In vitro chondrogenesis of mesenchymal stem cells in recombinant silk-elastinlike hydrogels

Pharm Res. 2008 Mar;25(3):692-9. doi: 10.1007/s11095-007-9282-8. Epub 2007 Apr 3.

Abstract

Purpose: In this study the chondrocytic differentiation and cartilage matrix accumulation of human mesenchymal stem cells (hMSCs) were investigated after encapsulation in a genetically engineered silk-elastinlike protein polymer SELP-47 K as an injectable matrix for delivery of cell-based therapeutics.

Materials and methods: hMSCs were encapsulated in SELP-47 K and cultured for 4 weeks in chondrogenic medium with or without transforming growth factor-beta3 (TGF). Chondrogenic differentiation was evaluated by histological, RNA and biochemical analyses for the expression of cartilage extracellular matrix components.

Results: Histological and immunohistochemical staining revealed that the cells acquired a rounded morphology and were embedded in significant amounts of chondrogenic extracellular matrix. Reverse transcriptase (RT)-PCR showed an up-regulation in aggrecan, type II and type X collagen and SOX9 in presence of TGF-beta3. By day 28, constructs cultured in the presence of TGF-beta3 exhibited significant increase in sulfated glycosaminoglycan and total collagen content up to 65 and 300%, respectively.

Conclusions: This study demonstrates that SELP-47 K hydrogel can be used as a scaffold for encapsulation and chondrogenesis of hMSCs. The ability to use recombinant techniques to precisely control SELP structure enables the investigation of injectable protein polymer scaffolds for soft-tissue engineering with varied physicochemical properties.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Aggrecans / metabolism
  • Cartilage / metabolism
  • Cell Culture Techniques
  • Cell Differentiation
  • Cell Shape
  • Cell Survival
  • Cells, Cultured
  • Chondrocytes / metabolism*
  • Chondrogenesis* / genetics
  • Collagen Type II / metabolism
  • Collagen Type X / metabolism
  • Elastin / chemistry
  • Elastin / metabolism*
  • Elastin / toxicity
  • Glycosaminoglycans / metabolism
  • High Mobility Group Proteins / metabolism
  • Humans
  • Hydrogels*
  • Insect Proteins / chemistry
  • Insect Proteins / metabolism*
  • Insect Proteins / toxicity
  • Mesenchymal Stem Cells / metabolism*
  • RNA, Messenger / metabolism
  • Recombinant Proteins / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • SOX9 Transcription Factor
  • Silk / chemistry
  • Silk / metabolism*
  • Silk / toxicity
  • Time Factors
  • Tissue Scaffolds*
  • Transcription Factors / metabolism
  • Transforming Growth Factor beta3 / metabolism
  • Up-Regulation

Substances

  • Aggrecans
  • Collagen Type II
  • Collagen Type X
  • Glycosaminoglycans
  • High Mobility Group Proteins
  • Hydrogels
  • Insect Proteins
  • RNA, Messenger
  • Recombinant Proteins
  • SOX9 Transcription Factor
  • SOX9 protein, human
  • Silk
  • Transcription Factors
  • Transforming Growth Factor beta3
  • Elastin