Sublingual Tissue Research Articles

Microvascular Phenotyping in the Maastricht Study: Design and Main Findings, 2010-2018.

Microvascular dysfunction (MVD) is a common pathophysiological change that occurs in various diseases, such as type 2 diabetes mellitus (T2DM), heart failure, dementia, and depression. Recent technical advances have enabled noninvasive measurement and quantification of microvascular changes in humans. In this paper, we describe the protocols of the microvascular measurements applied in the Maastricht Study, an ongoing prospective, population-based cohort study of persons aged 40–75 years being carried out in the southern part of the Netherlands (baseline data assessment, November 2010–January 2020). The study includes a variety of noninvasive measurements in skin, retina, brain, and sublingual tissue, as well as plasma and urine biomarker assessments. Following this, we summarize our main findings involving these microvascular measurements through the end of 2018. Finally, we provide a brief perspective on future microvascular investigations within the framework of the Maastricht Study.

Association between sublingual microcirculation, tissue perfusion and organ failure in major trauma: A subgroup analysis of a prospective observational study.

IntroductionPrevious studies described impaired microvascular perfusion and tissue oxygenation as reliable predictors of Multiple Organ Failure in major trauma. However, this relationship has been incompletely investigated. The objective of this analysis is to further evaluate the association between organ dysfunction and microcirculation after trauma.Materials and methodsThis is a retrospective subgroup analysis on 28 trauma patients enrolled for the Microcirculation DAIly MONitoring in critically ill patients study (NCT 02649088). Patients were divided in two groups according with their Sequential Organ Failure Assessment (SOFA) score at day 4. At admission and every 24 hours, the sublingual microcirculation was evaluated with Sidestream Darkfield Imaging (SDF) and peripheral tissue perfusion was assessed with Near Infrared Spectroscopy (NIRS) and Vascular Occlusion Test (VOT). Simultaneously, hemodynamic, clinical/laboratory parameters and main organ supports were collected.ResultsMedian SOFA score at Day 4 was 6.5. Accordingly, patients were divided in two groups: D4-SOFA ≤6.5 and D4-SOFA >6.5. The Length of Stay in Intensive Care was significantly higher in patients with D4-SOFA>6.5 compared to D4-SOFA≤6.5 (p = 0.013). Total Vessel Density of small vessels was significantly lower in patients with high D4-SOFA score at Day 1 (p = 0.002) and Day 2 (p = 0.006) after admission; the Perfused Vessel Density was lower in patients with high D4-SOFA score at Day 1 (p = 0.007) and Day 2 (p = 0.033). At Day 1, NIRS monitoring with VOT showed significantly faster tissue oxygen saturation downslope (p = 0.018) and slower upslope (p = 0.04) in patients with high D4-SOFA.DiscussionIn our cohort of major traumas, sublingual microcirculation and peripheral microvascular reactivity were significantly more impaired early after trauma in those patients who developed more severe organ dysfunctions. Our data would support the hypothesis that restoration of macrocirculation can be dissociated from restoration of peripheral and tissue perfusion, and that microvascular alterations can be associated with organ failure.

Predictive Value of Sublingual Microcirculation and Peripheral Tissue Oxygen Monitoring

Predictive Value of Sublingual Microcirculation and Peripheral Tissue Oxygen Monitoring

HIV-1 vaccination by needle-free oral injection induces strong mucosal immunity and protects against SHIV challenge

The oral mucosa is an attractive site for mucosal vaccination, however the thick squamous epithelium limits antigen uptake. Here we utilize a modified needle-free injector to deliver immunizations to the sublingual and buccal (SL/B) tissue of rhesus macaques. Needle-free SL/B vaccination with modified vaccinia Ankara (MVA) and a recombinant trimeric gp120 protein generates strong vaccine-specific IgG responses in serum as well as vaginal, rectal and salivary secretions. Vaccine-induced IgG responses show a remarkable breadth against gp70-V1V2 sequences from multiple clades of HIV-1. In contrast, topical SL/B immunizations generates minimal IgG responses. Following six intrarectal pathogenic SHIV-SF162P3 challenges, needle-free but not topical immunization results in a significant delay of acquisition of infection. Delay of infection correlates with non-neutralizing antibody effector function, Env-specific CD4+ T-cell responses, and gp120 V2 loop specific antibodies. These results demonstrate needle-free MVA/gp120 oral vaccination as a practical and effective route to induce protective immunity against HIV-1.

Which impacted mandibular third molar positions or bone morphology shapes can have greater risk of accidental displacement of tooth roots into the sublingual soft tissues?

Aim: We aimed to find out which impacted mandibular third molar (3M) positions or bone morphology types at the 3M site could lead to the potential risk of accidental displacement of 3M root fragments into the sublingual soft tissues. Materials and Methods: The sample consisted of 103 patients with an impacted mandibular 3M (vertical, mesioangular, or horizontal position) seen on at least one side using cone beam computed tomography images. The bone morphology at the 3M sites were classified as round shape, lingual concavity, and lingual extended. The relationship between the roots and the sublingual soft tissues was classified as: 1) type A, in which some amount of bone appears between the root and the soft tissues; 2) type B, in which the root apex is located at the lingual cortical plate; and 3) type C, in which the root apex has penetrated into the soft tissues. Results: Binary logistic regression analysis showed that the odds of lingual extended bone morphology are 2.61 times greater for potential root displacement in combined types B and C, compared to round shape. Conclusion: These results suggest that impacted mandibular 3Ms with the lingual extended morphology type are more likely to cause root fragment displacement into the sublingual soft tissues. Keywords: Third molar; mandible; cone beam computed tomography; iatrogenic displacement; sublingual space.

Persistent Oral Infection Caused by an Iatrogenic Displacement of a Lower Third Molar Root to the Sublingual Space

ABSTRACTAim and objectiveDescribe a case of sublingual displacement of a root of the lower third molar, as well as highlight the importance of using appropriate complementary imaging tests and provide the clinician with information on how to prevent and treat this complication.BackgroundThe displacement of the lower third molar is an accidental event that can lead to serious complications, which is usually the cause of inadequate surgical technique.Case descriptionA 69-year-old man presented pain and trismus due to local infection 1 month after a tooth extraction. After a misdiagnosis of retained root and surgical exploration, a cone-beam computed tomography (CBCT) showed a displaced root into the sublingual tissues. A surgical procedure under local anesthesia was carried out to easily remove the displaced tooth root, which resulted in complete healing of soft tissues and disappearance of signs of local infection.ConclusionThe use of CBCT is essential for the identification and location of the dislocated tooth and the removal of the fragment, as it allows a three-dimensional assessment providing adequate surgical planning which resulted in complete remission of all symptoms.Clinical significanceFew reports have been published to describe this rare complication of dental extractions that support key points for proper management: (1) the use of three-dimensional imaging for proper evaluation and establishment of exact location; (2) careful surgical planning; (3) precise surgery. Successful tooth removal is the main goal of treatment, which usually results in control of associated signs and symptoms.How to cite this articleSousa NWA, de Moraes ATL, Guimarães DM,et al.Persistent Oral Infection Caused by an Iatrogenic Displacement of a Lower Third Molar Root to the Sublingual Space. J Contemp Dent 2019;9(3):155–158.

Formulation and Characterization of Fast-Dissolving Sublingual Film of Iloperidone Using Box-Behnken Design for Enhancement of Oral Bioavailability.

Iloperidone is a second-generation antipsychotic drug which is used for the treatment of schizophrenia and has very low aqueous solubility and bioavailability. This drug also undergoes first-pass metabolism. The aim of this work is to formulate fast-dissolving sublingual films of iloperidone to improve its bioavailability. Sublingual films were prepared by solvent casting method. Hydroxypropyl methyl cellulose E5, propylene glycol 400, and transcutol HP were optimized using Box-Behnken three-level statistical design on the basis of disintegration time and folding endurance of films. Iloperidone:hydroxypropyl-β-cyclodextrin kneaded complex was used in films instead of plain drug due to its low solubility. Optimized film was further evaluated for drug content, pH, dissolution studies, ex vivo permeation studies, and pharmacokinetic studies in rats. The optimized film disintegrated within 30s. The in vitro dissolution of the film showed 80.3 ± 3.4% drug dissolved within first 5min. In ex vivo permeation studies using sublingual tissue, flux achieved within first 15min by film was around 117.1 ± 0.35 (mcg/cm2/h) which was ten times more than that of plain drug. This formulation showed excellent uniformity. AUC and Cmax of film were significantly higher (p < 0.001) as compared to plain drug and relative bioavailability of the films was 148% when compared to the plain drug. Thus, this study showed optimized fast-dissolving sublingual film to improve permeation and bioavailability of iloperidone. Fast-dissolving films will be customer-friendly approach for geadiatric schizophrenic patients.

Protective Effect of Increased Zinc Supply against Oxidative Damage of Sublingual Gland in Chronic Exposure to Cadmium: Experimental Study on Rats.

Cadmium is one of the main chemical pollutants found in the daily environment of developed countries. Cigarettes are a significant source of that metal, which makes it important in terms of oral cavity health. The aim of this study was to determine if increased supply of zinc in chronic exposure to cadmium might protect the sublingual gland structure against oxidative damage. The experiment took 12 months and was conducted on 72 adult male rats. They were randomized into 9 groups. Eight groups received cadmium in drinking water (as CdCl2) at 5 or 50 mg Cd/dm3 and/or zinc (as ZnCl2) at 30 or 60 mg Zn/dm3. The control group received regular water. In the sublingual gland of all animal groups, levels of oxidative parameters were measured. The oxidative stress index was calculated as a TOS/TAS ratio. Cadmium exposure at 5 mg and 50 mg Cd/dm3 induced oxidative stress in the sublingual glands of the rats. Cadmium reduced the TAS and GSH levels and increased LPO, H2O2, TOS, and OSI. In cadmium exposure conditions, increasing the supply of zinc by 79% or 151%, as compared to the standard dietary intake of this microelement, completely prevented the reduction of TAS and GSH levels and accumulation of LPO, H2O2, and TOS in the examined gland at both exposure levels to that metal. The outcome data confirm the protective effect of increased zinc intake on the sublingual gland tissue in chronic cadmium exposure.

Surgical management of plunging ranulas: a 10-year case series in South East Queensland.

Plunging ranulas are rare mucous extravasation pseudocysts that arise in the floor of the mouth and pass into the submandibular space of the neck. The aim of this study was to investigate the diagnosis, surgical management and outcomes of patients with a plunging ranula at our institution in South East Queensland over a 10-year period. A retrospective analysis of adult patients diagnosed with and treated for plunging ranula between 2006 and 2016 at Logan Hospital was conducted. Patient demographics, preoperative investigations, surgical management and post-operative outcomes were collected from medical records. A total of 18 adult patients were treated for plunging ranula. Of the 18 cases, 17 were treated via transoral excision of the sublingual gland. The mean age at presentation was 28.8 years with a 3:1 female to male predominance. Fifty-six percent of patients were of Polynesian descent. The success rate was 94% with only one patient experiencing recurrence and requiring re-excision of remnant sublingual gland tissue. Three patients (17%) developed complications related to post-operative bleeding. There was a slight predominance for right-sided disease (56%) compared with left and one case of bilateral plunging ranulas in this series. This study demonstrates that excision of the sublingual gland is an effective and safe treatment for plunging ranula. The majority of plunging ranulas occur in patients aged <30 years with a higher incidence in patients of Polynesian heritage, which is consistent with previous studies suggesting a possible underlying genetic predisposition for this condition.

Mucosal and cutaneous capnometry for the assessment of tissue hypoperfusion.

In critically ill patients, tissue hypoperfusion is an important cause leading to multi-organ dysfunction and death, and it cannot always be detected by measuring standard global hemodynamic and oxygen-derived parameters. Gastric intramucosal partial pressure of carbon dioxide (PCO2) as measured by gastric tonometry has been recognized to be of clinical value as a prognostic factor, in assessing the effects of particular therapeutic interventions, and as an end-point of resuscitation. However, this technique has several limitations that have hampered its implementation in clinical practice. The sublingual tissue bed has been shown to be damaged in models of shock, and microcirculatory changes in this area may indicate imminent changes in other important organs. The measurement of sublingual mucosal PCO2 (PslCO2) by sublingual capnography is technically simple, noninvasive and gives near instantaneous results. Clinical studies have established that high PslCO2 values and, more especially, high PslCO2 gap (PslCO2 - arterial PCO2) values are correlated with impaired microcirculatory blood flow and a poor outcome in critically ill patients. Sublingual capnography seems to be the ideal noninvasive monitoring tool to evaluate the severity of shock states and the adequacy of tissue perfusion. However, clinical studies are needed to determine the clinical utility of PslCO2 gap monitoring as end-point target to guide resuscitation in critically ill patients.

Development of nanostructured lipid carriers for intraoral delivery of Domperidone

The oral cavity displays one potentially attractive route that is not associated with gastric transit and hepatic first pass metabolism. However, poorly soluble active candidates require a drug delivery system to facilitate their transport through oral biological membranes. To this end, nanostructured lipid carriers (NLC) were loaded with Domperidone and produced with high pressure homogenization. NLC were characterized regarding particle size, particle size distribution, zeta potential, entrapment efficiency and crystallinity. Moreover, permeability studies through the buccal mucosa were performed using in-vitro and ex-vivo models. Palmitic acid (solid lipid) and oleic acid (liquid lipid) in the ratio 9:1 were able to dissolve the highest amount of Domperidone. Particle diameters of about 280nm and monomodal size distribution were obtained. A zeta potential higher than −30mV was observed over a period of 28days indicating good physical stability. NLC dispersed in saliva did not agglomerate and were actively internalized by buccal TR 146 cells without causing adverse side effects. Ex-vivo studies confirmed that Domperidone permeated the entire tissue. This leads to the conclusion that NLC are appropriate carrier systems that facilitate the transport of poorly soluble drugs across buccal and sublingual tissue.

Sublingual targeting of STING with 3′3′-cGAMP promotes systemic and mucosal immunity against anthrax toxins

Anthrax is caused by Bacillus anthracis, a zoonotic bacterial pathogen affecting humans and livestock worldwide. The current human anthrax vaccine, anthrax vaccine adsorbed (AVA), is an injected vaccine with a cumbersome administration schedule and fails to promote mucosal immunity. Bacterial enterotoxins, which stimulate production of the cyclic nucleotide cAMP are effective experimental mucosal vaccine adjuvants, but their inherent toxicity has precluded their use in humans. We investigated whether cyclic dinucleotides that target Stimulator of Interferon Gamma Genes (STING) in mammalian cells could represent an alternative to bacterial enterotoxins as adjuvant for sublingual immunization and promotion of mucosal immunity and secretory IgA responses in addition to systemic immunity. We found that sublingual immunization of mice with Bacillus anthracis protective antigen (PA) and the STING ligand 3′3′-cGAMP promotes PA-specific serum IgG Ab responses of the same magnitude as those induced after immunization with PA and the experimental adjuvant cholera toxin (CT). Interestingly, this STING ligand also promoted serum anti-PA IgA and IgA-producing cells in the bone marrow. Furthermore, the saliva of mice immunized with the STING ligand exhibited similar levels of PA-specific IgA Abs as groups immunized with CT as adjuvant. The adjuvant activity of 3′3′-cGAMP was associated with mixed Th1, Th2, and Th17 responses. This STING ligand also induced rapid IFN-β and IL-10 responses in sublingual tissues and cervical lymph nodes, and TGF-β responses in the cervical lymph nodes, which could contribute to promoting IgA responses after sublingual immunization.

Isolation and in vitro permeation of phenylpropylamino alkaloids from Khat (Catha edulis) across oral and intestinal mucosal tissues

Ethnopharmacological relevanceKhat, the leaves of Catha edulis, is used as a “natural amphetamine-like” stimulant in eastern and southern Africa, as well as in the Arabian Peninsula. Leaves are masticated to elicit a state of euphoria. Although the psychostimulatory effects of the leaves are attributed to the presence of phenylpropylamino alkaloids (i.e. cathinone, cathine and norephedrine), the extent of permeation of these alkaloids across the oral and intestinal mucosa has not been established. Materials and methodsCathinone was isolated in the form of the oxalate salt from young buds, following acid-base extraction. High performance countercurrent chromatography (HPCCC) was used to isolate cathine and norephedrine, following borohydride reduction of a mixture of the three alkaloids. The in vitro permeability of these three alkaloids in their pure form, as well as in a crude extract, was evaluated across Caco-2 cell monolayers and across excised porcine intestinal, sublingual and buccal tissues. ResultsThe purities of the isolated cathine and norephedrine were in excess of 90%, thereby proving that HPCCC can be applied for efficient separation of these alkaloids from extracts of Khat. The apparent permeability (Papp) coefficients for the Khat alkaloids in their pure form were all above 1.0×10−6cm/s, indicating that the transport of the three alkaloids across the selected biological membranes is comparable to that of the highly permeable reference compound, caffeine. Although readily transported across the various membranes, the alkaloids were transported to a lesser extent when present in a leaf extract, suggesting that other phytochemicals present in the extract influence their permeation. ConclusionsThese results provide evidence that chewing of Khat contributes to the buccal and sublingual absorption of the psychoactive alkaloids in the bloodstream directly across the oral mucosal membranes. In addition, it confirms that these metabolites will be readily absorbed from the gastrointestinal tract when swallowed.

Multi-layered nanofibrous mucoadhesive films for buccal and sublingual administration of drug-delivery and vaccination nanoparticles - important step towards effective mucosal vaccines

Nanofibre-based mucoadhesive films were invented for oromucosal administration of nanocarriers used for delivery of drugs and vaccines. The mucoadhesive film consists of an electrospun nanofibrous reservoir layer, a mucoadhesive film layer and a protective backing layer. The mucoadhesive layer is responsible for tight adhesion of the whole system to the oral mucosa after application. The electrospun nanofibrous reservoir layer is intended to act as a reservoir for polymeric and lipid-based nanoparticles, liposomes, virosomes, virus-like particles, dendrimers and the like, plus macromolecular drugs, antigens and/or allergens. The extremely large surface area of nanofibrous reservoir layers allows high levels of nanoparticle loading. Nanoparticles can either be reversibly adsorbed to the surface of nanofibres or they can be deposited in the pores between the nanofibres. After mucosal application, nanofibrous reservoir layers are intended to promote prolonged release of nanoparticles into the submucosal tissue. Reversible adsorption of model nanoparticles as well as sufficient mucoadhesive properties were demonstrated. This novel system appears appropriate for the use in oral mucosa, especially for sublingual and buccal tissues. To prove this concept, trans-/intramucosal and lymph-node delivery of PLGA-PEG nanoparticles was demonstrated in a porcine model. This system can mainly be used for sublingual immunization and the development of “printed vaccine technology”.

増大傾向を示した舌下腺による静止性骨空洞の長期観察例

Lingual bone defects are usually asymptomatic local concavities of the lingual cortical bone of the mandible, frequently caused by soft-tissue inclusion. In most cases, the soft tissue is the submandibular gland. Only 7 cases of developing lingual bone defects were previously reported in Japan, but none of these cases involved the sublingual gland. We report a rare case of a developing lingual bone defect associated with the sublingual gland, which we followed up for 21 years. The patient was a 58-year-old man who had a developing lingual bone defect in the anterior right side of the mandible. A panoramic X-ray film showed an oval radiolucent area in the apical region of the lower right canine and first premolar tooth. Moreover, electric pulp testing confirmed that all teeth in the lower-right quadrant were vital. A panoramic X-ray film obtained at a nearby dental clinic demonstrated a radiolucent area 15 years before presentation to our department, and the area showed evidence of gradual expansion. Computed tomography revealed a local concavity in the lingual cortical bone of the right side of the mandible. The concavity measured 13 × 9 mm. The signal intensity of the inner tissue of the bone defect was nearly equivalent to that of the sublingual gland on T1-, T2-weighted magnetic resonance imaging. The suspected clinical diagnosis was a lingual bone defect. We performed an incisional biopsy to rule out a neoplastic lesion, and the histopathological diagnosis was normal sublingual gland tissue. On the basis of these results, the final diagnosis was a developing lingual bone defect associated with the sublingual gland. Two years have elapsed, but there is no swelling or induration in the right sublingual gland. The radiolucent area on the panoramic X-ray film showed decreased radiolucency and increased bone volume. Moreover, computed tomography revealed that the concavity of the bone had shrunk.

Formulation and evaluation of sublingual delivery of piroxicam using thermosensitive polymer with an inverted Franz diffusion cell.

The aim of the study was to prepare a sublingual formulation for piroxicam using a thermosensitive polymer and to evaluate its permeation through porcine sublingual mucosa. Formulation technique utilized the transition property of poloxamer from solution state at room temperature to gel state at oromucosal temperature (37 °C). The permeation of the drug was evaluated using an inverted Franz diffusion cell technique that allowed the dosage form to be directly applied onto the substrate with required volume of saliva. The formulation was characterized for microscopy of the piroxicam crystals, sol-gel transition property and in-vitro diffusion study. Poloxamer-based formulation enhanced solubility and increased permeability of the piroxicam. Poloxamer formulation with 0.1% w/w piroxicam delivered a cumulative amount of 11.99 ± 7.82 and 11.23 ± 1.79 μg/cm(2), while non-poloxamer formulation delivered 3.57 ± 2.20 and 4.60 ± 6.90 μg/cm(2) with 0.1 and 0.5 ml artificial saliva, respectively, through porcine sublingual tissue in 6 h. A similar delivery profile was observed for 0.05% w/w piroxicam formulation as well.

Evaluation of the Microstat™ sublingual PCO2 monitor in ambulatory patients.

Physicians often need to measure arterial PCO2 in clinical practice. Arterial blood gas sampling is typically available only in hospitals and may be unpleasant for patients. Minimally invasive techniques for measuring PCO2 offer the potential for overcoming these limitations. The MicroStat monitor non-invasively measures PCO2 in the sublingual tissues, which should track arterial PCO2 in hemodynamically stable patients. This was aprospective observational study. Patients undergoing routine cardiac catheterization were recruited. Following arterial cannulation, two sequential sublingual PCO2 measurements were taken and a contemporaneous arterial sample was sent for blood gas analysis. For each subject we calculated the mean sublingual-arterial CO2 gradient and the test-retest sublingual PCO2 difference. Twenty-five patients were studied. Mean sublingual-arterial PCO2 gradient was +6.8mmHg (95% limits of agreement -3.0 to 16.6mmHg). Test-retest difference was 3.4mmHg (95% limits of agreement -1.1 to 7.9mmHg), p=0.11 (Wilcoxon test), repeatability was 11mmHg. The MicroStat sublingual PCO2 monitor over-estimates arterial PCO2 with wide limits of agreement. Test-retest repeatability was poor. Use of sublingual PCO2 monitoring with the MicroStat monitor cannot currently replace blood gas sampling.

Neutrophils negatively regulate induction of mucosal IgA responses after sublingual immunization.

Induction of mucosal IgA capable of providing a first line of defense against bacterial and viral pathogens remains a major goal of needle-free vaccines given via mucosal routes. Innate immune cells are known to play a central role in induction of IgA responses by mucosal vaccines, but the relative contribution of myeloid cell subsets to these responses has not firmly been established. Using an in vivo model of sublingual vaccination with Bacillus anthracis edema toxin (EdTx) as adjuvant, we examined the role of myeloid cell subsets for mucosal secretory IgA responses. Sublingual immunization of wild-type mice resulted in a transient increase of neutrophils in sublingual tissues and cervical lymph nodes. These mice later developed Ag-specific serum IgG responses, but not serum or mucosal IgA. Interestingly, EdTx failed to increase neutrophils in sublingual tissues of IKKβΔMye mice, and these mice developed IgA responses. Partial depletion of neutrophils before immunization of wild-type mice allowed the development of both mucosal and serum IgA responses. Finally, co-culture of B cells with neutrophils from either wild-type or IKKβΔMye mice suppressed production of IgA, but not IgM or IgG. These results identify a new role for neutrophils as negative regulators of IgA responses.

Massive Lingual and Sublingual Haematoma following Postextractive Flapless Implant Placement in the Anterior Mandible

Dental implants placement in the anterior mandible with flap or flapless technique is a routine procedure and is considered to be safe. However, serious life-threatening complications may occur. We report the first case of massive lingual and sublingual haematoma following postextractive implant placement in the anterior mandible with flapless technique. A 45-year-old female patient underwent placement of four immediately postextractive implants in the anterior mandible using flapless technique. During the procedure, the patient referred intense acute pain and worsening sign of airway obstruction, dysphagia, dyspnea, and speech difficulties. Bimanual compression of the mouth floor, lingual surface of the mandible, and submental skin was maintained for approximately 25 minutes in order to stop the bleeding. Computerized tomography highlighted the massive lingual and sublingual haematoma. The symptoms and signs had almost completely resolved in the next 48 hours. The prevention of these complications is mandatory with clinical and CT analyses, in order to highlight mandibular atrophy and to select carefully the correct length and angulation of bone drilling and to keep more attention to the flapless technique considering the elevation of a lingual mucoperiosteal flap to access the mandibular contour intraoperatively and to protect the sublingual soft tissues and vasculature in high risk cases.

Establishment and first characterization of a sublingual epithelial and immune cell co-culture model

We describe here the establishment and first characterization of a co-culture model of human epithelial sublingual cells (HO-1-u-1 cell line) and human dendritic cells derived from human peripheral blood monocytes (PBMC). Cell culture conditions for HO-1-u-1 cells were optimized. First characterization of phenotypic features by electron microscopy and fluorescence imaging revealed resemblance to sublingual tissue specimen from healthy donors. Successful co-culturing of epithelial and dendritic cells (DCs) was confirmed by confocal laser scanning microscopy. Stimulation of HO-1-u1 cells alone and the epithelial/DC co-culture by incubation with liposomes, virosomes and influenza virus lead reproducibly to the release of inflammatory cytokine GM-CSF. This co-culture model may be suitable for elucidation of mechanisms involved in the immune response at the sublingual epithelium as well as for the evaluation of novel topical vaccines, potentially replacing cumbersome ex vivo and in vivo methods currently in place.