ABSTRACT
In the context of modern drug discovery, there is an obvious advantage to designing phenotypic bioassays based on human disease-relevant cells that express disease-relevant markers. The specific aim of the study was to develop a convenient and reliable method for screening compounds with Tumor Necrosis Factor-alpha (TNF-α) inhibitory activity. This assay was developed using cryopreserved ready-to-use cartilage-derived cells isolated from juvenile donors diagnosed with polydactyly. It has been demonstrated that all donor (10 donors) cells were able to respond to TNF-α treatment by increased secretion of pro-inflammatory cytokine IL-6 into subcultural medium. Inhibition of TNF-α using commercially available TNF-α inhibitor etanercept resulted in a dose-dependent decrease in IL-6 production which was measured by Enzyme-Linked Immunosorbent Assay (ELISA). TNF-α dependent IL-6 production was detected in the cells after both their prolonged cultivation in vitro (≥20 passages) and cryopreservation. This phenotypic bioassay based on ready-to-use primary human cells was developed for detection of novel TNF-α inhibitory compounds and profiling of biosimilar drugs.
Subject(s)
Antirheumatic Agents/pharmacology , Arthritis, Rheumatoid/pathology , Cartilage/pathology , Etanercept/pharmacology , Tumor Necrosis Factor-alpha/antagonists & inhibitors , Antirheumatic Agents/therapeutic use , Arthritis, Rheumatoid/drug therapy , Arthritis, Rheumatoid/metabolism , Cartilage/drug effects , Cartilage/metabolism , Cells, Cultured , Child , Dose-Response Relationship, Drug , Drug Evaluation, Preclinical/methods , Etanercept/therapeutic use , Humans , Tumor Necrosis Factor-alpha/metabolismABSTRACT
The purpose of the present research was: 1. To investigate the opportunity of application of allogen osteoplastic materials such as Lioplast, received in Samara Tissue Bank of Samara State Medical University. 2. To work out a new technique of producing lyophilized allogen osteoplastic materials, such as Lioplast, with application of an ultrasonic method of clearing. 3. The development of various techniques of operative intervention on jaws for increase in volume of atrophied, as a result of secondary adentia, alveolar shoot. 4. The application of spongy allogen osteoplastic material as a skeleton for filling in surgical operations on jaws. 5. The demonstration of the Technique of a bone-periostic Lioplast 'sandwich' for formation of an alveolar shoot.
