الفهرس | Only 14 pages are availabe for public view |
Abstract This study aimed to evaluate the adverse effects of Cytarabine drug on the submandibular salivary glands, and the possible regenerative potential of systemically injected bone marrow-derived mesenchymal stem cells (BMMSCs). Materials and methods: A total of 30 male Albino Wister rats, their weight was around 200- 250gm. They were 8–12 weeks old. The rats of each group were caged in separate cages in the animal house, at Cairo University. The experimental design was that the animals were divided into three groups equally: group I: 10 rats (Control group) received intraperitoneal injection of 0.5 ml of phosphate-buffered saline (PBS) for five consecutive days. group II: 10 rats (Cytarabine group) were given a daily intraperitoneal dose injection of Cytarabine at a dose of 100 mg/kg body weight for five consecutive days. group III: 10 rats (Cytarabine and BMMSCs group) were given Cytarabine at the same previously mentioned dose. One day after the last dose of Cytarabine, the same rats received BMMSCs with a single dose of 1.5 × 106 cells per body suspended in 0.5 ml of PBS intravenously at the tail vein. After six days of administration of the BMMSCs, the animals were sacrificed under chloral hydrate anesthesia. The submandibular salivary glands were dissected out of each group. Histological procedures: The submandibular salivary glands were fixed in buffered formalin solution, then they were dehydrated, after that they were embedded in paraffin wax, and sections of about 5-6 µm were obtained. Then they were deparaffinized and stained by: 1-Hematoxylin and Eosin stain: for general histological examination. 2-Immunohistochemical staining using caspase-3 antibodies. 3-Immunofluorescence tracking for the BMMSCs that were labeled with PKH67 green fluorescent dye. Results: I-Hematoxylin and eosin stain results group I (control group): The submandibular salivary glands showed normal serous acini, normal striated ducts, and normal granular convoluted ducts. Higher magnification of the submandibular salivary glands showed normal serous acini and normal striated ducts with open-phased nuclei and basal striations. Experimental groups group II: The glands showed atrophied serous acini with an increase in the interstitial spaces. The acini showed vacuolization. The striated ducts were dilated with stagnation of the secretory material. The granular convoluted ducts lost their normal architecture and cellular arrangement. The blood vessels were found to be dilated and engorged with red blood cells. At a higher magnification, the nuclei of the striated ducts showed cytoplasmic degeneration, karyorrhexis, and pyknosis. Serous acini showed cytoplasmic vacuolization. group III: The submandibular glands showed that most of the serous acini retained their normal arrangement. The striated and granular convoluted ducts regained their normal architecture and cellular arrangement. A limited number of congested blood vessels were detected. At a higher magnification, the nuclei of the striated ductal cells restored their normal arrangement. The granular convoluted duct had a nearly normal cellular arrangement with normal nuclei position. The serous acini were normal in shape and had a normal nuclear arrangement. II-Immunohistochemical results group I (control group) The acinar and ductal cells revealed negative immunoreaction to caspase-3. Experimental groups group II The serous acini showed weak positive cytoplasmic and negative nuclear immunoreaction. the striated ductal cells showed intense positive cytoplasmic and nuclear immunoreactions. The granular convoluted ductal cells presented positive cytoplasmic and negative nuclear immunoreactions to caspase-3. group III The serous acinar cells presented negative cytoplasmic and nuclear immunoreactions. The striated and granular convoluted ducts showed moderate positive cytoplasmic and negative nuclear immunoreactions to caspase-3. III-Immunofluorescence results group I (control group) The submandibular gland’s parenchyma showed a normal structure and was free from fluorescent cells. group II The submandibular gland had lost its normal arrangement and was free from fluorescent cells. group III The submandibular gland had many fluorescent cells, this proved that the BMMSCs successfully migrated to the glands. |