Immunohistochemical Analysis of Matrix Proteolytic Enzymes in the Periprosthetic Tissue in the Patients with Loosening Prostheses.

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Abstract
Regional periprosthetic bone resorption plays an important role of prosthesis loosening. In order to study the possible mechanisms of loosening, we investigated the presence of matrix proteolytic enzymes in the periprosthetic tissue by immunohistochemical technique in 72 patients undergoing revision operation of loosened joint prosthesis, including 22 males and 50 females and aged from 19 to 88 years (mean, 61.7 years). Thirty-nine patients had a loosened hip prosthesis (18 males and 21 females) whereas 33 patients had a loosened knee prosthesis (4 males and 29 females). Tissue specimens collected during revision surgery underwent thin slide sections and H & E staining, and were observed under light microscopy and polarized-light microscopy. The results showed many macrophages, histiocytes, fibroblasts, as well as many phagocytosed metal debris and polyethylene debris in the periprosthetic tissues, suggesting an active bone resorption. Furthermore, we used immunohistochemical techniques to detect the distribution of matrix proteolytic enzymes in periprosthetic tissue, including lysosome enzymes (cathepsin B, cathepsin D and cathepsin G), and matrix metalloproteinase (MMPs, MMP-1, MMP-2, MMP-3). The immunostaining were classified as strong positivity, > 70% positive cells; moderate positivity, 20-70% positive cells; weak/negative, < 20% positive cells. The results showed that cathepsin B, cathepsin D and cathepsin G were found in most fibroblasts and macrophage-like cells, including multinuclear giant cells and epithelioid cells. MMPs were found in most fibroblasts and macrophage-like cells, as well as a scant amount in the extracellular matrix. These enzymes were also found in or around blood vessels, the endothelial cells in the richly vascularized tissue. All negative controls showed no staining. The results of immunoreactive staining ranged from 61.1% to 68.1% of strong to moderate positivity. Since these enzymes were related to the degradation of matrix protein, they may be related to the periprosthetic bone resorption. The further clinical significance needs further investigation.