Abstract
Foreign bodies such as fibers of a surgical mesh induce a typical reaction with an inflammatory infiltrate that forms a surrounding granuloma. This infiltrate is dominated by macrophages, lymphocytes, and neutrophils, whereas its extent of collaboration is widely unknown. In this study, we analyzed 12 samples of surgical meshes explanted from humans by multiplex analyses with three different 5-marker panels – 1. macrophage panel: CD68, CD86, CD105, CD163, and CD206; 2. lymphocyte panel: CD3, CD4, CD8, CD20, and CD68; and 3. neutrophil panel: CD15, histone, MPO, NE, and CD68. Measurement of fluorescence intensity within nuclear masks resulting from DAPI nuclear staining allows exact quantification of cells considered “positive” at a user-defined mean intensity threshold of > 100. Obviously, however, there is no natural threshold as a biological criterion for an intensity that separates “positive” stained cells from unstained cells (“negative”). Multiplex staining of 5 markers always reveals a high rate of coexpression for almost all of the 25 possible marker combinations (= 32 combinations, when using 5 markers simultaneously). The present staining results demonstrate that various morphological and functional subtypes of macrophages, lymphocytes, and neutrophils are abundant in the foreign body granuloma (FBG), which were investigated by regions of interest (ROI) with an area of 1 mm2. The widespread coexpression of two or more markers underscores the complex collaboration network of the inflammatory infiltrate. The ability to combine spatial distribution with exact numerical analysis may offer new perspectives for our understanding of the complex interactions in this multidimensional process.
Highlights
The repair of hernias as defects of the abdominal wall with possible protrusion of intestine is the most frequent procedure of visceral surgery
These porous devices elicit a foreign body reaction that culminates in the formation of a foreign body granuloma (FBG), consisting of an inflammatory infiltrate surrounded by a fibrotic capsule
The cells of the inflammatory infiltrate around mesh fibers showed a variable signature of macrophage, lymphocyte, and neutrophil markers in multiplex immunofluorescence microscopy
Summary
The repair of hernias as defects of the abdominal wall with possible protrusion of intestine is the most frequent procedure of visceral surgery. The closure of the hernia orifice turned from simple suture to extended reinforcement with non-absorbable textile structures, the so-called meshes. These porous devices elicit a foreign body reaction that culminates in the formation of a foreign body granuloma (FBG), consisting of an inflammatory infiltrate surrounded by a fibrotic capsule. Measurement of the mean intensities in the area of a nucleus offers the possibility of precise quantification of the number of “positive” cells and characterization of the local cellular response to the foreign body
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