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

Questo lavoro è stato sostenuto da sovvenzioni del Science and Technology Project della città di Guangzhou (202102010090).

L'operazione è di routine e ha ricevuto l'approvazione etica. Le approvazioni sono le seguenti: Approvazione per una rapida revisione da parte del Comitato etico per la ricerca clinica e l'applicazione del Secondo ospedale affiliato dell'Università di Medicina di Guangzhou: Il progetto di revisione etica della ricerca scientifica "Applicazione di dispositivi emostatici semplici e facili nell'epatectomia laparoscopica" (accettazione n.: 2022-hg-ks-02) è stato approvato dal comitato etico di ricerca clinica e applicazio...

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A., Briceno, J., Wakabayashi, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+short-term+benefits+of+laparoscopic+liver+resection:+9000+cases+and+climbing.">Comparative short-term benefits of laparoscopic liver resection: 9000 cases and climbing.</a> Annals of Surgery. 263 (4), 761-777 (2016).</li><li>Makuuchi, M., Hasegawa, H., Yamazaki, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasonically+guided+subsegmentectomy.">Ultrasonically guided subsegmentectomy.</a> Surgery, Gynecology, and Obstetrics. 161 (4), 346-350 (1985).</li><li>Agrawal, S., Belghiti, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oncologic+resection+for+malignant+tumors+of+the+liver.">Oncologic resection for malignant tumors of the liver.</a> Annals of Surgery. 253 (4), 656-665 (2011).</li><li>Wakabayashi, 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=Laparoscopic+repeat+liver+resection+after+open+liver+resection:+A+comparative+study+from+a+single-centre.">Laparoscopic repeat liver resection after open liver resection: A comparative study from a single-centre.</a> Journal of Minimal Access Surgery. 16 (1), 59-65 (2020).</li><li>Berardi, G., 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=Parenchymal+sparing+anatomical+liver+resections+with+full+laparoscopic+approach:+description+of+technique+and+short-term+results.">Parenchymal sparing anatomical liver resections with full laparoscopic approach: description of technique and short-term results.</a> Annals of Surgery. 273 (4), 785-791 (2021).</li><li>Cho, J. 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>

L'epatectomia è uno dei metodi di trattamento più importanti per i pazienti con carcinoma epatico primario e carcinoma epatico secondario. Nel 1991, Reich et al. hanno applicato laparoscopia per rimuovere i tumori benigni del fegato e hanno completato la prima epatectomia laparoscopicaal mondo 4. Dopo anni di rapido sviluppo e promozione, l'epatectomia laparoscopica è diventata uno dei metodi tradizionali per il trattamento dei tumori maligni del fegato ed è stata ampiamente effettuata negli u...

Il carcinoma epatocellulare è uno dei tumori maligni più comuni dell'apparato digerente. Secondo i diversi siti primari, può essere diviso in carcinoma epatocellulare primario e carcinoma epatico secondario. Le cellule possono metastatizzare dai tumori primari da organi al di fuori del fegato nel fegato in vari modi, portando al carcinoma nel fegato. È stato riportato che oltre il 50% delle cellule metastatiche nel fegato proviene dal cancro del colon-retto, mentre altri provengono da tumori al seno, al pancreas, ai polmoni e allo stomaco, ecc. 1. Negli ultimi anni, sono disponibili molti trattamenti per il cancro al fegato secondario, tra cui chemioterapia sistematica, terapia interventistica, terapia a bersaglio molecolare, chirurgia, ecc.2. Tuttavia, la resezione radicale è ancora il trattamento più efficace perché può rimuovere completamente il cancro3.
Con il rapido sviluppo della tecnologia laparoscopica, l'epatectomia anatomica laparoscopica viene gradualmente riconosciuta dai chirurghi ma non viene ancora ampiamente eseguita, specialmente nelle istituzioni mediche primarie. Uno dei motivi è il requisito di dispositivi emostatici di alta qualità. Questi sono necessari per ridurre i rischi di sanguinamento e perdite biliarie durante il processo operativo. Qui, presentiamo una serie di apparecchiature emostatiche semplici e facili, tra cui un catetere a singolo lume, un bisturi armonico e un'elettrocoagulazione monopolare, per l'esecuzione di epatectomia laparoscopica. Per fare questo, prima la porta epatite viene occlusa a intermittenza utilizzando un catetere a singolo lume. Il tessuto del parenchima epatico viene quindi resecato utilizzando il bisturi armonico. Le macchie sanguinanti sono coagulate mediante elettrocoagulazione monopolare punto a punto. Questa semplice e facile apparecchiatura emostatica utilizza un catetere a singolo lume per eseguire la manovra pringle e utilizza l'emostasi del bisturi armonico e l'elettrocoagulazione monopolare. L'attrezzatura può essere trovata facilmente negli ospedali, fornendo così una maggiore facilità per l'esecuzione di laparoscopia o formazione. Pertanto, questi dispositivi emostatici semplici e facili sono adatti per condurre procedure in istituzioni mediche primarie.
In questo studio, il paziente era un maschio di 67 anni con diagnosi di un adenocarcinoma moderatamente differenziato del colon sigmoide metastatizzato nel fegato. La resezione radicale del cancro del colon sigmoideo è stata eseguita nel gennaio 2021. I risultati patologici sono stati adenocarcinoma moderatamente differenziato con lo stadio TNM di pT4aN2aM1. La chemioterapia FOLFOX è stata somministrata quattro volte dopo l'operazione. Successivamente, le condizioni corporee del paziente sono state ritenute idonee per eseguire l'epatectomia anatomica laparoscopica per rimuovere completamente la lesione delle metastasi epatiche. Il grado child-pugh di funzionalità epatica era di grado A. Per il test di funzionalità della riserva epatica, R15 nel test di clearance ICG è stato dell'1,6% (<10%). La TC ha mostrato un tumore di 57 mm x 68 mm x 76 mm attraverso la parte dorsale di S5 e la parte ventrale di S6 del fegato; il modello di ricostruzione tridimensionale è illustrato nella Figura 1.

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	<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

L'epatectomia laparoscopica è considerata un metodo convenzionale per il trattamento di malattie epatiche benigne e maligne perché è un metodo minimamente invasivo. Nonostante il suo aspetto non invasivo, il sanguinamento e la perdita di bile si verificano nella resezione del tessuto del parenchima epatico durante l'operazione o nel periodo post-operatorio, indicando la necessità di dispositivi emostatici di alta qualità, come l'aspirazione chirurgica ad ultrasuoni, l'elettrocoagulazione bipolare, ecc. La mancanza di disponibilità di questi dispositivi emostatici di alta qualità impedisce all'epatectomia laparoscopica di diventare una procedura generalizzata nelle organizzazioni mediche di base. In considerazione della situazione sopra menzionata, in questo protocollo viene sviluppata una suite di dispositivi emostatici semplici e facili, che include un bisturi armonico, un'elettrocoagulazione monopolare e un catetere a singolo lume, per eseguire in modo innovativo la resezione del tessuto del parenchima epatico. Prima di tutto, la porta epatite o peduncolo epatico è occlusa a intermittenza da un singolo catetere lume, seguito da bloccaggio per 15 minuti e rilascio per 5 minuti. Successivamente, utilizzando il bisturi armonico, il serraggio e lo schiacciamento del fegato vengono eseguiti per tagliare il tessuto epatico del parenchima e per rivelare le arterie intraepatiche, le vene e i dotti biliari. Infine, le macchie sanguinanti vengono coagulate utilizzando l'elettrocoagulazione monopolare in ogni punto. Le strutture delle tubazioni intraepatiche sono quindi visibili utilizzando questi metodi, che potrebbero fermare facilmente il sanguinamento, ridurre il tasso di incidenza delle perdite biliari e migliorare la sicurezza e la fattibilità dell'epatectomia laparoscopica. Pertanto, i dispositivi emostatici semplici e facili mostrati qui sono adatti per condurre procedure in istituzioni mediche primarie.

L'operazione è stata completata entro 2,5 ore e il volume di sanguinamento intraoperatorio è stato di 100 ml senza la trasfusione di sangue. Non ci sono state complicazioni a breve termine e il paziente è stato dimesso dall'ospedale l'8° giorno dopo l'operazione. Il livello di antigene carcinoembrionale (CEA) dopo l'operazione è sceso a 110,64 μg/L dal livello pre-operatorio di 1058,69 μg/L (Figura 12). Due mesi dopo l'operazione, il paziente è tornato in ospedale per cont...

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

Application of Hemostatic Devices in Laparoscopic Hepatectomy

I dispositivi emostatici di alta qualità sono essenziali per l'epatectomia laparoscopica. Tuttavia, questi dispositivi non sono generalizzati nelle organizzazioni mediche di base. Pertanto, in questo articolo viene mostrata una suite di dispositivi emostatici semplici e facili, che possono rendere l'epatectomia laparoscopica più facile da eseguire.

Applicazione di dispositivi emostatici nell'epatectomia laparoscopica

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

Il vantaggio principale di questa tecnica è che il dispositivo emostatico è semplice e facile da ottenere e può essere eseguito negli ospedali di base. Questo set di apparecchiature emostatiche può essere ulteriormente applicato all'epatectomia laparoscopica e ad altri sistemi. Si raccomanda vivamente di avere una certa base di chirurgia laparoscopica, in particolare l'uso esperto di un bisturi armonico per applicare questo set di dispositivi emostatici nell'epatectomia laparoscopica.Inizia applicando la puntura diretta per stabilire lo pneumoperitoneo in laparoscopia. Posiziona un trocar di 10 millimetri in ogni foro di osservazione nell'area subombelicale, la linea clavicolare media destra sotto il margine costale e la linea mediana ventrale. Quindi posizionare un trocar di cinque millimetri nella linea ausiliaria anteriore destra e nella linea mediana ventrale.Assemblare i dispositivi emostatici a partire dal bisturi armonico, dall'elettrocoagulazione monopolare e dal catetere a singolo lume per essere utilizzati in modo innovativo nella resezione del tessuto epatico del parenchima. Per eseguire una manovra di Pringle, occludere la prima porta epatite in modo intermittente utilizzando un catetere a singolo lume. Blocca il catetere del lume con una clip emostatica per 15 minuti, quindi rilascialo per cinque minuti usando il bisturi armonico per tagliare la clip e togliere la clip per ripristinare l'afflusso di sangue alla prima porta epatite.Separare i peduncoli epatici che forniscono il tumore. Resecteli e dividili uno per uno con un bisturi armonico. Osservare la linea di ischemia sulla superficie del fegato, che diventa visibile dopo la resezione dei peduncoli epatici.Usa il gancio per contrassegnarlo. Resettare il tessuto del parenchima epatico usando il bisturi armonico mentre si blocca e si schiaccia gradualmente il fegato. Osservare l'arteria epatica interna, la vena e il dotto biliare.Resettare e dividere queste strutture di pipeline. Una volta che appare il sanguinamento, coagulare immediatamente le macchie sanguinanti sulla superficie di dissezione del fegato usando l'elettrocoagulazione monopolare punto a punto. Utilizzare il peduncolo epatico posteriore destro come segnale per la resezione del tessuto del parenchima intraepatico.Riservare il peduncolo epatico posteriore principale e sezionare il ramo di peduncoli che forniscono il tumore. Per completare l'operazione, lavare la superficie del fegato in modo da poter vedere se ci sono ancora macchie sanguinanti. Fermare il sanguinamento sulla superficie di dissezione del fegato e rimuovere il campione tumorale dalla cavità addominale.L'operazione è stata completata entro 2,5 ore. Il volume di sanguinamento intraoperatorio è stato di 100 millilitri senza la trasfusione di sangue e non sono state osservate complicazioni a breve termine. Il livello di antigene carcinoembrionale post-operatorio è sceso a 110,64 microgrammi per litro dal livello pre-operatorio di 1058,69 microgrammi per litro.Il trattamento chemioterapico è stato ripreso due mesi dopo l'operazione, secondo i risultati della patologia tumorale. L'esame TC postoperatorio è stato confrontato con la TC preoperatoria e non sono stati osservati segni di ricrescita tumorale. Il passo più importante nel processo chirurgico è l'uso del bisturi armonico.È necessaria una buona quantità di abilità per evitare danni alle strutture vitali.

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.

Evaporazione di riduzione Cultura Condizione Aumenta la riproducibilità dei multicellulare Spheroid Formazione in micropiastre

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

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Ricerca sul cancro

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.

Applicazione di Micro-irradiazione del Laser per l'esame di Single e Double Strand Break Repair in cellule di mammifero

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.

Pancreatoduodenectomy laparoscopica con modificate Blumgart Pancreaticojejunostomy

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).

L'applicazione dell'1% di colorazione blu di metilene come singola tecnica nel cancro al seno 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 ...

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.

L'applicazione clinica del tumore trattamento della terapia di campi nel 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.

Valutazione in vitro della trasformazione oncogenica nelle cellule epiteliali mammarie umane

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-coltura di neuroni glutamatergici e cellule di glioma pediatrico di alto grado in dispositivi microfluidici per valutare le interazioni elettriche

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.

Dispositivi elettronici organici flessibili per la terapia del campo elettrico pulsato del 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.

Generazione e coltura di organoidi di alto grado derivati da pazienti con carcinoma ovarico sieroso

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.

Applicazione del concetto di En Bloc combinato con la resezione anatomica nell'epatectomia laparoscopica

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