Quantitative assessment of intestinal stiffness and associations with fibrosis in human inflammatory bowel disease.

Daniel C Stewart,Xinyue Liu,Chelsey S Simmons,Andria L Doty,Sarah C Glover,Atif Iqbal,Cooper Rickerson,David Mkoji,Dalton Berrie,Jian Li,Sanda Tan,Shree Ram Singh

doi:10.1371/journal.pone.0200377

Abstract

Inflammatory bowel disease (IBD) continues to increase in prevalence in industrialized countries. Major complications of IBD include formation of fibrotic strictures, fistulas, reduced absorptive function, cancer risk, and the need for surgery. In other chronic gastrointestinal disease models, stiffness has been shown to precede fibrosis; therefore, stiffness may be a reasonable indicator of progression toward stricture formation in IBD patients. Herein, we seek to quantify tissue stiffness and characterize fibrosis in patients with IBD and to compare mechanical properties of unaffected human tissue to common animal species used for IBD studies. Inflamed and unaffected tissue from IBD patients and unaffected tissue from mice, pigs, and cows were indented using a custom device to determine the effective stiffness. Histology was performed on matched tissues, and total RNA was isolated from IBD tissue samples and used for gene expression analysis of pro-fibrotic genes. We observed an increase in the effective stiffness (steady-state modulus, SSM) (p < 0.0001) and increased expression of the collagen type I gene (COL1A1, p = 0.01) in inflamed tissue compared to unaffected areas in our IBD patient cohort. We also found that increased staining of collagen fibers in submucosa positively correlated with SSM (p = 0.093). We determined that unaffected animal bowel stiffness is significantly greater than similar human tissues, suggesting additional limitations on animal models for translational investigations regarding stiffness-related hypotheses. Taken together, our data support development of tools for evaluation of bowel stiffness in IBD patients for prognostic applications that may enable more accurate prediction of those who will develop fibrosis and more precise prescription of aggressive therapies.

Highlights

Inflammatory bowel diseases (IBD) are chronic autoinflammatory disorders that can result in Crohn’s disease (CD) or ulcerative colitis (UC)
Sectioned tissue was placed in a cold media solution consisting of Dulbecco’s Modified Eagle Medium (DMEM) and 10% Fetal Bovine Serum (FBS) for subsequent analysis, and samples for mechanical characterization were stored on ice and tested within 4 hours of isolation, similar to previous work characterizing gastrointestinal tissue [30]
Our direct characterization has established that inflamed regions of human bowel are significantly stiffer than uninflamed regions and has confirmed the general trends observed previously via compression [48] and ultrasound elastography [15,49,50,51,52]

Summary

Introduction

Inflammatory bowel diseases (IBD) are chronic autoinflammatory disorders that can result in Crohn’s disease (CD) or ulcerative colitis (UC). CD is associated with transmural inflammation in regions throughout the gastrointestinal tract, and approximately 40% to 71% of all CD patients will develop complications requiring surgical resection of the affected bowel segments within 10 years of diagnosis [2,3,4,5]. There is a growing interest in developing non-invasive methods to evaluate fibrosis in all IBD patients because new therapies that target fibrosis could help to avoid or delay surgery [10,11,12,13]. Non-invasive procedures such as MRI enterography are beneficial for diagnosis, but only advanced fibrosis can be detected [3,14,15]. Stiffness monitoring could be used as a non-invasive method for early detection of disease acceleration.

Methods

Results

Conclusion