Dentine matrix proteins: isolation and effects on human pulp cells

Abstract

A novel, effective and time-saving protocol for isolation and concentration of dentine matrix proteins is presented. As eDMP stimulates chemotaxis, differentiation and mineralization without affecting viability, endogenous dentine matrix proteins might be valuable for approaches to regenerate or engineer dental pulp.

Interactive effects of LPS and dentine matrix proteins on human dental pulp stem cells

“…Dentine matrix proteins were extracted from human teeth (eDMP) according to a protocol described before (Widbiller et al 2018). Therefore, dentine was pulverized (Mixer Mill MM 200, RETSCH GmbH, Haan, Germany) and resuspended in 10% EDTA (269 mmol L À1 ; EDTA Disodium Salt Dihydrate BioChemica; AppliChem GmbH, Darmstadt, Germany).…”

“…Therefore, previously established primer sequences (Widbiller et al 2018) were used for collagen type I a1 (COL1A1), dentine sialophosphoprotein (DSPP), dentine matrix acidic phosphoprotein 1 (DMP1), osteocalcin (OCN) and 40S ribosomal protein S18 (RPS18) as housekeeping gene (Table 1). Therefore, previously established primer sequences (Widbiller et al 2018) were used for collagen type I a1 (COL1A1), dentine sialophosphoprotein (DSPP), dentine matrix acidic phosphoprotein 1 (DMP1), osteocalcin (OCN) and 40S ribosomal protein S18 (RPS18) as housekeeping gene (Table 1).…”

“…LPS affected the viability of DPSCs (Liu et al 2014) and the proliferation of lung fibroblasts (Zhang et al 2011a) possibly through TLR-4-dependent pathways or cytokine release (IL-6 and IL-8). Antiproliferative effects of dentine-derived proteins have been described for various cell types (Zhang et al 2011b, Widbiller et al 2018, and TGF-b1, besides other molecules, may play an important role via the Smad-pathway (Massagu e 2000). In this study, the active cell metabolism (viability) was determined by a tetrazolium reduction assay (MTT assay) and an impact of LPS in the tested concentrations on human DPSCs during the incubation period of 1 week was not detected.…”

“…Amongst those are proteins from the SIBLING-family, such as dentine sialoprotein (DSP), dentine phosphoprotein (DPP), dentine matrix acidic phosphoprotein 1 (DMP1) and osteopontin (OPN). Once released by demineralization through carious acids or pulp capping agents (Dung et al 1995, Tomson et al 2007, these molecules stimulate migration of human pulp cells (Suzuki et al 2011, Galler et al 2016, Widbiller et al 2018) and induce odontoblastlike cell differentiation and mineralization (Tziafas et al 1995, Widbiller et al 2018. In addition, dentine contains angiogenic proteins such as vascular endothelial growth factor (VEGF) or platelet-derived growth factor (PDGF) as well as neurotrophins such as nerve growth factor (NGF) and glial cell-derived neurotrophic factor (GDNF) (Roberts-Clark & Smith 2000, Tomson et al 2017.…”

Isolation of primary odontoblasts: Expectations and limitations

“…After cleavage of the precursor dentin sialophosphoprotein (Dspp), the resulting dentin phosphoprotein (Dpp) and dentin sialoprotein (Dsp) play a role during mineralisation and are finally embedded in the dentin matrix. Thus, DSPP seems to be primarily expressed in odontoblasts or odontoblast-like cells during differentiation or progressive mineralisation (24,31). This might be due to the fact that most of the crown construction is finished after primary dentinogenesis and the odontoblasts resided in an inactive state.…”