Abstract
Background: Losac is a hemolin from Lonomia obliqua caterpillar that promotes blood coagulation through proteolytic activation of factor X. In HUVECs, Losac increased cell proliferation and inhibited the apoptosis induced by starvation. Additionally, we have observed the cytoprotective effect of rLosac in other cell types, such as neurons and fibroblasts. Recombinant Losac (rLosac) was obtained from bacteria system. This report aims to investigate the potential of rLosac as a chondroprotective agent in human articular chondrocytes and to analyze its capacity to stimulate the production of extracellular matrix molecules and to reduce senescence. Methods: Primary cultures of human chondrocytes were prepared from cartilage of osteoarthritic knee joints obtained from 4 donors (age range: 68‐83) undergoing total knee replacement surgery. The study was carried out in full accordance with local ethics guidelines, and cartilage samples were collected after obtaining written informed consent of the donors. Chondrocytes were treated with rLosac and several parameters were measured: specific senescence‐associated beta‐galactosidase activity, cell viability by the MTT assay, production of extracellular matrix molecules by western blot and immunofluorescence microscopy, and cell cycle by flow cytometry. Results: rLosac was found to be an effective anti‐apoptotic agent, as evidenced by the strong inhibition of cellular death. The cytoprotective effect was observed specially starting from 48h and 72h of treatment, even in the presence of IL‐1β (50 ng/ml) to induce an inflammatory environment. Moreover, rLosac was able to reduce the expression of β‐galactosidade after 6 days of treatment. In addition, rLosac stimulated the production of collagen type I, laminin and fibronectin, and to increase the expression of the receptor of collagen type I, and to decrease the expression of IL‐1β. Conclusion: rLosac has the potential to become a therapeutic agent for protection of articular cartilage against the progression of osteoarthritic diseases.Grant Funding Source: CETICs/FAPESP (Processo No. 2013/07467‐1), CAPES, INCTTox‐CNPq
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