Name: application of hemostatic devices in laparoscopic hepatectomy
Uploaded: 2022-04-19T13:00:00
Description: Watch this Scientific Journal Video about Application of Hemostatic Devices in Laparoscopic Hepatectomy at JoVE.com

Este trabalho foi apoiado por subsídios do Projeto de Ciência e Tecnologia da Cidade de Guangzhou (202102010090).

A operação é rotineira e recebeu aprovação ética. As aprovações são as seguintes: Aprovação para rápida revisão pelo Comitê de Pesquisa Clínica e Ética de Aplicação do Segundo Hospital Afiliado da Universidade Médica de Guangzhou: O projeto de revisão da ética da pesquisa científica "Aplicação de dispositivos hemostáticos simples e fáceis na Hepatectomia Laparoscópica" (aceitação Nº: 2022-hg-ks-02) foi aprovado pelo comitê de pesquisa clínica e ética de aplicação do Segundo Hospital Afi...

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Y., Han, H. S., Yoon, Y. S., Shin, S. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Feasibility+of+laparoscopic+liver+resection+for+tumors+located+in+the+posterosuperior+segments+of+the+liver,+with+a+special+reference+to+overcoming+current+limitations+on+tumor+location.">Feasibility of laparoscopic liver resection for tumors located in the posterosuperior segments of the liver, with a special reference to overcoming current limitations on tumor location.</a> Surgery. 144 (1), 32-38 (2008).</li><li>Ome, Y., Honda, G., Doi, M., Muto, J., Seyama, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laparoscopic+anatomic+liver+resection+of+segment+8+using+intrahepatic+Glissonean+approach.">Laparoscopic anatomic liver resection of segment 8 using intrahepatic Glissonean approach.</a> Journal of the American College of Surgeons. 230 (3), 13-20 (2020).</li><li>Ryu, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perioperative+and+oncological+outcomes+of+laparoscopic+anatomical+hepatectomy+for+hepatocellular+carcinoma+introduced+gradually+in+a+single+center.">Perioperative and oncological outcomes of laparoscopic anatomical hepatectomy for hepatocellular carcinoma introduced gradually in a single center.</a> Surgical Endoscopy. 32 (2), 790-798 (2018).</li><li>Makabe, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficacy+of+occlusion+of+hepatic+artery+and+risk+of+carbon+dioxide+gas+embolism+during+laparoscopic+hepatectomy+in+a+pig+model.">Efficacy of occlusion of hepatic artery and risk of carbon dioxide gas embolism during laparoscopic hepatectomy in a pig model.</a> Journal of Hepato-Biliary-Pancreatic Sciences. 21 (8), 592-598 (2014).</li><li>Miyagi, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pure+laparoscopic+hepatectomy+combined+with+a+pure+laparoscopic+pringle+maneuver+in+patients+with+severe+cirrhosis.">Pure laparoscopic hepatectomy combined with a pure laparoscopic pringle maneuver in patients with severe cirrhosis.</a> Case Reports in Gastroenterology. 9 (1), 101-105 (2015).</li></ol>

A hepatectomia é um dos métodos de tratamento mais importantes para pacientes com câncer de fígado primário e câncer de fígado secundário. Em 1991, Reich et al. aplicaram laparoscopia para remover tumores benignos do fígado e completaram a primeira hepatectomia laparoscópica4 do mundo. Após anos de rápido desenvolvimento e promoção, a hepatectomia laparoscópica tornou-se um dos principais métodos para tratar tumores malignos hepáticos e tem sido amplamente realizada nos últimos a...

O carcinoma hepatocelular é um dos tumores malignos mais comuns do sistema digestivo. De acordo com os diferentes locais primários, ele pode ser dividido em carcinoma hepatocelular primário e câncer de fígado secundário. As células podem metástase de tumores primários de órgãos fora do fígado para o fígado de várias maneiras, levando ao carcinoma no fígado. Foi relatado que mais de 50% das células metastáticas no fígado vêm de câncer colorretal, enquanto outras são de tumores de mama, pâncreas, pulmão e estômago, etc. Nos últimos anos, muitos tratamentos para câncer de fígado secundário estão disponíveis, incluindo quimioterapia sistemática, terapia intervencionista, terapia molecular direcionada, cirurgia, etc2. No entanto, a ressecção radical ainda é o tratamento mais eficaz porque pode remover completamente o câncer3.
Com o rápido desenvolvimento da tecnologia laparoscópica, a hepatectomia anatômica laparoscópica é gradualmente reconhecida pelos cirurgiões, mas ainda não é amplamente realizada, especialmente em instituições médicas primárias. Uma das razões é a exigência de dispositivos hemostáticos de alto grau. Estes são necessários para reduzir os riscos de sangramento e vazamento de bile durante o processo de operação. Aqui, apresentamos um conjunto de equipamentos hemostáticos simples e fáceis, incluindo um cateter de lúmen único, bisturi harmônico e eletrocoagulação monopole, para realização de hepatectomia laparoscópica. Para isso, primeiro a porta hepatis é ocluída intermitentemente usando um único cateter de lúmen. O tecido de parênquimo hepático é então ressecado usando o bisturi harmônico. Os pontos sanguíneos são coagulados por eletrocoagulação monopólio ponto a ponto. Este equipamento hemostático simples e fácil usa um cateter de lúmen único para realizar a manobra de pringle e utiliza a hemostasia do bisturi harmônico e a eletrocoagulação monopole. O equipamento pode ser encontrado facilmente em hospitais, proporcionando assim mais facilidade para a realização de laparoscopia ou treinamento. Assim, esses dispositivos hemostáticos simples e fáceis são adequados para a realização de procedimentos em instituições médicas primárias.
Neste estudo, o paciente foi um homem de 67 anos diagnosticado com um adenocarcinoma moderadamente diferenciado do cólon sigmoide metástase no fígado. A ressecção radical do câncer de cólon sigmoide foi realizada em janeiro de 2021. Os resultados patológicos foram adenocarcinoma moderadamente diferenciado com a etapa TNM do pT4aN2aM1. A quimioterapia FOLFOX foi dada quatro vezes após a operação. Na sequência, a condição corporal do paciente foi considerada adequada para realizar hepatectomia laparoscópica anatômica para remover completamente a lesão das metástases hepáticas. O grau child-pugh da função hepática era grau A. Para o teste de função de reserva hepática, R15 no teste de liberação do ICG foi de 1,6% (<10%). A tomografia mostrou um tumor de 57 mm x 68 mm x 76 mm na parte dorsal de S5 e parte ventral do S6 do fígado; o modelo de reconstrução tridimensional é mostrado na Figura 1.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>HARMONIC ACE Ultrasonic Surgical Devices</td>
			<td>Ethicon Endo-Surgery,LLC</td>
			<td>HAR36</td>
			<td></td>
		</tr>
		<tr>
			<td>Hem-o-lock</td>
			<td>Teleflex Medical</td>
			<td>544233</td>
			<td></td>
		</tr>
		<tr>
			<td>Monopole Electrocoagulation</td>
			<td>KANGJI MEDICAL</td>
			<td>/</td>
			<td></td>
		</tr>
		<tr>
			<td>single lumen ureter</td>
			<td>WELL LEAD MEDICAL CO.,LTD.</td>
			<td>12F</td>
			<td></td>
		</tr>
	</tbody>
</table>

application of hemostatic devices in laparoscopic hepatectomy

A hepatectomia laparoscópica é considerada um método convencional para o tratamento de doenças hepáticas benignas e malignas, por ser um método minimamente invasivo. Apesar de seu aspecto não invasivo, o sangramento e o vazamento de biliagem ocorrem na ressecção do tecido do parnchyma hepático durante a operação ou no período pós-operação, indicando a exigência de dispositivos hemostáticos de alto grau, como aspiração cirúrgica ultrassônica, eletrocoagulação bipolar, etc. A falta de disponibilidade desses dispositivos hemostáticos de alto grau impede que a hepatectomia laparoscópica se torne um procedimento generalizado em organizações médicas básicas. Tendo em vista a situação mencionada acima, um conjunto de dispositivos hemostáticos simples e fáceis é desenvolvido neste protocolo, que inclui um bisturi harmônico, eletrocoagulação monopole, e um único cateter de lúmen, para realizar inovadoramente a ressecção do tecido do parênco hepático. Em primeiro lugar, o porta hepatis ou pediculo hepático é ocluído intermitentemente por um único cateter de lúmen, seguido de fixação por 15 min e liberação por 5 min. Posteriormente, utilizando o bisturi harmônico, a fixação e o esmagamento do fígado são feitos para cortar o tecido de parenchyma hepático e revelar as artérias intrahpépticas, veias e ductos biliares. Por fim, os pontos sanguíneos são coagulados usando eletrocoagulação monopólio em cada ponto. As estruturas de tubulação intraháptica são então visíveis usando esses métodos, que poderiam parar de sangrar facilmente, reduzir a taxa de incidência de vazamento de bilia e melhorar a segurança e a viabilidade da hepatectomia laparoscópica. Portanto, os dispositivos hemostáticos simples e fáceis aqui mostrados são adequados para a realização de procedimentos em instituições médicas primárias.

A operação foi concluída em 2,5 h e o volume de sangramento intraoperatório foi de 100 mL sem a transfusão de sangue. Não houve complicações de curto prazo, e o paciente teve alta hospitalar no dia 8após a operação. O nível pós-operação do antígeno carcinoembrínico (CEA) diminuiu para 110,64 μg/L do nível de pré-operação de 1058,69 μg/L (Figura 12). Dois meses após a operação, o paciente retornou ao hospital para continuar o tratamento quimioterápico (o ...

Watch this Scientific Journal Video about Application of Hemostatic Devices in Laparoscopic Hepatectomy at JoVE.com

Application of Hemostatic Devices in Laparoscopic Hepatectomy

Dispositivos hemostáticos de alto grau são essenciais para hepatectomia laparoscópica. No entanto, esses dispositivos não são generalizados em organizações médicas básicas. Portanto, um conjunto de dispositivos hemostáticos simples e fáceis é mostrado neste artigo, o que pode tornar a hepatectomia laparoscópica mais fácil de realizar.

Aplicação de Dispositivos Hemostáticos em Hepatectomia Laparoscópica

Laparoscopic hepatectomy is considered a conventional method for treating benign and malignant liver diseases because it is a minimally invasive method. ...

A principal vantagem dessa técnica é que o dispositivo hemostático é simples e fácil de obter e pode ser realizado em hospitais básicos. Este conjunto de equipamentos hemostáticos pode ser aplicado ainda mais à hepatectomia laparoscópica e outros sistemas. É altamente recomendável ter uma certa base de cirurgia laparoscópica, especialmente o uso qualificado de um bisturi harmônico para aplicar este conjunto de dispositivos hemostáticos em hepatectomia laparoscópica.Comece aplicando punção direta para estabelecer pneumopertoneum na laparoscopia. Coloque um trocarte de 10 milímetros em cada orifício de observação na área subumbilical, bem no meio da linha clavicular sob a margem costal e a linha média ventral. Em seguida, coloque um trocarte de cinco milímetros na linha auxiliar anterior direita e na linha média ventral.Monte os dispositivos hemostáticos a partir do bisturi harmônico, eletrocoagulação monopólio e cateter de lúmen único para ser usado de forma inovadora na ressecção do tecido do parênquim hepático. Para realizar uma manobra de Pringle, oclui o primeiro porta hepatis intermitentemente usando um único cateter de lúmen. Aperte o cateter de lúmen com um clipe hemostático por 15 minutos seguido de soltá-lo por cinco minutos usando o bisturi harmônico para cortar o clipe e retirar o clipe para restaurar o suprimento de sangue para a primeira porta hepatis.Separe os pediculos hepáticos que fornecem o tumor. Ressecção e divida-os um a um com um bisturi harmônico. Observe a linha de isquemia na superfície hepática, que se torna visível após a ressecção dos pediculos hepáticos.Use o gancho para marcá-lo. Ressele o tecido de parenchyma hepático usando o bisturi harmônico enquanto aperta e esmaga o fígado gradualmente. Observe a artéria hepática interna, veia e ducto biliar.Resecte e divida essas estruturas de gasodutos. Uma vez que o sangramento apareça, coagula os pontos sanguíneos na superfície de dissecção hepática imediatamente usando a eletrocoagulação do monopólio ponto a ponto. Use o pediculo hepático posterior direito como uma placa de sinalização para ressecção de tecido parenchyma intrahpático.Reserve o principal pediculo hepático posterior e disseque o ramo de pedículos que abastece o tumor. Para completar a operação, lave a superfície do fígado para que se possa ver se ainda há manchas de sangramento. Pare o sangramento na superfície de dissecção do fígado e remova a amostra tumoral da cavidade abdominal.A operação foi concluída em 2,5 horas. O volume de sangramento intraoperatório foi de 100 mililitros sem transfusão de sangue e não foram observadas complicações de curto prazo. O nível de antígeno carcinomúnico pós-operação diminuiu para 110,64 microgramas por litro do nível de pré-operação de 1058,69 microgramas por litro.O tratamento quimioterápico foi retomado dois meses após a operação, de acordo com os resultados da patologia tumoral. O exame de tomografia pós-operatória foi comparado com a tomografia pré-operatória e não foram observados sinais de recrescimento tumoral. O passo mais importante no processo cirúrgico é o uso do bisturi harmônico.Uma boa quantidade de habilidade é necessária para evitar danos a estruturas vitais.

application-of-hemostatic-devices-in-laparoscopic-hepatectomy

Research

JoVE Journal

Cancer Research

Evaporation-reducing Culture Condition Increases the Reproducibility of Multicellular Spheroid Formation in Microtiter Plates

Tumor models that closely imitate in vivo conditions are becoming increasingly popular in drug discovery and development for the screening of potential anti-cancer drugs. Multicellular tumor spheroids (MCTSes) effectively mimic the physiological conditions of solid tumors, making them excellent in vitro models for lead optimization and target validation. Out of the various techniques available for MCTS culture, the liquid-overlay method on agarose is one of the most inexpensive methods for MCTS generation. However, the reliable transfer of MCTS cultures using liquid-overlay for high-throughput screening may be compromised by a number of limitations, including the coating of microtiter plates (MPs) with agarose and the irreproducibility of uniform MCTS formation across wells. MPs are significantly prone to edge effects that result from excessive evaporation of medium from the exterior of the plate, preventing the use of the entire plate for drug tests. This manuscript provides detailed technical improvements to the liquid-overlay technique to increase the scalability and reproducibility of uniform MCTS formation. Additionally, details on a simple, semi-automatic, and universally applicable software tool for the evaluation of MCTS features after drug treatment is presented.

The uneven loss of medium from microtiter plates affects the reproducibility of uniform multicellular tumor spheroid formation. Improving culture conditions to reduce significant medium loss will improve the reproducibility of spheroid formation and the results of spheroid-based assays using the liquid-overlay technique.

A evaporação de redução de Cultura condição aumenta a reprodutibilidade da Formação multicelular esferoidal em placas de microtitulação

Tumor models that closely imitate in vivo conditions are becoming increasingly popular in drug discovery and development for the screening of potential ...

Pesquisa do câncer

Application of Laser Micro-irradiation for Examination of Single and Double Strand Break Repair in Mammalian Cells

Highly coordinated DNA repair pathways exist to detect, excise and replace damaged DNA bases, and coordinate repair of DNA strand breaks. While molecular biology techniques have clarified structure, enzymatic functions, and kinetics of repair proteins, there is still a need to understand how repair is coordinated within the nucleus. Laser micro-irradiation offers a powerful tool for inducing DNA damage and monitoring the recruitment of repair proteins. Induction of DNA damage by laser micro-irradiation can occur with a range of wavelengths, and users can reliably induce single strand breaks, base lesions and double strand breaks with a range of doses. Here, laser micro-irradiation is used to examine repair of single and double strand breaks induced by two common confocal laser wavelengths, 355 nm and 405 nm. Further, proper characterization of the applied laser dose for inducing specific damage mixtures is described, so users can reproducibly perform laser micro-irradiation data acquisition and analysis.

Confocal fluorescence microscopy and laser micro-irradiation offer tools for inducing DNA damage and monitoring the response of DNA repair proteins in selected sub-nuclear areas. This technique has significantly advanced our knowledge of damage detection, signaling, and recruitment. This manuscript demonstrates these technologies to examine single and double strand break repair.

Aplicação de Laser Microirradiação para exame de Single e Double Strand reparação em células de mamíferos

Highly coordinated DNA repair pathways exist to detect, excise and replace damaged DNA bases, and coordinate repair of DNA strand breaks. While molecular ...

Laparoscopic Pancreatoduodenectomy With Modified Blumgart Pancreaticojejunostomy

Minimally invasive pancreatic resections are technically demanding but rapidly increasing in popularity. In contrast to laparoscopic distal pancreatectomy, laparoscopic pancreatoduodenectomy (LPD) has not yet obtained wide acceptance, probably due to technical challenges, especially regarding the pancreatic anastomosis.
The study describes and demonstrates all steps of LPD, including the modified Blumgart pancreaticojejunostomy. Indications for LPD are all pancreatic and peri-ampullary tumors without vascular involvement. Relative contra-indications are body mass index &#62;35 kg/m2, chronic pancreatitis, mid-cholangiocarcinomas and large duodenal cancers.
The patient is in French position, 6 trocars are placed, and dissection is performed using an (articulating) sealing device. A modified Blumgart end-to-side pancreaticojejunostomy is performed with 4 large needles (3/0) barbed trans-pancreatic sutures and 4 to 6 duct-to-mucosa sutures using 5/0 absorbable multifilament combined with a 12 cm, 6 or 8 Fr internal stent using 3D laparoscopy. Two surgical drains are placed alongside the pancreaticojejunostomy.
The described technique for LPD including a modified Blumgart pancreatico-jejunostomy is well standardized, and its merits are currently studied in the randomized controlled multicenter trial. This complex operation should be performed at high-volume centers where surgeons have extensive experience in both open pancreatic surgery and advanced laparoscopic gastro-intestinal surgery.

Laparoscopic pancreatoduodenctomy (LPD) may offer advantages over open pancreatoduodenectomy, including early postoperative mobilization, less delayed gastric emptying and a shorter hospital stay. However, LPD is technically challenging and not well-standardized, especially regarding the pancreatic anastomosis. We describe a standardized technique for the pancreatic anastomosis during LPD: modified Blumgart pancreaticojejunostomy.

Duodenopancreatectomia laparoscópica com modificado Blumgart pancreatoyeyunostomía

Minimally invasive pancreatic resections are technically demanding but rapidly increasing in popularity. In contrast to laparoscopic distal ...

The Application of 1% Methylene Blue Dye As a Single Technique in Breast Cancer Sentinel Node Biopsy

In this study, we injected 1% methylene blue dye (MBD) into the subareolar or peritumoral space of the breast. In the case of breast conserving surgery (BCS), a separate incision in the lower axilla hairline was made to find the sentinel nodes (SNs). In mastectomy, the SNs were identified through the same mastectomy incision. The SNs were described as blue nodes or nodes with lymphatic blue channels. An anatomical landmark in the axilla was used to facilitate SNs identification. The SNs metastases were evaluated by intraoperative frozen section analysis and histopathology examination as it is a gold standard. Here, we described the MBD as the lone technique in breast cancer sentinel node biopsy (SNB) which could be useful when radioisotope tracer or patent or isosulfan blue dye (PBD) cannot be provided.

In Indonesia, sentinel node biopsy (SNB) is not routinely performed for breast cancer surgery because of the limitation to provide radioisotope tracer and isosulfan or patent blue dye (PBD). To overcome this obstacle, we applied 1% methylene blue dye (MBD) as a single agent to map the sentinel nodes (SNs).

A aplicação de 1% de corante azul de metileno como uma única técnica na biópsia do nó Sentinela do cancro da mama

In this study, we injected 1% methylene blue dye (MBD) into the subareolar or peritumoral space of the breast. In the case of breast conserving surgery ...

The Clinical Application of Tumor Treating Fields Therapy in Glioblastoma

Glioblastoma is the most common and lethal form of brain cancer, with a median survival of 15 months after diagnosis and a 5 year survival rate of only 5% with current standard of care. Tumors often recur within 9 months following initial surgery, radiation and chemotherapy, at which point treatment options become limited. This highlights the pressing need for the development of better therapeutics to prolong survival and increase the quality of life for these patients.
Tumor Treating Fields (TTFields) therapy was developed to take advantage of the effect of low frequency alternating electrical fields on cells for cancer therapy. TTFields have been demonstrated to disrupt cells during mitosis and slow tumor growth. There is also growing evidence that they act through stimulating immune responses within exposed tumors. The advantages of TTFields therapy include its noninvasive approach and increased quality of life compared to other treatment modalities such as cytotoxic chemotherapies. The Food and Drug Administration approved TTFields therapy for the treatment of recurrent glioblastoma in 2011 and for newly diagnosed glioblastoma in 2015. We report on the effects of TTFields during mitosis, the results of electric fields modeling, and proper transducer array placement. Our protocol outlines the clinical application of TTFields on a patient post-surgery, using the second-generation device.

Glioblastoma is the most common and aggressive primary brain malignancy in adults, with most tumors recurring after initial treatment. Tumor Treating Fields (TTFields) therapy is the newest treatment modality for glioblastoma. Here, we describe the proper application of TTFields-transducer arrays on patients and discuss theory and aspects of treatment.

A aplicação clínica do Tumor, tratamento de terapia de campos em Glioblastoma

Glioblastoma is the most common and lethal form of brain cancer, with a median survival of 15 months after diagnosis and a 5 year survival rate of only 5% ...

In Vitro Evaluation of Oncogenic Transformation in Human Mammary Epithelial Cells

Tumorigenesis is a multi-step process in which cells acquire capabilities&#160;that allow their growth, survival, and dissemination under hostile conditions. Different tests seek to identify and quantify these hallmarks of cancerous cells; however, they often focus on a single aspect of cellular transformation and, in fact, multiple tests are required for their proper characterization. The purpose of this work is to provide researchers with a set of tools to assess cellular transformation in vitro from a broad perspective, thereby making it possible to draw sound conclusions.
A sustained proliferative signaling activation is the major feature of tumoral tissues and can be easily monitored under in vitro conditions by calculating the number of population doublings achieved over time. Besides, the growth of cells in 3D cultures allows their interaction with surrounding cells, resembling what occurs in vivo. This enables the evaluation of cellular aggregation and, together with immunofluorescent labeling of distinctive cellular markers, to obtain information on another relevant feature of tumoral transformation: the loss of proper organization. Another remarkable characteristic of transformed cells is their capacity to grow without attachment to other cells and to the extracellular matrix, which can be evaluated with the anchorage assay.
Detailed experimental procedures to evaluate cell growth rate, to perform immunofluorescent labeling of cell lineage markers in 3D cultures, and to test anchorage-independent cell growth in soft agar are provided. These methodologies are optimized for Breast Primary Epithelial Cells (BPEC) due to its relevance in breast cancer; however, procedures can be applied to other cell types after some adjustments.

This protocol provides experimental in vitro tools to evaluate the transformation of human mammary cells. Detailed steps to follow-up cell proliferation rate, anchorage-independent growth capacity, and distribution of cell lineages in 3D cultures with basement membrane matrix are described.

Avaliação In Vitro da Transformação Oncogênica em Células Epiteliais Mamárias Humanas

Tumorigenesis is a multi-step process in which cells acquire capabilities&#160;that allow their growth, survival, and dissemination under hostile ...

Co-culture of Glutamatergic Neurons and Pediatric High-Grade Glioma Cells Into Microfluidic Devices to Assess Electrical Interactions

Pediatric high-grade gliomas (pHGG) represent childhood and adolescent brain cancers that carry a rapid dismal prognosis. Since there is a need to overcome the resistance to current treatments and find a new way of cure, modeling the disease as close as possible in an in vitro setting to test new drugs and therapeutic procedures is highly demanding. Studying their fundamental pathobiological processes, including glutamatergic neuron hyperexcitability, will be a real advance in understanding interactions between the environmental brain and pHGG cells. Therefore, to recreate neurons/pHGG cell interactions, this work shows the development of a functional in vitro model co-culturing human-induced Pluripotent Stem (hiPS)-derived cortical glutamatergic neurons pHGG cells into compartmentalized microfluidic devices and a process to record their electrophysiological modifications. The first step was to differentiate and characterize human glutamatergic neurons. Secondly, the cells were cultured in microfluidic devices with pHGG derived cell lines. Brain microenvironment and neuronal activity were then included in this model to analyze the electrical impact of pHGG cells on these micro-environmental neurons. Electrophysiological recordings are coupled using multielectrode arrays (MEA) to these microfluidic devices to mimic physiological conditions and to record the electrical activity of the entire neural network. A significant increase in neuron excitability was underlined in the presence of tumor cells.

Recent works uncover the neuronal impact on high-grade pediatric glioma (pHGG) cells and their reciprocal interactions. The present work shows the development of an in vitro model co-culturing pHGG cells and glutamatergic neurons and recorded their electrophysiological interactions to mimic those interactivities.

Co-cultura de neurônios glutamatergicos e células glioma pediátricas de alto grau em dispositivos microfluidos para avaliar interações elétricas

Pediatric high-grade gliomas (pHGG) represent childhood and adolescent brain cancers that carry a rapid dismal prognosis. Since there is a need to ...

Flexible Organic Electronic Devices for Pulsed Electric Field Therapy of Glioblastoma

Glioblastoma is difficult to eradicate with standard oncology therapies due to its high degree of invasiveness. Bioelectric treatments based on pulsed electric fields (PEFs) are promising for the improvement of treatment efficiency. However, they rely on rigid electrodes that cause acute and chronic damage, especially in soft tissues such as the brain. In this work, flexible electronics were used to deliver PEFs to tumors and the biological response was evaluated with fluorescent microscopy. Interdigitated gold electrodes on a thin, transparent parylene-C substrate were coated with the conducting polymer PEDOT:PSS, resulting in a conformable and biocompatible device. The effects of PEFs on tumors and their microenvironment were examined using various biological models. First, monolayers of glioblastoma cells were cultured on top of the electrodes to investigate phenomena in vitro. As an intermediate step, an in ovo model was developed where engineered tumor spheroids were grafted in the embryonic membrane of a quail. Due to the absence of an immune system, this led to highly vascularized tumors. At this early stage of development, embryos have no immune system, and tumors are not recognized as foreign bodies. Thus, they can develop fast while developing their own vessels from the existing embryo vascular system, which represents a valuable 3D cancer model. Finally, flexible electrode delivery of PEFs was evaluated in a complete organism with a functional immune system, using a syngenic, orthograft (intracranial) mouse model. Tumor spheroids were grafted into the brain of transgenic multi-fluorescent mice prior to the implantation of flexible organic electrode devices. A sealed cranial window enabled multiphoton imaging of the tumor and its microenvironment during treatment with PEFs over a period of several weeks.

This work describes the development of flexible interdigitated electrodes for implementation in 3D brain tumor models, namely, in vitro culture, in ovo model, and in vivo murine model. The proposed method can be used to evaluate the effects of pulsed electric fields on tumors at different levels of complexity.

Dispositivos eletrônicos orgânicos flexíveis para terapia de campo elétrico pulsado de glioblastoma

Glioblastoma is difficult to eradicate with standard oncology therapies due to its high degree of invasiveness. Bioelectric treatments based on pulsed ...

Generation and Culturing of High-Grade Serous Ovarian Cancer Patient-Derived Organoids

Organoids are 3D dynamic tumor models that can be grown successfully from patient-derived ovarian tumor tissue, ascites, or pleural fluid and aid in the discovery of novel therapeutics and predictive biomarkers for ovarian cancer. These models recapitulate clonal heterogeneity, the tumor microenvironment, and cell-cell and cell-matrix interactions. Additionally, they have been shown to match the primary tumor morphologically, cytologically, immunohistochemically, and genetically. Thus, organoids facilitate research on tumor cells and the tumor microenvironment and are superior to cell lines. The present protocol describes distinct methods to generate patient-derived ovarian cancer organoids from patient tumors, ascites, and pleural fluid samples with a higher than 97% success rate. The patient samples are separated into cellular suspensions by both mechanical and enzymatic digestion. The cells are then plated utilizing a basement membrane extract (BME) and are supported with optimized growth media containing supplements specific to the culturing of high-grade serous ovarian cancer (HGSOC). After forming initial organoids, the PDOs can sustain long-term culture, including passaging for expansion for subsequent experiments.

Patient-derived organoids (PDO) are a three-dimensional (3D) culture that can mimic the tumor environment in vitro. In high-grade serous ovarian cancer, PDOs represent a model to study novel biomarkers and therapeutics.

Geração e Cultivo de Organoides Derivados de Pacientes com Câncer de Ovário Seroso de Alto Grau

Organoids are 3D dynamic tumor models that can be grown successfully from patient-derived ovarian tumor tissue, ascites, or pleural fluid and aid in the ...

Application of the En Bloc Concept Combined with Anatomic Resection in Laparoscopic Hepatectomy

Laparoscopic hepatectomy has been reported in many studies, and it is the mainstream method of liver resection. In some particular cases, such as when there are tumors adjacent to the cystic bed, surgeons cannot palpate the surgical margins through the laparoscopic approach, which leads to uncertainty about R0 resection. Conventionally, the gallbladder is resected first, and the hepatic lobes or segments are resected second. However, tumor tissues can be disseminated in the above cases. To address this issue, based on the recognition of the porta hepatis and intrahepatic anatomy, we propose a unique approach to hepatectomy combined with gallbladder resection by en bloc anatomic resection in situ. Firstly, after dissecting the cystic duct, without cutting the gallbladder primarily, the porta hepatis is pre-occluded by the single lumen ureter; secondly, the left hepatic pedicle is made free by the gap of the Laennec membrane and Hilar plate; thirdly, the assistant is asked to drag the fundus of the gallbladder, and the liver parenchyma tissue is resected using a harmonic scalpel along the ischemia line on the liver surface and intraoperative ultrasound. The whole middle hepatic vein (MHV) and its tributaries appear completely; lastly, the left hepatic vein (LHV) is disconnected, and the specimen is taken out from the abdominal cavity. The tumor, gallbladder, and other surrounding tissues are resected en bloc, which meets the tumor-free criterion, and a wide incisal margin and R0 resection are achieved. Therefore, the laparoscopic hepatectomy with the combination of the en bloc concept and anatomic resection is a safe, effective, and radical method with low postoperative recurrence and metastasis.

Numerous studies have demonstrated the advantages of anatomic resection. Nonetheless, whether anatomic resection can increase R0 resection rates remains controversial. Consequently, the present study describes an innovative procedure involving the en bloc concept combined with anatomic resection in laparoscopic hepatectomy, which can reduce postoperative recurrence and metastasis.

Aplicação do Conceito em Bloco Combinado com Ressecção Anatômica em Hepatectomia Laparoscópica

Laparoscopic hepatectomy has been reported in many studies, and it is the mainstream method of liver resection. In some particular cases, such as when ...