A novel decellularized matrix of Wnt signaling-activated osteocytes accelerates the repair of critical-sized parietal bone defects with osteoclastogenesis, angiogenesis, and neurogenesis

• Published in: Bioactive materials 2023-03, Vol.21, p.110-128
• Main Authors: Wang, Xiaofang, Ma, Yufei, Chen, Jie, Liu, Yujiao, Liu, Guangliang, Wang, Pengtao, Wang, Bo, Taketo, Makoto M., Bellido, Teresita, Tu, Xiaolin
• Format: Article
• Language: English
• Subjects: 3D printing; Decellularized matrix; Metabolic and neurovascular organoid bone; Osteocyte; Regenerative repair; Wnt signaling
• ISSN: 2452-199X; 2452-199X
• DOI: 10.1016/j.bioactmat.2022.07.017

Cell source is the key to decellularized matrix (DM) strategy. This study compared 3 cell types, osteocytes with/without dominant active Wnt/β-catenin signaling (daCO and WTO) and bone marrow stromal cells (BMSCs) for their DMs in bone repair. Decellularization removes all organelles and >95% DNA, and retained >74% collagen and >71% GAG, maintains the integrity of cell basement membrane with dense boundaries showing oval and honeycomb structure in osteocytic DM and smooth but irregular shape in the BMSC-DM. DM produced higher cell survival rate (90%) and higher proliferative activity. In vitro, daCO-DM induces more and longer stress fibers in BMSCs, conducive to cell adhesion, spreading, and osteogenic differentiation. 8-wk after implantation of the critical-sized parietal bone defect model, daCO-DM formed tight structures, composed of a large number of densely-arranged type-I collagen under polarized light microscope, which is similar to and integrated with host bone. BV/TV (>54%) was 1.5, 2.9, and 3.5 times of WTO-DM, BMSC-DM, and none-DM groups, and N.Ob/T.Ar (3.2 × 102/mm2) was 1.7, 2.9, and 3.3 times. At 4-wk, daCO-DM induced osteoclastogenesis, 2.3 times higher than WTO-DM; but BMSC-DM or none-DM didn't. daCO-DM increased the expression of RANKL and MCSF, Vegfa and Angpt1, and Ngf in BMSCs, which contributes to osteoclastogenesis, angiogenesis, and neurogenesis, respectively. daCO-DM promoted H-type vessel formation and nerve markers β3-tubulin and NeuN expression. Conclusion: daCO-DM produces metabolic and neurovascularized organoid bone to accelerate the repair of bone defects. These features are expected to achieve the effect of autologous bone transplantation, suitable for transformation application. [Display omitted] •Decellularized matrix of osteocytes with dominant-active β-catenin (daCO-DM) promotes osteogenesis for regenerative repair.•daCO-DM induces BMSCs to form stress fibers, conducive to cell adhesion, spreading, and differentiation towards osteoblasts.•daCO-DM-induced osteoblasts have strong activity secreting dense and orderly-arranged type I collagen as host bone’s.•daCO-DM induces BMSCs to express pre-osteoclastogenic cytokine RANKL and MCSF for osteoclastogenesis of marrow monocytes.•daCO-DM enhances BMSCs to express angiogenic Vegfa and Angpt1, and neurogenic Ngf potentially for neurovascularization.