Effect of Optimized Concentrations of Basic Fibroblast Growth Factor and Epidermal Growth Factor on Proliferation of Fibroblasts and Expression of Collagen: Related to Pelvic Floor Tissue Regeneration
Effect of Optimized Concentrations of Basic Fibroblast Growth Factor and Epidermal Growth Factor on Proliferation of Fibroblasts and Expression of Collagen: Related to Pelvic Floor Tissue Regeneration
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DOI：
10.4103/0366
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Wang Jian-Liu
中科院分区：
文献类型：
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作者：
Jia Yuan-Yuan;Zhou Jing-Yi;Chang Yue;An Fang;Li Xiao-Wei;Xu Xiao-Yue;Sun Xiu-Li;Xiong Chun-Yang;Wang Jian-Liu
Background: Fibroblasts were the main seed cells in the studies of tissue engineering of the pelvic floor ligament. Basic fbroblast growth factor (bFGF) and epidermal growth factor (EGF) were widely studied but at various concentrations. This study aimed to optimize the concentrations of combined bFGF and EGF by evaluating their effects on proliferation and collagen secretion of fbroblasts. Methods: Fibroblasts were differentiated from rat adipose mesenchymal stem cells (ADSCs). Flow cytometry and immunohistochemistry were used for cell identifcation. The growth factors were applied at concentrations of 0, 1, 10, and 100 ng/ml as three groups: (1) bFGF.alone, (2) EGF alone, and (3) bFGF mixed with EGF. Cell proliferation was evaluated by Cell Counting Kit 8 assays. Expression of Type I and III collagen (Col-I and Col-III) mRNAs was evaluated by real time quantitative reverse transcription polymerase chain reaction. Statistical analysis was performed with SPSS software and GraphPad Prism using one way analysis of variance and multiple t test. Results: ADSCs were successfully isolated from rat adipose tissue as identifed by expression of typical surface markers CD29, CD44, CD90, and CD45 in flow cytometry. Fibroblasts induced from ADSC, compared with ADSCs, were with higher mRNA expression levels of Col I and Col III (F = 1.29, P = 0.0390). bFGF, EGF, and the mixture of bFGF with EGF can enhanced fbroblasts proliferation, and the concentration of 10 ng/ml of the mixture of bFGF with EGF displayed most effectively (all P < 0.05). The expression levels of Col-I and Col-III mRNAs in fbroblasts displayed signifcant increases in the 10 ng/ml bFGF combined with EGF group (all P < 0.05). Conclusions: The optimal concentration of both bFGF and EGF to promote cell proliferation and collagen expression in fbroblasts was 10 ng/ml at which fbroblasts grew faster and secreted more Type I and III collagens into the extracellular matrix, which might contribute to the stability of the pelvic floor microenvironment..
