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浓缩生长因子对人牙周膜细胞体外功能及Wnt3a表达的影响

Effect of concentrated growth factors on function and Wnt3a expression of human periodontal ligament cells in vitro.

作者信息

Qiao Jing, An Na

机构信息

a Department of Periodontology, the First Clinical Division , Peking University School and Hospital of Stomatology , Beijing , China.

b Department of General Dentistry II , Peking University School and Hospital of Stomatology , Beijing , China.

出版信息

Platelets. 2017 May;28(3):281-286. doi: 10.1080/09537104.2016.1213381. Epub 2016 Sep 6.

Abstract

Concentrated growth factors (CGFs), the new generation of platelet concentrate products, appear to exhibit superior potential for tissue regeneration. However, there are only a few studies supporting this. This study was designed to investigate the effect of CGFs on proliferation and alkaline phosphatase (ALP) activity of human periodontal ligament cells (hPDLCs) in vitro. Furthermore, as bone homeostasis is fundamentally controlled by Wnt-mediated signals, we also investigated Wnt3a expression of hPDLCs after treatment of CGFs. hPDLCs and CGFs were obtained from the same volunteer. CGFs or combination of recombined human TGF-β1 (rhTGF-β1) and PDGF-AB (rhPDGF-AB) were added to hPDLCs in different concentrations. The rate of proliferation was analyzed by an MTT assay. ALP activity was assessed using p-NPP assay. Quantitative RT-PCR was used to evaluate the gene expression of Wnt3a. In a range of concentrations, CGFs significantly promoted the proliferation of hPDLCs in a dose-dependent manner. ALP activity was also enhanced by CGFs in a dose-dependent and time-dependent manner. The stimulatory effect of CGFs was much greater than rhTGF-β1 and rhPDGF-AB combination. Quantitative RT-PCR results showed that Wnt3a mRNA expression was increased at 24 h in hPDLCs treated by CGFs. CGFs can enhance hPDLCs proliferation and ALP activity and may have great potential in clinical and biotechnological applications.

摘要

浓缩生长因子(CGFs)是新一代血小板浓缩产品,似乎展现出卓越的组织再生潜力。然而,仅有少数研究支持这一点。本研究旨在体外研究CGFs对人牙周膜细胞(hPDLCs)增殖及碱性磷酸酶(ALP)活性的影响。此外,由于骨稳态从根本上由Wnt介导的信号控制,我们还研究了CGFs处理后hPDLCs的Wnt3a表达。hPDLCs和CGFs取自同一名志愿者。将不同浓度的CGFs或重组人转化生长因子-β1(rhTGF-β1)与血小板衍生生长因子-AB(rhPDGF-AB)的组合添加到hPDLCs中。通过MTT法分析增殖率。使用对硝基苯磷酸酯(p-NPP)法评估ALP活性。采用定量逆转录聚合酶链反应(qRT-PCR)评估Wnt3a的基因表达。在一定浓度范围内,CGFs以剂量依赖性方式显著促进hPDLCs的增殖。CGFs还以剂量依赖性和时间依赖性方式增强ALP活性。CGFs的刺激作用远大于rhTGF-β1与rhPDGF-AB的组合。定量RT-PCR结果显示,CGFs处理的hPDLCs在24小时时Wnt3a mRNA表达增加。CGFs可增强hPDLCs的增殖和ALP活性,在临床和生物技术应用中可能具有巨大潜力。

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