In human knee articular chondrocytic-spheroids, Orthogrit modulates TNF-α and IL-1β induced inflammation, oxidative stress, ECM catabolism, and improves locomotory behaviour in Caenorhabditis elegans

Abstract

Osteoarthritis is a progressive chronic degenerative joint disease characterized by inflammation, pain, and articular cartilage deterioration. The current pharmacotherapies for osteoarthritis do not address the progression of osteoarthritis and are also associated with several adverse effects. Herbal medicines are emerging as potential source for the safe and effective treatment of osteoarthritis. Orthogrit, an herbo-mineral prescription medicine, has anti-oxidant, anti-inflammatory, and cartilage promoting properties that can potentially halt progression of osteoarthritis by its multifaceted bioactivities. The present study aimed to characterize the pharmacological effects of Orthogrit using 3D culture of human knee articular chondrocytes (NHAC-kn) and Caenorhabditis elegans. The chemical characterization of Orthogrit was performed by UHPLC analysis. The in vitro evaluation of Orthogrit was performed on TNF-α and IL-1β co-induced NHAC-kn spheroids; and in vivo analysis on LPS- exposed N2 (wild-type) strain of C. elegans. In chondrocytic spheroids, Orthogrit treatment reduced chondrotoxicity, ROS levels, and release of IL-6, PGE2, and CTXII, a marker of cartilage degradation. Treatment with Orthogrit normalized mitochondrial membrane potential; levels of aggrecan, and sulphated glycosaminoglycans; and reporter activity of IL-1β and NF-κB. In LPS-exposed C. elegans, Orthogrit treatment decreased nematode mortality, ROS levels and normalized locomotory behaviour (reversal and omega turns), SOD, Catalase and GSH levels. The mRNA expression analysis revealed that Orthogrit acts against progression of osteoarthritis by regulating inflammation mediators (JAK2, COX2), catabolic ECM enzymes (MMP1, MMP3, ADAMTS-4), redox homeostasis (Nrf2), p38 MAPK signalling (PMK-1, SEK-1) and apoptosis (CED-3). Taken together, Orthogrit is a potential therapeutic agent for management of osteoarthritis.