Nuovi Elementi di OsteoImmunologia nell'Osteolisi Periprotesica
AOU Molinette di Torino SOC OrtopediaDir. ff dott Nicola Ruggieri
Scuola di Dottorato in Biotecnologie Università di Genova
Corso di Laurea in Scienze Motorie Università di Genova
Prof. Luigi MolfettaPresidente CdL
Prof. Rodolfo QuartoProf. Ordinario Biotecnologie
Dr. Davide CaldoDirigente Medico OrtopediaDottorando III aa
THA: Best Surgical Procedure in all medical assets Aseptic Looseningè P. Osteolysisè Wear Debris
80% A.L.
Background
Holt G, Murnaghan C, et al. (2007). The biology of aseptic osteolysis. Clin Orthop Relat Res 460: 240-52.
Maitra R, Clement CC, et al. Endosomal damage and TLR2 mediated inflammasome activation by alkane particles in the generation of aseptic osteolysis. Mol Immunol. 2009 Dec;47(2-3):175-84. Epub 2009 Oct 4.
+
TNF alpha IL6
+RANKL
Sabokbar, A., I. Itonaga, et al. (2005). "Arthroplasty membrane-derived fibroblasts directly induce osteoclast formation and osteolysis in aseptic loosening." J Orthop Res 23(3): 511-9.
T cell in Aseptic LooseningT cells in periprosthetic tissue = 1-16%Low expression of activation markers and citokynes (CD25, IL1, IL2, IL6, TNFalpha)
T cells do NOT play a pivotal role aseptic loosening
Baldwin, L., B. Flanagan, et al. (2002). "A study of tissue interface membranes from revision accord knee arthroplasty: the role of T lymphocytes." Biomaterials 23(14): 3007-14
Li, T., S. Santavirta, et al. (2001). "No lymphokines in T-cells around loosened hip prostheses." Acta Orthop Scand 72(3): 241-7.
Circulating committed T cells in osteolysis of various
etiologyRoato, I., M. Grano, et al. (2005). "Mechanisms of spontaneous
osteoclastogenesis in cancer with bone involvement." Faseb J 19(2):
228-30
OC negative feedback on T cell, by priming CD8+foxp3+
suppressorKiesel, J. R., Z. S. Buchwald, et al.
(2009). "Cross-presentation by osteoclasts induces FoxP3 in CD8+ T
cells." J Immunol 182(9): 5477-87-30
OSTEOIMMUNOLOGY
Obiettivi:
• dimostrare la presenza di Tcell circolanti committed to osteoclastogenesis
• dimostrare la presenza nei tessuti periprotesici di linfociti T inibitori col fenotipo tipico conseguente al priming dagli OC
HYPOTHESIS: (Nelle fasi precoci dellʼ) osteolisi cʼè un ruolo pro-osteoclastogenetico dei linfociti T, in presenza di numero critico di OC nei tessuti periprotesici (condizione degli studi !!!) prevale il feedback negativo da essi generato
Methods45 patients in 3 groups:- 15 PO- 15 stable prosthesisAge and sex matched
-15 healthy controls
Exclusion criteria = any condition affecting bone metabolism
Peripheral BloodIn vitro spontaneous
osteoclastogenesis: trinucl TRAP+ d13°/15°
resorption test T depletion RANKFc
Cytokine: Elisa for IL-7
Immunofenotipi T cell: FACS for CD3/4/8/69/25
Methods
MethodsPeriprosthetic TissuesInteractions between OC
and T cells H&E and IF
Immunophenotypes of T cells
IHC and FACS
Results
- incremento assoluto numero di T cell medio, sia CD4 che CD8 (n.s.)- assenza di CD8+CD25+ circolanti nel sangue dei pazienti in cui avviene socg
Results
IHC
Results
IHC showed expression of
CD4, CD8, CD25 and Foxp3 on
periprosthetic membranes
ResultsFACS
• CD3 = 1%• CD4+ CD25- Foxp3- • CD8+ CD25+ Foxp3+ =• CD8+/CD25+/FoxP3+ also positive for TGF-β
1.4%
TNF alpha IL6
RANKL
/
• Our data suggest T cells may have a role in early Periprosthetic Osteolysis
• Reaching a critical OC number (advanced stage, radiographic +) CD8+foxp3+ T Reg turn off lymphocyte activity
• Conferme sperimentali necessarie
Conclusions
• Da interpretare il quadro immunofenotipico periferico
• If data will be confirmed, therapeutic implications need to be investigated (routinary prophilaxis with bisphosphonate in THA???)
Conclusions
• Monitoraggio in vivo, con lab-on-a-chip• Osteoimmunogenetica: variabilità delle
molecole coinvolte e predisposizione al fallimento precoce/ massivo
Sviluppi Futuri
Thanks for your attention