京尼平交联大鼠脱细胞脊髓支架的制备及其生物力学特性研究  

Preparation and biomechanical property of genipin-crosslinked rat acellular spinal cord scaffolds

作　　者：蒋涛[1] 任先军[1] 阴洪[1] 王开见[1] 周长立[1] 田永阳[1] 

机构地区：[1]第三军医大学附属新桥医院骨科,重庆400037

出　　处：《中华创伤杂志》2014年第2期180-184,共5页Chinese Journal of Trauma

基　　金：国家青年基金资助项目（31000438）;国家自然基金资助项目（81271362）

摘　　要：目的 构建京尼平交联大鼠脱细胞脊髓支架,探讨京尼平交联改性对大鼠脱细胞脊髓支架抗酶解能力、生物力学性能及细胞毒性的影响. 方法 采用化学萃取法制备大鼠脱细胞脊髓支架,再用5 g/L京尼平溶液进行化学交联改性.采用HE染色及扫描电镜观察未交联和京尼平交联大鼠脱细胞脊髓支架的微观结构,测量基质孔径大小.分别检测交联前后大鼠脱细胞脊髓支架的孔隙率、含水率以及在2.5 g/L胰酶溶液中的失重率.在Instron生物力学测试仪上检测正常大鼠胸段脊髓、未交联及京尼平交联大鼠脱细胞脊髓支架的极限牵张应力和弹性模量.将大鼠骨髓间充质干细胞分别在未交联和京尼平交联大鼠脱细胞脊髓支架浸提液中培养,采用MTT法检测细胞相对生长率,评价支架的细胞毒性. 结果 未交联和京尼平交联大鼠脱细胞脊髓支架拥有相似的三维网状孔隙结构,孔径大小约30 μm,孔隙率超过80％,两组间差异无统计学意义（P＞0.05）；京尼平交联支架的含水率为（229.7±12.5）％,显著低于未交联支架的（283.4±11.2）％（P＜0.05）；京尼平交联大鼠脱细胞脊髓支架在胰酶溶液中各时相点的失重率明显低于未交联支架（P＜0.05）,生物力学测试其极限牵张应力和弹性模量则显著增强（P＜0.05）；未交联和京尼平交联大鼠脱细胞脊髓支架均未观察到明显的细胞毒性. 结论 京尼平交联大鼠脱细胞脊髓支架拥有与未交联支架相似的结构,但生物力学性能和抗酶解能力明显增强,且无明显细胞毒性,在脊髓损伤修复的组织工程研究中具有良好的应用前景.Objective To construct genipin-crosslinked rat acellular spinal cord scaffolds and e- valuate their enzymatic degradation rate, biomeehanical properties and cytotoxieity. Methods Rat spi- nal cord scaffolds were deeellularized by chemical extraction and chemically crosslinked with 5 g/＇L genip- in solution. Micro-structure of the unerosslinked and genipin-erosslinked acellular spinal cord scaffolds were observed by HE staining and scanning electron microscopy and properties of pore size, porosity, wa- ter ratio, and degradation rate in 2.5 g/L trypsin enzyme solution were examined. Ultimate tensile strength and elastic modulus of normal rat thoracic spinal cord, uncrosslinked and genipin-crosslinked a- cellular spinal cord scaffolds were determined on Instron mechanical testing instrument. Rat bone marrow mesenehymal stem cells were cultured in lixivium of uncrosslinked and genipin-crosslinked acellular spinal cord scaffolds and Mq＇T assay for relative cell growth rate was test to evaluate the cytotoxicity of scaffolds. Results The uncrosslinked and the genipin-crosslinked acellular spinal cord scaffolds possessed a similar three-dimensional mesh-porous structure with a mean pore diameter about 30 txm and a porosity over 80%, but there was a statistical difference between the two groups （ P 〉 0.05 ）. Water ratio of genipin- crosslinked scaffolds was （229.7 -＋ 12.5 ） %, far lower than （283.4 -＋ 11.2） % of unerosslinked scaffolds （ P 〈 0.05 ） ; genipin-crosslinked aeellular spinal cord scaffolds had lower weight loss at each time point than the uncrosslinked aeellular spinal cord scaffolds （ P 〈 0.05 ）, but the stability in trypsin, ultimate tensile strength and elastic modulus of acellular spinal cord scaffolds were significantly enhanced by genip- in-crosslinking （ P 〈 0.05 ）. Furthermore, no obvious eytotoxicity was observed in the uncroslinked and genipin-crosslinked scaffolds. Conclusions Rat acellular spinal cord scaffolds present no obvious change in structure after ge

关 键 词：组织支架 生物力学 脱细胞脊髓 

分 类 号：R318.08[医药卫生—生物医学工程]