Human Salivary Histatin-1 Promotes Osteogenic Cell Spreading on Both Bio-Inert Substrates and Titanium SLA Surfaces

Abstract

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 in vitro 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.

Human Salivary Histatin-1-Functionalized Gelatin Methacrylate Hydrogels Promote the Regeneration of Cartilage and Subchondral Bone in Temporomandibular Joints

“…Therefore, it may also be explained that Hst1 induced the secretion of growth factors and cytokines from mature chondrocytes so as to stimulate the chondrogenic differentiation of hematopoietic stem cells. It has recently been established that Hst1 promotes osteogenic differentiation [ 57 , 58 ]. Furthermore, our recent study shows that Hst1 significantly promotes BMP2-induced angiogenesis and osteogenesis [ 31 ].…”

“…Therefore, the interaction of exogenous Hst1 and endogenous BMP2 may be also responsible for the promotion of cartilage formation in the current study. Previous studies suggested that Hst’s effect is associated with numerous signaling pathways, such as RAC1 [ 30 ], ERK [ 23 ], p38 [ 57 ], and NF-ĸB [ 58 ], among others. However, there has been no report on the effect of Hst1 on chondrocytes.…”

The Impact of Early Saliva Interaction on Dental Implants and Biomaterials for Oral Regeneration: An Overview

“…The authors demonstrated that saliva interaction with biomaterials prior to cell culture impaired the behavior of the MC3T3-E1 osteoblast cell line [14], MG63 human osteoblasts [52] and bone marrow cells derived from Sprague-Dawley rats [21] (Figure 3); additionally, more reports showed the same problematic behavior for soft tissue cells such as human gingival fibroblasts (HGFs) [47,53]. On the other hand, Sun et al [54] revealed that the addition of an isolated salivary protein (histatin-1) to a titanium surface improved the spread and some features of MC3T3-E1 osteoblast cells after culture for some days. Moreover, Caballe-Serrano et al [55] demonstrated, using in vitro models, that bone tissue contaminated with saliva showed less osteoclast reabsorption and presented some differences in the immune response, which highlights the need for further investigation regarding the effects of saliva interaction in clinical or animal models.…”

“…Table 2 summarizes studies involving different cells and their reactions to contaminated substrates with saliva, suggesting in 100% of the studies that saliva impairs the behavior or proliferation of specific cells. The opposite is only revealed when studies propose the isolation of specific proteins (histatin-1) to adhere to the substrate and create a beneficial effect for the cells [54,60]. Zhou et al [47] In vitro HGFs cell seemed over the surfaces.…”

Using a two-step method of surface mechanical attrition treatment and calcium ion implantation to promote the osteogenic activity of mesenchymal stem cells as well as biomineralization on a β-titanium surface

“…14 Hence, surface modication provides a potential avenue to enhance the biological performance, simultaneously preserving the outstanding bulk properties of titanium-based implants. 15 There are various routes by which to modify titanium-based metals by monitoring surface roughness, topography or grain structure to achieve good biological properties, such as etching, 16,17 sandblasting, 18 shot peening 19 and sliding friction treatment (SFT). 20 Among them, nanograined titanium-based surfaces with microroughness generated by the surface mechanical attrition treatment (SMAT) approach have been utilized effectively to benet cell response in vitro [21][22][23][24] and enhance bone formation capacity in vivo.…”