组织工程与重建外科杂志

组织工程与重建外科杂志 ›› 2012, Vol. 8 ›› Issue (2): 61-64.doi: 10.3969/j.issn.1673-0364.2012.02.001

• 论著 •    下一篇

PRP对人皮肤成纤维细胞增殖、体外合成胶原与透明质酸的影响

常毅，邓辰亮，屈悦，郑江红，万伟东，杨松林   

  1. 上海交通大学医学院附属第六人民医院整形外科
  • 发布日期:2020-07-23

The Effect of PRP on Proliferation of Human Skin Fibroblasts and Expression of Collagen and Hyaluronic Acid in Vitro

CHANG Yi,DENG Chenliang,QU Yue,ZHENG Jianghong,WAN Weidong,YANG Songlin   

  1. Department of Plastic Surgery,Shanghai Sixth People’s Hospital,Shanghai Jiaotong University School of Medicine,Shanghai 200233,China.
  • Published:2020-07-23
  • Contact: 国家自然科学基金　　（81000837）

摘要： 目的研究不同浓度的富血小板血浆(PRP)对人皮肤成纤维细胞(HSFs)增殖、体外合成胶原与透明质酸的影响。方法原代培养人皮肤成纤维细胞,用含不同浓度PRP(1%、5%、10%、20%、30%、50%)的培养液进行细胞培养,无血清培养液为阴性对照,含10%新生小牛血清的培养液为阳性对照。培养第3、5、7天使用四甲基偶氮唑盐(MTT)法测定细胞增殖率,应用RT-PCR测定胶原蛋白表达情况,ELISA测定透明质酸功能变化。结果 MTT检测发现所有不同浓度的PRP均有明显促进HSFs增殖的作用,与阴性对照组均有明显差异(P<0.01),且随着时间递增,增殖效果也明显增强;RT-PCR测定50%PRP浓度组及阴性、阳性对照组中目的基因COL1及COL3均显著表达,但组间差异不明显;ELISA测定实验组透明质酸功能均增强,与阴性对照组有明显差异(P<0.01),PRP浓度在30%～50%时表达效果明显增强。结论PRP对HSFs的增殖、合成胶原与透明质酸的功能有明显促进效果。

关键词: 富血小板血浆, 皮肤成纤维细胞, 胶原蛋白, 透明质酸

Abstract: Objective To explore the effect of platelet-rich plasma(PRP) with different concentrations on proliferation of human skin fibroblasts and expression of collagen and hyaluronic acid(HA) in vitro.Methods Primary cultured human skin fibroblasts(HSFs) with different concentrations(1%、5%、10%、20%、30%、50%) of PRP culture medium were treated as experimental groups,and take serum-free medium as negative control group and culture medium containing 10% newborn calf serum as positive control group.MTT,RT-PCR and ELISA were respectively used to measure HSFs proliferation,collagen expression and hyaluronic acid expression at the 3rd,5th and 7th day.Results MTT assay indicates that the experimental groups can all promote the proliferation of HSFs compared with the negative control group(P<0.01),and the effect showed on increasing tendency.RT-PCR detection showed that the target genes Collagen Type 1(COL1) and Collagen Type 3(COL3) were all expressed in the 50% PRP group,negative control group and positive control group,but there were no significant differences among the three groups.ELISA showed the enhancement of HA expression compared with the negative control group(P<0.01).The HA expression was significantly increased with the PRP concentration of 30%-50%(P<0.01).Conclusion PRP could promote the proliferation of HSFs and the function of the synthesis of collagen and HA.

Key words: Platelet-rich plasma, Human skin fibroblasts, Collagen, Hyaluronic acid

中图分类号: