Promoted Chondrogenesis of Cocultured Chondrocytes and Mesenchymal Stem Cells under Hypoxia Using In-situ Forming Degradable Hydrogel Scaffolds

Biomacromolecules. 2018 Jan 8;19(1):94-102. doi: 10.1021/acs.biomac.7b01271. Epub 2017 Dec 18.

Abstract

We investigated the effects of different oxygen tension (21% and 2.5% O2) on the chondrogenesis of different cell systems cultured in pH-degradable PVA hydrogels, including human articular chondrocytes (hACs), human mesenchymal stem cells (hMSCs), and their cocultures with a hAC/hMSC ratio of 20/80. These hydrogels were prepared with vinyl ether acrylate-functionalized PVA (PVA-VEA) and thiolated PVA-VEA (PVA-VEA-SH) via Michael-type addition reaction. The rheology tests determined the gelation of the hydrogels was controlled within 2-7 min, dependent on the polymer concentrations. The different cell systems were cultured in the hydrogel scaffolds for 5 weeks, and the safranin O and GAG assay showed that hypoxia (2.5% O2) greatly promoted the cartilage matrix production with an order of hAC > hAC/hMSC > hMSC. The real time quantitative PCR (RT-PCR) revealed that the hMSC group exhibited the highest hypertrophic marker gene expression (COL10A1, ALPL, MMP13) as well as the dedifferentiated marker gene expression (COL1A1) under normoxia conditions (21% O2), while these expressions were greatly inhibited by coculturing with a 20% amount of hACs and significantly further repressed under hypoxia conditions, which was comparative to the sole hAC group. The enzyme-linked immunosorbent assay (ELISA) also showed that coculture of hMSC/hAC greatly reduced the catabolic gene expression of MMP1 and MMP3 compared with the hMSC group. It is obvious that the hypoxia conditions promoted the chondrogenesis of hMSC by adding a small amount of hACs, and also effectively inhibited their hypotrophy. We are convinced that coculture of hAC/hMSC using in situ forming hydrogel scaffolds is a promising approach to producing cell source for cartilage engineering without the huge needs of primary chondrocyte harvest and expansion.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkaline Phosphatase / metabolism
  • Biocompatible Materials / chemistry
  • Cartilage, Articular / cytology
  • Cartilage, Articular / enzymology
  • Cartilage, Articular / metabolism
  • Cell Hypoxia*
  • Chondrocytes / cytology*
  • Chondrocytes / enzymology
  • Chondrocytes / metabolism
  • Chondrogenesis*
  • Coculture Techniques
  • Collagen / genetics
  • Enzyme-Linked Immunosorbent Assay
  • Gene Expression
  • Glycosaminoglycans / metabolism
  • Humans
  • Hydrogels / chemistry*
  • Matrix Metalloproteinase 13 / genetics
  • Mesenchymal Stem Cells / cytology*
  • Mesenchymal Stem Cells / enzymology
  • Mesenchymal Stem Cells / metabolism
  • Middle Aged
  • Phenazines / metabolism
  • Polyvinyl Alcohol / chemistry
  • Tissue Scaffolds*

Substances

  • Biocompatible Materials
  • Glycosaminoglycans
  • Hydrogels
  • Phenazines
  • Polyvinyl Alcohol
  • Collagen
  • Alkaline Phosphatase
  • MMP13 protein, human
  • Matrix Metalloproteinase 13
  • safranine T