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富血小板纤维蛋白渗出物促进人牙周膜细胞的增殖和成骨分化。

Platelet‑rich fibrin exudate promotes the proliferation and osteogenic differentiation of human periodontal ligament cells in vitro.

机构信息

Department of Stomatology, The People's Hospital of Longhua, Shenzhen, Guangdong 518109, P.R. China.

Key Laboratory of Oral Medicine, Guangzhou Institute of Oral Disease, Stomatological Hospital of Guangzhou Medical University, Guangzhou, Guangdong 510140, P.R. China.

出版信息

Mol Med Rep. 2018 Nov;18(5):4477-4485. doi: 10.3892/mmr.2018.9472. Epub 2018 Sep 10.

Abstract

The purpose of the present study was to evaluate the effects of platelet‑rich fibrin (PRF) exudate on the proliferation, osteogenic differentiation and mineralization of human periodontal ligament cells (hPDLCs) in vitro. In the present study PRF was obtained with permission, from the peripheral blood of healthy donors and PRF exudates were collected on the 7th day of incubation. hPDLCs were obtained from healthy premolars, cultured by a tissue explant method and identified with anti‑vimentin and anti‑cytokeratin antibodies. PRF exudates were added to hPDLCs in different concentrations to evaluate cell proliferation and osteogenic differentiation. The proliferation of hPDLCs was measured using a colorimetric assay. Osteogenic differentiation and mineralization were determined by Alizarin red staining, alkaline phosphatase activity (ALP), western blotting and reverse transcription‑quantitative polymerase chain reaction. Cell proliferation was enhanced by addition of the PRF exudate, which also promoted the formation of mineralized matrix nodules and upregulated ALP activity and osteoblast‑associated levels of osteocalcin, runt‑related transcription factor and osterix gene expression. As these stimulatory effects occurred in a dose‑dependent manner, it was concluded that high concentrations of the PRF exudate served an essential role in the proliferation, osteogenic differentiation and mineralization of hPDLCs in vitro. The present study demonstrated that PRF exudate enhanced hPDLC proliferation, induced the osteoblastic differentiation of hPDLCs into mineralized tissue‑formation cells in vitro, and may therefore provide potential benefits for periodontal tissue engineering; contributing to the primary processes of periodontal tissue regeneration. From the perspective of both economics and biology, PRF has greater clinical benefits than analogous growth factors.

摘要

本研究旨在评估富血小板纤维蛋白(PRF)渗出物对人牙周膜细胞(hPDLCs)体外增殖、成骨分化和矿化的影响。本研究中 PRF 是在获得健康供体外周血后获得的,并在孵育第 7 天收集 PRF 渗出物。hPDLCs 取自健康前磨牙,采用组织块培养法培养,并通过抗波形蛋白和抗细胞角蛋白抗体进行鉴定。将 PRF 渗出物以不同浓度添加到 hPDLCs 中,以评估细胞增殖和成骨分化。通过比色法测量 hPDLC 增殖。通过茜素红染色、碱性磷酸酶活性(ALP)、western blot 和逆转录-定量聚合酶链反应来确定成骨分化和矿化。添加 PRF 渗出物可增强 hPDLC 的增殖,促进矿化基质结节的形成,并上调 ALP 活性以及成骨细胞相关的骨钙素、 runt 相关转录因子和骨桥蛋白基因表达。由于这些刺激作用呈剂量依赖性,因此可以得出结论,高浓度的 PRF 渗出物对 hPDLC 体外增殖、成骨分化和矿化具有重要作用。本研究表明,PRF 渗出物增强了 hPDLC 的增殖,诱导 hPDLC 向矿化组织形成细胞的成骨分化,因此可能为牙周组织工程提供潜在益处;有助于牙周组织再生的初始过程。从经济和生物学的角度来看,PRF 比类似的生长因子具有更大的临床益处。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7693/6172397/6575991eb9f1/MMR-18-05-4477-g00.jpg

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