Enzyme-crosslinked gene-activated matrix for the induction of mesenchymal stem cells in osteochondral tissue regeneration
Journal
Acta Biomaterialia
Journal Volume
63
Pages
210-226
Date Issued
2017
Author(s)
Lee, Yi-Hsuan
Wu, Hsi-Chin
Yeh, Chia-Wei
Liao, Han-Tsung
Hsu, Horng-Chaung
Tsai, Jui-Che
Abstract
The development of osteochondral tissue engineering is an important issue for the treatment of traumatic injury or aging associated joint disease. However, the different compositions and mechanical properties of cartilage and subchondral bone show the complexity of this tissue interface, making it challenging for the design and fabrication of osteochondral graft substitute. In this study, a bilayer scaffold is developed to promote the regeneration of osteochondral tissue within a single integrated construct. It has the capacity to serve as a gene delivery platform to promote transfection of human mesenchymal stem cells (hMSCs) and the functional osteochondral tissues formation. For the subchondral bone layer, the bone matrix with organic (type I collagen, Col) and inorganic (hydroxyapatite, Hap) composite scaffold has been developed through mineralization of hydroxyapatite nanocrystals oriented growth on collagen fibrils. We also prepare multi-shell nanoparticles in different layers with a calcium phosphate core and DNA/calcium phosphate shells conjugated with polyethyleneimine to act as non-viral vectors for delivery of plasmid DNA encoding BMP2 and TGF-β3, respectively. Microbial transglutaminase is used as a cross-linking agent to crosslink the bilayer scaffold. The ability of this scaffold to act as a gene-activated matrix is demonstrated with successful transfection efficiency. The results show that the sustained release of plasmids from gene-activated matrix can promote prolonged transgene expression and stimulate hMSCs differentiation into osteogenic and chondrogenic lineages by spatial and temporal control within the bilayer composite scaffold. This improved delivery method may enhance the functionalized composite graft to accelerate healing process for osteochondral tissue regeneration. Statement of Significance In this study, a gene-activated matrix (GAM) to promote the growth of both cartilage and subchondral bone within a single integrated construct is developed. It has the capacity to promote transfection of human mesenchymal stem cells (hMSCs) and the functional osteochondral tissues formation. The results show that the sustained release of plasmids including TGF-beta and BMP-2 from GAM could promote prolonged transgene expression and stimulate hMSCs differentiation into the osteogenic and chondrogenic lineages by spatial control manner. This improved delivery method should enhance the functionalized composite graft to accelerate healing process in vitro and in vivo for osteochondral tissue regeneration. ? 2017 Acta Materialia Inc.
SDGs
Other Subjects
Biomechanics; Bone; Calcium phosphate; Cartilage; Collagen; Crosslinking; Gene expression; Gene transfer; Histology; Hydroxyapatite; Molecular biology; Scaffolds (biology); Stem cells; Tissue regeneration; Bi-layer; Cross linking; Enzymatic cross-linking; Gene-activated matrix; Human mesenchymal stem cells; Mesenchymal stem cell; Osteochondral; Osteochondral graft substitute; Osteochondral tissues; Tissue formation; Cell culture; alkaline phosphatase; bone morphogenetic protein 2; calcium phosphate; collagen type 1; hydroxyapatite; nanocomposite; osteopontin; plasmid DNA; polyethyleneimine; protein glutamine gamma glutamyltransferase; transcription factor RUNX2; transforming growth factor beta3; bone morphogenetic protein 2; cross linking reagent; DNA; transforming growth factor beta3; Article; articular cartilage; bone development; bone mineralization; cell culture; cell differentiation; cell viability; chondrogenesis; collagen fiber; collagen fibril; compression; controlled study; differential scanning calorimetry; elasticity; energy dispersive X ray spectroscopy; extracellular matrix; gene activation; HeLa cell line; human; human cell; human tissue; hyaline cartilage; immunohistochemistry; joint; lipid bilayer; mesenchymal stem cell; priority journal; protein cross linking; protein expression; real time polymerase chain reaction; sustained drug release; tissue engineering; tissue regeneration; bone development; chemistry; chondrogenesis; cytology; gene; gene expression regulation; genetic transfection; infrared spectroscopy; mesenchymal stroma cell; metabolism; photon correlation spectroscopy; plasmid; regeneration; tissue scaffold; ultrastructure; Bone Morphogenetic Protein 2; Calorimetry, Differential Scanning; Chondrogenesis; Collagen Type I; Cross-Linking Reagents; DNA; Durapatite; Dynamic Light Scattering; Extracellular Matrix; Gene Expression Regulation; Genes; HeLa Cells; Humans; Mesenchymal Stromal Cells; Osteogenesis; Plasmids; Polyethyleneimine; Regeneration; Spectroscopy, Fourier Transform Infrared; Tissue Scaffolds; Transfection; Transforming Growth Factor beta3; Transglutaminases
Publisher
Acta Materialia Inc
Type
journal article