Chondrogenic differentiation of ChM-I gene transfected rat bone marrow-derived mesenchymal stem cells on 3-dimensional poly (L-lactic acid) scaffold for cartilage engineering

Cell Biol Int. 2015 Mar;39(3):300-9. doi: 10.1002/cbin.10393. Epub 2014 Dec 17.

Abstract

We have explored the role of Chondromodulin-I (ChM-I) in chondrogenesis of bone marrow-derived mesenchymal stem cells (BMSCs) in 3-dimensional (3D) scaffold for cartilage tissue engineering. BMSCs of Sprague Dawley (SD) rats were cultured on poly-(L-lactic acid) [PLLA] scaffolds with different pore sizes (80-200 μm, 200-450 μm) with or without surface modification by chitosan. Cell viability, proliferation, and morphology were measured using confocal microscope and the CCK-8 method. Untransfected BMSCs, BMSCs expressing pcDNA3.1(+), BMSCs expressing plasmid pcDNA3.1 (+)/ChM-I were cultured on 3D scaffolds in standard growth medium or transforming growth factor-β1 (TGF-β1) supplemented chondrogenic induction medium in vitro for 3 weeks and the expression of collagen type II was determined. Cell-scaffolds constructs were implanted subcutaneously for 3 months in vivo. BMSCs had a higher viability and proliferation in PLLA scaffolds of pore size 200-450 μm than that of 80-200 μm, and surface modification with chitosan did not enhance cell attachment. The ChM-I gene enhanced chondrogenesis and increased collagen type II synthesis. Immunohistochemistry from in vivo study showed enhanced cartilage regeneration in BMSCs expressing pcDNA3.1 (+)/ChM-I on 3D PLLA scaffolds. It also demonstrated that TGF-β1 might promote chondrogenesis of rat BMSCs by synergizing with the ChM-I gene. ChM-I could be beneficial to future applications in cartilage repair.

Keywords: ChM-I gene; cartilage tissue engineering; poly- (L-lactic acid); rat BMSCs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bone Marrow Cells / cytology*
  • Cartilage / metabolism
  • Cartilage / physiology
  • Cell Culture Techniques
  • Cell Differentiation / drug effects
  • Cell Proliferation
  • Cells, Cultured
  • Chondrogenesis / drug effects
  • Collagen Type II / metabolism
  • Intercellular Signaling Peptides and Proteins / genetics
  • Intercellular Signaling Peptides and Proteins / metabolism*
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Mesenchymal Stem Cell Transplantation
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / metabolism*
  • Polylysine / chemistry*
  • Rats
  • Rats, Sprague-Dawley
  • Regeneration / drug effects
  • Tissue Engineering*
  • Tissue Scaffolds
  • Transfection
  • Transforming Growth Factor beta1 / pharmacology

Substances

  • Cnmd protein, rat
  • Collagen Type II
  • Intercellular Signaling Peptides and Proteins
  • Membrane Proteins
  • Transforming Growth Factor beta1
  • Polylysine