Toward Development of a Diabetic Synovium Culture Model

Neeraj Sakhrani; Andy J Lee; Lance A Murphy; Hagar M Kenawy; Christopher J Visco; Gerard A Ateshian; Roshan P Shah; Clark T Hung

doi:10.3389/fbioe.2022.825046

Toward Development of a Diabetic Synovium Culture Model

Front Bioeng Biotechnol. 2022 Feb 21:10:825046. doi: 10.3389/fbioe.2022.825046. eCollection 2022.

Authors

Neeraj Sakhrani¹, Andy J Lee¹, Lance A Murphy¹, Hagar M Kenawy¹, Christopher J Visco², Gerard A Ateshian^{1

3}, Roshan P Shah⁴, Clark T Hung^{1

4}

Affiliations

¹ Department of Biomedical Engineering, Columbia University, New York, NY, United States.
² Department of Rehabilitation and Regenerative Medicine, Columbia University, New York, NY, United States.
³ Department of Mechanical Engineering, Columbia University, New York, NY, United States.
⁴ Department of Orthopedic Surgery, Columbia University, New York, NY, United States.

Abstract

Osteoarthritis (OA) is a degenerative joint disease characterized by articular cartilage degradation and inflammation of synovium, the specialized connective tissue that envelops the diarthrodial joint. Type 2 diabetes mellitus (DM) is often found in OA patients, with nearly double the incidence of arthritis reported in patients with diabetes (52%) than those without it (27%). The correlation between OA and DM has been attributed to similar risk factors, namely increasing age and joint loading due to obesity. However, a potential causative link is not well understood due to comorbidities involved with treating diabetic patients, such as high infection rates and poor healing response caused by hyperglycemia and insulin resistance. The purpose of this study was to investigate the effect of hyperglycemic and insulin culture conditions on synovium properties. It was hypothesized that modeling hyperglycemia-induced insulin resistance in synovium would provide novel insights of OA pathogenesis in DM patients. To simulate DM in the synovial joint, healthy synovium was preconditioned in either euglycemic (EG) or hyperglycemic (HG) glucose concentrations with insulin in order to induce the biological response of the diseased phenotype. Synovium biochemical composition was evaluated to determine ECM remodeling under hyperglycemic culture conditions. Concurrent changes in AKT phosphorylation, a signaling pathway implicated in insulin resistance, were measured along with gene expression data for insulin receptors, glucose transporters, and specific glycolysis markers involved in glucose regulation. Since fluid shear stress arising during joint articulation is a relevant upstream stimulus for fibroblast-like synoviocytes (FLS), the predominant cell type in synovium, FLS mechanotransduction was evaluated via intracellular calcium ([Ca²⁺]_i). Incidence and length of primary cilia, a critical effector of cell mechanosensing, were measured as potential mechanisms to support differences in [Ca²⁺]_i responses. Hyperglycemic culture conditions decreased collagen and GAG content compared to EG groups, while insulin recovered ECM constituents. FLS mechanosensitivity was significantly greater in EG and insulin conditions compared to HG and non-insulin treated groups. Hyperglycemic treatment led to decreased incidence and length of primary cilia and decreased AKT phosphorylation, providing possible links to the mechanosensing response and suggesting a potential correlation between glycemic culture conditions, diabetic insulin resistance, and OA development.

Keywords: calcium signaling; hyperglycemia; insulin resistance; osteoarthritis; primary cilia; shear stress; synovium; type 2 diabetes.