目的：本研究通过观察经地塞米松处理前后培养的人小梁细胞的变化,探讨激素性青光眼房水排出阻力增加的机制。方法：将人类小梁细胞培养至接近体内的高分化状态，用地塞米松处理后,观察细胞形态的变化，并检测以下蛋白的分布和表达：(1)myocilin/TIGR蛋白；(2)纤连蛋白；(3)肌动蛋白交联网的形成；(4)血清淀粉样物质A蛋白。结果：人小梁细胞经地塞米松处理后(1)胞体变大，排列不规则，边界模糊，呈“融合”状；(2)细胞内外myocilin/TIGR蛋白表达均明显增加，其胞外表达与纤连蛋白位置一致,提示相互作用；(3)纤连蛋白表达增加；(4)CLANs形成，细胞间连接增强；(5)血清淀粉样物质A蛋白表达增加。结论：人类小梁细胞经地塞米松处理后形态发生变化，可能与应力纤维变化及myocilin蛋白在细胞内积聚有关。细胞间边界不清，与细胞外基质蛋白过度表达、沉积有关。小梁细胞外基质沉积以及异常的细胞间连接等改变与房水排出阻力增加的病理过程有关。
Decellularized tissue engineering scaffolds appear to have the properties of similar structure and mechanical characteristics to native tissues，good biocompatibility，suitability for cell adhesion，growth and angiogenesis induction，and non-immunogenicity. Genipin has anti-inflammatory，antithrombotic and antioxidative features which can considerably suppress vascular and endothelial inflammatory activation，increase mechanical strength of biological scaffolds，inhibit inflammatory response and decrease degradation rate of biological scaffolds. By cross-linking with decellularized matrices，Genipin can further improve corresponding performance of tissue engineering matrices，which is very helpful to promote the application of tissue engineering into clinical practice of cardiothoracic surgery. This review focuses on recent research process and possible prospects of Genipin cross-linking in tissue engineering in the field of cardiothoracic surgery.
Abstract: Objective To observe the physical characteristics of decellularized porcine pulmonary valved conduits crosslinked by carbodiimide (EDC). Methods [WTBZ]Twenty porcine pulmonary valved arteries were mobilized on relative asepsis condition. They were cut longitudinally into three samples at the junction position of pulmonary valve (every sample was comprised of a part of the pulmonary conduit wall and the corresponding valve). The samples were randomly divided into three groups by lotdrawing method. Group A was the control group which was made up of the fresh porcine arterial valved conduit samples without any other treatments. Group B was comprised of porcine pulmonary samples decellularized by trypsindetergent digestion. Group Cincluded the decellularized porcine pulmonary samples crosslinked by EDC. We observed the water content, thickness, tensile strength, and shrinkage temperature of all the samples, based on which the physical characeteristics of these samples were analyzed. Results [WTBZ]Complete cellfree-pulmonary conduit matrix was achieved by trypsindetergent digestion. Compared with group A, in group B, the water content of pulmonary wall was significantly higher (P=0.000), while the water content of pulmonary valve was not significantly different; the thickness of pulmonary wall and valve (P=0.000，0.000) and tensile strength of pulmonary wall and valve (Plt;0.01） was significantly lower, while shrinkage temperature was not significantly different. Compared with group B, in group C, the water content of pulmonary wall was significantly lower (P=0.000), while the water content of pulmonary valve, and the thickness of pulmonary wall and valve were not significantly different; the tensile strength of pulmonary wall (Plt;0.01) and valve (P=0.000), and the shrinkage temperature of them (P=0.000, 0.000) were significantly higher. Compared with group A, in group C, the water content of pulmonary wall and valve, and the tensile strength of them were not statistically different, while the thickness of pulmonary wall and valve was significantly lower (P=0.000, 0.000), and the shrinkage temperature of them was significantly higher (P=0.000, 0.000). Conclusion [WTBZ]EDC crosslinking method is available for treating decellularized porcine pulmonary valved conduits in order to enhance its tensile strength, and decrease water content of pulmonary wall.
Abstract: Objective Using Amplex red fluorometric assay to detect the lysyl oxidase (LOX) enzyme activity in tissue engineered heart valve (TEHV). Methods Porcine aortic valves were decellularized with trypsin+ethylene diaminetetraacetic acid(EDTA), TritonX-100, and RNaseⅠ＋DNaseⅠ, then they were seeded by myo-fibroblasts that harvested from rats. Then they were fed with Dulbecco’s modified Eagle medium (DMEM) which contained high glucose for 27 days, they were fed with phenol red-free and serumfree DMEM for 24 hours, and the medium was harvested and used for LOX enzyme activity assays with the Amplex red fluorometric assay. And reverse transcription-polymerase chain reaction (RT-PCR) technique was used to analyze the expression of LOXmRNA in TEHV. Results All the samples produced measurable amounts of active LOX enzyme. The fluorescence units were 45.60±1.66, and the corresponding concentration of LOX enzyme were 0.123±0.003μg/ml. At the same time, all the samples expressed LOXmRNA. The expression of LOXmRNA was corresponding to the results of the Amplex red fluorometric assay. Conclusion It is feasible to detect the LOX enzyme activity in TEHV with the Amplex red fluorometric assay. And this assay gives a way to reflect that LOX plays an important role in collagen cross-linking of extracellar matrix in TEHV.
Objective To compare the effectiveness and linear wear rates between high cross-linked polyethylene cup liner and traditional polyethylene cup liner. Methods A retrospective analysis was made on the clinical data of 61 patients (64 hips) who underwent primary total hip arthroplasty (THA) between January 2005 and March 2007. According to the type of cup liner, 61 patients (64 hips) were divided into 2 groups: high cross-linked polyethylene cup liner was used in 30 cases (31 hips, trial group), and traditional polyethylene cup liner in 31 cases (33 hips, control group). There was no significant difference in gender, age, body weight, etiological factor, and preoperative Harris score between 2 groups (P gt; 0.05). Complication was observed after operation; Harris score was used to assess hip function; X-ray film analysis was done to observe osteolysis and prosthesis loosening, and to measure cumulative penetration of the femoral head and linear wear rate. Results All incisions healed by first intention without complication. All patients were followed up. The mean follow-up was 6.3 years (range, 5-7 years) in trial group, and was 6.5 years (range, 4-7 years) in control group. There was no significant difference in Harris score between 2 groups at the 3 months after operation and last follow-up (P gt; 0.05). Osteolysis was observed in 3 cases (3 hips) of control group, but no osteolysis in trial group. X-ray films showed no aseptic loosening in the patients of 2 groups. The mean cumulative penetration of trial group were significantly lower than those of control group at 1-7 years after operation (P lt; 0.05). Linear wear rate of trial group [(0.025 ± 0.002) mm/year] was significantly lower than that of control group [(0.086 ± 0.005 ) mm/year] (Z=6.804, P=0.000). Conclusion High cross-linked polyethylene cup liner has the similar effectiveness as traditional polyethylene cup liner, but it has lower linear wear rate than the traditional polyethylene cup liner.
Objective To review the application of genipin for the modification of natural biomaterials as a crosslinking agent and progress in research. Methods Domestic and foreign literature on application of genipin for the modification of natural biomaterials as a crosslinking agent was thoroughly reviewed. Results Genipin is an effective natural crosslinking agent with a very low level of cytotoxicity compared with conventional synthetic crosslinking agents. Tissues fixed with genipin can maintain a high level of stability as well as resistance to enzymatic degradation. Conclusion Genipin is a promising substitute for conventional synthetic crosslinking agents, which has offered an alternative for modification of natural biomaterials for tissue engineering.
Objective To study the release properties of basic fibroblast growth factor (bFGF) chitosan microspheres prepared by cross-linking-emulsion method using chitosan as a carrier material so as to lay a foundation for further study. Methods Using 0.6% sodium tripolyphosphate solution as a crosslinking agent and 1.5% solution of chitosan as a carrier material, bFGF chitosan microspheres were prepared by cross-linking-emulsion method. Laser particle size analyzer and Zeta electric potential analyzer were used to measure the particle diameter distribution, scanning electronic microscope to observe the morphology, and ELISA to determine the drug loading, the encapsulation rate, and the drug release properties. Results The particle size of bFGF chitosan microspheres ranged 20.312-24.152 μm. The microspheres were round with a smooth surface and uniform distribution, and it had no apparent porosity. The drug loading and encapsulation rate of microspheres were (7.57 ± 0.34) mg/g and 95.14% ± 1.58%, respectively. The bFGF chitosan microspheres could continuously release bFGF for 24 days; the bFGF level increased gradually with time and reached (820.45 ± 21.34) ng/mL at 24 days; and the microspheres had a burst effect, and the burst rate was 18.08%, and the accumulative release rate of the microspheres reached 82.05% during 24 days. Conclusion It is easy-to-operate to prepare the bFGF chitosan microspheres with the cross-linking-emulsion method. The bFGF chitosan microspheres have smooth surface, uniform distribution, and no apparent porosity.
【Abstract】 Objective To compare the properties of collagen membranes before and after crossl inked and to establ ish the foundation of appl ication of collagen membranes. Methods Fresh bovine tendons were separated and collagen was extracted by washing, smashing and acetic acid dissolving. The collagen protein was determined by ultraviolet spectrophotometer and its characteristics were analyzed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE), wavelength scanning and amino acids detecting. Collagen membranes were produced by lyophil ization. And then the biocharacteristics of the membranes before and after glutaraldehyde crossl inked were compared. BMSCs separated from volunteer’s bone marrow were seeded on collagen membranes before and after crossl inked by 2×103 in 100 μL medium, seven days after culture, the absorption spectrum of BMSCs was examined, and BMSCs were observed by scanning electron microscope (SEM). Results The contents of collagen protein were 2 mg/mL. The maximum absorption wave length appeared at about 230 nm. SDS-PAGE suggested that molecularweight of main bands was more than 66.2×103, the same as collagen marker from calf skin. There were 21.47% glycine, 12.04% pral ine and 10.18% hydroxyprol ine. No tryptophan was found. Before crossl inked, collagen membranes were in shape of white sponges and with big holes and the range of pH value was from 4.5 to 5.0. SEM showed reticular conformation and pore structure of collagen membranes, but the bore diameter was bigger. Their water-absorbing capacity was 61 times as much as their weight. The mechanical strength was 210 g/cm3. The dissolution time of collagenase was 90 minutes. After crossl inked, collagen membranes became thin, colorless, semi-transparent and compact with better tenacity. Under SEM, compact collagen fiber appeared reticular. There was lower water-absorbing capacity and pH value ranged from 6.5 to 7.0. The mechanical strength was 3 400 g/cm3 and the dissolution time of collagenase became longer. BMSCs could grow better either on before-crossl inked collagen membranes or on after-crossl inked ones. Conclusion As biomaterial scaffolds, after crossl inked collagen membranes were better than before-crossl inked ones.
Objective To review the recent advances of cross-linking reagent for producing hyaluronic acid(HA) derivative so as to provide more advice for thedevelopment of HA reagent. Methods Recent original articles related to the species, characteristic, cross-linking methodology and mechanism of the cross-linking reagent to producing HA derivative were summarized and systematically analyzed. Results The derivatives after special kinds of reagents modification would remain their own good biocompatibility and change their original rheololgical characterization and obtain relative long organism residence time. Conclusion Development of hyaluronic derivatives may widen their medical application.
OBJECTIVE: To review the recent advances of hyaluronic acid and its derivatives in medical application. METHODS: Recent original articles related to hyaluronan derivatives and their medical applications were retrieved extensively. RESULTS: Hyaluronic acid and its derivatives play important roles in visosurgery, arthritis therapy, prevention of adhesion, drug delivery, soft-tissue dilation, and percutaneous embolization. CONCLUSION: Development of hyaluronan derivatives may widen their medical application.