摘要:
Objective To evaluate the biological potential of adipose-derived stromal cell(ADSCs) to porosity characteristicof biomimeticage combined neodymium-doped yttrium aluminum garne(Nd: YAG) lasewith Arg-Gly-Asp (RGD) modification.MethodSurface topography of chitosantricalcium phosphate-polycaprolactone(β-TCP/CS/PCL) matrix wamade by Nd:YAG laser, undeoscillating culture, tissue-engineered cage waconstructed in the following ways: ADSCwere loaded to β-TCP/CS/PCL cage of pore range 400-550 μm (large pore size of cage) with RGD modification (group A), goat' ADSCwere cultured to β-TCP/CS/PCL cage of pore range 200-350 μm(medium pore size of cage) with RGD modification (group B), ADSCimplantation with β-TCP/CS/PCL cage of pore range 100-150 μm (small pore size of cage) pluRGD modification (group C).Scanning eletromicroscope and confocal microscopy were used to observe cellulapattem, survival rate, cell activity, ALP and collagen type Ⅰ level were detected a1,4, 8, 12, 16, 20, 24, 28 days.OPG expression waalso assayed adiffereninterval.ResultUndeobservation of scanning eletromicroscope and confocal microscopy, new tissue showed more remarkable cell proliferation with abundanECM in group B, compared with othegroups,the survival rate in group wasignificantly highe[(74.56 ± 0.57) %, P < 0.05], level of cell activity, ALP, collagen type Ⅰ were significantly highe[(0.72 ± 3.13) AU, (73.27-± 1.54) U/L, (71.18 ± 2.51) mg/L, P < 0.05], and remarkable OPG expression waalso seen.Conclusion Topography-surfaced treatmenof β-TCP/CS/PCL cage in medium pore range enhancethe cell proliferation and actasatisfactory spinal cage candidate.%目的 评估融合器(cage)表面改性及孔隙特征与骨化脂肪基质细胞活性间关系,并验证优化改性的方法.方法 掺钕钇铝石榴石(Nd:YAG)激光处理不同孔径(400 ~ 550 μm、200~350 μm及100~ 150μm)磷酸三钙/壳聚糖/聚已内酯(β-TCP/CS/PCL) cage,精氨酸-甘氨酸-天冬氨酸(RGD)表面修饰,如下接种羊骨化脂肪基质细胞(ADSCs):共分为A组(大孔径cage +RGD表面改性+ADSCs),B组(中孔径cage+ RGD表面改性+ADSCs)和C组(小孔径cage+ RGD表面改性+ ADSCs),振荡模式下培养;第1、4、8、12、16、20、24、28天,电镜及共聚焦显微镜下观察,检测存活率、增殖、碱性磷酸酶及Ⅰ型胶原活性,分析细胞生长情况,裸鼠体内成骨测试.结果 B组细胞生长旺盛、细胞外基质丰富,存活率高于其他组[(74.56±0.57)%,P<0.05],增殖活力、碱性磷酸酶及Ⅰ型胶原水平均高于其他组[(0.72 ±3.13) AU、(73.27±1.54) U/L、(71.18 ±2.51) mg/L,P<0.05],骨保护素(OPG)表达明显.结论 中孔径cage具备稳定的细胞相容性,为理想的候选cage材料.