Centre for Rural Health, School of Health Sciences University of Tasmania, Newnham, Launceston, 7250, Australia.
Department of Restorative Dentistry, Faculty of Dentistry, University of Malaya, Kuala Lumpur, Malaysia.
Clin Oral Investig. 2019 Oct;23(10):3821-3831. doi: 10.1007/s00784-019-02811-5. Epub 2019 Jan 28.
In this study, we aimed to determine the suitable concentrations of human platelet lysate (HPL) and platelet-rich plasma (PRP) for maintaining the in vitro proliferative and angiogenic potential of inflamed dental pulp stem cells.
Lipopolysaccharide (LPS)-induced inflamed dental pulp-derived stem cells (iDPSCs) were treated with different concentrations of HPL and PRP (10% and 20%) followed by determination of viability using Alamar Blue assay. Expression of angiogenesis-, adhesion-, and inflammation-regulating genes was also analyzed using RT-qPCR array. Furthermore, expression of growth factors at protein level in the cell culture microenvironment was measured using multiplex assay.
Viability of iDPSCs was significantly (p < 0.05) higher in 20% HPL-supplemented media compared to iDPSCs. Expression of 10 out of 12 selected angiogenic genes, four out of seven adhesion molecules, and seven out of nine cytokine-producing genes were significantly (p < 0.05) higher in cells maintained in 20% HPL-supplemented media compared to that in FBS-supplemented media. Furthermore, expression of all the selected growth factors was significantly higher (p < 0.05) in the supernatants from 20% HPL media at 12 and 24 h post-incubation.
This study suggests that 20% HPL could be optimum to stimulate angiogenesis-related factors in iDPSCs while maintaining their viability.
This data may suggest the potential use of 20% HPL for expanding DPSCs scheduled for clinical trials for regenerative therapies including dental pulp regeneration.
本研究旨在确定人血小板裂解液（HPL）和富血小板血浆（PRP）的适宜浓度，以维持炎症牙髓干细胞的体外增殖和血管生成潜能。
用不同浓度的 HPL 和 PRP（10%和 20%）处理脂多糖（LPS）诱导的炎症牙髓来源的干细胞（iDPSCs），然后用 Alamar Blue 测定法测定细胞活力。使用 RT-qPCR 阵列分析血管生成、黏附和炎症调节基因的表达。此外，使用多重分析测量细胞培养微环境中生长因子的蛋白水平表达。
与 iDPSCs 相比，添加 20% HPL 的培养基中 iDPSCs 的活力显著更高（p<0.05）。与 FBS 补充培养基相比，在添加 20% HPL 的培养基中维持的细胞中，12 个选定的血管生成基因中的 10 个、7 个黏附分子中的 4 个和 9 个细胞因子生成基因中的 7 个的表达显著更高（p<0.05）。此外，在孵育 12 和 24 小时后，20% HPL 培养基的上清液中所有选定的生长因子的表达均显著更高（p<0.05）。
本研究表明，20% HPL 可能是刺激 iDPSCs 中与血管生成相关的因子的最佳选择，同时保持其活力。
该数据可能表明 20% HPL 可用于扩展用于再生疗法的 DPSCs，包括牙髓再生的临床试验。
