Sulfated hyaluronan/collagen I matrices enhance the osteogenic differentiation of human mesenchymal stromal cells in vitro even in the absence of dexamethasone

Acta Biomater. 2012 Nov;8(11):4064-72. doi: 10.1016/j.actbio.2012.06.039. Epub 2012 Jul 6.


Glycosaminoglycans (GAG) are multifunctional components of the extracellular matrix (ECM) involved in different steps of the regulation of cellular differentiation. In this study artificial extracellular matrices (aECM) consisting of collagen (Col) I and different GAG derivatives were used as a substrate for human mesenchymal stromal cells (hMSC) to study osteogenic differentiation in vitro. hMSC were cultured on aECM containing col and hyaluronan sulfates (HyaS) with increasing degrees of sulfation (DS(S)) and were compared with aECM containing col and the natural GAG hyaluronan or chondroitin 4-sulfate. hMSC were analyzed for osteogenic differentiation markers such as calcium phosphate deposition, tissue non-specific alkaline phosphatase (TNAP) and expression of runt-related transcription factor 2 (runx2), osteocalcin (ocn) and bone sialoprotein II (bspII). Compared with aECM containing Col and natural GAG all Col/HyaS-containing aECM induced an increase in calcium phosphate deposition, TNAP activity and tnap expression. These effects were also seen in the absence of dexamethasone (an established osteogenic supplement). The expression of runx2 and ocn was not altered and the expression of bspII was diminished on the col/HyaS-containing aECM. The impact of the Col/HyaS-containing aECM on hMSC differentiation was independent of the DS(S) of the HyaS derivatives, indicating the importance of the primary (C-6) hydroxyl group of N-acetylglucosamine. These results suggest that Col/HyaS-containing aECM are able to stimulate hMSC to undergo osteogenic differentiation even in the absence of dexamethasone, which makes these matrices an interesting tool for hMSC-based tissue engineering applications and biomaterial functionalizations to enhance bone formation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Alkaline Phosphatase / genetics
  • Alkaline Phosphatase / metabolism
  • Animals
  • Calcium Phosphates / metabolism
  • Cell Differentiation / drug effects*
  • Collagen Type I / pharmacology*
  • Core Binding Factor Alpha 1 Subunit / genetics
  • Core Binding Factor Alpha 1 Subunit / metabolism
  • Dexamethasone / pharmacology*
  • Extracellular Matrix / drug effects
  • Extracellular Matrix / metabolism
  • Gene Expression Regulation / drug effects
  • Glycosaminoglycans / chemistry
  • Glycosaminoglycans / metabolism
  • Humans
  • Hyaluronic Acid / chemical synthesis
  • Hyaluronic Acid / chemistry
  • Hyaluronic Acid / pharmacology*
  • Integrin-Binding Sialoprotein / genetics
  • Integrin-Binding Sialoprotein / metabolism
  • Male
  • Mesenchymal Stem Cells / cytology*
  • Mesenchymal Stem Cells / drug effects
  • Mesenchymal Stem Cells / enzymology
  • Osteocalcin / genetics
  • Osteocalcin / metabolism
  • Osteogenesis / drug effects
  • Rats
  • Staining and Labeling
  • Sulfates / pharmacology*


  • Calcium Phosphates
  • Collagen Type I
  • Core Binding Factor Alpha 1 Subunit
  • Glycosaminoglycans
  • Integrin-Binding Sialoprotein
  • Sulfates
  • Osteocalcin
  • Dexamethasone
  • Hyaluronic Acid
  • calcium phosphate
  • Alkaline Phosphatase