人脂肪干细胞复合珊瑚支架在三维培养下成骨活性的初步研究

目的探讨人脂肪干细胞（ADSCs）作为种子细胞复合珊瑚支架材料在体外三维培养下的成骨活性。方法取行脂肪抽吸术患者的脂肪组织，I型胶原酶消化进行培养，贴壁细胞传代，取第2代细胞进行诱导，培养基中添加成骨诱导剂地塞米松、β-甘油磷酸钠、抗坏血酸和维生素D3。成骨诱导后复合珊瑚继续培养，以未诱导的ADSCs复合物为对照，进行生长曲线、DIO染色后的共聚焦荧光显微镜、碱性磷酸酶（AKP）和骨钙蛋白（OCN）生物化学定量检测。结果7～8d后ADSCs在材料上生长进入平台期，诱导ADSCs复合珊瑚7d后生长良好。诱导ADSCsAKP表达随着时间的推移不断增强，而且在检测的各个时间点，诱导ADSCsAKP表达水平均明显高于未诱导ADSCs（均P〈0．05）。诱导ADSCs在珊瑚支架上第7天后检测到OCN表达，并一直增高，且从第7天开始OCN表达均明显高于未诱导ADSCs（均P〈0.05）。结论脂肪干细胞复合珊瑚支架在三维培养下能够向成骨细胞表型分化。

Osteogenesis ability of human adipose stem cells on coral scaffolds in vitro

Objective To investigate the osteogenic ability of human adipose stem cells （ADSCs） in coral scaffolds in vit- ros. Methods ADSCs were isolated from liposuction aspirates, then digested by type I collagenase; the passage 2 cells were cultured in conditioned medium containing dexamethasone, β-glycerophosphate, ascorbate-2-phosphate and VD3. The in- duced cells were co-cultured with coral scaffold and non-induced ADSCs-coral construct served as control. The constructs were examined with growth curve, confocal microscopy and quantitative histochemistry. Results The growth curve demonstrat- ed that the proliferation of ADSCs on coral scaffolds reached plateau after 7-8 d of co-culture. Confocal microscopy and quanti- tative histochemistry showed that ADSCs grew well and began to express osteocalcin on the coral scaffolds after 7d of culture. Quantitative histochemistry demonstrated that the expressions of alkaline phosphatase （AKP） and osteocalcin （OCN） in induced ADSCs-scaffold were higher than those in non-induced ADSCs-scaffold （P〈0.05）. Conclusion The coral scaffold is a good biomaterial for ADSCs growth and being induced to osteoblasts.