Evaluation of the effects of pregabalin on chondrocyte proliferation and CHAD, HIF-1 α, and COL2A1 gene expression

Abstract

Introduction: The aim of the present study is to investigate the effects of pregabalin (PGB) on chondrocyte proliferation and collagen type II (COL2A1), hypoxia-inducible factor 1-α (HIF-1α), and chondroadherin (CHAD) gene expression in osteoarthritic chondrocytes.

Material and methods: Standard primary chondrocyte cultures were prepared using osteochondral tissues that were surgically obtained from 6 patients with gonarthrosis. Cell morphology was evaluated using an inverted microscope, and cell death and proliferation were determined through MTT analysis, which was confirmed by AO/PI staining and statistically evaluated. The expression levels of CHAD, COL2A1, and HIF-1α genes were assessed using gene-specific TaqMan Gene Expression Assays.

Results: MTT analyses showed that PGB administration did not have a negative or toxic effect on cell viability and proliferation in cultured chondrocytes (p < 0.001), but in our morphological evaluation extracellular matrix development was observed to be weaker in cultures treated with PGB. After 24 h of treatment, COL2A1, HIF-1α, and CHAD gene expression decreased in the groups to which PGB was applied compared to gene expression before the experiment (at 0 h); at 48 h, CHAD and HIF-1α expression increased to the same level as the control group, but the expression of COL2A1 continued to decrease.

Conclusions: Further studies need to be conducted with more participants to prove that there is a negative correlation between extracellular matrix formation and PGB administration. Our preliminary data show that even at low doses and over short-term administration, PGB may affect chondrocyte cells at the gene-expression level.

Keywords: CHAD; COL2A1; HIF-1α; osteoarthritic chondrocytes; pregabalin.

Figure 1

Morphological evaluation of chondrocyte cultures Group 1 – control samples at 0, 24 and 48 h. Group 2 – PGB-treated group at 0, 24, and 48 h.

Figure 2

AO/PI staining of chondrocyte cultures Group 1 – control samples at 0, 24, and 48 h. Group 2 – PGB-treated group at 24 and 48 h.

Figure 3

Relative quantification values obtained from experimental groups Group 1 – control samples 0, 24, and 48 h. Group 2 – PGBtreated group at 24 and 48 h.