RESUMEN
The objective of this study was to evaluate the effects of oral fish cartilage hydrolysate (FCH) on symptoms and joint tissue structure in rat developing osteoarthritis induced surgically. Osteoarthritis was induced in the right knee of mature male Lewis rats (n = 12/group) by surgical transection of the anterior cruciate ligament (ACLT) combined with partial medial meniscectomy (pMMx). Two weeks after surgery, rats were treated orally with either control (sterile H2O) or FCH for four weeks. Pain and function were assessed by dynamic weight-bearing test (incapacitance test), electronic Von Frey (EVF; hindpaw allodynia threshold), and pressure algometer (knee allodynia threshold). Time and groups differences at each time point were evaluated using a mixed model. The histological features were evaluated eight weeks after surgery using OARSI score. Mann-Whitney test nonparametric test was applied to compare OARSI score. ACTL/pMMx surgery significantly reduced weight-bearing and increased allodynia and sensitivity thresholds of the operated paw/knee. Globally, FCH improved these parameters faster, but no significant difference between control and FCH groups was observed. Eight weeks after surgery, rats developed moderate OA lesions. Compared with control, FCH did not significantly modify OA lesion severity assessed using the OARSI score. In this mechanically induced OA model, 4 weeks of supplementation with FCH had no significant effect on cartilage lesion, but tends to accelerate pain relief and joint function recovery. This positive trend may have opened the way for further investigation of FCH as potential treatment of joint discomfort associated with OA.
RESUMEN
Objectives: Zeel T (Ze14) is a multicomponent medicinal product. Initial preclinical data suggested a preventive effect on cartilage degradation. Clinical observational studies demonstrated that Ze14 reduced symptoms of osteoarthritis (OA), including stiffness and pain. This study aimed to explore these effects further to better understand the mode of action of Ze14 on human OA chondrocytes in vitro. Methods: Primary chondrocytes were obtained from the knees of 19 OA patients and cultured either as monolayers or in alginate beads. 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. Protein quantification by ELISA was performed on selected genes from the culture medium and/or the cellular fractions of primary human OA chondrocyte cultures. 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. Ze14 significantly increased (mean 2.3-fold after 24 h, p = 0.0444 and 72 h, p = 0.0239) the CCN1 protein production in human OA chondrocytes. After 72 h, Ze14 significantly increased type II collagen pro-peptide production by mean 27% (p = 0.0147). For both time points CCN1 production by OA chondrocytes was correlated with aggrecan (r = 0.66, p = 0.0004) and type II collagen pro-peptide (r = 0.64, p = 0.0008) production. In alginate beads cultures, pro-MMP-13 was decreased by Ze14 from day 7-14 (from -16 to -25%, p < 0.05) and from day 17-21 (-22%, p = 0.0331) in comparison to controls. Conclusion: Ze14 significantly modified the expression of DUSP1, DEPP1, ZFP36/ZFP36L1, 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.