京尼平交联脱细胞纤维环基质/壳聚糖水凝胶与兔纤维环源干细胞的生物相容性  

Biocompatibility of rabbit annulus fibrosus-derived stem cells with genipin cross-linked decellularized annulus fibrosus matrix/chitosan hydrogels

作　　者：刘晨[1] 赵泉来[1] 王凌挺[1] 王弘[1] 刘平[1] 李斌[2] 徐宏光[1] 

机构地区：[1]皖南医学院弋矶山医院脊柱外科,安徽省芜湖市241001 [2]苏州大学附属第一医院骨科,江苏省苏州市215007

出　　处：《中国组织工程研究》2016年第21期3143-3149,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81272048);安徽省自然科学基金(1308085MH152)~~

摘　　要：背景:为改善脱细胞基质的力学性能及降解不可控性,前期研究制备了京尼平交联脱细胞纤维环基质/壳聚糖水凝胶纤维环组织工程支架。目的:观察京尼平交联脱细胞纤维环基质/壳聚糖水凝胶和纤维环源干细胞的生物相容性,以及京尼平交联脱细胞纤维环基质/壳聚糖水凝胶的体内降解情况。方法:制备京尼平交联脱细胞纤维环基质/壳聚糖水凝胶,通过扫描电镜和接触角测试仪观察水凝胶内部结构及表面亲水性。将第1代兔纤维环源干细胞种植于京尼平交联脱细胞纤维环基质/壳聚糖水凝胶上3 d,行细胞骨架染色,通过倒置免疫荧光显微镜和扫描电镜观察细胞形态,并绘制细胞生长曲线。将京尼平交联脱细胞纤维环基质/壳聚糖水凝胶种植于新西兰大耳白兔肌间隙内,4周后观察水凝胶降解情况及与周围组织炎症反应。结果与结论:京尼平交联脱细胞纤维环基质/壳聚糖水凝胶支架为多孔网状结构,接触角为(39.94±1.61)°;纤维环源干细胞黏附于京尼平交联脱细胞纤维环基质/壳聚糖水凝胶支架表面,增殖较快,生长状态良好,说明京尼平交联脱细胞纤维环基质/壳聚糖水凝胶具有良好的生物相容性。种植于兔体内的水凝胶支架在4周后有所降解,周围肌肉组织有炎症细胞存在。BACKGROUND： To improve the mechanical properties and uncontrollability of degradation of decellularized matrix, we manufactured genipin cross-linked decellularized annulus fibrosus matrix/chitosan hydrogels as annulus fibrosus tissue-engineered scaffold. OBJECTIVE： To observe the biocompatibility of annulus fibrosus-derived stem cells with genipin cross-linked decellularized annulus fibrosus matrix/chitosan hydrogels, and the degradation of hydrogels in vivo. METHODS： Genipin cross-linked decellularized annulus fibrosus matrix/chitosan hydrogel scaffolds were prepared, and the internal structure and hydrophilic property of hydrogels was detected by scanning electron microscopy and contact angle meter, respectively. Afterwards, passage 1 rabbit annulus fibrosus-derived stem cells were seeded on the scaffolds for 3 days, cell morphology was observed by inverted immunofluorescence microscopy and scanning electron microscopy after cytoskeleton staining, and additionally, curve of cell growth was obtained. In the meanwhile, the scafflods were transplanted into the intermuscular space of New Zealand white rabbits, and the degradation and inflammatory reaction of the hydrogels were observed after 4 weeks. RESULTS AND CONCLUSION： The prepared hydrogel scaffold showed porous and network structure and the contact angle was（39.94±1.61）°; annulus fibrosus-derived stem cells adhered well on the scaffold surface with a faster growth rate. Furthermore, at 4 weeks after transplantation, the hydrogel in vivo was degraded, and inflammatory cells appeared. These findings indicate that genipin cross-linked decellularized annulus fibrosus matrix/chitosan hydrogels have good biocompatibility.

关 键 词：壳聚糖 水凝胶 干细胞 组织工程 生物材料 材料相容性 纤维环组织工程 脱细胞基质 国家自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]