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        Glucosamine Hydrochloride and N-Acetylglucosamine Influence the Response of Bovine Chondrocytes to TGF-b3 and IGF in Monolayer and Three-Dimensional Tissue Culture

        Andre´ Luiz A. Pizzolatti,Florian Gaudig,Daniel Seitz,Carlos R. M. Roesler,Gean Vitor Salmoria 한국조직공학과 재생의학회 2018 조직공학과 재생의학 Vol.15 No.6

        BACKGROUND: Glucosamine hydrochloride (GlcNHCl) 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 GlcNHCl, 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-b (5 ng mL-1) and IGF-I (10 ng mL-1), GlcNHCl or GlcNAc at 1mM each or both. Six groups were compared according to medium supplementation: (a) SF control; (b) SF ? GlcNHCl; (c) SF ? GlcNAc; (d) SF ? GF; (e) SF ? GF ? GlcNHCl; 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: GlcNHCl significantly reduced proliferation and GlcNAc significantly augmented cellular metabolism. In the 30 days 3D culture, the GlcNHCl 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/GlcNHCl. The synthesis of COL1 and COL2 was observed in all groups containing GF. CONCLUSION: GlcNHCl and GlcNAc increased cell growth and stimulated COL2 synthesis in long-time 3D culture. However, only GlcNAc added to GF improved proteoglycan synthesis.

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