix via matrix processing enzymes as well as providing a feedback mechanism to the cells themselves via the contained micro RNAs. Copyright © 2017. Published by Elsevier Inc.

Preparation of gelatin-assisted polypyrrole-poly(3,4-ethylenedioxythiophene) composites

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Jo, Eun Jin; Male, Umashankar; Huh, Do Sung [Dept.of Chemistry and Nanosystem Engineering, Center for Nano Manufacturing, Inje University, Kimhae (Korea, Republic of)

2016-11-15

Conductive polymer composite was synthesized by the polymerization of pyrrole with 3,4-ethylenedioxythiophene using ammonium persulfate as an oxidant with different concentrations of gelatin dispersed in aqueous medium along with the polymerization. Differently from pure polypyrrole–poly(3,4-ethylenedioxythiophene) (PPy–PEDOT) copolymer, the gelatin-assisted composites showed good solubility in aqueous media. The prepared PPy–PEDOT/gelatin composites were identified with Fourier-transform infrared, UV–visible spectroscopy, and X-ray diffraction. Morphological analysis and temperature-dependent direct current conductivity of PPy–PEDOT/gelatin composites were carried out. Obtained results revealed that added gelatin greatly increased conductivity due to the improved mobility of charge carriers.

Drug loaded homogeneous electrospun PCL/gelatin hybrid nanofiber structures for anti-infective tissue regeneration membranes.

Science.gov (United States)

Xue, Jiajia; He, Min; Liu, Hao; Niu, Yuzhao; Crawford, Aileen; Coates, Phil D; Chen, Dafu; Shi, Rui; Zhang, Liqun

2014-11-01

Infection is the major reason for guided tissue regeneration/guided bone regeneration (GTR/GBR) membrane failure in clinical application. In this work, we developed GTR/GBR membranes with localized drug delivery function to prevent infection by electrospinning of poly(ε-caprolactone) (PCL) and gelatin blended with metronidazole (MNA). Acetic acid (HAc) was introduced to improve the miscibility of PCL and gelatin to fabricate homogeneous hybrid nanofiber membranes. The effects of the addition of HAc and the MNA content (0, 1, 5, 10, 20, 30, and 40 wt.% of polymer) on the properties of the membranes were investigated. The membranes showed good mechanical properties, appropriate biodegradation rate and barrier function. The controlled and sustained release of MNA from the membranes significantly prevented the colonization of anaerobic bacteria. Cells could adhere to and proliferate on the membranes without cytotoxicity until the MNA content reached 30%. Subcutaneous implantation in rabbits for 8 months demonstrated that MNA-loaded membranes evoked a less severe inflammatory response depending on the dose of MNA than bare membranes. The biodegradation time of the membranes was appropriate for tissue regeneration. These results indicated the potential for using MNA-loaded PCL/gelatin electrospun membranes as anti-infective GTR/GBR membranes to optimize clinical application of GTR/GBR strategies. Copyright © 2014 Elsevier Ltd. All rights reserved.

In vitro bone formation using muscle-derived cells: a new paradigm for bone tissue engineering using polymer-bone morphogenetic protein matrices.

Science.gov (United States)

Lu, Helen H; Kofron, Michelle D; El-Amin, Saadiq F; Attawia, Mohammed A; Laurencin, Cato T

2003-06-13

Over 800,000 bone grafting procedures are performed in the United States annually, creating a demand for viable alternatives to autogenous bone, the grafting standard in osseous repair. The objective of this study was to examine the efficacy of a BMP-polymer matrix in inducing the expression of the osteoblastic phenotype and in vitro bone formation by muscle-derived cells. Specifically, we evaluated the ability of bone morphogenetic protein-7 (BMP-7), delivered from a poly(lactide-co-glycolide) (PLAGA) matrix, to induce the differentiation of cells derived from rabbit skeletal muscle into osteoblast-like cells and subsequently form mineralized tissue. Results confirmed that muscle-derived cells attached and proliferated on the PLAGA substrates. BMP-7 released from PLAGA induced the muscle-derived cells to increase bone marker expression and form mineralized cultures. These results demonstrate the efficacy of a BMP-polymer matrix in inducing the expression of the osteoblastic phenotype by muscle-derived cells and present a new paradigm for bone tissue engineering.

Advances in allogenic bone graft processing and usage: preparation and evaluation of chitosan-demineralized cancellous bone powder composite scaffolds as a bone graft substitute

International Nuclear Information System (INIS)

Yongyudh Vajaradul

2008-01-01

Full text: Demineralized bone matrix (DBM) is currently used by surgeons. It usually exists as a lyophilized powder which is difficult to handle and operated. In this study, we try to improve these disadvantages by combining DBM with a biomaterial. It focuses on a natural biodegradable polymer, chitosan, to act as a temporary matrix for bone growth that easily prepare in any size and shape by using tissue engineering knowledge to get a proper temporary matrix. Thus, the development of chitosan-demineralized bone powder composite scaffold is an alternative way. Polymeric scaffold has been demonstrated to have great potential for tissue engineering because the scaffold or three dimension (3D) construct provides the necessary support for cells to proliferate, extracellular matrix deposition and vascularization of neo-tissue. Moreover, chitosan, a natural cationic polymer which its structural is similar to extracellular matrix glycosaminoblycans, is biodegradable, biocompatible, non-antigenic and biofunctional. It can enhance osteoblast cells proliferation and mineral matrix deposition in culture. The first study was to fabricate and analyze composite scaffold composed of either chitosan-demineralized cancellous bone powders or chitosan-demineralized cancellous cartilage bone powders in a ratio 50:50 and 70:30 w/w (chitosan : bone powders) based on physical properties composing of average pore diameter, mechanical integrity and swelling property. Secondly, scaffolds were evaluated in term of biological properties composing of their ability to support neo osteogenesis, including assessments of cell attachment and viability, cell morphology, and the biosynthesis of extracellular matrix. Results indicated that chitosan-demineralized cancellous bone powder composite scaffolds possessing an interconnecting, porous structure could be easily created through a simple freezing and lyophilization process. (Author)

Synthesis and Property of Ag(NP)/catechin/Gelatin Nanofiber

Science.gov (United States)

Nasir, Muhamad; Apriani, Dita

2017-12-01

Nanomaterial play important role future industry such as for the medical, food, pharmaceutical and cosmetic industry. Ag (NP) and catechin exhibit antibacterial property. Ag(NP) with diameter around 15 nm was synthesis by microwaved method. We have successfully produce Ag(NP)/catechin/gelatin nanofiber composite by electrospinning process. Ag(NP)/catechin/gelatin nanofiber was synthesized by using gelatin from tuna fish, polyethylene oxide (PEO), acetic acid as solvent and silver nanoparticle(NP)/catechin as bioactive component, respectively. Morphology and structure of bioactive catechin-gelatin nanofiber were characterized by scanning electron microscopy (SEM) and fourier transform infrared spectroscopy (FTIR), respectively. SEM analysis showed that morphology of nanofiber composite was smooth and had average diameter 398.97 nm. FTIR analysis results were used to confirm structure of catechin-gelatin nanofiber. It was confirmed by FTIR that specific vibration band peak amide A (N-H) at 3286,209 cm-1, amide B (N-H) 3069,396 cm-1, amide I (C=O) at 1643,813 cm-1, amide II (N-H and CN) at 1538,949 cm-1, amide III (C-N) at 1276,789 cm-1, C-O-C from polyethylene oxide at 1146,418 cm-1, respectively. When examined to S. Aureus bacteria, Ag/catechin/gelatin nanofiber show inhabitation performance around 40.44%. Ag(NP)/catechin/gelatin nanofiber has potential application antibacterial medical application.

OPTIMIZATION OF MARSHMALLOW GELATIN FUNCTIONAL PURPOSE

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G. O. Magomedov

2015-01-01

Full Text Available Marshmallow on gelatin or marshmallow items finely porous foam structure. This product is a popular and widely demanded confectionery products to the public, especially children and students. Urgent by the NE-a method for producing a mini-marshmallow functionality in terms of domestic production with natural-functional ingredient – gum-arabic «Instantgum», which is the source of natural-fiber, improving gastrointestinal function, lowering blood glucose levels that especially important in the use of products of diabetics. The inclusion of gum-arabic in the diet leads to a prebiotic effect, stimulating human immune system and improves the functioning intestines. Technology mini marshmallow gelatin with gum-arabic comprises the following manufacturing steps of: preparing a gelatin solution, an aqueous solution of gum-arabic, sugar syrup, invert syrup, whipped gelatinous mass density of 400-600 kg / m3, molding or pressing off mass fractionation followed by cooling and cutting, exposure and dusting starch and sugar mixture housings mini marshmallows. The optimization formulation mini marshmallows on gelatin with functional ingredients «Instantgum» based on gum-arabic method of multicriteria optimization of op-using desirability function Harrington. The greatest value generalized desirability function had a sample with 10% in-functional ingredient, since it belongs to the best set of parameters optimization. The developed product has high organoleptic, physical and chemical parameters, satisfactory form-retaining ability elastic chewing consistency and a functional purpose by NIJ-containing dietary fiber. Proposed can be recommended for the production on-time with continuous mechanized lines to develop Pastila products.

Development and characterization of coaxially electrospun gelatin coated poly (3-hydroxybutyric acid) thin films as potential scaffolds for skin regeneration

International Nuclear Information System (INIS)

Nagiah, Naveen; Madhavi, Lakshmi; Anitha, R.; Anandan, C.; Srinivasan, Natarajan Tirupattur; Sivagnanam, Uma Tirichurapalli

2013-01-01

The morphology of fibers synthesized through electrospinning has been found to mimic extracellular matrix. Coaxially electrospun fibers of gelatin (sheath) coated poly (3-hydroxybutyric acid) (PHB) (core) was developed using 2,2,2 trifluoroethanol(TFE) and 1,1,1,3,3,3 hexafluoro-2-propanol(HFIP) as solvents respectively. The coaxial structure and coating of gelatin with PHB fibers was confirmed through transmission electron microscopy (TEM), scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). Thermal stability of the coaxially electrospun fibers was analyzed using thermogravimetric analysis(TGA), differential scanning calorimetry(DSC) and differential thermogravimetric analysis(DTA). Complete evaporation of solvent and gelatin grafting over PHB fibers was confirmed through attenuated total reflection-Fourier transformed infrared spectroscopy (ATR-FTIR). The coaxially electrospun fibers exhibited competent tensile properties for skin regeneration with high surface area and porosity. In vitro degradation studies proved the stability of fibers and its potential applications in tissue engineering. The fibers supported the growth of human dermal fibroblasts and keratinocytes with normal morphology indicating its potential as a scaffold for skin regeneration. - Highlights: • Coaxial electrospinning was employed to develop core-shell fibers of PHB and gelatin. • The scaffold has competent physicochemical properties. • Developed scaffold will have high impact as a dermal substitute in skin regeneration

Development and characterization of coaxially electrospun gelatin coated poly (3-hydroxybutyric acid) thin films as potential scaffolds for skin regeneration

Energy Technology Data Exchange (ETDEWEB)

Nagiah, Naveen [Bioproducts Lab, Central Leather Research Institute, Adyar, Chennai (India); Madhavi, Lakshmi; Anitha, R. [CavinKare Research Centre, Ekkattuthangal, Chennai (India); Anandan, C. [National Aerospace Laboratory, Banglore (India); Srinivasan, Natarajan Tirupattur, E-mail: naveen.nagiah@gmail.com [Conducting Polymers Lab, Department of Physics, Indian Institute of Technology Madras, Chennai (India); Sivagnanam, Uma Tirichurapalli, E-mail: suma67@gmail.com [Bioproducts Lab, Central Leather Research Institute, Adyar, Chennai (India)

2013-10-01

The morphology of fibers synthesized through electrospinning has been found to mimic extracellular matrix. Coaxially electrospun fibers of gelatin (sheath) coated poly (3-hydroxybutyric acid) (PHB) (core) was developed using 2,2,2 trifluoroethanol(TFE) and 1,1,1,3,3,3 hexafluoro-2-propanol(HFIP) as solvents respectively. The coaxial structure and coating of gelatin with PHB fibers was confirmed through transmission electron microscopy (TEM), scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). Thermal stability of the coaxially electrospun fibers was analyzed using thermogravimetric analysis(TGA), differential scanning calorimetry(DSC) and differential thermogravimetric analysis(DTA). Complete evaporation of solvent and gelatin grafting over PHB fibers was confirmed through attenuated total reflection-Fourier transformed infrared spectroscopy (ATR-FTIR). The coaxially electrospun fibers exhibited competent tensile properties for skin regeneration with high surface area and porosity. In vitro degradation studies proved the stability of fibers and its potential applications in tissue engineering. The fibers supported the growth of human dermal fibroblasts and keratinocytes with normal morphology indicating its potential as a scaffold for skin regeneration. - Highlights: • Coaxial electrospinning was employed to develop core-shell fibers of PHB and gelatin. • The scaffold has competent physicochemical properties. • Developed scaffold will have high impact as a dermal substitute in skin regeneration.

RENDEMEN DAN KOMPOSISI PROKSIMAT GELATIN KULIT IKAN BELUT DAN LELE PADA KEADAAN SEGAR DAN KERING (YIELD AND PROXIMATE OF GELATIN EXTRACTED FROM FRESH AND DRY SWAMP ELL AND CATFISH SKIN

Directory of Open Access Journals (Sweden)

Hafni Rahmawati

2017-02-01

Full Text Available Kulit ikan belut dan lele berpotensi untuk diekstrak gelatinnya. Kulit ikan belut dan lele tidak bersisik, berlendir dan berlemak untuk ikan lele, berbeda dengan kulit ikan pada umumnya yang dijadikan gelatin. Penelitian ini mempelajari tentang gelatin kulit ikan belut dan lele dari segi rendemen dan komposisi proksimat. Untuk mengetahui pengaruh pengeringan, kondisi kulit segar dan kering juga dipelajari. Tahapan ekstraksi yang dilakukan untuk keseluruhan jenis ikan sama, kecuali penanganan kulit ikan kering yang sebelumnya direndam dalam air selama 4 jam. Kulit ikan direndam kembali dalam 0,05M asam asetat selama 10 jam, kemudian dicuci dan diekstraksi dengan aquadest pada suhu 80OC selama 2 jam, cairan yang didapat difiltrasi. Filtratnya dikeringkan dalam cabinet dryer suhu 55OC selama        48 jam hingga diperoleh lembaran gelatin, kemudian diblender menjadi granula gelatin. Hasil penelitian menunjukkan bahwa gelatin kulit ikan kering mempunyai rendemen sedikit lebih rendah dibandingkan gelatin dari kulit ikan segarnya. Kadar protein gelatin kulit ikan kering telihat tinggi dibandingkan kulit segarnya. Pengeringan kulit ikan berpengaruh pada penurunan kadar abu dan lemak, namun tidak mempengaruhi kadar air gelatin hasil ekstraksi. Gelatin kulit segar ikan lele memiliki nilai rendemen tertinggi yaitu 22,01%. Komposisi proksimat yang terbaik diantara keseluruhan kondisi dan jenis kulit dapat dilihat pada gelatin kulit segar ikan belut dimana memiliki kadar air 9,91%; kadar abu 3,07%; kadar protein 91,61%; dan kadar lemak 0,82%. Skin of swam ell and catfish were potential to gelatin extracted. Swamp ell skin doesn’t have scales, with much mucus and a few fat for catfish, it’s different from another fish skin that gelatin extracted usually. This research was studied yield and proximate gelatin extracted from skin of swam ell and catfish. Influence of drying was observed too. The first stage of the research was raw

Gelatin/hydroxypropyl methylcellulose matrices — Polymer interactions approach for oral disintegrating films

Energy Technology Data Exchange (ETDEWEB)

Tedesco, Marcela P., E-mail: marcela.tedesco@usp.br; Monaco-Lourenço, Carla A., E-mail: carla.monaco@usp.br; Carvalho, Rosemary A., E-mail: rosecarvalho@usp.br

2016-12-01

Oral disintegrating film represents an optimal alternative for delivery system of active compounds. The choice of film-forming polymer is the first step in the development of oral disintegrating films and the knowledge of molecular interactions in this matrix is fundamental to advance in this area. Therefore, this study aimed to characterize gelatin and hydroxypropyl methylcellulose (HPMC) films and their blends as matrices of oral disintegrating films. The films were produced by casting technique and were characterized by scanning electron microscopy, Fourier transform infrared spectroscopy, differential scanning calorimetry, mechanical properties, contact angle, time disintegration and bioadhesive strength. Differential scanning calorimetry showed that enthalpy of fusion and melting temperatures of the blends films were lower than those of the gelatin film, which may be associated with the lack of intra-chain interactions also observed in the Fourier transform infrared spectra. In blends, a less compact cross-section structure was observed in scanning electron microscopy images compared with isolated polymer films. The addition of HPMC increased the elongation, hydrophilicity and in vitro bioadhesive force and decreased in vitro disintegration time, important properties in the development of oral disintegrating films. Although the mixture of the polymers showed no synergistic behavior, this study may contribute to the development of new applications for polymeric matrices in the pharmaceutical industry. - Highlights: • Effect of gelatin/HPMC blend on oral disintegrating films properties. • Intramolecular interaction of films was evaluated. • Mixtures of the polymers provoked reductions in intra-chain interactions. • HPMC content affects hydrophilic of oral disintegrating films.

Gelatin/hydroxypropyl methylcellulose matrices — Polymer interactions approach for oral disintegrating films

International Nuclear Information System (INIS)

Tedesco, Marcela P.; Monaco-Lourenço, Carla A.; Carvalho, Rosemary A.

2016-01-01

Oral disintegrating film represents an optimal alternative for delivery system of active compounds. The choice of film-forming polymer is the first step in the development of oral disintegrating films and the knowledge of molecular interactions in this matrix is fundamental to advance in this area. Therefore, this study aimed to characterize gelatin and hydroxypropyl methylcellulose (HPMC) films and their blends as matrices of oral disintegrating films. The films were produced by casting technique and were characterized by scanning electron microscopy, Fourier transform infrared spectroscopy, differential scanning calorimetry, mechanical properties, contact angle, time disintegration and bioadhesive strength. Differential scanning calorimetry showed that enthalpy of fusion and melting temperatures of the blends films were lower than those of the gelatin film, which may be associated with the lack of intra-chain interactions also observed in the Fourier transform infrared spectra. In blends, a less compact cross-section structure was observed in scanning electron microscopy images compared with isolated polymer films. The addition of HPMC increased the elongation, hydrophilicity and in vitro bioadhesive force and decreased in vitro disintegration time, important properties in the development of oral disintegrating films. Although the mixture of the polymers showed no synergistic behavior, this study may contribute to the development of new applications for polymeric matrices in the pharmaceutical industry. - Highlights: • Effect of gelatin/HPMC blend on oral disintegrating films properties. • Intramolecular interaction of films was evaluated. • Mixtures of the polymers provoked reductions in intra-chain interactions. • HPMC content affects hydrophilic of oral disintegrating films.

Histone deacetylase 3 supports endochondral bone formation by controlling cytokine signaling and matrix remodeling

Science.gov (United States)

Carpio, Lomeli R.; Bradley, Elizabeth W.; McGee-Lawrence, Meghan E.; Weivoda, Megan M.; Poston, Daniel D.; Dudakovic, Amel; Xu, Ming; Tchkonia, Tamar; Kirkland, James L.; van Wijnen, Andre J.; Oursler, Merry Jo; Westendorf, Jennifer J.

2017-01-01

Histone deacetylase (HDAC) inhibitors are efficacious epigenetic-based therapies for some cancers and neurological disorders; however, each of these drugs inhibits multiple HDACs and has detrimental effects on the skeleton. To better understand how HDAC inhibitors affect endochondral bone formation, we conditionally deleted one of their targets, Hdac3, pre- and postnatally in type II collagen α1 (Col2α1)–expressing chondrocytes. Embryonic deletion was lethal, but postnatal deletion of Hdac3 delayed secondary ossification center formation, altered maturation of growth plate chondrocytes, and increased osteoclast activity in the primary spongiosa. HDAC3-deficient chondrocytes exhibited increased expression of cytokine and matrix-degrading genes (Il-6, Mmp3, Mmp13, and Saa3) and a reduced abundance of genes related to extracellular matrix production, bone development, and ossification (Acan, Col2a1, Ihh, and Col10a1). Histone acetylation increased at and near genes that had increased expression. The acetylation and activation of nuclear factor κB (NF-κB) were also increased in HDAC3-deficient chondrocytes. Increased cytokine signaling promoted autocrine activation of Janus kinase (JAK)–signal transducer and activator of transcription (STAT) and NF-κB pathways to suppress chondrocyte maturation, as well as paracrine activation of osteoclasts and bone resorption. Blockade of interleukin-6 (IL-6)–JAK–STAT signaling, NF-κB signaling, and bromodomain extraterminal proteins, which recognize acetylated lysines and promote transcriptional elongation, significantly reduced Il-6 and Mmp13 expression in HDAC3-deficient chondrocytes and secondary activation in osteoclasts. The JAK inhibitor ruxolitinib also reduced osteoclast activity in Hdac3 conditional knockout mice. Thus, HDAC3 controls the temporal and spatial expression of tissue-remodeling genes and inflammatory responses in chondrocytes to ensure proper endochondral ossification during development. PMID

Hydroxyapatite fiber reinforced poly(alpha-hydroxy ester) foams for bone regeneration

Science.gov (United States)

Thomson, R. C.; Yaszemski, M. J.; Powers, J. M.; Mikos, A. G.; McIntire, L. V. (Principal Investigator)

1998-01-01

A process has been developed to manufacture biodegradable composite foams of poly(DL-lactic-co-glycolic acid) (PLGA) and hydroxyapatite short fibers for use in bone regeneration. The processing technique allows the manufacture of three-dimensional foam scaffolds and involves the formation of a composite material consisting of a porogen material (either gelatin microspheres or salt particles) and hydroxyapatite short fibers embedded in a PLGA matrix. After the porogen is leached out, an open-cell composite foam remains which has a pore size and morphology defined by the porogen. By changing the weight fraction of the leachable component it was possible to produce composite foams with controlled porosities ranging from 0.47 +/- 0.02 to 0.85 +/- 0.01 (n = 3). Up to a polymer:fiber ratio of 7:6, short hydroxyapatite fibers served to reinforce low-porosity PLGA foams manufactured using gelatin microspheres as a porogen. Foams with a compressive yield strength up to 2.82 +/- 0.63 MPa (n = 3) and a porosity of 0.47 +/- 0.02 (n = 3) were manufactured using a polymer:fiber weight ratio of 7:6. In contrast, high-porosity composite foams (up to 0.81 +/- 0.02, n = 3) suitable for cell seeding were not reinforced by the introduction of increasing quantities of hydroxyapatite short fibers. We were therefore able to manufacture high-porosity foams which may be seeded with cells but which have minimal compressive yield strength, or low porosity foams with enhanced osteoconductivity and compressive yield strength.

Osteoclasts prefer aged bone

DEFF Research Database (Denmark)

Henriksen, K; Leeming, Diana Julie; Byrjalsen, I

2007-01-01

We investigated whether the age of the bones endogenously exerts control over the bone resorption ability of the osteoclasts, and found that osteoclasts preferentially develop and resorb bone on aged bone. These findings indicate that the bone matrix itself plays a role in targeted remodeling...... of aged bones....

Structure evolution of gelatin particles induced by pH and ionic strength.

Science.gov (United States)

Xu, Jing; Li, Tianduo; Tao, Furong; Cui, Yuezhi; Xia, Yongmei

2013-03-01

Microstructure of gelatin particles played a key role in determining the physicochemical properties of gelatin. Ionic strength and pH as systematic manners were considered to affect gelatin particles structure on the micrometer scale. Scanning electron microscopy was used for depicting the morphologies of gelatin particles. Increasing pH to 10.0 or decreasing pH to 4.0, spherical, spindle, and irregular aggregates of gelatin particles at 2, 6, 10, and 14% solution (w/w) were all transformed to spindle aggregates. When NaCl was added to the system, the molecular chains of gelatin possibly rearranged themselves in a stretched state, and the ribbon aggregates was observed. The structural transitions of gelatin aggregates were strongly depended on the electrostatic repulsion. In the gelatin-sodium dodecyl sulfate (SDS) case, the micrometer scale of aggregates was larger and the different degrees of cross-links were induced through hydrophobic interaction and electrostatic repulsion. Copyright © 2012 Wiley Periodicals, Inc.

Matrix metalloproteinase-12 (MMP-12) in osteoclasts

DEFF Research Database (Denmark)

Hou, Peng; Troen, Tine; Ovejero, Maria C

2004-01-01

Osteoclasts require matrix metalloproteinase (MMP) activity and cathepsin K to resorb bone, but the critical MMP has not been identified. Osteoclasts express MMP-9 and MMP-14, which do not appear limiting for resorption, and the expression of additional MMPs is not clear. MMP-12, also called...... bone show MMP-12 expression in osteoclasts in calvariae and long bones. We also demonstrate that recombinant MMP-12 cleaves the putative functional domains of osteopontin and bone sialoprotein, two bone matrix proteins that strongly influence osteoclast activities, such as attachment, spreading...

Gelatin Nanoparticles with Enhanced Affinity for Calcium Phosphate

NARCIS (Netherlands)

Farbod, K.; Diba, M.; Zinkevich, T.; Schmidt, S.; Harrington, M.J.; Kentgens, A.P.; Leeuwenburgh, S.C.

2016-01-01

Gelatin nanoparticles can be tuned with respect to their drug loading efficiency, degradation rate, and release kinetics, which renders these drug carriers highly suitable for a wide variety of biomedical applications. The ease of functionalization has rendered gelatin an interesting candidate

Preparation and Characterization of Gelatin Nanofibers Containing Silver Nanoparticles

Science.gov (United States)

Jeong, Lim; Park, Won Ho

2014-01-01

Ag nanoparticles (NPs) were synthesized in formic acid aqueous solutions through chemical reduction. Formic acid was used for a reducing agent of Ag precursor and solvent of gelatin. Silver acetate, silver tetrafluoroborate, silver nitrate, and silver phosphate were used as Ag precursors. Ag+ ions were reduced into Ag NPs by formic acid. The formation of Ag NPs was characterized by a UV-Vis spectrophotometer. Ag NPs were quickly generated within a few minutes in silver nitrate (AgNO3)/formic acid solution. As the water content of formic acid aqueous solution increased, more Ag NPs were generated, at a higher rate and with greater size. When gelatin was added to the AgNO3/formic acid solution, the Ag NPs were stabilized, resulting in smaller particles. Moreover, gelatin limits further aggregation of Ag NPs, which were effectively dispersed in solution. The amount of Ag NPs formed increased with increasing concentration of AgNO3 and aging time. Gelatin nanofibers containing Ag NPs were fabricated by electrospinning. The average diameters of gelatin nanofibers were 166.52 ± 32.72 nm, but these decreased with the addition of AgNO3. The average diameters of the Ag NPs in gelatin nanofibers ranged between 13 and 25 nm, which was confirmed by transmission electron microscopy (TEM). PMID:24758929

Effects of Long-term exposure of Gelatinated and Non-gelatinated Cadmium Telluride Quantum Dots on Differentiated PC12 cells

LENUS (Irish Health Repository)

Prasad, Babu R

2012-01-20

Abstract Background The inherent toxicity of unmodified Quantum Dots (QDs) is a major hindrance to their use in biological applications. To make them more potent as neuroprosthetic and neurotherapeutic agents, thioglycolic acid (TGA) capped CdTe QDs, were coated with a gelatine layer and investigated in this study with differentiated pheochromocytoma 12 (PC12) cells. The QD - cell interactions were investigated after incubation periods of up to 17 days by MTT and APOTOX-Glo Triplex assays along with using confocal microscopy. Results Long term exposure (up to 17 days) to gelatinated TGA-capped CdTe QDs of PC12 cells in the course of differentiation and after neurites were grown resulted in dramatically reduced cytotoxicity compared to non-gelatinated TGA-capped CdTe QDs. Conclusion The toxicity mechanism of QDs was identified as caspase-mediated apoptosis as a result of cadmium leaking from the core of QDs. It was therefore concluded that the gelatine capping on the surface of QDs acts as a barrier towards the leaking of toxic ions from the core QDs in the long term (up to 17 days).

Oxygen Plasma Treatment on 3D-Printed Chitosan/Gelatin/Hydroxyapatite Scaffolds for Bone Tissue Engineering.

Science.gov (United States)

Lee, Chang-Min; Yang, Seong-Won; Jung, Sang-Chul; Kim, Byung-Hoon

2017-04-01

The 3D hydroxyapatite/gelatin/chitosan composite scaffolds were fabricated by 3D printing technique. The scaffolds were treated by oxygen plasma to improve the bioactivity and its surface characterization and in vitro cell culture were investigated. The scaffolds exhibited the good porosity and interconnectivity of pores. After oxygen plasma etching, roughness and wettability on the scaffolds surface are increased. Plasma treated scaffolds showed higher proliferation than that of untreated scaffolds. Oxygen plasma treatment could be used as potential tool to enhance the biocompatibility on the 3D composite scaffolds.

Evaluation of tilapia skin gelatin as a mammalian gelatin replacer in acid milk gels and low-fat stirred yogurt.

Science.gov (United States)

Pang, Zhihua; Deeth, Hilton; Yang, Hongshun; Prakash, Sangeeta; Bansal, Nidhi

2017-05-01

Tilapia skin gelatin (TSG) was studied in a 3-stage process (cooling, annealing, and heating) for pure gelatin gels and in a 4-stage process (acidification, cooling, annealing, and heating) for acid milk gels and cultured yogurt. The aim was to evaluate the use of TSG as a replacement for mammalian gelatin in yogurt. In pure TSG gels, stronger gels with higher melting temperatures were formed with increasing TSG concentrations. Compared with bovine gelatin (BG), which gelled at a concentration of 2.5%, TSG gels had lower gelling (14.1°C) and melting (24°C) temperatures but comparable storage moduli during annealing. In acid milk gels, addition of TSG increased the firmness of the gels with increasing concentration. Gelling and melting points of TSG in milk gels were observed at sufficient concentrations during cooling and heating. Strands and sheets were observed in the electron micrographs of milk gels with 1% TSG and a very dense structure was observed with 2.5% TSG. Yogurt with 0.4% TSG had similar viscosity, consistency, pseudoplasticity, and thixotropy as yogurt containing 0.4% BG; no difference was perceived by sensory panelists according to a triangle test. Addition of 0.4% TSG completely prevented whey separation from the acid milk gel and yogurt. The results suggest that TSG could be a suitable replacement for mammalian gelatin in low-fat stirred yogurt. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Comparison of water gel desserts from fish skin and pork gelatins using instrumental measurements.

Science.gov (United States)

Zhou, Peng; Regenstein, Joe M

2007-05-01

The objective of this study was to compare water gel desserts from various gelatins using instrumental measurements. The puncture test and texture profile analysis (TPA) with compression were determined at 25% and 75% deformation; the melting properties were determined rheologically by monitoring the change of storage modulus (G') with increasing temperature. The measurements with 25% deformation were always nondestructive, while measurements with 75% deformation were mostly destructive. Desserts made from Alaska pollock gelatin (AG) or gelatin mixtures containing AG were more resistant to the destruction caused by the large deformation than tilapia gelatin and pork gelatins. In addition, the gel dessert made from AG melted at a lower temperature than those from tilapia skin gelatin and pork gelatins, while desserts made from gelatin mixtures reflected the melting properties of the separate gelatins.

Evaluation of cross-linked gelatin membranes as delivery carriers for retinal sheets

Energy Technology Data Exchange (ETDEWEB)

Lai, Jui-Yang, E-mail: jylai@mail.cgu.edu.tw [Institute of Biochemical and Biomedical Engineering, Chang Gung University, Taoyuan, 33302 Taiwan (China); Biomedical Engineering Research Center, Chang Gung University, Taoyuan, 33302 Taiwan (China); Molecular Medicine Research Center, Chang Gung University, Taoyuan, 33302 Taiwan (China); Li, Ya-Ting [Institute of Biochemical and Biomedical Engineering, Chang Gung University, Taoyuan, 33302 Taiwan (China)

2010-06-15

The delivery of intact sheet transplants to the subretinal space can prevent cell loss that is generally associated with the injection of cell suspensions or cell aggregates. The aim of this study was to develop chemically modified gelatin matrices that enhance the delivery efficiency and analyze whether the gelatin membranes cross-linked with 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) can be considered as potential carriers for retinal sheets. The characteristics of EDC cross-linked gelatin membranes were determined by mechanical and in vitro degradation tests, melting point measurements, cell proliferation assays, cytokine expression analyses, and tissue delivery studies. Gelatin membranes without cross-linking and glutaraldehyde cross-linked gelatin samples were used for comparison. Results of this study indicated that introduction of cross-links is capable of rendering the gelatin network more stable against mechanical stresses and deformations as well as rapid hydrolysis during intraocular delivery of delicate tissue sheets. In comparison with the glutaraldehyde treated samples, the EDC cross-linked gelatin membranes showed a better degradation profile and a relatively higher cytocompatibility. In addition, after EDC cross-linking, the gelatin matrices having an acceptable melting point could be used for the fabrication of a sandwich-like carrier with a high transfer and encapsulation efficiency. These findings suggest that the cross-linking agent type gives an influence on delivery functionality of gelatin membranes. In summary, the EDC cross-linked gelatin is an ideal candidate for use as a carrier material in retinal sheet delivery applications.

Wet method for measuring starch gelatinization temperature using electrical conductivity.

Science.gov (United States)

Morales-Sanchez, E; Figueroa, J D C; Gaytan-Martínez, M

2009-09-01

The objective of the present study was to develop a method for obtaining the gelatinization temperature of starches by using electrical conductivity. Native starches from corn, rice, potato, and wheat were prepared with different proportions of water and heated from room temperature to 90 degrees C, in a device especially designed for monitoring the electrical conductivity as a function of temperature. The results showed a linear trend of the electrical conductivity with the temperature until it reaches the onset gelatinization temperature. After that point, the electrical conductivity presented an increment or decrement depending on the water content in the sample and it was related to starch swelling and gelatinization phenomena. At the end gelatinization temperature, the conductivity becomes stable and linear, indicating that there are no more changes of phase. The starch gelatinization parameter, which was evaluated in the 4 types of starches using the electrical conductivity, was compared with those obtained by using differential scanning calorimeter (DSC). The onset temperature at which the electrical conductivity increased or decreased was found to be similar to that obtained by DSC. Also, the final temperature at which the electrical conductivity returned to linearity matched the end gelatinization temperature of the DSC. Further, a wet method for measuring the onset, peak, and end gelatinization temperatures as a function of temperature using the electrical conductivity curves is presented for a starch-water suspension.

Cell expansion of human articular chondrocytes on macroporous gelatine scaffolds-impact of microcarrier selection on cell proliferation

Energy Technology Data Exchange (ETDEWEB)

Pettersson, Sofia; Kratz, Gunnar [Laboratory for Reconstructive Plastic Surgery, Department of Clinical and Experimental Medicine, Linkoeping University, SE-581 85 Linkoeping (Sweden); Wetteroe, Jonas [Rheumatology/AIR, Department of Clinical and Experimental Medicine, Linkoeping University, SE-581 85 Linkoeping (Sweden); Tengvall, Pentti, E-mail: sofia.pettersson@liu.se [Institute of Clinical Sciences, Department of Biomaterials, The Sahlgrenska Academy at University of Gothenburg, SE-405 30 Gothenburg (Sweden)

2011-12-15

This study investigates human chondrocyte expansion on four macroporous gelatine microcarriers (CultiSpher) differing with respect to two manufacturing processes-the amount of emulsifier used during initial preparation and the gelatine cross-linking medium. Monolayer-expanded articular chondrocytes from three donors were seeded onto the microcarriers and cultured in spinner flask systems for a total of 15 days. Samples were extracted every other day to monitor cell viability and establish cell counts, which were analysed using analysis of variance and piecewise linear regression. Chondrocyte densities increased according to a linear pattern for all microcarriers, indicating an ongoing, though limited, cell proliferation. A strong chondrocyte donor effect was seen during the initial expansion phase. The final cell yield differed significantly between the microcarriers and our results indicate that manufacturing differences affected chondrocyte densities at this point. Remaining cells stained positive for chondrogenic markers SOX-9 and S-100 but extracellular matrix formation was modest to undetectable. In conclusion, the four gelatine microcarriers supported chondrocyte adhesion and proliferation over a two week period. The best yield was observed for microcarriers produced with low emulsifier content and cross-linked in water and acetone. These results add to the identification of optimal biomaterial parameters for specific cellular processes and populations.

Soft gelatin capsules (softgels).

Science.gov (United States)

Gullapalli, Rampurna Prasad

2010-10-01

It is estimated that more than 40% of new chemical entities (NCEs) coming out of the current drug discovery process have poor biopharmaceutical properties, such as low aqueous solubility and/or permeability. These suboptimal properties pose significant challenges for the oral absorption of the compounds and for the development of orally bioavailable dosage forms. Development of soft gelatin capsule (softgel) dosage form is of growing interest for the oral delivery of poorly water soluble compounds (BCS class II or class IV). The softgel dosage form offers several advantages over other oral dosage forms, such as delivering a liquid matrix designed to solubilize and improve the oral bioavailability of a poorly soluble compound as a unit dose solid dosage form, delivering low and ultra-low doses of a compound, delivering a low melting compound, and minimizing potential generation of dust during manufacturing and thereby improving the safety of production personnel. However, due to the very dynamic nature of the softgel dosage form, its development and stability during its shelf-life are fraught with several challenges. The goal of the current review is to provide an in-depth discussion on the softgel dosage form to formulation scientists who are considering developing softgels for therapeutic compounds.

Dehydration of pollock skin prior to gelatin production

Science.gov (United States)

Alaska pollock (Theragra chalcogramma) is the U.S.A.'s largest commercial fishery, with an annual catch of over 1 million tons. During pollock processing, the skins are discarded or made into fish meal, despite their value for gelatin production. The absence of gelatin-processing facilities in Alask...

Extraction and characterisation of gelatin from the skin of striped ...

African Journals Online (AJOL)

Functional properties of gelatin from skin of striped catfish with and without bleaching for 48 h by 5% H2O2 (w/v)) were studied. Gelatin from skin bleached with 5% H2O2 for 48 h showed the highest yield (16.18 g). Bleaching not only improved the colour of gelatin gel by increasing the L* (lightness)-value and decreasing ...

Biomimetic Synthesis of Gelatin Polypeptide-Assisted Noble-Metal Nanoparticles and Their Interaction Study

Science.gov (United States)

Liu, Ying; Liu, Xiaoheng; Wang, Xin

2011-12-01

Herein, the generation of gold, silver, and silver-gold (Ag-Au) bimetallic nanoparticles was carried out in collagen (gelatin) solution. It first showed that the major ingredient in gelatin polypeptide, glutamic acid, acted as reducing agent to biomimetically synthesize noble metal nanoparticles at 80°C. The size of nanoparticles can be controlled not only by the mass ratio of gelatin to gold ion but also by pH of gelatin solution. Interaction between noble-metal nanoparticles and polypeptide has been investigated by TEM, UV-visible, fluorescence spectroscopy, and HNMR. This study testified that the degradation of gelatin protein could not alter the morphology of nanoparticles, but it made nanoparticles aggregated clusters array (opposing three-dimensional α-helix folding structure) into isolated nanoparticles stabilized by gelatin residues. This is a promising merit of gelatin to apply in the synthesis of nanoparticles. Therefore, gelatin protein is an excellent template for biomimetic synthesis of noble metal/bimetallic nanoparticle growth to form nanometer-sized device.

Human alveolar bone cell proliferation, expression of osteoblastic phenotype, and matrix mineralization on porous titanium produced by powder metallurgy.

Science.gov (United States)

Rosa, Adalberto Luiz; Crippa, Grasiele Edilaine; de Oliveira, Paulo Tambasco; Taba, Mario; Lefebvre, Louis-Philippe; Beloti, Marcio Mateus

2009-05-01

This study aimed at investigating the influence of the porous titanium (Ti) structure on the osteogenic cell behaviour. Porous Ti discs were fabricated by the powder metallurgy process with the pore size typically between 50 and 400 microm and a porosity of 60%. Osteogenic cells obtained from human alveolar bone were cultured until subconfluence and subcultured on dense Ti (control) and porous Ti for periods of up to 17 days. Cultures grown on porous Ti exhibited increased cell proliferation and total protein content, and lower levels of alkaline phosphatase (ALP) activity than on dense Ti. In general, gene expression of osteoblastic markers-runt-related transcription factor 2, collagen type I, alkaline phosphatase, bone morphogenetic protein-7, and osteocalcin was lower at day 7 and higher at day 17 in cultures grown on porous Ti compared with dense Ti, a finding consistent with the enhanced growth rate for such cultures. The amount of mineralized matrix was greater on porous Ti compared with the dense one. These results indicate that the porous Ti is an appropriate substrate for osteogenic cell adhesion, proliferation, and production of a mineralized matrix. Because of the three-dimensional environment it provides, porous Ti should be considered an advantageous substrate for promoting desirable implant surface-bone interactions.

Premature loss of bone remodeling compartment canopies is associated with deficient bone formation

DEFF Research Database (Denmark)

Jensen, Pia Rosgaard; Andersen, Thomas Levin; Søe, Kent

2011-01-01

A remarkable property of bone remodeling is that osteoblasts form bone matrix exactly where and when osteoclasts have removed it. The bone remodeling compartment (BRC) canopies that cover bone surfaces undergoing remodeling, were proposed to be critical players in this mechanism. Here, we provide...

The characterisation of a genipin-gelatin gel dosimeter

International Nuclear Information System (INIS)

Davies, J B; Bosi, S; Baldock, C

2010-01-01

Genipin cross links gelatin to slowly form a blue colour that bleaches upon irradiation. Spectrophotometric measurements of the absorbance change following irradiation to doses up to 100 Gy gives a linear dose response for certain concentrations of the gel ingredients; genipin, gelatin and sulphuric acid. Dose sensitivity increases with increasing concentrations of sulphuric acid and genipin and is also strongly dependent on the time allowed for the genipin-gelatin cross linking reaction (referred to here as blending) to take place. The optimum formulation of this gel was found for genipin concentration between 0.3 - 0.5 mM and blending time of at least 4 h.

Gelatin/piassava composites treated by Electron Beam Radiation

International Nuclear Information System (INIS)

Takinami, Patricia Yoko Inamura; Shimazaki, Kleber; Moura, Esperidiana Augusta Barretos de; Mastro, Nelida Lucia del; Colombo, Maria Aparecida

2010-01-01

Piassava (Attalea funifera Mart) fiber has been investigated as reinforcement for polymer composites with potential for practical applications. The purpose of the present work was to assess the behavior of specimens of piassava fiber and gelatin irradiated with electron beam at different doses and percentage. The piassava/gelatin specimens were made with 5 and 10% (w/w) piassava fiber, gelatin 25% (w/w), glycerin as plasticizer and acrylamide as copolymer. The samples were irradiated up to 40 kGy using an electron beam accelerator, at room temperature in presence of air. Preliminary results showed mechanical properties enhancement with the increase in radiation dose. (author)

Heterotopic bone formation in the musculus latissimus dorsi of sheep using β-tricalcium phosphate scaffolds: evaluation of an extended prefabrication time on bone formation and matrix degeneration.

Science.gov (United States)

Spalthoff, S; Jehn, P; Zimmerer, R; Möllmann, U; Gellrich, N-C; Kokemueller, H

2015-06-01

We previously generated viable heterotopic bone in living animals and found that 3 months of intrinsic vascularization improved bone formation and matrix degeneration. In this study, we varied the pre-vascularization time to determine its effects on the kinetics of bone formation and ceramic degradation. Two 25-mm-long cylindrical β-tricalcium phosphate scaffolds were filled intraoperatively with autogenous iliac crest bone marrow and implanted in the latissimus dorsi muscle in six sheep. To examine the effect of axial perfusion, one scaffold was surgically implanted with (group C) or without (group D) a central vascular bundle. All animals were sacrificed 6 months postoperatively and histomorphometric measurements were compared to previous results. All implanted scaffolds exhibited ectopic bone growth. However, bone growth was not significantly different between the 3-month (group A, 0.191±0.097 vs. group C, 0.237±0.075; P=0.345) and 6-month (group B, 0.303±0.105 vs. group D, 0.365±0.258; P=0.549) pre-vascularization durations, regardless of vessel supply; early differences between surgically and extrinsically vascularized constructs disappeared after 6 months. Here, we describe a reliable procedure for generating ectopic bone in vivo. A 3-month pre-vascularization duration appears sufficient and ceramic degradation proceeds in accordance with bone generation, supporting the hypothesis of cell-mediated resorption. Copyright © 2014 International Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved.

A combined approach of enamel matrix derivative gel and autogenous bone grafts in treatment of intrabony periodontal defects. A case report.

Science.gov (United States)

Leung, George; Jin, Lijian

2003-04-01

Enamel matrix derivative (EMD) has recently been introduced as a new modality in regenerative periodontal therapy. This case report demonstrates a combined approach in topical application of EMD gel (Emdogain) and autogenous bone grafts for treatment of intrabony defects and furcation involvement defects in a patient with chronic periodontitis. The seven-month post-surgery clinical and radiographic results were presented. The combined application of EMD gel with autogenous bone grafts in intrabony osseous defects resulted in clinically significant gain of attachment on diseased root surfaces and bone fill on radiographs. Further controlled clinical studies are required to confirm the long-term effectiveness of the combination of EMD gel and autogenous bone grafts in treatment of various osseous defects in subjects with chronic periodontitis.

Fabrication of Aligned Carbon Nanotube/Polycaprolactone/Gelatin Nanofibrous Matrices for Schwann Cell Immobilization

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Shiao-Wen Tsai

2014-01-01

Full Text Available In this study, we utilized a mandrel rotating collector consisting of two parallel, electrically conductive pieces of tape to fabricate aligned electrospun polycaprolactone/gelatin (PG and carbon nanotube/polycaprolactone/gelatin (PGC nanofibrous matrices. Furthermore, we examined the biological performance of the PGC nanofibrous and film matrices using an in vitro culture of RT4-D6P2T rat Schwann cells. Using cell adhesion tests, we found that carbon nanotube inhibited Schwann cell attachment on PGC nanofibrous and film matrices. However, the proliferation rates of Schwann cells were higher when they were immobilized on PGC nanofibrous matrices compared to PGC film matrices. Using western blot analysis, we found that NRG1 and P0 protein expression levels were higher for cells immobilized on PGC nanofibrous matrices compared to PG nanofibrous matrices. However, the carbon nanotube inhibited NRG1 and P0 protein expression in cells immobilized on PGC film matrices. Moreover, the NRG1 and P0 protein expression levels were higher for cells immobilized on PGC nanofibrous matrices compared to PGC film matrices. We found that the matrix topography and composition influenced Schwann cell behavior.

A composite scaffold of MSC affinity peptide-modified demineralized bone matrix particles and chitosan hydrogel for cartilage regeneration

Science.gov (United States)

Meng, Qingyang; Man, Zhentao; Dai, Linghui; Huang, Hongjie; Zhang, Xin; Hu, Xiaoqing; Shao, Zhenxing; Zhu, Jingxian; Zhang, Jiying; Fu, Xin; Duan, Xiaoning; Ao, Yingfang

2015-12-01

Articular cartilage injury is still a significant challenge because of the poor intrinsic healing potential of cartilage. Stem cell-based tissue engineering is a promising technique for cartilage repair. As cartilage defects are usually irregular in clinical settings, scaffolds with moldability that can fill any shape of cartilage defects and closely integrate with the host cartilage are desirable. In this study, we constructed a composite scaffold combining mesenchymal stem cells (MSCs) E7 affinity peptide-modified demineralized bone matrix (DBM) particles and chitosan (CS) hydrogel for cartilage engineering. This solid-supported composite scaffold exhibited appropriate porosity, which provided a 3D microenvironment that supports cell adhesion and proliferation. Cell proliferation and DNA content analysis indicated that the DBM-E7/CS scaffold promoted better rat bone marrow-derived MSCs (BMMSCs) survival than the CS or DBM/CS groups. Meanwhile, the DBM-E7/CS scaffold increased matrix production and improved chondrogenic differentiation ability of BMMSCs in vitro. Furthermore, after implantation in vivo for four weeks, compared to those in control groups, the regenerated issue in the DBM-E7/CS group exhibited translucent and superior cartilage-like structures, as indicated by gross observation, histological examination, and assessment of matrix staining. Overall, the functional composite scaffold of DBM-E7/CS is a promising option for repairing irregularly shaped cartilage defects.

Gelatin Functionalization of Biomaterial Surfaces: Strategies for Immobilization and Visualization

Directory of Open Access Journals (Sweden)

Peter Dubruel

2011-01-01

Full Text Available In the present work, the immobilization of gelatin as biopolymer on two types of implantable biomaterials, polyimide and titanium, was compared. Both materials are known for their biocompatibility while lacking cell-interactive behavior. For both materials, a pre-functionalization step was required to enable gelatin immobilization. For the polyimide foils, a reactive succinimidyl ester was introduced first on the surface, followed by covalent grafting of gelatin. For the titanium material, methacrylate groups were first introduced on the Ti surface through a silanization reaction. The applied functionalities enabled the subsequent immobilization of methacrylamide modified gelatin. Both surface modified materials were characterized in depth using atomic force microscopy, static contact angle measurements, confocal fluorescence microscopy, attenuated total reflection infrared spectroscopy and X-ray photo-electron spectroscopy. The results indicated that the strategies elaborated for both material classes are suitable to apply stable gelatin coatings. Interestingly, depending on the material class studied, not all surface analysis techniques are applicable.

Effect of Fabrication Process Parameters on the Size of Gelatin/Nanohydroxyapatite Microspheres

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S. Bagheri-Khoulenjani

2009-12-01

Full Text Available Nano-hydroxyapatite/gelatin (nHA/Ge microspheres are currently used in bone tissue engineering as bone filler. In this  study, the effect of fabrication process parameters on the particle size of nano-hydroxyapatite/gelatinmicrospheres was investigated. The nHA/Ge microspheres were fabricated using water in oil emulsion. In order to design an experimental design, a surface response model with 2 factors including the rate of shaking and water to oil volume ratio in 3 levels was applied. Particle size was evaluated by using an optical microscope. The morphology of microspheres and distribution of nano-particles within the microspheres were studied by using scanning electron microscope and Ca elemental map obtained from energy dispersive X-ray analysis (EDX, respectively. Statistical analysis of the results obtained from particle size measurements revealed that the rate of shaking has stronger influence on the particle size of microspheres. Morphological studies showed that the fabricated microspheres were spherical with smooth surface. Ca elemental map of the microspheres showed that nano-hydroxyapatite particles distributed uniformly within the microspheres.

Evaporation-based method for preparing gelatin foams with aligned tubular pore structures

Energy Technology Data Exchange (ETDEWEB)

Frazier, Shane D.; Srubar, Wil V., E-mail: wsrubar@colorado.edu

2016-05-01

Gelatin-based foams with aligned tubular pore structures were prepared via liquid-to-gas vaporization of tightly bound water in dehydrated gelatin hydrogels. This study elucidates the mechanism of the foaming process by investigating the secondary (i.e., helical) structure, molecular interactions, and water content of gelatin films before and after foaming using X-ray diffraction, Fourier transform infrared (FTIR) spectroscopy, differential scanning calorimetry and thermogravimetric analysis (TGA), respectively. Experimental data from gelatin samples prepared at various gelatin-to-water concentrations (5–30 wt.%) substantiate that resulting foam structures are similar in pore diameter (approximately 350 μm), shape, and density (0.05–0.22 g/cm{sup 3}) to those fabricated using conventional methods (e.g., freeze-drying). Helical structures were identified in the films but were not evident in the foamed samples after vaporization (~ 150 °C), suggesting that the primary foaming mechanism is governed by the vaporization of water that is tightly bound in secondary structures (i.e., helices, β-turns, β-sheets) that are present in dehydrated gelatin films. FTIR and TGA data show that the foaming process leads to more disorder and reduced hydrogen bonding to hydroxyl groups in gelatin and that no thermal degradation of gelatin occurs before or after foaming. - Highlights: • A new method is presented for fabricating gelatin foams with aligned, tubular pores. • Gelatin hydrogels were dehydrated then heated to 150 °C to induce foaming. • Vaporization of tightly (vs. loosely) bound water is the primary foaming mechanism • Foaming induced no thermal degradation but caused disorder in secondary structures • Foam microstructures are similar to those prepared using conventional methods.

Evaporation-based method for preparing gelatin foams with aligned tubular pore structures

International Nuclear Information System (INIS)

Frazier, Shane D.; Srubar, Wil V.

2016-01-01

Gelatin-based foams with aligned tubular pore structures were prepared via liquid-to-gas vaporization of tightly bound water in dehydrated gelatin hydrogels. This study elucidates the mechanism of the foaming process by investigating the secondary (i.e., helical) structure, molecular interactions, and water content of gelatin films before and after foaming using X-ray diffraction, Fourier transform infrared (FTIR) spectroscopy, differential scanning calorimetry and thermogravimetric analysis (TGA), respectively. Experimental data from gelatin samples prepared at various gelatin-to-water concentrations (5–30 wt.%) substantiate that resulting foam structures are similar in pore diameter (approximately 350 μm), shape, and density (0.05–0.22 g/cm"3) to those fabricated using conventional methods (e.g., freeze-drying). Helical structures were identified in the films but were not evident in the foamed samples after vaporization (~ 150 °C), suggesting that the primary foaming mechanism is governed by the vaporization of water that is tightly bound in secondary structures (i.e., helices, β-turns, β-sheets) that are present in dehydrated gelatin films. FTIR and TGA data show that the foaming process leads to more disorder and reduced hydrogen bonding to hydroxyl groups in gelatin and that no thermal degradation of gelatin occurs before or after foaming. - Highlights: • A new method is presented for fabricating gelatin foams with aligned, tubular pores. • Gelatin hydrogels were dehydrated then heated to 150 °C to induce foaming. • Vaporization of tightly (vs. loosely) bound water is the primary foaming mechanism • Foaming induced no thermal degradation but caused disorder in secondary structures • Foam microstructures are similar to those prepared using conventional methods.

PENGARUH JENIS LARUTAN PERENDAM SERTA METODE PENGERINGAN TERHADAP SIFAT FISIK, KIMIA, DAN FUNGSIONAL GELATIN DARI KULIT CUCUT [The Influence of Solvent Variety and Drying Method on Physical, Chemical and Functional Characteristic of Shark Skin Gelatin

Directory of Open Access Journals (Sweden)

Made Astawan 1

2003-04-01

Full Text Available Gelatin is a denaturation product of collagen and has been widely utilized for foods, photography uses, medical materials, and culture materials for microorganisms. Recently, gelatin’s uses has expanded to new applications such as health foods. Gelatin of land animal origin such as bovine and porcine has been mainly used. However, gelatin with new properties is desired to develop expanded applications. In this research gelatin was made from shark skin. The shark skin collagen was distended by acetic acid solution (acid process and sodium hydroxide solution (alkali process, and gelatin was then extracted at 80oC of water. The extracted gelatin was dried by vacuum drying and freeze drying to obtain powder product. Shark gelatin produced by the combination of alkali and vacuum drying process (OVB gelatin had better qualities on yield, water content, gel strength, melting point, and emulsion stability as compared to the others. Sensory evaluation indicated that shark gelatin had lower values in color, odor, and performance as compared to the commercial gelatin of fish, pig and bovine (SKW Biosystem. However, as compared to the commercial gelatins, OVB gelatin had a similar values on gel strength (202 bloom and emulsion stability (100%, but higher values on viscosity (8 cP and pH (9.3. Those values indicated that ray skin gelatin can be used to substitute the commercial gelatin in some food industries

Abnormal bone formation induced by implantation of osteosarcoma-derived bone-inducing substance in the X-linked hypophosphatemic mouse

International Nuclear Information System (INIS)

Yoshikawa, H.; Masuhara, K.; Takaoka, K.; Ono, K.; Tanaka, H.; Seino, Y.

1985-01-01

The X-linked hypophosphatemic mouse (Hyp) has been proposed as a model for the human familial hypophosphatemia (the most common form of vitamin D-resistant rickets). An osteosarcoma-derived bone-inducing substance was subcutaneously implanted into the Hyp mouse. The implant was consistently replaced by cartilage tissue at 2 weeks after implantation. The cartilage matrix seemed to be normal, according to the histological examination, and 35sulphur ( 35 S) uptake was also normal. Up to 4 weeks after implantation the cartilage matrix was completely replaced by unmineralized bone matrix and hematopoietic bone marrow. Osteoid tissue arising from the implantation of bone inducing substance in the Hyp mouse showed no radiologic or histologic sign of calcification. These findings suggest that the abnormalities of endochondral ossification in the Hyp mouse might be characterized by the failure of mineralization in cartilage and bone matrix. Analysis of the effects of bone-inducing substance on the Hyp mouse may help to give greater insight into the mechanism and treatment of human familial hypophosphatemia

Viscoelastic shock wave in ballistic gelatin behind soft body armor.

Science.gov (United States)

Liu, Li; Fan, Yurun; Li, Wei

2014-06-01

Ballistic gelatins are widely used as a surrogate of biological tissue in blunt trauma tests. Non-penetration impact tests of handgun bullets on the 10wt% ballistic gelatin block behind soft armor were carried out in which a high-speed camera recorded the crater׳s movement and pressure sensors imbedded in the gelatin block recorded the pressure waves at different locations. The observed shock wave attenuation indicates the necessity of considering the gelatin׳s viscoelasticity. A three-element viscoelastic constitutive model was adopted, in which the relevant parameters were obtained via fitting the damping free oscillations at the beginning of the creep-mode of rheological measurement, and by examining the data of published split Hopkinson pressure bar (SHPB) experiments. The viscoelastic model is determined by a retardation time of 5.5×10(-5)s for high oscillation frequencies and a stress relaxation time of 2.0-4.5×10(-7)s for shock wave attenuation. Using the characteristic-line method and the spherical wave assumption, the propagation of impact pressure wave front and the subsequent unloading profile can be simulated using the experimental velocity boundary condition. The established viscoelastic model considerably improves the prediction of shock wave attenuation in the ballistic gelatin. Copyright © 2014 Elsevier Ltd. All rights reserved.

Macrophage involvement affects matrix stiffness-related influences on cell osteogenesis under three-dimensional culture conditions.

Science.gov (United States)

He, Xiao-Tao; Wu, Rui-Xin; Xu, Xin-Yue; Wang, Jia; Yin, Yuan; Chen, Fa-Ming

2018-04-15

Accumulating evidence indicates that the physicochemical properties of biomaterials exert profound influences on stem cell fate decisions. However, matrix-based regulation selected through in vitro analyses based on a given cell population do not genuinely reflect the in vivo conditions, in which multiple cell types are involved and interact dynamically. This study constitutes the first investigation of how macrophages (Mφs) in stiffness-tunable transglutaminase cross-linked gelatin (TG-gel) affect the osteogenesis of bone marrow-derived mesenchymal stem cells (BMMSCs). When a single cell type was cultured, low-stiffness TG-gels promoted BMMSC proliferation, whereas high-stiffness TG-gels supported cell osteogenic differentiation. However, Mφs in high-stiffness TG-gels were more likely to polarize toward the pro-inflammatory M1 phenotype. Using either conditioned medium (CM)-based incubation or Transwell-based co-culture, we found that Mφs encapsulated in the low-stiffness matrix exerted a positive effect on the osteogenesis of co-cultured BMMSCs. Conversely, Mφs in high-stiffness TG-gels negatively affected cell osteogenic differentiation. When both cell types were cultured in the same TG-gel type and placed into the Transwell system, the stiffness-related influences of Mφs on BMMSCs were significantly altered; both the low- and high-stiffness matrix induced similar levels of BMMSC osteogenesis. Although the best material parameter for synergistically affecting Mφs and BMMSCs remains unknown, our data suggest that Mφ involvement in the co-culture system alters previously identified material-related influences on BMMSCs, such as matrix stiffness-related effects, which were identified based on a culture system involving a single cell type. Such Mφ-stem cell interactions should be considered when establishing proper matrix parameter-associated cell regulation in the development of biomimetic biomaterials for regenerative applications. The substrate stiffness

Fabrication method, structure, mechanical, and biological properties of decellularized extracellular matrix for replacement of wide bone tissue defects.

Science.gov (United States)

Anisimova, N Y; Kiselevsky, M V; Sukhorukova, I V; Shvindina, N V; Shtansky, D V

2015-09-01

The present paper was focused on the development of a new method of decellularized extracellular matrix (DECM) fabrication via a chemical treatment of a native bone tissue. Particular attention was paid to the influence of chemical treatment on the mechanical properties of native bones, sterility, and biological performance in vivo using the syngeneic heterotopic and orthotopic implantation models. The obtained data indicated that after a chemical decellularization treatment in 4% aqueous sodium chlorite, no noticeable signs of the erosion of compact cortical bone surface or destruction of trabeculae of spongy bone in spinal channel were observed. The histological studies showed that the chemical treatment resulted in the decellularization of both bone and cartilage tissues. The DECM samples demonstrated no signs of chemical and biological degradation in vivo. Thorough structural characterization revealed that after decellularization, the mineral frame retained its integrity with the organic phase; however clotting and destruction of organic molecules and fibers were observed. FTIR studies revealed several structural changes associated with the destruction of organic molecules, although all organic components typical of intact bone were preserved. The decellularization-induced structural changes in the collagen constituent resulted changed the deformation under compression mechanism: from the major fracture by crack propagation throughout the sample to the predominantly brittle fracture. Although the mechanical properties of radius bones subjected to decellularization were observed to degrade, the mechanical properties of ulna bones in compression and humerus bones in bending remained unchanged. The compressive strength of both the intact and decellularized ulna bones was 125-130 MPa and the flexural strength of humerus bones was 156 and 145 MPa for the intact and decellularized samples, respectively. These results open new avenues for the use of DECM samples as

Preparation of gelatin films incorporated with tea polyphenol nanoparticles for enhancing controlled-release antioxidant properties.

Science.gov (United States)

Liu, Fei; Antoniou, John; Li, Yue; Yi, Jiang; Yokoyama, Wallace; Ma, Jianguo; Zhong, Fang

2015-04-22

Gelatin films incorporated with chitosan nanoparticles in various free/encapsulated tea polyphenol (TP) ratios were prepared in order to investigate the influence of different ratios on the physicochemical and antioxidant properties of films. The TP-containing nanoparticles were prepared by cross-linking chitosan hydrochloride (CSH) with sulfobutyl ether-β-cyclodextrin sodium (SBE-β-CD) at three different encapsulation efficiencies (EE; ∼50%, ∼80%, and ∼100%) of TP. The stability of TP-loaded nanoparticles was maintained during the film drying process from the analysis of free TP content in the redissolved film solutions. Composite films showed no significant difference in visual aspects, while the light transmittance (250-550 nm) was decreased with incorporation of TP. Nanoparticles appeared to be homogeneously dispersed within the film matrix by microstructure analysis (SEM and AFM). TP-loaded films had ferric reducing and DPPH radical scavenging power that corresponded to the EEs. Sunflower oil packaged in bags made of gelatin films embedded with nanoparticles of 80% EE showed the best oxidation inhibitory effect, followed by 100% EE, 50% EE, and free TP, over 6 weeks of storage. However, when the gelatin film was placed over the headspace and was not in contact with the oil, the free TP showed the best effect. The results indicate that sustained release of TP in the contacting surface can ensure the protective effects, which vary with free/encapsulated mass ratios, thus improving antioxidant activities instead of increasing the dosage.

Engineering a morphogenetically active hydrogel for bioprinting of bioartificial tissue derived from human osteoblast-like SaOS-2 cells.

Science.gov (United States)

Neufurth, Meik; Wang, Xiaohong; Schröder, Heinz C; Feng, Qingling; Diehl-Seifert, Bärbel; Ziebart, Thomas; Steffen, Renate; Wang, Shunfeng; Müller, Werner E G

2014-10-01

Sodium alginate hydrogel, stabilized with gelatin, is a suitable, biologically inert matrix that can be used for encapsulating and 3D bioprinting of bone-related SaOS-2 cells. However, the cells, embedded in this matrix, remain in a non-proliferating state. Here we show that addition of an overlay onto the bioprinted alginate/gelatine/SaOS-2 cell scaffold, consisting of agarose and the calcium salt of polyphosphate [polyP·Ca(2+)-complex], resulted in a marked increase in cell proliferation. In the presence of 100 μm polyP·Ca(2+)-complex, the cells proliferate with a generation time of approximately 47-55 h. In addition, the hardness of the alginate/gelatin hydrogel substantially increases in the presence of the polymer. The reduced Young's modulus for the alginate/gelatin hydrogel is approximately 13-14 kPa, and this value drops to approximately 0.5 kPa after incubation of the cell containing scaffolds for 5 d. In the presence of 100 μm polyP·Ca(2+)-complex, the reduced Young's modulus increases to about 22 kPa. The hardness of the polyP·Ca(2+)-complex containing hydrogel remains essentially constant if cells are absent in the matrix, but it drops to 3.2 kPa after a 5 d incubation period in the presence of SaOS-2 cells, indicating that polyP·Ca(2+)-complex becomes metabolized, degraded, by the cells. The alginate/gelatine-agarose system with polyP·Ca(2+)-complex cause a significant increase in the mineralization of the cells. SEM analyses revealed that the morphology of the mineral nodules formed on the surface of the cells embedded in the alginate/gelatin hydrogel do not significantly differ from the nodules on cells growing in monolayer cultures. The newly developed technique, using cells encapsulated into an alginate/gelatin hydrogel and a secondary layer containing the morphogenetically active, growth promoting polymer polyP·Ca(2+)-complex opens new possibilities for the application of 3D bioprinting in bone tissue engineering. Copyright �

Polycaprolactone diacrylate crosslinked biodegradable semi-interpenetrating networks of polyacrylamide and gelatin for controlled drug delivery

International Nuclear Information System (INIS)

Jaiswal, Maneesh; Koul, Veena; Dinda, Amit K; Gupta, Asheesh

2010-01-01

A biodegradable semi-interpenetrating hydrogel network (semi-IPN) of polyacrylamide and gelatin was prepared using polycaprolactone diacrylate (mol. wt ∼ 640) as a crosslinker. The drug-polymer interaction and IPN formation were investigated by attenuated total reflectance-Fourier transform infrared (ATR-FTIR) and thermal gravimetric analysis (TGA). Scanning electron micrographs of lyophilized matrices revealed porous internal structure with varying pore sizes under equilibrium hydrated conditions, depending upon formulation composition. pH-dependent swelling and degradation was enhanced with increasing gelatin content and decreasing crosslinker concentration (Cs). Compression modulus (CM) (at 20% strain) increased significantly from 23 ± 1.4 to 75 ± 2.7 kPa (p 0 C). Fitting of drug release data in the Korsmeyer-Peppas model suggested sustained release behavior up to 10 days with a combination of diffusion and erosion mechanism (0.5 t /M ∞ ≤ 0.6). The newly developed porous, biodegradable and elastic semi-IPNs may serve as an ideal matrix for controlled drug release and wound healing applications. The possibilities can be explored for pharmaceutical and tissue engineering applications.

Gelatin behaviour in dilute aqueous solution : designing a nanoparticulate formulation

OpenAIRE

Farrugia, Claude; Groves, Michael J.

1999-01-01

Although it has been claimed that nanoparticles can be produced from gelatin, a naturally occurring polypeptide, the commercial conversion of animal collagen to gelatin results in a heterogeneous product with a wide molecular-weight range. This is probably responsible for the widely observed variation in the experimental conditions required for nanoparticle formation. In this study, 0.2% w/v aqueous B225 gelatin solutions were incubated under various conditions of time, temperature, pH an...

Biomineralization of Fucoidan-Peptide Blends and Their Potential Applications in Bone Tissue Regeneration

Directory of Open Access Journals (Sweden)

Harrison T. Pajovich

2017-09-01

Full Text Available Fucoidan (Fuc, a natural polysaccharide derived from brown seaweed algae, and gelatin (Gel were conjugated to form a template for preparation of biomimetic scaffolds for potential applications in bone tissue regeneration. To the Fuc–Gel we then incorporated the peptide sequence MTNYDEAAMAIASLN (MTN derived from the E-F hand domain, known for its calcium binding properties. To mimic the components of the extracellular matrix of bone tissue, the Fuc–Gel–MTN assemblies were incubated in simulated body fluid (SBF to induce biomineralization, resulting in the formation of β-tricalcium phosphate, and hydroxyapatite (HAp. The formed Fuc–Gel–MTN–beta–TCP/HAP scaffolds were found to display an average Young’s Modulus value of 0.32 GPa (n = 5 with an average surface roughness of 91 nm. Rheological studies show that the biomineralized scaffold exhibited higher storage and loss modulus compared to the composites formed before biomineralization. Thermal phase changes were studied through DSC and TGA analysis. XRD and EDS analyses indicated a biphasic mixture of β-tricalcium phosphate and hydroxyapatite and the composition of the scaffold. The scaffold promoted cell proliferation, differentiation and displayed actin stress fibers indicating the formation of cell-scaffold matrices in the presence of MT3C3-E1 mouse preosteoblasts. Osteogenesis and mineralization were found to increase with Fuc–Gel–MTN–beta–TCP/HAP scaffolds. Thus, we have developed a novel scaffold for possible applications in bone tissue engineering.

Hydrophilic PCU scaffolds prepared by grafting PEGMA and immobilizing gelatin to enhance cell adhesion and proliferation

Energy Technology Data Exchange (ETDEWEB)

Shi, Changcan; Yuan, Wenjie; Khan, Musammir; Li, Qian [School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072 (China); Feng, Yakai, E-mail: yakaifeng@tju.edu.cn [School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072 (China); Key Laboratory of Systems Bioengineering of Ministry of Education, Tianjin University, Tianjin 300072 (China); Tianjin University-Helmholtz-Zentrum Geesthacht, Joint Laboratory for Biomaterials and Regenerative Medicine, Tianjin 300072 (China); Collaborative Innovation Center of Chemical Science and Chemical Engineering (Tianjin) Tianjin 300072 (China); Yao, Fanglian [School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072 (China); Key Laboratory of Systems Bioengineering of Ministry of Education, Tianjin University, Tianjin 300072 (China); Tianjin University-Helmholtz-Zentrum Geesthacht, Joint Laboratory for Biomaterials and Regenerative Medicine, Tianjin 300072 (China); Zhang, Wencheng, E-mail: wenchengzhang@yahoo.com [Department of Physiology and Pathophysiology, Logistics University of Chinese People' s Armed Police Force, Tianjin 300162 (China)

2015-05-01

Gelatin contains many functional motifs which can modulate cell specific adhesion, so we modified polycarbonate urethane (PCU) scaffold surface by immobilization of gelatin. PCU-g-gelatin scaffolds were prepared by direct immobilizing gelatins onto the surface of aminated PCU scaffolds. To increase the immobilization amount of gelatin, poly(ethylene glycol) methacrylate (PEGMA) was grafted onto PCU scaffolds by surface initiated atom transfer radical polymerization. Then, following amination and immobilization, PCU-g-PEGMA-g-gelatin scaffolds were obtained. Both modified scaffolds were characterized by chemical and biological methods. After immobilization of gelatin, the microfiber surface became rough, but the original morphology of scaffolds was maintained successfully. PCU-g-PEGMA-g-gelatin scaffolds were more hydrophilic than PCU-g-gelatin scaffolds. Because hydrophilic PEGMA and gelatin were grafted and immobilized onto the surface, the PCU-g-PEGMA-g-gelatin scaffolds showed low platelet adhesion, perfect anti-hemolytic activity and excellent cell growth and proliferation capacity. It could be envisioned that PCU-g-PEGMA-g-gelatin scaffolds might have potential applications in tissue engineering artificial scaffolds. - Graphical abstract: PCU-g-gelatin scaffolds were prepared by direct immobilizing gelatin onto the surface of aminated PCU scaffolds (method a). To increase the immobilization amount of gelatin, PEGMAs were grafted onto the scaffold surface by SI-ATRP. PCU-g-PEGMA-g-gelatin scaffolds were prepared by method b. The gelatin modified scaffolds exhibited high hydrophilicity, low platelet adhesion, perfect anti-hemolytic activity, and excellent cell adhesion and proliferation capacity. They might have potential applications as tissue engineering scaffolds for artificial blood vessels. - Highlights: • Hydrophilic scaffolds were prepared by grafting PEGMA and immobilization of gelatins. • Grafting PEGMA enhanced the immobilization amount of gelatin

Enhancement of recovery of Neisseria meningitidis by gelatin in blood culture media.

OpenAIRE

Pai, C H; Sorger, S

1981-01-01

The efficacy of gelatin for the recovery of Neisseria meningitidis from blood cultures was evaluated in a clinical setting. The organism was isolated from seven patients with meningococcal infections in blood culture media containing 1% gelatin. In contrast, only two blood cultures from these patients were positive in media without gelatin (P less than 0.05). Gelatin did not influence the recovery of other organisms isolated during this study. Conventional blood culture media may be supplemen...

Influence of hyperbaric oxygen on biomechanics and structural bone matrix in type 1 diabetes mellitus rats.

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Pedro Henrique Justino Oliveira Limirio

Full Text Available The aim of this study was to evaluate the biomechanics and structural bone matrix in diabetic rats subjected to hyperbaric oxygen therapy (HBO.Twenty-four male rats were divided into the following groups: Control; Control + HBO; Diabetic, and Diabetic + HBO. Diabetes was induced with streptozotocin (STZ in the diabetic Groups. After 30 days, HBO was performed every 48h in HBO groups and all animals were euthanized 60 days after diabetic induction. The femur was submitted to a biomechanical (maximum strength, energy-to-failure and stiffness and Attenuated Total Reflectance Fourier transform infrared (ATR-FTIR analyses (crosslink ratio, crystallinity index, matrix-to-mineral ratio: Amide I + II/Hydroxyapatite (M:MI and Amide III + Collagen/HA (M:MIII.In biomechanical analysis, diabetic animals showed lower values of maximum strength, energy and stiffness than non-diabetic animals. However, structural strength and stiffness were increased in groups with HBO compared with non-HBO. ATR-FTIR analysis showed decreased collagen maturity in the ratio of crosslink peaks in diabetic compared with the other groups. The bone from the diabetic groups showed decreased crystallinity compared with non-diabetic groups. M:MI showed no statistical difference between groups. However, M:MIII showed an increased matrix mineral ratio in diabetic+HBO and control+HBO compared with control and diabetic groups. Correlations between mechanical and ATR-FTIR analyses showed significant positive correlation between collagen maturity and stiffness.Diabetes decreased collagen maturation and the mineral deposition process, thus reducing biomechanical properties. Moreover, the study showed that HBO improved crosslink maturation and increased maximum strength and stiffness in the femur of T1DM animals.

Influence of hyperbaric oxygen on biomechanics and structural bone matrix in type 1 diabetes mellitus rats.

Science.gov (United States)

Limirio, Pedro Henrique Justino Oliveira; da Rocha Junior, Huberth Alexandre; Morais, Richarlisson Borges de; Hiraki, Karen Renata Nakamura; Balbi, Ana Paula Coelho; Soares, Priscilla Barbosa Ferreira; Dechichi, Paula

2018-01-01

The aim of this study was to evaluate the biomechanics and structural bone matrix in diabetic rats subjected to hyperbaric oxygen therapy (HBO). Twenty-four male rats were divided into the following groups: Control; Control + HBO; Diabetic, and Diabetic + HBO. Diabetes was induced with streptozotocin (STZ) in the diabetic Groups. After 30 days, HBO was performed every 48h in HBO groups and all animals were euthanized 60 days after diabetic induction. The femur was submitted to a biomechanical (maximum strength, energy-to-failure and stiffness) and Attenuated Total Reflectance Fourier transform infrared (ATR-FTIR) analyses (crosslink ratio, crystallinity index, matrix-to-mineral ratio: Amide I + II/Hydroxyapatite (M:MI) and Amide III + Collagen/HA (M:MIII)). In biomechanical analysis, diabetic animals showed lower values of maximum strength, energy and stiffness than non-diabetic animals. However, structural strength and stiffness were increased in groups with HBO compared with non-HBO. ATR-FTIR analysis showed decreased collagen maturity in the ratio of crosslink peaks in diabetic compared with the other groups. The bone from the diabetic groups showed decreased crystallinity compared with non-diabetic groups. M:MI showed no statistical difference between groups. However, M:MIII showed an increased matrix mineral ratio in diabetic+HBO and control+HBO compared with control and diabetic groups. Correlations between mechanical and ATR-FTIR analyses showed significant positive correlation between collagen maturity and stiffness. Diabetes decreased collagen maturation and the mineral deposition process, thus reducing biomechanical properties. Moreover, the study showed that HBO improved crosslink maturation and increased maximum strength and stiffness in the femur of T1DM animals.

Autogenous bone versus deproteinised bovine bone matrix in 1-stage lateral sinus floor elevation in the severely atrophied maxilla: a randomised controlled trial.

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Merli, Mauro; Moscatelli, Marco; Mariotti, Giorgia; Rotundo, Roberto; Nieri, Michele

2013-01-01

To compare 100% deproteinised bovine bone matrix (DBBM) grafts (test group) with 100% autogenous bone (AB) grafts (control group) for lateral maxillary sinus floor elevation in a parallel group, superiority, randomised controlled trial. Patients with 1 to 3 mm of residual bone height below the maxillary sinus were randomised for sinus floor elevation with DBBM and AB grafts and simultaneous implant placement. Randomisation was computer generated with allocation concealment by sealed envelopes and the radiographic examiner was blinded to group assignment. The abutment connection was performed 8 months after surgery and insertion of the provisional prostheses was performed 9 months after surgery. Outcome variables were implant failures, prosthetic failures, complications, chair time, postoperative pain and radiographic bone level 6 months after loading. Forty patients were randomised: 20 (32 implants) to the DBBM group and 20 (27 implants) to the AB group. One patient from the AB group dropped out. Two implant failures occurred in the DBBM group and no implant failure occurred in the AB group (P = 0.4872). All of the planned prostheses could be delivered. One complication occurred in the DBBM group and 2 in the AB group (P = 0.6050). Chair time was shorter for the DBBM group, with a difference of 27.3 minutes (P = 0.0428). Pain difference measured with a visual analogue scale for 6 days post-surgery was 0.2 in favour of the DBBM group (P = 0.6838). The difference in vertical bone height was 0.0 mm (95% CI -1.1, 1.1; P = 0.9703) and the difference in marginal bone level was 0.3 in favour of AB (95% CI -0.3, 0.9; P = 0.3220). No differences apart from chair time were observed when comparing DBBM and AB grafts with simultaneous implant placement in sinus elevation.

Analyzing the cellular contribution of bone marrow to fracture healing using bone marrow transplantation in mice

International Nuclear Information System (INIS)

Colnot, C.; Huang, S.; Helms, J.

2006-01-01

The bone marrow is believed to play important roles during fracture healing such as providing progenitor cells for inflammation, matrix remodeling, and cartilage and bone formation. Given the complex nature of bone repair, it remains difficult to distinguish the contributions of various cell types. Here we describe a mouse model based on bone marrow transplantation and genetic labeling to track cells originating from bone marrow during fracture healing. Following lethal irradiation and engraftment of bone marrow expressing the LacZ transgene constitutively, wild type mice underwent tibial fracture. Donor bone marrow-derived cells, which originated from the hematopoietic compartment, did not participate in the chondrogenic and osteogenic lineages during fracture healing. Instead, the donor bone marrow contributed to inflammatory and bone resorbing cells. This model can be exploited in the future to investigate the role of inflammation and matrix remodeling during bone repair, independent from osteogenesis and chondrogenesis