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Allogeneic guinea pig mesenchymal stem cells ameliorate neurological changes in experimental colitis.

Stavely R, Robinson AM, Miller S, Boyd R, Sakkal S, Nurgali K - Stem Cell Res Ther (2015)

Bottom Line: This study aims to isolate and characterise guinea pig MSCs and then test their therapeutic potential for the treatment of enteric neuropathy associated with intestinal inflammation.MSCs from both sources secreted TGF-β1 which exerted neuroprotective effects in vitro.In vitro characteristics of MSCs cannot be extrapolated to their therapeutic efficacy.

View Article: PubMed Central - PubMed

Affiliation: Centre for Chronic Disease, College of Health and Biomedicine, Western Centre for Health, Research and Education, Sunshine Hospital, 176 Furlong road, Melbourne, 3021, Victoria, Australia. rhian.stavely@live.vu.edu.au.

ABSTRACT

Background: The use of mesenchymal stem cells (MSCs) to treat inflammatory bowel disease (IBD) is of great interest because of their immunomodulatory properties. Damage to the enteric nervous system (ENS) is implicated in IBD pathophysiology and disease progression. The most commonly used model to study inflammation-induced changes to the ENS is 2,4,6-trinitrobenzene-sulfonate acid (TNBS)-induced colitis in guinea pigs; however, no studies using guinea pig MSCs in colitis have been performed. This study aims to isolate and characterise guinea pig MSCs and then test their therapeutic potential for the treatment of enteric neuropathy associated with intestinal inflammation.

Methods: MSCs from guinea pig bone marrow and adipose tissue were isolated and characterised in vitro. In in vivo experiments, guinea pigs received either TNBS for the induction of colitis or sham treatment by enema. MSCs were administered at a dose of 1 × 10(6) cells via enema 3 h after the induction of colitis. Colon tissues were collected 24 and 72 h after TNBS administration to assess the level of inflammation and damage to the ENS. The secretion of transforming growth factor-β1 (TGF-β1) was analysed in MSC conditioned medium by flow cytometry.

Results: Cells isolated from both sources were adherent to plastic, multipotent and expressed some human MSC surface markers. In vitro characterisation revealed distinct differences in growth kinetics, clonogenicity and cell morphology between MSC types. In an in vivo model of TNBS-induced colitis, guinea pig bone marrow MSCs were comparatively more efficacious than adipose tissue MSCs in attenuating weight loss, colonic tissue damage and leukocyte infiltration into the mucosa and myenteric plexus. MSCs from both sources were equally neuroprotective in the amelioration of enteric neuronal loss and changes to the neurochemical coding of neuronal subpopulations. MSCs from both sources secreted TGF-β1 which exerted neuroprotective effects in vitro.

Conclusions: This study is the first evaluating the functional capacity of guinea pig bone marrow and adipose tissue-derived MSCs and providing evidence of their neuroprotective value in an animal model of colitis. In vitro characteristics of MSCs cannot be extrapolated to their therapeutic efficacy. TGF-β1 released by both types of MSCs might have contributed to the attenuation of enteric neuropathy associated with colitis.

No MeSH data available.


Related in: MedlinePlus

Effects of guinea pig MSCs on nitrergic myenteric neurons. a–d′ Nitrergic (nNOS-IR) neurons were visualised in the myenteric plexus at 24 h (a–d) and 72 h (a′–d′). Scale bar = 50 μm. The total number of nNOS-IR neurons (e) and the proportion of nNOS-IR neurons to the total number of HuC/D-IR neurons (f) were quantified within a 2-mm2 area of the myenteric plexus in the guinea pig colon. *P <0.05, **P <0.01, ***P <0.001, n = 4 animals per group per time point. gpAT-MSC guinea pig adipose tissue-derived mesenchymal stem cell, gpBM-MSC guinea pig bone marrow-derived mesenchymal stem cell, MSC mesenchymal stem cell, TNBS 2,4,6-Trinitrobenzene sulfonic acid
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Fig8: Effects of guinea pig MSCs on nitrergic myenteric neurons. a–d′ Nitrergic (nNOS-IR) neurons were visualised in the myenteric plexus at 24 h (a–d) and 72 h (a′–d′). Scale bar = 50 μm. The total number of nNOS-IR neurons (e) and the proportion of nNOS-IR neurons to the total number of HuC/D-IR neurons (f) were quantified within a 2-mm2 area of the myenteric plexus in the guinea pig colon. *P <0.05, **P <0.01, ***P <0.001, n = 4 animals per group per time point. gpAT-MSC guinea pig adipose tissue-derived mesenchymal stem cell, gpBM-MSC guinea pig bone marrow-derived mesenchymal stem cell, MSC mesenchymal stem cell, TNBS 2,4,6-Trinitrobenzene sulfonic acid

Mentions: The two major subpopulations of neurons in the myenteric plexus, inhibitory and excitatory muscle motor and interneurons, were investigated [74]. Inhibitory neurons were labelled with anti-nNOS antibody (Fig. 8a–d′), and the total number of nNOS-IR neurons was quantified per 2-mm2 area (Fig. 8e and Table 3, n = 4 animals per group per time point). The number of nNOS-IR neurons was increased in the myenteric plexus from TNBS groups compared with sham at 24 h (P <0.01) and 72 h (P <0.05). The proportion of nNOS-IR neurons to the total number of Hu-IR neurons was increased in TNBS groups at both time points compared with sham (24 and 72 h, P <0.001) (Fig. 8f, n = 4 animals per group per time point). Both MSC treatments ameliorated the increase in the total number (gpBM-MSC: 24 and 72 h, P <0.05 and gpAT-MSC: 24 and 72 h, P <0.01) and proportion of nNOS-IR neurons at all time points (gpBM-MSC: 24 and 72 h, P <0.01; gpAT-MSCs: 24 h, P <0.001 and 72 h, P <0.01) (Fig. 8e, f and Table 3).Fig. 8


Allogeneic guinea pig mesenchymal stem cells ameliorate neurological changes in experimental colitis.

Stavely R, Robinson AM, Miller S, Boyd R, Sakkal S, Nurgali K - Stem Cell Res Ther (2015)

Effects of guinea pig MSCs on nitrergic myenteric neurons. a–d′ Nitrergic (nNOS-IR) neurons were visualised in the myenteric plexus at 24 h (a–d) and 72 h (a′–d′). Scale bar = 50 μm. The total number of nNOS-IR neurons (e) and the proportion of nNOS-IR neurons to the total number of HuC/D-IR neurons (f) were quantified within a 2-mm2 area of the myenteric plexus in the guinea pig colon. *P <0.05, **P <0.01, ***P <0.001, n = 4 animals per group per time point. gpAT-MSC guinea pig adipose tissue-derived mesenchymal stem cell, gpBM-MSC guinea pig bone marrow-derived mesenchymal stem cell, MSC mesenchymal stem cell, TNBS 2,4,6-Trinitrobenzene sulfonic acid
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4697327&req=5

Fig8: Effects of guinea pig MSCs on nitrergic myenteric neurons. a–d′ Nitrergic (nNOS-IR) neurons were visualised in the myenteric plexus at 24 h (a–d) and 72 h (a′–d′). Scale bar = 50 μm. The total number of nNOS-IR neurons (e) and the proportion of nNOS-IR neurons to the total number of HuC/D-IR neurons (f) were quantified within a 2-mm2 area of the myenteric plexus in the guinea pig colon. *P <0.05, **P <0.01, ***P <0.001, n = 4 animals per group per time point. gpAT-MSC guinea pig adipose tissue-derived mesenchymal stem cell, gpBM-MSC guinea pig bone marrow-derived mesenchymal stem cell, MSC mesenchymal stem cell, TNBS 2,4,6-Trinitrobenzene sulfonic acid
Mentions: The two major subpopulations of neurons in the myenteric plexus, inhibitory and excitatory muscle motor and interneurons, were investigated [74]. Inhibitory neurons were labelled with anti-nNOS antibody (Fig. 8a–d′), and the total number of nNOS-IR neurons was quantified per 2-mm2 area (Fig. 8e and Table 3, n = 4 animals per group per time point). The number of nNOS-IR neurons was increased in the myenteric plexus from TNBS groups compared with sham at 24 h (P <0.01) and 72 h (P <0.05). The proportion of nNOS-IR neurons to the total number of Hu-IR neurons was increased in TNBS groups at both time points compared with sham (24 and 72 h, P <0.001) (Fig. 8f, n = 4 animals per group per time point). Both MSC treatments ameliorated the increase in the total number (gpBM-MSC: 24 and 72 h, P <0.05 and gpAT-MSC: 24 and 72 h, P <0.01) and proportion of nNOS-IR neurons at all time points (gpBM-MSC: 24 and 72 h, P <0.01; gpAT-MSCs: 24 h, P <0.001 and 72 h, P <0.01) (Fig. 8e, f and Table 3).Fig. 8

Bottom Line: This study aims to isolate and characterise guinea pig MSCs and then test their therapeutic potential for the treatment of enteric neuropathy associated with intestinal inflammation.MSCs from both sources secreted TGF-β1 which exerted neuroprotective effects in vitro.In vitro characteristics of MSCs cannot be extrapolated to their therapeutic efficacy.

View Article: PubMed Central - PubMed

Affiliation: Centre for Chronic Disease, College of Health and Biomedicine, Western Centre for Health, Research and Education, Sunshine Hospital, 176 Furlong road, Melbourne, 3021, Victoria, Australia. rhian.stavely@live.vu.edu.au.

ABSTRACT

Background: The use of mesenchymal stem cells (MSCs) to treat inflammatory bowel disease (IBD) is of great interest because of their immunomodulatory properties. Damage to the enteric nervous system (ENS) is implicated in IBD pathophysiology and disease progression. The most commonly used model to study inflammation-induced changes to the ENS is 2,4,6-trinitrobenzene-sulfonate acid (TNBS)-induced colitis in guinea pigs; however, no studies using guinea pig MSCs in colitis have been performed. This study aims to isolate and characterise guinea pig MSCs and then test their therapeutic potential for the treatment of enteric neuropathy associated with intestinal inflammation.

Methods: MSCs from guinea pig bone marrow and adipose tissue were isolated and characterised in vitro. In in vivo experiments, guinea pigs received either TNBS for the induction of colitis or sham treatment by enema. MSCs were administered at a dose of 1 × 10(6) cells via enema 3 h after the induction of colitis. Colon tissues were collected 24 and 72 h after TNBS administration to assess the level of inflammation and damage to the ENS. The secretion of transforming growth factor-β1 (TGF-β1) was analysed in MSC conditioned medium by flow cytometry.

Results: Cells isolated from both sources were adherent to plastic, multipotent and expressed some human MSC surface markers. In vitro characterisation revealed distinct differences in growth kinetics, clonogenicity and cell morphology between MSC types. In an in vivo model of TNBS-induced colitis, guinea pig bone marrow MSCs were comparatively more efficacious than adipose tissue MSCs in attenuating weight loss, colonic tissue damage and leukocyte infiltration into the mucosa and myenteric plexus. MSCs from both sources were equally neuroprotective in the amelioration of enteric neuronal loss and changes to the neurochemical coding of neuronal subpopulations. MSCs from both sources secreted TGF-β1 which exerted neuroprotective effects in vitro.

Conclusions: This study is the first evaluating the functional capacity of guinea pig bone marrow and adipose tissue-derived MSCs and providing evidence of their neuroprotective value in an animal model of colitis. In vitro characteristics of MSCs cannot be extrapolated to their therapeutic efficacy. TGF-β1 released by both types of MSCs might have contributed to the attenuation of enteric neuropathy associated with colitis.

No MeSH data available.


Related in: MedlinePlus