CGRP gene-modified rBMSCs show better osteogenic differentiation capacity in vitro

Abstract

Calcitonin gene-related peptide (CGRP) is a marked and important neuropeptide expressed in nerve fibers during bone repair. This study investigated the role of CGRP overexpression on osteogenic differentiation of rat bone mesenchymal stem cells (rBMSCs). rBMSCs were infected with viral stocks of pLenO-DCE-CGRP (CGRP group) or pLenO-DCE (Vector group), while normal rBMSCs were used as a control. Transfection efficiency of rBMSCs was analyzed by flow cytometry. Cell proliferation was examined using a Cell Counting Kit-8 and flow cytometry. Expressions of alkaline phosphatase(ALP), bone sialoprotein (BSP) and Runt-related transcription factor 2(Runx2) in rBMSCs were detected at 1 and 2 weeks after mineral induction by real-time PCR and western blotting. Alizarin Red staining was applied at 28 days. The ratio of osteoprotegerin (OPG) to receptor activator of nuclear factor kappa B ligand (RANKL) was also detected to determine the underlying mechanism. pLenO-DCE-CGRP-induced rBMSCs stably overexpressing CGRP were successfully established. Overexpression of the CGRP gene significantly promoted rBMSC proliferation (p < 0.05). In addition, expressions of osteogenesis-related indexes were upregulated in the CGRP group (p < 0.05) compared with vector and control groups, and more mineralization nodules were observed in the CGRP group (p < 0.05). CGRP gene increased OPG and reduced RANKL in rBMSCs. Hence, the OPG/ RANKL ratio was increased in the CGRP group compared with the other two groups. CGRP gene-modified rBMSCs show better osteogenic differentiation capacity compared with rBMSCs in vitro.

miRNA-21 promotes osteogenesis via the PTEN/PI3K/Akt/HIF-1α pathway and enhances bone regeneration in critical size defects

“…Genetically modified stem cells have been shown to play significant roles in promoting bone regeneration [5]. Previous studies have reported that a variety of factors, such as transcription factors, growth factors, cytokines, and extracellular matrix molecules, can promote osteogenesis and angiogenesis in bone tissue engineering [6].…”

PLGA Containing Human Adipose-Derived Stem Cell-Derived Extracellular Vesicles Accelerates the Repair of Alveolar Bone Defects via Transfer of CGRP

“…Analysis on the interaction network of candidate genes by the GeneMANIA tool indicated that six genes had higher interaction scores, including DMP1 (score = 0.867194116), SOST (score = 0.814230232), SP7 (score = 0.786778992), IBSP (score = 0.758063414), MEN1 (score = 0.737518121), and CALCA (score = 0.733867971) ( Figure 2(b) ). Existing evidence has confirmed the correlation between CGRP and osteogenic differentiation of hPDLSCs [ 19 , 20 ]. CGRP was also predicted to be upregulated in the GSE53929 dataset ( Figure 2(c) ).…”

Calcitonin Gene-Related Peptide Influences Bone-Tendon Interface Healing Through Osteogenesis: Investigation in a Rabbit Partial Patellectomy Model

“…4 In vitro, CGRP may regulate bone metabolism by inhibiting osteoclast formation, 8 whereas studies have shown that CGRP can promote bone mesenchymal stem cell osteogenic differentiation and osteogenic ability linked to the higher expression of osteogenesis-related indexes alkaline phosphatase, bone sialoprotein, and runt-related transcription factor 2. 13,35 Recombinant human CGRP can induce rabbit adipose-derived stem cells into osteoblast differentiation. 11 This study has several limitations.…”