POS0404 VOLUNTARY WHEEL RUNNING MODEL IN MICE TO MECHANICALLY STIMULATE THE ENTHESIS OF THE ACHILLES TENDON
Authors: A. Briolay, S. Delplace, F. Duboeuf, O. Peyruchaud, D. Magne, L. Brizuela, C. Bougault
Journal: Annals of the Rheumatic Diseases (2021)

Background: Excessive bone formation in the entheses is one of the features of peripheral spondyloarthritis. Biomechanical stress is proposed to occupy a central place in spondyloarthritis pathophysiology, but the precise molecular and cellular mechanisms underlying the pathological response of the enthesis are still largely unknown [1]. Besides, physical therapy and exercise are recommended as non-pharmacologic therapies for patients. We focused on the effect of exercising on enthesis ossification. Objectives: We aimed to develop and characterize an in vivo model in mice to study the impact of mechanical stimulation on the enthesis of the Achilles tendon. Methods: DBA/1 mice were subjected to voluntary running exercise by the use of activity wheels for two weeks, and compared to mice housed in standard conditions (n=17 per group). The running performances were recorded. mRNAs were extracted from the long bones (flushed tibia and femur) and the ankles’ entheses for real-time PCR analysis. µCT was performed on the femurs. Alkaline phosphatase activity was detected by histology on the anchorage of the Achilles tendon to the calcaneum, and by enzymatic assay in serum samples. Luminex analysis was also conducted on serum samples for Il-6 and Il-8/Kc detection. Results: Free access to the activity wheel resulted in a running exercise of 5.5±0.8 km/day (approximately 80 km in total) at 14.5±0.5 m/min. No effect was detected on the femur architecture by µCT. Sclerostin ( Sost ) gene expression was monitored as a mechanosensitive marker. Its expression was expectedly reduced by half in entheseal tissues, but no modulation was observed in long bones (Figure 1). Similarly, exercise-induced regulation of Osterix and Runx2 expressions was observed only in enthesis samples. This tissue-specific pattern was also verified for key genes of the sphingosine-1 phosphate metabolic pathway, which we recently implicated in spondyloarthritis pathophysiology [2]. The in situ staining of alkaline phosphatase activity suggested the presence of more positive cells in the anchorage of Achilles tendon of running mice, compared to control ones. However, alkaline phosphatase activity in serum samples and its gene expression in rough tissue extracts were unchanged. No inflammatory response was detected as Il-8/Kc serum levels were similar in the control and the exercising group (59±14 vs 57±14 pg/mL). In addition, Il-6 was not detected in the serum and its expression was very faint and constant in the tissue extracts. Conclusion: This work is still in progress for a more complete characterization of the model. We believe that this experimental design will be useful to study the role of mechanical stimulation specifically in the enthesis and that it can help to better understand the spondyloarthritis pathophysiology. References: [1]Cambré, et al. Nat Commun , 2018; [2] El Jamal, et al. J Bone Miner Res , 2019 Figure 1. Expression level of the mechanosensitive gene Sclerostin (Sost). It dropped in response to exercise in entheseal tissues, but not in long bones, revealing a tissue-specific response to mechanical stimulation. Acknowledgements: Société Arthritis R&amp;D (2020) Disclosure of Interests: None declared