PhD Scientific Days 2018

Budapest, April 19–20, 2018

Developing a novel diagnostic method in periprosthetic joint infection by detecting neutrophil derived microvesicles in synovial fluid

Sallai, Imre

1. Dr. Sallai Imre; Semmelweis University Department of Orthopaedics, Budapest
2. Dr. Marton Nikoletta, Semmelweis University Department of Genetics, Cell- and Immunobiology, Budapest
3. Gál Domokos, Semmelweis University Department of Orthopaedics, Budapest
4. Kittel Ágnes, Hungarian Academy of Sciences, Budapest
5. Dr. Nagy György, Semmelweis University Department of Genetics, Cell- and Immunobiology, Budapest
6. Dr. Buzás Edit, Semmelweis University Department of Genetics, Cell- and Immunobiology, Budapest
7. Dr. Kristóf Katalin, Semmelweis University Clinical Microbiological Diagnostic Laboratory, Budapest
8. Dr. Komlósi Zsolt, Semmelweis University Department of Pulmonology
9. Dr. Daniel Kendoff, Helios Klinikum Berlin-Buch, Berlin
10. Dr. Zahár Ákos, HELIOS ENDO-Klinik, Hamburg
11. Dr. Skaliczki Gábor; Semmelweis University Department of Orthopaedics, Budapest

Language of the presentation

Hungarian

Text of the abstract

Introduction: joint surface replacement is one of the most successfull procedures in orthopedic surgery. Bacterial infections are the most devastating complications of this type of operations. Despite the development of the diagnostic tools, the management of the septic implant is a great challenge for the orthopedic surgeon. The rates of septic complications are between 0,5-2,5% after a primary procedure all over the word. The role of polymorphonuclear neutrofils (PNMs) during bacterial infections is well known but in the last decade it was observed that after bacterial opsonisation they are increasing the release of microvesicles against germs.
Aims: the aim of our research was to investigate the role of microvesicles released by PMNs in periprosthetic joint infections. Considering that the properties of PMNs derived microvesicles were not investigated previously from synovial fluid around orthopedic implants, we developed an isolation protocol applicable for septic and sterile sample processing.
Method: samples were taken during revision surgeries previously proved to be septic and sterile loosening. Four septic and four sterile samples were collected and synovial fluids were prepared following our own isolation protocol. PMNs microvesicles were characterized by flow cytometry using the specific CD11b, CD66b, CD177 and Annexin V. antibodies. For the reverse proof of vesicle origin Triton X-100 detergent was used. Some samples were investigated by electron microscope to visualize the vesicles and their properties.
Results: many literature recommendations were used to develop, improve and verify a standard microvesicle isolation protocol which is applicable for synovial fluid from both septic and sterile loosening. Quantitative and qualitative differences were proven by flow cytometric and electron microscopic investigations between the septic and sterile loosening samples.
Conclusion: after bacterial exposure the number and properties of microvesicles released by PMNs changed providing promising results in the development of diagnostics of orthopedic implant associated infections.

Data of the presenter

Doctoral School: Clinical Medicine
Program: Physiology and Pathology of the Musculoskeletal System
Supervisor: Gábor Skaliczki
E-mail address: sallai01imre@gmail.com