Quteish D, Singh G, Dolby A E
Department of Periodontology, University of Wales College of Medicine, Heath Park, Cardiff.
J Biomed Mater Res. 1990 Jun;24(6):749-60. doi: 10.1002/jbm.820240609.
Human Type I collagen was extracted from placenta using pepsin and salt fractionation. The collagen was characterized by SDS-PAG electrophoresis dispersed in acidic medium, freeze-dried, and cross-linked in an 0.25% glutaraldehyde solution pH 4.5 for 2 days. After washing for 7 days and freeze drying the resultant collagen sponge was tested with regard to mechanical, physical, enzymatic degradation properties and biological responses. The modulus of elasticity was found to be 289 +/- 10 g/mm2 and the sponge was insoluble in water, buffered saline, or tissue culture medium over a period of 6 weeks with swelling occurring at less than 5% of volume. The sponge had a high fluid binding capacity, amounting to 56 +/- 5 mL tissue culture medium per gram of dry weight. Bacterial collagenase produced slow degradation of the sponge with complete disappearance by 24 h only when high concentrations (200 units enzyme per mg of the collagen sponge) were used. Cytotoxicity studies using human gingival and periodontal ligament fibroblasts revealed less than 5% apparent cytotoxicity or proliferation. Subcutaneous implantation was followed by resorption and vascularization over a period of 6-8 weeks. It was concluded that the collagen sponge prepared from human Type I collagen has potential as a graft material in oral surgical procedures.
使用胃蛋白酶和盐分级分离法从胎盘中提取人I型胶原蛋白。通过在酸性介质中分散的SDS-PAG电泳对胶原蛋白进行表征，冷冻干燥，并在pH 4.5的0.25%戊二醛溶液中交联2天。洗涤7天后，冷冻干燥得到的胶原蛋白海绵，对其机械、物理、酶降解特性和生物学反应进行测试。发现弹性模量为289±10 g/mm2，该海绵在6周内不溶于水、缓冲盐水或组织培养基，体积膨胀小于5%。该海绵具有高液体结合能力，每克干重可达56±5 mL组织培养基。细菌胶原酶对海绵的降解缓慢，只有在使用高浓度（每毫克胶原蛋白海绵200单位酶）时，24小时后才会完全消失。使用人牙龈和牙周膜成纤维细胞进行的细胞毒性研究显示，明显的细胞毒性或增殖小于5%。皮下植入后，在6-8周内会发生吸收和血管化。得出的结论是，由人I型胶原蛋白制备的胶原蛋白海绵在口腔外科手术中作为移植材料具有潜力。
