Abstract
BackgroundMesenchymal stem cell (MSC)-based tissue engineering plays a major role in regenerative medicine. However, the efficiency of MSC transplantation and survival of engrafted stem cells remain challenging. Melatonin can regulate MSC biology. However, its function in the osteogenic differentiation of dental pulp-derived MSCs (DPSCs) remains unclear. We investigated the effects and mechanisms of melatonin on the osteogenic differentiation and bone regeneration capacities of DPSCs.MethodsThe biological effects and signaling mechanisms of melatonin with different concentrations on DPSCs were evaluated using a proliferation assay, the quantitative alkaline phosphatase (ALP) activity, Alizarin red staining, a real-time polymerase chain reaction, and a western blot in vitro cell culture model. The in vivo bone regeneration capacities were assessed among empty control, MBCP, MBCP + DPSCs, and MBCP + DPSCs + melatonin preconditioning in four-created calvarial bone defects by using micro-computed tomographic, histological, histomorphometric, and immunohistochemical analyses after 4 and 8 weeks of healing.ResultsIn vitro experiments revealed that melatonin (1, 10, and 100 μM) significantly and concentration-dependently promoted proliferation, surface marker expression (CD 146), ALP activity and extracellular calcium deposition, and osteogenic gene expression of DPSCs (p < 0.05). Melatonin activated the protein expression of ALP, OCN, and RUNX-2 and inhibited COX-2/NF-κB expression. Furthermore, the phosphorylation of mitogen-activated protein kinase (MAPK) p38/ERK signaling was significantly increased in DPSCs treated with 100 μM melatonin, and their inhibitors significantly decreased osteogenic differentiation. In vivo experiments demonstrated that bone defects implanted with MBCP bone-grafting materials and melatonin-preconditioned DPSCs exhibited significantly greater bone volume fraction, trabecular bone structural modeling, new bone formation, and osteogenesis-related protein expression than the other three groups at 4 and 8 weeks postoperatively (p < 0.05).ConclusionsThese results suggest that melatonin promotes the proliferation and osteogenic differentiation of DPSCs by regulating COX-2/NF-κB and p38/ERK MAPK signaling pathways. Preconditioning DPSCs with melatonin before transplantation can efficiently enhance MSCs function and regenerative capacities.
Highlights
Mesenchymal stem cells (MSCs) have self-renewal and multilineage differentiation capacities, including osteoblasts, chondrocytes, and adipocytes [1]
Chan et al Stem Cell Research & Therapy (2022) 13:73. These results suggest that melatonin promotes the proliferation and osteogenic differentiation of Dental pulp-derived MSCs (DPSCs) by regulating COX-2/NF-κB and p38/ERK mitogen-activated protein kinase (MAPK) signaling pathways
After the dental pulp tissues were washed with phosphate-buffered saline (PBS) three times and minced into pieces, smaller pieces of pulp tissue were cultured in 3.5-cm-diameter Petri dishes with growth medium (GM) containing α-minimum essential medium (α-MEM; Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 15% fetal bovine serum, with 1000 U/mL penicillin, 1000 μg/mL streptomycin, and 0.25 μg/mL amphotericin B (Thermo Fisher Scientific, Waltham, MA, USA) in an incubator at 37 °C with 5% CO2
Summary
Mesenchymal stem cells (MSCs) have self-renewal and multilineage differentiation capacities, including osteoblasts, chondrocytes, and adipocytes [1]. They promote tissue regeneration during bone repair in osteoporosis, bone fracture, osteoarthritis, and bone defects and are considered a good cell source in regenerative medicine [2, 3]. Mesenchymal stem cell (MSC)-based tissue engineering plays a major role in regenerative medicine. Its function in the osteogenic differentiation of dental pulp-derived MSCs (DPSCs) remains unclear.
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
