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Articular cartilage defects are a common source of joint pain and dysfunction. We hypothesized that sustained low-dose dexamethasone (DEX) delivery via an acellular osteochondral implant would have a dual pro-anabolic and anti-catabolic effect, both supporting the functional integrity of adjacent graft and host tissue while also attenuating inflammation caused by iatrogenic injury. An acellular agarose hydrogel carrier with embedded DEX-loaded poly(lactic-co-glycolic) acid (PLGA) microspheres (DLMS) was developed to provide sustained release for at least 99 days. The DLMS implant was first evaluated in an in vitro pro-inflammatory model of cartilage degradation. The implant was chondroprotective, as indicated by maintenance of Young's modulus (EY) (p = 0.92) and GAG content (p = 1.0) in the presence of interleukin-1β insult. In a subsequent preliminary in vivo experiment, an osteochondral autograft transfer was performed using a pre-clinical canine model. DLMS implants were press-fit into the autograft donor site and compared to intra-articular DEX injection (INJ) or no DEX (CTL). Functional scores for DLMS animals returned to baseline (p = 0.39), whereas CTL and INJ remained significantly worse at 6 months (p Statement of significance Articular cartilage defects are a common source of joint pain and dysfunction. Effective treatment of these injuries may prevent the progression of osteoarthritis and reduce the need for total joint replacement. Dexamethasone, a potent glucocorticoid with concomitant anti-catabolic and pro-anabolic effects on cartilage, may serve as an adjuvant for a variety of repair strategies. Utilizing a dexamethasone-loaded osteochondral implant with controlled release characteristics, we demonstrated in vitro chondroprotection in the presence of IL-1-induced degradation and improved in vivo functional outcomes following osteochondral repair. These improved outcomes were correlated with superior histological cartilage scores and minimal-to-no comorbidity, which is a risk with high dose dexamethasone injections. Using this model of cartilage restoration, we have for the first time shown the application of targeted, low-dose dexamethasone for improved healing in a preclinical model of focal defect repair.

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.

Little is known about the critical role of the synovium in joint homeostasis and osteoarthritis (OA). We describe a novel in vitro tissue-engineered (TE) model to investigate the structure–function of synovium through quantitative solute transport measures. This TE synovium model was developed using healthy bovine, CD14(−) fibroblast-like synoviocytes (FLS, or Type B synoviocytes) encapsulated in a Matrigel scaffold. The bovine system is well established in musculoskeletal research, allowing comparisons to be made to previous work. Sheet-like TE constructs were precultured to attain native protein composition and polarized structure, as determined by immunohistochemistry and confocal microscopy, and subsequently exposed to interleukin-1α (IL) or dexamethasone (DEX). The biological responses, including nitric oxide and hyaluronic acid (HA) secretion, of engineered synovium paralleled that of native synovium. While HA media content increased in response to both IL and DEX, a higher proportion of HA was low molecular weight (

Tissue engineering approaches to treatment of intervertebral disc degeneration (IDD) represent a novel avenue of addressing the biologic basis of this disease. However, such approaches remain limited by their invasive nature and disruption to the annular fibrosis (AF). This study sought to explore a new minimally-invasive tissue-engineering approach utilizing an injection of a photopolymerizable biogel scaffold seeded with mesenchymal stem cells (MSCs) directly into the nucleus pulposus (NP). This study was performed using rabbit specimens for both in vivo and in vitro outcome measures. The treatment in this study was performed by injecting 25 μl of 10% (w/v) methacrylated gelatin biogel with 0.15% (w/v) lithium phenyl 2,4,6-trimethylbenzoylphosphinate (LAP) and rabbit MSCs (1 × 106 ) cells/ml into the NP. Samples were then photopolymerized in situ using non-ultraviolet light irradiation via a fiberoptic wire. For the in vitro arm of this study, gene expression analysis demonstrated increased anabolic activity in irradiated MSCs with and without biogel scaffolds. For the in vivo arm of this study, while GAG analysis did not demonstrate significant differences between groups, MRI analysis exhibited a trend toward improved NP matrix. Histological analysis was consistent with increased cellularity and less severe disc degeneration in the MSC + Gel group. However, osteophyte formation was noted in both Stab and MSC + Gel groups after the study period. Increased matrix gene expression of irradiated groups within in vitro studies indicates a photobiologic effect of 405 nm light. Despite promising anabolic actions, osteophyte formation and AF defects could not be avoided with implementation of this minimally-invasive tissue-engineering approach. © 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:1451-1459, 2019.

The synovium plays a key role in the development of osteoarthritis, as evidenced by pathological changes to the tissue observed in both early and late stages of the disease. One such change is the attachment of cartilage wear particles to the synovial intima. While this phenomenon has been well observed clinically, little is known of the biological effects that such particles have on resident cells in the synovium. The present work investigates the hypothesis that cartilage wear particles elicit a pro-inflammatory response in diseased and healthy human fibroblast-like synoviocytes, like that induced by key cytokines in osteoarthritis. Fibroblast-like synoviocytes from 15 osteoarthritic human donors and a subset of three non-osteoarthritic donors were exposed to cartilage wear particles, interleukin-1α or tumor necrosis factor-α for 6 days and analyzed for proliferation, matrix production, and release of pro-inflammatory mediators and degradative enzymes. Wear particles significantly increased proliferation and release of nitric oxide, interleukin-6 and -8, and matrix metalloproteinase-9, -10, and -13 in osteoarthritic synoviocytes, mirroring the effects of both cytokines, with similar trends in non-osteoarthritic cells. These results suggest that cartilage wear particles are a relevant physical factor in the osteoarthritic environment, perpetuating the pro-inflammatory and pro-degradative cascade by modulating synoviocyte behavior at early and late stages of the disease. Future work points to therapeutic strategies for slowing disease progression that target cell-particle interactions. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:1979-1987, 2019.