Transplantation of Fibroblast Sheets with Blood Mononuclear Cell Culture Exerts Cardioprotective Effects by Enhancing Anti-Inflammation and Vasculogenic Potential in Rat Experimental Autoimmune Myocarditis Model.

Kaori Sekine,Masaki Miyazawa,Haruchika Masuda,Haruo Hanawa,Shinichi Matsuda,Takayuki Asahara,Tetsuro Tamaki,Akira T Kawaguchi

doi:10.3390/biology11010106

Kaori Sekine, Masaki Miyazawa + Show 6 more

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https://doi.org/10.3390/biology11010106

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Abstract

Simple SummaryFulminant myocarditis (FM) is a serious inflammatory lesion of the myocardium accompanied by cardiac dysfunction, transitioning to end-stage heart failure. Due to such a difficult pathology, a therapeutic strategy that exerts a steadfast effect has yet to be developed. Blood mononuclear cells (MNCs) have been previously shown to enhance the quality and quantity of cellular fractions (QQMNCs) with anti-inflammatory and vasculogenic potential using the one culture system. The aim of this study was to investigate whether transplantation therapy with hybrid cell sheets of fibroblasts and QQMNCs improves cardiac function in a rat model with experimental autoimmune myocarditis (EAM) induced by purified porcine cardiac myosin. The transplanted hybrid cell sheet exerts cardioprotective effects against EAM, resulting in limited left ventricular remodeling and partially improved cardiac functions due to revascularization, anti-inflammation, and anti-fibrosis. Thus, tissue engineering using hybrid cell sheets of fibroblasts constructed with QQMNCs is expected to provide an effective therapeutic option for patients with severe FM.Fulminant myocarditis causes impaired cardiac function, leading to poor prognosis and heart failure. Cell sheet engineering is an effective therapeutic option for improving cardiac function. Naïve blood mononuclear cells (MNCs) have been previously shown to enhance the quality and quantity of cellular fractions (QQMNCs) with anti-inflammatory and vasculogenic potential using the one culture system. Herein, we investigated whether autologous cell sheet transplant with QQMNCs improves cardiac function in a rat model with experimental autoimmune myocarditis (EAM). Fibroblast sheets (F-sheet), prepared from EAM rats, were co-cultured with or without QQMNCs (QQ+F sheet) on temperature-responsive dishes. QQ+F sheet induced higher expression of anti-inflammatory and vasculogenic genes (Vegf-b, Hgf, Il-10, and Mrc1/Cd206) than the F sheet. EAM rats were transplanted with either QQ+F sheet or F-sheet, and the left ventricular (LV) hemodynamic analysis was performed using cardiac catheterization. Among the three groups (QQ+F sheet, F-sheet, operation control), the QQ+F sheet transplant group showed alleviation of end-diastolic pressure–volume relationship on a volume load to the same level as that in the healthy group. Histological analysis revealed that QQ+F sheet transplantation promoted revascularization and mitigated fibrosis by limiting LV remodeling. Therefore, autologous QQMNC-modified F-sheets may be a beneficial therapeutic option for EAM.

Highlights

Fulminant myocarditis (FM) is an inflammatory disease of the myocardium that often deteriorates rapidly, culminating in end-stage heart failure (HF), which is a major cause of mortality in cardiovascular diseases [1,2]
The calculated total QQMNCs derived from 100 mL of peripheral blood decreased from original cell counts, with an average decrease of 0.31-fold in the healthy control rats and 0.28-fold in the Experimental autoimmune myocarditis (EAM) rats (Figure 2A)
To evaluate the vasculogenic potential of cultured QQMNCs compared to freshly isolated peripheral blood mononuclear cells (PBMNCs), 0.25 × 105 cells were seeded onto a methylcellulose-coated dish, and at day 7 post-seeding, pEPC-CFUs and dEPC-CFUs were separately counted (Figure 2B,C)

Summary

Introduction

Fulminant myocarditis (FM) is an inflammatory disease of the myocardium that often deteriorates rapidly, culminating in end-stage heart failure (HF), which is a major cause of mortality in cardiovascular diseases [1,2]. FM is known to be caused by various viral or bacterial infections, toxic substances, autoimmune conditions, or adverse medication effects [3]. Due to a lack of effective therapies for FM, the only option available for treatment of end-stage HF, caused by FM, is heart transplantation. There are various problems, including the shortage of heart transplantation donors, prolonged need for immunosuppression, and risk of organ rejection. Novel treatment strategies are urgently required for end-stage HF. Since the late 1990s, stem cell-based regenerative therapy has been developed as an alternative to heart transplantation [4]

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