Transplantation of human intestinal organoids into the mouse mesentery: A more physiologic and anatomic engraftment site

International audienc

Knowledge, attitude and awareness among diabetic vs. non-diabetic patients about the association between diabetes and oral health

Background: Global prevalence of diabetes is rapidly increasing with an estimate to affect 593 million worldwide by 2035. Current evidence clearly states an association between oral diseases and diabetes mellitus with manifestations like periodontitis, peri-implantitis, xerostomia, etc. Despite this obvious link, knowledge, awareness and attitude of general population towards this are not fully understood. Aims: To assess public knowledge and awareness on association between diabetes and oral health and assess their attitude towards oral hygiene care and maintenance. Methods and Materials: A three-part structured questionnaire was developed with multiple choice questions and circulated among patients visiting a private dental college. A total of 502 questionnaires were evaluated, and results were statistically analysed. Statistical Analysis Used: Student-t test and Chi-square test using SPSS software. Results: Majority of the participants were male and lived in urban area. 41.8% respondents had diabetes, out of which 86.7% had no awareness of the type. A significant number of non-diabetic individuals had awareness of excessive sugar intake as a cause of diabetes. Greater number of diabetic patients (96.7%) reported dry mouth, whereas only 53.3% had periodontal complications. 90% participants use tooth brush as oral hygiene aid. Only 10.6% participants follow regular dental visits. Majority of subjects (60.6%) listed their friends and family as major source of information. Conclusion: More people are aware of systemic complications of diabetes as compared to oral problems. A better interdisciplinary relationship is required among dentists and physicians to improve knowledge and awareness of general population

Evaluation of transplantation sites for human intestinal organoids.

Our group has developed two transplantation models for the engraftment of Human Intestinal Organoids (HIOs): the renal subcapsular space (RSS) and the mesentery each with specific benefits for study. While engraftment at both sites generates laminated intestinal structures, a direct comparison between models has not yet been performed. Embryonic stem cells were differentiated into HIOs, as previously described. HIOs from the same batch were transplanted on the same day into either the RSS or mesentery. 10 weeks were allowed for engraftment and differentiation, at which time they were harvested and assessed. Metrics for comparison included: mortality, engraftment rate, gross size, number and grade of lumens, and expression of markers specific to epithelial differentiation, mesenchymal differentiation, and carbohydrate metabolism. Mortality was significantly increased when undergoing mesentery transplantation, however engraftment was significantly higher. Graft sizes were similar between groups. Morphometric parameters were similar between groups, however m-tHIOs presented with significantly fewer lumens than k-tHIO. Transcript and protein level expression of markers specific to epithelial differentiation, mesenchymal differentiation, and carbohydrate metabolism were similar between groups. Transplantation into both sites yields viable tissue of similar quality based on our assessments with enhanced engraftment and a dominant lumen for uniform study benefiting the mesenteric site and survival benefiting RSS

WNT2B Deficiency Causes Increased Susceptibility to Colitis in Mice and Impairs Intestinal Epithelial Development in Humans.

WNT2B is a canonical Wnt ligand previously thought to be fully redundant with other Wnts in the intestinal epithelium. However, humans with WNT2B deficiency have severe intestinal disease, highlighting a critical role for WNT2B. We sought to understand how WNT2B contributes to intestinal homeostasis. We investigated the intestinal health of Wnt2b knock out (KO) mice. We assessed the impact of inflammatory challenge to the small intestine, using anti-CD3χ antibody, and to the colon, using dextran sodium sulfate (DSS). In addition, we generated human intestinal organoids (HIOs) from WNT2B-deficient human iPSCs for transcriptional and histological analyses. Mice with WNT2B deficiency had significantly decreased Lgr5 expression in the small intestine and profoundly decreased expression in the colon, but normal baseline histology. The small intestinal response to anti-CD3χ antibody was similar in Wnt2b KO and wild type (WT) mice. In contrast, the colonic response to DSS in Wnt2b KO mice showed an accelerated rate of injury, featuring earlier immune cell infiltration and loss of differentiated epithelium compared to WT. WNT2B-deficient HIOs showed abnormal epithelial organization and an increased mesenchymal gene signature. WNT2B contributes to maintenance of the intestinal stem cell pool in mice and humans. WNT2B deficient mice, which do not have a developmental phenotype, show increased susceptibility to colonic injury but not small intestinal injury, potentially due to a higher reliance on WNT2B in the colon compared to the small intestine.WNT2B deficiency causes a developmental phenotype in human intestine with HIOs showing a decrease in their mesenchymal component and WNT2B-deficient patients showing epithelial disorganization. All RNA-Seq data will be available through online repository as indicated in Transcript profiling. Any other data will be made available upon request by emailing the study authors.</AbstractText

WNT2B Deficiency Causes Enhanced Susceptibility to Colitis Due to Increased Inflammatory Cytokine Production

Humans with WNT2B deficiency have severe intestinal disease, including significant inflammatory injury, highlighting a critical role for WNT2B. We sought to understand how WNT2B contributes to intestinal homeostasis. We investigated the intestinal health of Wnt2b knock out (KO) mice. We assessed the baseline histology and health of the small intestine and colon, as well as the impact of inflammatory challenge using dextran sodium sulfate (DSS). We also evaluated human intestinal tissue. Mice with WNT2B deficiency had normal baseline histology but enhanced susceptibility to DSS colitis due to an increased early injury response. While ISC markers were decreased, epithelial proliferation was similar to controls. Wnt2b KO mice showed an enhanced inflammatory signature after DSS treatment. Wnt2b KO colon and human WNT2B-deficient organoids both had increased levels of CXCR4 and IL6, and biopsy tissue from humans showed increased neutrophils. WNT2B is important for regulation of inflammation in the intestine. Absence of WNT2B leads to increased expression of inflammatory cytokines and increased susceptibility to gastrointestinal inflammation, particularly in the colon. [Display omitted