Towards the creation of decellularized organ constructs using irreversible electroporation and active mechanical perfusion

Sano, Michael B.; Neal, Robert E.; García, Paulo A.; Gerber, David A.; Robertson, John L.; Davalos, Rafael V.

doi:10.1186/1475-925x-9-83

Cited by 93 publications

(68 citation statements)

References 54 publications

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“…The lethal thresholds for U87 brain cancer cells found in this study were 463 V/cm and 505 V/cm in 3D and 2D models, respectively, when 100 × 100 μs monopolar IRE pulses were delivered. Previous studies found lethal thresholds in liver tissue between 300 and 640 V/cm using similar IRE protocols363738, indicating that both the 2D and 3D culture models recapitulate effects seen in mammalian tissue. Fourteen of sixteen protocols conducted in 2D and 3D conditions showed no statistically significant difference in lethal threshold indicating that the 3D model has a limited effect on the lethal threshold and cells cultured in a 2D monolayer are a relatively effective model for studying H-FIRE.…”

Section: Discussionmentioning

confidence: 72%

Asymmetric Waveforms Decrease Lethal Thresholds in High Frequency Irreversible Electroporation Therapies

Sano

Fan

2017

Sci Rep

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Irreversible electroporation (IRE) is a promising non-thermal treatment for inoperable tumors which uses short (50–100 μs) high voltage monopolar pulses to disrupt the membranes of cells within a well-defined volume. Challenges with IRE include complex treatment planning and the induction of intense muscle contractions. High frequency IRE (H-FIRE) uses bursts of ultrashort (0.25–5 μs) alternating polarity pulses to produce more predictable ablations and alleviate muscle contractions associated with IRE. However, H-FIRE generally ablates smaller volumes of tissue than IRE. This study shows that asymmetric H-FIRE waveforms can be used to create ablation volumes equivalent to standard IRE treatments. Lethal thresholds (LT) of 505 V/cm and 1316 V/cm were found for brain cancer cells when 100 μs IRE and 2 μs symmetric H-FIRE waveforms were used. In contrast, LT as low as 536 V/cm were found for 2 μs asymmetric H-FIRE waveforms. Reversible electroporation thresholds were 54% lower than LTs for symmetric waveforms and 33% lower for asymmetric waveforms indicating that waveform symmetry can be used to tune the relative sizes of reversible and irreversible ablation zones. Numerical simulations predicted that asymmetric H-FIRE waveforms are capable of producing ablation volumes which were 5.8–6.3x larger than symmetric H-FIRE waveforms indicating that in vivo investigation of asymmetric waveforms is warranted.

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Section: Discussionmentioning

confidence: 72%

Asymmetric Waveforms Decrease Lethal Thresholds in High Frequency Irreversible Electroporation Therapies

Sano

Fan

2017

Sci Rep

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“…However, one recent study has investigated the in vitro use of NTIRE combined with vascular perfusion to decellularize porcine liver. The results showed that perfusion with lactated ringer's solution facilitated removal of cellular debris resulting from electroporation, but also showed the limited effective geometric distribution of this approach [96]. The amount of cellular remnants retained within the tissue remains to be determined.…”

Section: Decellularization Agentsmentioning

confidence: 99%

An overview of tissue and whole organ decellularization processes

Crapo¹,

Gilbert²,

Badylak³

2011

Biomaterials

2,847

3,125

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Biologic scaffold materials composed of extracellular matrix (ECM) are typically derived by processes that involve decellularization of tissues or organs. Preservation of the complex composition and three-dimensional ultrastructure of the ECM is highly desirable but it is recognized that all methods of decellularization result in disruption of the architecture and potential loss of surface structure and composition. Physical methods and chemical and biologic agents are used in combination to lyse cells, followed by rinsing to remove cell remnants. Effective decellularization methodology is dictated by factors such as tissue density and organization, geometric and biologic properties desired for the end product, and the targeted clinical application. Tissue decellularization with preservation of ECM integrity and bioactivity can be optimized by making educated decisions regarding the agents and techniques utilized during processing. An overview of decellularization methods, their effect upon resulting ECM structure and composition, and recently described perfusion techniques for whole organ decellularization techniques are presented herein.

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“…In translational studies, large animals such as pigs and sheep are preferred [25,26,27,28,29,30,31,32]. The use of pigs as organ donors is discussed controversially because of the immunological barrier due to the expression of galactose-1,3-alpha-galactose in porcine organs which may lead to hyperacute rejection of the repopulated organ.…”

Section: Source Of Organs For Liver Engineeringmentioning

confidence: 99%

“…Other approaches concerning the optimization of the decellularization process were also tested such as the combination of electroporation with simultaneous perfusion [32]. The cryochemical method calls for freezing the freshly explanted liver in distilled water up to -80°C on the 1st day and slowly thawing on the 2nd day prior to starting perfusion.…”

Section: The Decellularization Processmentioning

confidence: 99%

Bioengineered Livers: A New Tool for Drug Testing and a Promising Solution to Meet the Growing Demand for Donor Organs

et al. 2016

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Background: Organ engineering is a new innovative strategy to cope with two problems: the need for physiological models for pharmacological research and donor organs for transplantation. A functional scaffold is generated from explanted organs by removing all cells (decellularization) by perfusing the organ with ionic or nonionic detergents via the vascular system. Subsequently the acellular scaffold is reseeded with organ-specific cells (repopulation) to generate a functional organ. Summary: This review gives an overview of the state of the art describing the decellularization process, the subsequent quality assessment, the repopulation techniques and the functional assessment. It emphasizes the use of scaffolds as matrix for culturing human liver cells for drug testing. Further, it highlights the techniques for transplanting these engineered scaffolds in allogeneic or xenogeneic animals in order to test their biocompatibility and use as organ grafts. Key Messages: The first issue is the so-called decellularization, which is best explored and resulted in a multitude of different protocols. The most promising approach seems to be the combination of pulsatile perfusion of the liver with Triton X-100 and SDS via hepatic artery and portal vein. Widely accepted parameters of quality control include the quantitative assessment of the DNA content and the visualization of eventually remaining nuclei confirmed by HE staining. Investigations regarding the composition of the extracellular matrix focused on histological determination of laminin, collagen, fibronectin and elastin and remained qualitatively. Repopulation is the second issue which is addressed. Selection of the most suitable cell type is a highly controversial topic. Currently, the highest potential is seen for progenitor and stem cells. Cells are infused into the scaffold and cultured under static conditions or in a bioreactor allowing dynamic perfusion of the scaffold. The quality of repopulation is mainly assessed by routine histology and basic functional assays. These promising results prompted to consider the use of a liver scaffold repopulated with human cells for pharmacological research. Transplantation of the (repopulated) scaffold is the third topic which is not yet widely addressed. Few studies report the heterotopic transplantation of repopulated liver tissue without vascular anastomosis. Even fewer studies deal with the heterotopic transplantation of a scaffold or a repopulated liver lobe. However, observation time was still limited to hours, and long-term graft survival has not been reported yet. These exciting results emphasize the potential of this new and promising strategy to create physiological models for pharmacological research and to generate liver grafts for the transplant community to treat organ failure. However, the scientific need for further development in the field of liver engineering is still tremendous.

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Towards the creation of decellularized organ constructs using irreversible electroporation and active mechanical perfusion

Cited by 93 publications

References 54 publications

Asymmetric Waveforms Decrease Lethal Thresholds in High Frequency Irreversible Electroporation Therapies

Asymmetric Waveforms Decrease Lethal Thresholds in High Frequency Irreversible Electroporation Therapies

An overview of tissue and whole organ decellularization processes

Bioengineered Livers: A New Tool for Drug Testing and a Promising Solution to Meet the Growing Demand for Donor Organs

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