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A STUDY OF THE MECHANISM OF PERIPROSTHETIC OSTEOLYSIS USING SURGICAL SPECIMENS: HISTOLOGICAL AND IMMUNOHISTOCHEMICAL STUDY



Abstract

Periprosthetic osteolysis has attracted attention as a cause of loosening after arthroplasty. The aim of the present study was to examine inflammatory cell localization and the occurrence of apoptosis in granulation tissue from patients who required revision arthroplasty due to loosening caused by osteolysis.

7 patients were studied comprising 3 patients who underwent FHR and 4 patients who underwent THR. Their mean age at the time of surgery was 63.6 years. The mean period from their previous operation to revision was 8.8 years.

Granulation tissue was collected from around the loosened implant fixed in 4% paraformaldehyde and embedded in paraffin. Sections were cut and were first stained with hematoxylin and eosin. Next, immunohistochemical studies were performed using the avidin-biotin complex method. CD45 was used as the primary antibody to detect T cells, and CD68 was used to detect macrophage-like cells. The activity of the macrophage-like cells was assessed with anti-I-NOS and anti-MMP-9.

Apoptosis was investigated using anti-single-stranded DNA (ssDNA). Using another granulation tissue was stored at −80%C, DNA was extracted, and the presence of DNA fragmentation was detected by agarose gel electrophoresis.

Vascularization and infiltration by a large number of inflammatory cells were seen along with large multinucleated osteoclas-like cells. Immunohistochemical studies revealed CD45-positive cells primarily around the blood vessels. The CD68-positive cells were mainly multinucleated cells. The multinucleated cells were i-NOS-positive in 4 patients, and were MMP-9-positive in 5 patients.

The nuclei of many of the multinucleated cells were positive for ssDNA. Agarose gel electrophoresis of DNA showed a marked ladder pattern at the 170 base pair region. This finding indicated DNA fragmentation or apoptosis.

Apoptotic cells were seen in granulation tissue harvested from around loosened implants suggesting that apoptosis may play a role in the pathophysiology of osteolysis.

The abstracts were prepared by Mr Simon Donell. Correspondence should be addressed to him at the Department of Orthopaedics, Norfolk & Norwich Hospital, Level 4, Centre Block, Colney Lane, Norwich NR4 7UY, United Kingdom.