Granzyme-A deficiency attenuates experimental osteoarthritis in mice, but perforin deficiency does not | Online First

Jorge Calvo^1#, Llipsy Santiago^2,#, Maykel Arias², Julián Pardo^2,3,4, Jorge Albareda^1,2,5, Luis Martínez-Lostao^2,3,6*, Felícito García-Alvarez^1,2,5*

¹Department of Orthopaedic Surgery and Traumatology, Hospital Clínico “Lozano Blesa”, Zaragoza, Spain
²Instituto de Investigación Sanitaria Aragón (IIS Aragón), Zaragoza, Spain
³Department of Microbiology, Pediatrics, Radiology and Public Health, University of Zaragoza, Zaragoza, Spain
⁴CIBER de Enfermedades Infecciosas (CIBERINFEC), Madrid, Spain
⁵Department of Surgery, University of Zaragoza, Zaragoza, Spain
⁶Department of Immunology, Hospital Clínico “Lozano Blesa”, Zaragoza, Spain

Keywords: Granzyme, osteoarthritis, perforin.

Abstract

Objectives: This study aims to assess the development of osteoarthritis (OA) in granzyme A- (gzmA) and B- (gzmB) and perforin- (perf) knockout mice.

Materials and methods: A total of 75 male and female C57BL/6 (eight to nine-week-old) mice were allocated to: gzmA-deficient (gzmA-/-) (11 females, 8 males), gzmB-deficient (gzmB-/-) (9 females, 8 males), perf-deficient (perf-/-) (10 females, 9 males), and control group (10 females, 10 males). Osteoarthritis was induced in the right knee by instability of the meniscus medial ligament. Sham surgery was practiced in the left knee. Knee samples obtained eight weeks after surgery were stained (Safranin-O) and blindly scored in lateral and medial femur and tibia using the Osteoarthritis Research Society International scale (OARSI) (from Grade 0, cartilage intact to 6, deformation), (five stages from 0, no OA to 4, >50% surface involvement); OARSI score (grade x stage); and a semi-quantitative scale from Grade 0 (normal) to 6 (cartilage erosion >80%).

Results: Significantly higher values in all scales in the right knees compared to the left knees in male and female mice were observed (p<0.05). Males of all strains showed in the right knee higher values than females on all scales. Deficiency of perforin did not modify OA severity in any sex. The gzmA-/- females presented less degenerative changes than the other groups.

Conclusion: Our study results show that sex plays an important role in the development of experimental OA in mice. Deficiency of gzmA can protect from the development of OA in female mice.

# These authors share first authorship

* These authors share corresponding and senior authorship.

Citation: Calvo J, Santiago L, Arias M, Pardo J, Albareda J, Martínez-Lostao L, García-Alvarez F. Granzyme-A deficiency attenuates experimental osteoarthritis in mice, but perforin deficiency does not. Jt Dis Relat Surg 2023;34(2):i-viii.