Name: step by step: microsurgical training method combining two nonliving animal models
Uploaded: 2015-05-09T14:00:00.000+00:00
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The authors have no acknowledgements.

1. Preparazione: <ol><li> Ottenere un microscopio da tavolo stereo, con una variabile di 3,5 x di zoom 45x, 200 mm di distanza di lavoro, il 55 a 75 mm variabile distanza pupillare e un accessorio luce principale anello. Qualsiasi microscopio simile è adeguata, può essere impostato su un tavolino e non è estremamente costoso. </li><li> Raccogliere e preparare una serie di strumenti di microchirurgia con due pickup di microchirurgia, una forbice da dissezione, e un ago-guida; questo è il minimo richiesto, si deve tener conto del fatto che questi sono strumenti pratica, e strumenti di qualità elevata finali non sono necessari. </li><li> Utilizzare 9/0 monofilamento micro-sutura per tutte le pratiche. Comprare in commercio cosce di pollo o di ali e cibo colorante al supermercato locale. </li><li> A 20 G angio-catetere viene utilizzato per le prove di pervietà sul pollo arterie femorali. Per le arterie brachiale è necessario un più sottile 27 G catetere. Questi possono essere acquistati su internet in una farmacia locale ocon gli strumenti impostati. Comprare una siringa disponibile in commercio per iniettare il colorante alimentare. </li></ol> 2. macroscopiche tecniche di dissezione: <ol><li> L&#39;approccio ai vasi femorali è semplice. Per cosce di pollo, eseguire una tecnica di dissezione smussa appena mediale al diafisi femorale, separare il muscolo femorale dall&#39;osso ed esporre l&#39;arteria femorale, vena e nervi. Il nervo può essere ulteriormente sezionato in due fascicoli distinti per i diversi tipi di pratiche di riparazione dei nervi. </li><li> Per accedere brachiale pollo ali di pollo, incidere la pelle longitudinalmente con una forbice, dalla spalla fino alla punta dell&#39;ala. Osservare l&#39;arteria che corre tra la bicipiti e tricipiti brachiale. </li></ol> 3. regime di allenamento consigliato: <ol><li> Formazione con l&#39;arteria femorale. NOTA: Il primo passo per il regime di allenamento è quello di praticare con il pollo arteria femorale. </li></ol><ol><li> Posizionare il pezzo al microscopio e iniziare la dissezione microchirurgico. Liberare l&#39;arteria femorale dai tessuti sottostanti; e dividerlo orizzontalmente. Eseguire un adventicectomy. Utilizzare i micro-pickup e micro-forbici per tagliare qualsiasi adventicia sovrastante lume arterioso. Sciacquare il lume interno con acqua di rubinetto regolare. Annotare l&#39;ora di inizio </li><li> Eseguire un end to end anastomosi. <ol><li> Avviare sulla parete posteriore dell&#39;arteria sul lato opposto all&#39;operatore. Posizionare il primo punto; avviare sul lato sinistro se si lavora in orizzontale, sul lato inferiore se si lavora in verticale. Passare l&#39;ago dall&#39;esterno dell&#39;arteria verso il lume e quindi dal lume all&#39;esterno dell&#39;arteria. </li><li> Legare il punto, eseguire un doppio nodo, e poi due singoli tiri direzioni alternate. Completa l&#39;intera parete di fondo con la tecnica sopra descritta. </li><li> Iniziando dal lato destro se si lavora in senso orizzontale o il lato superiore, se siete working verticalmente, posizionare un singolo punto dall&#39;esterno verso il lume e quindi dal lume verso l&#39;esterno. Legare il nodo come descritto in precedenza. Completa la parete frontale ripetendo la procedura descritta in 3.1.2.2. </li><li> Annotare il tempo finale questo vi permetterà di tenere un registro di tempo, in modo da poter controllare la vostra progressione. NOTA: Gli autori preferiscono una tecnica a tratta fino; ma qualsiasi tipo di sutura microchirurgica può essere provato. E &#39;raccomandabile che il tirocinante compie circa 25 anastomosi microchirurgiche in questo modo, prima di poter provare altri scenari clinici. Questi includono innesto vena di lacune arteriose, capo all&#39;altro riparazioni nervose, e l&#39;uso di condotti vena per il trattamento delle lacune nervose. Una tecnica importante nella chirurgia microvascolare, che può essere appresa con questo metodo di allenamento è la sutura continua o in esecuzione, per eseguire questa tecnica i proventi tirocinante come 3.1.2.1 ma il stich non viene tagliato, continuerà a svolgere punti come in 3.1.2.1, senza taglio li uino a completare l&#39;intera sutura e legarlo al punto originale alla fine. </li></ol></li></ol><ol start="2"><li> Allenamento con l&#39;arteria brachiale <ol><li> Posizionare il pezzo al microscopio. Completare la procedura descritta sopra per l&#39;arteria femorale. NOTA: L&#39;unica differenza è la dimensione. L&#39;arteria brachiale è molto più piccola l&#39;arteria femorale e l&#39;anastomosi è considerevolmente più difficile. </li></ol></li><li> Test di pervietà <ol><li> Attraversare un 20 o 27 G angio-catetere liscio attraverso l&#39;anastomosi, per verificare la presenza di ostruzioni causate da punti di sutura entrambe le pareti dei vasi insieme. </li><li> Applicare una siringa con commerciale colorante alimentare al fine della angio-catetere, e profumato tingere nell&#39;arteria per verificare l&#39;assenza di perdite jet </li><li> Aprire l&#39;arteria longitudinalmente di visualizzare i punti, il loro parallelismo e la quantità di parete presa. </li></ol></li></ol>

<ol>
	<li>Lee, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Historical+events+on+development+of+experimental+microsurgical+organ+transplantation.">Historical events on development of experimental microsurgical organ transplantation.</a> Yonsei Med J. 45 (6), 1115-1120 (2004).</li><li>Shurey, S., Akelina, Y., Legagneux, J., Malzone, G., Jiga, L., Ghanem, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+rat+model+in+microsurgery+education:+classical+exercises+and+new+horizons.">The rat model in microsurgery education: classical exercises and new horizons.</a> Arch Plast Surg. 41 (3), 201-208 (2014).</li><li>Olabe, J., Olabe, J., Sancho, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+cadaveric+infusi&#243;n+model+for+neurosurgical+training.">Human cadaveric infusi&#243;n model for neurosurgical training.</a> Surg Neurol. 72, 700-702 (2009).</li><li>Peled, I. J., Kaplan, H. Y., Wexler, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microsilicone+anastomoses.">Microsilicone anastomoses.</a> Ann Plast Surg. 10 (4), 331-332 (1983).</li><li>Gluer, N. M., Rao, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Canniesburn+&#8220;ever-ready&#8221;+model+to+practise+microsurgery.">Canniesburn &#8220;ever-ready&#8221; model to practise microsurgery.</a> Br J Plast Surg. 43 (3), 381-382 (1990).</li><li>Hino, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Training+in+microvascular+surgery+using+a+Chicken+wing+artery.">Training in microvascular surgery using a Chicken wing artery.</a> Neurosurgery. 52 (6), 1495-1498 (2003).</li><li>Marsh, D. J., Norton, S. E., Mok, J., Patel, H. D., Chen, H. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microsurgical+training+the+Chicken+thigh+model.">Microsurgical training the Chicken thigh model.</a> Ann Plast Surg. 59 (3), 355-356 (2007).</li><li>Jeong, H. S., Moon, M. S., Kim, H. S., Lee, H. K., Yi, S. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microsurgical+training+with+fresh+chicken+legs+and+their+histological+characteristics.">Microsurgical training with fresh chicken legs and their histological characteristics.</a> Ann Plast Surg. 70 (1), 57-61 (2013).</li><li>Couceiro, J., Ozyurekoglu, T., Sanders, S., Tien, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microsurgical+training+regimen+with+nonliving+chicken+models.">Microsurgical training regimen with nonliving chicken models.</a> Microsurgery. 33 (3), 251-252 (2013).</li><li>Phoon, A. F., Gumley, G. J., Rtshiladze, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microsurgical+training+using+a+pulsatile+membrane+pump+and+chicken+thigh:+A+new,+realistic,+practical,+nonliving+educational+model.">Microsurgical training using a pulsatile membrane pump and chicken thigh: A new, realistic, practical, nonliving educational model.</a> Plast Reconstr Surg. 126 (5), 278e-279e (2010).</li><li>Zhou, Y., Gu, X., Xiang, J., Qian, S., Chen, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+comparative+study+on+suture+versus+cuff+anastomosis+in+mouse+cervical+cardiac+transplant.">A comparative study on suture versus cuff anastomosis in mouse cervical cardiac transplant.</a> Exp Clin Transplant. 8 (3), 245-249 (2010).</li></ol>

The authors have nothing to disclose.

The clinical practice of microsurgery requires a continuous practice and a high level of expertise; it&#8217;s applications include free flaps, replantation and revascularization procedures, intracranial &#8211; extracranial anastomoses for neurosurgical procedures, peripheral nerve surgery and others. In spite of the wide variety of procedures that require microsurgical techniques, some of them are seldom performed in many centers. The surgeons therefore need sometimes to maintain their skill level rehearsing the surgeries on different practice models. 
The most widely spread training model is currently the rat 1,2; it is considered by many to be the &#8220;gold standard&#8221; in microsurgical instruction. Rats are indeed a very valuable tool in the teaching and learning of microsurgery. They provide a very similar environment to that of the clinical situation. This approach however exhibits some intrinsic limitations. The use of living animals presents an ethical dilemma; and is generally subjected to institutional review board approval. Furthermore they require anesthesia, and dedicated technicians and facilities. All These factors imply important economic costs; this restricts training on living animals to specific teaching units, experimental surgery departments and hospitals with similar resources. 
The use of nonliving animal models described in this protocol, has the advantage of not raising any ethical concerns; it does not require institutional review board approval. Economically it compares favorably to living animal training. The required equipment is limited and can be acquired over the internet for a reasonable price; the author&#8217;s tabletop stereomicroscope was bought for around 450 United States Dollars (USD) and the set of practice microsurgical instruments was purchased for around 150 USD. The author uses microsurgical sutures provided by his sponsoring institution; the price of sutures varies greatly and high quality sutures are not needed, a box of twelve sutures can be bought for around 70 USD on the internet. A microsurgical training station can be set up on a very limited area at home or at work; Jeong et al.8 Placed their microscope at the residents&#8217; office; whilst we9 used an available microscope close to the hand surgery fellows&#8217; quarters. 
The most critical point in the application of this protocol is the need for the trainee to be constant and to stay motivated with his training, it is therefore very important that he starts training with the bigger vessels first and then proceed to the smaller chicken wing vessels to avoid frustration. The limitations of training with nonliving models involve primarily the lack of bleeding and the inexistence of hemodynamic phenomena like vasospasm and clotting. The advantages and disadvantages of this type of training are summarized on Table 1. The three tests detailed in section three are aimed to help the trainee check the vessel&#8217;s patency in the absence of the earlier mentioned phenomena; they provide information on the quality of the trainee&#8217;s suture, leaking, suture misalignment, and lumen obstruction caused by stitches suturing the front and back wall together. 
The protocol can potentially be improved by the addition of a peristaltic pump10 to simulate vessel flow or by flooding the field with liquid to simulate superficial tension. It is the authors opinion that this would result in the production of an even more realistic training environment and this should be the direction of future research. 
Another possible field for development is the application of this training method to help the trainee learn more complex procedures, such as the cuff anastomosis technique11 before it&#8217;s actual application on animal research. The authors have however no experience with such techniques at present time. 
In conclusion the protocol described in the present manuscript allows a determined trainee to setup a small practice station and use it to learn, perfect and maintain microsurgical skills progressively and sustainably; without the need for special facilities and minimizing animal deaths.

An ample array of training models have been described for the learning and maintenance of both basic and advanced microsurgical skills. These include living animals1,2, human cadaveric specimens3, inert models4,5 and non-living animal models6-9.The Living animal models, specifically those using rats have been extensively used in the teaching of microsurgery 1,2; and they are considered the current gold standard of training. In spite of their remarkable instructive value, when an intensive or prolonged training is required; economical and ethical concerns can hinder their practical usage. 
Cadaveric models3 offer the opportunity to practice in an environment similar to that of the actual clinical situation; they are unfortunately restricted to anatomy labs and similar facilities, a microscope must be available at the lab as well; these models are therefore not widely available. Inert models such as rubber tubes, or gloves4,5 are cheap, and easily accessible, the required equipment for their application is minimal. The resemblance to a clinical situation is tenuous however; and their application is generally limited to the introductory stages of training, before the trainee starts practicing on rats. To the best of our knowledge Hino6 was the first to propose the use of non-living chicken for the development and maintenance of clinical microsurgical skills. His model is based on the extraction and subsequent usage of the chicken brachial artery from commercial chicken wings. This approach does not need institution review board approval, rodent anaesthesia, laboratory technicians or postoperative care. 
Following Hino&#180;s description of the chicken wing artery model numerous other nonliving animal models were published. The use of commercial chicken thighs was first suggested by Marsh et al.7 who reported the use of the femoral artery, vein and nerve for didactic purposes. This particular model expands the possibilities of training including structures different from the arteries; it permits the trainee to perform a variety of complex procedures such as vein grafts, nerve grafts or fascicular repairs. Jeong et al.8 Further studied Marsh&#8217; chicken thigh model and its applications to the teaching of resident doctors. The authors analysed chicken tissue samples histologically and found that it presented similarities to human tissues. The time to perform an anastomosis by the trainee was significantly reduced by practice in their study. They concluded that &#8220;the femoral neurovascular bundles of chicken are an appropriate and effective model for teaching and practicing microsurgery&#8221;. In 2013 my colleagues and I9 published a microsurgical training regimen that combined both the chicken thigh and the chicken wing models; this approach benefits from the versatility of the chicken thighs and from the limited size of the chicken wing artery which can represent more of a challenge than the femoral vessels for the more advanced trainees. 
On the current manuscript we describe step by step the process of setting up a microsurgical practice station; the materials required, how to dissect the neurovascular structures of each model and how to effectively apply the training regimen.

<table><tbody><tr><td>Stereomicroscope</td><td>Amscope</td><td>SM-1BX</td><td>Microscope</td></tr><tr><td>Led light</td><td>Amscope</td><td>LED-80S-YA</td><td>illumination system</td></tr><tr><td>Microsurgical instruments set</td><td>Chen hen microsurgical instruments</td><td>HC- X803-1</td><td>Microsurgical instruments</td></tr><tr><td>Microsurgical sutures</td><td>S/T microsurgical</td><td>9/0 Suture</td><td>Sutures</td></tr></tbody></table>

step by step: microsurgical training method combining two nonliving animal models

The learning of microsurgical techniques and the maintenance of microsurgical skills have been traditionally based on the use of living animals, mainly laboratory rats. This method although extremely valuable can be economically demanding both for the surgeon and the sponsoring institution; it also requires special training facilities that may not always be available or accessible. Furthermore ethical concerns can limit the use of living animals for training purposes. Alternative training methods, such as inert tubes and gloves have not gained popularity among surgeons since they do not offer an experience similar to that of a clinical situation. Non-living animal models include the use of chicken thighs and wings; they offer a practice experience that resembles a clinical situation to a considerable extent. This type of training is relatively cheap and easily available. The microscope and instruments required can be acquired over the internet, and the chicken pieces can be bought at the local supermarket.
This approach allows a motivated trainee to rehearse different types of surgical techniques several times at a reasonable expense, helping to develop or maintain his surgical expertise if more complex facilities are not available. On the current manuscript we describe how to setup a small practice station, how to dissect the specimens, and how to practice both with the chicken thighs and with the chicken wings in a progressive fashion. This approach takes advantage on the versatility of the chicken thigh model and the small size of the chicken wing Brachial artery.

Gli autori hanno chronometered loro tempi con il presente metodo di formazione e hanno visto ridurre sensibilmente i tempi per una anastomosi dell&#39;arteria femorale alla fine della terza settimana. Il tirocinante può controllare i miglioramenti sulla sua tecnica controllando i suoi tempi, le sue percentuali di pervietà e l&#39;allineamento dei suoi punti di sutura. Il sistema descritto in questo manoscritto è illustrato nella figura 1 comprende un microscopio da tavolo e una serie di strumenti di microchirurgia. Le dissezioni macroscopiche sono mostrati nelle Figure 2 e 3. Un esempio di un&#39;arteria femorale anastomosi microchirurgica è illustrato nella figura 4. La figura 5 mostra una arteria brachiale anastomosi microchirurgica. La figura 6 mostra una riparazione arteria femorale con un innesto venoso. I test di pervietà sono mostrati in Figura 7. <img alt="Figura 1" src="/files/ftp_upload/52625/52625fig1.jpg" /> Figura 1: La corrispondendo propria stazione pratica microchirurgico dell&#39;autore. Esso comprende un microscopio da tavolo e una serie di strumenti di microchirurgia formazione. Può essere installato su uno spazio molto limitato in ufficio oa casa. <img alt="Figura 2" src="/files/ftp_upload/52625/52625fig2.jpg" /> Figura 2: Una dissezione macroscopica di una coscia di pollo, i vasi femorali e nervo sciatico può essere facilmente esposto l&#39;esecuzione di un dito appena mediale al diafisi femorale. La freccia orizzontale indica la testa femorale. Il piano di dissezione è contrassegnato dalla freccia verticale parallelo al piano femorale all&#39;albero B: L&#39;arteria femorale (a) vena (v) e del nervo sciatico (n) sono mostrate in dettaglio qui. <img alt="Figura 3" src="/files/ftp_upload/52625/52625fig3.jpg" /> Figura 3 A e B: dissezione macroscopica di un&#39;ala di pollo. Il brachiale un<html> rtery corre tra la bicipiti e tricipiti brachiale. L&#39;arteria è contrassegnato da una freccia su entrambe le fotografie è abbastanza superficiale e l&#39;approccio è semplice; esso può essere realizzato con le forbici cottura commerciali. <img alt="Figura 4" src="/files/ftp_upload/52625/52625fig4.jpg" /> Figura 4: immagine al microscopio di un fine femorale alla fine di sutura, è stato eseguito qui un modo uno fino tecnica con i singoli punti di sutura. Qualsiasi altro tipo di sutura può essere praticato a discrezione del tirocinante. <img alt="Figura 5" src="/files/ftp_upload/52625/52625fig5.jpg" /> Figura 5: Fine per terminare la sutura di una arteria brachiale pollo. Questo arterie sono molto più piccole di quelle femorali e la procedura è molto più tecnicamente esigenti. Carico / 52625 / 52625fig6.jpg &quot;/&gt; Figura 6: Una riparazione arteria femorale con un innesto vena raccolto dalla vena femorale. Il modello di coscia di pollo è molto versatile consente per i diversi tipi di esercizi pratici. <img alt="Figura 7" src="/files/ftp_upload/52625/52625fig7.jpg" /> Figura 7:. I test di pervietà R: Un angio-catetere di diametro appropriato è attraversato attraverso l&#39;anastomosi per verificare la presenza di ostruzioni causate da stiches sutura sia delle pareti della nave. B:. Dye è perfuso attraverso l&#39;anastomosi per verificare l&#39;assenza di perdite C : L&#39;anastomosi è aperto longitudinalmente per visualizzare l&#39;allineamento di sutura.

Watch this Scientific Journal Video about Step By Step: Microsurgical training method combining two nonliving animal models at JoVE.com

Step By Step: Microsurgical training method combining two nonliving animal models

Passo dopo passo: metodo di allenamento Microsurgical combinare due modelli animali non viventi

Here we describe how to set up a small microsurgical practice station and a simple and inexpensive method for the training of microsurgery with non-living animal models.

The learning of microsurgical techniques and the maintenance of microsurgical skills have been traditionally based on the use of living animals, mainly ...

step-step-microsurgical-training-method-combining-two-nonliving

Universidad San Sebastian

Research

JoVE Journal

Medicine

15.2K Views.  POVISA Hospital. Here we describe how to set up a small microsurgical practice station and a simple and inexpensive method for the training of microsurgery with non-living animal models. 

Video: Step By Step: Microsurgical training method combining two nonliving animal models

A Novel Microsurgical Model for Heterotopic, En Bloc Chest Wall, Thymus, and Heart Transplantation in Mice

Exploration of novel strategies in organ transplantation to prolong allograft survival and minimizing the need for long-term maintenance immunosuppression must be pursued. Employing vascularized bone marrow transplantation and co-transplantation of the thymus have shown promise in this regard in various animal models.1-11 Vascularized bone marrow transplantation allows for the uninterrupted transfer of donor bone marrow cells within the preserved donor microenvironment, and the incorporation of thymus tissue with vascularized bone marrow transplantation has shown to increase T-cell chimerism ultimately playing a supportive role in the induction of immune regulation. The combination of solid organ and vascularized composite allotransplantation can uniquely combine these strategies in the form of a novel transplant model. Murine models serve as an excellent paradigm to explore the mechanisms of acute and chronic rejection, chimerism, and tolerance induction, thus providing the foundation to propagate superior allograft survival strategies for larger animal models and future clinical application. Herein, we developed a novel heterotopic en bloc chest wall, thymus, and heart transplant model in mice using a cervical non-suture cuff technique. The experience in syngeneic and allogeneic transplant settings is described for future broader immunological investigations via an instructional manuscript and video supplement.

To study combined solid organ and vascularized composite allotransplantation, we describe a novel heterotopic en bloc chest wall, thymus, and heart transplant model in mice using a cervical non-suture cuff technique.

A Novel microchirurgica modello per eterotopica, in blocco parete toracica, Timo, e trapianto cardiaco nei topi

Exploration of novel strategies in organ transplantation to prolong allograft survival and minimizing the need for long-term maintenance immunosuppression ...

Ricerca

Medicina

Preparation of Murine Submandibular Salivary Gland for Upright Intravital Microscopy

The submandibular salivary gland (SMG) is one of the three major salivary glands, and is of interest for many different fields of biological research, including cell biology, oncology, dentistry, and immunology. The SMG is an exocrine gland comprised of secretory epithelial cells, myofibroblasts, endothelial cells, nerves, and extracellular matrix. Dynamic cellular processes in the rat and mouse SMG have previously been imaged, mostly using inverted multi-photon microscope systems. Here, we describe a straightforward protocol for the surgical preparation and stabilization of the murine SMG in anesthetized mice for in vivo imaging with upright multi-photon microscope systems. We present representative intravital image sets of endogenous and adoptively transferred fluorescent cells, including the labeling of blood vessels or salivary ducts and second harmonic generation to visualize fibrillar collagen. In sum, our protocol allows for surgical preparation of mouse salivary glands in upright microscopy systems, which are commonly used for intravital imaging in the field of immunology.

We describe a protocol to surgically expose and stabilize the murine submandibular salivary gland for intravital imaging using upright intravital microscopy. This protocol is easily adaptable to other exocrine glands of the head and neck region of mice and other small rodents.

Preparazione della ghiandola salivare sottomandibolare murino per microscopia Intravital verticale

The submandibular salivary gland (SMG) is one of the three major salivary glands, and is of interest for many different fields of biological research, ...

Immunologia e infezioni

Surgical Training for the Implantation of Neocortical Microelectrode Arrays Using a Formaldehyde-fixed Human Cadaver Model

This protocol describes a procedure to assist surgeons in training for the implantation of microelectrode arrays into the neocortex of the human brain. Recent technological progress has enabled the fabrication of microelectrode arrays that allow recording the activity of multiple individual neurons in the neocortex of the human brain. These arrays have the potential to bring unique insight onto the neuronal correlates of cerebral function in health and disease. Furthermore, the identification and decoding of volitional neuronal activity opens the possibility to establish brain-computer interfaces, and thus might help restore lost neurological functions. The implantation of neocortical microelectrode arrays is an invasive procedure requiring a supra-centimetric craniotomy and the exposure of the cortical surface; thus, the procedure must be performed by an adequately trained neurosurgeon. In order to provide an opportunity for surgical training, we designed a procedure based on a human cadaver model. The use of a formaldehyde-fixed human cadaver bypasses the practical, ethical and financial difficulties of surgical practice on animals (especially non-human primates) while preserving the macroscopic structure of the head, skull, meninges and cerebral surface and allowing realistic, operating room-like positioning and instrumentation. Furthermore, the use of a human cadaver is closer to clinical daily practice than any non-human model. The major drawbacks of the cadaveric simulation are the absence of cerebral pulsation and of blood and cerebrospinal fluid circulation. We suggest that a formaldehyde-fixed human cadaver model is an adequate, practical and cost-effective approach to ensure proper surgical training before implanting microelectrode arrays in the living human neocortex.

We designed a procedure in which a formaldehyde-fixed human cadaver is used to assist neurosurgeons in training for the implantation of microelectrode arrays into the neocortex of the human brain.

Training chirurgico per l'impianto di microelettrodi Neocortical utilizzando un modello di cadavere umano formaldeide-fisso

This protocol describes a procedure to assist surgeons in training for the implantation of microelectrode arrays into the neocortex of the human brain. ...

Porcine As a Training Module for Head and Neck Microvascular Reconstruction

Live models that resemble surgical conditions of humans are needed for training free-flap harvesting and anastomosis. Animal models for training purposes have been available for years in many surgical fields. We used the female (because they are easy to handle for the procedure) Yorkshire pigs for the head and neck reconstruction by harvesting the deep inferior epigastric artery perforator or the superior epigastric artery perforator flap. The anastomosis site (neck skin defect or tracheal wall defect) was prepared via the dissection of the common carotid artery and the internal jugular vein, in which 3.5&#215; loupe magnification was used for anastomosis as we use on human cases in real life. This procedure demonstrates a new training method using a reliable learning model and provides a detailed anatomy in a live scenario. We focused on the ischemia time, harvesting, vessel anastomosis, and designing the flap to fit the defect site. This model improves tissue handling and with the use of proper instruments can be repeated many times so that the surgeon is fully confident before starting the surgery on humans.

Here we present a protocol for the use of the pig superior epigastric artery perforator flap as a learning module for head and neck microvascular reconstruction.

Porcino come un modulo di formazione per ricostruzione microvascolare del collo e testa

Live models that resemble surgical conditions of humans are needed for training free-flap harvesting and anastomosis. Animal models for training purposes ...

Learning Modern Laryngeal Surgery in a Dissection Laboratory

Surgery for laryngeal malignancies requires millimetric accuracy from the different endoscopic and open techniques available. Practice of this surgery is almost completely reserved to a few referral centers that deal with a large proportion of this pathology. Practice on human specimens is not always possible for ethical, economic, or availability reasons. The aim of this study is to provide a reproducible method for the organization of a laryngeal laboratory on ex vivo animal models where it is possible to approach, learn, and refine laryngeal techniques. Porcine and ovine larynges are ideal, affordable, models to simulate laryngeal surgery given their similarity to the human larynx in their anatomical layout and tissue composition. Herein, the surgical steps of transoral laser surgery, open partial horizontal laryngectomy, and total laryngectomy are reported. The merging of endoscopic and exoscopic views guarantees an inside-out perspective, which is vital for the comprehension of the complex laryngeal anatomy. The method was successfully adopted during three sessions of a dissection course &#34;Lary-Gym&#34;. Further perspectives on robotic surgical training are described.

The purpose of this paper is to illustrate how to organize a reproducible laboratory for laryngeal surgery on affordable and closely similar animal laryngeal models in order to improve anatomical and surgical knowledge and skills.

Imparare la chirurgia larigole moderna in un laboratorio di dissezione

Surgery for laryngeal malignancies requires millimetric accuracy from the different endoscopic and open techniques available. Practice of this surgery is ...

A Revised Method for Inducing Secondary Lymphedema in the Hindlimb of Mice

Animal models are of paramount importance in the research of lymphedema in order to understand the pathophysiology of the disease but also to explore potential treatment options. This mouse model allows researchers to induce significant lymphedema lasting at least 8 weeks. Lymphedema is induced using a combination of fractioned radiotherapy and surgical ablation of lymphatics. This model requires that the mice get a dose of 10 Gray (Gy) radiation before and after surgery. The surgical part of the model involves ligation of three lymph vessels and extraction of two lymph nodes from the mouse hindlimb. Having access to microsurgical tools and a microscope is essential, due to the small anatomical structures of mice. The advantage of this model is that it results in statistically significant lymphedema, which provides a good basis for evaluating different treatment options. It is also a great and easily available option for microsurgical training. The limitation of this model is that the procedure can be time consuming, especially if not practiced in advance. The model results in objectively quantifiable lymphedema in mice, without causing severe morbidity and has been tested in three separate projects.

This animal model enables researchers to induce statistically significant secondary lymphedema in the hindlimb of mice, lasting at least 8 weeks. The model can be used to study the pathophysiology of lymphedema and to investigate novel treatment options.

Un metodo rivisto per indurre il linfomal secondario nell'Hindlimb dei topi

Animal models are of paramount importance in the research of lymphedema in order to understand the pathophysiology of the disease but also to explore ...

Modeling Stroke in Mice: Focal Cortical Lesions by Photothrombosis

Stroke is a leading cause of death and acquired adult disability in developed countries. Despite extensive investigation for novel therapeutic strategies, there remain limited therapeutic options for stroke patients. Therefore, more research is needed for pathophysiological pathways such as post-stroke inflammation, angiogenesis, neuronal plasticity, and regeneration. Given the inability of in vitro models to reproduce the complexity of the brain, experimental stroke models are essential for the analysis and subsequent evaluation of novel drug targets for these mechanisms. In addition, detailed standardized models for all procedures are urgently needed to overcome the so-called replication crisis. As an effort within the ImmunoStroke research consortium, a standardized photothrombotic mouse model using an intraperitoneal injection of Rose Bengal and the illumination of the intact skull with a 561 nm laser is described. This model allows the performance of stroke in mice with allocation to any cortical region of the brain without invasive surgery; thus, enabling the study of stroke in various areas of the brain. In this video, the surgical methods of stroke induction in the photothrombotic model along with&#160;histological analysis are demonstrated.

Described here is the photothrombotic stroke model, where a stroke is produced through the intact skull by inducing permanent microvascular occlusion using laser illumination after administration of a photosensitive dye.

Modellazione dell'ictus nei topi: lesioni corticali focali da fototrombosi

Stroke is a leading cause of death and acquired adult disability in developed countries. Despite extensive investigation for novel therapeutic strategies, ...

Neuroscienze

Generating Murine Model of Myocardial Infarction: A Surgical Procedure for Permanent Ligation of Left Anterior Descending Coronary Artery in Mouse Model

Source: Lugrin, J. et al. <a href="https://www-jove-com.bdigitaluss.remotexs.co/video/59591" target="_blank">Murine Myocardial Infarction Model using Permanent Ligation of Left Anterior Descending Coronary Artery</a>. J. Vis. Exp. (2019)
In this video, we describe a surgical procedure to generate a mouse model of myocardial infarction by permanently ligating the left anterior descending coronary artery.

Generazione di un modello murino di infarto del miocardio: una procedura chirurgica per la legatura permanente dell'arteria coronaria discendente anteriore sinistra in un modello murino

Source: Lugrin, J. et al. Murine Myocardial Infarction Model using Permanent Ligation of Left Anterior Descending Coronary Artery. J. Vis. Exp. (2019)
In ...

JoVE Encyclopedia of Experiments

Biologia

Rodent Models

Adult

Fluorescence-guided Tumor Resection and MSC-seeded Scaffold Implantation in Murine Model: A Surgical Procedure to Deliver Therapeutic Mesenchymal Stem Cells into Brain Tumor Resection Cavity in Mouse Model

Source: Sheets, K. T. et al. <a href="https://www-jove-com.bdigitaluss.remotexs.co/video/57452" target="_blank">Image-Guided Resection of Glioblastoma and Intracranial Implantation of Therapeutic Stem Cell-seeded Scaffolds.</a>&#160;J. Vis. Exp.&#160;(2018)
This video presents the surgical procedure for image-guided tumor resection and implantation of a bio-scaffold seeded with genetically engineered mesenchymal stem cells or MSCs expressing tumor necrosis factor-related apoptosis-inducing ligand or TRAIL. These therapeutic MSCs have the potential in treatment of brain tumors.

Resezione tumorale guidata da fluorescenza e impianto di scaffold seminato con MSC in modello murino: una procedura chirurgica per fornire cellule staminali mesenchimali terapeutiche nella cavità di resezione del tumore cerebrale in un modello murino

Source: Sheets, K. T. et al. Image-Guided Resection of Glioblastoma and Intracranial Implantation of Therapeutic Stem Cell-seeded Scaffolds.&#160;J. Vis. ...

Ricerca sul cancro

Cancers of the Nervous System

In vivo studies

Filament Insertion-based Transient Stroke Model: A Surgical Procedure to Perform Middle Cerebral Artery Occlusion in Murine Brain

Source: Llovera, G., et al. <a href="https://www-jove-com.bdigitaluss.remotexs.co/v/62573">Modeling Stroke in Mice: Transient Middle Cerebral Artery Occlusion via the External Carotid Artery.</a> J. Vis. Exp (2021).
This video describes the procedure to develop transient stroke in a murine model using silicon-coated filament-based occlusion of the middle cerebral artery.&#160;The developed model mimics human stroke, thus helping to investigate novel therapeutic strategies for treating stroke.

Modello di ictus transitorio basato sull'inserimento di filamenti: una procedura chirurgica per eseguire l'occlusione dell'arteria cerebrale media nel cervello murino

Source: Llovera, G., et al. Modeling Stroke in Mice: Transient Middle Cerebral Artery Occlusion via the External Carotid Artery. J. Vis. Exp (2021).
This ...