兔脂肪源性干细胞的成骨诱导及其与异种松质骨的生物相容性研究

Osteogenic-induction of Rabbit Adipose-derived Stem Cells and the Study of the Biocompatibility between the Cells and Heterogeneity Cancellous Bone

探讨异种松质骨作为支架材料的前景,并观察其与成骨诱导前后脂肪源性干细胞的生物相容性。取兔脂肪源性干细胞,诱导成骨并检测。将诱导成骨和未诱导的脂肪源性干细胞接种至松质骨支架上。扫描电镜观察,并计算细胞24 h的贴壁率。将细胞-支架复合物及单纯支架分别植入家兔皮下,10天后处死取材,HE染色行组织学分析。结果表明:兔脂肪源性干细胞在体外诱导培养后,碱性磷酸酶含量升高,诱导2周后茜素红染色阳性,提示脂肪源性干细胞向成骨方向分化。扫描电镜观察显示:诱导成骨和未诱导的脂肪源性干细胞均能与异种松质骨支架材料复合良好,二者24 h的贴壁率差别无统计学意义。由此说明:脂肪源性干细胞能够在体外诱导成骨,且异种松质骨支架材料与其有良好的生物相容性,提示脂肪源性干细胞能够作为骨组织工程的种子细胞,同时也提示异种松质骨能够作为支架材料用于骨组织工程研究。

To isolate adipose-derived stem cells（ADSC） from the adipose tissue of rabbit and induce ADSC differentiation along osteogenic lineages in vitro, and to investigate the prospect of the heterogeneity cancellous bone to be used as scaffold in bone-tissue-engineering, to observe the biocompatibility between ADSC and heterogeneity cancellous bone scaffold in vitro. ADSC was isolated, ADSC differentiation was induced along osteogenic lineages in vitro and detected. ADSC and osteogenic-induced ADSC were seeded onto cancellous bone scaffold. These cells living on the scaffold were observed through scanning electron microscope and the rate of adherence of the cells in 24 hours was calculated. The cell-scaffold compounds were implanted and scaffolded merely subcutaneously, the sample was harvested 10 days later after the rabbits were executed. Histological analysis was made through HE staining. Results showed that ADSC isolated from rabbit could be induced differentiation along osteogenic lineages. The alkaline phosphatase contained in osteogenic-induced ADSC was more than that contained in non-induced ADSC. Alizarin red staining was positive when ADSC were osteogenic-induced for 14 days. These results showed that ADSC were induced differentiation along osteogenic lineages. Scanning electron microscope showed that both non-induced ADSC and osteogenic-induced ADSC could compound well with heterogeneity cancellous bone, the difference between the adherence rates of the two kinds of cell in 24 hours was not statistically significant. It proves that ADSC can be induced differentiation along osteogenic lineages in vitro, and can live well on heterogeneity cancellous bone scaffold. ADSC can be used as seed cells in bone-tissue-engineering, and that heterogeneity cancellous bone can be used as scaffold in bone-tissue-engineering.