Effects of high voltage nanosecond electric pulses on eukaryotic cells (in vitro): A systematic review

Napotnik, Tina Batista; Reberšek, Matej; Vernier, P. Thomas; Mali, Barbara Černič; Miklavčič, Damijan

doi:10.1016/j.bioelechem.2016.02.011

Cited by 167 publications

(91 citation statements)

References 87 publications

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“…First, this study utilized bulk electroporation to deliver plasmids into the human NP cells and there can be limitations associated with this technique namely high voltage pulses to allow plasmids to permeate through the lipid bilayer . Outcomes include potential apoptosis, necrosis or cellular dysfunction (possibly senescence) . Our results demonstrated high cell viability up to 4 weeks and cellular morphology characteristic of the cell type.…”

Section: Ecm Synthesis and Degradationmentioning

confidence: 94%

Nonviral Transfection With Brachyury Reprograms Human Intervertebral Disc Cells to a Pro‐Anabolic Anti‐Catabolic/Inflammatory Phenotype: A Proof of Concept Study

Tang

Richards

Khan

et al. 2019

Journal Orthopaedic Research

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Intervertebral disc (IVD) degeneration is a major contributor to chronic low back pain and is characterized by decreases in cellularity and proteoglycan synthesis, upregulation of matrix degradation, and increases in pro‐inflammatory factors with neurovascular invasion. Current treatments fail to target the underlying pathology or promote tissue repair and approaches such as viral transfection raise safety concerns due to mutagenesis and unwarranted immune responses. To avoid such concerns, nonviral transfection is a viable method of gene delivery into the host cell while bypassing the caveats of viral delivery. Brachyury is expressed in the developing notochord and is associated with an immature healthy nucleus pulposus (NP). We hypothesize that Brachyury can reprogram degenerate NP cells to a healthy pro‐anabolic phenotype with increased proteoglycan content and decreased expression of catabolic, inflammatory, and neurovascular markers. NP cells obtained from human autopsy and surgical tissues were transfected with plasmids encoding for Brachyury or an empty vector control via bulk electroporation. Post transfection, cells were seeded in three‐dimensional agarose constructs cultured over 4 weeks and analyzed for viability, gene expression, and proteoglycan. Results demonstrated successful transfection of both autopsy and surgical NP cells. We observed long‐term Brachyury expression, significant increased expression of NP phenotypic markers FOXF1, KRT19, and chondrogenic marker SOX9 with decreases in inflammatory cytokines IL1‐β/IL6, NGF, and MMPs and significant increases in glycosaminoglycan accumulation. These results highlight nonviral transfection with developmental transcription factors, such as Brachyury, as a promising method to reprogram degenerate human disc cells toward a healthy NP phenotype. Clinical significance: This project proposes a novel translational approach for the treatment of intervertebral disc degeneration via direct reprogramming of diseased human patient‐derived IVD cells to a healthy phenotype. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:2389–2400, 2019

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Section: Ecm Synthesis and Degradationmentioning

confidence: 94%

Nonviral Transfection With Brachyury Reprograms Human Intervertebral Disc Cells to a Pro‐Anabolic Anti‐Catabolic/Inflammatory Phenotype: A Proof of Concept Study

Tang

Richards

Khan

et al. 2019

Journal Orthopaedic Research

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“…Only 44% reported a formal risk of bias assessment (Cochrane, GRADE, SYRCLE, Tool for Studies with Diverse Designs), one reported their own grading system [56]. Of the four reviews reporting Cochrane or SYRCLE tools it was notable that the majority of domains were rated as unclear, reiterating that the study designs are not based on randomisation, allocation concealment or blinding [57–60]. This finding agrees with our own assessments and indicates that basic research requires its own risk of bias tools.…”

Section: Resultsmentioning

confidence: 99%

“…75% reviews did not consider how valid the markers were. One review did consider the marker validity [66], three discussed outcome validity or quantification methods [57,62,63]. 69% of reviews did not consider the impact of imprecision; those that did consider any aspect of imprecision reported minimal details.…”

Section: Resultsmentioning

confidence: 99%

A systematic review of the asymmetric inheritance of cellular organelles in eukaryotes: A critique of basic science validity and imprecision

Collins

Ross²,

Lang³

2017

PLoS ONE

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We performed a systematic review to identify all original publications describing the asymmetric inheritance of cellular organelles in normal animal eukaryotic cells and to critique the validity and imprecision of the evidence. Searches were performed in Embase, MEDLINE and Pubmed up to November 2015. Screening of titles, abstracts and full papers was performed by two independent reviewers. Data extraction and validity were performed by one reviewer and checked by a second reviewer. Study quality was assessed using the SYRCLE risk of bias tool, for animal studies and by developing validity tools for the experimental model, organelle markers and imprecision. A narrative data synthesis was performed. We identified 31 studies (34 publications) of the asymmetric inheritance of organelles after mitotic or meiotic division. Studies for the asymmetric inheritance of centrosomes (n = 9); endosomes (n = 6), P granules (n = 4), the midbody (n = 3), mitochondria (n = 3), proteosomes (n = 2), spectrosomes (n = 2), cilia (n = 2) and endoplasmic reticulum (n = 2) were identified. Asymmetry was defined and quantified by variable methods. Assessment of the statistical reliability of the results indicated only two studies (7%) were judged to have low concern, the majority of studies (77%) were 'unclear' and five (16%) were judged to have 'high concerns'; the main reasons were low technical repeats (<10). Assessment of model validity indicated that the majority of studies (61%) were judged to be valid, ten studies (32%) were unclear and two studies (7%) were judged to have 'high concerns'; both described 'stem cells' without providing experimental evidence to confirm this (pluripotency and self-renewal). Assessment of marker validity indicated that no studies had low concern, most studies were unclear (96.5%), indicating there were insufficient details to judge if the markers were appropriate. One study had high concern for marker validity due to the contradictory results of two markers for the same organelle. For most studies the validity and imprecision of results could not be confirmed. In particular, data were limited due to a lack of reporting of interassay variability, sample size calculations, controls and functional validation of organelle markers. An evaluation of 16 systematic reviews containing cell assays found that only 50% reported adherence to PRISMA or ARRIVE reporting guidelines and 38% reported a formal risk of bias assessment. 44% of the reviews did not consider how relevant or valid the models were to the research question. 75% reviews did not consider how valid the markers were. 69% of reviews did not consider the impact of the statistical reliability of the results. Future systematic reviews in basic or preclinical research should ensure the rigorous reporting of the statistical reliability of the results in addition to the validity of the methods. Increased awareness of the importance of reporting guidelines and validation tools is needed for the scientific community.

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“…Retelj et al, 2013; PAULY and SCHWAN, 1959; Sundararajan, 2008). Moreover, it was also hypothesized that pulse frequencies above 100 kHz allow the applied electric field to reach the intracellular structures (Batista Napotnik et al, 2016; Gowrishankar and Weaver, 2003; Schoenbach et al, 2001). Thus, it was believed that electric pulse trains that have nano-second pulse widths and frequencies above 100 kHz allow electric perturbation of voltage sensitive intracellular processes of the endoplasmic reticulum, mitochondria, and the nucleus (Chopinet et al, 2013; Schoenbach et al, 2004).…”

Section: Discussionmentioning

confidence: 99%

Irreversible electroporation inhibits pro-cancer inflammatory signaling in triple negative breast cancer cells

Goswami

Coutermarsh-Ott

Morrison

et al. 2017

Bioelectrochemistry

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Low-level electric fields have been demonstrated to induce spatial re-distribution of cell membrane receptors when applied for minutes or hours. However, there is limited literature on the influence on cell signaling with short transient high-amplitude pulses typically used in irreversible electroporation (IRE) for cancer treatment. Moreover, literature on signaling pertaining to immune cell trafficking after IRE is conflicting. We hypothesized that pulse parameters (field strength and exposure time) influence cell signaling and subsequently impact immune-cell trafficking. This hypothesis was tested in-vitro on triple negative breast cancer cells treated with IRE, where the effects of pulse parameters on key cell signaling factors were investigated. Importantly, real time PCR mRNA measurements and ELISA protein analyses revealed that thymic stromal lymphopoietin (TSLP) signaling was down regulated by electric field strengths above a critical threshold, irrespective of exposure times spanning those typically used clinically. Comparison with other treatments (thermal shock, chemical poration, kinase inhibitors) revealed that IRE has a unique effect on TSLP. Because TSLP signaling has been demonstrated to drive pro-cancerous immune cell phenotypes in breast and pancreatic cancers, our finding motivates further investigation into the potential use of IRE for induction of an anti-tumor immune response in vivo.

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Effects of high voltage nanosecond electric pulses on eukaryotic cells (in vitro): A systematic review

Cited by 167 publications

References 87 publications

Nonviral Transfection With Brachyury Reprograms Human Intervertebral Disc Cells to a Pro‐Anabolic Anti‐Catabolic/Inflammatory Phenotype: A Proof of Concept Study

Nonviral Transfection With Brachyury Reprograms Human Intervertebral Disc Cells to a Pro‐Anabolic Anti‐Catabolic/Inflammatory Phenotype: A Proof of Concept Study

A systematic review of the asymmetric inheritance of cellular organelles in eukaryotes: A critique of basic science validity and imprecision

Irreversible electroporation inhibits pro-cancer inflammatory signaling in triple negative breast cancer cells

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