Monosodium iodoacetate

Rats were randomised and divided into two groups based on the OA model (subacute/chronic arthritis model, n = 50 rats per group) and further divided into four treatment groups (n = 6 rats per group). Monosodium iodoacetate (Sigma) was dissolved in 0.9% normal saline. With rats under anaesthesia by isoflurane inhalation, their left knee joints received a single intra-articular injection of 3 mg MIA in a volume of 50 μL using a 30-G needle. Control rats were injected with an equivalent volume of normal saline. hASC-EVs (1 × 10⁸ particles) were given in a 30 μL volume per joint. PBS and hyaluronic acid (HA, 2.67 × 10⁶ Da, Lifecore biomedical, MN, SUA, 0.3 mg) were injected under the same condition for each group. Rats in the subacute arthritis group were treated with EVs and HA once a week for 21 days before significant OA progression (1 week after OA induction). In the chronic arthritis group, treatments were given twice a week for 40 days after significant OA progression (2 weeks after OA induction). After 28 days (subacute arthritis) and 56 days (chronic arthritis) of OA induction, rats were sacrificed by CO₂ inhalation, and the knee joints of each group were dissected and the surrounding muscle was trimmed for further analyses.

Commercial PX injection (Rheoma^® injection, Samsung Pharmaceuticals, 20 mg/ml as PX) for IA and IM use was obtained from the local hospital pharmacy. Sodium hyaluronate of microbial origin (molecular weight: 1500–2500 kDa) was purchased from Humedix Co., Ltd (Sungnam, Korea). Mo nosodium iodoacetate and isoxicam were purchased from Sigma-Aldrich (St Louis, MO, USA). Acetonitrile, methanol, formic acid, and deionized water were obtained from J. T. Baker (Phillipsburg, NJ, USA). All other chemicals used were of the highest commercial grade available.

The animals were randomized and assigned to three groups (n=10/per group) as follows: Sham group, OA model group and resveratrol treatment group. Following anesthetization with isoflurane (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany), rats in the OA model group and the resveratrol treatment group were injected with intra-articular injection of monosodium iodoacetate (Sigma-Aldrich; Merck KGaA) through the patellar ligament into the intra-articular space of the right knee. Rats in the resveratrol treatment group were administered with 50 mg/kg/3 days resveratrol, for 8 weeks. Rats were sacrificed using decollation under 35 mg/kg pentobarbital sodium (Sigma-Aldrich; Merck KGaA), after treatment with resveratrol.

Leaves of W. fruticosa were collected in the month of October from ‘Gharsi’ village hills, Solan Himachal Pradesh, India. Plant was authenticated at the department of forestry, Dr. Y.S. Parmar University Solan, India and linked to UHF-Herbarium with field book number 12545. Carrageenan, Freund’s adjuvant and monosodium iodo acetate were procured from Sigma Aldrich, Bangalore Genei. TNF-α was estimated using commercial ELISA kit procured from Genxbio, Delhi. All other chemicals used in present investigation were of analytical grade.