Sun Wei, Ma Dandan, Bolscher Jan G M, Nazmi Kamran, Veerman Enno C I, Bikker Floris J, Sun Ping, Lin Haiyan, Wu Gang
The Affiliated Stomatology Hospital, Zhejiang University School of Medicine, Hangzhou, China.
Key Laboratory of Oral Biomedical Research of Zhejiang Province, Hangzhou, China.
Front Bioeng Biotechnol. 2020 Oct 23;8:584410. doi: 10.3389/fbioe.2020.584410. eCollection 2020.
Promoting cell spreading is crucial to enhance bone healing and implant osteointegration. In this study, we investigated the stimulatory effect of human salivary histatin-1 (Hst-1) on the spreading of osteogenic cells as well as the potential signaling pathways involved. Osteogenic cells were seeded on bio-inert glass slides with or without the presence of Hst1 in dose-dependent or time-course assays. 1 scrambled and 6 truncated Hst1 variants were also evaluated. Cell spreading was analyzed using a well-established point-counting method. Fluorescent microscopy was adopted to examine the cellular uptake of fluorescently labeled Hst1 (F-Hst1) and also the cell spreading on sandblasted and acid etched titanium surfaces. Signaling inhibitors, such as U0126, SB203580, and pertussis toxin (PTx) were used to identify the potential role of extracellular-signal-regulated kinase, p38 and G protein-coupled receptor pathways, respectively. After 60 min incubation, Hst1 significantly promoted the spreading of osteogenic cells with an optimal concentration of 10 μM, while truncated and scrambled Hst1 did not. F-Hst1 was taken up and localized in the vicinity of the nuclei. U0126 and SB2030580, but not PTx, inhibited the effect of Hst1. 10 μM Hst1 significantly promoted the spreading of osteogenic cells on both bio-inert substrates and titanium SLA surfaces, which involved ERK and p38 signaling. Human salivary histatin-1 might be a promising peptide to enhance bone healing and implant osteointegration in clinic.
促进细胞铺展对于增强骨愈合和植入物骨整合至关重要。在本研究中，我们研究了人唾液组蛋白-1（Hst-1）对成骨细胞铺展的刺激作用以及相关的潜在信号通路。在存在或不存在Hst1的情况下，将成骨细胞接种在生物惰性载玻片上，进行剂量依赖性或时间进程分析。还评估了1种乱序和6种截短的Hst1变体。使用成熟的点计数法分析细胞铺展。采用荧光显微镜检查荧光标记的Hst1（F-Hst1）的细胞摄取情况以及在喷砂和酸蚀钛表面上的细胞铺展情况。分别使用信号抑制剂，如U0126、SB203580和百日咳毒素（PTx）来确定细胞外信号调节激酶、p38和G蛋白偶联受体途径的潜在作用。孵育60分钟后，Hst1以10μM的最佳浓度显著促进成骨细胞铺展，而截短和乱序的Hst1则无此作用。F-Hst1被摄取并定位在细胞核附近。U0126和SB2030580而非PTx抑制了Hst1的作用。10μM Hst1显著促进了成骨细胞在生物惰性底物和钛SLA表面上的铺展，这涉及ERK和p38信号传导。人唾液组蛋白-1可能是临床上增强骨愈合和植入物骨整合的一种有前景的肽。
