Enhancement of chondrogenesis of human adipose derived stem cells in a hyaluronan-enriched microenvironment

Biomaterials. 2010 Feb;31(4):631-40. doi: 10.1016/j.biomaterials.2009.09.089. Epub 2009 Oct 12.

Abstract

Microenvironment plays a critical role in guiding stem cell differentiation. We investigated the enhancing effect of a hyaluronan (HA)-enriched microenvironment on human adipose derived stem cell (hADSC) chondrogenesis for articular cartilage tissue engineering. The hADSCs were obtained from patients undergoing hip replacement. HA-coated wells and HA-modified poly-(lactic-co-glycolic acid) (HA/PLGA) scaffolds were used as the HA-enriched microenvironment. The mRNA expressions of chondrogenic (SOX-9, aggrecan and collagen type II), fibrocartilage (collagen type I), and hypertrophic (collagen type X) marker genes were quantified by real-time polymerase chain reaction. Sulfated glycosaminoglycan (sGAG) deposition was detected by Alcian blue, safranin-O staining, and dimethylmethylene blue (DMMB) assays. Localized collagen type II was detected by immunohistochemistry. The hADSCs cultured in HA-coated wells (0.005-0.5 mg/cm(2)) showed enhanced aggregation and mRNA expressions (SOX-9, collagen type II, and aggrecan) after 24h, and sGAG content was also significantly increased after 9 days of culture. The HA-modified PLGA did not change the cell adherence and viability of hADSCs. The mRNA expressions of chondrogenic marker genes were significantly enhanced in hADSCs cultured in HA/PLGA rather than those cultured in the PLGA scaffold after 1, 3, and 5 days of culture. The hADSCs cultured in HA/PLGA produced higher levels of sGAG and collagen type II, compared to those in the PLGA scaffold after 4 weeks of cultures. Our results suggest that HA-enriched microenvironment induces chondrogenesis in hADSCs, which may be beneficial in articular cartilage tissue engineering.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adipocytes / cytology*
  • Adipocytes / drug effects
  • Adipocytes / metabolism
  • Aggrecans / genetics
  • Biocompatible Materials / chemistry
  • Biocompatible Materials / pharmacology*
  • Cell Differentiation / drug effects
  • Chondrogenesis / drug effects*
  • Collagen Type I / genetics
  • Collagen Type II / genetics
  • Collagen Type X / genetics
  • Glycosaminoglycans / metabolism
  • Humans
  • Hyaluronic Acid / chemistry*
  • Immunohistochemistry
  • Lactic Acid / chemistry
  • Polyglycolic Acid / chemistry
  • Polylactic Acid-Polyglycolic Acid Copolymer
  • Polymerase Chain Reaction
  • SOX9 Transcription Factor / genetics
  • Stem Cells / cytology*
  • Stem Cells / drug effects
  • Stem Cells / metabolism
  • Tissue Engineering / methods*
  • Tissue Scaffolds / chemistry

Substances

  • Aggrecans
  • Biocompatible Materials
  • Collagen Type I
  • Collagen Type II
  • Collagen Type X
  • Glycosaminoglycans
  • SOX9 Transcription Factor
  • Polylactic Acid-Polyglycolic Acid Copolymer
  • Polyglycolic Acid
  • A73025
  • Lactic Acid
  • Hyaluronic Acid