Incorporation of sentinel lymph node mapping in dogs with mast cell tumours: 20 consecutive procedures

The study hypothesis is that incorporation of sentinel lymph node (SLN) mapping in dogs presenting for mast cell tumour (MCT) removal would impact the recommended adjuvant therapy offered. Nineteen dogs were enrolled having either spontaneously occurring or incompletely excised MCTs. Staging included regional lymph node aspiration. SLN mapping was done with regional lymphoscintigraphy combined with intra‐operative lymphoscintigraphy and blue dye. Twenty MCTs in 19 dogs were excised with SLN mapping. Eight dogs had SLNs different from the closest node. Twelve dogs had metastasis in extirpated SLNs, seven occurred in MCTs with a MI ≤ 5. No correlation was noted between patient stage and the c‐KIT proto‐oncogene. Because of SLN staging, 8 of 19 dogs were offered additional therapy that would have otherwise been excluded. Anatomic sampling of lymph nodes in dogs with MCTs does not accurately reflect which lymph nodes are most likely to be receiving the draining tumour lymph.     

Sentinel lymph node mapping of the canine anal sac using lymphoscintigraphy: A pilot study

Sentinel lymph node mapping and biopsy are important parts of oncologic staging in human medicine. Sentinel lymph node mapping enables identification of the first lymph node to receive lymphatic drainage while avoiding unnecessary lymph node dissection. Anal sac adenocarcinoma is the most common malignant neoplasm of the canine perineal area. For dogs with anal sac adenocarcinoma, lympadenectomy and metastasis to the iliosacral lymphocentrum are negative prognostics indicators. The objectives of this prospective, two by two, crossover pilot study were to establish the feasibility of lymphoscintigraphy using Technetium‐99 sulfur colloid of the canine anal sac of healthy dogs, compare two injection techniques, and the time for identification of sentinel lymph nodes using each technique. We hypothesized that both intramural and perimural injections of the canine anal sac would identify similar sentinel lymph node drainage. The sentinel lymph node was identified in all dogs using either technique. Intramural injection of the canine anal sac showed radiopharmaceutical uptake faster than perimural injection technique (P = 0.040). There was concordance between intramual and perimural techniques for the sentinel lymph node identified in 50% of cases. A sacral lymph node was identified as sentinel in three of eight dogs (37.5%). Lymphoscintigraphy of the canine anal sac is safe and feasible in normal dogs; however, the method of injection technique seems to have a significant effect on the sentinel lymph node identified.     

Clinical usefulness of intradermal fluorescein and patent blue violet dyes for sentinel lymph node identification in dogs

The first lymph node receiving drainage from a specific anatomic region is referred to as the sentinel lymph node (SLN). This study sought to evaluate the intradermal use of two dyes, patent blue violet (PBV) and fluorescein (FL), for SLN mapping in the dog. Multiple intradermal injections were performed in five healthy dogs using two dyes, PBV in 0.9% NaCl and FL in solutions of 0.9% NaCl and 6% hetastarch. Skin flaps were raised and followed to the first area of discrete stain uptake. Areas of uptake were identified as lymph nodes grossly and by cytology. Identification of a SLN for each area of intradermal injection was accomplished for 98% of the injection sites. Intradermal injections of both PBV and FL dyes produce readily visible staining of lymphatic vessels and SLNs in healthy dogs and are sufficient to allow ready identification of these structures during postmortem dissection.     

Use of indirect lymphography to identify sentinel lymph node in dogs: a pilot study in 30 tumours

This study reports the clinical value of sentinel lymph node (SLN) mapping with indirect lymphography (IL) using iodized oil (IO) as a marker injected preoperatively around the site of the primary tumour and radiography or tomodensitometry for imaging. Surgical extirpation of the node was performed following peritumoural injection of methylene blue (MB). Twenty nine dogs affected by 30 palpable solid tumours were prospectively studied. SLNs were identified by IL in 96.6% of the IL studies. IL followed by MB studies were performed in 25 dogs (26 studies). In these studies, agreement between IL and MB was observed in 84.6%. One dog had a minor complication following IO injection. This protocol represents an attractive alternative to scintigraphy for SLN mapping. This less technically demanding protocol may provide a wider access to SLN identification for application in veterinary oncology.     

Indirect computed tomography lymphangiography with aqueous contrast for evaluation of sentinel lymph nodes in dogs with tumors of the head

Sentinel lymph node evaluation is widely used in human medicine to evaluate the first lymph node(s) to which a tumor drains. Sentinel lymph node biopsy allows avoidance of extensive lymphadenectomies in cases where the sentinel lymph node is negative for metastasis, thereby reducing patient morbidity. It has been shown that regional lymph nodes are not always the sentinel lymph node, thus identification and sampling of sentinel lymph nodes allows for more accurate staging, which is critical for treatment and prognostication in dogs with cancer. The objective of this prospective, pilot study was to determine if indirect computed tomography (CT) lymphangiography with aqueous contrast agent would successfully allow identification of sentinel lymph nodes in dogs with masses on the head. Eighteen dogs underwent CT lymphangiography. The sentinel lymph node was successfully identified within 3 min of contrast injection in 16 dogs (89%). Compression of lymphatic vessels from endotracheal tube ties and/or the patient's own body weight delayed or prevented identification of sentinel lymph nodes in two dogs (11%). Computed tomography lymphangiography with aqueous contrast can be used successfully to rapidly identify sentinel lymph nodes in dogs with masses on the head.     

Sentinel lymph node mapping by near‐infrared fluorescence imaging and contrast‐enhanced ultrasound in healthy dogs

Sentinel lymph node (SLN) mapping is a valuable and crucial diagnostic procedure in staging malignancies. We compared two non‐invasive techniques, near‐infrared (NIR) fluorescence imaging and contrast‐enhanced ultrasound (CEUS), to identify the SLNs in three superficial anatomical regions in an animal model. Six healthy laboratory dogs were included in a proof‐of‐concept trial. A NIR fluorescent dye (Indocyanine Green) and microbubbles (Sonovue) were consecutively injected subdermally in the Inguinal, axillary and popliteal region to map the SLNs. Transcutaneous NIR fluorescence imaging identified SLNs in 17 out of a total of 18 occasions. CEUS identified SLNs in all regions (18/18). Whereas NIR fluorescence imaging performed better in the visualization of the afferent lymphatic tract, CEUS demonstrated different filling patterns of the SLNs, a feature potentially critical for the concept of SLN mapping in cancer patients. Both NIR fluorescence imaging and CEUS are safe, non‐invasive, practical and accurate methods to perform real‐time transcutaneous SLN mapping with potential in a clinical setting.     

Pilot study to evaluate the efficacy of lymphotropic nanoparticle enhanced MRI for diagnosis of metastatic disease in canine head and neck tumours

This pilot study is designed to determine if lymphotropic nanoparticle enhanced MRI (LNMRI) is a viable technique for staging of naturally occurring canine malignant head and neck tumours. Previous imaging studies in veterinary medicine have shown variable sensitivity and specificity for determining metastasis for local lymph nodes in head and neck tumours. LNMRI utilizes ultra‐small superparamagnetic iron oxide nanoparticles (USPIOs) to help in the detection of metastatic disease in lymph nodes. USPIOs are phagocytized and localized to normal lymph nodes where they assist in evaluation for regions of effacement by cancerous cells. Six dogs underwent LNMRI for the diagnosis of metastatic lymph nodes. A truncated MRI consisting of transverse images of T2, T1 pre‐ and post‐contrast and T2* sequences were evaluated for presence of metastasis. Sentinel lymph nodes and lymph nodes with possible metastatic lesions were surgically excised for histological evaluation. In the initial phase of this study, 24 lymph nodes were included in analysis. Subjective observation by the primary investigator had a calculated sensitivity and specificity of 100% and 88% based on histological results. There were no negative side effects to the USPIOs noted in the limited number of patients in this study. Percentage signal intensity loss was calculated and found to be significantly different between metastatic and non‐metastatic lymph nodes (P‐value = .038). In conclusion, this pilot study shows that LNMRI has the potential to be a sensitive and specific method of diagnosing lymph node metastasis. Further research is warranted to determine if this method is clinically applicable and accurate.     

Sentinel lymph node mapping with computed tomography lymphography for mast cell tumours and a comparison between regional and sentinel lymph node histological status: Sixty-two cases

It is known that the regional lymph node (RLN) may not correspond to the sentinel lymph node (SLN) (the first lymph node draining the tumour), and many diagnostic techniques have recently been aimed at its detection. Although lymphoscintigraphy is the gold standard in both human and veterinary medicine for SLN mapping, it is relatively unavailable in veterinary medicine due to costs and difficult management of the radiotracer. This prospective study evaluated, as a first aim, the feasibility and sensitivity of the computed tomography lymphography (CTL) in detecting the SLN in 62 mast cell tumours (MCTs). The second aim was to evaluate the accuracy of the CTL in identifying the most representative lymph node of the patient's lymphatic status; the histological status of the SNL was compared with that of the RLN, to see in how many cases the patient's stage would have changed according to the RLN. When the RLN turned out to be also the SLN it was decided to excise, as a control LN, the one localised in the neighbourhood of the MCT (neighbouring lymph node; NLN). The detection rate was 90%, with failure of SLN identification in six cases. In 18 (32%) of 56 MCTs with a diagnostic CTL, the SLN did not correspond to the RLN. Forty-five MCTs were surgically removed, together with their corresponding SLN and RLN/NLN. Since the clinical stage of the patient would have changed in only 7% of cases, CTL is a reliable method of detecting the SLN and, for staging purposes, there is no need to remove other LNs.     

Canine oral mucosal mast cell tumours

Mast cell tumours (MCTs) are the most common cutaneous tumours of dogs, however rarely they can arise from the oral mucosa. This subset of MCT is reported to demonstrate a more aggressive clinical course than those tumours on the haired skin and the authors hypothesised that dogs with oral, mucosal MCT would have a high incidence of local lymph node metastasis at presentation and that this would be a negative prognostic factor. An additional hypothesis was that mitotic index (MI) would be prognostic. This retrospective study examines 33 dogs with MCTs arising from the oral mucosa. The results suggest that oral mucosal MCTs in the dog have a high incidence of lymph node metastasis at diagnosis (55%) which results in a poor prognosis. MI and nodal metastasis is highly prognostic. Loco‐regional progression is common in these patients and dogs with adequate local control of their tumour had an improved outcome. Despite a more aggressive clinical course, treatment can result in protracted survivals, even when metastasis is present.     

Patterns of nodal metastases,biological behaviour and prognosis of canine mast cell tumours of the pinna: A multi-institutional retrospective study

Canine cutaneous mast cell tumours (cMCTs) of the pinna have been associated with an aggressive biological behaviour, although data remain scarce. The knowledge acquired over the past years on histologic gradings, and the value of lymph node (LN) staging, may help in better characterizing this anatomical presentation. The first aim was to describe the frequency, location, and histologic appearance of LN metastases in cMCT of the pinna. A second aim was to evaluate prognosis. Medical records of dogs with cMCT of the pinna, that underwent tumour and sentinel (SLN) or regional LN (RLN) excision, were reviewed. The influence of potential prognostic variables on time to progression (TTP) and tumour-specific survival (TSS) was investigated. Thirty-nine dogs were included: 19 (48.7%) had Kiupel high-grade (K-HG) and 20 (51.3%) had low-grade (K-LG) MCTs. Eighteen (46.1%) dogs underwent SLN mapping: the superficial cervical LN was at least one of SLN in 17 (94.4%) cases. Twenty-two (56.4%) dogs had LN metastases; the superficial cervical LN was always involved. On multivariable analysis, only K-HG was associated with increased risk of progression (p = .043) and tumour-related death (p = .021). Median TTP and TSS were 270 and 370 days in K-HG, respectively; these were not reached in dogs with K-LG tumours (p < .01). cMCTs of the pinna are often K-HG and are also associated with a higher frequency of LN metastasis; however, we confirmed the independent prognostic value of histologic grading. A multimodal treatment may lead to favourable long-term outcome. Moreover, the superficial cervical LN is most often the SLN.     

Lymphography of the thoracic duct by percutaneous injection of iohexol into the popliteal lymph node of dogs: experimental study and clinical application

OBJECTIVE: To evaluate the efficacy of percutaneous administration of iohexol into the popliteal lymph node as a non-invasive technique for thoracic duct lymphangiography in dogs. STUDY DESIGN: Experimental study and clinical report. ANIMALS: Normal adult dogs (n=4) and 1 dog with recurrent chylothorax. METHODS: For the experimental study, 4 dogs (weight, 8.4-12.3 kg) had 5-10 mL iohexol injected percutaneously into 1 popliteal lymph node and then thoracic radiographs were taken. Popliteal lymph nodes were examined by histopathology 8 days later. One 25-kg dog with recurrent chylothorax had 25 mL iohexol injected into the right popliteal lymph node followed by thoracic radiography. RESULTS: In experimental dogs, the thoracic duct was best visualized on thoracic radiographs after administration of 10 mL iohexol. Clinically, no abnormalities were identified in the injected limb and except for 1 dog that had large numbers of siderocytes and erythrophagocytic macrophages in the injected lymph node, the histopathologic findings in the other injected popliteal lymph nodes were not different from contralateral nodes. In the clinical case, the thoracic duct was visualized, but there was leakage of iohexol around the node. CONCLUSION: The thoracic duct in dogs can be visualized by lymphography after percutaneous injection of iohexol (1 mL/kg at 2 mL/min) into the popliteal lymph node. CLINICAL RELEVANCE: Percutaneous popliteal lymph node administration of iohexol should be considered as an alternative to mesenteric lymph node injection for radiographic identification of the thoracic duct in dogs.     

The utility of staging in canine mast cell tumours

Current staging of canine mast cell tumours (MCTs) practiced by many veterinarians involves a minimum of lymph node (LN) assessment, abdominal ultrasound and thoracic radiography. Historically, some have advocated buffy coat and bone marrow evaluation. Two hundred and twenty dogs with MCT seen at a referral clinic were staged using LN palpation/cytology, thoracic radiography and abdominal ultrasound. The utility of each method was evaluated by considering prevalence of spread and future behaviour. At presentation, 30.9% of dogs had metastases to the local LN; 6.8% of all the dogs also had distant metastases. No dog had or developed distant metastasis in the absence of LN metastasis. No dog had convincing evidence of pulmonary metastasis. In this series, the local LN was sentinel to metastasis and in the absence of local LN metastasis, the utility of further staging was low. Thoracic radiography was not useful in the staging of canine MCT.     

Treatment of eight dogs with nasal tumours with alternating doses of doxorubicin and carboplatin in conjunction with oral piroxicam

OBJECTIVE: To determine the efficacy and toxicity of chemotherapy in the treatment of canine nasal tumours. DESIGN: Retrospective clinical study PROCEDURE: Eight dogs with histologically confirmed nasal tumours were staged by means of complete blood count, serum biochemical analysis, cytological analysis of fine needle aspirate of the regional lymph nodes, thoracic radiographs and computed tomography scan of the nasal cavity. All dogs were treated with alternating doses of doxorubicin, carboplatin and oral piroxicam. All dogs were monitored for side effects of chemotherapy and evaluated for response to treatment by computed tomography scan of the nasal cavity after the first four treatments. RESULTS: Complete remission was achieved in four dogs, partial remission occurred in two dogs and two had stable disease on the basis of computed tomography evaluation. There was resolution of clinical signs after one to two doses of chemotherapy in all dogs. CONCLUSIONS: This chemotherapy protocol was efficacious and well tolerated in this series of eight cases of canine nasal tumours.     

Lymph node staging of oral and maxillofacial neoplasms in 31 dogs and cats

A retrospective study was performed to report the histologic examination results of regional lymph nodes of dogs and cats with oral or maxillofacial neoplasms. Twenty-eight dogs and 3 cats were evaluated. Histologic examination results of standard and serial tissue sectioning of regional lymph nodes were recorded. When available, other clinical parameters including mandibular lymph node palpation, thoracic radiographs, and pre- and postoperative fine needle aspiration of lymph nodes were compared with the histologic results. Squamous cell carcinoma, fibrosarcoma, and melanoma were the most common neoplasms diagnosed in dogs. Squamous cell carcinoma and fibrosarcoma were diagnosed in cats. Of the palpably enlarged mandibular lymph nodes, 17.0% had metastatic disease histologically. Radiographically evident thoracic metastatic disease was present in 7.4% of cases. Preoperative cytologic evaluation of the mandibular lymph node based on fine needle aspiration concurred with the histologic results in 90.5% of lymph nodes examined. Postoperative cytologic evaluation of fine needle aspirates of regional lymph nodes concurred with the histologic results in 80.6% of lymph nodes examined. Only 54.5% of cases with metastatic disease to regional lymph nodes had metastasis that included the mandibular lymph node. Serial lymph node sectioning provided additional information or metastasis detection. Cytologic evaluation of the mandibular lymph node correlates positively with histology, however results may fail to indicate the presence of regional metastasis. Assessment of all regional lymph nodes in dogs and cats with oral or maxillofacial neoplasms will detect more metastatic disease than assessing the mandibular lymph node only.     

Association of tumour‐infiltrating regulatory T cells with adverse outcomes in dogs with malignant tumours

Regulatory T cells (Tregs) infiltrate into a variety of tumour tissues and associate with poor prognosis in humans. However, data on association of Treg infiltration with prognosis is limited in canine tumours. The purpose of this study was to examine the number of tumour‐infiltrating Tregs and its association with overall survival (OS) in dogs with malignant tumours. The following 168 canine tumours were included: 37 oral malignant melanomas (OMMs); 14 oral squamous cell carcinomas (OSCCs); 16 pulmonary adenocarcinomas (PAs); 37 mammary carcinomas (MCs); 36 mast cell tumours (MCTs) and 28 hepatocellular carcinomas (HCCs). Normal tissues were obtained from 8 healthy dogs as controls. The number of forkhead box P3 (Foxp3)‐positive Tregs in intratumoral and peritumoral areas was investigated by immunohistochemistry. OS was compared between high and low Treg groups. The number of intratumoral and peritumoral Foxp3‐positive Tregs was significantly higher in OMM, OSCC, PA and MC compared with each normal tissue. There were few Foxp3‐positive Tregs in MCT and HCC. With intratumoral Tregs, the OS in the high Treg group was significantly shorter than that in the low Treg group in OMM, OSCC and PA. With peritumoral Tregs, there was no significant difference for OS between the 2 groups in each tumour type. These results suggest that Tregs infiltrate into a variety of canine tumours and the abundance of Tregs are associated with poor prognosis in some solid tumour types.     

A review of sentinel lymph node evaluation and the need for its incorporation into veterinary oncology

Being the first lymph node or nodes to which many primary tumours reliably drain, the disease status of the sentinel lymph node/s (SLN) is important in the prediction of survival. SLN identification and biopsy are critical in the staging of human cancers. The status of the SLN helps determine prognosis and shape treatment plans. SLN evaluation is currently not routinely performed in veterinary oncology, not even at specialty oncology practices. Given the prognostic importance of lymph node involvement in tumours such as mammary gland carcinoma, osteosarcoma, synovial cell sarcoma and mast cell tumours, SLN evaluation should be incorporated into routine clinical practice so as to improve our clinical assessment of veterinary oncologic patients.     

Surgical complications following sentinel lymph node biopsy guided by γ-probe and methylene blue in 113 tumour-bearing dogs

Sentinel lymph node biopsy (SLNB) is an accepted veterinary surgical procedure given the impact of early detection of nodal metastases on staging of several canine malignancies. This study aims at reporting the incidence and risk factors for surgical complications of SLNB in tumour-bearing dogs. A total of 113 client-owned dogs that underwent tumour excision and SLNB guided by γ-probing and blue dye were retrospectively enrolled. Recorded variables included: signalment, location and number of extirpated lymphocenters and nodes, time for SLNB, histopathological status of excised nodes. Incidence of SLNB complications was calculated. They were classified as minor and major based on severity and required treatment, and as short-term (0–30 days) and long-term (31–90 days). Univariate analysis with generalized linear model with binomial error estimated the association between variables and incidence of SLNB complications. Significance was set at 5%. Median overall time for SLNB was 25 min. Surgeons excised one node in 38% of dogs and multiple nodes in 62% of cases, belonging to one (62%) or multiple (38%) lymphocenters. Metastases were detected in 45% of nodes. No intraoperative complications occurred. The overall incidence of postoperative complications of SLNB was 21,24%, the majority of which (91.67%) were minor. Only increasing dogs' weight was associated with an increased incidence of SLNB complications (p = .00976). Sentinel lymphadenectomy was associated with a relatively low incidence of complications, most of which were self-limiting. The low morbidity and previously reported impact on staging of SLNB justify its implementation to collect data for prognostic studies.     

Association of breed and histopathological grade in canine mast cell tumours

The aim of this study was to evaluate the relationship between breed and the histopathological grade of canine mast cell tumours (MCTs). A retrospective survey of pathology data of 9375 histopathologically confirmed diagnoses of cutaneous MCTs in the US was evaluated in the context of breed prevalence in over two million registered purebred dogs. Association of histopathological grade with breed, age, sex and spay/neuter status was assessed. The data indicate that the proportion of high‐grade tumours increases with advancing age, and that male and intact dogs have increased odds of developing high‐grade tumours. A significant difference in the proportion of high‐grade tumours between breeds was detected. The Pug was at significantly increased risk of developing low/intermediate‐grade tumours, but not high‐grade tumours, resulting in preponderance of less aggressive MCTs in this breed. The results of this study suggest a genetic association for the development of high‐grade MCTs.     

ASSESSMENT OF SENTINEL LYMPH NODE METASTASIS IN CANINE MAMMARY GLAND TUMORS USING COMPUTED TOMOGRAPHIC INDIRECT LYMPHOGRAPHY

In the staging process of the breast cancer, demonstrating metastasis of the sentinel lymph node (SLN) has an important prognostic value, in both humans and animals. The aim of this prospective case‐control study was to determine the diagnostic value of computed tomographic indirect lymphography (CT‐LG) for detecting SLN metastasis in dogs with mammary cancer. Thirty‐three female dogs with tumors in the abdominal and inguinal mammary glands were prospectively selected and subjected to CT‐LG, 1 and 5 min after injection of 1 ml of contrast agent (iopamidol) in the subareolar tissue of the neoplastic and the contralateral normal mammary glands. The pattern of postcontrast opacification, degree of postcontrast enhancement, and size and shape were assessed in 65 SLNs in total and were correlated with histopathological findings. The absence of opacification or heterogeneous opacification 1 min after contrast medium injection showed the highest sensitivity, specificity, and accuracy (93%, 100%, and 98.4%, respectively). In images taken 1 min after injection, an absolute density value lower than 444 Hounsfield units (HU) in the center of the SLN also provided significant sensitivity and specificity (93.8% and 75%, respectively). The size and shape of the SLN (maximum and minimum diameter, maximum/minimum diameter ratio, maximum diameter/height of fifth thoracic vertebral body ratio) showed the lowest sensitivity and specificity. Results of this study support the hypothesis that CT‐LG could help in the assessment of SLN metastasis in cases of mammary gland tumors in dogs.