IBD

Inflammation Assays

Inflammatory Bowel Disease (IBD)

IBD is a term used to describe disorders that involve chronic/recurrent inflammation of the colon and small intestine.

Ulcerative colitis is mainly localised into the colon (large intestine), while Crohn’s disease can affect any part of the digestive system. In both cases, symptoms may range from mild to severe, with periods of active illness followed by temporary remission. There is currently no cure for IBD, and medications tend to reduce the symptoms preventing severe complications, which include colon cancer.

Causes:

Defects in the intestinal barrier
Impaired immune function
Genetic predispositions (e.g. NOD2 mutation)
Environmental factors

Characteristics:

Increased proinflammatory immune response
Increased permeability of the intestinal epithelial barrier
Increased mucus production
Rearrangement of tight junction proteins

In vitro models –

Relevant cell types:

Caco-2 cells – Human Colorectal adenocarcinoma epithelial-like cell line
InEpC – Primary Human Intestinal Epithelial Cells
Immune cells – Freshly isolated from peripheral blood of healthy/diseased volunteers

Caco-2 Model for IBD: Inflammatory Mediators

Caco-2 cells were differentiated for 21 days and subsequently treated with Dexamethasone, followed by stimulation with LPS+ IL1beta cocktail. mRNA expression in cell lysates was quantified using the Luminex Quantigene™ Assay. The supernatants were analysed for inflammatory mediators using a Luminex Multiplex Assay (**p<0.01; ***p<0.001; ****p<0.0001; n=3 mean±SEM).

Pro-inflammatory marker suppression by Sulfasalazine and 5-Aminosalicylic Acid

Significantly increased levels of pro-inflammatory markers were observed after stimulation of the differentiated Caco-2 monolayers with IL-1β and LPS when compared to unstimulated control. The levels of these markers were suppressed when the Caco-2 monolayers were pre-treated with the reference compounds. Data was analyzed using Ordinary One-way ANOVA followed by Dunnett’s multiple comparisons test [95% CI of diff.] (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ns= not significant). All the conditions were compared to the stimulated control for each of the markers and showed significant differences.

Note: Unstimulated = Co-culture (Caco-2 +THP-1), Stimulated= Co-culture (Caco-2 +THP-1) + LPS + IL-1β.

Lactate Dehydrogenase (LDH) release by Caco-2 cells

Data was analyzed using Ordinary One-way ANOVA followed by Dunnett’s multiple comparisons test [95% CI of diff.] (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ns= not significant). The LDH positive control, provided with the assay, generated significant OD 450 values (p<0.0001) compared to the unstimulated control. Both Sulfasalazine and 5-Aminosalicylic Acid showed no significant LDH release when compared to the unstimulated control.

FAQ's

What are the big challenges in IBD?

IBD is a highly complex disease involving dysregulated T cell responses, epithelial barrier dysfunction, microbiome interactions, and chronic inflammation. Because of this complexity, in vitro models need to capture key aspects of human intestinal biology to generate meaningful and translational data.

Importance of human-relevant in vitro models ?

Human-relevant in vitro models can provide mechanistic insights that are difficult to obtain using conventional animal models. They allow researchers to study epithelial integrity, cytokine responses, and immune signalling under controlled laboratory conditions.

What are some co-culture systems for IBD research?

Our research platforms support studies in multiple respiraCo-culture systems combining epithelial and immune cells are commonly used to investigate inflammatory responses, epithelial barrier disruption, and mechanisms of action of therapeutic compounds. These models help better represent the cellular interactions seen in the intestinal environmenttory diseases, including asthma, airway inflammation, pulmonary fibrosis, COPD, viral infection and allergic respiratory conditions.

Commonly assessed endpoints in inflammatory bowel disease models?

Common endpoints include cytokine quantification, TEER and permeability assays to assess barrier integrity, gene expression profiling, and analysis of inflammatory biomarkers associated with intestinal dysfunction

The Potential of In Vitro Models of Inflammatory Bowel Disease in the Discovery of New Drugs?

In vitro IBD models provide reproducible platforms to evaluate anti-inflammatory and barrier-protective effects of new therapies. They are widely used to support target validation, compound screening, and mechanistic studies during drug development

Request a consultation with Cellomatics Biosciences today

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To request a consultation where we can discuss your exact requirements, please contact Cellomatics Biosciences.