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Summary Objective The goals of this study were (1) to quantify proteoglycan 4 ( PRG4 ) gene expression; (2) to assess lubricin immunostaining; and (3) to measure synovial fluid lubricin concentrations in clinical and experimental models of equine carpal osteoarthritis (OA). Design Lubricin synovial fluid concentrations and cartilage and synovial membrane PRG4 expression were analyzed in research horses undergoing experimental OA induction ( n = 8) and in equine clinical patients with carpal OA ( n = 58). Lubricin concentrations were measured using a custom sandwich enzyme-linked immunosorbent assay, and PRG4 expression was quantified using qRT-PCR. Lubricin immunostaining was assessed in synovial membrane and osteochondral sections in the experimental model. Results Lubricin concentrations increased in synovial fluid following induction of OA, peaking at 21 days post-operatively in OA joints vs sham-operated controls (331 ± 69 μg/mL vs 110 ± 19 μg/mL, P = 0.001). Lubricin concentrations also increased in horses with naturally occurring OA as compared to control joints (152 ± 32 μg/mL vs 68 ± 4 μg/mL, P = 0.003). Synovial membrane PRG4 expression increased nearly 2-fold in naturally occurring OA ( P = 0.003), whereas cartilage PRG4 expression decreased 2.5-fold ( P = 0.025). Lubricin immunostaining was more pronounced in synovial membrane from OA joints as compared to controls, with intense lubricin localization to sites of cartilage damage. Conclusions Although PRG4 gene expression decreases in OA cartilage, synovial membrane PRG4 expression, synovial fluid lubricin concentrations and lubricin immunostaining all increase in an equine OA model. Lubricin may be elevated to protect joints from post-traumatic OA.

Summary Autologous chondrocyte implantation (ACI) has improved outcome in long-term studies of joint repair in man. However, ACI requires sutured periosteal flaps to secure the cells, which precludes minimally-invasive implantation, and introduces complications with arthrofibrosis and graft hypertrophy. This study evaluated ACI on a collagen type I/III scaffold (matrix-induced autologous chondrocyte implantation; MACI ® ) in critical sized defects in the equine model. Methods Chondrocytes were isolated from horses, expanded and seeded onto a collagen I/III membrane (ACI-Maix™) and implanted into one of two 15-mm defects in the femoral trochlear ridge of six horses. Control defects remained empty as ungrafted debrided defects. The animals were examined daily, scored by second look arthroscopy at 12 weeks, and necropsy examination 6 months after implantation. Reaction to the implant was determined by lameness, and synovial fluid constituents and synovial membrane histology. Cartilage healing was assessed by arthroscopic scores, gross assessment, repair tissue histology and immunohistochemistry, cartilage glycosaminoglycan (GAG) and DNA assay, and mechanical testing. Results MACI ® implanted defects had improved arthroscopic second-look, gross healing, and composite histologic scores, compared to spontaneously healing empty defects. Cartilage GAG and DNA content in the defects repaired by MACI implant were significantly improved compared to controls. Mechanical properties were improved but remained inferior to normal cartilage. There was minimal evidence of reaction to the implant in the synovial fluid, synovial membrane, subchondral bone, or cartilage. Conclusions The MACI ® implant appeared to improve cartilage healing in a critical sized defect in the equine model evaluated over 6 months.

SummaryChondrocyte-collagen composites were evaluated for resurfacing of large articular defects. Isolated chondrocytes were cultured in expanded collagen scaffolds for 7–10 days to provide a composite containing 3.6 × 104 cells/mm3. The graft was transplanted into 15 mm full thickness articular defects in the femoropatellar joint of 12 horses using arthroscopic techniques. Ungrafted defects in the opposite femoropatellar joint served as controls. Synovial fluid, clinical progress and pain responses were evaluated in groups of 6 horses over 4-month and 8-month periods. Following termination, gross, histochemical and histologic evaluations of the repair tissues and synovial membrane were performed. Arthroscopic defect debridement and chondrocyte implantation resulted in minimal post-operative effusion or pain, and synovial fluid constituents were not significantly different in grafted and ungrafted joints. Gross differences in grafted defects were not apparent. Increased chondrocyte numbers and cartilage histochemical staining were evident in the deeper layers of grafted defects, whereas ungrafted defects were almost entirely fibrous tissue. The surface layers of grafted defects were fibrous tissue. There were no synovial fluid cellular responses, synovial membrane histiocytic reaction or subchondral bone infiltrates to suggest immune-related reaction to the allograft cells. Chondrocyte-collagen grafts were arthroscopically implanted and resulted in improved cartilage healing in extensive defects. However, the structural organization of the surface layers was inadequate and suggested poor long-term durability.

Chondrocyte-laden collagen scaffolds were evaluated in extensive cartilage defects in an equine model. Arthroscopic techniques were used to implant a chondrocyte-collagen culture product in 15-mm defects in the lateral trochlear ridge of the femoropatellar joint of 12 horses. Ungrafted control defects were formed in the opposite joint. Groups of six horses were terminated at 4 and 8 months after implantation and the repair sites, adjacent cartilage, and remote cartilage within each femoropatellar joint examined biochemically. Eight months following surgery the relative proportions of type II collagen in grafted and ungrafted defects, determined using the ratio of cyanogen bromide cleavage products alpha 1(II)CB10/alpha 2(I)CB3,5, were not significantly different (31.57 +/- 2.76% and 26.88 +/- 2.76%, respectively). Aggrecan content was significantly improved in grafted defects (85.61 +/- 6.51 and 74.91 +/- 10.31 micrograms/mg dry weight). Cartilage surrounding grafted defects also showed improved maintenance of cartilage glycosaminoglycan content. Thus, chondrocyte grafting in collagen scaffold vehicles improved the aggrecan content in extensive cartilage defects and surrounding normal cartilage. However, given the continued disparity between repair tissue and normal cartilage aggrecan content, and the low proportion of type II collagen in grafted defects, the utility of collagen scaffolds for chondrocyte grafting of large cartilage defects seems limited.