3D-Printed β-Tricalcium Phosphate Scaffolds Promote Osteogenic Differentiation of Bone Marrow-Deprived Mesenchymal Stem Cells in an N6-methyladenosineDependent Manner

Abstract

Bone defect is a serious orthopedic disease which has been studied for a long time. Alternative degradable biomaterials are required for bone repairing and regeneration to address the limitation of autogenous bone. β-tricalcium phosphate (β-TCP) is an alternative material with good cytocompatibility and has been used in bone defect treatment. However, whether β-TCP contributes to osteogenesis of bone marrow stem cells (BMSCs) through N6-methyladenosine (m6A) modification remains unknown. To address this issue, we verified the effects of β-TCP on osteogenesis of BMSCs. We also studied the expression of m6A-related enzymes in BMSCs after β-TCP treatment. Furthermore, the m6A level and stability of Runt-related transcription factor 2 (RUNX2) mRNA were investigated after β-TCP treatment. Finally, rat calvarial defect models were performed to detect expression level of osteogenic factors and m6A-related enzymes after the stimulation of three-dimension (3D)-printed β-TCP scaffolds. We found that β-TCP showed good biocompatibility and was osteoinductive. Meanwhile, methyltransferase-like 3 (METTL3) increased, causing the elevation of m6A level of RUNX2, results in stabler RUNX2 mRNA level. At last, based on the animal experiments, we demonstrated that the increase of RUNX2 and METTL3 levels was induced by β-TCP. These findings suggest that METTL3 increases the m6A level of RUNX2 mRNA after β-TCP induction, contributing to its stability, and the results in vivo also confirmed the osteogenic and bone-repair properties of β-TCP.

The Dual Angiogenesis Effects via Nrf2/HO-1 Signaling Pathway of Melatonin Nanocomposite Scaffold on Promoting Diabetic Bone Defect Repair

“…The promising results from various clinical trials underscore the efficacy of β-TCP in bone repair, demonstrating outcomes comparable to allografts. 52 Our results showed that compared with the PCL scaffold, the PCL/β-TCP scaffold can effectively promote osteogenic differentiation of MC3T3 cells ( Figure 3 ). Moreover, our in vivo and in vitro results showed that the PCL/β-TCP/SA/MT group exhibited the best osteogenic ability ( Figures 3 and 7 ) compared to the other groups under diabetic conditions, which aligns with the findings of prior investigations.…”

Immediate to short-term inflammatory response to biomaterial implanted in calvarium of mice

“…33 However, Jiao et al reported that the use of stem cells along with the printed scaffolds composed of beta-tricalcium phosphate could help the stability of the enzymes and biological mechanisms effective in repair and bone formation, and its use is suggested for bone repair. 34 Despite the results of this study, the identification of surgical treatments with high effectiveness in healing large bone defects remains a challenge. The 3D printed scaffolds composed of betatricalcium phosphate, which are associated with unique biological and molecular properties, can be effective in increasing effectiveness of surgical treatment, but larger studies are still needed.…”

N6-Methyladenosine in Cell-Fate Determination of BMSCs: From Mechanism to Applications

“…The increased osteogenesis of BMSCs by m 6 A modification could also be used in the treatment of bone defects combined with degradable biomaterials. Compared to the control group, BMSCs stimulated by β-tricalcium phosphate (β-TCP) exhibited considerably greater expression of METTL3, which influences m 6 A modification of RNA in BMSCs and improves the stability of RUNX2 mRNA [ 151 ]. In animal bone defect models, Jiao et al.…”

“…In animal bone defect models, Jiao et al. [ 151 ] discovered that β-TCP increased the m 6 A alteration of RUNX2, which stimulated the production of new bone. Han and colleagues [ 152 ] revealed similar results by using mesenchymal stem cells of the apical papillas (SCAPs), odontogenic MSCs with strong osteo/odontogenic capacity.…”