摘要
目的:犬自体皮下非受力部位植入犬骨髓基质干细胞(bonemarrowstromalcells,BMSCs)复合珊瑚羟基磷灰石(corallinehydroxyapatite,CHA),观察成骨情况及其转归。方法:体外分离培养、成骨诱导、扩增犬BMSCs,并通过细胞化学、免疫细胞化学检测成骨表型。将第2代细胞接种于CHA上,形成细胞-材料复合物,植于9只犬右侧腹部皮下,并在对称部位植入CHA作为对照。术后4、12、26周取材,HE染色,观察新生组织结构;通过形态计量学分析,对成骨情况量化,并行t检验。结果:成骨诱导后,细胞碱性磷酸酶染色阳性,免疫细胞化学显示骨钙蛋白表达阳性。HE染色示植入4周后,无明显骨形成;12周后,实验组有较多新生骨小梁形成,苦味酸-硫堇和Masson染色示骨基质和胶原形成良好;26周后,实验组骨小梁量有减少趋势。图像分析显示,实验组12周成骨量最高,差异有统计学意义。结论:犬BMSCs诱导后具有成骨活性,复合CHA后,可促进形成组织工程化骨。在无应力刺激情况下,随时间延长,局部成骨活动减弱。
PURPOSE： This study was to investigate long term osteogenesis of canine bone marrow stromal cells （BMSCs） complex with coralline hydroxyapatite implanted subcutaneously. METHODS： Isolated BMSCs were expanded and osteogenically induced in vitro. Their osteogenic phenotype was evaluated by immunocytochemistry and cytochemistry. In 9 dogs, cell-scaffold constructs made from autologous induced BMSCs （P2） and coralline hydroxyapatite（CHA） were implanted into right abdomen subcutaneously as experimental group. CHA was implanted into contralateral side as control group. The engineered bones were harvested 4,12,26 weeks post-implantation and processed for HE staining, then evaluated by histomorphometry and Student t test. RESULTS： In induced BMSCs, immunocytochemistry showed positive staining of osteocalcin. Cytochemistu＇ also showed alkaline phosphatase （AKP） activity. Histologically, there was no bone formation at 4 weeks.HE staining showed newly formed trabeculae at 12 weeks, bone matrix and collagen formation were also demonstrated by Picric-Thionine and Masson staining. Histomorphometry showed more bone trabecula was observed in the experimental group than in the control group at 12 and 26 weeks. However, less bone was formed at 26 weeks when compared to that at 12 weeks in the experimental group. CONCLUSIONS： CHA loaded with osteogeneically induced BMSCs can form tissue engineered bone in canine subcutaneously, the local ossification decreased without stress stimulation with time going.
出处
《中国口腔颌面外科杂志》
CAS
2005年第3期207-211,共5页
China Journal of Oral and Maxillofacial Surgery
基金
国家"973"组织工程基本科学问题项目(G1999054308)
国家"863"组织工程化骨构建技术研究与产品开发(2002AA205011)
上海市教育委员会"重中之重"重点学科资助项目
