Christiano Costa Esposito

Use of butyrate or glutamine in enema solution reduces inflammation and fibrosis in experimental diversion colitis

AIM To investigate whether butyrate or glutamine enemas could diminish inflammation in experimental diversion colitis. METHODS Wistar specific pathogen-free rats were submitted to a Hartmanns end colostomy and treated with enemas containing glutamine, butyrate, or saline. Enemas were administered twice a week in the excluded segment of the colon from 4 to 12 wk after the surgical procedure. Follow-up colonoscopy was performed every 4 wk for 12 wk. The effect of treatment was evaluated using video-endoscopic and histologic scores and measuring interleukin-1β, tumor necrosis factor-alpha, and transforming growth factor beta production in organ cultures by enzyme linked immunosorbent assay. RESULTS Colonoscopies of the diverted segment showed mucosa with hyperemia, increased number of vessels, bleeding and mucus discharge. Treatment with either glutamine or butyrate induced significant reductions in both colonoscopic (P < 0.02) and histological scores (P < 0.01) and restored the densities of collagen fibers in tissue (P = 0.015; P = 0.001), the number of goblet cells (P = 0.021; P = 0.029), and the rate of apoptosis within the epithelium (P = 0.043; P = 0.011) to normal values. The high levels of cytokines in colon explants from rats with diversion colitis significantly decreased to normal values after treatment with butyrate or glutamine. CONCLUSION The improvement of experimental diversion colitis following glutamine or butyrate enemas highlights the importance of specific luminal nutrients in the homeostasis of the colonic mucosa and supports their utilization for the treatment of human diversion colitis.

Heterologous mesenchymal stem cells successfully treat femoral pseudarthrosis in rats.

BackgroundThis study evaluated the effectiveness of treating pseudarthrosis in rats by using bone marrow cell suspensions or cultures of bone marrow mesenchymal stromal cellsMethodsThirty-eight specific pathogen-free (SPF) animals were randomly assigned to four groups: Group 1, Control, without surgical intervention; Group 2 (Placebo), experimental model of femoral pseudarthrosis treated only with saline solution; Group 3, experimental model of femoral pseudarthrosis treated with heterologous bone marrow cells suspension; Group 4, experimental model of femoral pseudarthrosis treated with cultures of heterologous mesenchymal stromal cells from bone marrow. When pseudarthrosis was confirmed by simple radiological studies, digital radiography and histopathology after a 120-day postoperative period, Groups 2, 3 and 4 were treated as above. At 30, 60 and 90 days after the treatment, all animals were evaluated by simple radiological studies, and at the end of the experiment, the animals were assessed by computed axial tomography and anatomopathological and histomorphometric examinations.ResultsInjected cells were detected in the areas affected by pseudarthrosis using scintigraphy within the first 24 hours after their administration. After 60 days, the animals of Group 3 showed callus formation while the animals of Group 4 presented periosteal reaction and had some consolidated areas. In contrast, Group 2 showed a predominance of fibro-osteoid tissue. After 90 days, bone consolidation and remodeling was observed in all animals from Group 3 whereas animals from Group 4 exhibited partial consolidation and those ones from Group 2 persisted with pseudarthrosis.ConclusionThe treatment with heterologous bone marrow cells suspension proved to be effective in the treatment of pseudarthrosis whereas cultures of heterologous bone marrow mesenchymal stromal cells did not show the same potential to aid bone healing.

Mo2033 Butyrate or Glutamine Enemas Prevent Inflammation and Fibrosis in Experimental Diversion Colitis

Background and aims: Diversion colitis is a relatively common complication of colostomy involving the excluded colons segment. Although the pathogenesis of the chronic inflammatory process remains unclear, the lack of luminal nutrients is believed to play a major role in the development of the disease. Therefore, we tested whether essential colonocyte nutrients, such as butyrate or glutamine could attenuate the inflammation in a model of diversion colitis. Methods: Forty eight male Wistar SPF rats were randomly distributed in 4 groups of 12 rats each. Animals were submitted to a Hartmanns end colostomy, and treated with enemas containing glutamine, butyrate, or saline. Enemas were administered twice a week in the excluded segment of the colon from the 4th to the 12th week after the surgical procedure. Follow-up video-colonoscopy was performed every 4 weeks, during 12 weeks. Afterwards, animals of all groups, including a normal control group were sacrificed either on the 8th week or the 12th week. The effects of treatment were evaluated using videoendoscopic and histologic scores, Picrosirius, Periodic Acid of Schiff and TUNEL staining, and also measuring the production of IL-1β, TNF-α and TGF-β in organ cultures, by ELISA. Results: Colonoscopies of the diverted segment, on 8th and 12th weeks, showed mucosa with hyperemia, increased number of vessels, spontaneous bleeding and mucus discharge, with the peak of the inflammatory activity in the 8th week. Treatment with either glutamine or butyrate prevented a significant increase in both colonoscopic (P < 0.01) and histological (P < 0.01) inflammatory scores, and stabilized the densities of collagen fibers in tissue (P < 0.02), the number of goblet cells (P <0.04), and the rate of apoptosis within the epithelium (P < 0.04) to normal low values, compared to the high values of the untreated inflamed tissues. Butyrate and glutamine enemas significantly abrogated the increased production of IL-1β and TGF-β (butyrate), and also of TNF-α (butyrate and glutamine) from tissue explants of surgically diverted colon, and levels were kept within the normal ranges following treatment. Conclusion: Topical administration of either glutamine or butyrate provides mucosal protection against the development of inflammation in the diverted colonic segment after colostomy. These results strongly suggest that the lack of specific luminal nutrients can induce inflammation, and support the potential utilization of glutamine or butyrate enemas for the treatment and possibly the prevention of human diversion colitis.

Comparative study between ultrasound biomicroscopy and histopathology of diversion colitis on rats

An ultrasound biomicroscopic image system, using a 50 MHz PVDF transducer (f-number = 1.5, focal distance = 4.4 mm, -6 dB bandwidth of 31.2 MHz), was employed as a complementary method to improve the diagnosis and to characterize the course and severity of diversion colitis. Nine SPF Wistar rats, Rattus norvegicus (Berkenhout, 1769), were randomly distributed into groups I- Control, II and III-Hartmanns colostomy. Tissue samples comprised the distal colon (n=3) for group I and the excluded segment excised 8 weeks in group II (n=3) and 25 weeks in group III (n=3), after colostomy. UBM images of each sample, in rectangular fragments of 5times5 mm, were obtained. Mucosa, lamina propria, submucosa and muscular layers were identified in the UBM images and morphologically correlated with the histology of the samples from group I. UMB images from group II revealed morphologically preserved muscle layer and enlarged lamina propria and submucosa (due to an edema associated with leukocyte infiltration of the lamina propria). Extensive mucosa atrophy was found in group III. The morphological results found in ultrasound images for groups I, II and III are similar to those obtained by histological examinations.