Inverted orientation improves decellularization of whole porcine hearts

Lee, Po-Feng; Chau, Eric; Cabello, Rafael Cisneros; Yeh, Alvin T.; Sampaio, Luiz C.; Gobin, Andrea S.; Taylor, Doris A.

doi:10.1016/j.actbio.2016.11.047

Cited by 50 publications

(42 citation statements)

References 44 publications

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“…Decellularization improves the ability of porcine valves to serve as heart valve substitutes . The evaluation of the decellularized valve as a heart valve substitute must consider the physical, chemical, and biological changes experienced by the valves during the decellularization process .…”

Section: Discussionmentioning

confidence: 99%

Structural assessments in decellularized extracellular matrix of porcine semilunar heart valves: Evaluation of cell niches

Roderjan

Noronha

Stimamiglio

et al. 2019

Xenotransplantation

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Tissue-engineered heart valves aim to reproduce the biological properties of natural valves with anatomically correct structure and physiological performance. The closest alternative to creating an ideal heart valve substitute is to use decellularized porcine heart valves, due to their anatomy and availability. However, the immunological barrier and the structural maintenance limit the long-term physiological performance of decellularized porcine heart valves. This study investigated the extracellular matrix (ECM) structure of aortic and pulmonary porcine valves decellularized by a low concentration sodium dodecyl sulfate (SDS)-based method in order to determine the ECM scaffold (ECMS) conditions related to remodeling potential. To assess the structures of the leaflets and conduits of the heart valves, ECM components and their organization were evaluated by histology, biochemical analysis (BC), scanning electron microscopy, multiphoton microscopy, tensile test, immunofluorescence labeling (IF), and Raman microspectroscopy used to draw a profile of the cell niches. Histology and multiphoton imaging of decellularized aortic and pulmonary leaflets and conduits revealed a collagen and elastin histoarchitecture with rearrangement, loosening fibers, and glycosaminoglycan depletion confirmed by biochemistry quantification.The potential cytotoxicity of SDS residues was eliminated after 10 wash cycles. The mechanical properties of the structure of the valve indicated a functional resistance of decellularized ECM. The IF demonstrated the presence of basement membrane, suggesting a potential structure for host cell attachment. The RM analysis showed evidence of molecular interactions, suggesting conservation of the chemical composition, particularly among the protein molecular structures. The structural analyses performed in the semilunar porcine heart valves demonstrate that decellularized ECMS has structural properties that support physiological performance and potential host tissue integration. In fact, decellularized leaflet scaffolds were prone to cell interaction after human adipose-derived stromal cell seeding and culturing. Further analysis of biocompatibility, particularly the ECM-cell interaction, can elucidate the remodeling process, in preserved decellularized heart valve scaffold.

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Section: Discussionmentioning

confidence: 99%

Structural assessments in decellularized extracellular matrix of porcine semilunar heart valves: Evaluation of cell niches

Roderjan

Noronha

Stimamiglio

et al. 2019

Xenotransplantation

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“…Enzymatic methods are often combined with chemical methods to allow for better cell removal before detergent implementation . Physical methods rely on using freeze‐thaw cycles, agitation, or application of pressure to damage cells and allow for cell removal, and are commonly used in conjunction with physical or enzymatic methods to improve decellularization . Regardless of method, complete decellularization is readily available for applications of dECM materials in cardiac tissue engineering …”

Section: Solid Decmmentioning

confidence: 99%

“…The system measured and controlled tissue pressure during decellularization to maintain pressure at physiological values, which reduced DNA content, but maintained collagen, elastin, and GAG composition, as well as compressive modulus compared to native tissue. Lee et al used an inverted orientation of decellularization to help maintain integrity of thinner areas of the heart while effectively removing cells from the thickest components . Compared to venting the heart apex during decellularization, the inversion method showed higher coronary perfusion efficiency, better cellular and DNA removal, higher collagen and elastin content in the aortic valve, and better shape retention.…”

Section: Solid Decmmentioning

confidence: 99%

Decellularized Extracellular Matrix Materials for Cardiac Repair and Regeneration

Bejleri

Davis

2019

Adv Healthcare Materials

151

105

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Decellularized extracellular matrix (dECM) is a promising biomaterial for repairing cardiovascular tissue, as dECM most effectively captures the complex array of proteins, glycosaminoglycans, proteoglycans, and many other matrix components that are found in native tissue, providing ideal cues for regeneration and repair of damaged myocardium. dECM can be used in a variety of forms, such as solid scaffolds that maintain native matrix structure, or as soluble materials that can form injectable hydrogels for tissue repair. dECM has found recent success in many regeneration and repair therapies, such as for musculoskeletal, neural, and liver tissues. This review focuses on dECM in the context of cardiovascular applications, with variations in tissue and species sourcing, and specifically discusses advances in solid and soluble dECM development, in vitro studies, in vivo implementation, and clinical translation.

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“…In response to a shift toward 3D cell culture, often carried out within bioreactors [5–8] or intricate cell-based microfluidic systems [9], there is a need to accurately measure viable cell numbers without disturbing or sacrificing the complex tissue under examination [10], which may be produced from scarce patient-specific cells [11–13] or patient-derived induced pluripotent stem cells [14]. In addition to providing information on cellular growth kinetics within these systems, noninvasive measurement of cell number is necessary to provide an accurate reference point for tissue function and phenotype.…”

Section: Introductionmentioning

confidence: 99%

“…In addition to providing information on cellular growth kinetics within these systems, noninvasive measurement of cell number is necessary to provide an accurate reference point for tissue function and phenotype. Rigorous, reproducible, and noninvasive cell counting measurements are fundamental tools needed to enable evaluation of cell-based products (matrices with cells [8]) and biofabricated tissues [7, 13, 15–23]. Measurement assurance of cell counting methods is an important element of process control to accurately reference cell and tissue behavior, measure tissue function on a per-cell basis, evaluate for batch-to-batch variability, and compare data across experiments and between laboratories [24, 25].…”

Section: Introductionmentioning

confidence: 99%

Essential design considerations for the resazurin reduction assay to noninvasively quantify cell expansion within perfused extracellular matrix scaffolds

et al. 2017

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Precise measurement of cellularity within bioartificial tissues and extracellular matrix (ECM) scaffolds is necessary to augment rigorous characterization of cellular behavior, as accurate benchmarking of tissue function to cell number allows for comparison of data across experiments and between laboratories. Resazurin, a soluble dye that is reduced to highly fluorescent resorufin in proportion to the metabolic activity of a cell population, is a valuable, noninvasive tool to measure cell number. We investigated experimental conditions in which resazurin reduction is a reliable indicator of cellularity within three-dimensional (3D) ECM scaffolds. Using three renal cell populations, we demonstrate that correlation of viable cell numbers with the rate of resorufin generation may deviate from linearity at higher cell densities, lower resazurin working volumes, or longer incubation times that all contribute to depleting the pool of resazurin. In conclusion, while the resazurin reduction assay provides a powerful, noninvasive readout of metrics enumerating cellularity and growth within ECM scaffolds, assay conditions may strongly influence its applicability for accurate quantification of cell number. The approach and methodological recommendations presented herein may be used as a guide for application-specific optimization of this assay to obtain rigorous and accurate measurement of cellular content in bioengineered tissues.

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Inverted orientation improves decellularization of whole porcine hearts

Cited by 50 publications

References 44 publications

Structural assessments in decellularized extracellular matrix of porcine semilunar heart valves: Evaluation of cell niches

Structural assessments in decellularized extracellular matrix of porcine semilunar heart valves: Evaluation of cell niches

Decellularized Extracellular Matrix Materials for Cardiac Repair and Regeneration

Essential design considerations for the resazurin reduction assay to noninvasively quantify cell expansion within perfused extracellular matrix scaffolds

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