Abstract
Our study attempted to compare the efficacies of bone morphogenetic protein (BMP) 2, 6, and 9 in inducing osteogenic differentiation of preodontoblasts (PDBs). We immortalized PDBs by introducing a reversible SV40 T antigen-based immortalization system. Cell proliferation capability was examined by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay. The effects of BMP2, 6, and 9 on the osteogenic differentiation of immortalized preodontoblasts (iPDBs) were measured by alkaline phosphatase (ALP) activity assays and alizarin red S staining. The expression of osteogenic markers was evaluated by semiquantitative real-time polymerase chain reaction analysis. To assess ectopic bone formation, rat-derived iPDBs were transfected in culture with adenoviral vectors designated Ad-BMP2, 6, and 9 and subcutaneously or intramuscularly injected into mice. Several BMPs retained endogenous expression in PDBs and regulated the mRNA expression of mineralized tissue-associated proteins. ALP activity and mineralized nodule formation were significantly increased in the Ad-BMP9-transfected group relative to the control group. In addition, the most significant hard tissue formation was in this group. The results indicated that BMP signaling was involved in the osteogenic differentiation of iPDBs. BMP9 could be an efficacious accelerant of the osteogenic differentiation of iPDBs.
Highlights
Bone tissue regeneration is a current challenge in the treatment of hard tissue defects
To confirm that immortalized preodontoblasts (iPDBs) express bone morphogenetic protein (BMP)-related genes, we measured the endogenous expression of BMP2, 6, and 9 using an real-time polymerase chain reaction (RT-PCR) assay
bone sialoprotein (BSP), OPN, and OCN expression were significantly upregulated after induction with BMP2, 6, or 9 as early as day 3, while dentin matrix protein 1 (DMP1) and matrix extracellular phosphoglycoprotein (MEPE) expression were upregulated, achieving higher levels at later time points
Summary
Bone tissue regeneration is a current challenge in the treatment of hard tissue defects. The key components of bone tissue engineering are osteogenic growth factors, stem cells, and extracellular matrix scaffolds [1]. Our research group is developing a rational method based on morphogenetic signals for bone tissue induction, stem/ progenitor cell responses, and scaffolds to repair bone defects. Dental tissue-originated MSCs stand out as a stem cell source with broad potential for bone regeneration in various bone surgeries due to their convenient extraction, abundance, and easy isolation and purification [2]. Preodontoblasts (PDBs), the progenitors of odontoblasts, derive from dental papilla and have similar bone regeneration and restoration abilities, including cell proliferation and extracellular matrix maturation and mineralization [3]. We propose that exogenous bone morphogenetic proteins (BMPs) might have the capacity to induce bone tissue-associated gene mRNA expression and mineralization of human PDBs [4,5]
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Schedule a callMore From: Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas
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