Mucosally transplanted mesenchymal stem cells stimulate intestinal healing by promoting angiogenesis
Nicholas A. Manieri, Madison R. Mack, Molly D. Himmelrich, Daniel L. Worthley, Elaine M. Hanson, Lars Eckmann, Timothy C. Wang, Thaddeus S. Stappenbeck
Nicholas A. Manieri, Madison R. Mack, Molly D. Himmelrich, Daniel L. Worthley, Elaine M. Hanson, Lars Eckmann, Timothy C. Wang, Thaddeus S. Stappenbeck
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Research Article Gastroenterology

Mucosally transplanted mesenchymal stem cells stimulate intestinal healing by promoting angiogenesis

Abstract

Mesenchymal stem cell (MSC) therapy is an emerging field of regenerative medicine; however, it is often unclear how these cells mediate repair. Here, we investigated the use of MSCs in the treatment of intestinal disease and modeled abnormal repair by creating focal wounds in the colonic mucosa of prostaglandin-deficient mice. These wounds developed into ulcers that infiltrated the outer intestinal wall. We determined that penetrating ulcer formation in this model resulted from increased hypoxia and smooth muscle wall necrosis. Prostaglandin I2 (PGI2) stimulated VEGF-dependent angiogenesis to prevent penetrating ulcers. Treatment of mucosally injured WT mice with a VEGFR inhibitor resulted in the development of penetrating ulcers, further demonstrating that VEGF is critical for mucosal repair. We next used this model to address the role of transplanted colonic MSCs (cMSCs) in intestinal repair. Compared with intravenously injected cMSCs, mucosally injected cMSCs more effectively prevented the development of penetrating ulcers, as they were more efficiently recruited to colonic wounds. Importantly, mucosally injected cMSCs stimulated angiogenesis in a VEGF-dependent manner. Together, our results reveal that penetrating ulcer formation results from a reduction of local angiogenesis and targeted injection of MSCs can optimize transplantation therapy. Moreover, local MSC injection has potential for treating diseases with features of abnormal angiogenesis and repair.

Authors

Nicholas A. Manieri, Madison R. Mack, Molly D. Himmelrich, Daniel L. Worthley, Elaine M. Hanson, Lars Eckmann, Timothy C. Wang, Thaddeus S. Stappenbeck

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Figure 1

PGI2 is necessary to prevent loss of α-SMA staining after mucosal injury.

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PGI2 is necessary to prevent loss of α-SMA staining after mucosal injury...
Wounds 6 days after injury in (A) WT and (B) Ptgs2–/– mice. Muscularis propria loses α-SMA staining beneath wounds in Ptgs2–/– mice within 6 days. Mechanisms leading to penetrating ulcer formation in this model are not known. m.m., muscularis mucosae (removed during biopsy); m.p., muscularis propria (containing an inner circular and outer longitudinal layer, not removed during biopsy). (C–F) Representative images of colonic sections (n = 9 wounds per group in 4 experiments) 6 days after injury from (C) WT (n = 5 mice) or (D–F) Ptgs2–/– mice treated with (C and D) vehicle (n = 6 mice), (E) 100 μg/kg PGE2 analog (n = 4 mice), or (F) 200 μg/kg PGI2 analog (n = 4 mice). Sections were stained with anti–β-catenin antisera (green, epithelium), anti–α-SMA antisera (red, smooth muscle cells), and bisbenzimide (blue, nuclei). Dashed white lines outline wound beds. Dashed yellow lines outline outer muscle (quantified in G). (G) Relative α-SMA staining in outer muscle layer underlying wounds from indicated groups of mice. n = 9 wounds per group from 4 to 6 mice per group in 4 experiments; mean ± SEM. P = 0.0039 by 1-way ANOVA. **P < 0.01, Tukey’s post-test. (H) Representative image of Ptgir–/– colon (n = 10 wounds from 6 mice in 3 experiments) 6 days after biopsy stained with anti–β-catenin antisera (green), anti–α-SMA antisera (red), and bisbenzimide (blue). (I) Relative α-SMA staining of the outer muscle layer in WT and Ptgir–/– mice 6 days after injury. Mean ± SEM. *P < 0.05, Student’s t test. WT samples: n = 6 wounds from 4 mice in 3 experiments; Ptgir–/– samples: n = 10 wounds from 6 mice in 3 experiments. Scale bar: 100 μm.