CAJ | 학술논문

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.
목적 구건경니평교련대서탈세포척수지가,탐토경니평교련개성대대서탈세포척수지가항매해능력、생물역학성능급세포독성적영향. 방법 채용화학췌취법제비대서탈세포척수지가,재용5 g/L경니평용액진행화학교련개성.채용HE염색급소묘전경관찰미교련화경니평교련대서탈세포척수지가적미관결구,측량기질공경대소.분별검측교련전후대서탈세포척수지가적공극솔、함수솔이급재2.5 g/L이매용액중적실중솔.재Instron생물역학측시의상검측정상대서흉단척수、미교련급경니평교련대서탈세포척수지가적겁한견장응력화탄성모량.장대서골수간충질간세포분별재미교련화경니평교련대서탈세포척수지가침제액중배양,채용MTT법검측세포상대생장솔,평개지가적세포독성. 결과 미교련화경니평교련대서탈세포척수지가옹유상사적삼유망상공극결구,공경대소약30 μm,공극솔초과80％,량조간차이무통계학의의(P＞0.05)；경니평교련지가적함수솔위(229.7±12.5)％,현저저우미교련지가적(283.4±11.2)％(P＜0.05)；경니평교련대서탈세포척수지가재이매용액중각시상점적실중솔명현저우미교련지가(P＜0.05),생물역학측시기겁한견장응력화탄성모량칙현저증강(P＜0.05)；미교련화경니평교련대서탈세포척수지가균미관찰도명현적세포독성. 결론 경니평교련대서탈세포척수지가옹유여미교련지가상사적결구,단생물역학성능화항매해능력명현증강,차무명현세포독성,재척수손상수복적조직공정연구중구유량호적응용전경.
Objective To construct genipin-crosslinked rat acellular spinal cord scaffolds and evaluate their enzymatic degradation rate,biomechanical properties and cytotoxicity.Methods Rat spinal cord scaffolds were decellularized by chemical extraction and chemically crosslinked with 5 g/L genipin solution.Micro-structure of the uncrosslinked and genipin-crosslinked acellular spinal cord scaffolds were observed by HE staining and scanning electron microscopy and properties of pore size,porosity,water 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 acellular spinal cord scaffolds were determined on Instron mechanical testing instrument.Rat bone marrow mesenchymal stem cells were cultured in lixivium of uncrosslinked and genipin-crosslinked acellular spinal cord scaffolds and MTT 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 μm and a porosity over 80％,but there was a statistical difference between the two groups(P ＞ 0.05).Water ratio of genipincrosslinked scaffolds was (229.7 ± 12.5) ％,far lower than (283.4 ± 11.2) ％ of uncrosslinked scaffolds (P ＜ O.05) ; genipin-crosslinked acellular spinal cord scaffolds had lower weight loss at each time point than the uncrosslinked acellular 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 genipin-crosslinking (P ＜ 0.05).Furthermore,no obvious cytotoxicity was observed in the uncroslinked and genipin-crosslinked scaffolds.Conclusions Rat acellular spinal cord scaffolds present no obvious change in structure after genipin-crosslinking,but there is significant improvement in the biomechanical properties and ability against enzymatic degradation and no marked cytotoxicity.Hence,the genipincrosslinked scaffolds are promising in tissue engineering for spinal injury.