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楚克伦富血小板纤维蛋白(PRF)以剂量依赖的方式刺激体外人口腔骨髓间充质干细胞的增殖和分化。

Choukroun's platelet-rich fibrin (PRF) stimulates in vitro proliferation and differentiation of human oral bone mesenchymal stem cell in a dose-dependent way.

机构信息

Department of Biomaterials, Institute for Clinical Sciences, The Sahlgrenska Academy at University of Gothenburg, Sweden.

出版信息

Arch Oral Biol. 2010 Mar;55(3):185-94. doi: 10.1016/j.archoralbio.2010.01.004. Epub 2010 Feb 21.

Abstract

BACKGROUND

Choukroun's platelet-rich fibrin (PRF) is an autologous leukocyte- and platelet-rich fibrin biomaterial. The purpose of this study was to analyse the in vitro effects of PRF on human bone mesenchymal stem cells (BMSC), harvested in the oral cavity after preimplant endosteal stimulation.

MATERIALS AND METHODS

BMSCs from primary cultures were cultivated with or without a PRF membrane originating from the same donor as for the cells, in proliferation or osteoblastic differentiation conditions. After 7 days, the PRF membranes were removed. A series of cultures were performed using 2 PRF membranes, in order to measure the dose-dependent effect. Cell counts, cytotoxicity tests, alkaline phosphatase (ALP) activity quantification, Von Kossa staining and mineralisation nodules counts were performed at 3, 7, 14, 21 and 28 days. A last independent series was carried on up to 14 days, for a morphological scanning electron microscope (SEM) observation.

RESULTS

PRF generated a significant stimulation of the BMSC proliferation and differentiation throughout the experimental period. This effect was dose-dependent during the first weeks in normal conditions, and during the whole experimentation in differentiation conditions. The cultures without PRF in differentiation conditions did not rise above the degree of differentiation of the cultures in normal conditions with 1 or 2 PRF up to the 14th and 28th day, respectively. The SEM culture analysis at day 14 allowed to show the mineralisation nodules which were more numerous and more structured in the groups with PRF compared to the control groups.

DISCUSSION AND CONCLUSIONS

This double contradictory proliferation/differentiation result may be due to the numerous components of PRF, particularly the presence of leukocytes: any culture with PRF is in fact a coculture with leukocytes. It could be the source of differential geographic regulation processes within the culture. The combination of oral BMSC and PRF might offer many potential clinical and biotechnological applications, and deserves new studies.

摘要

背景

Choukroun 富血小板纤维蛋白(PRF)是一种自体白细胞和血小板丰富的纤维蛋白生物材料。本研究旨在分析 PRF 对口腔内预植入骨内刺激后采集的人骨髓间充质干细胞(BMSC)的体外影响。

材料和方法

从原代培养物中培养 BMSC,在增殖或成骨分化条件下,有无来自同一供体的 PRF 膜。7 天后,去除 PRF 膜。为了测量剂量依赖性效应,进行了一系列使用 2 个 PRF 膜的培养。细胞计数、细胞毒性试验、碱性磷酸酶(ALP)活性定量、Von Kossa 染色和矿化结节计数在 3、7、14、21 和 28 天进行。最后一组独立实验持续到 14 天,进行形态学扫描电子显微镜(SEM)观察。

结果

PRF 在整个实验期间显著刺激 BMSC 的增殖和分化。在正常条件下的前几周,这种作用是剂量依赖性的,而在分化条件下则是整个实验期间的。在分化条件下没有 PRF 的培养物在第 14 天之前,其分化程度均未超过正常条件下培养物的分化程度,分别使用 1 个或 2 个 PRF。第 14 天的 SEM 培养物分析表明,与对照组相比,PRF 组的矿化结节数量更多且更具结构。

讨论和结论

这种双重矛盾的增殖/分化结果可能是由于 PRF 的许多成分,特别是白细胞的存在:任何含有 PRF 的培养物实际上都是与白细胞的共培养物。它可能是培养物内差异地理调节过程的来源。口腔 BMSC 和 PRF 的结合可能提供许多潜在的临床和生物技术应用,值得进一步研究。

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