The synergistic effect of micro-topography and biochemical culture environment to promote angiogenesis and osteogenic differentiation of human mesenchymal stem cells.
Critical failures associated with current engineered bone grafts involve insufficient induction of osteogenesis of the implanted cells and lack of vascular integration between graft scaffold and host tissue. This study investigated the combined effects of surface microtextures and biochemical supplements to achieve osteogenic differentiation of human mesenchymal stem cells (hMSCs) and revascularization of the implants in vivo. Cells were cultured on 10μm micropost-textured polydimethylsiloxane (PDMS) substrates in either proliferative basal medium (BM) or osteogenic medium (OM). In vitro data revealed that cells on microtextured substrates in OM had dense coverage of extracellular matrix, whereas cells in BM displayed more cell spreading and branching. Cells on microtextured substrates in OM demonstrated a higher gene expression of osteoblast-specific markers, namely collagen I, alkaline phosphatase, bone sialoprotein, and osteocalcin, accompanied by substantial amount of bone matrix formation and mineralization. To further investigate the osteogenic capacity, hMSCs on microtextured substrates under different biochemical stimuli were implanted into subcutaneous pockets on the dorsal aspect of immunocompromised mice to study capacity for ectopic bone formation. In vivo data revealed greater expression of osteoblast-specific markers coupled with increased vascular invasion on microtextured substrates with hMSCs cultured in OM. Together, these data represent a novel regenerative strategy that incorporates defined surface microtextures and biochemical stimuli to direct combined osteogenesis and re-vascularization of engineered bone scaffolds for musculoskeletal repair and relevant bone tissue engineering applications.
Department of Bioengineering and Therapeutic Sciences, University of California - San Francisco, San Francisco, CA 94158, United States.
Shang
S
Song
Department of Bioengineering and Therapeutic Sciences, University of California - San Francisco, San Francisco, CA 94158, United States.
Eun Jung
EJ
Kim
Department of Orthopaedic Surgery, University of California, San Francisco, Orthopaedic Trauma Institute, University of California, San Francisco/San Francisco General Hospital, San Francisco, CA 94110, United States.
Chelsea S
CS
Bahney
Department of Orthopaedic Surgery, University of California, San Francisco, Orthopaedic Trauma Institute, University of California, San Francisco/San Francisco General Hospital, San Francisco, CA 94110, United States.
Theodore
T
Miclau
Department of Orthopaedic Surgery, University of California, San Francisco, Orthopaedic Trauma Institute, University of California, San Francisco/San Francisco General Hospital, San Francisco, CA 94110, United States.
Ralph
R
Marcucio
Department of Bioengineering and Therapeutic Sciences, University of California - San Francisco, San Francisco, CA 94158, United States. Electronic address: shuvo.roy@ucsf.edu.
Shuvo
S
Roy
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D002452:Cell Count; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media / Q000494:pharmacology; D004129:Dimethylpolysiloxanes / Q000494:pharmacology; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000648:ultrastructure; D051379:Mice; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D018919:Neovascularization, Physiologic / Q000187:drug effects / Q000235:genetics; D010012:Osteogenesis / Q000187:drug effects / Q000235:genetics; D019919:Prosthesis Implantation; D054457:Tissue Scaffolds / Q000737:chemistry
D003470:Culture Media; D004129:Dimethylpolysiloxanes; D016326:Extracellular Matrix Proteins; C013830:baysilon; D000469:Alkaline Phosphatase
Human mesenchymal stem cells (hMSCs); Microelectromechanical systems (MEMS); Microfabrication; Polydimethylsiloxane (PDMS); Surface topography