IntroductionCartilage has a limited capacity for self-repair due in part to its avascular nature, making proper mechanical loading and nutritional support essential for maintaining joint health. Nutraceuticals derived from agricultural by-products present an opportunity to enhance animal health while retaining product value within the production system. Hemp polyphenol extract, a product derived from a by-product of hemp plant processing, contains bioactive compounds such as cannabidiol (CBD) and flavonoids that have been associated with anti-inflammatory and chondroprotective effects. The objective of this study was to investigate the potential anti-inflammatory and/or chondroprotective properties of a simulated digest of hemp polyphenol extract (HPE) on porcine cartilage explants challenged with lipopolysaccharide (LPS). MethodsTwo experiments were conducted to determine non-cytotoxic, effective concentrations of HPE. Cartilage explants were aseptically collected from the intercarpal joints of market-weight pigs (Experiment 1: n = 9; Experiment 2: n = 12) and cultured at 37 °C and 7% CO2 for 120 h. During the final 96 h, explants were conditioned with varying concentrations of HPE (1.22, 2.44, 7.32, 12.2, or 24.4 mg/mL) and stimulated with LPS (0 or 10 μg/mL) during the final 48 h. Samples of culture media were collected prior to LPS stimulation and at 24 and 48 h post-stimulation for analysis of inflammatory biomarkers [prostaglandin E2 (PGE2), nitric oxide (NO), and resolvin D1 (RvD1)]. Cartilage structural integrity was assessed by quantifying glycosaminoglycan release into media (mGAG) and retention within explants (tGAG), and explants were stained to evaluate cell viability.ResultsIn Trial 1, HPEtreatments (1.22, 2.44, and 24.4 mg/mL) did not markedly alter inflammatory biomarkers but reduced cell viability at the highest dose (24.4 mg/mL). Based on these results, Trial 2 was conducted using intermediate concentrations (2.44, 7.32, and 12.2 mg/mL). Conditioning of explants with 7.32 and 12.2 mg/mL significantly reduced PGE2 and NO concentrations compared to controls, while RvD1 was elevated across all treatments, with the highest concentrations observed at 12.2 mg/mL across all time points. At 12.2 mg/mL mGAG was decreased while tGAG was increased. Cell viability remained above 99% in all treatment groups. DiscussionThese findings indicate that intermediate doses of HPE may provide anti-inflammatory and chondroprotective benefits, supporting the potential use of hemp by-products as functional nutraceutical ingredients for joint health.
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