Engineered scaffolds based on mesenchymal stem cells/preosteoclasts extracellular matrix promote bone regeneration

Abstract

Recently, extracellular matrix-based tissue-engineered bone is a promising approach to repairing bone defects, and the seed cells are mostly mesenchymal stem cells. However, bone remodelling is a complex biological process, in which osteoclasts perform bone resorption and osteoblasts dominate bone formation. The interaction and coupling of these two kinds of cells is the key to bone repair. Therefore, the extracellular matrix secreted by the mesenchymal stem cells alone cannot mimic a complex bone regeneration microenvironment, and the addition of extracellular matrix by preosteoclasts may contribute as an effective strategy for bone regeneration. Here, we established the mesenchymal stem cell/preosteoclast extracellular matrix -based tissue-engineered bones and demonstrated that engineered-scaffolds based on mesenchymal stem cell/ preosteoclast extracellular matrix significantly enhanced osteogenesis in a 3 mm rat femur defect model compared with mesenchymal stem cell alone. The bioactive proteins released from the mesenchymal stem cell/ preosteoclast extracellular matrix based tissue-engineered bones also promoted the migration, adhesion, and osteogenic differentiation of mesenchymal stem cells in vitro. As for the mechanisms, the iTRAQ-labeled mass spectrometry was performed, and 608 differentially expressed proteins were found, including the IGFBP5 and CXCL12. Through in vitro studies, we proved that CXCL12 and IGFBP5 proteins, mainly released from the preosteoclasts, contributed to mesenchymal stem cells migration and osteogenic differentiation, respectively. Overall, our research, for the first time, introduce pre-osteoclast into the tissue engineering of bone and optimize the strategy of constructing extracellular matrix–based tissue-engineered bone using different cells to simulate the natural bone regeneration environment, which provides new sight for bone tissue engineering.

Fabrication and Characterization of Collagen/PVA Dual-Layer Membranes for Periodontal Bone Regeneration

“…In addition to the collagen based artificial scaffolds or membranes, it is worthy to mention another promising type of scaffold, based more on the extracellular matrix as well as cells, named as “cell sheet” ( Chen et al, 2019 ; Lu et al, 2019 ). Cell sheet was efficaciously utilized in the regeneration of bone and periodontal ligament-like tissues as shown by recent works ( Dong et al, 2020 ; Liu Y. et al, 2020 ). Promising as it demonstrated to be, cell sheet manufacture requires high manipulation skills to ensure the quality control ( Chen et al, 2019 ), while the current artificial Col/PVA membrane offers the superiority in relatively easy manufacturing with controllability.…”

Clinical Application of Bone Marrow Mesenchymal Stem/Stromal Cells to Repair Skeletal Tissue

“…Alternative strategies have attempted to mimic the bone remodeling process, where bone anabolic osteoblast function and catabolic osteoclast function are reproduced. One report utilized mouse derived BMSC transplanted at a 10:1 ratio with mouse bone marrow mononuclear cells (BMMNC), containing pre-osteoclasts, combined with a decalcified bone matrix scaffold, which resulted in greater bone formation in a rat critical size defect model over BMSC transplanted alone or with no BMSC [ 125 ]. Supportive proteomic analysis and in vitro studies utilizing neutralizing antibodies suggest that the mechanism by which the combination of BMSC and pre-osteoclasts facilitates greater bone repair was mediated by the pre-osteoclast secretion of CXCL12, which promoted MSC migration; and Insulin Like Growth Factor Binding Protein 5 (IGFBP5), which enhanced MSC osteogenic differentiation [ 125 ].…”

“…One report utilized mouse derived BMSC transplanted at a 10:1 ratio with mouse bone marrow mononuclear cells (BMMNC), containing pre-osteoclasts, combined with a decalcified bone matrix scaffold, which resulted in greater bone formation in a rat critical size defect model over BMSC transplanted alone or with no BMSC [ 125 ]. Supportive proteomic analysis and in vitro studies utilizing neutralizing antibodies suggest that the mechanism by which the combination of BMSC and pre-osteoclasts facilitates greater bone repair was mediated by the pre-osteoclast secretion of CXCL12, which promoted MSC migration; and Insulin Like Growth Factor Binding Protein 5 (IGFBP5), which enhanced MSC osteogenic differentiation [ 125 ]. While others have also demonstrated CXCL12 stimulates MSC migration [ 95 , 110 ], CXCL12 is also abundantly expressed by MSC and osteogenic populations [ 126 , 127 ] and is known to attract monocytes and promote osteoclast development and function [ 128 , 129 ].…”

In vivo study of polyurethane and tannin-modified hydroxyapatite composites for calvarial regeneration

“…Elaborately designed artificial bone grafts should be biocompatible and biodegradable as well as osteoconductive, osteoinductive and angiogenesis promoting, providing temporary and gradually regressive substitutes in the bone defect sites, and actively promote bone regeneration in the degradation process. 2 , 5 , 9 – 18 However, the fabrication of ideal bone grafts are still challenging.…”