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.

Yuan-yuan Jia; Jing-yi Zhou; Yue Chang; Fang AN; Xiao-wei LI; Xiao-yue XU; Xiu-li Sun; Chun-yang Xiong; Jian-liu Wang

Return

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.

Author: Yuan-Yuan JIA ^{1
,

2} ; Jing-Yi ZHOU ^{1
,

2} ; Yue CHANG ^{1
,

2} ; Fang AN ^{1
,

2} ; Xiao-Wei LI ^{1
,

2} ; Xiao-Yue XU ^{1
,

2} ; Xiu-Li SUN ^{1
,

2} ; Chun-Yang XIONG ³ ; Jian-Liu WANG ^{1
,

2}
Author Information

1. Department of Obstetrics and Gynecology, Peking University People's Hospital
2. Beijing Key Laboratory of Female Pelvic Floor Disorders, Beijing 100044, China.
3. Department of Mechanics and Bioengineering, College of Engineering, Peking University, Beijing 100871, China.
Publication Type:Journal Article
Keywords: Adipose Mesenchymal Stem Cells; Basic Fibroblast Growth Factor; Epidermal Growth Factor; Fibroblasts; Tissue Engineering
MeSH: Animals; Cell Proliferation; Cells, Cultured; Collagen; metabolism; Epidermal Growth Factor; physiology; Fibroblast Growth Factor 2; physiology; Fibroblasts; physiology; Pelvic Floor; Rats; Regeneration
From: Chinese Medical Journal 2018;131(17):2089-2096
CountryChina
Language:English
Abstract: Background:Fibroblasts were the main seed cells in the studies of tissue engineering of the pelvic floor ligament. Basic fibroblast 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 fibroblasts.
Methods:Fibroblasts were differentiated from rat adipose mesenchymal stem cells (ADSCs). Flow cytometry and immunohistochemistry were used for cell identification. 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 identified 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 fibroblasts 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 fibroblasts displayed significant 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 fibroblasts was 10 ng/ml at which fibroblasts 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.