Behavior of Cardiomyocytes and Skeletal Muscle Cells on Different Extracellular Matrix Components?Relevance for Cardiac Tissue Engineering

The purpose of this study was to investigate the tissue regenerating and biomechanical properties of processed eggshell membrane powder (PEP) for use in 3D-scaffolds. PEP is a low-cost, natural biomaterial with beneficial bioactive properties. Most importantly, this material is available as a by-product of the chicken egg processing (breaking) industry on a large scale, and it could have potential as a low-cost ingredient for therapeutic scaffolds. Scaffolds consisting of collagen alone and collagen combined with PEP were produced and analyzed for their mechanical properties and the growth of primary fibroblasts and skeletal muscle cells. Mechanical testing revealed that a PEP/collagen-based scaffold increased the mechanical hardness of the scaffold compared with a pure collagen scaffold. Scanning electron microscopy (SEM) demonstrated an interconnected porous structure for both scaffolds, and that the PEP was evenly distributed in dense clusters within the scaffold. Fibroblast and skeletal muscle cells attached, were viable and able to proliferate for 1 and 2 weeks in both scaffolds. The cell types retained their phenotypic properties expressing phenotype markers of fibroblasts (TE7, alpha-smooth muscle actin) and skeletal muscle (CD56) visualized by immunostaining. mRNA expression of the skeletal muscle markers myoD, myogenin, and fibroblasts marker (SMA) together with extracellular matrix components supported viable phenotypes and matrix-producing cells in both types of scaffolds. In conclusion, PEP is a promising low-cost, natural biomaterial for use in combination with collagen as a scaffold for 3D-tissue engineering to improve the mechanical properties and promote cellular adhesion and growth of regenerating cells.

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Recapitulating Cardiac Structure and Function In Vitro from Simple to Complex Engineering

Micromachines ◽

10.3390/mi12040386 ◽

2021 ◽

Vol 12 (4) ◽

pp. 386

Author(s):

Ana Santos ◽

Yongjun Jang ◽

Inwoo Son ◽

Jongseong Kim ◽

Yongdoo Park

Keyword(s):

Tissue Engineering ◽

Extracellular Matrix ◽

Computational Modeling ◽

Cardiac Tissue ◽

Cardiac Development ◽

Heart Tissue ◽

Cardiac Tissue Engineering ◽

Cardiac Cells

Cardiac tissue engineering aims to generate in vivo-like functional tissue for the study of cardiac development, homeostasis, and regeneration. Since the heart is composed of various types of cells and extracellular matrix with a specific microenvironment, the fabrication of cardiac tissue in vitro requires integrating technologies of cardiac cells, biomaterials, fabrication, and computational modeling to model the complexity of heart tissue. Here, we review the recent progress of engineering techniques from simple to complex for fabricating matured cardiac tissue in vitro. Advancements in cardiomyocytes, extracellular matrix, geometry, and computational modeling will be discussed based on a technology perspective and their use for preparation of functional cardiac tissue. Since the heart is a very complex system at multiscale levels, an understanding of each technique and their interactions would be highly beneficial to the development of a fully functional heart in cardiac tissue engineering.

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A Strategy to Enhance Secretion of Extracellular Matrix Components by Stem Cells: Relevance to Tissue Engineering

Tissue Engineering Part A ◽

10.1089/ten.tea.2017.0060 ◽

2018 ◽

Vol 24 (1-2) ◽

pp. 145-156 ◽

Cited By ~ 7

Author(s):

Navaneethakrishnan Krishnamoorthy ◽

Yuan‐Tsan Tseng ◽

Poornima Gajendrarao ◽

Padmini Sarathchandra ◽

Ann McCormack ◽

...

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Extracellular Matrix ◽

Extracellular Matrix Components ◽

Matrix Components

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Native cardiac environment and its impact on engineering cardiac tissue

Biomaterials Science ◽

10.1039/c8bm01348a ◽

2019 ◽

Vol 7 (9) ◽

pp. 3566-3580 ◽

Cited By ~ 9

Author(s):

Verena Schwach ◽

Robert Passier

Keyword(s):

Tissue Engineering ◽

Extracellular Matrix ◽

Embryonic Development ◽

Human Heart ◽

Cardiac Tissue ◽

Cardiac Tissue Engineering ◽

Adult Human ◽

Natural And Synthetic

In this review, we describe the progressive build-up of the cardiac extracellular matrix (ECM) during embryonic development, the ECM of the adult human heart and the application of natural and synthetic biomaterials for cardiac tissue engineering using hPSC-CMs.

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Vascularisation for cardiac tissue engineering: the extracellular matrix

Thrombosis and Haemostasis ◽

10.1160/th14-05-0480 ◽

2015 ◽

Vol 113 (03) ◽

pp. 532-547 ◽

Cited By ~ 15

Author(s):

Chinmoy Patra ◽

Aldo Boccaccini ◽

Felix Engel

Keyword(s):

Tissue Engineering ◽

Extracellular Matrix ◽

Cardiac Tissue ◽

Heart Diseases ◽

Cardiac Tissue Engineering ◽

Promising Candidate ◽

Realistic Option ◽

Extracellular Matrix Molecules ◽

Heart Muscle Cells

SummaryCardiovascular diseases present a major socio-economic burden. One major problem underlying most cardiovascular and congenital heart diseases is the irreversible loss of contractile heart muscle cells, the cardiomyocytes. To reverse damage incurred by myocardial infarction or by surgical correction of cardiac malformations, the loss of cardiac tissue with a thickness of a few millimetres needs to be compensated. A promising approach to this issue is cardiac tissue engineering. In this review we focus on the problem of in vitro vascularisation as implantation of cardiac patches consisting of more than three layers of cardiomyocytes (> 100 μm thick) already results in necrosis. We explain the need for vascularisation and elaborate on the importance to include non-myocytes in order to generate functional vascularised cardiac tissue. We discuss the potential of extracellular matrix molecules in promoting vascularisation and introduce nephronectin as an example of a new promising candidate. Finally, we discuss current biomaterial- based approaches including micropatterning, electrospinning, 3D micro-manufacturing technology and porogens. Collectively, the current literature supports the notion that cardiac tissue engineering is a realistic option for future treatment of paediatric and adult patients with cardiac disease.

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Alignment of Skeletal Muscle Cells Cultured in Collagen Gel by Mechanical and Electrical Stimulation

International Journal of Tissue Engineering ◽

10.1155/2014/621529 ◽

2014 ◽

Vol 2014 ◽

pp. 1-5 ◽

Cited By ~ 5

Author(s):

Takara Tanaka ◽

Noriko Hattori-Aramaki ◽

Ayano Sunohara ◽

Keisuke Okabe ◽

Yoshiaki Sakamoto ◽

...

Keyword(s):

Skeletal Muscle ◽

Tissue Engineering ◽

Electrical Stimulation ◽

Collagen Gel ◽

Muscle Cells ◽

Mechanical Force ◽

C2c12 Cells ◽

Skeletal Muscle Cells ◽

Collagen Gels ◽

Cells Cultured

For in vitro tissue engineering of skeletal muscle, alignment and fusion of the cultured skeletal muscle cells are required. Although the successful alignment of skeletal muscle cells cultured in collagen gel has been reported using a mechanical force, other means of aligning cultured skeletal muscle cells have not been described. However, skeletal muscle cells cultured in a two-dimensional dish have been reported to align in a uniform direction when electrically stimulated. The purpose of this study is to determine if skeletal muscle cells cultured three-dimensionally in collagen gels can be aligned by an electrical load. By adding direct current to cells of the C2C12 skeletal muscle cell line cultured in collagen gel, it was possible to align C2C12 cells in a similar direction. However, the ratio of alignment was better when mechanical force was used as the means of alignment. Thus for tissue engineering of skeletal muscle cells, electrical stimulation may be useful as a supplementary method.

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Comparison of two methods for prolong storage of decellularized mouse whole testis for tissue engineering application: An experimental study

International Journal of Reproductive BioMedicine ◽

10.18502/ijrm.v19i4.9058 ◽

2021 ◽

Author(s):

Nasrin Majidi Gharenaz ◽

Mansoureh Movahedin ◽

Zohreh Mazaheri

Keyword(s):

Tissue Engineering ◽

Experimental Study ◽

Extracellular Matrix ◽

Mouse Embryonic Fibroblast ◽

Slow Freezing ◽

Histological Evaluation ◽

Biological Scaffolds ◽

Extracellular Matrix Components ◽

Matrix Components ◽

Embryonic Fibroblast

Background: Biological scaffolds are derived by the decellularization of tissues or organs. Various biological scaffolds, such as scaffolds for the liver, lung, esophagus, dermis, and human testicles, have been produced. Their application in tissue engineering has created the need for cryopreservation processes to store these scaffolds. Objective: The aim was to compare the two methods for prolong storage testicular scaffolds. Materials and Methods: In this experimental study, 20 male NMRI mice (8 wk) were sacrificed and their testes were removed and treated with 0.5% sodium dodecyl sulfate followed by Triton X-100 0.5%. The efficiency of decellularization was determined by histology and DNA quantification. Testicular scaffolds were stored in phosphate-buffered saline solution at 4ºC or cryopreserved by programmed slow freezing followed by storage in liquid nitrogen. Masson’s trichrome staining, Alcian blue staining and immunohistochemistry, collagen assay, and glycosaminoglycan assay were done prior to and after six months of storage under each condition. Results: Hematoxylin-eosin staining showed no remnant cells after the completion of decellularization. DNA content analysis indicated that approximately 98% of the DNA was removed from the tissue (p = 0.02). Histological evaluation confirmed the preservation of extracellular matrix components in the fresh and frozen-thawed scaffolds. Extracellular matrix components were decreased by 4ºC-stored scaffolds. Cytotoxicity tests with mouse embryonic fibroblast showed that the scaffolds were biocompatible and did not have a harmful effect on the proliferation of mouse embryonic fibroblast cells. Conclusion: Our results demonstrated the superiority of the slow freezing method for prolong storage of testicular scaffolds. Key words: Cryopreservation, Testis, Scaffold, Mouse.

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