Carvacrol induces osteogenic differentiation of BMSCs and alleviates osteolysis in aged mice by upregulating lncRNA NEAT1 to promote SIRT1 expression

Abstract

Background: Wear particle-induced periprosthetic osteolysis is a major contributor to joint replacement failure and revision surgery in the elderly. Osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) is a promising approach for bone regeneration. Carvacrol may have bone-protective potential. This study investigated whether carvacrol alleviates osteolysis and promotes osteogenic differentiation of BMSCs by regulating SIRT1 expression.

Materials and Methods: An osteolysis model in aged mice was established by titanium (Ti) particle induction. BMSCs were isolated from femur and tibia of 18-month-old mice and cultured in osteogenic differentiation medium. RIP and RNA pull-down were used to evaluate the binding of SIRT1 and lncRNA NEAT1. Ubiquitination analysis was performed to investigate NEAT1-mediated regulation of SIRT1 ubiquitination modification levels.

Results: Carvacrol treatment improved Ti particle-induced calvarial erosion and attenuated osteolysis. Carvacrol increased SIRT1 both in the mouse model of Ti-induced osteolysis and in BMSCs under osteogenic differentiation. Carvacrol treatment promoted ALP activity, bone mineralization capacity, and expression of osteogenic differentiation-related factors, whereas SIRT1 knockdown reversed this effect. LncRNA NEAT1 interacted with SIRT1, and overexpression of NEAT1 stabilized SIRT1 by inhibiting its ubiquitination modification. NEAT1 knockdown altered the promoting effect of carvacrol on osteogenic differentiation of BMSCs, while overexpression of SIRT1 reversed the effects of NEAT1 knockdown. Similarly, NEAT1 knockdown altered the inhibitory effect of carvacrol on osteolysis in vivo.

Conclusion: Our research results demonstrate that carvacrol showed potential in treating osteolysis in aged mice by regulating NEAT1 to promote the expression of SIRT1 and can facilitate the osteogenic differentiation of BMSCs.

Keywords: Osteolysis, carvacrol, bone marrow-derived mesenchymal stem cells, NEAT1, SIRT1