Accepted for/Published in: JMIR Research Protocols
Date Submitted: Sep 26, 2022
Date Accepted: May 5, 2023
Warning: This is an author submission that is not peer-reviewed or edited. Preprints - unless they show as "accepted" - should not be relied on to guide clinical practice or health-related behavior and should not be reported in news media as established information.
Creating a Proinflammatory Chondrocyte Cell Model to Investigate Anti-inflammatory Leads: A Proof-of-Concept Investigation
ABSTRACT
Background:
In vitro cell models are pivotal in medicine and biology because they provide insight into cells' biochemical pathways, simulating bioactivities, mechanics and physiological behavior of organs or organ systems. In this study, we have optimized a protocol for a proinflammatory chondrocyte model.
Objective:
In this study, a process was delineated for the easy regeneration of chondrocytes to develop an inflammatory model. Since, inflammation in chondrocytes is a key phenomenon involved in the development and progression of OA, an in vitro model was established in this study to investigate the anti-inflammatory properties of vitamin D.
Methods:
Chondrocytes were grown from bone marrow mesenchymal stem cells (BMSC). Inflammation was induced in these chondrocytes using lipopolysaccharides (LPS) (large molecules consisting of a lipid and a polysaccharide). To assess inflammation, we measured the levels of proinflammatory marker - tumor necrosis factor-α (TNFα), in the LPS treated chondrocytes using ELISA. Additionally, we also appraised the effect of vitamin – D on the proinflammatory chondrocyte model to assess if this model can be employed for the investigation of therapeutic leads.
Results:
Chondrocytes successfully differentiated from BMSCs. Expression of TNFα was elevated in the LPS treated chondrocytes in comparison to the control (chondrocytes treated with buffer). Vitamin D attenuated the expression of TNF α in the LPS treated chondrocytes indicating that the in vitro proinflammatory cell model can be used for the investigation of therapeutic leads.
Conclusions:
Chondrocytes are generally isolated for different physiochemical investigations from cartilage tissue. Because of limited availability of cartilage and low yield of viable chondrocytes from cartilage, such physiochemical investigations often prove difficult and expensive. The protocol depicted in this study can not only be employed to generate large number of viable chondrocytes but also can be used for evaluating the effect of therapeutic leads towards chondrocytes’ inflammatory response, and therefore may prove valuable for investigating, managing, and treating diseases such as rheumatoid arthritis and osteoarthritis. Clinical Trial: This study does not require trial registration.
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