Epicardial application of cardiac progenitor cells in a 3D-printed gelatin/hyaluronic acid patch preserves cardiac function after myocardial infarction
Introduction
An ischemic event in the myocardium has dire consequences for the heart since the death of cardiomyocytes within the infarcted area leaves the heart with less contractile elements. This shifts the pumping burden to the remaining viable myocardium, which can have deleterious consequences for patients suffering from the disease, potentially leading to the development of heart failure. Current therapeutic approaches do not provide patients with ways to repair the organ and instead focus on limiting secondary damages to slow down the progression of the disease [1], [2]. Therefore, end-stage heart failure patients often require heart transplantation, for which donor hearts are in short supply and carry risks of rejection [3]. Although the existence of endogenous stem cell populations in the heart has been documented [4], [5], these cells are unable to sufficiently repair the injury and restore the function of the heart.
The need to replenish the heart with new myocytes is of critical importance to avoid the chronic manifestation of the disease. In this regard, stem cell transplantation therapy offers a new therapeutic avenue to create de novo myocardium either in vitro or in vivo. A wide variety of stem cells have shown the ability to differentiate into cardiomyocytes. Of these, the multipotent Sca1+ cardiac-derived cardiomyocyte progenitor cells (CMPCs) offers a desirable combination of a patient-specific cell source with cardiogenic potential, both in vitro as in vivo [6], [7]. Besides direct involvement in tissue repair, the plethora of factors secreted by CMPCs can also activate endogenous stem cell pools, thereby making it a well suited cell type for the implementation of cardiac regenerative strategies [8], [9].
Although promising results have come out of pre-clinical and clinical studies [10], [11], [12], cardiac stem cell therapy still suffers from inefficient delivery, engraftment, and differentiation of cells in the myocardium [13], [14], [15]. Furthermore, during the progression of heart failure, extracellular matrix is also modified and replaced by scar tissue. Therefore, combining procedures aiming at regenerating both myocardial cells and the extracellular matrix could improve the effectiveness of cellular therapy. For this reason, hybrid therapies that include biomaterials and cells are being developed as potentially new therapeutic approaches for repairing myocardial tissue [15], [16]. We recently showed that tissue printing technology can be used with alginate in combination with CMPCs to create, in vitro, a cardiogenic patch with precise pore size and microstructure, which allows for better cell viability over time [17].
Here, we improved this tissue engineering approach by creating a cardiogenic scaffold of hCMPCs and a hyaluronic acid/gelatin (HA/gel) based biomaterial. The new biomaterial enhanced cell attachment and survival, and is suitable for tissue printing technology applications. We were able to build a customized patch that can harbor CMPCs without affecting their growth and differentiation potential. Furthermore, we demonstrated the in vivo applicability of the printed patch in a murine model of MI. The transplanted biocomplex sustained cell viability, leading to excellent cell survival and engraftment. Lastly, mice which received the scaffold showed improved cardiac function after MI.
Section snippets
CMPC isolation and culture
Human fetal CMPCs were isolated by magnetic cell sorting based on Sca-1 positive selection and propagated as previously described [7]. Briefly, cells were plated at 0.1% gelatin coated wells in growth medium consisting of 25% EGM-2 (3% EGM-2 single quotes (Cambrex) in EBM-2 (Cambrex)) and 75% M199 (BioWhittaker), 10% FBS (Hyclone), 1× MEM non-essential amino acids (BioWhittaker) and 1× penicillin/streptomycin. Differentiation medium consisted of 50% IMDM (GIBCO), 50% HamF12 GlutaMAX-1 (GIBCO),
3D tissue printing
Using a 3D Tissue printer Bioscaffolder, we were able to generate a biocomplex consisting of CMPCs in HA/gel matrix. The construct had a final size of 2 cm × 2 cm and 400 μm tick (Fig. 1A) CMPCs were homogenously present throughout the biocomplex (Fig. 1B–C–F). Live-dead assay was performed 2 h after printing and showed that the biocomplex generation and printing process did not affect cell viability (Fig. 1D–E).
Biocomplex in vitro characterization
Expression of cardiac markers were analyzed by PCR analysis at 1 day, and 1 and 4
Discussion
Recent evidence has accumulated showing that stem cells delivered to the heart have limited retention and poor survival in the myocardium [13], [14], [22]. The cells can fail to engraft and can be washed out by lymphatic or vascular channels. For those that are retained, the new environmental surroundings are often detrimental to their viability and may not be inductive for the most optimal therapeutic cellular response. In this respect, tissue engineering has proven to be an effective method
Conclusion
In this study, we developed an effective and translation approach to enhance hCMPC delivery to the heart. We showed that the HA/Gel matrix can be used with tissue printing technology, it allows hCMPC attachment and proliferation, and it has potential for clinic translation. After printing, hCMPCs retain their cardiogenic phenotype in vitro up to 1 month and are able to differentiate in the appropriate condition. The attachment of the cardiogenic 3D-printed patch preserved heart function by
Acknowledgments
This research forms part of the Project P1.04 SMARTCARE of the BioMedical Materials institute, co-funded by the Dutch Ministry of Economic Affairs, Agriculture and Innovation, and the Netherlands CardioVascular Research Initiative (CVON): the Dutch Heart Foundation, Dutch Federation of University Medical Centers, the Netherlands Organization for Health Research and Development, and the Royal Netherlands Academy of Sciences. The financial contribution of the Dutch Heart Foundation, the Cenci
References (40)
- et al.
Cardiac stem cell therapy and the promise of heart regeneration
Cell Stem Cell
(2013) - et al.
Cardiac stem cells in patients with ischaemic cardiomyopathy (SCIPIO): initial results of a randomised phase 1 trial
Lancet
(2011) - et al.
Intracoronary cardiosphere-derived cells for heart regeneration after myocardial infarction (CADUCEUS): a prospective, randomised phase 1 trial
Lancet
(2012) - et al.
Noninvasive quantification and optimization of acute cell retention by in vivo positron emission tomography after intramyocardial cardiac-derived stem cell delivery
J. Am. Coll. Cardiol.
(2009) - et al.
Cardiac tissue engineering using tissue printing technology and human cardiac progenitor cells
Biomaterials
(2012) - et al.
Catheter-deliverable hydrogel derived from decellularized ventricular extracellular matrix increases endogenous cardiomyocytes and preserves cardiac function post-myocardial infarction
J. Am. Coll. Cardiol.
(2012) - et al.
Materials science and tissue engineering: repairing the heart
Mayo Clin. Proc.
(2013) - et al.
Cardiac repair achieved by bone marrow mesenchymal stem cells/silk fibroin/hyaluronic acid patches in a rat of myocardial infarction model
Biomaterials
(2012) - et al.
The effect of matrix stiffness of injectable hydrogels on the preservation of cardiac function after a heart attack
Biomaterials
(2014) - et al.
Matrix elasticity directs stem cell lineage specification
Cell
(2006)
Electrical stimulation of human embryonic stem cells: cardiac differentiation and the generation of reactive oxygen species
Exp. Cell. Res.
Bioluminescence imaging of cardiomyogenic and vascular differentiation of cardiac and subcutaneous adipose tissue-derived progenitor cells in fibrin patches in a myocardium infarct model
Int. J. Cardiol.
Heart regeneration
Nature
Dissecting the molecular relationship among various cardiogenic progenitor cells
Circ. Res.
Human cardiac stem cells
Proc. Natl. Acad. Sci. U. S. A.
TGF-beta1 induces efficient differentiation of human cardiomyocyte progenitor cells into functional cardiomyocytes in vitro
Stem Cell Res.
Human cardiomyocyte progenitor cells differentiate into functional mature cardiomyocytes: an in vitro model for studying human cardiac physiology and pathophysiology
Nat. Protoc.
Relative roles of direct regeneration versus paracrine effects of human cardiosphere-derived cells transplanted into infarcted mice
Circ. Res.
Cardiomyocyte progenitor cell-derived exosomes stimulate migration of endothelial cells
J. Cell Mol. Med.
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These authors contributed equally to this work.