Human saliva exposure modulates bone cell performance in vitro

Abstract

Various situations encountered by a clinician during the daily routine including surgical periodontitis therapy, dental implant insertion, or tooth extraction involve the contact of saliva with the jaw bone. However, there are only sparse data concerning the influence of saliva on bone cells. Saliva specimens were incorporated within culture medium and administered to murine MC3T3 osteoblasts, of which the morphology (REM), proliferation (EZ4U), and differentiation (qRT-PCR, alkaline phosphatase activity, extracellular matrix calcification) were assessed. Simultaneously, the composition of saliva media was analyzed with respect to the content of lactoferrin, activities of classical salivary enzymes, and the ability to provoke inflammatory cytokine production (enzyme-linked immunosorbent assay) in MC3T3 osteoblasts. The morphology, proliferation, and expression of differentiation-associated genes were seriously handicapped by saliva contact. Saliva-touched cells exhibited less alkaline phosphatase but normal levels of extracellular matrix mineralization. Saliva-containing culture media featured physiological activities of salivary enzymes and considerable amounts of lactoferrin but almost completely lacked salivary alkaline phosphatase and unspecific proteases. Upon saliva incubation, MC3T3 osteoblasts did not release noteworthy levels of interleukin-1 beta or tumor necrosis factor alpha. Although saliva is generally considered to vitalize oral tissues, this study reveals that it harms osteoblast-like cells more due to the presence of salivary enzymes than by triggering of inflammation. This issue is clinically relevant because it broadens the understanding of the bone cell fate within the rather complex cosmos of the oral cavity thereby providing a basis for clinical decision making and treatment guidelines. It seems to be reasonable to restrict the contact period between saliva and bone.

Differential inflammatory response of dental pulp explants and fibroblasts to saliva

“…For example, human saliva modulates proliferation and differentiation of osteogenic MC3T3-E1 cells in vitro (Proksch et al 2012). Parotid saliva enhanced IL-8 production of human leukaemia cells (Proksch et al 2012). Also previous data implicate a strong inflammatory response of gingiva fibroblasts and periodontal ligament fibroblasts, as indicated by an increase in IL-6 and IL-8 expression (Cvikl et al 2014).…”

“…For example, human saliva modulates proliferation and differentiation of osteogenic MC3T3‐E1 cells in vitro (Proksch et al . ). Parotid saliva enhanced IL‐8 production of human leukaemia cells (Proksch et al .…”

“…Parotid saliva enhanced IL‐8 production of human leukaemia cells (Proksch et al . ). Also previous data implicate a strong inflammatory response of gingiva fibroblasts and periodontal ligament fibroblasts, as indicated by an increase in IL‐6 and IL‐8 expression (Cvikl et al .…”

“…MC3T3‐E1 cells, however, failed to release notable amounts of IL‐1 or tumour necrosis factor‐alpha (TNF‐α) in response to saliva (Proksch et al . ). Thus, the impact of saliva on the inflammatory response of cells remains unclear, and the reaction of dental pulp fibroblasts to saliva has yet to be determined.…”

“…The small number of studies on the cellular impact of saliva reflects this ambiguity. For example, human saliva modulates proliferation and differentiation of osteogenic MC3T3-E1 cells in vitro (Proksch et al 2012). Parotid saliva enhanced IL-8 production of human leukaemia cells (Proksch et al 2012).…”