28 October 2015
OleoGrapeSEED - powerful antioxidant
A study shows that OleoGrapeSEED, extract from olive and grape seed, may be of clinical significance for the treatment of osteoarthritis
By Elsa Mevel, Grap’Sud PhD Student
Osteoarthritis (OA) is a major health concern that affects a growing part of the aging population and is associated with strong socio-economic burdens [1, 2]. The optimal treatment would consist in a therapy that can arrest the progressive degradation and the loss of articular cartilage and improve relief of symptoms such as inflammation and pain. However, so far there is no curative treatment for OA.
It is well known that the inflammatory cytokines such as IL-1ß play important roles in OA [3, 4]. IL-1ß is thus well known to stimulate the synthesis of nitric oxide (NO), prostaglandin E2 (PGE2) and MMP-13 [5, 6]. Procyanidins (PCy) and hydroxytyrosol (HT), major bioactive polyphenols present in grape seed and olive, exert numerous health promoting effects counteracting inflammation, aging and cancer. In the light of these promising data we were interested in deciphering whether a combination of PCy and HT, using OleoGrape®SEED, an extract from olive and grape seed, could have chondroprotective actions and, in a clinical perspective, conserve the bioactivity after oral consumption.
Human chondrocytes and cartilage explants were harvested from tibial plateau and femoral condyles of human cadavers to test whether PCy/HT using OleoGrape®SEED can counteract IL-1ß effects. In these cellular models, the cytotoxicity was evaluated using a Methyl tetrazolium salt (MTS) assay. The products released (NO, PGE2 and MMP-13) were measured by the Griess method and by Enzyme-linked immunosorbent assay (ELISA), respectively. Then, the anti-IL-1ß effects of PCy/HT metabolites found in sera after oral consumption were evaluated. For the in vivo study, rabbits (6 per groups) underwent a destabilisation of the right joint induced by anterior cruciate ligament transection (ACLT). OleoGrape®SEED or vehicle (Veh) were administrated every two days by stomacal gavages for 13 weeks, starting 3 weeks before surgery. Knee OA severity was quantitatively assessed by X-ray blind evaluation using a homemade radiographic score performed by 2 independent readers.
Results indicated that OleoGrape® SEED did not affect human chondrocytes viability under 10x concentration. Of interest, our data indicated that OleoGrape®SEED treatment significantly inhibited the IL-1ß-induced levels of the MMP-13, NO and PGE2 in isolated cultured chondrocytes (Fig. 1) as well as in the organ culture of human cartilage explants (Fig. 2). Moreover, sera collected from rabbits force-fed with extracts were found to exhibit an anti-IL-1ß effect in in vitro models of cultured chondrocytes (Fig. 3). X-ray of rabbits knees showed that OleoGrape®SEED decreased the ligament calcification, osteophytes formation, bone sclerosis and width of joint space narrowing associated
with a significant decrease in the radiographic score compared to ACLT vehicle group (Fig. 4).
To conclude, our data illustrated that OleoGrape®SEED counteracts the IL-1ß effects similarly in isolated chondrocytes and cartilage explants. Finally, HT and PCy were bioavailable and conserved their bioactivities in serum after oral consumption. We also provided in vivo evidence in post-traumatic OA model that OleoGrape®SEED exerts a modest but clearly noticeable effect on cartilage degradation. Taken together, these results suggest that OleoGrape®SEED may be of clinical significance for the OA treatment.
Figure 1: Effects of Oleogrape on IL-1ß induced MMP, NO and PGE2 production in human chondrocytes. Cells were pre-treated with 10x concentration of Oleogrape 24h prior to IL-1ß (1mg/mL) 24. Data were expressed as relative MMP-13, NO and PGE2 productions compared to control condition (1ng/mL). *p<0.05 compared with cells stimulated with IL-1ß.
Figure 2: Effects of Oleogrape on IL-1ß induced MMP, NO and PGE2 production in human cartilage explants.
Explants were pre-treated with 10x concentration of Oleogrape 24h prior to IL-1ß (1mg/mL) 24. Data were expressed as relative MMP-13, NO and PGE2 productions compared to control condition (1ng/mL). *p<0.05 compared with cells stimulated with IL-1ß.
Figure 3: Effects of sera from rabbits fed with Oleogrape on IL-1ß induced MMP, NO and PGE2 production in chondrocytes
Rabbits (n=3 per condition) were fed with Oleogrape (100mg/kg) or saline solution (NaCl) every two days during 6 days and serum was harvested 2h after the last gastric gavage. Cells were pre-treated with serum from rabbits at a final concentration of 2.5% (v/v) 24 h pripr to IL-1B (1ng/mL) 24h. Data were expressed as relative MMP-13, NO and PGE2 productions compared to control condition (1ng/mL). *p<0.05 compared with cells treated with serum from rabbits fed with NaCl and stimulated with IL-1ß.
Figure 4: Effects of Oleogrape on ACLT rabbits model. Rabbits (n=6 per group) underwent ACLT of the right knee. Vehicle (Veh) or Oleogrape was administered every two days for 13 weeks starting 3 weeks before surgery, X-ray and radiographic score of lateral control (Ct) or ACLT rabbits treated with vehicle or Oleogrape at 10 weeks following surgery. *p>0.05 compared to ACLT receiving vehicle group.
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