Copolymer cell/scaffold constructs for bone tissue engineering: Co-culture of low ratios of human endothelial and osteoblast-like cells in a dynamic culture system
Copolymer cell/scaffold constructs for bone tissue engineering: Co-culture of low ratios of human endothelial and osteoblast-like cells in a dynamic culture system
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DOI：
10.1002/jbm.a.34414
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发表时间：
2013-04-01
影响因子：
4.9
通讯作者：
Mustafa, Kamal
中科院分区：
文献类型：
--
作者：
Xing, Zhe;Xue, Ying;Mustafa, Kamal
本研究的目的是比较不同比例的人脐静脉内皮细胞(HUVECs)在动态培养体系中对人成骨样细胞(HOB)和毛细血管样结构(CLS)的成骨活性的影响。将HOB和HUVEC以5：1(5：1组)和2：1(2：1组)的比例共培养成聚(L-丙交酯)-共(1，5-二氧杂环己酮)[聚(LLA-co-DXO)]支架。分别在5、15和25天后采集样本。对两组的横断面进行处理，并披露HUVEC的CLS。DsDNA法检测细胞存活率。以5：1比例接种的细胞有较好的存活率。提取总RNA，进行逆转录反应。PCR芯片显示不同比例的人脐静脉内皮细胞对多种成骨基因表达的影响不同。RT-PCR结果与PCR芯片结果一致，5：1组成骨相关基因的表达高于2：1组，特别是在第25天，碱性磷酸酶、胰岛素样生长因子1等基因的表达水平高于2：1组。酶联免疫吸附试验显示，5：1共培养的细胞在孵育25天后产生的IGF1高于2：1的细胞。结果表明，在动态培养条件下，HOB与低比例的HUVEC在共聚支架中共培养可形成CLS，并显著影响成骨标志物的表达。(C)2012年Wiley期刊，Inc.《生物材料》A部分，2013年。
The aim of this study was to compare the effect of different ratios of human umbilical vein endothelial cells (HUVECs) on osteogenic activity of human osteoblast-like cells (HOB) and capillary-like structure (CLS), seeded into copolymer scaffolds in a dynamic culture system. HOB and HUVEC were co-cultured into poly(L-lactide)-co-(1,5-dioxepan-2-one) [poly(LLA-co-DXO)] scaffolds at ratios of 5:1 (5:1 group) and 2:1 (2:1 group). Samples were collected after 5, 15, and 25 days. Cross-sections were processed and the CLS from HUVEC was disclosed in both groups. Cell viability was determined by dsDNA assay. Cells seeded at the ratio of 5:1 had good viability. Total RNA was isolated and the reverse transcription reaction was performed. The influences on the expression of several osteogenic genes were various with regarding to different ratios of HUVEC demonstrated by the PCR array. The RT-PCR results was in consistent with the PCR array results that several osteogenesis related genes had higher expression in the 5:1 group than in the 2:1 group, especially at day 25, such as alkaline phosphatase, insulin-like growth factor 1 (IGF1), and so forth. ELISA showed that the production of IGF1 after 25 days of incubation were higher in cells co-cultured at the 5:1 ratio than at the 2:1 ratio. The results show that under dynamic culture conditions, co-culture of HOB with a low ratio of HUVEC in copolymer scaffolds results in CLS formation and significantly influenced the expression of osteogenic markers. (c) 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2013.