Comparative Study of The Effect of Bone Marrow-Derived Mesenchymal Stem Cells versus sulfasalazine Administration in Experimentally-induced ulcerative colitis in Rats

Abstract

SUMMARY U lcerative colitis is a disease affecting the quality of life with high morbidity and makes an economic burden. Several trials had been done to develop an appropriate treatment that cures or at least decreases the prognosis and severity of the disease. This study aimed to evaluate the possible curative role of isolated cultured BM MSCs in experimentally-induced ulcerative colitis in adult male albino rats and to compare between the histological effects of both BM MSCs and sulfasalazine in the treatment of ulcerative colitis. Five rats of average weight 100grams were used for obtaining the BM MSCs. Forty three adult male albino (Sprague-dawely strain) rats of average weight 150-180 grams were included in this study. The animals were divided into five groups: Group I: (Control group): It included 15 rats, divided into three subgroups: Subgroup Ia: Negative control. Subgroup Ib: Positive control animals that were given single dose of1ml phosphate buffered saline (PBS) via intravenous route then the rats were left for one week. Subgroup Ic: Positive control animals that were given sulfasalazine in a dose of 360 mg/kg body weight dissolved in 3ml distilled water given orally daily by intra-gastric tube for one week. Group II: (Ulcerative colitis group): It included 7 rats in which ulcerative colitis was induced by oral administration of 5ml of 2%DSS daily for 3 days using a gastric tube. This was followed by a single injection of 0.5ml of 30% ethanol at the 4th day using a neonatal Ryle tube introduced rectally up to 8 cm proximal to the anus. The animals were kept fasting 24 hours before ethanol administration. These rats were sacrificed on the 5th day from the start of the experiment to ensure the induction of ulcerative colitis. Group III: (Recovery group): It included 7 rats in which ulcerative colitis was induced as in group II. The rats were left for 12 days from the start of the experiment. Group IV: (Sulfasalazine treated group): It included 7 rats in which ulcerative colitis was induced as in group II. On the 5th day from the start of the experiment, the rats received sulfasalazine in a dose of 360 mg/kg body weight dissolved in 3ml distilled water given orally daily by intra-gastric tube for one week. Group V: (Stem cell treated group): It included 7 rats in which ulcerative colitis was induced as in group II. On the 5th day from the start of the experiment, the rats received single dose of 1ml PBS containing BM MSCs at a concentration of 1×106 cells/ml via intravenous route (tail vein) and were left for one week. In each group, the sections of the distal colon were histologically examined using Hx and E, Combined Alcian blue-PAS stain and Mallory trichrome stain. The sections were also examined immuno-histochemically using the immune peroxidase technique for detection of Tumor necrosis factor alpha (TNF α) and proliferating cell nuclear antigen (PCNA). Immuno-fluorescent technique was used to detect the labeled stem cells. Morphometric studies were done for measurement of the number of goblet cells, length of the crypts, area percentage of the alcian blue positive mucin, number of intraepithelial neutrophils and number of neutrophils in the lamina propria in the different groups. Finally statistical analysis of the findings was done using one way ANOVA to compare between the different groups. P<0.05 was considered significant.