Carbon monoxide releasing molecule‑3 promotes the osteogenic differentiation of rat bone marrow mesenchymal stem cells by releasing carbon monoxide

Abstract

Stem cell‑based therapies are promising strategies to stimulate bone regeneration. Carbon monoxide releasing molecule‑3 (CORM‑3) exhibits multiple regulatory effects in a number of cells by releasing carbon monoxide (CO). The present study aimed to investigate the influence of CORM‑3 on the osteogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs). BMSCs were divided into five groups: A CORM‑3‑osteogenic group, in which cells were pretreated with CORM‑3 and subjected to osteogenic differentiation induction using osteogenic medium; an osteogenic group, in which cells were cultured in osteogenic medium; a degassed CORM‑3‑osteogenic group, in which cells were pretreated with degassed CORM‑3 and subjected to osteogenic differentiation induction; a CORM‑3 group, in which cells were cultured in control medium containing CORM‑3; and a control group, in which cells were cultured in control medium alone. The osteo‑specific mRNA and protein expression of runt‑related transcription factor 2 (Runx2), osteocalcin (OCN) and osteopontin (OPN) were assessed using reverse transcription‑quantitative polymerase chain reaction and western blot analysis. Alkaline phosphatase (ALP) activity was also examined and mineralization was detected using alizarin red staining. Levels of Runx2, OCN and OPN mRNA and protein in the CORM‑3‑osteogenic group were significantly increased compared with the osteogenic group (P<0.05), with the exception of OCN protein levels on day 3. The mRNA and protein expression of Runx2, OCN and OPN in the degassed CORM‑3‑osteogenic and osteogenic groups were similar. In addition, the mRNA and protein expression of Runx2, OCN and OPN in the CORM‑3 and control group were similar. ALP activity in the CORM‑3‑osteogenic group was increased from day 3 and remained significantly higher compared with all other groups on days 3, 5 and 7 (P<0.05). Additionally, the results indicated that the optical density value of alizarin red staining in the CORM‑3‑osteogenic group was significantly increased compared with the other groups (P<0.05). Therefore, the present study demonstrated that CORM‑3 may promote the osteogenic differentiation of BMSCs by releasing CO.

Extracellular vesicle-encapsulated miR-22-3p from bone marrow mesenchymal stem cell promotes osteogenic differentiation via FTO inhibition

“…RUNX2 belongs to the RUNX family and plays an important role in osteoblast differentiation and skeletal formation [23]. Moreover, RUNX serves as a biomarker for early stage of osteogenic differentiation while OCN and OPN are both osteoblast markers [24]. Therefore, induction of osteogenic differentiation would be accompanied by upregulation of RUNX1, OCN, and OPN.…”

Carbon Monoxide and Exercise Prevents Diet‐Induced Obesity and Metabolic Dysregulation Without Affecting Bone

“…The disparity between OP+EX and OP+COEX rats is not easily attributed to hormonal responses because leptin, adiponectin, and IGF‐1 levels were similar. CO does suppress osteoclastogenesis and stimulates osteoblastogenesis , yet because obesity alone did not result in bone loss, this is questionable. Instead and more plausibly, the further reduction in body mass caused by CO plus aerobic exercise was responsible for the differences in femoral material properties between OP+EX and OP+COEX rats.…”

“…In parallel in vitro experiments using mouse bone marrow cells, the authors reported that CORM‐2 inhibited osteoclastogenesis by reducing receptor activator of nuclear factor‐κB ligand (RANKL) and oxidant production . In addition, CO was reported to increase proliferation of bone marrow mesenchymal stem cells and their differentiation into osteoblasts versus adipocytes . Indeed, diet‐induced obesity was shown to not only induce osteoclastogenesis, in part through RANKL activation, but also to increase bone marrow adipocyte expansion in mice and rats .…”

Carbon Monoxide Releasing Molecule 3 Inhibits Osteoclastogenic Differentiation of RAW264.7 Cells by Heme Oxygenase-1

“…There is an abundance of preclinical evidence in both large and small animal models of diseases demonstrating the protective effects of CO at low concentrations (15-250 ppm) and CORMs, including hyperacute endotoxic shock [26], pulmonary inflammation [27], postoperative ileus [28], organ transplantation-induced ischemia-reperfusion injury [29,[30][31][32][33], airway hyperresponsiveness [34], necrotizing enterocolitis [35], and pulmonary hypertension [36]. CORM-3 has also been reported to promote the osteogenic differentiation of bone marrow mesenchymal stem cells by releasing CO [37].…”