Interactive effects of LPS and dentine matrix proteins on human dental pulp stem cells
Interactive effects of LPS and dentine matrix proteins on human dental pulp stem cells
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DOI：
10.1111/iej.12897
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发表时间：
2018-08-01
影响因子：
5
通讯作者：
Galler, K. M.
中科院分区：
文献类型：
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作者：
Widbiller, M.;Eidt, A.;Galler, K. M.
目的探讨脂多糖（LPS）和提取的牙本质基质蛋白（eDMP）对人牙髓干细胞（DPSCs）再生和炎症反应的联合作用。方法在培养基中添加不同浓度的LPS、eDMP或两者的组合。MTT法测定1周内细胞活力；24h和7d后用流式细胞术测定细胞存活率。实时定量聚合酶链反应（RT-qPCR）检测矿化相关标记基因的表达。为了分析炎症反应，采用酶联免疫吸附法（ELISA）测定细胞培养初期和后期白细胞介素6 （IL-6）的分泌。资料采用非参数处理，各实验组间比较采用Mann-Whitney u检验（=0.05）。结果LPS对细胞活力无影响，eDMP对细胞活力呈浓度依赖性降低，7d后显著降低（P0.024）。48小时后，LPS诱导的细胞存活率中度下降（P0.026），而eDMP能够逆转这种影响。eDMP单独引起所测标记基因表达增加，LPS无调节作用。eDMP和LPS联合诱导I型胶原和骨钙素表达上调，而牙本质基质酸性磷蛋白和牙本质唾液磷蛋白的表达水平与对照组相似。随着时间的推移，LPS增加了il -6的分泌。eDMP显著提高初始IL-6的产生（P0.002），但抑制lps诱导的后期细胞因子的产生。结论聚多糖不影响成牙干细胞的细胞活力，但会干扰成牙干细胞样细胞的分化。来自牙本质基质的蛋白质可能具有保护作用，减轻LPS的有害影响，因此在牙髓修复中发挥重要作用。
AimTo investigate the combinatorial effects of lipopolysaccharide (LPS) and extracted dentine matrix proteins (eDMP) on regenerative and inflammatory responses in human dental pulp stem cells (DPSCs).MethodologyCulture media were supplemented with several concentrations of LPS, eDMP and combinations of both. Cell viability was assessed over 1week by MTT assay; cell survival was evaluated after 24h and 7days by flow cytometry. The expression of mineralization-associated marker genes was determined by real-time quantitative polymerase chain reaction (RT-qPCR). To analyse the inflammatory response, secretion of interleukin 6 (IL-6) was quantified in the initial and the late phase of cell culture by enzyme-linked immunosorbent assay (ELISA). Data were treated nonparametrically and Mann-Whitney U-tests were performed to compare all experimental groups (=0.05).ResultsWhereas LPS had no impact on viability, eDMP led to a concentration-dependent decrease, which was significant after 7days (P0.024). A moderate decline of cell survival induced by LPS was detected after 48h (P0.026), whereas eDMP was able to reverse this effect. eDMP alone caused increased expression of tested marker genes, LPS had no regulatory effect. Combined eDMP and LPS induced an upregulation of collagen type I and osteocalcin, whereas expression levels of dentine matrix acidic phosphoprotein and dentine sialophosphoprotein were similar to the control. IL-6-secretion was increased by LPS over time. eDMP markedly elevated initial production of IL-6 (P0.002), but suppressed LPS-induced cytokine production in the later phase.ConclusionsLipopolysaccharide did not affect cell viability but interfered with odontoblast-like cell differentiation of DPSCs. Proteins from the dentine matrix may have a protective effect, attenuate the detrimental impact of LPS and thus play an important role during pulp repair.