Fibrin-Hyaluronic Acid Hydrogel (RegenoGel) with Fibroblast Growth Factor-18 for In Vitro 3D Culture of Human and Bovine Nucleus Pulposus Cells

Int J Mol Sci. 2019 Oct 11;20(20):5036. doi: 10.3390/ijms20205036.

Abstract

We investigated the effects of a fibrin-hyaluronic acid hydrogel (FBG-HA) and fibroblast growth factor 18 (FGF-18) for nucleus pulposus (NP) regeneration. Healthy bovine (n = 4) and human degenerated NP cells (n = 4) were cultured for 14 days in FBG-HA hydrogel with FGF-18 (∆51-mutant or wild-type) in the culture medium. Gene expression, DNA content, and glycosaminoglycan (GAG) synthesis were evaluated on day 7 and 14. Additionally, histology was performed. Human NP cells cultured in FBG-HA hydrogel showed an increase in collagen type II (COL2) and carbonic anhydrase XII (CA12) gene expression after 14 or 7 days of culture, respectively. GAG release into the conditioned medium increased over 14 days. Healthy bovine NP cells showed increased gene expression of ACAN from day 7 to day 14. Wild type FGF-18 up-regulated CA12 gene expression of human NP cells. Histology revealed an increase of proteoglycan deposition upon FGF-18 stimulation in bovine but not in human NP cells. The FBG-HA hydrogel had a positive modulatory effect on human degenerated NP cells. Under the tested conditions, no significant effect of FGF-18 was observed on cell proliferation or GAG synthesis in human NP cells.

Keywords: fibrin-hyaluronic acid hydrogel; fibroblast growth factor 18; nucleus pulposus cells.

MeSH terms

  • Animals
  • Biomimetics
  • Carbonic Anhydrases / genetics
  • Cattle
  • Cell Culture Techniques / methods*
  • Cells, Cultured
  • Collagen Type II / metabolism
  • Fibroblast Growth Factors / chemistry
  • Fibroblast Growth Factors / pharmacology*
  • Glycosaminoglycans / metabolism
  • Humans
  • Hyaluronic Acid / chemistry*
  • Hyaluronic Acid / pharmacology
  • Hydrogels / chemistry
  • Nucleus Pulposus / cytology*
  • Nucleus Pulposus / drug effects
  • Nucleus Pulposus / metabolism
  • Phenotype
  • Regeneration

Substances

  • Collagen Type II
  • Glycosaminoglycans
  • Hydrogels
  • fibroblast growth factor 18
  • Fibroblast Growth Factors
  • Hyaluronic Acid
  • Carbonic Anhydrases
  • carbonic anhydrase XII