Bone marrow mesenchymal stem cells stimulated by bFGF up-regulated protein expression in comparison with periodontal fibroblasts in vitro

被引:13
作者
Colenci, Renato [1 ]
da Silva Assuncao, Luciana Reichert [2 ]
Mogami Bomfim, Suely Regina [3 ]
Golim, Marjorie de Assis [4 ]
Deffune, Elenice [5 ]
Penha Oliveira, Sandra Helena [6 ]
机构
[1] UNESP Univ Estadual Paulista, DDS, Sch Dent, Sao Paulo, Brazil
[2] Univ Fed Parana, UFPR, Sch Dent, Dept Stornatol, BR-80060000 Curitiba, Parana, Brazil
[3] UNESP Univ Estadual Paulista, Sch Vet Med, Dept Clin Surg & Anim Reprod, Sao Paulo, Brazil
[4] UNESP Univ Estadual Paulista, Sch Med, Botucatu Blood Ctr, Lab Flow Cytometry, Sao Paulo, Brazil
[5] UNESP Univ Estadual Paulista, Sch Med, Botucatu Blood Ctr, Lab Cellular Engn, Sao Paulo, Brazil
[6] UNESP Univ Estadual Paulista, Dept Basic Sci, Sch Dent, Sao Paulo, Brazil
基金
巴西圣保罗研究基金会;
关键词
Bone marrow; Mesenchymal stem cells; Periodontal ligament; Fibroblasts; bFGF; STROMAL CELLS; GROWTH-FACTOR; LIGAMENT; DIFFERENTIATION; REGENERATION; FIBRONECTIN;
D O I
10.1016/j.archoralbio.2013.11.017
中图分类号
R78 [口腔科学];
学科分类号
1003 ;
摘要
Objective: The aim of this study was to evaluate, in vitro, the role of bFGF in the proliferation and expression of collagen type I and fibronectin of dog bone marrow mesenchymal stem cells (dBMMSCs) in comparison with the expression of the same proteins in dog periodontal fibroblasts (dPLFs). Design: dBMMSCs from the iliac crest were cultivated in Dulbecco's Modified Eagle's Medium (DMEM). Flow cytometry analysis (FCA) was used to characterize dBMMSC. Cells were stimulated with bFGF (1, 5 and 10 ng/mL) after 24 and 48 h. Real time RT-PCR was performed to verify collagen type I and fibronectin expressions. MTT assay was used to confirm cellular proliferation. Statistical analyses were performed (ANOVA and Kruskal Wallis tests; p < 0.05). Results: FCA showed 55.98% of CD34+ and 32.67% of CD90+ after bone marrow aspiration; 3.33% of CD34+ and 33.0% of CD90+ before P1. After P2, 10.54% of dBMMSCs expressed CD90, whereas after P3, this number decreased to 1.58%. dPLFs presented 4.04% of CD90+ and 1.05% of CD34+ after P3. MU evaluation showed increase in dBMSC proliferation with 5 ng/mL bFGF-stimulus after 24-h. Both collagen land fibronectin expression were very similar between the two cells groups after 24-h stimulation with 1 ng/mL bFGF concentration. Fibronectin and collagen I expressions were higher after 24-h stimulation with 5 ng/mL bFGF. Conclusion: dBMMSCs (1 ng/mL-bFGF stimulus after 24 h) are very similar to dPLFs as regards morphological and immunostaining characteristics, and collagen and/or fibronectin production. The dBMMSCs presented the highest protein expression rates with 5 ng/mL-bFGF stimulus after 24-h. (C) 2013 Elsevier Ltd. All rights reserved.
引用
收藏
页码:268 / 276
页数:9
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