Procaine Inhibits Osteo/Odontogenesis through Wnt/β-Catenin Inactivation

Abstract

IntroductionPeriodontitis is a complex pathology characterized by the loss of alveolar bone. The causes and the mechanisms that promote this bone resorption still remain unknown. The knowledge of the critical regulators involved in the alteration of alveolar bone homeostasis is of great importance for developing molecular therapies. Procaine is an anesthetic drug with demethylant properties, mainly used by dentists in oral surgeries. The inhibitor role of Wnt signaling of procaine was described in vitro in colon cancer cells.MethodsIn this work we evaluated the role of procaine (1 uM) in osteo/odontogenesis of rat bone marrow mesenchymal stem cells. Similarly, the mechanisms whereby procaine achieves these effects were also studied.ResultsProcaine administration led to a drastic decrease of calcium content, alkaline phosphatase activity, alizarin red staining and an increase in the expression of Matrix Gla Protein. With respect to osteo/odontogenic markers, procaine decreased early and mature osteo/odontogenic markers. In parallel, procaine inhibited canonical Wnt/β-catenin pathway, observing a loss of nuclear β-catenin, a decrease in Lrp5 and Frizzled 3, a significant increase of sclerostin and Gsk3β and an increase of phosphorylated β-catenin. The combination of osteo/odontogenic stimuli and Lithium Chloride decreased mRNA expression of Gsk3β, recovered by Procaine. Furthermore it was proved that Procaine alone dose dependently increases the expression of Gsk3β and β-catenin phosphorylation. These effects of procaine were also observed on mature osteoblast. Interestingly, at this concentration of procaine no demethylant effects were observed.ConclusionsOur results demonstrated that procaine administration drastically reduced the mineralization and osteo/odontogenesis of bone marrow mesenchymal stem cells inhibiting Wnt/β-catenin pathway through the increase of Gsk3β expression and β-catenin phosphorylation.

Food for Bone: Evidence for a Role for Delta-Tocotrienol in the Physiological Control of Osteoblast Migration

“…As shown in Figure 7, pretreatment with procaine (2 µM) per se did not modify the MC3T3-E1 cell ability to cover the wound, but completely prevented the positive action of δ-TT on MC3T3-E1 cell migration. The involvement of β-catenin in the enhancing action of δ-TT on MC3T3-E1 cell migration was confirmed by the results obtained with procaine, an inhibitor of the Wnt/β-catenin signalling pathway [20,21]. As shown in Figure 7, pretreatment with procaine (2 μM) per se did not modify the MC3T3-E1 cell ability to cover the wound, but completely prevented the positive action of δ-TT on MC3T3-E1 cell migration.…”

“…Even if the lack of measurements of the intranuclear translocation of β-catenin represents a limitation of our study, we suggest that the canonical Wnt signalling pathway is involved in the effect of δ-TT on MC3T3-E1 cell migration since δ-TT treatment increased β-catenin transcriptional activity as detected by luciferase assay and pretreatment with procaine, a local anaesthetic drug, previously reported to inhibit the Wnt/β-catenin pathway [20,21], prevents the increase of the wound healing activity induced by δ-TT. Our assumption is in line with previous in vivo studies, showing that oral annatto TT supplementation prevented osteopenia induced by metabolic syndrome by reducing SOST and DKK1 levels, considered antagonist of the Wnt signalling pathway [33].…”

β-catenin and Its Relation to Alveolar Bone Mechanical Deformation – A Study Conducted in Rats With Tooth Extraction

“…It is necessary developing additional studies to verify if the β-catenin activation pattern corresponds spatially and temporally with bone remodeling against mechanical stimuli. In the literature, it has been consistent that β-catenin plays an important role in bone formation and remodeling (Herencia et al, 2016).…”

Effects of Nicotinamide on Cytotoxicity-induced Morphological Changes in Osteoblastic Cells <i>In Vitro</i>

“…This method is often used in studies on the proliferation and differentiation of osteoblasts [10][11][12][13][14] . In addition, they are used in studies involving bone metabolism and biomaterials [15][16][17][18][19] .…”