Name: retroductal submandibular gland instillation and localized fractionated irradiation in a rat model of salivary hypofunction
Uploaded: 2016-04-24T15:00:00.000+00:00
Duration: 7 min 44 s
Description: Watch this Scientific Journal Video about Retroductal Submandibular Gland Instillation and Localized Fractionated Irradiation in a Rat Model of Salivary Hypofunction at JoVE.com

We thank Dr. John Chang (Radiation Oncology, LSU Health Science Center) for assistance with radiation dose measurements. The study was supported by the American Cancer Society (Grant number: 116945-RSG-09-038-01-CCE), National Institute of Health (Grant number: R21CA173162) and the Feist-Weiller Cancer Center.

Tüm işlemler LSU Sağlık, Shreveport, Hayvan Bakımı ve Kullanımı Komitesi tarafından onaylanmış ve bakım ve laboratuvar hayvanlarının kullanımı için NIH kurallarına uygun olarak idi. Sıçan Submandibuler Tükürük Bezlerinin 1. kanülasyonu <ol><li> Şırınga-Boru Meclisi Hazırlanması <ol><li> bir neşter ile PE10 polietilen boru 10 cm uzunluğunda kesilir. işaret parmağı ve başparmak arasında boru iki ucunu tutun. nazik bir alev üzerinde boru orta bölümünü ısıtın ve yavaşça her iki tarafı çekerek uzunluğunu iki katına yumuşatılmış boru germek. </li><li> Konik bir ucu ile, her 2 - kanüller için 45 o lik bir açıda bir neşter ile orta hortumu kesilir. 0.5 cc İnsülin şırınga 29 G iğne üzerine takarak kanül olmayan konik ucunu genişletin. </li><li> kanül çıkarın ve enjektöre tuzlu su çözeltisi 220 ul çizin. Hava kabarcıklarını çıkarmak için hafifçe vurun. cannul Fitiğne üzerinde ve havayı dışarı atmak ve kanül yoluyla çözümün serbest akışını sağlamak için şırınga pistonu itin. 200 ul ses seviyesini ayarlayın. </li></ol></li><li> Submandibular Bez Duct içi Ağız kanülasyonu NOT: Otoklav araçlar prosedür öncesi ve sıcak bir boncuk sterilizatör prosedürlere arasında sterilize. <ol><li> Sprague Dawley sıçan tartılır ve bir hipodermik şırınga hesaplanan ketamin hacmi (42 mg / kg) / ksilazin (8 mg / kg) / asepromazin (1.4 mg / kg) karışımı dağıtın. </li><li> işaret parmağı ve bir elin başparmak arasındaki kuyruk tabanı tutup onu kavramak vücut üzerinde diğer yandan kaydırarak hayvan dizginlemek. Başparmak ve kalan parmakları ile gövde tutarken baş tarafı boyunca indeksi ve orta parmaklarınızı dinlenmek. </li><li> arka bacak kas anestezik enjekte edilir. ayak tutam ve palpebral refleks anestezi derinliği onaylayın. kuruluk süre önlemek için göz yağ sürünhayvan anestezi altında olduğunu. </li><li> Özel tasarlanmış bir platform (Şekil 1) hayvan koyun ve çapraz çubuk üst kesici dişlerin meşgul. alt ön dişlerin etrafına bir lastik bant döngü ve platform için demirleme ile alt çene aşağı çekin. </li><li> dil aracılığıyla steril dikiş geçmek ve ağız tabanını yükseltmek için yukarı kaldırın. Bir hemostatla sütürler Kelepçe ve çapraz çubuk üzerine geçirin. </li><li> Bir özel inşa edilmiş yanak yayıcı (Şekil 1) ile yanakları genişletin ve bir mikroskop altında ağız katta dilaltı papilla bulun. </li><li> hassas bir forseps kullanarak önceden PI10 boru konik ucunu tutun. Yavaşça dilaltı papilla üzerinde duktal deliğinin içine kanül ucu manipüle. o 3 geçirilerek kanül yerleştirme onaylamak - kanal içine 5 mm; herhangi bir engel olmadan geçer emin olun. </li></ol></li><li> Submandibuler Bezi Instillation <ol><li> Boynun ense deri altına atropin (0.5 mg / kg) enjekte edilir, ve tükürük salgılarında azaltılması için 10 dakika bekleyin. </li><li> siyanoakrilat (yapıştırıcı) bir damla kanal ağzına kanül Güvenli ve kurumasını bekleyin. yavaş 50 ml / dk&#39;lık bir oranda şırınga pistonunun basarak bezi (200 ul / bezi) &#39;de çözelti aşılamak. </li><li> Bir hemostatla boru Crush ve dikkatle şırınga çıkarın. Hayvan bilinç kavuşur kadar 60 dakika - 30 kanalda boru saklayın. dilini tutan dikiş çıkarın. </li><li> Ayrı bir kafesine hayvan aktarın ve kurtarma sırasında sıcak tutmak için bir ısı lambası kullanın. sternal recumbence korumak için kendine geldi kadar sahipsiz hayvan bırakmayın. </li><li> Hayvan tamamen ayaktan sonra, yiyecek ve su sınırsız erişim ile vivari adresinden ev. </li></ol></li></ol> 2. Lokalize fraksiyonlara IşınlamaTükürük Bezlerinin <ol><li> daha önce tarif edildiği gibi bir hayvan dizginlemek ve arka bacak kas içine ketamin (33 mg / kg) / ksilazin (6 mg / kg) / asepromazin (1 mg / kg) karışımı ile anestezi. anestezi derinliği onaylayın ve gözlere gres sürülmesi. </li><li> lineer hızlandırıcı masa üzerine hayvan sırtüstü yerleştirin ve kafa eğerek boynunu uzatmak. sternum üstüne mandibula alt sınırından bölgeyi kapsayacak şekilde radyasyon alanı (3 cm yarık genişliği) .birleştirecek. </li><li> bölge üzerinde 1 cm&#39;lik bir doku eşdeğer bolus yerleştirin ve radyasyon kaynağı ve 100 cm bolus tepesi arasındaki mesafeyi ayarlamak. </li><li> Bir lineer hızlandırıcı bir 6 MV foton ışını kullanarak hayvan (2.5 Gy) ışın tedavisi. pozlama süresini dikte edecek yüzeyi bolus radyasyon kaynağından oranı, alan büyüklüğü ve mesafesini Doz. ayarlamak akım hayvanlar, ~ 1 Gy / dak doz oranında ışınlanmıştır. </li><li> pozlama tekrarlayın; Toplam 8 2.5 Gy / güngün; 4 gün / arasında 2 gün ara ile bir hafta. kurtarma sırasında hayvan sıcak tutun. tamamen ayaktan sonra vivari hayvan aktarın. </li><li> Bezi fonksiyon 14 ölçmek için radyasyon sonrası uyarılan submandibuler bez tükürük 8 hafta toplayın. Soğuk% 4 paraformaldehid / fosfat tamponlu tuzlu su, pH 7.2, kardiyak perfüzyon ile anestezi altında hayvanlara öldürülür. Histolojik ve immünohistokimyasal 14 analizleri için submandibuler bezleri kökünü kazımak. </li></ol>

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The authors have nothing to disclose.

Tükürük bezleri sık sık baş boyun kanseri, boyun düğüm seçmeli ablasyon veya bölgesel hematolojik maligniteler için radyoterapi uygulanan hastalarda doku iyileşme eşiğin ötesinde radyasyon dozları alırsınız. bezinin sıvı salgılayan asinüs hücrelerinde terminal olarak farklılaşmış olsa da, paradoksal radyasyona hassas olan. salgı işlevi kronik düşük tükürük çıktı radyasyon ve geri dönüşümsüz bezi hasarı sonuçlarının ilk hafta içinde düşer. yoksul bezi fonksiyon ve ensue...

Sağlıklı dokularda kollateral yok kanser tedavilerinin zararlı yan etkilerin bir dizi oluşturur. radyasyon alanları ile yalan önemli tükürük bezlerinin bir kısmı veya tamamı kaçınılmaz imha edilir. Bu nedenle, kemik iliği transplantasyonu öncesinde baş ve boyun kanseri, servikal lenfoma, ya da tam vücut radyasyonu için radyoterapi uygulanan hastaların çoğunda radyasyon, tükürük bezi hipofonksiyon 2-6 en yaygın ve inatçı olumsuz etkileri birini çeker. tükürük bezlerinin sıvı üreten asiner hücreleri radyasyona akut duyarlıdır. tükürük bezlerinin hasar tükürük akışı gibi tükürük hipofonksiyonu ifade edilen bir durumun ciddi azalmasını neden olur. Tükürük akışındaki azalma kronik gibi çiğneme, yutma, konuşma ve tat, ama şiddetli ağrı, mukozal yırtık, yutma güçlüğü, fırsatçı enfeksiyonların hastalıklı sekel gibi temel sözlü faaliyetlerini bozar ve diş çürükleri bir kötüleşirHastanın refahı ve fonksiyon 2,3. radyoterapi ile ilişkili tükürük hücre kaybı geri döndürülemez olduğundan, ağız kuruluğu hiçbir düzeltici bir tedavisi yoktur. Yapay tükürük ikame ve prosecretory ilaçlarla belirtileri yatıştırılması odaklanan güncel tedavi uzun süreli rahatlama 6 etkisizdir. Geliştirilmiş Radyasyon uygulama teknikleri durumun şiddetini azaltma yardımcı olmasına rağmen, normal doku toksisitesi ve komplikasyonları, kanser tedavisinde 6,7 sınırlayıcı bir faktördür kalır. radyoterapi ile ilişkili komplikasyonları önlemek için önleyici tedbirler, bu nedenle norm haline gelmektedir. DNA onarımı serbest oksijen radikallerinin, bakıcı hücreleri oluşumunu temizlemek ya da geliştirmek Radyo koruyucu maddeler tükürük hipofonksiyona 8-11 önlemek için araştırılmaktadır. ekzokrin tükürük bezlerinin salgıları ana boşaltım kanalları yoluyla ağzına süzün. th Intra-Oral kanülasyonkontrast maddelerin enjeksiyonu için e boşaltım kanalları ayaktan prosedür olarak rutin olarak yapılır. Benzer bir yaklaşım kullanarak, tükürük bezleri doğrudan lokalize tedavi 12 hedeflenebilir. Dışında sistemik yan etki riskini azaltarak, retroductal bezi damlatma faydaları ekledi. duktal ağacının etrafında tükürük hücrelerin tek tabakalı düzenlemesi tüm tükürük epitel hücreleri hedef sağlar, ve bir engel olarak bezi eylemlerinin lifli kapsülleme istenmeyen terapötik yayılmasını azaltmak için. Özünde, tükürük bezleri gibi radyasyona bağlı tükürük hipofonksiyon olarak bezi rahatsızlıkları hedeflenen tedavi için optimal uygundur. haftalık bir süre için - (2.5 Gy / fraksiyon / gün, haftada beş gün 1.8), kanser tedavisi için geleneksel radyasyon küçük dozlarda verilir. Bu nedenle, deneysel modellerde uzun süren radyasyon düzeni karşı etkinliğini gösteren bir radyo-koruyucu tedavi daha klinik bir etkiye sahiptir. Comprofraksiyonlara radyasyondan sonra mised tükürük fonksiyonu küçük hayvanlar kaydedildi, ancak radyasyon kaynağı, doz fraksiyonu ve kullanılan protokoller çeşitli 9,10,13 vardır edilmiştir. Bu rapor retroductal teslimat ve hasta ilgili radyasyon kaynağı ve doz fraksiyonu kullanarak sıçan submandibuler bezleri lokalize radyasyon yöntemlerini belirler.

<table><tbody><tr><td>Intramedic Polyethylene tubing (PE10)</td><td>Becton Dickson</td><td>427401</td><td></td></tr><tr><td>1/2 cc Insulin Syringe U-100</td><td>Becton Dickson</td><td>309306</td><td></td></tr><tr><td>Artificial Tears</td><td>Miller Vet Supply&amp;nbsp;</td><td>5098-9840-64</td><td></td></tr><tr><td>Hot Bead Sterilizer</td><td>Fine Science Tools</td><td>18000-45</td><td></td></tr><tr><td>Perma-Hand silk suture</td><td>Ethicon</td><td>K833H</td><td></td></tr><tr><td>Graefe forcep</td><td>Fine Science Tools</td><td>11051-10</td><td></td></tr><tr><td>Olympus SZX16 Stereo Microscope</td><td>Hunt Optics and Imaging</td><td></td><td></td></tr><tr><td>6 MV Linear Accelerator</td><td>Elekta</td><td></td><td></td></tr><tr><td>Bolus - Skinless&amp;nbsp;</td><td>Civco</td><td>MTCB410</td><td></td></tr><tr><td>Heat Lamp</td><td>Braintree Scientific</td><td>HL-1&amp;nbsp;110V</td><td></td></tr></tbody></table>

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.

minimal invaziv sialografi tekniklerle, büyük tükürük bezlerinin lokal tedavi uygulanabilir. Sıçan submandibular tükürük bezlerinde Retroductal yönetim Wharton kanallarının ağız içi kanülasyon (Şekil 2) tarafından denendi. ağız katta bulunan dil-altı papilla açık Wharton tükürük kanalları, ancak orifisleri kolayca görünür değildir. Kateterin takıldığı, bu nedenle, yumuşak tarama yapıldı. kanül yerleştirerek şanssız kanama veya k...

Watch this Scientific Journal Video about Retroductal Submandibular Gland Instillation and Localized Fractionated Irradiation in a Rat Model of Salivary Hypofunction at JoVE.com

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

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.

Tükürük hipofonksiyonunun bir Sıçan Modelinde Retroductal Submandibuler Bezi damlatma ve yerelleştirilmiş fraksiyonlara Işınlama

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

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JoVE Journal

Medicine

11.3K Görüntüleme.  Louisiana State University Health Sciences Center. 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.

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

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

Severe salivary gland hypofunction is frequently found in patients with Sj&ouml;gren's syndrome and those who receiving therapeutic 
irradiation in their head and neck regions for cancer treatment. Both groups of patients experience symptoms such as xerostomia (dry mouth), dysphagia 
(impaired chewing and swallowing), severe dental caries, altered taste, oro-pharyngeal infections (candidiasis), mucositis, pain and discomfort.
One innovative approach of regenerative medicine for the treatment of salivary gland hypo-function is speculated in RS Redman, E Mezey 
et al. 2009: stem cells can be directly deposited by cannulation into the gland as a potent method in reviving the functions of the impaired organ. Presumably, 
the migrated foreign stem cells will differentiate into glandular cells to function as part of the host salivary gland. Also, this cannulation technique is an 
expedient and effective delivery method for clinical gene transfer application.
Here we illustrate the steps involved in performing the cannulation procedure on the mouse submandibular salivary gland via the Wharton's duct 
(Fig 1). C3H mice (Charles River, Montreal, QC, Canada) are used for this experiment, which have been kept under clean conventional conditions at 
the McGill University animal resource center. All experiments have been approved by the University Animal Care Committee and were in accordance with the 
guidelines of the Canadian Council on Animal Care.
For this experiment, a trypan blue solution is infused into the gland through the opening of the Wharton's duct using a 
insulin syringe with a 29-gauge needle encased inside a polyethylene tube. Subsequently, the mouse is dissected to show that the infusions migrated 
into the gland successfully.

A protocol for the cannulation of the mouse submandibular salivary gland via the Wharton's duct is described. For this experiment, the trypan blue solution is used as a dyer to demonstrate how this technique effectively delivers infusions into the targeted gland, and to suggest the reliability of this new approach as a potential clinical drug/cell therapy for the regeneration of salivary glands.

Wharton Kanalı üzerinden Fare Submandibular Tükürük Bezi kanülasyonu

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

Tıp

Isolation of Mouse Salivary Gland Stem Cells

Mature salivary glands of both human and mouse origin comprise a minimum of five cell types, each of which facilitates the production and excretion of saliva into the oral cavity. Serous and mucous acinar cells are the protein and mucous producing factories of the gland respectively, and represent the origin of saliva production. Once synthesised, the various enzymatic and other proteinaceous components of saliva are secreted through a series of ductal cells bearing epithelial-type morphology, until the eventual expulsion of the saliva through one major duct into the cavity of the mouth. The composition of saliva is also modified by the ductal cells during this process.
In the manifestation of diseases such as Sj&ouml;gren's syndrome, and in some clinical situations such as radiotherapy treatment for head and neck cancers, saliva production by the glands is dramatically reduced 1,2. The resulting xerostomia, a subjective feeling of dry mouth, affects not only the ability of the patient to swallow and speak, but also encourages the development of dental caries and can be socially debilitating for the sufferer.
The restoration of saliva production in the above-mentioned clinical conditions therefore represents an unmet clinical need, and as such several studies have demonstrated the regenerative capacity of the salivary glands 3-5. Further to the isolation of stem cell-like populations of cells from various tissues within the mouse and human bodies 6-8, we have shown using the described method that stem cells isolated from mouse salivary glands can be used to rescue saliva production in irradiated salivary glands 9,10. This discovery paves the way for the development of stem cell-based therapies for the treatment of xerostomic conditions in humans, and also for the exploration of the salivary gland as a microenvironment containing cells with multipotent self-renewing capabilities.

An optimized protocol for the isolation of stem cells from the mouse salivary gland is described. The method employs enzymatic and mechanical digestion, and permits isolation of salispheres containing cells with characteristics of stem cells.

Fare Tükürük Bezi Kök Hücre İzolasyonu

Mature salivary glands of both human and mouse origin comprise a minimum of five cell types, each of which facilitates the production and excretion of ...

Biyoloji

Retroductal Nanoparticle Injection to the Murine Submandibular Gland

Two common goals of salivary gland therapeutics are prevention and cure of tissue dysfunction following either autoimmune or radiation injury. By locally delivering bioactive compounds to the salivary glands, greater tissue concentrations can be safely achieved versus systemic administration. Furthermore, off target tissue effects from extra-glandular accumulation of material can be dramatically reduced. In this regard, retroductal injection is a widely used method for investigating both salivary gland biology and pathophysiology. Retroductal administration of growth factors, primary cells, adenoviral vectors, and small molecule drugs has been shown to support gland function in the setting of injury. We have previously shown the efficacy of a retroductally injected nanoparticle-siRNA strategy to maintain gland function following irradiation. Here, a highly effective and reproducible method to administer nanomaterials to the murine submandibular gland through Wharton's duct is detailed (Figure 1). We describe accessing the oral cavity and outline the steps necessary to cannulate Wharton's duct, with further observations serving as quality checks throughout the procedure.

Local drug delivery to the submandibular glands is of interest in understanding salivary gland biology and for the development of novel therapeutics. We present an updated and detailed retroductal injection protocol, designed to improve delivery accuracy and experimental reproducibility. The application presented herein is the delivery of polymeric nanoparticles.

Fare Submandibular bezi Retroductal nanopartikül enjeksiyon

Two common goals of salivary gland therapeutics are prevention and cure of tissue dysfunction following either autoimmune or radiation injury. By locally ...

Biyomühendislik

A Method for the Measurement of Salivary Gland Function in Mice

Patients with Sj&#246;gren&#39;s syndrome, an autoimmune disease affecting the exocrine glands, develop salivary gland inflammation and have reduced saliva production. Similarly, saliva production is severely compromised in patients receiving radiation treatment for head and neck cancers. Rodent models, developed to mimic these clinical conditions, facilitate an understanding of the disease pathogenesis and allow for the development of new therapeutic strategies. Therefore, the ability to accurately, reproducibly, and repeatedly measure salivary gland function in animal models is critical. Building on procedures previously described in the literature, a method was developed that meets these criteria and was used to evaluate salivary gland function in mice. An additional advantage of this new method is that it is easily mastered, and has little inter-operator variation. Salivary gland function is evaluated as the amount (weight or volume) or rate (mL/min) of saliva produced in response to pilocarpine stimulation. The collected saliva is a good source for the analyses of protein content, immunoglobulin concentrations, and other biomolecules.

Salivary gland hypofunction is a frequent consequence of autoimmune disease and radiation therapy. Reproducible evaluation of salivary gland function in mouse models of these diseases is a technical challenge. Here, a simple method for accurate and reproducible measurement of saliva production in mice is described.

Tükürük bezi işlevi farelerde ölçülmesi için bir yöntem

Patients with Sj&#246;gren&#39;s syndrome, an autoimmune disease affecting the exocrine glands, develop salivary gland inflammation and have reduced ...

İmmünoloji ve Enfeksiyon

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.

Dik Intravital mikroskopi için fare Submandibular Tükürük bezi hazırlanması

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

Ultrasound-guided Botulinum Toxin-A Injections: A Method of Treating Sialorrhea

Neurological diseases can be complicated by sialorrhea, an excessive flow of saliva. Patients suffering from moderate to severe sialorrhea have an impaired quality of life, often worsened by correlated complications such as aspiration pneumonia, oral infections, dental caries, and maceration of the skin. Diverse therapeutic approaches have been proposed for the treatment of sialorrhea, including surgery and the use of anticholinergic agents, with limited results and the possible occurrence of serious adverse events. Recently, botulinum toxin (BoNT) injection within the major salivary glands has been proposed in patients refractory to anticholinergic therapy, with the aim of inhibiting local acetylcholine release and gland activity. 
In order to obtain a better outcome in terms of reduction of saliva production, efficacy, duration, and avoidance of major adverse events, we developed an ultrasound-guided BoNT-type A injection technique accurately described in the text. Here we present a method of treating sialorrhea with bilateral parotid and submandibular gland BoNT-type A injections under ultrasound guidance. Four quadrants of the parotid gland and two quadrants of the submandibular gland are visualized and injected using two accesses and one access, respectively. 
The ultrasound-guided procedure provides a simple, non-invasive, real-time visualization of the muscular and glandular tissues and their surrounding structures, optimizing treatment efficacy and safety.

Botulinum toxin is injected into the salivary glands for the treatment of sialorrhea. The ultrasound-guided botulinum toxin injection in both the parotid and submandibular glands demonstrates a sustained efficacy and the absence of serious side effects.

Botulinum Toksin-A Enjeksiyonları Ultrason eşliğinde: Sialore Tedavisi A Yöntemi

Neurological diseases can be complicated by sialorrhea, an excessive flow of saliva. Patients suffering from moderate to severe sialorrhea have an ...

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.

Fare tükürük işlevsel değerlendirme Pilokarpin stimülasyon aşağıdaki yolu ile radyasyon şeker

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

Retrograde Parotid Gland Infusion through Stensen's Duct in a Non-Human Primate for Vectored Gene Delivery

Salivary glands are an attractive tissue target for gene therapy with promising results already leading to human trials. They are inherently capable of secreting proteins into the bloodstream and are easily accessible, making them potentially superior tissue sites for replacement hormone production or vaccination by gene transfer. Suggested methods for gene delivery include transcutaneous injection and retrograde infusion through salivary ducts. We demonstrate how to perform Retrograde Salivary Gland Infusion (RSGI) in non-human primates. We describe the important anatomic landmarks including identification of the parotid papilla, an atraumatic method of cannulating and sealing Stensen&#39;s Duct utilizing basic dental tools, polyethylene tubing, and cyanoacrylate, and the appropriate rate of infusion. While this is the least traumatic method of delivery, the method is still limited by the volume able to be delivered (&#60;0.5 mL) and the potential for trauma to the duct and gland. We demonstrate using fluoroscopy that an infusate can be fully delivered into the gland, and further demonstrate by immunohistochemistry the transduction of a typical vector and expression of the delivered gene.

Salivary glands have been proposed as a tissue target site for gene therapy, especially in the area of vaccination by gene transfer. We demonstrate gene delivery in a non-human primate model utilizing retrograde parotid infusion.

Vektörlü Gen Teslimatı için İnsan Olmayan Bir Primatta Stensen Kanalı aracılığıyla Retrograd Parotid Bezi İnfüzyonu

Salivary glands are an attractive tissue target for gene therapy with promising results already leading to human trials. They are inherently capable of ...

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.

Submandibular ve parotis tükürük bezlerinden Organotipik kültürlerin radyasyon tedavisi anahtar In vivo özellikleri

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

Fare Submandibular Bez Rejenerasyonunun Gerçek Zamanlı Canlı Görüntülenmesi

Interrogating Cell-Cell Interactions in the Salivary Gland via Ex Vivo Live Cell Imaging

Salivary gland regeneration is a complex process involving intricate interactions among various cell types. Recent studies have shed light on the pivotal role played by macrophages in driving the regenerative response. However, our understanding of this critical role has primarily relied on static views obtained from fixed tissue biopsies. To overcome this limitation and gain insights into these interactions in real time, this study outlines a comprehensive protocol for culturing salivary gland tissue ex vivo and capturing live images of cell migration.
The protocol involves several key steps: First, mouse submandibular salivary gland tissue is carefully sliced using a vibratome and then cultured at an air-liquid interface. These slices can be intentionally injured, for instance, through exposure to radiation, to induce cellular damage and trigger the regenerative response. To track specific cells of interest, they can be endogenously labeled, such as by utilizing salivary gland tissue collected from genetically modified mice where a particular protein is marked with green fluorescent protein (GFP). Alternatively, fluorescently-conjugated antibodies can be employed to stain cells expressing specific cell surface markers of interest. Once prepared, the salivary gland slices are subjected to live imaging using a high-content confocal imaging system over a duration of 12 h, with images captured at 15 min intervals. The resulting images are then compiled to create a movie, which can subsequently be analyzed to extract valuable cell behavior parameters. This innovative method provides researchers with a powerful tool to investigate and better understand macrophage interactions within the salivary gland following injury, thereby advancing our knowledge of the regenerative processes at play in this dynamic biological context.

Yazar Spotlight: Yaralanma Sonrası Tükürük Bezi Rejenerasyonunda Makrofaj-Epitel Hücre Etkileşimlerinin İncelenmesi 

Immunofluorescent imaging is constrained by the ability to observe complex, time-dependent biological processes in just a single snapshot in time. This study outlines a live-imaging approach conducted on precision-cut mouse submandibular gland slices. This approach allows for the real-time observation of cell-cell interactions during homeostasis and the processes of regeneration and repair.

Tükürük bezindeki hücre-hücre etkileşimlerinin ex vivo canlı hücre görüntüleme ile sorgulanması

Salivary gland regeneration is a complex process involving intricate interactions among various cell types. Recent studies have shed light on the pivotal ...