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纤连蛋白固定化微槽钛基质对人牙龈成纤维细胞增殖及基因和蛋白质表达的影响

The Effect of Fibronectin-Immobilized Microgrooved Titanium Substrata on Cell Proliferation and Expression of Genes and Proteins in Human Gingival Fibroblasts.

作者信息

Kim Eun-Cheol, Lee Do Yun, Lee Myung-Hyun, Lee Hong Jae, Kim Kyung-Hee, Leesungbok Richard, Ahn Su-Jin, Park Su-Jung, Yoon Joon-Ho, Jee Yu-Jin, Lee Sang Cheon, Lee Suk Won

机构信息

1Department of Oral and Maxillofacial Pathology and Institute of Oral Biology, School of Dentistry, Kyung Hee University, 26 Kyungheedae-ro, Dongdaemun-gu, Seoul, 02447 Republic of Korea.

Doctorplant Dental Clinic, 885 Gyeongin-ro, Yeongdeungpo-gu, Seoul, 07303 Republic of Korea.

出版信息

Tissue Eng Regen Med. 2018 Aug 28;15(5):615-627. doi: 10.1007/s13770-018-0153-7. eCollection 2018 Oct.

Abstract

BACKGROUND

We aimed to determine the effect of fibronectin (FN)-immobilized microgrooved titanium (Ti) on human gingival fibroblast proliferation, gene expression and protein expression.

METHODS

Photolithography was used to fabricate the microgrooved Ti, and amine funtionalization (silanization) was used for FN immobilization on titanium surfaces. Cell proliferation, gene expression and protein expression were analyzed, followed by multiple regression analysis for determining the influential factors on cell proliferation.

RESULTS

FN-immobilized microgrooved Ti significantly enhanced the fibroblast proliferation in various timelines of culture, among which a burst of fivefold increase is induced at 96 h of culture compared to that on the control smooth Ti. We suggest a presence of the synergistic promotion effect of microgrooves and FN immobilization on fibroblast proliferation. Through a series of analyses on the expression of various genes and proteins involved in cell adhesion and proliferation, cyclin-dependent kinase 6, cyclin D1, integrin α5, oncogene c-Src, osteonectin, paxillin and talin-2 were determined as influential factors on promoting fibroblast proliferation induced by FN-immobilized microgrooved Ti.

CONCLUSION

FN-immobilized microgrooved Ti can act as an effective surface for enhancing fibroblast proliferation, and can be used for promoting soft tissue response on the connective tissue attachment zone of biomaterial surfaces.

摘要

背景

我们旨在确定固定有纤连蛋白(FN)的微槽钛(Ti)对人牙龈成纤维细胞增殖、基因表达和蛋白表达的影响。

方法

采用光刻法制备微槽钛,并通过胺功能化(硅烷化)将FN固定在钛表面。分析细胞增殖、基因表达和蛋白表达,随后进行多元回归分析以确定影响细胞增殖的因素。

结果

固定有FN的微槽钛在不同培养时间显著增强了成纤维细胞的增殖,与对照光滑钛相比,在培养96小时时诱导了五倍的激增。我们认为微槽和FN固定对成纤维细胞增殖存在协同促进作用。通过对参与细胞黏附和增殖的各种基因和蛋白表达的一系列分析,确定细胞周期蛋白依赖性激酶6、细胞周期蛋白D1、整合素α5、癌基因c-Src、骨连接蛋白、桩蛋白和踝蛋白-2是固定有FN的微槽钛诱导成纤维细胞增殖的影响因素。

结论

固定有FN的微槽钛可作为增强成纤维细胞增殖的有效表面,并可用于促进生物材料表面结缔组织附着区的软组织反应。

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