[交联前后胶原膜性能分析]
[Analysis of properties of collagen membranes before and after crosslinked].
作者信息
Chen Lin, Lv Yang, Guan Lidong, Wang Yunfang, He Lijuan, Li Yanhua, Bai Cixian, Liu Daqing, Pei Xuetao
机构信息
Institute of Transfusion Medicine, Academy of Military Medical Sciences, Beijing, 100850, P.R. China.
出版信息
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2008 Feb;22(2):183-7.
OBJECTIVE
To compare the properties of collagen membranes before and after crosslinked and to establish the foundation of application 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 lyophilization. And then the biocharacteristics of the membranes before and after glutaraldehyde crosslinked were compared. BMSCs separated from volunteer's bone marrow were seeded on collagen membranes before and after crosslinked by 2 x 10(3) in 100 microL 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 molecular weight of main bands was more than 66.2 x 10(3), the same as collagen marker from calf skin. There were 21.47% glycine, 12.04% praline and 10.18% hydroxyproline. No tryptophan was found. Before crosslinked, 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-crosslinked collagen membranes or on after-crosslinked ones.
CONCLUSION
As biomaterial scaffolds, after crosslinked collagen membranes were better than before-crosslinked ones.
目的
比较交联前后胶原膜的性能，为胶原膜的应用奠定基础。
方法
分离新鲜牛肌腱，经洗涤、粉碎、乙酸溶解提取胶原蛋白。用紫外分光光度计测定胶原蛋白含量，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）、波长扫描和氨基酸检测分析其特性。采用冻干法制备胶原膜。然后比较戊二醛交联前后胶原膜的生物学特性。将从志愿者骨髓中分离的骨髓间充质干细胞（BMSCs）以2×10³个细胞/100μL培养基接种于交联前后的胶原膜上，培养7天后，检测BMSCs的吸收光谱，并用扫描电子显微镜（SEM）观察BMSCs。
结果
胶原蛋白含量为2mg/mL。最大吸收波长出现在约230nm处。SDS-PAGE显示主带分子量大于66.2×10³，与小牛皮肤胶原蛋白标志物相同。甘氨酸含量为21.47%，脯氨酸含量为12.04%，羟脯氨酸含量为10.18%。未检测到色氨酸。交联前，胶原膜呈白色海绵状，孔隙大，pH值范围为4.5至5.0。SEM显示胶原膜呈网状结构和孔隙结构，但孔径较大。其吸水能力为其重量的61倍。机械强度为210g/cm³。胶原酶溶解时间为90分钟。交联后，胶原膜变薄、无色、半透明且致密，韧性更好。在SEM下，致密的胶原纤维呈网状。吸水能力降低，pH值范围为6.5至7.0。机械强度为3400g/cm³，胶原酶溶解时间变长。BMSCs在交联前和交联后的胶原膜上均能较好生长。
结论
作为生物材料支架，交联后的胶原膜优于交联前的胶原膜。
Zotero 插件