Glycyrrhizic acid ameliorates submandibular gland oxidative stress, autophagy and vascular dysfunction in rat model of type 1 diabetes.

The burden of diabetes mellitus (DM) and associated complications is increasing worldwide, affecting many organ functionalities including submandibular glands (SMG). The present study aims to investigate the potential ameliorative effect of glycyrrhizic acid (GA) on diabetes-induced SMG damage. Experimental evaluation of GA treatment was conducted on a rat model of type I diabetes. Animals were assigned to three groups; control, diabetic and GA treated diabetic groups. After 8 weeks, the SMG was processed for assessment of oxidative stress markers, autophagy related proteins; LC3, Beclin-1 and P62, vascular regulator ET-1, aquaporins (AQPs 1.4 and 5), SIRT1 protein expressions in addition to LC3 and AQP5 mRNA expressions. Also, parenchymal structures of the SMG were examined. GA alleviated the diabetes-induced SMG damage via restoring the SMG levels of oxidative stress markers and ET-1 almost near to the normal levels most probably via regulation of SIRT1, AQPs and accordingly LC-3, P62 and Beclin-1levels. GA could be a promising candidate for the treatment of diabetes-induced SMG damage via regulating oxidative stress, autophagy and angiogenesis.

The mechanism of submandibular gland dysfunction after menopause may be associated with the ferroptosis.

Salivary gland dysfunction is a common symptom that occurs after menopause. This study was performed to investigate the mechanism of salivary gland dysfunction to confirm the relationship between ferroptosis and salivary gland dysfunction by ovariectomy. Forty-eight female rats were randomly divided into four groups (12 rats in each group). Histology, real time PCR, western blot, immunohistochemistry, electron microscopy, cytosolic iron assay, and salivary function were analyzed. Human salivary gland tissue analysis was also done. Lipogenesis and lipid deposition in the submandibular gland tissue occurred after ovariectomy. ROS generation, MDA+HAE was increased and GPX4 activity was decreased and in the OVX group compared to the CON group. Iron deposition in the submandibular gland tissue was increased in the OVX group. Submandibular gland fibrosis was increased and saliva secretion was decreased in the OVX group. In human submandibular gland analysis, lipid and iron deposition was also increased in the postmenopause group. This is the first in vivo study in which salivary gland dysfunction is associated with the ferroptosis in postmenopausal animal model. Increased lipid and iron deposition in normal submandibular gland tissues of postmenopausal women can suggest that the salivary gland dysfunction after menopause may be associated with the ferroptosis.

Switching to normal diet reverses kwashiorkor-induced salivary impairments via increased nitric oxide level and expression of aquaporin 5 in the submandibular glands of male Wistar rats.

Kwashiorkor, a form of malnutrition, has been shown to cause impaired salivary secretion. However, there is dearth of information on the mechanism that underlies this complication. Also, whether returning to normal diet after kwashiorkor will reverse these complications or not is yet to be discerned. Thus, this study aimed at assessing the mechanisms that underlie kwashiorkor-induced salivary impairments and to evaluate the effects of switching back to normal-diet on kwashiorkor-induced salivary impairments. Weaning rats were randomly divided into 3 groups (control group, kwashiorkor group (KG), re-fed kwashiorkor group (RKG)) of 7 rats each. The control group had standard rat chow while the KG and RKG were fed 2% protein diet for 6 weeks to induce kwashiorkor. The RKG had their diet changed to standard rat-chow for another 6 weeks. Blood and stimulated saliva samples were collected for the analysis of total protein, electrolytes, amylase, immunoglobulin A (IgA) secretion rate, leptin, and ghrelin. Tissue total protein, nitric oxide level, expressions of Na+/K+-ATPase, muscarinic (M3) receptor, and aquaporin 5 in the submandibular glands were also determined. Data were presented as means ± SEM and compared using ANOVA with Tukey's post hoc test. RKG showed improved salivary function evidenced by reduced salivary lag-time and potassium and increased flow rate, sodium, amylase, IgA secretion rate, leptin, submandibular nitric oxide level, and aquaporin 5 expression compared with KG. This study for the first time demonstrated that kwashiorkor caused significant reduction in salivary secretion through reduction of nitric oxide level and aquaporin 5 expression in submandibular salivary glands. Normal-diet re-feeding after kwashiorkor returned salivary secretion to normal.

Inducible nitric oxide synthase-mediated injury in a mouse model of acute salivary gland dysfunction.

AIM: Inducible nitric oxide synthase (iNOS) is a key regulator of the innate immune system. The aim of the current study was to explore whether innate immune-mediated iNOS and reactive nitrogen species acutely perturb acinar cell physiology and calcium homeostasis of exocrine salivary tissues. METHODS: Innate immunity in the submandibular gland of C57BL/6 mice was locally activated via intraductal retrograde infusion of polyinosinic:polycytidylic acid (poly (I:C). Expressions of iNOS and the activity of the reactive nitrogen species peroxynitrite, were evaluated by immunohistochemistry. Mice were pre-treated with the selective iNOS inhibitor aminoguanidine in order to substantiate the injurious effect of the nitrosative signal on the key calcium regulator sarcoplasmic/endoplasmic reticulum calcium ATPase (SERCA2b) and calcium signalling. RESULTS: Challenging salivary gland innate immunity with poly (I:C) prompted upregulated expression of iNOS and the generation of peroxynitrite. Inhibition of iNOS/peroxynitrite revealed the role played by upregulated nitrosative signalling in: dysregulated expression of SERCA2b, perturbed calcium homeostasis and loss of saliva secretion. CONCLUSION: iNOS mediates disruption of exocrine calcium signalling causing secretory dysfunction following activation of innate immunity in a novel salivary gland injury model.

Acute Submandibular Swelling Complicating Arteriography With Iodide Contrast: A Case Report and Literature Review.

Iodide mumps is an uncommon condition induced by iodide-containing contrast. We present the first reported case of iodide mumps in mainland China, which occurred after carotid artery intervention.The patient, a 65-year-old Chinese male, had a history of dizziness, hypertension, diabetes, and right arm weakness. He had no history of allergies and had never previously received iodide-containing contrast. The patient's kidney function and other laboratory findings were normal. He underwent stenting of the left internal carotid artery (LICA) opening and received approximately 250 mL of a nonionic contrast agent (ioversol). Approximately 5 hours after angioplasty, bilateral local swellings were noted near the mandible; the masses were moderately firm and nontender.Iodide mumps was diagnosed in the patient. Intravenous dexamethasone (10 mg) was administered. The submandibular glands had shrunk by 11 hours after angioplasty, and they gradually became softer. The mandibular salivary glands had completely recovered by 5 days after surgery.Iodide mumps represents a rare late reaction to iodine-containing contrast media. This condition can occur in any patient receiving any iodinated contrast agent and may recur upon repeated exposure, but self-resolution can be expected within 2 weeks. All clinicians who use contrast media or iodide should be aware of this condition.

Treatment of salivary gland hypofunction by transplantation with dental pulp cells.

OBJECTIVE: This study aimed to establish a mouse model in which dental pulp cells (DPCs) could be used as a cell source for the treatment of salivary gland hypofunction. DESIGN: DPCs were isolated from green fluorescent protein (GFP)-expressing mice and were differentiated into dental pulp endothelial cells (DPECs). DPEC behaviour was studied in vitro and in vivo to investigate their capacity to participate in neovascularisation. For in vivo assessment, a combination of DPECs and Matrigel was subcutaneously injected into nude mice. Two weeks after injection, Matrigel plugs were analysed for CD31 and GFP. Furthermore, both submandibular glands of the irradiated mice were injected with DPECs. Eight weeks after irradiation, the effect of DPECs on saliva secretion was evaluated by measuring amounts of saliva secretion. RESULTS: DPECs showed typical endothelial morphology, including a cobblestone appearance. RT-PCR analysis of DPECs showed positive expression of CD31, foetal liver kinase-1, vascular-endothelial-cadherin, vascular endothelial growth factor-A and von Willebrand factor. DPECs reorganised into tube-like structures on Matrigel after 24h in vitro. Positive merged staining for both CD31 and GFP was observed in the tube-like structures, representative of the injected DPECs. The average saliva flow rate in mice treated with DPECs was significantly higher than that observed in mice treated with PBS (P=0.0452). CONCLUSIONS: Our results show that radiation-induced salivary hypofunction is partially reverted following transplantation of DPECs. We established a mouse model in which DPCs could be used as a cell source for the treatment of salivary gland hypofunction.

Thyroid hemiagenesis, ectopic submandibular thyroid tissue, and apparent persistent subclinical thyrotoxicosis.

BACKGROUND: Ectopic thyroid tissue (ETT) is a rare embryological abnormality characterized by the occurrence of thyroid tissue in a site other than its usual location. Thyroid hemiagenesis (HA) is also a very rare abnormality in which one thyroid lobe does not develop properly during embryology. We report a patient with left HA, submandibular ETT, and persistent mildly suppressed serum thyrotropin (TSH). PATIENT: A 38-year-old female patient was admitted with complaints of sweating and palpitations. She had no symptoms of neck compression. Thyroid ultrasonography revealed heterogeneity and hypoechogenicity of the right lobe and absence of the left lobe, the latter being confirmed by computed tomography. There was no ETT in the neck. A thyroid Tc-99m pertechnetate scan demonstrated two distinct areas of radiotracer uptake, one in the right lobe of the thyroid gland and one in the right submandibular region and lobe, but no uptake in the left thyroid bed. The serum free triiodothyronine was 2.89 pg/mL (2.5-3.9 pg/mL), and the serum free thyroxine was 0.86 ng/dL (0.61-1.12 ng/mL). The serum TSH was 0.11 mIU/L (0.34-5.60 mIU/L). CONCLUSIONS: This may be the first reported patient with HA and submandibular ETT. The patient probably also had thyroiditis with mild intermittent thyrotoxicosis based on her suppressed TSH and ultrasonography imaging of the right thyroid lobe.

Salivary gland function 5 years after radioactive iodine ablation in patients with differentiated thyroid cancer: direct comparison of pre- and postablation scintigraphies and their relation to xerostomia symptoms.

BACKGROUND: Chronic sialadenitis is one of the most frequent chronic complications after radioactive iodine (RAI) therapy for thyroid cancer. To evaluate the long-term effects of RAI ablation on salivary gland function, we investigated scintigraphic changes in salivary glands by direct comparison of two salivary gland scintigraphies (SGSs) taken before and at 5 years after an RAI ablation. METHODS: SGS was performed just before RAI ablation (pre-SGS) and ∼5 years after RAI ablation (F/U SGS) in 213 subjects who underwent thyroidectomy for thyroid cancer. The uptake score (U score) was graded, and the ejection fraction (EF) was quantified for the parotid and submandibular glands at pre-SGS and F/U SGS. Changes in salivary gland function were graded as mild, moderate, or severe according to the differences in U score and EF between the two SGSs. Xerostomia was assessed and compared with the SGS findings. RESULTS: Worsening of the U score was observed in 182 of 852 salivary glands (total: 21.3%; mild: 4.2%, moderate: 7.4%, severe: 9.7%), and 47.4% of the patients had a worsening U score for at least one of four salivary glands. A decrease in EF was observed in 173 of 852 salivary glands (total: 20.3%; mild: 5.4%, moderate: 6.8%, severe: 8.1%), and 43.7% of the patients experienced a decrease in the EF of at least one of the four salivary glands. Bilateral parotid gland dysfunction was the most commonly observed condition. Thirty-five (16.4%) patients complained of xerostomia at 5 years after RAI ablation. Scintigraphic changes in salivary gland function and xerostomia were more common in patients receiving 5.55 GBq, compared with 3.7 GBq. Xerostomia was more common in patients with submandibular gland dysfunction than those with parotid gland dysfunction (68.8% vs. 33.3%, p<0.05). The number of dysfunctional salivary glands was correlated with xerostomia (p<0.01). CONCLUSION: About 20% of the salivary glands were dysfunctional on SGS 5 years after a single RAI ablation, especially in patients who received higher doses of RAI. While parotid glands are more susceptible to (131)I-related damage, xerostomia was more associated with submandibular gland dysfunction and the prevalence of dysfunctional salivary glands.

Loss of PKCδ results in characteristics of Sjögren's syndrome including salivary gland dysfunction.

OBJECTIVES: Chronic infiltration of lymphocytes into the salivary and lacrimal glands of patients with Sjögren's syndrome (SS) leads to destruction of acinar cells and loss of exocrine function. Protein kinase C-delta (PKCδ) is known to play a critical role in B-cell maintenance. Mice in which the PKCδ gene has been disrupted have a loss of B-cell tolerance, multiple organ lymphocytic infiltration, and altered apoptosis. To determine whether PKCδ contributes to the pathogenesis of SS, we quantified changes in indicators of SS in PKCδ-/- mice as a function of age. Salivary gland histology, function, the presence of autoantibodies, and cytokine expression were examined. MATERIALS AND METHODS: Submandibular glands were examined for the presence of lymphocytic infiltrates, and the type of infiltrating lymphocyte and cytokine deposition was evaluated by immunohistochemistry. Serum samples were tested by autoantibody screening, which was graded by its staining pattern and intensity. Salivary gland function was determined by saliva collection at various ages. RESULTS: PKCδ-/- mice have reduced salivary gland function, B220+ B-cell infiltration, anti-nuclear antibody production, and elevated IFN-γ in the salivary glands as compared to PKCδ+/+ littermates. CONCLUSIONS: PKCδ-/- mice have exocrine gland tissue damage indicative of a SS-like phenotype.

Acute salivary gland hypofunction in the duct ligation model in the absence of inflammation.

OBJECTIVE: The commonly associated aetiology of salivary gland inflammation and salivary hypofunction has led to the widely held belief that inflammation causes salivary gland hypofunction. Indeed, our own recent study seemed to support this contention. Here, we tested the hypothesis that, in an acute duct ligation model, eliminating inflammation the submandibular gland would recover normal function. MATERIALS AND METHODS: Ligation of the rat submandibular gland excretory duct for 24 h was used to induce inflammation and salivary gland hypofunction. A group of duct ligated rats was compared with a second group given dexamethasone, on the day of duct ligation. Twenty-four hours later salivary gland function was assessed and salivary glands were collected. RESULTS: Histology and myeloperoxidase activity assay revealed a profound decrease in inflammatory cell infiltration of ligated glands from rats given dexamethasone, compared with ligated glands in the absence of dexamethasone. Salivary flow rate evoked by methacholine was decreased (P < 0.01) by approximately 56% (ligated vs control, 79 +/- 9 microl min(-1) g(-1)vs 177 +/- 11 microl min(-1) g(-1)) and salivary flow from ligated dexamethasone-treated and ligated glands was similar. CONCLUSION: Despite eliminating the inflammatory reaction in the ligated gland, salivary hypofunction was not reversed, suggesting that other mechanisms must be at work in the ligation-induced salivary hypofunction.

Severe life threatening maxillofacial infection in pregnancy presented as Ludwig's angina.

BACKGROUND: Ludwig's angina is a rapidly spreading cellulitis that may produce upper airway obstruction often leading to death. There is very little published information regarding this condition in the pregnant patient. CASE: A 24-year old black female was admitted at 26 weeks gestation with tooth pain, submandibular swelling, severe trismus, and dysphagea, consistent with Ludwig's angina. Her treatment included emergent tracheostomy, incision and drainage of associated spaces, teeth extraction, and antibiotic therapy. CONCLUSIONS: During a life threatening infectious situation such as the one described, risks of maternal and fetal morbidity include both septicemia and asphyxia. Furthermore, the healthcare provider must consider the risks that the condition and the possible treatments may cause the mother and her unborn child.

Recovery of rat submandibular salivary gland function following removal of obstruction: a sialometrical and sialochemical study.

Functional recovery of the rat submandibular gland following ligation of the main excretory duct was examined. Rat submandibular glands were ligated for 1, 4 and 8 weeks using a micro-clip with a plastic tube. Micro-clips were removed and glands were allowed to recover for periods of 8, 16 and 24 weeks. Submandibular glands were stimulated with autonomimetic drugs (methacholine and isoprenaline) and salivas were collected from atrophic or de-ligated and contralateral control glands. Glands recovered almost full size (92% of control gland) following 24 weeks of de-ligation. Saliva volume secreted by ligated/de-ligated (RSM) and control (LSM) glands were similar with different doses of agonists. Protein output expressed per gram of tissue wet weight was similar from both ligated/de-ligated and control glands with all doses of agonist. Sodium and chloride levels were higher from de-ligated glands than contralateral control glands. Protein electrophoresis showed similar profiles of salivary proteins in all samples with some minor differences. Acinar cells in de-ligated glands showed a normal morphology, as indicated by light microscopy, whilst granular ductal cells were fewer and contained fewer secretory granules. Sodium potassium ATPase staining of striated ducts in de-ligated glands was similar to that of control glands. It can be concluded that rat submandibular glands can regenerate following severe atrophy and secrete normal amounts of saliva containing broadly a full profile of secretory proteins. In contrast to acinar cells, ductal cells appear not to recover full function.

Glandular function after intraoral removal of salivary calculi from the hilum of the submandibular gland.

We studied 43 patients (25 women and 18 men) who had salivary calculi removed from the hilum of the submandibular gland. Preoperatively they had clinical and radiographic examinations, and glandular function was measured scintigraphically in 38 patients. Postoperative follow-up was based on history, clinical examination, structured questionnaire, and scintigraphy. Stone(s) were removed successfully in 42 patients (97%). During the follow-up of a mean of 24 months (range 4-47), 37 patients were symptom-free and 2 patients had mild obstructive symptoms that did not require intervention. The other 4 patients had repeated infections that necessitated removal of the gland under general anaesthesia. Preoperative and postoperative scintigraphic assessments were made in 37 patients (88%). There was a significant increase in the functional fraction and the excretion rate in the gland after removal of the calculus. We conclude that glandular function improves to varying degrees in most patients after the removal of a salivary calculus.

An unusual cause of obstructive sialadenitis.

A case of bilateral recurrent submandibular sialadenitis is presented in a patient with decreased salivary flow. The condition was caused by trapping of the submandibular papillae between the lingual flange of the denture and the enlarged genial tubercles. Removal of these causes led to resolution of her symptoms. The value of plain radiography, sialography and scintigraphy is discussed.

Radiation-induced progressive decrease in fluid secretion in rat submandibular glands is related to decreased acinar volume and not impaired calcium signaling.

The mechanism(s) of radiation-induced salivary gland dysfunction is poorly understood. In the present study, we have assessed the secretory function (muscarinic agonist-stimulated saliva flow, intracellular calcium mobilization, Na+/K+/2Cl- cotransport activity) in rat submandibular glands 12 months postirradiation (single dose, 10 Gy). The morphological status of glands from control and irradiated rats was also determined. Pilocarpine-stimulated salivary flow was decreased by 67% at 12 months (but not at 3 months) after irradiation. This was associated with a 47% decrease in the wet weight of the irradiated glands. Histological and morphometric analysis demonstrated that acinar cells were smaller and occupied relatively less volume and convoluted granular tubules were smaller but occupied the same relative volume, while intercalated and striated ducts maintained their size but occupied a greater relative volume in submandibular glands from irradiated compared to control animals. In addition, no inflammation or fibrosis was observed in the irradiated tissues. Carbachol- or thapsigargin-stimulated mobilization of Ca2+ was similar in dispersed submandibular gland cells from control and irradiated animals. Further, [Ca2+]i imaging of individual ducts and acini from control and irradiated groups showed, for the first time, that mobilization of Ca2+ in either cell type was not altered by the radiation treatment. The carbachol-stimulated, bumetanide-sensitive component of the Na+/K+/ 2Cl- cotransport activity was also similar in submandibular gland cells from control and irradiated animals. These data demonstrate that a single dose of gamma radiation induces a progressive loss of submandibular gland tissue and function. This loss of salivary flow is not due to chronic inflammation or fibrosis of the gland or an alteration in the neurotransmitter signaling mechanism in the acinar or ductal cells. The radiation-induced decrease in fluid secretion appears to be related to a change in either the water-handling capacity of the acini or the number of acinar cells in the gland.

[Evaluation of normal submandibular gland function by sialo-scintigraphy].

OBJECTIVE: To study the normal glandular function of the submandibular gland as a background for evaluation of changes in submandibular gland diseases. METHODS: The backgrounds of both submandibular area and temporal area were analyzed comparatively with Emission Computerized Tomography in eleven pairs of normal submandibular gland. RESULTS: About 2.18 times of the temporal background equals to submandibular area background; the Concentrate Index of normal submandibular gland was 1.42 +/- 0.89; the Secretory Index was (109.4 +/- 39.8)%, the Secretory Index Ratio and the Concentrate Index Ratio were (77.2 +/- 17.2)% and 71.0 +/- 15.2% respectively; and the Function Index was (89.9 +/- 7.4)%. CONCLUSION: The use of submandibular area background and functional index to represent submandibular gland function is more accurate and more sensitive. Normal functional index can provide the basis for researching submandibular gland disease.

Adrenergic receptors and secretory responses of the rat submandibular salivary gland after periodic incisor reduction.

The lower and upper incisors of female rats were repeatedly reduced every 48 hr for 21 days. A marked enlargement of the submandibular glands was observed at the end of this period. One day after the final reduction, dose dependent curves to phenylephrine and isoproterenol were obtained in relation to salivary flow rates. Secretory responses, expressed as mg/gland, showed that the dose response curve to the alpha1-adrenomimetic drug was not modified by treatment while that for isoproterenol was shifted to the right of the control. When the responses were expressed as microg of saliva/mg of wet tissue, the dose-response curve to both agonists was shifted to the right in the incisor-reduced group. (Activation of alpha2-adrenergic receptors by clonidine did not inhibit the responses to phenylephrine in the incisor-reduced rats.) Radioligand binding assays of alpha1-, beta- and alpha2-receptors did not show differences between control and experimental glands in terms of densities (Bmax) or affinities (Kd). The lack of correlation between the decrease in alpha2- and beta-mediated responses and the radioligand bindings suggests that postreceptor mechanisms are involved in the diminished secretory responses of the rat submandibular gland after periodic reduction or amputation of incisors.

[A new therapeutic approach to salivary calculi: extracorporeal lithotripsy]. / Nouvelle approche thérapeutique des calculs salivaires: la lithotrypsie extra corporelle.

A common pathology of the salivary gland, salivary lithiases are usually removed by open surgery. Most frequently localized in the sub-mandibular gland, calculi can also occur in the parotid gland. We developed a new procedure to treat salivary lithiasis using extracorporeal electromagnetic shock waves. New material was especially adapted for the salivary gland for lithotripsy without anesthesia. Two-hundred patients were treated; 63% had not lithiasis after treatment and 37% had small residual stones measuring less than 3 mm; 97% of the patients were symptom-free and salivary excretion was improved. We believe that extracorporeal electromagnetic shock waves is a safe noninvasive and well tolerated alternative to open surgery. We use lithotripsy as first line therapy for salivary lithiasis.

Parotid and submandibular sialadenitis treated by salivary gland excision.

BACKGROUND: The purpose of the present study was to compare and contrast the clinicopathological features and treatment outcome of chronic sialadenitis involving the parotid and submandibular glands, among patients who were treated by excision of the affected gland. METHODS: In a series of 88 patients treated over an 8-year period, 47 had parotid sialadenitis and 41 had submandibular sialadenitis. In the parotid group, 63% of patients were symptomatic for longer than 6 months compared with 27% in the submandibular group. Calculi were implicated in the disease process in 24% of patients with parotid sialadenitis, compared with 73% of patients with submandibular disease. RESULTS: Patients with parotid sialadenitis had superficial (n = 14) or near-total parotidectomy (n = 35), while those with submandibular sialadenitis underwent total gland excision. Complications occurred twice as frequently in the parotidectomy group. The rate of temporary facial nerve weakness was 29% after parotidectomy while marginal mandibular nerve dysfunction occurred after 12% of submandibular excisions. Both procedures were highly effective in permanently relieving the symptoms of sialadenitis. CONCLUSIONS: Parotid sialadenitis is infrequently associated with stones and tends to run a longer course before surgical intervention is necessary. Submandibular sialadenitis usually presents earlier, is secondary to calculi and requires early intervention.

[The relationship of function and pathology of submandibular gland with sialolithiasis].

Submandibular glands with sialolithiasis are analysed histopathologically. The formation of the calculi of the submandibular glands may be related to the sialoadentitis and to the changes of saliva composition. Mineralization of the organic matrix may be one of the cause of calculus formation. By comparison of the glandular function with the pathology of the gland, it shows that the reduction of glandular function is associated with the severity of grandular inflammation. When function index is normal, grandular inflammation is in the Stage I or II, while it decreases or zero, grandular inflammation is in the Stage III or IV. Glandular function index may predict the severity of grandular inflammation.