小型猪富血小板血浆对牙周膜干细胞的成骨诱导

Effect of miniature pig platelet-rich plasma on osteogenic induction of periodontal ligament stem cells in vitro

背景:组织工程技术为牙周炎致骨组织缺损的修复提供了新的思路。目的:探寻富血小板血浆在小型猪牙周膜干细胞成骨诱导中的作用。方法:采集贵州小型猪静脉血,三次离心制备富血小板血浆。采用组织块法分离培养小型猪牙周膜干细胞,分别将含体积分数0.8%,1.0%,1.2%的富血小板血浆与牙周膜干细胞共同培养3,7,14,21d,以未添加富血小板血浆的牙周膜干细胞作为对照。结果与结论:体积分数0.8%,1.0%,1.2%的富血小板血浆成骨诱导第14天,碱性磷酸酶活性达到峰值,其中体积分数1.0%的富血小板血浆诱导的细胞碱性磷酸酶活性最高,第21天时碱性磷酸酶活性降低。茜素红染色显示富血小板血浆成骨诱导第21天细胞染色呈阳性。单克隆纯化后细胞的生长曲线从第3天起进入对数生长期,第8天细胞数量达到顶峰。说明富血小板血浆具有诱导牙周膜干细胞成骨的能力,以体积分数1.0%时诱导成骨效率最优。

BACKGROUND： Tissue engineering technology provides a new way for bone defects caused by periodontitis. OBJECTIVE： To explore the effect of platelet-rich plasma （PRP） on osteogenic induction of miniature pig periodontal ligament stem cells （PDLSCs）. METHODS： Collected the Guizhou miniature pig’s venous blood and prepared the centrifuged PRP three times; tissue block method was used to culture the periodontal ligament cells. 0.8%, 1.0%, 1.2% concentration of PRP were co-cultured with PDLSCs for 3, 7, 14 and 21 days, respectively. No PRP addition as control group. RESULTS AND CONCLUSION： Alkaline phosphatase （AKP） activity reach a peak at the 14th day after induced with 0.8%, 1.0% and 1.2% PRP, and highest activity of AKP was induced by 1.0% PRP. AKP activity was reduced at the 21st day. Alizarin red staining showed the PDLSCs were positive at 21st day after osteogenic induction of PRP. Purified monoclonal cell growth curve entering the logarithmic growth phase since the 3rd day, the amount of cells reached a peak at the 8th day. PRP can induce osteogenic PDLSC performance, especially the 1.0% PRP induction showed the optimal efficiency in vitro.