Hepatol Res. 2014 Oct;44(10):E206-17.

Canine mesenchymal stem cells show antioxidant properties against thioacetamide-induced liver injury in vitro and in vivo.

Quintanilha LF, Takami T, Hirose Y, Fujisawa K, Murata Y, Yamamoto N, Goldenberg RC, Terai S, Sakaida I.

Department of Gastroenterology and Hepatology, Yamaguchi University Graduate School of Medicine, Ube, Japan; Carlos Chagas Filho Biophysics Institute, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.

 

Abstract

AIM: To overcome current limitations of therapy for liver diseases, cell-based therapies using mesenchymal stem cells (MSC) have been attempted through basic and clinical approaches. Oxidative stress is a crucial factor in hepatology, and reactive oxygen species (ROS) are well-established molecules responsible for its deleterious effects. The antioxidant properties of MSC were recently demonstrated, and therefore we examined the antioxidant activity of canine MSC (cMSC), their effects on isolated hepatocytes in vitro and their curative potential against thioacetamide (TAA)-induced liver injury in vivo.

METHODS: To evaluate the ability of cMSC to challenge oxidative stress, cell viability, cytotoxicity and ROS were measured in cultured cMSC treated with TAA. Also, cMSC were co-cultured with hepatocytes in the same injury condition, and the ROS level was measured exclusively in hepatocytes. Finally, to verify the curative potential of cMSC, 2.0 × 10(6) cells or phosphate-buffered saline were injected systemically in non-obese diabetic/severe combined immunodeficiency mice that received TAA injections twice a week for 13 weeks. We then evaluated histological parameters, serum injury markers and redox homeostasis.

RESULTS: cMSC overcame TAA-induced oxidative stress in vitro, as shown by increased viability and lower cytotoxicity and ROS levels. Moreover, hepatocytes co-cultured with cMSC also showed decreased cellular ROS. The in vivo study showed that mice treated with cMSC presented with an ameliorated histological pattern, suppressed fibrosis, lower serum injury marker levels and better oxidative parameters.

CONCLUSION: We concluded that cMSC injection reduce TAA-induced liver injury through antioxidant activities and hepatoprotective effects, showing a curative potential in liver diseases.

KEYWORDS: NF-E2-related factor 2; liver; mesenchymal stem cells; oxidative stress; reactive oxygen species

PMID: 23889977

 

Supplements:

Our team has been focusing on bone marrow cells (BMCs) as a cell source for use in liver regeneration therapy and, therefore, basic and clinical research studies have been conducted (1). In our previous animal studies, we have reported that non-cultured whole BMCs infused via a peripheral vein efficiently repopulate the cirrhotic liver. Repopulated BMCs produce collagenases including matrix metalloproteinase-9. As a result, we observed reduced liver fibrosis, elevated serum albumin levels, and a significant increase in survival (2). Based on these data, we have begun “Autologous bone marrow cell infusion (ABMi) therapy” using non-cultured autologous whole BMCs (3). This therapy was officially approved as “Advanced medical technology B” in Japan. However, ABMi therapy involves BM aspiration under general anesthesia. We therefore developed a less invasive liver regeneration therapy using cultured autologous mesenchymal stem cells (MSCs) isolated from a small amount of BM fluid aspirated under local anesthesia. We reported that peripheral infusion of cultured bone marrow derived MSCs (BMSCs) reduces hepatic fibrosis in the immunodeficient cirrhotic murine model (4), consistent with the maintenance of redox homeostasis in this featured paper. In other words, we recently showed safety, efficiency and proposed a mechanism of how cultured BMSCs could improve liver diseases, which is a very important concept for basing the development of further studies. Moreover, after we confirmed the safety of this approach using middle-large animals (for example, blood examinations in Table 1, evaluations of histology and computed tomography in Figure 1), we have just started this therapy using cultured autologous BMSCs for decompensated liver cirrhotic patients after the regulatory approval in Japan (ClinicalTrials.gov; No. NCT02327832).

 

References

(1) Takami T, et al. Terai S, Sakaida I. Stem cell therapy in chronic liver disease. Curr Opin Gastroenterol 2012;28:203-208.

(2) Sakaida I, Terai S, Yamamoto N, Aoyama K, Ishikawa T, Nishina H, Okita K.

Transplantation of bone marrow cells reduces CCl4-induced liver fibrosis in mice. Hepatology 2004;40:1304-1311.

(3) Terai S, Ishikawa T, Omori K, Aoyama K, Marumoto Y, Urata Y, Yokoyama Y, et al. Improved liver function in patients with liver cirrhosis after autologous bone marrow cell infusion therapy. Stem Cells 2006;24:2292-2298.

(4) Tanimoto H, Terai S, Taro T, Murata Y, Fujisawa K, Yamamoto N, Sakaida I. Improvement of liver fibrosis by infusion of cultured cells derived from human bone marrow. Cell Tissue Res 2013;354:717-728.

 

 Table 1. The safety study about the infusion of cultured autologous canine bone marrow derived mesenchymal stem cells via a peripheral vein in healthy subjectsTT tab1

 

 

 

TT fig1

Figure 1. The safety evaluations using histological staining and Computed tomography (CT) in canines after the infusion of cultured autologous bone marrow derived mesenchymal stem cells via a peripheral vein. (A) H&E staining showed normal tissue architecture with no evidence of infarction and inflammation. (B) Sirius red staining showed normal distribution of liver fibrosis. (C) CT examinations showed any evidence of embolism in lung arteries.

 

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