Anat Cell Biol.  2021 Sep;54(3):361-374. 10.5115/acb.21.034.

Therapeutic efficiency of adipose-derived mesenchymal stem cells in healing of experimentally induced gastric ulcers in rats

• Hassan SA
• Abou Elghait AT
• Abdelqader ZS
• Meligy FY

• KMID: 2521049
• DOI: http://doi.org/10.5115/acb.21.034

Abstract

Gastric (peptic) ulcer is a major gastrointestinal disorder with high morbidity and mortality. While several drugs have been used to treat gastric ulcers, such as proton pump inhibitor-based triple therapy for Helicobacter pylori eradication, but hey result in adverse side effects. Therefore, development of new alternative therapies is desirable. Many recent studies have shown that mesenchymal stem cells (MSCs) might have an enhancing effect on the ulcerated gastric mucosa. The aim of this study is to evaluate the efficacy of MSCs in the treatment of indomethacin-induced gastric ulcer, and to compare it with the normal ulcer autohealing. This work was performed on 36 adult male albino rats, divided into four groups: Group I (control group), Group II (ulcer group), Group III (autohealing group), and Group IV (stem cells-treated group). The histological changes of gastric mucosa were examined in sections stained with H&E using light microscope for expression of vascular endothelial growth factors (VEGF) and proliferating cell nuclear antigen (PCNA) in immunohistochemical stained sections using image analyzer. The results from MSCs-treated group revealed restoration of the normal architecture of the gastric mucosa with comparison to the autohealing group which showed excessive granulation tissue and heavy cellular infiltration with disorganized architecture of the fundic mucosa. Immunohistochemical examination showed strong expression of both VEGF and PCNA in the MSCs-treated group. So it was concluded that MSCs accelerate gastric ulcer healing when injected intraperitoneally, compared to autohealing process which showed delayed healing.

Keyword

Gastric ulcer; Mesenchymal stem cells; Indomethacin; Proliferating cell nuclear antigen; Vascular endothelial growth factors

Figure

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  Fig. 1 Macroscopic photographs of gastric mucosa from rats in the six groups showing: (A) the glandular portion (Gl) with stomach rugae and fore-stomach (F) (Group I). (B–E) Showing numerous hemorrhagic lesions in the glandular mucosa (H) (Group II); (C) Group IIIa; (D) Group IIIa; (E) Group IVa; (F) Group IVb. (G) Stastical analysis of the ulcer index. aSignificant difference from the control group (Group I) at P<0.05; bSignificant difference from the ulcer group (Group II) at P<0.05.

• 

  Fig. 2 H&E stained sections of the glandular portion showing (Group I) surface epithelium (arrowhead) and gastric pits (arrow) (A), high magnification (B, C). Group II (A) the ulcer region (double arrow) with with disfigured appearance and partial shedding of the surface epithelium (E) and upper portion of the gland and NO. (B, C) High magnification; the middle and lower part of fundic glands showing dense pyknotic nuclei (tailed arrow), cellular infilteration (INF), and nuclei of connective tissue (CT) cells (wavy arrow). Group IIIa (A, B) the glandular portion showing partial loss of the mucosa in the ulcer region (double arrow) with apparent normal mucosa (NO), cellular infilteration of the submucosa (INF) and dilated blood capillaries (BC), Disorganized fundic glands (G) with granulation tissue (thick arrow), high magnification (C). Group IIIb (A) the fundic mucosa in the ulcer region (double arrow), granulation tissue (thick arrow) with disfigured epithelial and glandular cells and heavy cellular infilteration (INF) of mucosa and submucosa (S), (B, C) high magnification shows infrequent parietal cells (P). Group IVa (A, B) fundic mucosa in the ulcer region (double arrow) with disorganized mucosa, granulation tissue (thick arrow) and cellular infiltration (INF) of submucosa. (C) High magnification. Group IVb (A) the glandular parts showing complete restoration of the epithelium (arrow head), gastric pits (arrow) and fundic glands (G), the muscularis mucosa (curved arrow) and muscularis externa (ME) appear normal. (B, C) Parietal (P) cells, mucous neck (M) cells and chief cells (C) appear normal, the parietal cells (P) and chief cells (C) appear normal. Scale bar for all photomicrographs: 25 μm.

• 

  Fig. 3 Immunostained section for proliferating cell nuclear antigen (PCNA) shows Group I (A), the expression in the epithelium (arrows), inset; Group II (B), weak PCNA antibody expression (arrows) in the ulcer region; Group IIIa (C), weak PCNA antibody expression (arrow) in the ulcer region (double arrow); Group IIIb (D), weak PCNA antibody expression (arrows) in ulcer region (double arrow); Group IVa (E), weak PCNA antibody expression (arrow) in the ulcer region (double arrow); Group IVb (F), strong PCNA expression (arrows) at the neck region. (G) Statistical analysis. aSignificant difference from the control group (Group I) at P<0.05. bSignificant difference from the ulcer group (Group II) at P<0.05. Scale bar for all photomicrographs: 25 μm.

• 

  Fig. 4 Immunostained section for vascular endothelial growth factors (VEGF) antibody expression shows Group I (A), the expression in the epithelium and the lamina propria (arrows), inset (arrow head); Group II (B), weak VEGF antibody expression (arrows) in the ulcer region (double arrow); Group IIIa (C) weak VEGF antibody expression (arrows) in the ulcer region (double arrow); Group IIIb (D), weak VEGF antibody expression (arrows) in ulcer region; Group IVa (E), weak VEGF antibody expression in the ulcer region (double arrow); Group IVb (F), strong VEGF antibody expression (arrows) in the epithelium and lamina propria of the fundic glands. Scale bar for all photomicrographs: 25 μm.

• 

  Fig. 5 Photomicrographs of cultured adipose derived mesenchymal stem cells after 24 hours of isolation showing that (A) the cells appear spindle shaped cells (arrowheads) and are accompanied by a large number of red blood cells (R). (B) Cells after 7 days of isolation appear spindle-shaped (arrowheads) with fusiform nuclei and approximately 80% to 90% confluence. Scale bar: 50 μm.

• 

  Fig. 6 The flow cytometric analysis of the isolated mesenchymal stem cells (MSCs) showing (A) control group (Isotope); (B): MSCs are positive to CD90 antibody; (C) MSCs are positive to CD271 antibody; (D) MSCs are positive to CD73 antibody; and (E) MSCs are negative to CD45 antibody.

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  Fig. 7 Images Gel electrophoresis analysis after PCR showing (A) GABDH expression on the 1st and 4th days; human leukocyte antigen (HLA) expression was down regulated in the control group on both the 1st and the 4th days. (B) GABDH expression in the stem cells-treated group on the 1st 4th days. (C) HLA expression was upregulated in the Stem cells treated group on the 1st and 4th days. (D) Statistical analysis (histogram 3). *Significant difference of the control group vs. the Stem cells treated group at P<0.05.

Reference

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