Cancer-associated fibroblasts promote bone invasion in oral squamous cell carcinoma

Background: The molecular mechanisms involved in the invasion of bone by oral squamous cell carcinomas (OSCC) are poorly understood, and little is known about the role of cancer-associated fibroblasts (CAF), the presence of which confers a poor prognosis. Methods: Clinicopathological data from 277 OSCC cases involving bone resections were reviewed, and 32 cases thoroughly analysed histologically. Immunohistochemistry was used to examine αSMA, RANKL and OPG. Western blotting and qPCR were used to assess myofibroblast (CAF-like) differentiation, RANKL and OPG expression in vitro, and RANKL secretion was analysed by ELISA. Osteoclastogenesis was examined using TRAP staining, multinucleation and pit forming assays. Results: Fibrous stroma intervened between tumour and bone in the majority of cases, with no direct contact between cancer cells and bone. RANKL and OPG, two proteins key to regulating bone resorption, were expressed in tumour cells as well as fibrous stroma adjacent to bone and αSMA-positive myofibroblastic CAF were consistently seen infiltrating into bone ahead of tumour cells. Human primary osteoblasts cultured with conditioned media from human OSCC-derived cells and human primary CAF showed a significant increase in RANKL and a decline in OPG mRNA expression. RANKL secretion was significantly increased in primary oral fibroblasts induced to differentiate into a CAF-like phenotype by transforming growth factor-β1 (TGF-β1) treatment and in primary CAF. Indirect co-culture of murine macrophages with conditioned media from CAF (experimentally derived and isolated from OSCCs) resulted in a marked increase in osteoclastogenesis (in excess of that provoked by cancer cells) determined by tartrate-resistant acid phosphatase activity, multinucleation and resorption pit formation. Conclusions: This study is the first to describe a functional role for CAFs in bone invasion and turnover, identifying a novel potential therapeutic target and diagnostic indicator in this difficult to treat bone invasive malignancy

Hypoglossal nerve trunk stimulation: electromyography findings during drug-induced sleep endoscopy: a case report

Abstract Background Literature has demonstrated hypoglossal nerve stimulation to be a safe and effective treatment for patients with obstructive sleep apnea nonadherent to positive airway pressure therapy. However, the recommended criteria for patient selection are still unable to identify all the unresponsive patients, highlighting the need for improved understanding about hypoglossal nerve stimulation for obstructive sleep apnea. Case presentation A 48-year-old Caucasian male patient with obstructive sleep apnea had been successfully treated with electrical stimulation of the hypoglossal nerve trunk, documented by level 1 polysomnography data. However, due to snoring complaints, he underwent postoperation drug-induced sleep endoscopy for evaluation of electrode activation during upper airway collapse, aiming to improve electrostimulation parameters. Concurrent surface electromyography of the suprahyoid muscles and masseter was obtained. Activation of electrodes 2, 3, and 6 promoted upper airway opening most strongly at the velopharynx and tongue base during drug-induced sleep endoscopy. The same channels also significantly increased the electrical activity on suprahyoid muscles bilaterally, but predominantly on the stimulated side (right). The masseters also presented a considerable asymmetry in electrical potential on the right side (> 55%). Conclusion Beyond the genioglossus muscle, our findings demonstrate recruitment of other muscles during hypoglossal nerve stimulation, which may be attributed to the electrical stimulation of the nerve trunk. This data provides new insights on how stimulation of the hypoglossal nerve trunk may contribute to obstructive sleep apnea treatment

Upper airway collapsibility is associated with obesity and hyoid position.

Study objectivesUpper airway anatomy plays a major role in obstructive sleep apnea (OSA) pathogenesis. An inferiorly displaced hyoid as measured by the mandibular plane to hyoid distance (MPH) has been consistently associated with OSA. The hyoid is also a common landmark for pharyngeal length, upper airway volume, and tongue base. Tongue dimensions, pharyngeal length, and obesity are associated with OSA severity, although the link between these anatomical variables and pharyngeal collapsibility is less well known. We hypothesized that obesity as measured by body mass index (BMI), neck and waist circumferences, and variables associated with hyoid position (pharyngeal length, upper airway volume, and tongue dimensions) would be associated with passive pharyngeal critical closing pressure (Pcrit).DesignCross-sectional.SettingAcademic hospital.Patients34 Japanese-Brazilian males age 21 to 70 y.InterventionsN/A.Measurements and resultsWe performed computed tomography scans of the upper airway, overnight polysomnography, and Pcrit measurements in all subjects. On average, subjects were overweight (BMI = 28 ± 4 kg/m(2)) and OSA was moderately severe (apnea-hypopnea index = 29 [13-51], range 1-90 events/h). Factor analysis identified two factors among the studied variables: obesity (extracted from BMI, neck and waist circumferences) and hyoid position (MPH, pharyngeal length, tongue length, tongue volume, and upper airway volume). Both obesity and hyoid position correlated with Pcrit (r = 0.470 and 0.630, respectively) (P < 0.01). In addition, tongue volume, tongue length, pharyngeal length, and MPH correlated with waist and neck circumferences (P < 0.05).ConclusionsPharyngeal critical closing pressure is associated with obesity and hyoid position. Tongue dimensions, pharyngeal length, and the mandibular plane to hyoid distance are associated with obesity variables. These findings provide novel insight into the potential factors mediating upper airway collapse in obstructive sleep apnea

Different Craniofacial Characteristics Predict Upper Airway Collapsibility in Japanese-Brazilian and White Men.

BackgroundOSA pathogenesis is complex and may vary according to ethnicity. The anatomic component predisposing to OSA is the result of the interaction between bony structure and upper airway soft tissues and can be assessed using passive critical closing pressure (Pcrit). We hypothesized that Japanese-Brazilians and whites present different predictors of upper airway collapsibility, suggesting different causal pathways to developing OSA in these two groups.MethodsMale Japanese-Brazilians (n = 39) and whites (n = 39) matched for age and OSA severity were evaluated by full polysomnography, Pcrit, and upper airway and abdomen CT scans for determination of upper airway anatomy and abdominal fat, respectively.ResultsPcrit was similar between the Japanese-Brazilians and the whites (-1.0 ± 3.3 cm H2O vs -0.4 ± 3.1 cm H2O, P = .325). The Japanese-Brazilians presented smaller upper airway bony dimensions (cranial base, maxillary, and mandibular lengths), whereas the whites presented larger upper airway soft tissue (tongue length and volume) and a greater imbalance between tongue and mandible (tongue/mandibular volume ratio). The cranial base angle was associated with Pcrit only among the Japanese-Brazilians (r = -0.535, P < .01). The tongue/mandibular volume ratio was associated with Pcrit only among the whites (r = 0.460, P < .01). Obesity-related variables (visceral fat, BMI, and neck and waist circumferences) showed a similar correlation with Pcrit in the Japanese-Brazilians and the whites.ConclusionsJapanese-Brazilians and whites present different predictors of upper airway collapsibility. Although craniofacial bony restriction influenced Pcrit only in the Japanese-Brazilians, an anatomic imbalance between tongue and mandible volume influenced Pcrit among the whites. These findings may have therapeutic implications regarding how to improve the anatomic predisposition to OSA across ethnicities