Transcriptomics analysis revealed reduction of Matrix Metallopeptidase 13 and promotion of chondrogenesis by Zeel T in primary human osteoarthritic chondrocytes

Objectives: Zeel T (Ze14) is a multicomponent medicinal product. Initial preclinical data suggested a preventive effect on cartilage degradation. This study aimed to explore these effects further to better understand the mode of action of Ze14 on human osteoarthritic (OA) chondrocytes in vitro. Methods: Primary chondrocytes were obtained from the knees of 10 OA patients and cultured in monolayers. The cultures were treated with 20 % or 10 % (v/v) Ze14 or placebo. For RNA-Seq, reads were generated with Illumina NextSeq5000 sequencer and aligned to the human reference genome (UCSC hg19). Differential expression analysis between Ze14 and placebo was performed in R using the DESeq2 package. Results: In monolayer cultures, Ze14 20 % (v/v) significantly modified the expression of 13 genes in OA chondrocytes by at least 10 % with an adjusted p-value < 0.05 : EGR1, FOS, NR4A1, DUSP1, ZFP36, ZFP36L1, NFKBIZ and CCN1 were upregulated and ATF7IP, TXNIP, DEPP1, CLEC3A and MMP13 were downregulated after 24 h Ze14 treatment. Conclusion: Ze14 significantly modified the expression of DUSP1, DEPP1, ZFP36/L1 and CLEC3A, which may reduce MMP13 expression and activation. Protein analysis confirmed that Ze14 significantly reduced the production of pro-MMP-13. As MMP-13 is involved in type II collagen degradation, Ze14 may limit cartilage degradation. Ze14 also promoted extracellular matrix formation arguably through CCN1 production, a growth factor well correlated with type II collagen and aggrecan production. Overall design: RNA-Seq on primary human OA chondrocytes mRNA cultured in monolayer in the presence or not of 10-11M interleukin-1b and/or Zeel T (Ze14)