Tissue Eng Regen Med.  2018 Dec;15(6):781-791. 10.1007/s13770-018-0150-x.

Glucosamine Hydrochloride and N-Acetylglucosamine Influence the Response of Bovine Chondrocytes to TGF-β3 and IGF in Monolayer and Three-Dimensional Tissue Culture

• Pizzolatti ALA
• Gaudig F
• Seitz D
• Roesler CR
• Salmoria G

• KMID: 2441269
• DOI: http://doi.org/10.1007/s13770-018-0150-x

Abstract

BACKGROUND
Glucosamine hydrochloride (GlcN·HCl) has been shown to inhibit cell growth and matrix synthesis, but not with N-acetyl-glucosamine (GlcNAc) supplementation. This effect might be related to an inhibition of critical growth factors (GF), or to a different metabolization of the two glucosamine derivatives. The aim of the present study was to evaluate the synergy between GlcN·HCl, GlcNAc, and GF on proliferation and cartilage matrix synthesis. METHOD: Bovine chondrocytes were cultivated in monolayers for 48 h and in three-dimensional (3D) chitosan scaffolds for 30 days in perfusion bioreactors. Serum-free (SF) medium was supplemented with either growth factors (GF) TGF-β (5 ng mL₋₁) and IGF-I (10 ng mL₋₁), GlcN·HCl or GlcNAc at 1mM each or both. Six groups were compared according to medium supplementation: (a) SF control; (b) SF + GlcN·HCl; (c) SF + GlcNAc; (d) SF + GF; (e) SF + GF + GlcN·HCl; and (f) SF + GF + GlcNAc. Cell proliferation, proteoglycan, collagen I (COL1), and collagen II (COL2) synthesis were evaluated.
RESULTS
The two glucosamines showed opposite effects in monolayer culture: GlcN·HCl significantly reduced proliferation and GlcNAc significantly augmented cellular metabolism. In the 30 days 3D culture, the GlcN·HCl added to GF stimulated cell proliferation more than when compared to GF only, but the proteoglycan synthesis was smaller than GF. However, GlcNAc added to GF improved the cell proliferation and proteoglycan synthesis more than when compared to GF and GF/GlcN·HCl. The synthesis of COL1 and COL2 was observed in all groups containing GF.
CONCLUSION
GlcN·HCl and GlcNAc increased cell growth and stimulated COL2 synthesis in long-time 3D culture. However, only GlcNAc added to GF improved proteoglycan synthesis.

Keyword

Glucosamine hydrochloride; N-Acetyl-Glucosamine; Growth factors; Cartilage Engineering

MeSH Terms

Bioreactors
Cartilage
Cell Proliferation
Chitosan
Chondrocytes*
Collagen
Glucosamine*
Insulin-Like Growth Factor I
Intercellular Signaling Peptides and Proteins
Metabolism
Methods
Perfusion
Proteoglycans
Chitosan
Collagen
Glucosamine
Insulin-Like Growth Factor I
Intercellular Signaling Peptides and Proteins
Proteoglycans