Innovative Biotechnology for Generation of Cardiac Tissue

Abstract

Heart transplantation remains the only curative treatment for end-stage heart failure. This life-saving option continues to be limited by the low number of organ donors, graft rejection and adverse effects of immunosuppressants. Engineering bioartificial hearts from acellular native-derived scaffolds and stem cells has gained attention because of its potential to overcome these limitations. In this study, rat hearts (n = 20) were decellularized by means of coronary perfusion with 1% sodium dodecyl sulfate (SDS) in a modified Langendorff device. The electrical field behavior of the SDS molecule was studied and it was assumed that when applying an alternating current, the exposure time of the tissue to the detergent might decrease. To repopulate the decellularized extracellular matrix (ECM), human mesenchymal stem cells (hMSCs) were used, induced to differentiate into cardiomyocytes (CMs) with 5-azacytidine (5-aza). The results showed no cellular debris and an intact ECM following decellularization. Decellularization in the presence of an electric field proved to be faster, decreasing the potential risk of ECM damage due to the detergent. After cell seeding and culturing of eight scaffolds with hMSCs, the recellularization process was analyzed using optic microscopy (OM), which showed cells suggestive for CMs. This study presents a novel and efficient decellularization protocol using an electric field and suggests that hMSCs can be useful in the generation of a bioartificial heart.

Decellularized Extracellular Matrix Scaffolds for Cardiovascular Tissue Engineering: Current Techniques and Challenges

“…This process generally involves passing a decellularization agent via the intrinsic heart vasculature, washing out the cellular components [ 11 , 127 , 128 , 129 , 130 ]. In previous work, we obtained an optimal decellularization protocol for rat hearts using a modified Langendorff experimental device (constant perfusion pressure of 80 mmHg) in the presence of an alternating rectangular electric field (20 kHz frequency and 100 mA amplitude, corresponding to 7.14 mA/cm 2 current density) [ 131 ]. Akhyari et al compared three previous protocols with a newly developed fourth protocol which implied the decellularization of whole murine hearts through coronary perfusion.…”

Whole-Heart Tissue Engineering and Cardiac Patches: Challenges and Promises

“…Numerous studies have been worked on decreasing detergent concentration and detergent exposure time to ensure that the ECM is well-preserved. In two studies, the detergent exposure time was decreased by using an electric field in the decellularization protocol [89] and using a pulsatile perfusion system since the scaffolds developed by the pulsatile perfusion showed significantly lower residual DNA content in comparison with the other scaffold developed with non-pulsatile perfusion [90]. In another study, Dal Sasso et al used protease inhibition, antioxidation, and excitation to reduce the concentration and incubation time with cytotoxic detergents as well [91].…”

Optimization of Complete Rat Heart Decellularization Using Artificial Neural Networks

“…Heart decellularization creates acellular myocardial scaffolds that perfectly mimic the native architecture, challenging to achieve with synthetic materials. Rat hearts were decellularized as described in our previous study [17]. The protocol provided an alternative electric field system, apparatus, and method of use to obtain decellularized hearts.…”

“…Rat heart decellularization was performed using an alternating electric field protocol and 1% sodium dodecyl sulfate (SDS) with good results, already described in our previous work. The optimized decellularization method ensures undamaged extracellular matrix (ECM) by applying a low detergent concentration for a short time [17].…”

“…In brief, the study analyzed the release of DNA and proteins from the heart during decellularization. The assumption was that when the concentration of the analytes became constant, the process was complete as no additional cellular material was being released [17,18]. This method of validating decellularization is harmless for the organ and allows later recellularization.…”