Using lymphatic pumping techniques to determine the key mediators of Inflammatory Bowel Disease

Barcroft M, Barcroft Z, Atkinson E, Berglind J
Edward Via College of Osteopathic Medicine- Carolinas Campus, Spartanburg, SC, United states

Purpose & Aims
Inflammatory bowel disease (IBD), comprised of ulcerative colitis (UC) and crohn’s disease (CD), is known to have a multifactorial process involving the immune, lymphatic, and gastrointestinal systems. The peak incidence of diagnosis for IBD is between the ages of 15 to 30, with significant complications specific to the pediatric population including growth failure and puberty delay. IBD is known to have both genetic and environmental causes; however, the exact pathophysiology is unknown. While there are various treatment modalities available including immunosuppression, ultimately one third of UC patients and over half of CD patients end up requiring surgery. For this reason, there is a significant need to improve medical therapies for IBD.
In the gut, the lymphatics have shown to control tissue edema, leukocyte trafficking, and chemokine clearance. Therefore, lymph stasis is thought to be a major contributing factor to the disruption of the normal immune and wound healing response to the tissue damage seen in IBD. If lymph flow can be restored, then lymphatic remodeling and the body’s natural self-healing mechanisms can overcome the innate immune response. We hypothesize that osteopathic manipulative treatment (OMT), specifically lymphatic pumping techniques (LPT), can be used to increase immune cell mobilization, antigen and cytokine clearance, while also promoting proper wound healing in an IBD mice model.
In this study, we used C57BL6 mice treated with a 3% solution of dextran sulfate sodium (DSS). This method is a well-established model showing similar clinical features to IBD, with minimal risk of mortality. The mice were divided into four groups: control with normal water, control with 3% DSS, anesthesia with 3% DSS, and anesthesia with LPT and 3% DSS. All groups received the respective fluid intake for four days, with euthanasia and colon collection following on the fifth day. For the groups requiring treatment, the anesthesia and LPT were performed 24 hours after the initial start time, once daily for the first four days. The LPT group received daily treatment by contacting the abdomen and pumping at a rate of 1/sec. for 4 minutes. Following euthanasia and colon collection, all samples were homogenized or fixed in formalin. The homogenized colons were run using ELISA for multiple inflammatory markers including, but not limited to, IL-1𝝱, IL-2, IL-5, IL- 12 p70, IL-17 A Homodimer, IL-22, IL-23, and TGF-𝝱.
While full data analysis of this larger study has not been completed, preliminary studies have shown significant differences between the OMT groups and the control groups. The mice that received OMT showed less weight loss and a decreased innate response shown by analyzing TNF-𝛂 and IFN-𝛄 levels. The broader analysis of inflammatory markers, along with a larger sample size will allow us to further explore the dynamics between the lymphatic system and inflammatory bowel disease.