Granzyme-A deficiency attenuates experimental osteoarthritis in mice, but perforin deficiency does not
Jorge Calvo1#
, Llipsy Santiago2,#, Maykel Arias2, Julián Pardo2,3,4, Jorge Albareda1,2,5, Luis Martínez-Lostao2,3,6*, Felícito García-Alvarez1,2,5*
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):271-278.
All experimental procedures were approved by the Animal Care and Research Committee of the University of Zaragoza (Spain) (PI27/17) comply with the ARRIVE guidelines and were carried out in accordance with the U.K. Animals (Scientific Procedures) Act, 1986 and associated guidelines and EU Directive 2010/63/EU for animal experiments.
Conceptualization, methodology, software, data curation, formal analysis, writing-original draft preparation, writing-reviewing and editing: J.C.; Visualization, methodology, data curation, formal analysis, investigation, writing-reviewing and editing: L.S.; Visualization, data curation, investigation, writingreviewing and editing: M.A.; Conceptualization, resources, supervision, formal analysis, writing-reviewing and editing: J.P.; Visualization, formal analysis, writing-reviewing and editing: J.A.; Conceptualization, supervision, formal analysis, writing-original draft preparation, writing-reviewing and editing: L.M.L.; Conceptualization, methodology, investigation, supervision, formal analysis, writing-original draft preparation, writing-reviewing and editing: F.G.A.
The authors have received economic help from Fondo Europeo de Desarrollo Regional, Gobierno de Aragón, Group B29_17R, Fundación Santander-Universidad de Zaragoza (Programa COVID-19), Agencia Estatal de Investigación 564 (PID2020-113963RBI00), Fundación Inocente, ASPANOA and Carrera de la Mujer de Monzón. LS is supported by a PhD fellowship (FPI) and by a postdoctoral fellowship 'Juan de la Cierva- Formación from the Ministry of Science, Innovation and Universities MA is supported by a postdoctoral fellowship 'Juan de la Cierva- incorporacion' from the Ministry of Science, Innovation and Universities. JP is supported by the ARAID Foundation.
The authors received no financial support for the research and/or authorship of this article.
The authors would like to thank the Servicios Científico Técnicos (IACS) and Servicio de Apoyo a la Investigación (Universidad de Zaragoza). Work in the JP laboratory is funded by the FEDER (Fondo Europeo de Desarrollo Regional, Gobierno de Aragón, Group B29_2R), grant PID2020-113963RBI00 by MICIN/AEI and CIBER -Consorcio Centro de Investigación Biomédica en Red- (CIBERINFEC, CB21/13/00087), Instituto de Salud Carlos III. LS is supported by a PhD fellowship (FPI) and by a postdoctoral fellowship 'Juan de la Cierva- Formación from the Ministry of Science, Innovation and Universities MA is supported by a postdoctoral fellowship 'Juan de la Ciervaincorporacion' from the Ministry of Science, Innovation and Universities. The funding sources had no involvement in study design; in the collection, analysis and interpretation of data; in the writing of the report; and in the decision to submit the article for publication.
The data that support the findings of this study are available from the corresponding author upon reasonable request.