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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