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作 者:魏蔷薇[1] 董艳梅[1] 陈晓峰[2] 李玉莉[2] 苗国厚[2]
机构地区:[1]北京大学口腔医学院·口腔医院牙体牙髓科,北京100081 [2]华南理工大学材料科学与工程学院,国家人体组织功能重建工程技术研究中心,广州510640
出 处:《北京大学学报(医学版)》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~~
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.
分 类 号:R318.08[医药卫生—生物医学工程]
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