ABSTRACT
Bone regeneration strategies rely on biomaterial constructs with stem cells or growth factors. By comparison, cell homing strategies employ chemokines to recruit the host endogenous stem or progenitor cells to the defect site to support endogenous healing. In the present study, we used a novel fluffy hydroxyapatite/polyacrylonitrile (HA/PAN) composite scaffold to provide a better three-dimensional cell culture microenvironment. These HA/PAN composite scaffolds loaded with stromal cell-derived factor-1α (SDF-1α) provided a diffusion-controlled SDF-1α release profile and endowed the scaffolds with cell homing capabilities. Furthermore, the scaffolds significantly stimulated bone marrow stromal cell (BMSC) recruitment, facilitated BMSC osteogenic differentiation, and promoted ectopic bone formation. Our results suggest that a HA/PAN composite scaffold loaded with SDF-1α offers a clinically beneficial bone repair strategy.
ABSTRACT
Craniofacial defects can cause morbidness. Adipose-derived stem cells (ADSCs) have shown great promise for osteogeneration and vascularization; therefore cocultures of differentiated ADSCs are explored to increase bone and vessel formation. In this study, ADSCs were induced into osteogenic ADSCs (os-ADSCs) and endothelial ADSCs (endo-ADSCs) cells, which were then cocultured in variable proportions (os-ADSCs/endo-ADSCs = 2:1, 1:1, 1:2). The os-ADSCs in a ratio of 1:1 expressed more ALP, RUNX2 and COL-I, whereas VEGF, vWF and CD31 were upregulated in the endo-ADSCs of this group. Next generation RNA sequencing (RNA-seq) was performed to evaluate the molecular mechanisms of cocultured ADSCs. The os-ADSCs and endo-ADSCs interacted with each other during osteogenic and angiogenic differentiation, especially at the ratio of 1:1, and were regulated by vascular-related genes, cell-mediated genes, bone-related genes and the transforming growth factor ß signaling pathway (TGF-ß), mitogen-activated protein kinase signaling pathway (MAPK) and wnt signaling pathway (Wnt). Angptl4, apoe, mmp3, bmp6, mmp13 and fgf18 were detected to be up-regulated, and cxcl12 and wnt5a were down-regulated. The results showed that the gene expression levels were consistent with that in RNA-seq. The cells were then seeded into self-assembling peptide RADA16-I scaffolds as cocultures (1:1) and monocultures (ADSCs, os-ADSCs, endo-ADSCs). The results showed that the cells of all groups grew and proliferated well on the scaffolds, and the cocultured group exhibited better osteogeneration and vascularization. In conclusion, cocultured os-ADSCs and endo-ADSCs at the ratio of 1:1 showed strong osteogenic and angiogenic differentiation. There is a great potential for osteogenesis and vascularization by 3D culturing cells in a 1:1 ratio in self-assembling peptide RADA16-I scaffolds, which requires evaluation for bone regeneration in vivo.
Subject(s)
Adipose Tissue/cytology , Bone Regeneration/drug effects , Cell Differentiation/drug effects , Peptides/pharmacology , Stem Cells/cytology , Animals , Biomarkers/metabolism , Cell Differentiation/genetics , Cell Proliferation/drug effects , Coculture Techniques , Endothelial Cells/cytology , Endothelial Cells/drug effects , Endothelial Cells/metabolism , Gene Expression Regulation/drug effects , Neovascularization, Physiologic/drug effects , Osteogenesis/drug effects , Osteogenesis/genetics , Phenotype , Rats, Sprague-Dawley , Stem Cells/drug effects , Stem Cells/metabolism , Stem Cells/ultrastructure , Tissue Scaffolds/chemistryABSTRACT
In recent years, various composite scaffolds based on hydroxyapatite have been developed for bone tissue engineering. However, the poor cell survival micro-environment is still the major problem limiting their practical applications in bone repairing and regeneration. In this study, we fabricated a class of fluffy and porous three-dimensional composite fibrous scaffolds consisting of hydroxyapatite and polyacrylonitrile by employing an improved electrospinning technique combined with a bio-mineralization process. The fluffy structure of the hydroxyapatite/polyacrylonitrile composite scaffold ensured the cells would enter the interior of the scaffold and achieve a three-dimensional cell culture. Bone marrow mesenchymal stem cells were seeded into the scaffolds and cultured for 21 days in vitro to evaluate the response of cellular morphology and biochemical activities. The results indicated that the bone marrow mesenchymal stem cells showed higher degrees of growth, osteogenic differentiation and mineralization than those cultured on the two-dimensional hydroxyapatite/polyacrylonitrile composite membranes. The obtained results strongly supported the fact that the novel three-dimensional fluffy hydroxyapatite/polyacrylonitrile composite scaffold had potential application in the field of bone tissue engineering.