J Pathol Transl Med.  2022 Sep;56(5):239-248. 10.4132/jptm.2022.05.18.

Inflammation and tissue remodeling contribute to fibrogenesis in stricturing Crohn’s disease: image processing and analysis study

• Arslan ME
• Brar R
• Goetz L
• Karamchandani D
• Mikula MW
• Hodge K
• Li H
• Ahn S
• Lee H

• KMID: 2533700
• DOI: http://doi.org/10.4132/jptm.2022.05.18

Abstract

Background
Inflammation and structural remodeling may contribute to fibrogenesis in Crohn’s disease (CD). We quantified the immunoexpression of calretinin, CD34, and calprotectin as a surrogate for mucosal innervation, telocytes (interstitial cells playing a role in networking), and inflammation, respectively, and correlated them with bowel alterations in stricturing CD.
Methods
Primary resection specimens for ileal CD (n = 44, 31 stricturing CD, 13 inflammatory CD) were identified. Left-sided ulcerative colitis and trauma cases were used as controls. Proximal and distal margin and middle (diseased) sections were stained for calretinin, CD34, and calprotectin. Microscopic images were captured from the mucosa (calretinin), submucosa (calprotectin), and myenteric plexus (CD34), and the immunostaining was quantified using image processing and analysis. Bowel thickness at the corresponding sections were measured and correlated with the amount of immunoexpression.
Results
A total of 2,037 images were analyzed. In stricturing CD, submucosal alteration/thickening at the stricture site correlated with calprotectin staining and inversely correlated with calretinin staining at the proximal margin. Muscularis propria alteration/thickening at the stricture site correlated with mucosal calretinin staining at the proximal margin. Submucosal alteration/thickening at the proximal margin correlated with calretinin and CD34 staining at the proximal margin and inversely correlated with CD34 staining at the stricture site. Calretinin immunostaining at the distal margin was significantly higher in stricturing CD than the controls.
Conclusions
Inflammation and tissue remodeling appear to contribute to fibrogenesis in stricturing CD. Increased mucosal calretinin immunostaining distal to the diseased segment could be helpful in diagnosing CD in the right clinical context.

Keyword

Crohn’s disease; Stricture; Fibrosis; Telocyte; Calretinin; Calprotectin

Figure

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  Fig. 1. Diagram demonstrating sections and immunohistochemistry.

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  Fig. 2. An example of calretinin immunohistochemistry and image processing and analysis. (A) Proximal margin mucosa (calretinin immunostain). (B) Spaces and crypts are segmented (black). (C, D) Brown-colored pixel within the mucosa.

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  Fig. 3. An example of CD34 immunohistochemistry and image processing and analysis. (A) Myenteric plexus layer (CD34 immunostain). (B) Spaces are segmented (black). (C, D) Brown-colored pixel in the remaining background tissue.

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  Fig. 4. An example of calprotectin immunohistochemistry and image processing and analysis. (A) Submucosa (calprotectin immunostain). (B) Artifactual spaces and vascular channels are segmented (black). (C, D) Brown-colored pixel in the remaining background tissue.

Reference

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