Purpose: Osteoarthritis (OA) is one of the most common and debilitating joint diseases. This age-related disease has currently no effective treatment, and the diagnosis in human is based on pain and functional symptoms, and can be confirmed by radiography. Most current preclinical studies use histological analyses to evaluate new therapies, which remains a time-consuming and laborious technique and cannot be used in the human clinical context. Interestingly, non-invasive techniques allowing a longitudinal analysis of osteoarthritis in animal models have also been set up, such as imaging techniques (radiography, MRI, micro-CT) or gait analysis. In this context, the aim of the present work was to assess the anti-osteoarthritic effect of a combination of natural molecules (ie green tea extract, collagen hydrolysates and a mix of chondroitin/glucosamine) in vivo by concomitantly evaluating OA progression in a spontaneous murine model by two non-invasive methods (X-ray and gait analysis). Methods: The in vitro effect of the cocktail, which consisted of 250 μg/ml of green tea extract, 0,5 mg/ml of collagen hydrolysates and 200 μg/ml of chondroitin sulfate mixed with 5% glucosamine sulfate, on the inflammation-associated OA events was first evaluated on IL1-β-treated rabbit articular chondrocytes. The expression levels of aggrecan, type II collagen and Sox9 transcripts, which are typical chondrogenic markers, have been evaluated after 3 days of culture by RT-qPCR. The prostaglandin-E2 (PGE2) production, as well as iNOS, COX2 and MMP13 transcript levels have also been determined after 24h of treatment with IL1-β (10 ng/ml) in the presence of the cocktail. Extracellular matrix production was also estimated using a glycosaminoglycan (GAG) staining with Alcian Blue after 14 days of treatment. In vivo, OA progression was evaluated in 18-month-old C57BL/6 mice used as a model of spontaneous osteoarthritis. To monitor the progression of the disease and appreciate the effects of a 4 month treatment with the cocktail given orally as an enriched diet containing green tea extract (300 mg/kg), collagen hydrolysates (100 mg/kg) and chondroitin sulfate mixed with 5% glucosamine sulfate (200 mg/kg), two non-invasive methods were used: a radiological OA scoring and a functional gait analysis using the CatWalk® system. Results: Our results show a marked reduction of the IL1-β-mediated production of PGE2 associated with an inhibition of the expression levels of COX2 and iNOS transcripts in cocktail-treated versus untreated cells. The production of the catabolic enzymes MMP13 was also found to be drastically inhibited. Our data also indicate that our cocktail induced a significant increase in the levels of chondrogenic markers (aggrecan, type II collagen and Sox9) and GAG production. In vivo, our results show a global increase in the radiological OA score during the progression of the disease, as well as anomalies in mice gait notably evidenced by an increase in the Max Contact parameter evaluated with the CatWalk® system. After 4 months of treatment with the cocktail-enriched diet, we found a significant decrease in the radiological OA score, and a significant decrease in the Max Contact parameter between treated and untreated mice. Conclusions: Our data demonstrate that a combination of natural molecules exerts anti-IL-1-β effects on cultured chondrocytes. This work also demonstrates that a longitudinal analysis of OA progression in mice is feasible by concomitantly using radiology and gait analysis, thereby mimicking the human clinical practice. Interestingly, we also demonstrated the in vivo anti-osteoarthritic effects of a natural molecules-containing cocktail by measuring clinically relevant parameters with radiology and gait analysis. Taken together, our data may help pave the way of promising therapeutic approaches based on enriched diets with natural molecules.