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Immunology and Infection

Systematic Scoring Analysis for Intestinal Inflammation in a Murine Dextran Sodium Sulfate-Induced Colitis Model

Published: February 14th, 2021



1Department of Pathology, University of Michigan, 2Department of Surgery, Klinikum Rechts der Isar, Technische Universität München, 3Wallenberg Centre for Molecular Medicine, Linköping University, 4Department of Biomedical and Clinical Sciences (BKV), Linköping University

Systematic scoring of intestinal inflammation using a free computer-assisted system is a powerful tool to quantitatively compare histopathological changes in colitis models characterized by the presence of ulcers and inflammatory changes. Histological colitis score evaluation strengthens clinical observations and facilitates data interpretation.

Murine colitis models are tools that are extensively employed in studies focused on understanding the pathobiology of inflammatory intestinal disorders. However, robust standards for objective and reproducible quantification of disease severity remain to be defined. Most colitis analysis methods rely on limited histological scoring of small segments of intestine, leading to partial or biased analyses. Here, we combine high-resolution image acquisition and longitudinal analysis of the entire colon to quantify intestinal injury and ulceration in the dextran sodium sulfate (DSS) induced model of murine colitis. This protocol allows for the generation of objective and reproducible results without extensive user training. Here, we provide comprehensive details on sample preparation and image analysis using examples of data from DSS induced colitis. This method can be easily adapted to other models of murine colitis that have significant inflammation associated with mucosal injury. We demonstrate that the fraction of inflamed/injured and eroded/ulcerated mucosa relative to the complete length of the colon closely parallels clinical findings such as weight loss amid DSS-induced disease progression. This histological protocol provides a reliable time and cost-effective aid to standardize analyses of disease activity in an unbiased way in DSS colitis experiments.

The gastrointestinal epithelial barrier plays a pivotal role in separating luminal antigens and pathogens from underlying tissue compartments1. Epithelial injury and mucosal wounds seen in pathologic conditions such as inflammatory bowel disease (IBD), ischemia, or surgical injury are associated with clinical symptoms that include diarrhea, weight loss, blood in stool, and abdominal pain. In response to injury, epithelial cells migrate and proliferate to re-epithelialize and repair mucosal barrier defects. Resolution of inflammation and restitution of mucosal integrity are crucial to re-establishing intestinal mucosal homeostasis and function

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All animal experiments described were approved by the University of Michigan's Committee on the Use and Care of Animals.

1. Tissue harvest

  1. Euthanize mice humanely using isoflurane anesthesia followed by cervical dislocation, in accordance with approved protocols. For all animal experiments, approval was obtained by a certified review board in accordance with the National and Institutional guidelines for animal handling.
  2. Place the mouse on a dissecting pad in a supine .......

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To illustrate the reliability of this Histological Colitis Score Analysis in the context of mucosal damage after DSS challenge and subsequent recovery from colitis, we administered 2.5% DSS in the drinking water of eight 10 weeks old male C57BL6 wild type mice for 5 days followed by a recovery period with regular water for 5 days. There was no change in body weight during the acute administration of DSS, from day 0 to 5 (Figure 4A). Body weight dramatically d.......

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Our Histological Colitis Score system constitutes a reliable tool to quantify tissue inflammation and damage in the intestine. This approach provides an improved understanding of the histopathological state of the whole organ without the bias of selecting small areas or incomplete sections. Among the critical steps to successfully execute this protocol are proper preparation of Swiss rolls that allow for analysis of at least 90% of the length of the colon; parallel orientation during paraffin embedding and sectioning to .......

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The authors wish to acknowledge support from NIH funding DK055679, DK089763, DK079392, DK061739, DK072564 and the University of Michigan Pathology Slide Scanning Services.


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Name Company Catalog Number Comments
Aperio AT2 – High Volume, Digital Whole Slide Scanning Leica Biosystems Aperio AT2
Absorbent Underpads with waterproof moisture barrier VWR International 56616-032
American Line 66-0089 Single Edge Blade, 100 per pkg GT Midwest TL5837
BD Luer-Lok Disposable Syringes without Needles Fisher scientific 14-823-2A
Bonn Strabismus Scissors - ToughCut Fine Science tools 11103-09
Bonn Strabismus Scissors - ToughCut Fine Science tools 14084-09
Dumont #5 Forceps Fine Science tools 11251-20
Formalin solution, neutral buffered, 10% Sigma HT501640-19L
HistoPrep 70% Ea Fisherbran 70% denatured ethyl alcohol
ImageScope Aperio Version
LeakBuster Specimen Containers: Sterile Starplex Scientific B120210
Phosphate-Buffere Saline, without calcium & magnesium Corning 21-040-CV
Plastic Feeding tubes, 20 GA x 30 mm Instech FTP2030
PrecisionGlide Needle, Size: 20 G x 1 1/2 in BD (Becton, Dickinson and Company) 305176
PrecisionGlide Needle, Size: 27 G x 1/2 in BD (Becton, Dickinson and Company) 305109
Syringe, 10 ml BD (Becton, Dickinson and Company) 302995
Unisette Tissue Cassettes Simport M505-2

  1. Kagnoff, M. F. The intestinal epithelium is an integral component of a communications network. Journal of Clinical Investigation. 124 (7), 2841-2843 (2014).
  2. Laukoetter, M. G., Nava, P., Nusrat, A. Role of the intestinal barrier in inflammatory bowel disease. World Journal of Gastroenterology. 14 (3), 401-407 (2008).
  3. Baumgart, D. C., Sandborn, W. J. Crohn's disease. Lancet. 380 (9853), 1590-1605 (2012).
  4. Ordas, I., Eckmann, L., Talamini, M., Baumgart, D. C., Sandborn, W. J. Ulcerative colitis. Lancet. 380 (9853), 1606-1619 (2012).
  5. Vowinkel, T., Kalogeris, T. J., Mori, M., Krieglstein, C. F., Granger, D. N. Impact of dextran sulfate sodium load on the severity of inflammation in experimental colitis. Digestive Diseases and Sciences. 49 (4), 556-564 (2004).
  6. Kitajima, S., Takuma, S., Morimoto, M. Histological analysis of murine colitis induced by dextran sulfate sodium of different molecular weights. Experimental Animals. 49 (1), 9-15 (2000).
  7. Mahler, M., et al. Differential susceptibility of inbred mouse strains to dextran sulfate sodium-induced colitis. American Journal of Physiology. 274 (3), 544-551 (1998).
  8. Wirtz, S., Neufert, C., Weigmann, B., Neurath, M. F. Chemically induced mouse models of intestinal inflammation. Nature Protocols. 2 (3), 541-546 (2007).
  9. Kozlowski, C., et al. An entirely automated method to score DSS-induced colitis in mice by digital image analysis of pathology slides. Disease Models & Mechanisms. 6 (3), 855-865 (2013).
  10. Perse, M., Cerar, A. Dextran sodium sulphate colitis mouse model: traps and tricks. Journal of Biomedicine and Biotechnology. 2012, 718617 (2012).
  11. Mizoguchi, A. Animal models of inflammatory bowel disease. Progress in Molecular Biology and Translational Science. 105, 263-320 (2012).
  12. Flemming, S., et al. Desmocollin-2 promotes intestinal mucosal repair by controlling integrin-dependent cell adhesion and migration. Molecular Biology of the Cell. 31 (6), 407-418 (2020).
  13. Luna, L. G. . Armed Forces Institute of Pathology (U.S) & Armed Forces Institute of Pathology (U.S.). Manual of Histologic Staining Methods of the Armed Forces Institute of Pathology. 3d edn. , (1968).
  14. Kelm, M., et al. Targeting epithelium-expressed sialyl Lewis glycans improves colonic mucosal wound healing and protects against colitis. The Journal of Clinical Investigation Insight. 5 (12), (2020).
  15. Reed, M., et al. Epithelial CD47 is critical for mucosal repair in the murine intestine in vivo. Nature Communications. 10 (1), 5004 (2019).
  16. Laukoetter, M. G., et al. JAM-A regulates permeability and inflammation in the intestine in vivo. Journal of Experimental Medicine. 204 (13), 3067-3076 (2007).
  17. Khounlotham, M., et al. Compromised intestinal epithelial barrier induces adaptive immune compensation that protects from colitis. Immunity. 37 (3), 563-573 (2012).

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