Demonstration of extracellular matrix protection factor-1’s disease-modifying properties as evidenced by radiographic analysis of the knee joint in a rat model of osteoarthritis
Location
Philadelphia, PA
Start Date
1-5-2024 1:00 PM
End Date
1-5-2024 4:00 PM
Description
Introduction: Osteoarthritis (OA) is a degenerative disease of the extracellular matrix(ECM). Extracellular Matrix Protection Factor (ECPF)-1 is a novel matrix metalloproteinase (MMP) inhibitor with possible application to the treatment of OA. ECPF-1 prevents the breakdown of the ECM by inhibiting MMP-1 and -13. Through the use of a rat model of OA, we investigated how ECPF-1 affects OA pathogenesis.
Objective: The primary objective was to determine the efficacy of ECPF-1 as a therapeutic that modulates the ECM degeneration associated with OA.
Methods: We utilized an infrapatellar injection of monoiodoacetate at week zero to model OA in rats. Animals were then treated with ECPF-1 (250nM - 5uM), saline injection (control) or BMP-7 (500ng) at days 7, 14, 21, and 28 and then sacrificed at day 28 for x-ray study. Within 30 days of the injection, OA morphology is apparent in this model. Hind limb (site of OA insult) distance stride during normal gait was used to measure mobility in the affected rats. Radiographic images of the rat hind limbs arranged at a 90-degree angle utilizing a measuring caliper were taken at 2x magnification using a kilovolt peak of 50kVp and milliamp seconds of 1 mAs. From the radiographs, measurements were obtained for M3 to visualize the shortest distance from the tibial cartilaginous surface to the femoral cartilaginous surface (joint space) and M4 for the distance from the outside of the femoral head to the posterior-most point of the calcaneus.
Results: Hind limb x-rays revealed a significantly increased distance between the tibial cartilaginous surface and the femoral cartilaginous surface (M3 = joint space) ( p<0.016 saline versus 250uM ECPF-1 one-way ANOVA with Brown-Forsythe test, and p<0.035 saline versus 250uM ECFP-1 Kriskal-Wallis post-test). Hind limb x-ray for the distance from the posterior edge of the femoral head to the posteriormost part of the calcaneus showed a statistically significant decrease in M4 length (p<0.035 saline versus 2.5uM ECPF-1 one-way ANOVA with Brown-Forsythe post-test and p<0.009 saline versus 2.5uM ECPF-1 Welch's T-test). The decrease in M4 length was supported by the decrease in average hind limb stride from 10 weeks to 12 weeks. The decrease in M4 length was also supported by showing a decrease in stride width from weeks 0 to 6.
Conclusion: Normalized gait and increased joint space after ECPF-1 treatment indicate the slowing of the progression of OA. Radiographic data supports the protection of cartilage when treated with ECPF-1, and this data is further supported by the normalization of gait in animals. This data is consistent with the decreased width of gait and improved mobility, supporting the idea that ECPF-1 protects cartilaginous breakdown in OA. Future studies will test the efficacy of ECPF-1 in target species such as canines or humans to validate this novel disease-modifying therapeutic for OA.
Embargo Period
5-23-2024
Demonstration of extracellular matrix protection factor-1’s disease-modifying properties as evidenced by radiographic analysis of the knee joint in a rat model of osteoarthritis
Philadelphia, PA
Introduction: Osteoarthritis (OA) is a degenerative disease of the extracellular matrix(ECM). Extracellular Matrix Protection Factor (ECPF)-1 is a novel matrix metalloproteinase (MMP) inhibitor with possible application to the treatment of OA. ECPF-1 prevents the breakdown of the ECM by inhibiting MMP-1 and -13. Through the use of a rat model of OA, we investigated how ECPF-1 affects OA pathogenesis.
Objective: The primary objective was to determine the efficacy of ECPF-1 as a therapeutic that modulates the ECM degeneration associated with OA.
Methods: We utilized an infrapatellar injection of monoiodoacetate at week zero to model OA in rats. Animals were then treated with ECPF-1 (250nM - 5uM), saline injection (control) or BMP-7 (500ng) at days 7, 14, 21, and 28 and then sacrificed at day 28 for x-ray study. Within 30 days of the injection, OA morphology is apparent in this model. Hind limb (site of OA insult) distance stride during normal gait was used to measure mobility in the affected rats. Radiographic images of the rat hind limbs arranged at a 90-degree angle utilizing a measuring caliper were taken at 2x magnification using a kilovolt peak of 50kVp and milliamp seconds of 1 mAs. From the radiographs, measurements were obtained for M3 to visualize the shortest distance from the tibial cartilaginous surface to the femoral cartilaginous surface (joint space) and M4 for the distance from the outside of the femoral head to the posterior-most point of the calcaneus.
Results: Hind limb x-rays revealed a significantly increased distance between the tibial cartilaginous surface and the femoral cartilaginous surface (M3 = joint space) ( p<0.016 saline versus 250uM ECPF-1 one-way ANOVA with Brown-Forsythe test, and p<0.035 saline versus 250uM ECFP-1 Kriskal-Wallis post-test). Hind limb x-ray for the distance from the posterior edge of the femoral head to the posteriormost part of the calcaneus showed a statistically significant decrease in M4 length (p<0.035 saline versus 2.5uM ECPF-1 one-way ANOVA with Brown-Forsythe post-test and p<0.009 saline versus 2.5uM ECPF-1 Welch's T-test). The decrease in M4 length was supported by the decrease in average hind limb stride from 10 weeks to 12 weeks. The decrease in M4 length was also supported by showing a decrease in stride width from weeks 0 to 6.
Conclusion: Normalized gait and increased joint space after ECPF-1 treatment indicate the slowing of the progression of OA. Radiographic data supports the protection of cartilage when treated with ECPF-1, and this data is further supported by the normalization of gait in animals. This data is consistent with the decreased width of gait and improved mobility, supporting the idea that ECPF-1 protects cartilaginous breakdown in OA. Future studies will test the efficacy of ECPF-1 in target species such as canines or humans to validate this novel disease-modifying therapeutic for OA.