Gli autori sono grati al Dott. Ana Cotrim per la consulenza tecnica e al Dott. Monzur Murshed per le attrezzature. Questo lavoro è stato in parte finanziato dal Canadian Institutes per la ricerca medica.

1. Descrizione procedurali <ol><li> Iniettare per via sottocutanea 1 ml / g di peso corporeo di 0,5 mg / ml monoidrato atropina solfato (Cat # 11330, Fluka, USA) in ogni mouse per evitare che le secrezioni salivari di compromettere il movimento delle infusioni. </li><li> Anestetizzare ogni mouse con 1 peso corporeo microlitri / g di un mg / 100 ml di ketamina e 20 mg / ml xylazina (Phoenix scientifico, AR, USA) soluzione iniettata intraperitoneale (IP) </li><li> Posizionare il mouse su una misura piattaforma in plastica in posizione ventrale: incisivi superiori sono bloccati su un filo metallico, e gli incisivi mandibolari sono agganciati su una corda elastica per tenere la bocca aperta. </li><li> Utilizzare una siringa per insulina con una 29-gauge (Cat # 600145, Tyco Healthcare, MA, USA) posto all&#39;interno di un tubo in polietilene 0,58 mm di diametro (Cat # 427410, Becton Dickinson, MD, USA; Fig. 2). Il tubo in polietilene è inserita 5mm rispetto alla punta di una 29-gauge. L&#39;ago viene utilizzato come una guida rigida per spostare il tubo in polietilene nel condotto della Wharton. Il tubo in polietilene è inserita 3-5 mm all&#39;interno del condotto. Da notare che solo il tubo in polietilene entra nel condotto della ghiandola salivare mentre l&#39;ago rimane al di fuori del condotto. Iniettare 50ul del filtrata 0,4% soluzione trypan blu macchia (Cat # 15250, Gibco, USA) nella ghiandola sottomandibolare tramite il dotto di Wharton, che si trova proprio sotto la lingua. L&#39;infusion viene consegnato costantemente con pressione positiva sul pistone siringa; il dito rimane sul pistone anche dopo tutti l&#39;infusione è stata consegnata alla ghiandola, al fine di evitare un riflusso di infusione intra-orale. La soluzione trypan blu è usato nel video come colorante per dimostrare la tecnica. </li><li> opo l&#39;iniezione della soluzione trypan blu, il mouse è stato l&#39;eutanasia prima dissezione. La ghiandola salivare è esposto chirurgicamente al fine di determinare la quantità delle infusioni hanno effettivamente rimaste nella ghiandola. </li></ol> 2. Rappresentante dei risultati: Dopo le infusioni sono state iniettate, il mouse è stato sezionato per mostrare chirurgicamente post-cannulata ghiandole salivari. Come mostrato su Immagine-2, la colorazione scura del condotto della Wharton fornisce la prova che la soluzione trypan blu è stato infuso con successo. <img src="/files/ftp_upload/3074/3074fig1.jpg" alt="Figura 1" /> Figura 1. Incannulamento della ghiandola sottomandibolare del mouse. <img src="/files/ftp_upload/3074/3074fig2.jpg" alt="Figura 2" /> Figura 2. Polietilene tubo è racchiuso attorno alla punta di una 29-gauge. L&#39;ago della siringa è usata come una guida rigida per spostare il tubo in polietilene nel condotto della Wharton. Il tubo in polietilene è inserita (cannulata) 3-5 mm all&#39;interno del condotto. Da notare che solo il tubo in polietilene entra nel condotto della ghiandola salivare mentre l&#39;ago rimane al di fuori del condotto.

<ol>
	<li>Fox, P. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acquired+salivary+dysfunction.+Drugs+and+radiation.">Acquired salivary dysfunction. Drugs and radiation.</a> Ann N Y Acad Sci. 842, 132-137 (1998).</li><li>Bhide, S. A., Miah, A. B., Harrington, K. J., Newbold, K. L., Nutting, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radiation-induced+xerostomia:+pathophysiology,+prevention+and+treatment.">Radiation-induced xerostomia: pathophysiology, prevention and treatment.</a> Clin Oncol (R Coll Radiol). 21, 737-744 (2009).</li><li>Redman, R. S., Ball, W. D., Mezey, E., Key, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dispersed+donor+salivary+gland+cells+are+widely+distributed+in+the+recipient+gland+when+infused+up+the+ductal+tree.">Dispersed donor salivary gland cells are widely distributed in the recipient gland when infused up the ductal tree.</a> Biotech Histochem. 84, 253-260 (2009).</li></ol>

Tutti gli esperimenti sono stati approvati dal Comitato di Animal Care Università ed erano in linea con gli orientamenti del Consiglio canadese di prodotti per animali.

Una difficoltà con questo metodo incannulazione è quello di impedire il riflusso della soluzione iniettata. Anche se la soluzione trypan blu ha temporaneamente migrato nella ghiandola salivare con successo, la secrezione della saliva potrebbe interferire con le infusioni e causare riflusso della soluzione trypan blu fuori della ghiandola. Come precauzione, oltre a utilizzare atropina per inibire la secrezione di saliva, la cannula di polietilene dovrebbero rimanere nel canale della Wharton e il pistone della siringa detenuti per 60 secondi dopo le infusioni sono stati depositati. Questo permetterà la maggior parte della soluzione infusa per rimanere all&#39;interno della ghiandola. Convenzionalmente, l&#39;applicazione sistematica, ovvero l&#39;iniezione per via endovenosa (IV) di farmaci / cellule, è stato usato come un approccio clinico per la rigenerazione delle ghiandole salivari. Infusioni per via endovenosa iniettato viaggerà attraverso il sistema circolatorio, consentendo solo alcuni dei farmaci / cellule di migrare nelle ghiandole bersaglio. Rispetto a questo metodo tradizionale, l&#39;applicazione locale, cioè l&#39;iniezione di droghe / cellule tramite la procedura di incannulazione, può essere percepito come un modo più diretto ed efficace nel far rivivere le funzioni della ghiandola parotide, sottomandibolare e salivare sottolinguale. Quindi, questa tecnica incannulazione può essere usato come un modello affidabile per la medicina rigenerativa delle ghiandole salivari del mouse, e come prospettiva futura, questa procedura può essere un metodo utile per l&#39;applicazione clinica. Sarebbe un grande piacere per questo visivo-articolo per essere utilizzato come un efficace strumento per studenti e ricercatori che sono interessati a ulteriori studi in medicina rigenerativa delle ghiandole salivari.

<table border="1"><tbody><tr><td> Nome del reagente </td><td> Tipo </td><td> Azienda </td><td> Numero di catalogo </td><td> Commenti (opzionale) </td></tr><tr><td> Atropina </td><td> Droga </td><td> Fluka </td><td> 11330 </td><td> Per inibire la secrezione della saliva </td></tr><tr><td> Ketamina </td><td> Droga </td><td> Phoenix scientifico </td><td></td><td> Sedazione </td></tr><tr><td> Xylazina </td><td> Droga </td><td> Phoenix scientifico </td><td></td><td></td></tr><tr><td> Tubi in polietilene (0,58 mm di diametro) </td><td> Tubo </td><td> Intramedic PE-10, Becton Dickinson Diagnostic Systems </td><td> 427410 </td><td> Per inserire più di punta dell&#39;ago per evitare lesioni durante l&#39;inserimento nel tessuto del dotto di Wharton. </td></tr><tr><td> Ultra insulina siringa Comfort (con un ago 29G) </td><td> Siringa </td><td> Tyco Healthcare </td><td> 600145 </td><td></td></tr><tr><td> 4% trypan blu </td><td> Macchia </td><td> Gibco </td><td> 15250 </td><td></td></tr></tbody></table>

cannulation of the mouse submandibular salivary gland via the wharton's duct

Grave ipofunzione della ghiandola salivare si trova frequentemente nei pazienti con sindrome di Sjögren e quelli che ricevono l&#39;irradiazione terapeutica nella loro testa e del collo per il trattamento del cancro. Entrambi i gruppi di sintomi esperienza pazienti come xerostomia (secchezza delle fauci), disfagia (compromissione della masticazione e della deglutizione), gravi carie dentale, alterazione del gusto, oro-faringeo infezioni (candidosi), mucosite, dolore e disagio. Un approccio innovativo della medicina rigenerativa per il trattamento delle ghiandole salivari ipo-funzione è speculato in RS Redman, E Mezey et al 2009:. Cellule staminali possono essere depositati direttamente dal incannulazione nella ghiandola come metodo potente nel far rivivere le funzioni della compromissione organo. Presumibilmente, le cellule staminali migrati stranieri differenziarsi in cellule ghiandolari a funzionare come parte della ghiandola salivare ospite. Inoltre, questa tecnica di incannulazione è un metodo di consegna conveniente ed efficace per l&#39;applicazione clinica di trasferimento genico. Qui si illustrano i passaggi necessari per eseguire la procedura incannulazione sulla ghiandola salivare sottomandibolare del mouse tramite il dotto di Wharton (Fig. 1). C3H topi (Charles River, Montreal, QC, Canada) vengono utilizzati per questo esperimento, che sono stati tenuti in condizioni di pulizia convenzionali alla risorsa McGill University centro animale. Tutti gli esperimenti sono stati approvati dal Comitato di Animal Care Università ed erano in linea con gli orientamenti del Consiglio canadese di prodotti per animali. Per questo esperimento, una soluzione trypan blu è infusa nella ghiandola attraverso l&#39;apertura del canale della Wharton usando una siringa per insulina con una 29-gauge racchiuso all&#39;interno di un tubo in polietilene. Successivamente, il mouse è sezionato per mostrare che le infusioni migrati nella ghiandola con successo.

Watch this Scientific Journal Video about Cannulation of the Mouse Submandibular Salivary Gland via the Wharton's Duct at JoVE.com

Cannulation of the Mouse Submandibular Salivary Gland via the Wharton's Duct

Un protocollo per l&#39;incannulazione della ghiandola salivare sottomandibolare del mouse tramite il dotto di Wharton è descritto. Per questo esperimento, la soluzione trypan blu è usato come un tintore per dimostrare quanto questa tecnica offre efficacemente infusi nella ghiandola mirati, e di suggerire l&#39;affidabilità di questo nuovo approccio clinico come un potenziale farmaco / terapia cellulare per la rigenerazione delle ghiandole salivari.

Incannulazione della ghiandola salivare sottomandibolare del mouse tramite il Duct Wharton

Severe salivary gland hypofunction is frequently found in patients with Sj&ouml;gren's syndrome and those who receiving therapeutic 
irradiation in their ...

cannulation-mouse-submandibular-salivary-gland-via-wharton-s

Nanyang Technological University

Research

JoVE Journal

Medicine

25.9K Views.  McGill University. Un protocollo per l'incannulazione della ghiandola salivare sottomandibolare del mouse tramite il dotto di Wharton è descritto. Per questo esperimento, la soluzione trypan blu è usato come un tintore per dimostrare quanto questa tecnica offre efficacemente infusi nella ghiandola mirati, e di suggerire l'affidabilità di questo nuovo approccio clinico come un potenziale farmaco / terapia cellulare per la rigenerazione delle ghiandole salivari.

Video: Cannulation of the Mouse Submandibular Salivary Gland via the Wharton's Duct

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow (ischemia) 1. Middle cerebral artery (MCA) delivers a great majority of blood to the lateral surface of the cortex 2, is the most common site of human stroke 3, and ischemia within its territory can result in extensive dysfunction or death 1,4,5. Survivors of ischemic stroke often suffer loss or disruption of motor capabilities, sensory deficits, and infarct. In an effort to capture these key characteristics of stroke, and thereby develop effective treatment, a great deal of emphasis is placed upon animal models of ischemia in MCA.
Here we present a method of permanently occluding a cortical surface blood vessel. We will present this method using an example of a relevant vessel occlusion that models the most common type, location, and outcome of human stroke, permanent middle cerebral artery occlusion (pMCAO). In this model, we surgically expose MCA in the adult rat and subsequently occlude via double ligature and transection of the vessel. This pMCAO blocks the proximal cortical branch of MCA, causing ischemia in all of MCA cortical territory, a large portion of the cortex. This method of occlusion can also be used to occlude more distal portions of cortical vessels in order to achieve more focal ischemia targeting a smaller region of cortex. The primary disadvantages of pMCAO are that the surgical procedure is somewhat invasive as a small craniotomy is required to access MCA, though this results in minimal tissue damage. The primary advantages of this model, however, are: the site of occlusion is well defined, the degree of blood flow reduction is consistent, functional and neurological impairment occurs rapidly, infarct size is consistent, and the high rate of survival allows for long-term chronic assessment.

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

We describe a highly reproducible method for the permanent occlusion of a rodent major cerebral blood vessel. This technique can be accomplished with very little peripheral damage, minimal blood loss, a high rate of long-term survival, and consistent infarct volume commensurate with the human clinical population.

Cerebrale permanente occlusione di vasi<em&gt; Via</em&gt; Doppio Legatura e resezione

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow ...

Ricerca

Medicina

Technical Aspects of the Mouse Aortocaval Fistula

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta and inferior vena cava (IVC) are exposed. The proximal infrarenal aorta and the distal aorta are dissected for clamp placement and needle puncture, respectively. Special attention is paid to avoid dissection between the aorta and the IVC. After clamping the aorta, a 25 G needle is used to puncture both walls of the aorta into the IVC. The surrounding connective tissue is used for hemostatic compression. Successful creation of the AVF will show pulsatile arterial blood flow in the IVC. Further confirmation of successful AVF can be achieved by post-operative Doppler ultrasound.

The goal is to produce an arteriovenous fistula that is simple and reproducible. This method does not use sutures or glue adhesive. Therefore the samples can be used with the least amount of foreign materials for analysis.

Aspetti tecnici della fistola aorto-cavale mouse

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

Collecting Saliva and Measuring Salivary Cortisol and Alpha-amylase in Frail Community Residing Older Adults via Family Caregivers

Salivary measures have emerged in bio-behavioral research that are easy-to-collect, minimally invasive, and relatively inexpensive biologic markers of stress. This article we present the steps for collection and analysis of two salivary assays in research with frail, community residing older adults-salivary cortisol and salivary alpha amylase. The field of salivary bioscience is rapidly advancing and the purpose of this presentation is to provide an update on the developments for investigators interested in integrating these measures into research on aging. Strategies are presented for instructing family caregivers in collecting saliva in the home, and for conducting laboratory analyses of salivary analytes that have demonstrated feasibility, high compliance, and yield quality specimens. The protocol for sample collection includes: (1) consistent use of collection materials; (2) standardized methods that promote adherence and minimize subject burden; and (3) procedures for controlling certain confounding agents. We also provide strategies for laboratory analyses include: (1) saliva handling and processing; (2) salivary cortisol and salivary alpha amylase assay procedures; and (3) analytic considerations.

We demonstrate: (1) procedures for collection of salivary samples in cognitive impaired older adults by family caregivers in the home setting, (2) procedures for measuring stress activity via salivary cortisol and alpha amylase, and (3) representative profiles. Protocols that allow researchers to study stress-linked processes advance our understanding of biological sensitivity and susceptibility.

Salivary measures have emerged in bio-behavioral research  that are easy-to-collect, minimally invasive, and relatively inexpensive  biologic markers of ...

Mouse Kidney Transplantation: Models of Allograft Rejection

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely replicates both the technical and pathological processes that occur in human renal transplantation. Although mouse models of allogeneic rejection in organs other than the kidney exist, and are more technically feasible, there is evidence that different organs elicit disparate rejection modes and dynamics, for instance the time course of rejection in cardiac and renal allograft differs significantly in certain strain combinations. This model is an attractive tool for many reasons despite its technical challenges. As inbred mouse strain haplotypes are well characterized it is possible to choose donor and recipient combinations to model acute allograft rejection by transplanting across MHC class I and II loci. Conversely by transplanting between strains with similar haplotypes a chronic process can be elicited were the allograft kidney develops interstitial fibrosis and tubular atrophy. We have modified the surgical technique to reduce operating time and improve ease of surgery, however a learning curve still needs to be overcome in order to faithfully replicate the model. This study will provide key points in the surgical procedure and aid the process of establishing this technique.

Here, we present a protocol to study the immunology of rejection. The surgical model presented reports a short operating time and a concise technique. Depending on the donor-recipient strain combination, the transplanted kidney may develop acute cellular rejection or chronic allograft damage, defined by interstitial fibrosis and tubular atrophy.

Mouse trapianto renale: Modelli di Allograft Rifiuto

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely ...

Surgical Injury to the Mouse Pancreas through Ligation of the Pancreatic Duct as a Model for Endocrine and Exocrine Reprogramming and Proliferation

Expansion of pancreatic beta cells in vivo or ex vivo, or generation of beta cells by differentiation from an embryonic or adult stem cell, can provide new expandable sources of beta cells to alleviate the donor scarcity in human islet transplantation as therapy for diabetes. Although recent advances have been made towards this aim, mechanisms that regulate beta cell expansion and differentiation from a stem/progenitor cell remain to be characterized. Here, we describe a protocol for an injury model in the adult mouse pancreas that can function as a tool to study mechanisms of tissue remodeling and beta cell proliferation and differentiation. Partial duct ligation (PDL) is an experimentally induced injury of the rodent pancreas involving surgical ligation of the main pancreatic duct resulting in an obstruction of drainage of exocrine products out of the tail region of the pancreas. The inflicted damage induces acinar atrophy, immune cell infiltration and severe tissue remodeling. We have previously reported the activation of Neurogenin (Ngn) 3 expressing endogenous progenitor-like cells and an increase in beta cell proliferation after PDL. Therefore, PDL provides a basis to study signals involved in beta cell dynamics and the properties of an endocrine progenitor in adult pancreas. Since, it still remains largely unclear, which factors and pathways contribute to beta cell neogenesis and proliferation in PDL, a standardized protocol for PDL will allow for comparison across laboratories.

This protocol describes an injury model involving the surgical ligation of the pancreatic duct in the adult mouse pancreas, resulting in severe injury that establishes an environment that allows beta cell neogenesis and proliferation. This model can be used as a tool to study mechanisms involved in beta cell formation.

Lesioni chirurgica al pancreas mouse attraverso la legatura del Dotto pancreatico come un modello per Endocrine e Esocrine riprogrammazione e la proliferazione

Expansion of pancreatic beta cells in vivo or ex vivo, or generation of beta cells by differentiation from an embryonic or adult stem cell, can provide ...

Retroductal Submandibular Gland Instillation and Localized Fractionated Irradiation in a Rat Model of Salivary Hypofunction

Normal tissues that lie within the portals of radiation are inadvertently damaged. Salivary glands are often injured during head and neck radiotherapy. Irreparable cell damage results in a chronic loss of salivary function that impairs basic oral activities, and increases the risk of oral infections and dental caries. Salivary hypofunction and its complications gravely impact a patient's comfort. Current symptomatic management of the condition is ineffective, and newer therapies to assuage the condition are needed. 
Salivary glands are exocrine glands, which expel their secretions into the mouth via excretory ducts. Cannulation of these ducts provides direct access to the glands. Retroductal delivery of a contrast agent to major salivary glands is a routine out-patient procedure for diagnostic imaging. Using a similar procedure, localized treatment of the glands is feasible. However, performing this technique in preclinical studies with small animals poses unique challenges. In this study we describe the technique of retroductal administration in rat submandibular glands, a procedure that was refined in Dr. Bruce Baum's laboratory (NIH)1, and lay out a procedure for local gland irradiation.

Salivary gland hypofunction, a major adverse effect of head and neck radiotherapy diminishes a patient's quality of life. The demonstration of efficacy of new therapies in animal models is a prerequisite before clinical transition. This protocol describes retroductal administration and local irradiation of rat submandibular glands.

Retroductal ghiandola sottomandibolare instillazione e localizzati frazionato irradiazione in un modello murino di salivare Ipofunzione

Normal tissues that lie within the portals of radiation are inadvertently damaged. Salivary glands are often injured during head and neck radiotherapy. ...

Pancreatic Duct Infusion: An Effective and Selective Method of Drug and Viral Delivery

The pancreas is a bifunctional organ with both endocrine and exocrine components. A number of pathologies can afflict the pancreas, including diabetes, pancreatitis, and pancreatic cancer. All three of these diseases mark active areas of study, not only to develop immediate therapy, but also to better understand their pathophysiology. There are few tools to further these areas of study. Pancreatic duct infusion is an important technique that can allow for lineage tracing, gene introduction, and cell line-specific targeting. The technique requires the intricate dissection of the second portion of the duodenum and ampulla, followed by the occlusion of the bile duct and the cannulation of the pancreatic duct. Although the technique is technically challenging at first, the applications are myriad. Ambiguity in the specifics of the procedure between groups highlighted the need for a standard protocol. This work describes the expression of a green fluorescent protein (GFP) within the pancreas after the pancreatic duct infusion of a viral vector expressing GFP versus a sham surgery. The infusion and therefore expression is specific to the pancreas, without expression present in any other tissue type.

Pancreatic duct infusion is an important technique that can allow for lineage tracing, gene introduction, and cell line-specific targeting. A pancreatic duct infusion technique for drug and viral delivery to pancreatic cells is presented here.

Infusione del dotto pancreatico: un metodo efficace e selettivo di somministrazione di farmaci e virus

The pancreas is a bifunctional organ with both endocrine and exocrine components. A number of pathologies can afflict the pancreas, including diabetes, ...

Murine Salivary Functional Assessment via Pilocarpine Stimulation Following Fractionated Radiation

Hyposalivation is commonly observed in the autoimmune reaction of Sj&#246;gren's syndrome or following radiation injury to the major salivary glands. In these cases, questions remain regarding disease pathogenesis and effective interventions. An optimized technique that allows functional assessment of the salivary glands is invaluable for investigating exocrine gland biology, dysfunction, and therapeutics. Here, we present a step by step approach to performing pilocarpine stimulated saliva secretion, including tracheostomy and the dissection of the three major murine salivary glands. We also detail the appropriate murine head and neck anatomy accessed during these techniques. This approach is scalable, allowing for multiple mice to be processed simultaneously, thus improving the efficiency of the work flow. We aim to improve the reproducibility of these methods, each of which has further applications within the field. In addition to saliva collection, we discuss metrics for quantifying and normalizing functional capacity of these tissues. Representative data are included from submandibular glands with depressed salivary gland function 2 weeks following fractionated radiation (4 doses of 6.85 Gy).

We present a detailed approach to performing saliva collection, including murine tracheostomy and the isolation of three major salivary glands.

Valutazione funzionale salivare murina attraverso dopo stimolazione Pilocarpina frazionato radiazione

Hyposalivation is commonly observed in the autoimmune reaction of Sj&#246;gren's syndrome or following radiation injury to the major salivary glands. In ...

Radiation Treatment of Organotypic Cultures from Submandibular and Parotid Salivary Glands Models Key In Vivo Characteristics

Hyposalivation and xerostomia create chronic oral complications that decrease the quality of life in head and neck cancer patients who are treated with radiotherapy. Experimental approaches to understanding mechanisms of salivary gland dysfunction and restoration have focused on in vivo models, which are handicapped by an inability to systematically screen therapeutic candidates and efficiencies in transfection capability to manipulate specific genes. The purpose of this salivary gland organotypic culture protocol is to evaluate maximal time of culture viability and characterize cellular changes following ex vivo radiation treatment. We utilized immunofluorescent staining and confocal microscopy to determine when specific cell populations and markers are present during a 30-day culture period. In addition, cellular markers previously reported in in vivo radiation models are evaluated in cultures that are irradiated ex vivo. Moving forward, this method is an attractive platform for rapid ex vivo assessment of murine and human salivary gland tissue responses to therapeutic agents that improve salivary function.

Using three-dimensional organotypic cultures to visualize morphology and functional markers of salivary glands may provide novel insights into the mechanisms of tissue damage following radiation. Described here is a protocol to section, culture, irradiate, stain, and image 50&#8211;90 &#956;m thick salivary gland sections prior to and following exposure to ionizing radiation.

Trattamento radiante delle colture organotipiche da ghiandole salivari Sottomandibulari e parotidi modelli chiave in vivo caratteristiche

Hyposalivation and xerostomia create chronic oral complications that decrease the quality of life in head and neck cancer patients who are treated with ...

A Mouse Model for Chronic Pancreatitis via Bile Duct TNBS Infusion

Chronic pancreatitis (CP) is a complex disease involving pancreatic inflammation and fibrosis, glandular atrophy, abdominal pain and other symptoms. Several rodent models have been developed to study CP, of which the bile duct 2,4,6 -trinitrobenzene sulfonic acid (TNBS) infusion model replicates the features of neuropathic pain seen in CP. However, bile duct drug infusion in mice is technically challenging. This protocol demonstrates the procedure of bile duct TNBS infusion for generation of a CP mouse model. TNBS was infused into the pancreas through the ampulla of Vater in the duodenum. This protocol optimized drug volume, surgical techniques, and drug handling during the procedure. TNBS-treated mice showed features of CP as reflected by bodyweight and pancreas weight reductions, changes in pain-associated behaviors, and abnormal pancreatic morphology. With these improvements, mortality associated with TNBS injection was minimal. This procedure is not only critical in generating pancreatic disease models but is also useful in local pancreatic drug delivery.

Chronic pancreatitis (CP) is a disease characterized by inflammation and fibrosis of the pancreas, often associated with intractable abdominal pain. This article focuses on refining the technique to generate a mouse model of CP via bile duct infusion with 2,4,6 -trinitrobenzene sulfonic acid (TNBS).

Un modello murino per la pancreatite cronica tramite infusione di TNBS del dotto biliare

Chronic pancreatitis (CP) is a complex disease involving pancreatic inflammation and fibrosis, glandular atrophy, abdominal pain and other symptoms. ...