摘要
目的:观察人牙髓细胞(human dental pulp cells,hDPCs)在生物活性支架上的增殖及分化情况。方法:采用酶消化法培养hDPCs,传至第4代用于实验。用免疫组织化学法鉴定细胞并测定细胞基质前体细胞抗原-1(stro-mal precursor antigen-1,STRO-1)表达率。实验使用3种支架,包括胶原(collagen,COL)支架、胶原-生物活性玻璃(collagen-bioglass,COL-BG)支架及胶原-生物活性玻璃-聚己内酯(collagen-bioglass-polycaprolacton,COL-BG-PCL)支架。将hDPCs植入支架,采用MTT法于6 h、1 d、3 d、5 d、7 d、14 d和21 d测定hDPCs增殖,在14 d进行碱性磷酸酶染色。结果:实验所用hDPCs中含有STRO-1阳性细胞;hDPCs在COL-BG支架及COL-BG-PCL支架上的增殖显著高于COL支架(P<0.05),尤其在14 d和21 d COL-BG支架及COL-BG-PCL支架的光密度值是COL支架的5倍;COL-BG支架及COL-BG-PCL支架上的碱性磷酸酶染色区明显较COL支架广泛。结论:hDPCs在COL-BG支架及COL-BG-PCL支架上增殖和分化活跃,优于传统的COL支架。
Objective：To investigate the proliferation and differentiation of the human dental pulp cells （hDPCs） on the bioactive scaffolds. Methods： Primary HDPCs were harvested from impacted third mo- lars of healthy adult individuals （ 18 - 25 years of age） by enzyme digestion, expanded and cultured. The cells used for this investigation were the 4 th passage. Immunohistochemical methods were used to verify that the cells were dental pulp cells. The expression of stromal precursor antigen-1 （ STRO-1 ） was determined by flow cytometry. Three different types of scaffolds were used ： collagen （ COL）, collagen / hio- glass （COL-BG） and collagen / bioglass / polyeaprolactone （COL-BG-PCL）. Cell proliferation on the scaffolds was determined using a MTT assay at hour 6, on days 1, 3, 5, 7, 14 and 21. On day 14, the scaffolds were stained with the alkaline phosphatase （ALP） staining kit. Results： The tested cells had STRO-1 positive cells. The proliferation of HDPCs was significantly higher on the COL-BG scaffold and COL-BG-PCL scaffold as compared with COL scaffold （P 〈 0.05）. EspeciaUy on days 14 and 21, the optical density value of hioglass composite scaffold were 5 times that of the COL scaffold. The ALP positive staining area was observed more extensively on the COL-BG scaffold and COL-BG-PCL scaffold than on the COL scaffold. Conclusion： As comparison with the COL scaffold, HDPCs＇ proliferation and dif- ferentiation oresent more activity on the COL-BG and COL-BG-PCL scaffolds.
出处
《北京大学学报（医学版）》
CAS
CSCD
北大核心
2013年第3期484-488,共5页
Journal of Peking University:Health Sciences
基金
国家自然科学基金(50830101
51072055)资助~~
关键词
牙髓
组织支架
胶原
生物活性玻璃
细胞增殖
Dental pulp
Tissue scaffolds
Collagen
Bioglass
Cell proliferation
