摘要
目的比较犬牙槽骨来源干细胞(ABSC)和骨髓间充质干细胞(BMSC)的体外成骨能力,为骨组织工程的种子细胞选择提供新思路。方法取同一只犬的牙槽骨骨组织和髂骨骨髓作为ABSC和BMSC的来源。取第3代细胞进行一系列体外检测:MTT法检测两种细胞的增殖能力;成骨诱导和常规培养条件下,不同时间点进行茜素红和碱性磷酸酶染色,观察钙盐沉积和碱性磷酸酶合成能力,同时对细胞内碱性磷酸酶活力进行半定量检测;成骨诱导后7、14、21 d利用RT-PCR法检测关键性成骨因子[Ⅰ型胶原(COL-1)、碱性磷酸酶(ALP)、骨钙素(OCN)]的表达情况。结果相较BMSC,ABSC增殖能力略高,钙结节和碱性磷酸酶的表达能力更强。COL-1早期在ABSC中的表达高于BMSC,并随着时间呈下降趋势。ALP在ABSC中的表达始终高于BMSC,且在14 d达到最高峰。OCN总体呈上升趋势,14 d和21 d有明显表达,且ABSC高于BMSC。结论犬ABSC的成骨能力与BMSC差别不大,提示犬ABSC可以成为骨组织工程优先选择的种子细胞之一。
Objective To compare the osteogenic ability of two different canine stem cells in the application of bone tissue engineering in vitro,namely alveolar bone-derived stem cells（ ABSC） and bone marrow mesenchymal stromal cells（ BMSC）. Methods ABSC and BMSC were prepared from the alveolar bone and bone marrow of a dog. The third generation of these cells were used for the following studies. Firstly,the proliferation of the two cells was detected by MTT assay. Secondly,after osteogenic induction,the osteogenic ability was detected by alizarin red staining and alkaline phosphatase staining. Thirdly,after osteogenic induction for 7,14 and 21 d,the activity of intracellular alkaline phosphatase was detected by the p NPP method. Fourthly,after osteogenic induction,the expression of collagen Ⅰ（ COL-1）,alkaline phosphatase（ ALP） and osteocalcin（ OCN） was analyzed by RT-PCR.Results ABSC showed higher proliferative ability. After osteogenic induction,ABSC produced more calcium nodules and alkaline phosphatase. The results showed that the expression of COL-1 in ABSC was higher than that in BMSC and both of them decreased gradually. The expression of ALP in them reached the highest peak at 14 d.OCN was significantly expressed in ABSC at 14 and 21 d,and was higher in BMSC. Conclusion The osteogenic abilities of ABSC are similar to that of BMSC,which is an ideal seed cell in the research of bone tissue engineering.
出处
《安徽医科大学学报》
CAS
北大核心
2016年第9期1252-1257,共6页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金(编号:31370983)
关键词
骨髓干细胞
牙槽骨干细胞
成骨分化
bone marrow mesenchymal stromal cells
alveolar bone-derived stem cells
osteogenic differentiation
