Enhancement of chondrogenesis of human adipose derived stem cells in a hyaluronan-enriched microenvironment.
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.
Wu
Shun-Cheng
SC
Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
Chang
Je-Ken
JK
Wang
Chih-Kuang
CK
Wang
Gwo-Jaw
GJ
Ho
Mei-Ling
ML
D017667:Adipocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D053674:Aggrecans / Q000235:genetics; D001672:Biocompatible Materials / Q000737:chemistry / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D020219:Chondrogenesis / Q000187:drug effects*; D024042:Collagen Type I / Q000235:genetics; D024043:Collagen Type II / Q000235:genetics; D024121:Collagen Type X / Q000235:genetics; D006025:Glycosaminoglycans / Q000378:metabolism; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry*; D007150:Immunohistochemistry; D019344:Lactic Acid / Q000737:chemistry; D011100:Polyglycolic Acid / Q000737:chemistry; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D016133:Polymerase Chain Reaction; D055758:SOX9 Transcription Factor / Q000235:genetics; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry
D053674:Aggrecans; D001672:Biocompatible Materials; D024042:Collagen Type I; D024043:Collagen Type II; D024121:Collagen Type X; D006025:Glycosaminoglycans; D055758:SOX9 Transcription Factor; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; C013786:A73025; D019344:Lactic Acid; D006820:Hyaluronic Acid