The effects of human platelet lysate on dental pulp stem cells derived from impacted human third molars
The effects of human platelet lysate on dental pulp stem cells derived from impacted human third molars
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人血小板裂解物对人阻生第三磨牙牙髓干细胞的影响
DOI：
10.1016/j.biomaterials.2012.03.057
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发表时间：
2012-07-01
期刊：
影响因子：
14
通讯作者：
Yu, Qing
中科院分区：
文献类型：
--
作者：
Chen, Bo;Sun, Hai-Hua;Yu, Qing
人血小板裂解液（PL）被认为是胎牛血清（FBS）在牙髓干细胞（DPSCs）大规模扩增中的替代品。然而，PL对人DPSCs增殖和分化的生物学效应和最佳浓度尚不清楚。我们从取出的第三磨牙牙髓中分离和扩增干细胞，并在体外和体内评估了PL对细胞增殖能力和分化潜力的影响。在检测前，免疫细胞化学染色和基于流式细胞术的细胞分选表明，来源于人牙髓的细胞含有间充质干细胞群。细胞在组织培养塑料或羟基磷灰石-磷酸三钙（HA/TCP）生物材料上生长，并在存在或不存在各种浓度的人PL的情况下与正常或牙源性/成骨培养基孵育，以进一步研究。所有检测条件下，5% PL均能显著提高DPSCs的增殖能力（P < 0.05）。然而，当细胞在1% PL或10% PL中培养时，这种增强是不一致的；10% PL显著抑制细胞增殖，因此被排除在进一步的分化试验之外。矿物条件下碱性磷酸酶活性和钙沉积测定结果显示，5% PL培养基也显著促进了DPSCs矿化分化（P < 0.05）。扫描电镜和改良的Ponceau三色染色显示，5% PL和矿化培养基处理的细胞与HA/TCP生物材料的整合能力很强，并且在播种期14 d后以广泛的片状结构完全覆盖支架表面。此外，5% PL对两种体内移植模型的组织再生均有显著的积极作用。我们得出结论，适当浓度的PL可以增强人DPSCs在体内和体外的增殖和矿化分化，这支持在未来的临床试验中使用PL作为FBS的替代品或非人畜共患的细胞培养佐剂。然而，阐明PL产物的分子复杂性，确定决定特定干细胞命运的基本生长因子和确定剂量的标准需要进一步的研究。(C) 2012 Elsevier Ltd.版权所有。
Human platelet lysate (PL) has been suggested as a substitute for fetal bovine serum (FBS) in the large-scale expansion of dental pulp stem cells (DPSCs). However, the biological effects and the optimal concentrations of PL for the proliferation and differentiation of human DPSCs remain unexplored. We isolated and expanded stem cells from the dental pulp of extracted third molars and evaluated the effects of PL on the cells' proliferative capacity and differentiation potential in vitro and in vivo. Before testing, immunocytochemical staining and flow cytometry-based cell sorting showed that the cells derived from human dental pulp contained mesenchymal stem cell populations. Cells were grown on tissue culture plastic or on hydroxyapatite-tricalcium phosphate (HA/TCP) biomaterials and were incubated with either normal or odontogenic/osteogenic media in the presence or absence of various concentrations of human PL for further investigation. The proliferation of DPSCs was significantly increased when the cells were cultured in 5% PL under all testing conditions (P < 0.05). However, this enhancement was inconsistent when the cells were cultured in 1% PL or in 10% PL; 10% PL significantly inhibited cell proliferation and was therefore excluded from further differentiation testing. Culture medium containing 5% PL also significantly promoted the mineralized differentiation of DPSCs, as indicated by the measurement of alkaline phosphatase activity and calcium deposition under mineral-conditioned media (P < 0.05). Scanning electron microscopy and modified Ponceau trichrome staining showed that the cells treated with 5% PL and mineralizing media were highly capable of integrating with the HA/TCP biomaterials and had fully covered the surface of the scaffold with an extensive sheet-like structure 14 d after seeding. In addition, 5% PL showed significantly positive effects on tissue regeneration in two in vivo transplantation models. We conclude that the appropriate concentration of PL enhances the proliferation and mineralized differentiation of human DPSCs both in vitro and in vivo, which supports the use of PL as an alternative to FBS or a nonzoonotic adjuvant for cell culture in future clinical trials. However, the elucidation of the molecular complexity of PL products and the identification of both the essential growth factors that determine the fate of a specific stem cell and the criteria to establish dosing require further investigation. (C) 2012 Elsevier Ltd. All rights reserved.