Impact of decellularization on porcine myocardium as scaffold for tissue engineered heart tissue
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Decellularized myocardium has been proposed to construct tissue engineered heart tissue, providing the advantage of natural extracellular architecture. Various decellularization protocols have been developed, but the impact of individual decellularization reagent in the protocol remains unclear. The aim of this study is to evaluate the structural impact of three commonly used decellularization reagents on the porcine myocardium. We decellularized porcine heart tissue with trypsin, Triton X-100 or SDS, and analyzed the morphological characteristics of the remaining tissue by SEM, AFM and two-photon LSM. We further recellularized the scaffold with rat myocardial fibroblasts and cardiomyocytes separately. According to the H&E staining and DNA quantification, SDS decellularized more efficiently in comparison to the other two reagents. Moreover, we found distinct surface microarchitecture differences among groups. The changed structure of tissue might result in varied proliferation myocardial fibroblasts and biophysical performance of the engineered heart tissue. This study demonstrated that the microstructure of decellularized porcine heart tissue vary with decellularization agents. Compared to trypsin and Triton X-100, SDS not only decellularized more efficiently but also preserved the biocompatible microstructure of ECM for recellularization.
KeywordsSodium Dodecyl Sulfate Heart Tissue Phosphate Buffer Saline Solution Engineer Heart Tissue Decellularization Process
We are indebted to W. Bian from the Shanghai Institute of Biochemistry and Cell Biology (SIBCB) for help with two-photon LSM and to S. L. Wang from East China University of Science and Technology for help with AFM.
Compliance with ethical standards
The research was partially financially supported by the National Natural Science Foundation of China (Grant Nos. 81000096, 81200093, 31330029 and 81571826).
Supplementary material 1 (MOV 4307 kb) Representative video of the rCMs seeded heart tissue, which is decellularized with Trypsin
Supplementary material 2 (MOV 4685 kb) Representative video of the rCMs seeded heart tissue, which is decellularized with SDS
Supplementary material 3 (MOV 4908 kb) Representative video of the rCMs seeded heart tissue, which is decellularized with Triton X-100
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