Un ringraziamento speciale a Chris Kassotis e Audrey Bailey per la revisione critica di questo manoscritto e il Dr. Scott Korte, Giuseppe Beeman, Alison Curfman, Paul Kimball, Bridget Neibreggue, Jacob Redel, Amy Schroder, Maija Steinberg, e Stacey Winkeler per la loro assistenza per l&#39;ottimizzazione di questo modello nel nostro laboratorio. Il finanziamento è stato fornito dalla formazione di Grant Clinica Biodetectives (NIH T90) (Kep), Università del Missouri Scienze della Vita Undergraduate Research Program Opportunità, MU Research Council, MU Assegni di ricerca Board e NIH R21HD056441 (SCN).

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E., Edwards, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+superovulation+and+pregnancy+in+mature+mice+by+gonadotrophins.">Induction of superovulation and pregnancy in mature mice by gonadotrophins.</a> J. Endocrinol. 15, 374-384 (1957).</li><li>Wilson, E. D., Zarrow, M. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+superovulation+in+the+immature+mouse+and+rat.">Comparison of superovulation in the immature mouse and rat.</a> J. Reprod. Fertil. 3, 148-158 (1962).</li><li>Lee, B., Du, H., Taylor, H. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+murine+endometriosis+induces+DNA+methylation+and+altered+gene+expression+in+eutopic+endometrium.">Experimental murine endometriosis induces DNA methylation and altered gene expression in eutopic endometrium.</a> Biol. Reprod. 80, 79-85 (2009).</li><li>Somigliana, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endometrial+ability+to+implant+in+ectopic+sites+can+be+prevented+by+interleukin-12+in+a+murine+model+of+endometriosis.">Endometrial ability to implant in ectopic sites can be prevented by interleukin-12 in a murine model of endometriosis.</a> Hum. 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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endometriotic+lesions+synthesize+and+secrete+a+haptoglobin-like+protein.">Endometriotic lesions synthesize and secrete a haptoglobin-like protein.</a> Biol. Reprod. 58, 988-994 (1998).</li><li>Yavuz, E., Oktem, M., Esinler, I., Toru, S. A., Zeyneloglu, H. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genistein+causes+regression+of+endometriotic+implants+in+the+rat+model.">Genistein causes regression of endometriotic implants in the rat model.</a> Fertil. Steril. 88, 1129-1134 (2007).</li><li>Dmitrieva, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endocannabinoid+involvement+in+endometriosis.">Endocannabinoid involvement in endometriosis.</a> Pain. 151, 703-710 (2010).</li><li>Efstathiou, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nonsteroidal+antiinflammatory+drugs+differentially+suppress+endometriosis+in+a+murine+model.">Nonsteroidal antiinflammatory drugs differentially suppress endometriosis in a murine model.</a> Fertil. Steril. 83, 171-181 (2005).</li><li>Becker, C. 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Nessun conflitto di interessi dichiarati.

Ci sono diversi parametri critici che dovrebbero essere notato durante l&#39;esecuzione l&#39;induzione chirurgica di endometriosi nei topi. In primo luogo, l&#39;endometriosi è una malattia estrogeno-dipendenti e come tale questo tipo di chirurgia deve essere eseguita nell&#39;animale intatto o in alternativa negli animali ovariectomizzati integrato con estrogeni 20. In secondo luogo, la sutura biopsie endometriali alla cascata arteriosa deve essere eseguita con estrema cura. Abbiamo scoperto che usando sol...

<table border="1"><tbody><tr><td> Nome del reagente / apparecchiatura </td><td> Azienda </td><td> Numero di catalogo </td></tr><tr><td> Matita di cera </td><td> Pescatore </td><td> NC9954135 </td></tr><tr><td> Vetrini </td><td> Pescatore </td><td> 12-550-433 </td></tr><tr><td> Contagocce </td><td> Pescatore </td><td> S79383 </td></tr><tr><td> Microscopio ottico standard per la valutazione strisci citologia vaginale </td><td></td><td></td></tr><tr><td> Buprenorfina HCL c3 (CARJET) 10X1ml </td><td> Butler animali Alimentazione Salute </td><td> 022891 </td></tr><tr><td> Fosfato sterile salina tamponata (PBS) </td><td> Gibco </td><td> 14040-117 </td></tr><tr><td> 10.000 U / ml penicillina, 10.000 mg / ml di streptomicina in 0,85% NaCl </td><td> Hyclone </td><td> SV30010 </td></tr><tr><td> Isoflurano </td><td> AbbottAnimal Health </td><td> 05260-05 </td></tr><tr><td> Isoflurano non rirespirazione anestetico sistema </td><td></td><td></td></tr><tr><td> Ricircolo di acqua calda di riscaldamento pad </td><td></td><td></td></tr><tr><td> 30 ml siringa guaina </td><td> Pescatore </td><td> 14-823-16G </td></tr><tr><td> Polvere di guanti sterili libero </td><td> Fisherbrand </td><td> 19020558 </td></tr><tr><td> Unguento oftalmico </td><td> Farmaceutica principali </td><td> 10033691 </td></tr><tr><td> Piccolo tagliaunghie elettrici </td><td> Wahl </td><td> 9861-600 </td></tr><tr><td> Clorexidina macchia </td><td> Pescatore </td><td> NC9863042 </td></tr><tr><td> 70% di etanolo </td><td></td><td></td></tr><tr><td> Polylined campo sterile </td><td> Busse Ospedale Disposables </td><td> 696 </td></tr><tr><td> Taglia 3 bisturi </td><td> Pescatore </td><td> 22-079-657 </td></tr><tr><td> Numero 10 lame bisturi </td><td> Pescatore </td><td> 22-079-681 </td></tr><tr><td> Piccole forbici chirurgiche </td><td> Roboz </td><td> RS-5850 </td></tr><tr><td> Piccoli semi-curve seghettate forcipe </td><td> Roboz </td><td> RS-5135 </td></tr><tr><td> 5-0 di seta nera intrecciata sutura </td><td> Ethicon </td><td> K870H </td></tr><tr><td> Sterilizzato vetro pirex Petri </td><td> Corning </td><td> 70160-101 </td></tr><tr><td> 2 millimetri biopsia </td><td> Miltex </td><td> 33-31 </td></tr><tr><td> Garza sterile </td><td> Kendall </td><td> 1806 </td></tr><tr><td> 6-0 nero ethilon sutura monofilo di nylon </td><td> Ethicon </td><td> 697G </td></tr><tr><td> Driver ago (opzionale) </td><td> Mondo Strumenti di precisione </td><td> 500023 </td></tr><tr><td> 5-0 undyed intrecciato rivestito vicryl sutura </td><td> Ethicon </td><td> J490G </td></tr><tr><td> 9 millimetri Autoclip ferita clip </td><td> Becton Dickinson </td><td> 427631 </td></tr><tr><td> Autoclip applicatore e di rimozione </td><td> Becton Dickinson </td><td> 427630 </td></tr><tr><td> 23G ago </td><td> Becton Dickinson </td><td> 305193 </td></tr><tr><td> 1cc siringa </td><td> Becton Dickson </td><td> 301025 </td></tr><tr><td> 5X vetro stare ingrandimento (opzionale) </td><td> Pescatore </td><td> 14-648-23 </td></tr><tr><td> 10% di formalina tamponata </td><td> Pescatore </td><td> SF100-4 </td></tr><tr><td> Pinze </td><td> Roboz </td><td> RS-6466 </td></tr><tr><td> Elaborazione / embedding cassette </td><td> Pescatore </td><td> 15-197-700A </td></tr><tr><td> Biopsia cuscinetti in schiuma </td><td> Pescatore </td><td> 22-038-222 </td></tr><tr><td> RNAqueous RNA Kit di isolamento </td><td>Ambion </td><td> AM1912 </td></tr><tr><td> Azoto liquido </td><td></td><td></td></tr><tr><td> Snap microcentrifuga coppola tubo orizzontale </td><td> Pescatore </td><td> 02-681-240 </td></tr><tr><td> Ketamina (opzionale) </td><td> Simga </td><td> K4138 </td></tr><tr><td> Domitor (medetomidina cloridrato) (opzionale) </td><td> Tocris </td><td> 2023 </td></tr><tr><td> Antisedan (atipamezolo) (opzionale) </td><td> Sigma </td><td> A9611 </td></tr></tbody></table>

mouse model of surgically-induced endometriosis by auto-transplantation of uterine tissue

L&#39;endometriosi è una malattia cronica, dolorosa malattia la cui eziologia rimane sconosciuta. Inoltre, il trattamento di endometriosi può richiedere la rimozione laparoscopica delle lesioni, e / o croniche gestione farmaceutica del dolore e dei sintomi infertilità. Il costo associato con endometriosi è stato stimato a 22 miliardi di dollari all&#39;anno negli Stati Uniti 1. Per ulteriormente la nostra comprensione dei meccanismi alla base di questa malattia enigmatica, i modelli animali sono stati impiegati. Primati sviluppano spontaneamente endometriosi e quindi modelli di primati più simili alla malattia nelle donne. Modelli di roditori, tuttavia, sono più convenienti e facilmente reperibili 2. Il modello che abbiamo descritto qui comporta un trasferimento autologo di tessuto uterino al mesentere intestinale (Figura 1) ed è stato sviluppato nel ratto 3 e poi trasferito al mouse 4. L&#39;obiettivo del modello roditore autologo di endometriosi chirurgicamente indotta è quella di imitarela malattia nelle donne. Noi e altri hanno già dimostrato che l&#39;alterato pattern di espressione genica osservata nelle lesioni endometriosiche da topi o ratti rispecchia quella osservata nelle donne con la malattia 5,6. Uno dei vantaggi del realizzare l&#39;intervento nel topo è che l&#39;abbondanza di ceppi di topi transgenici disponibile può aiutare i ricercatori a determinare il ruolo di specifici componenti importanti nella creazione e la crescita di endometriosi. Un modello alternativo in cui vengono introdotti asportati frammenti umani endometriale al peritoneo di topi immunocompromessi è anche ampiamente utilizzato, ma è limitata dalla mancanza di un normale sistema immunitario che si ritiene essere importante per l&#39;endometriosi 2,7. È importante sottolineare che il modello murino di endometriosi indotta chirurgicamente è un modello versatile che è stato utilizzato per studiare come il sistema immunitario 8, 9,10 ormoni e fattori ambientali influenzano 11,12 endometriosi così come gli effetti dell&#39;endometriosi sulla Fertility 13 e il dolore 14.

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Mouse Model of Surgically-induced Endometriosis by Auto-transplantation of Uterine Tissue

Una descrizione della induzione chirurgica di endometriosi in topi e ratti con auto-trapianto di tessuto uterino alla cascata delle arterie del mesentere intestinale.

Modello murino di endometriosi chirurgicamente indotto da Auto-trapianto di tessuto uterino

Endometriosis is a chronic, painful disease whose etiology remains unknown. Furthermore, treatment of endometriosis can require laparoscopic removal of ...

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28.3K Views.  University of Missouri. Una descrizione della induzione chirurgica di endometriosi in topi e ratti con auto-trapianto di tessuto uterino alla cascata delle arterie del mesentere intestinale.

Video: Mouse Model of Surgically-induced Endometriosis by Auto-transplantation of Uterine Tissue

Mouse Model of Intraluminal MCAO: Cerebral Infarct Evaluation by Cresyl Violet Staining

Stroke is the third cause of mortality and the leading cause of disability in the World. Ischemic stroke accounts for approximately 80% of all strokes. However, the thrombolytic tissue plasminogen activator (tPA) is the only treatment of acute ischemic stroke that exists. This led researchers to develop several ischemic stroke models in a variety of species. Two major types of rodent models have been developed: models of global cerebral ischemia or focal cerebral ischemia. To mimic ischemic stroke in patients, in whom approximately 80% thrombotic or embolic strokes occur in the territory of the middle cerebral artery (MCA), the intraluminal middle cerebral artery occlusion (MCAO) model is quite relevant for stroke studies. This model was first developed in rats by Koizumi et al. in 1986 1. Because of the ease of genetic manipulation in mice, these models have also been developed in this species 2-3.
	Herein, we present the transient MCA occlusion procedure in C57/Bl6 mice. Previous studies have reported that physical properties of the occluder such as tip diameter, length, shape, and flexibility are critical for the reproducibility of the infarct volume 4. Herein, a commercial silicon coated monofilaments (Doccol Corporation) have been used. Another great advantage is that this monofilament reduces the risk to induce subarachnoid hemorrhages. Using the Zeiss stereo-microscope Stemi 2000, the silicon coated monofilament was introduced into the internal carotid artery (ICA) via a cut in the external carotid artery (ECA) until the monofilament occludes the base of the MCA. Blood flow was restored 1 hour later by removal of the monofilament to mimic the restoration of blood flow after lysis of a thromboembolic clot in humans. The extent of cerebral infarct may be evaluated first by a neurologic score and by the measurement of the infarct volume. Ischemic mice were thus analyzed for their neurologic score at different post-reperfusion times. To evaluate the infarct volume, staining with 2,3,5-triphenyltetrazolium chloride (TTC) was usually performed. Herein, we used cresyl violet staining since it offers the opportunity to test many critical markers by immunohistochemistry. In this video, we report the MCAO procedure; neurological scores and the evaluation of the infarct volume by cresyl violet staining.

The intraluminal middle cerebral occlusion model in mice is herein presented. The extent of cerebral infarct is evaluated by a neurologic score and cresyl violet staining, an alternative staining to TTC, offering the great advantage to test in parallel many interest markers.

Modello murino di MCAO intraluminale: valutazione infarto cerebrale da colorazione viola cresile

Stroke is the third  cause of mortality and the leading cause of disability in the World. Ischemic  stroke accounts for approximately 80% of all strokes. ...

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Assessment of Right Ventricular Structure and Function in Mouse Model of Pulmonary Artery Constriction by Transthoracic Echocardiography

Emerging clinical data support the notion that RV dysfunction is critical to the pathogenesis of cardiovascular disease and heart failure1-3. Moreover, the RV is significantly affected in pulmonary diseases such as pulmonary artery hypertension (PAH). In addition, the RV is remarkably sensitive to cardiac pathologies, including left ventricular (LV) dysfunction, valvular disease or RV infarction4. To understand the role of RV in the pathogenesis of cardiac diseases, a reliable and noninvasive method to access the RV structurally and functionally is essential.
A noninvasive trans-thoracic echocardiography (TTE) based methodology was established and validated for monitoring dynamic changes in RV structure and function in adult mice. To impose RV stress, we employed a surgical model of pulmonary artery constriction (PAC) and measured the RV response over a 7-day period using a high-frequency ultrasound microimaging system. Sham operated mice were used as controls. Images were acquired in lightly anesthetized mice at baseline (before surgery), day 0 (immediately post-surgery), day 3, and day 7 (post-surgery). Data was analyzed offline using software.
Several acoustic windows (B, M, and Color Doppler modes), which can be consistently obtained in mice, allowed for reliable and reproducible measurement of RV structure (including RV wall thickness, end-diastolic&#160;and end-systolic dimensions), and function (fractional area change, fractional shortening, PA peak velocity, and peak pressure gradient) in normal mice and following PAC.
Using this method, the pressure-gradient resulting from PAC was accurately measured in real-time using Color Doppler mode and was comparable to direct pressure measurements performed with a Millar high-fidelity microtip catheter. Taken together, these data demonstrate that RV measurements obtained from various complimentary views using echocardiography are reliable, reproducible and can provide insights regarding RV structure and function. This method will enable a better understanding of the role of RV cardiac dysfunction.

Right ventricle (RV) dysfunction is critical to the pathogenesis of cardiovascular disease, yet limited methodologies are available for its evaluation. Recent advances in ultrasound imaging provide a noninvasive and accurate option for longitudinal RV study. Herein, we detail a step-by-step echocardiographic method using a murine model of RV pressure overload.

Valutazione del ventricolo destro Struttura e funzione in modello murino di polmonare Arteria Costrizione da ecocardiografia transtoracica

Emerging clinical data support the notion that RV dysfunction is critical to the pathogenesis of cardiovascular disease and heart failure1-3. Moreover, ...

Implantation of Inferior Vena Cava Interposition Graft in Mouse Model

Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are often used for reconstructive surgery to treat congenital cardiac anomalies. The long-term clinical results showed excellent patency rates, however, with significant incidence of stenosis. To investigate the cellular and molecular mechanisms of vascular neotissue formation and prevent stenosis development in tissue engineered vascular grafts (TEVGs), we developed a mouse model of the graft with approximately 1 mm internal diameter. First, the TEVGs were assembled from biodegradable tubular scaffolds fabricated from a polyglycolic acid nonwoven felt mesh coated with &#949;-caprolactone and L-lactide copolymer. The scaffolds were then placed in a lyophilizer, vacuumed for 24 hr, and stored in a desiccator until cell seeding. Second, bone marrow was collected from donor mice and mononuclear cells were isolated by density gradient centrifugation. Third, approximately one million cells were seeded on a scaffold and incubated O/N. Finally, the seeded scaffolds were then implanted as infrarenal vena cava interposition grafts in C57BL/6 mice. The implanted grafts demonstrated excellent patency (&#62;90%) without evidence of thromboembolic complications or aneurysmal formation. This murine model will aid us in understanding and quantifying the cellular and molecular mechanisms of neotissue formation in the TEVG.

To improve our knowledge of cellular and molecular neotissue formation, a murine model of the TEVG was recently developed. The grafts were implanted as infrarenal vena cava interposition grafts in C57BL/6 mice. This model achieves similar results to those achieved in our clinical investigation, but over a far shortened time-course.

L&#39;impianto di Vena cava inferiore Interposizione Graft in modello murino

Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are often used for reconstructive surgery to treat congenital cardiac anomalies. ...

Transplantation of Pulmonary Valve Using a Mouse Model of Heterotopic Heart Transplantation

Tissue engineered heart valves, especially decellularized valves, are starting to gain momentum in clinical use of reconstructive surgery with mixed results. However, the cellular and molecular mechanisms of the neotissue development, valve thickening, and stenosis development are not researched extensively. To answer the above questions, we developed a murine heterotopic heart valve transplantation model. A heart valve was harvested from a valve donor mouse and transplanted to a heart donor mouse. The heart with a new valve was transplanted heterotopically to a recipient mouse. The transplanted heart showed its own heartbeat, independent of the recipient&#8217;s heartbeat. The blood flow was quantified using a high frequency ultrasound system with a pulsed wave Doppler. The flow through the implanted pulmonary valve showed forward flow with minimal regurgitation and the peak flow was close to 100 mm/sec. This murine model of heart valve transplantation is highly versatile, so it can be modified and adapted to provide different hemodynamic environments and/or can be used with various transgenic mice to study neotissue development in a tissue engineered heart valve.

In order to understand the cellular and molecular mechanisms underlying neotissue formation and stenosis development in tissue engineered heart valves, a murine model of heterotopic heart valve transplantation was developed. A pulmonary heart valve was transplanted to recipient using the heterotopic heart transplantation technique.

Il trapianto di valvola polmonare Utilizzando un modello murino di trapianto cardiaco eterotopico

Tissue engineered heart valves, especially decellularized valves, are starting to gain momentum in clinical use of reconstructive surgery with mixed ...

Rose Bengal Photothrombosis by Confocal Optical Imaging In Vivo: A Model of Single Vessel Stroke

In vivo imaging techniques have increased in utilization due to recent advances in imaging dyes and optical technologies, allowing for the ability to image cellular events in an intact animal. Additionally, the ability to induce physiological disease states such as stroke in vivo increases its utility. The technique described herein allows for physiological assessment of cellular responses within the CNS following a stroke and can be adapted for other pathological conditions being studied. The technique presented uses laser excitation of the photosensitive dye Rose Bengal in vivo to induce a focal ischemic event in a single blood vessel. 
The video protocol demonstrates the preparation of a thin-skulled cranial window over the somatosensory cortex in a mouse for the induction of a Rose Bengal photothrombotic event keeping injury to the underlying dura matter and brain at a minimum. Surgical preparation is initially performed under a dissecting microscope with a custom-made surgical/imaging platform, which is then transferred to a confocal microscope equipped with an inverted objective adaptor. Representative images acquired utilizing this protocol are presented as well as time-lapse sequences of stroke induction. This technique is powerful in that the same area can be imaged repeatedly on subsequent days facilitating longitudinal in vivo studies of pathological processes following stroke.

Rose Bengal Photothrombosis by Confocal Optical Imaging In Vivo:  A Model of Single Vessel Stroke

Here, we describe a semi-invasive optical microscopy approach for the induction of a Rose Bengal photothrombotic clot in the somatosensory cortex of a mouse in vivo. The technical aspects of the imaging procedure are described from induction of a photothrombotic event to application and data collection.

Rosa Bengala Photothrombosis da confocale Optical Imaging<em&gt; In Vivo</em&gt;: Un modello di singolo vaso Stroke

In vivo imaging techniques have increased in utilization due to recent advances in imaging dyes and optical technologies, allowing for the ability to ...

Creation of a Rodent Model of Abdominal Aortic Aneurysm by Blocking Adventitial Vasa Vasorum Perfusion

The adventitial vasa vasorum (VV) provides oxygen and nourishment to the aortic wall. Hypoxia in the aortic wall can cause enlarged abdominal aortic aneurysms (AAAs). This article introduces and describes a standard protocol used to induce AAAs through adventitial VV hypoperfusion created with a combination of polyurethane catheter insertion into the aortic lumen and suture ligation of the infrarenal abdominal aorta.
The protocol involves the use of male rats weighing 300-400 g, which are provided food and water ad libitum. After laparotomy with a ventral midline abdominal incision, exfoliation of the aorta is performed, which blocks blood flow from the perivascular tissue. Aortotomy involving a small incision adjacent to the renal artery branches is performed, and a polyurethane catheter is inserted using an 18-gauge indwelling needle. After repairing the incision, tight ligation of the aorta over the catheter blocks VV blood flow from the proximal direction through the aortic wall without disturbing the aortic blood flow. This technique can induce an AAA with progressive aortic dilatation.
The greatest benefit of this model is that VV hypoperfusion causes tissue hypoxia and the development of an infrarenal AAA, which has morphological and pathological characteristics similar to those of a human AAA.

Polyurethane catheter insertion into the aortic lumen and suture ligation of the aorta induce chronic hypoxia due to hypoperfusion of the adventitial vasa vasorum. This article describes a novel animal model of abdominal aortic aneurysm (AAA) with characteristics similar to those of AAA in humans.

Creazione di un modello del roditore del Aneurysm aortico addominale bloccando Adventitial Vasa Vasorum perfusione

The adventitial vasa vasorum (VV) provides oxygen and nourishment to the aortic wall. Hypoxia in the aortic wall can cause enlarged abdominal aortic ...

A Mouse Model of Orthopedic Surgery to Study Postoperative Cognitive Dysfunction and Tissue Regeneration

Surgery is commonly used to improve and maintain quality of life. Unfortunately, in vulnerable patients such as the elderly, complications may occur and significantly diminish the outcome. Indeed, after routine orthopedic surgery to repair a fracture, as many as 50% of elderly patients suffer from neurologic complications like delirium. Also, the capacity to heal and regenerate tissue after surgery decreases with age, and can impact the quality of fracture repair and even osseous integration of implants. Thus, a better understanding of mechanisms that drive these age-dependent changes could provide strategic targets to minimize risk for such complications and optimize outcomes. Here, we introduce a clinically relevant mouse model of tibial fracture. The postoperative changes in these mice mimic some of the cognitive impairments commonly observed after routine orthopedic surgery in humans. Briefly, an incision is performed in the right hind limb under strictly aseptic conditions. Muscles are disassociated, and a 0.38-mm stainless steel pin is inserted into the upper crest of the tibia, inside the intramedullary canal. Osteotomy is then performed, and the wound is stapled. We have used this model to investigate the effects of surgical trauma on postoperative neuroinflammation and behavioral changes. By applying this fracture model in combination with parabiosis, a surgical model in which 2 mice are anastomosed, we have studied cells and secreted factors that systemically rejuvenate organ function and tissue regeneration after injury. By following our step-by-step protocol, these models can be reproduced with high fidelity, and can be adapted to interrogate many biologic pathways that are altered by surgical trauma.

This protocol describes a mouse model of orthopedic surgery that has been used to study mechanisms of postoperative neuroinflammation and behavioral changes, and when combined with parabiosis, to study tissue regeneration during aging.

Un modello murino di chirurgia ortopedica per studiare la disfunzione cognitiva postoperatoria e rigenerazione tissutale

Surgery is commonly used to improve and maintain quality of life. Unfortunately, in vulnerable patients such as the elderly, complications may occur and ...

Improvement of a Closed Chest Porcine Myocardial Infarction Model by Standardization of Tissue and Blood Sampling Procedures

Myocardial ischemia reperfusion (I/R) injury contributes to almost half of the necrotic area after myocardial infarction. To date there is no approved drug to prevent or reduce myocardial I/R injury. The study and understanding of the pathophysiological mechanisms of myocardial I/R injury is essential to develop successful treatments. Large animal experiments are an important step in translational methods. The porcine model of acute myocardial infarction has been established and described by ourselves and others. We aimed to further improve the value of the model by focusing in detail on the sampling techniques for use in future experiments. Furthermore, we emphasize small but important steps that can affect the quality of the final results. To mimic the clinical situation of myocardial I/R injury, a percutaneous coronary intervention (PCI) catheter was inserted into the left anterior descending coronary artery (LAD) of an anesthetized pig. &#176;&#176;&#176;This model mimics acute myocardial infarction and PCI treatment in humans with the possibility of accurately determining the area at risk as well as the necrotic- and viable ischemic tissue. Here the model was used to investigate the effect of a bicyclic peptide inhibitor of FXIIa. The model can also be modified to allow longer reperfusion times to study later effects of myocardial infarction.

Here we demonstrate a protocol to standardize sampling procedures of an established porcine model of acute myocardial infarction in order to increase its translational value in the understanding of the pathophysiology of myocardial ischemia/reperfusion injury and to test novel drug candidates.

Miglioramento di un modello di infarto miocardico porcina cassa chiusa tramite la normalizzazione del tessuto e le procedure di campionamento di sangue

Myocardial ischemia reperfusion (I/R) injury contributes to almost half of the necrotic area after myocardial infarction. To date there is no approved ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

Misurazione di contrattilità isometrica dell'arteria mesenterica Mouse con filo Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

A Mouse Model of Incompletely Resected Soft Tissue Sarcoma for Testing (Neo)adjuvant Therapies

Surgery is often the first treatment for many solid tumors. However, local relapses frequently occur following primary tumor resection, despite adjuvant or neo-adjuvant therapies. This occurs when surgical margins are insufficiently tumor-free, resulting in residual cancer cells. From a biological and immunological perspective, surgery is not a null event; the wound healing environment is known to induce both pro- and anti-tumorigenic pathways. As a consequence, preclinical models for drug development aimed at preventing local relapse should incorporate surgical resection when testing new (neo)adjuvant therapies, to model the clinical settings in patients treated with surgery.
Here, we describe a mouse model of incomplete surgical resection of WEHI 164 soft tissue sarcoma that allows testing of (neo)adjuvant therapies in the setting of a wound healing response. In this model, 50% or 75% of the tumor is removed, leaving behind some cancer tissue in situ to model gross residual disease after surgery in the clinical setting. This model allows testing therapies in the context of surgery while also considering the wound healing response, which may affect the efficacy of (neo)adjuvant treatments. The incomplete surgical resection results in reproducible regrowth of the tumor in all mice in the absence of adjuvant therapy. Adjuvant treatment with checkpoint blockade results in reduced tumor regrowth. This model is thus appropriate for testing therapies in the context of debulking surgery and its associated wound healing response and can be extended to other types of solid cancer.

A Mouse Model of Incompletely Resected Soft Tissue Sarcoma for Testing Neoadjuvant Therapies

In this protocol, we describe a mouse model of incomplete surgical resection of soft tissue sarcoma for testing (neo)adjuvant therapies.

Un modello di topo di Sarcoma di tessuto molle incompleto resected per test (Neo)adiuvante Terapie

Surgery is often the first treatment for many solid tumors. However, local relapses frequently occur following primary tumor resection, despite adjuvant ...