Development of a new decellularization protocol for the whole porcine heart

Delgado, Ana Lídia Jacintho; Carreira, Ana Cláudia Oliveira; Carvalho, Hianka Jasmyne Costa de; Palma, Renata Kelly da; Sasahara, Taís Harumi de Castro; Carvalho, Carlos Augusto Brandão de; León, Marisol; Barreto, Rodrigo Egydio; Miglino, María Angélica

doi:10.18053/jctres.07.202104.017

Cited by 5 publications

(3 citation statements)

References 20 publications

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“…MTC staining was utilized for the purpose of identifying connective tissues, mainly collagen fibers (figure 4). The mechanical characteristics and elasticity of the scaffolds can be impacted by collagen proteins in the ECM of decellularized tissues, as these proteins also serve as signaling molecules for cell differentiation and proliferation [33]. Dark blue staining revealed collagen fibers, which were densely observed in the native thyroid tissue along with cell nuclei (figure 4(a)).…”

Section: Histological Analysismentioning

confidence: 99%

Optimizing decellularization protocols for human thyroid tissues: a step towards tissue engineering and transplantation

Karabıyık Acar,

Bozdağ,

Hacıhasanoğlu

et al. 2024

Biomed. Mater.

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Hypothyroidism is caused by insufficient stimulation or disruption of the thyroid. However, the drawbacks of thyroid transplantation have led to the search for new treatments. Decellularization allows tissue transplants to maintain their biomimetic structures while preserving cell adhesion, proliferation, and differentiation. This study aimed to decellularize human thyroid tissues using a structure-preserving optimization strategy and present preliminary data on recellularization. Nine methods were used for physical and chemical decellularization. Quantitative and immunohistochemical analyses were performed to investigate the DNA and extracellular matrix components of the tissues. Biomechanical properties were determined by compression test, and cell viability was examined after seeding MDA-T32 papillary thyroid cancer (PTC) cells onto the decellularized tissues. Decellularized tissues exhibited a notable decrease (<50 ng mg−1 DNA, except for Groups 2 and 7) compared to the native thyroid tissue. Nonetheless, collagen and glycosaminoglycans were shown to be conserved in all decellularized tissues. Laminin and fibronectin were preserved at comparatively higher levels, and Young’s modulus was elevated when decellularization included SDS. It was observed that the strain value in Group 1 (1.63 ± 0.14 MPa) was significantly greater than that in the decellularized tissues between Groups 2–9, ranging from 0.13 ± 0.03–0.72 ± 0.29 MPa. Finally, viability assessment demonstrated that PTC cells within the recellularized tissue groups successfully attached to the 3D scaffolds and sustained metabolic activity throughout the incubation period. We successfully established a decellularization optimization for human thyroid tissues, which has potential applications in tissue engineering and transplantation research. Our next goal is to conduct recellularization using the methods utilized in Group 1 and transplant the primary thyroid follicular cell-seeded tissues into an in vivo animal model, particularly due to their remarkable 3D structural preservation and cell adhesion-promoting properties.

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Section: Histological Analysismentioning

confidence: 99%

Optimizing decellularization protocols for human thyroid tissues: a step towards tissue engineering and transplantation

Karabıyık Acar,

Bozdağ,

Hacıhasanoğlu

et al. 2024

Biomed. Mater.

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“…Immunohistochemistry staining for elastin, fibronectin, laminin, hyaluronic acid, and heparan sulfate was performed by Methe et al in 2014 to prove that the decellularization process did not distort the structure of ECM components [ 115 ]. Positive immunofluorescence for collagen type I and elastin indicated maintenance of the mechanical properties and elasticity of the acellular cardiac scaffold [ 139 ]. The acellular state of decellularized heart biopsies could be confirmed using DAPI (4′,6-diamidino-2-phenylindole, dihydrochloride) staining with no evidence of cell nuclei or residue of nuclear material [ 11 , 115 , 139 , 140 ].…”

Section: Quantification Of Complete Decellularizationmentioning

confidence: 99%

“…Positive immunofluorescence for collagen type I and elastin indicated maintenance of the mechanical properties and elasticity of the acellular cardiac scaffold [ 139 ]. The acellular state of decellularized heart biopsies could be confirmed using DAPI (4′,6-diamidino-2-phenylindole, dihydrochloride) staining with no evidence of cell nuclei or residue of nuclear material [ 11 , 115 , 139 , 140 ]. DNA quantification is an effective technique to describe the removal of genetic material.…”

Section: Quantification Of Complete Decellularizationmentioning

confidence: 99%

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

Barbulescu

Bojin

Ordodi

et al. 2022

IJMS

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Cardiovascular diseases are the leading cause of global mortality. Over the past two decades, researchers have tried to provide novel solutions for end-stage heart failure to address cardiac transplantation hurdles such as donor organ shortage, chronic rejection, and life-long immunosuppression. Cardiac decellularized extracellular matrix (dECM) has been widely explored as a promising approach in tissue-regenerative medicine because of its remarkable similarity to the original tissue. Optimized decellularization protocols combining physical, chemical, and enzymatic agents have been developed to obtain the perfect balance between cell removal, ECM composition, and function maintenance. However, proper assessment of decellularized tissue composition is still needed before clinical translation. Recellularizing the acellular scaffold with organ-specific cells and evaluating the extent of cardiomyocyte repopulation is also challenging. This review aims to discuss the existing literature on decellularized cardiac scaffolds, especially on the advantages and methods of preparation, pointing out areas for improvement. Finally, an overview of the state of research regarding the application of cardiac dECM and future challenges in bioengineering a human heart suitable for transplantation is provided.

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Perfusion via chemicals removes the DNA content of rabbit hearts while keeping the collagen and glycosaminoglycan unaltered

Seitzhaparova,

Kadyr,

Timur

et al. 2024

Preprint

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Despite new approaches in the treatment of cardiovascular disease (CVD) such as percutaneous coronary intervention, coronary artery bypass graft, and left ventricular assist devices, which cannot fully compensate for the effectiveness of the original heart, heart transplantation still remains as the most effective solution. A growing body of literature recognizes the importance of developing a whole heart constructed from living tissues to provide an alternative option for patients suffering from diseases of the cardiovascular system. A potential solution that shows a promise is to generate cell-free, i.e., decellularized, scaffolds using native heart tissue to be later cellularized and transplanted. This study reports the decellularization process and efficiency in an effort to create a whole heart scaffold. The hearts harvested from rabbits were perfused and the final bioartificial scaffolds were characterized for the efficiency of decellularization in terms of DNA content, collagen, and glycosaminoglycan. The compressive biomechanical properties of decellularized and native hearts were also determined and compared. Findings revealed that the DNA content of decellularized hearts was significantly reduced while keeping collagen and GAG content unchanged. Biomechanical properties of the hearth became inferior upon removal of the nuclear material. Decellularized hearts have significant importance in treating CVD as they serve as bioartificial hearts, providing a more clinically relevant model for potential human use. Future work will focus on the recellularization of the heart using induced pluripotent or embryonic stem cells to test its functionality.

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Development of a new decellularization protocol for the whole porcine heart

Cited by 5 publications

References 20 publications

Optimizing decellularization protocols for human thyroid tissues: a step towards tissue engineering and transplantation

Optimizing decellularization protocols for human thyroid tissues: a step towards tissue engineering and transplantation

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

Perfusion via chemicals removes the DNA content of rabbit hearts while keeping the collagen and glycosaminoglycan unaltered

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