Polymeric Nanocapsules Loaded with Lidocaine: A Promising Formulation for Topical Dental Anesthesia.

Lidocaine is the most commonly used local anesthetic worldwide, known for its rapid onset and moderate duration of anesthesia. However, it is short-lived and does not effectively promote effective topical anesthesia in the oral cavity when used alone. Our aim was to investigate whether an approximate 50% encapsulation of lidocaine in poly(ε-caprolactone) nanocapsules (LDC-Nano) would be able to increase its permeation and analgesic efficacy and reduce cytotoxicity. In this study, we characterized LDC-Nano and conducted MTT tests with HaCaT cells to assess their in vitro cytotoxicity. Additionally, in vitro permeation assays across the pig esophageal epithelium and the anesthetic efficacy of the hind paw incision model in rats were performed. Plain lidocaine (LDC) was compared with LDC-Nano and lidocaine hydrochloride plus epinephrine (LDC-Epi). The physicochemical characteristics of LDC-Nano were satisfactory (pH: 8.1 ± 0.21; polydispersity index: 0.08 ± 0.01; mean diameter (nm): 557.8 ± 22.7; and encapsulation efficiency (%): 51.8 ± 1.87) and remained stable for up to 4 months. LDC-Nano presented similar in vitro cytotoxicity to LDC but was higher than LDC-Epi (LD50: LDC = 0.48%; LDC-Nano = 0.47%; and LDC-Epi = 0.58%; p < 0.0001). Encapsulation increased the permeability coefficient about 6.6 times and about 7.5 the steady-state flux of lidocaine across the mucosal epithelium. Both encapsulation and epinephrine improved anesthesia duration, with epinephrine demonstrating superior efficacy (100% of animals were anesthetized up to 100, 30, and 20 min when LDC-Epi, LDC-nano, and LDC were used, respectively). Although LDC-Epi demonstrated superior in vivo anesthetic efficacy, the in vitro permeation and cytotoxicity of LDC-Nano indicate promising avenues for future research, particularly in exploring its potential application as a topical anesthetic in the oral cavity.

Antinociceptive effect of ultra-low dose naltrexone in a pre-clinical model of postoperative orofacial pain.

Painmanagement after oral surgeries is essential to enhance recovery, reduce negative outcomes and improve the experience of the patient. Naltrexone (NTX) is a non-selective opioid receptor antagonist that has been shown to modulate neuro-inflammation when employed in low to ultra-low doses. In addition, ultra-low dose naltrexone (ULDN) has been shown to potentiate opioids' analgesia and to have opioid-sparing effects. Herein it was investigated the effect of ULDN in a model of postoperative orofacial pain in rats, and it was tested the hypothesis that blockade of TLR4-signalling pathway contributes to its antinociceptive effect. Systemic NTX reduced heat hyperalgesia in female rats and heat and mechanical hyperalgesia in male rats after incision surgery. Combined treatment with NTX and morphine, both at ineffective doses, resulted in a significant reduction of heat hyperalgesia in male rats. NTX injection at the incision site failed to change heat hyperalgesia, but injection at the trigeminal ganglion (TG) or subnucleus caudalis (Sp5C) caused a significant reduction in heat hyperalgesia. At these sites, blockade of TLR4 impeded NTX effect. Lipopolysaccharide (LPS) injection in the intraoral mucosa resulted in facial heat hyperalgesia an increase in IL-1ß levels in the TG, which were reduced by systemic NTX. Stimulation of macrophages with LPS resulted in increase of nitric oxide, IL-1ß and CXCL-2 levels which were reduced by NTX. Altogether, these results provide evidence for an antinociceptive effect of ULDN in postoperative orofacial pain and suggest that blockade of TLR4 and downstream signaling pathway contribute to its effect.

The prognostic value of peripheral ischemic microvascular reserve in sepsis is not related to calcitonin gene-related peptide or substance P. / O valor prognóstico da reserva microvascular isquêmica periférica na sepse não está vinculado ao peptídeo relacionado ao gene da calcitonina ou à substância P.

OBJECTIVE: To evaluate the mechanisms attributed to the prognostic value of peripheral ischemic microvascular reserve in patients with sepsis. METHODS: This observational cohort study enrolled 46 consecutive septic patients in the intensive care unit between November 2020 and October 2021. After fluid resuscitation, the peripheral ischemic microvascular reserve was evaluated using the association of postocclusion reactive hyperemia with the peripheral perfusion index. Additionally, peripheral venous blood samples were used to evaluate the neuropeptide calcitonin gene-related peptide and substance P levels in the upper limb before and immediately after postocclusion reactive hyperemia. RESULTS: There was no statistically significant correlation (p > 0.05) between basal values (pg/mL) or variations from neuropeptide levels (%) and the peripheral ischemic microvascular reserve (%). CONCLUSION: Although calcitonin gene-related peptide and substance P may have a prognostic role in sepsis, these neuropeptides do not appear to contribute to peripheral ischemic microvascular reserve.

OBJETIVO: Avaliar possíveis mecanismos atribuídos ao valor prognóstico da reserva microvascular isquêmica periférica em pacientes com sepse. MÉTODOS: Este estudo de coorte observacional incluiu 46 pacientes consecutivos com sepse em uma unidade de terapia intensiva entre novembro de 2020 e outubro de 2021. Após a ressuscitação volêmica com fluidos, avaliou-se a reserva microvascular isquêmica periférica mediante a associação dos testes hiperemia reativa pós-oclusão e índice de perfusão periférica. Adicionalmente, amostras de sangue venoso periférico foram coletadas para avaliar os níveis dos neuropeptídeos substância P e peptídeo relacionado ao gene da calcitonina no membro superior do paciente antes e imediatamente após o teste de hiperemia reativa pós-oclusão. RESULTADOS: Não houve correlação estatisticamente significativa (p > 0,05) entre os valores basais ou variações dos níveis de neuropeptídeos e a reserva microvascular isquêmica periférica. CONCLUSÃO: Embora o peptídeo relacionado ao gene da calcitonina e a substância P possam desempenhar papel prognóstico na sepse, esses neuropeptídeos não parecem contribuir para a reserva microvascular isquêmica periférica.

Antinociceptive effects of bupivacaine and its sulfobutylether-ß-cyclodextrin inclusion complex in orofacial pain.

Bupivacaine hydrochloride (BVC) represents an option to produce long-lasting analgesia, and complexation in cyclodextrins has shown improvements in biopharmaceutical properties. This study aimed to characterize and test the cytotoxicity and antinociceptive effects of BVC complexed in sulfobutylether-ß-cyclodextrin (SBEßCD). The kinetics and stoichiometry of complexation and BVC-SBEßCD association constant were evaluated by phase solubility study and Job's plot. Evidence of the BVC-SBEßCD complex formation was obtained from scanning electron microscopy (SEM), infrared spectroscopy (FTIR), and differential scanning calorimetry (DSC). The cytotoxicity was evaluated in keratinocyte (HaCaT) and neuroblastoma (SH-SY5Y). Antinociceptive effects were registered via orofacial pain models: the formalin test, carrageenan-induced hyperalgesia, and postoperative pain (intraoral incision). The complex formation occurred at a 1:1 BVC-SBEßCD molar ratio, with a low association constant (13.2 M-1). SEM, DSC, and FTIR results demonstrated the host-guest interaction. The IC50% values determined in SH-SY5Y were 216 µM and 149 µM for BVC and BVC-SBEßCD, respectively (p < 0.05). There was no difference in HaCaT IC50%. In orofacial pain model, BVC-SBEßCD significantly prolonged antinociceptive effect, in about 2 h, compared to plain BVC. SBEßCD can be used as a drug delivery system for bupivacaine, whereas the complex showed long-lasting analgesic effects.

O valor prognóstico da reserva microvascular isquêmica periférica na sepse não está vinculado ao peptídeo relacionado ao gene da calcitonina ou à substância P / The prognostic value of peripheral ischemic microvascular reserve in sepsis is not related to calcitonin gene-related peptide or substance P

RESUMO Objetivo: Avaliar possíveis mecanismos atribuídos ao valor prognóstico da reserva microvascular isquêmica periférica em pacientes com sepse. Métodos: Este estudo de coorte observacional incluiu 46 pacientes consecutivos com sepse em uma unidade de terapia intensiva entre novembro de 2020 e outubro de 2021. Após a ressuscitação volêmica com fluidos, avaliou-se a reserva microvascular isquêmica periférica mediante a associação dos testes hiperemia reativa pós-oclusão e índice de perfusão periférica. Adicionalmente, amostras de sangue venoso periférico foram coletadas para avaliar os níveis dos neuropeptídeos substância P e peptídeo relacionado ao gene da calcitonina no membro superior do paciente antes e imediatamente após o teste de hiperemia reativa pós-oclusão. Resultados: Não houve correlação estatisticamente significativa (p > 0,05) entre os valores basais ou variações dos níveis de neuropeptídeos e a reserva microvascular isquêmica periférica. Conclusão: Embora o peptídeo relacionado ao gene da calcitonina e a substância P possam desempenhar papel prognóstico na sepse, esses neuropeptídeos não parecem contribuir para a reserva microvascular isquêmica periférica.

ABSTRACT Objective: To evaluate the mechanisms attributed to the prognostic value of peripheral ischemic microvascular reserve in patients with sepsis. Methods: This observational cohort study enrolled 46 consecutive septic patients in the intensive care unit between November 2020 and October 2021. After fluid resuscitation, the peripheral ischemic microvascular reserve was evaluated using the association of postocclusion reactive hyperemia with the peripheral perfusion index. Additionally, peripheral venous blood samples were used to evaluate the neuropeptide calcitonin gene-related peptide and substance P levels in the upper limb before and immediately after postocclusion reactive hyperemia Results: There was no statistically significant correlation (p > 0.05) between basal values (pg/mL) or variations from neuropeptide levels (%) and the peripheral ischemic microvascular reserve (%). Conclusion: Although calcitonin gene-related peptide and substance P may have a prognostic role in sepsis, these neuropeptides do not appear to contribute to peripheral ischemic microvascular reserve.

Mucoadhesive, Thermoreversible Hydrogel, Containing Tetracaine-Loaded Nanostructured Lipid Carriers for Topical, Intranasal Needle-Free Anesthesia.

Recent advances have been reported for needle-free local anesthesia in maxillary teeth by administering a nasal spray of tetracaine (TTC) and oxymetazoline, without causing pain, fear, and stress. This work aimed to assess whether a TTC-loaded hybrid system could reduce cytotoxicity, promote sustained permeation, and increase the anesthetic efficacy of TTC for safe, effective, painless, and prolonged analgesia of the maxillary teeth in dental procedures. The hybrid system based on TTC (4%) encapsulated in nanostructured lipid carriers (NLC) and incorporated into a thermoreversible hydrogel of poloxamer 407 (TTCNLC-HG4%) displayed desirable rheological, mechanical, and mucoadhesive properties for topical application in the nasal cavity. Compared to control formulations, the use of TTCNLC-HG4% slowed in vitro permeation of the anesthetic across the nasal mucosa, maintained cytotoxicity against neuroblastoma cells, and provided a three-fold increase in analgesia duration, as observed using the tail-flick test in mice. The results obtained here open up perspectives for future clinical evaluation of the thermoreversible hybrid hydrogel, which contains TTC-loaded NLC, with the aim of creating an effective, topical, intranasal, needle-free anesthesia for use in dentistry.

Promising potential of articaine-loaded poly(epsilon-caprolactone) nanocapules for intraoral topical anesthesia.

To determine whether the permeation capacity and analgesic efficacy of articaine (ATC) could be increased and cytotoxicity decreased by encapsulation in poly(ɛ-caprolactone) nanocapsules (ATCnano), aiming at local or topical anesthesia in dentistry. Cellular viability was evaluated (using the MTT test and fluorescence microscopy) after 1 h and 24 h exposure of HaCaT cells to ATC, ATCnano, ATC with epinephrine (ATCepi), and ATC in nanocapsules with epinephrine (ATCnanoepi). The profiles of permeation of 2% ATC and 2% ATCnano across swine esophageal epithelium were determined using Franz-type vertical diffusion cells. Analgesic efficacy was evaluated with a von Frey anesthesiometer in a postoperative pain model in rats, comparing the 2% ATC, 2% ATCnano, 2% ATCepi, and 2% ATCnanoepi formulations to 4% ATCepi (a commercially available formulation). We show that use of the nanocapsules decreased the toxicity of articaine (P<0.0001) and increased its flux (P = 0.0007). The 2% ATCepi and 4% ATCepi formulations provided higher analgesia success and duration (P<0.05), compared to 2% ATC, 2% ATCnano, and 2% ATCnanoepi. Articaine-loaded poly(ɛ-caprolactone) nanocapsules constitute a promising formulation for intraoral topical anesthesia (prior to local anesthetic injection), although it is not effective when injected in inflamed tissues for pain control, such as irreversible pulpitis.

Effects of 2-Hydroxypropil-Β-Cyclodextrin-Lidocaine on Tumor Growth and Inflammatory Response.

BACKGROUND: Antiproliferative and cytotoxic effects of lidocaine have been reported in tumor cells. However, the use of these drugs is restricted due to their short action with rapid dispersion from the injected site. The complexation of local anesthetics in 2-hydroxypropyl-ß-cyclodextrin (HP-ß-CD) is able to improve pharmacological features. OBJECTIVE: This study evaluated the antitumor effects of lidocaine and the complex HP-ß-CD-lidocaine (HP-ß-CD-lido). METHODS: In vitro;, human adenocarcinoma (HeLa) and keratinocytes (HaCaT) were exposed to lidocaine formulations and cell viability, proliferation and apoptosis induction were measured. In vivo;, Walker 256 carcinoma cells were subcutaneously injected into the plantar region of the rat right hind paw. The animals were treated with a local application of 5% lidocaine or 5% HP-ß-CD-lido. Doxorubicin (3 mg/Kg/day, intraperitoneal) was used as a positive control. Edema sizes were measured daily and the release of cytokines (TNF-α, IL-1α and CXCL-1) and prostaglandin E2 was evaluated. Histological analysis was also performed. RESULTS: HaCaT IG50 values were 846 µM and 2253 µM for lido and HP-ß-CD-lido, respectively. In HeLa cells, the IG50 was 1765 µM for lido and 2044 µM for HP-ß-CD-lido. Lidocaine formulations significantly reduced the paw edema on day 6 after Walker 256 cells inoculation. However, there were no differences in the release of inflammatory mediators in comparison to the control group. CONCLUSION: Lidocaine formulations were able to reduce the edema in vivo;, without affecting the tumor- induced inflammatory response. The antiproliferative effects of lidocaine formulations may have contributed to tumor reduction.

Physicochemical characterization and cytotoxicity of articaine-2-hydroxypropyl-ß-cyclodextrin inclusion complex.

Articaine (ATC) is one of the most widely used local anesthetics in dentistry. Despite its safety, local toxicity has been reported. This study aimed to develop an ATC-2- hydroxypropyl-ß-cyclodextrin inclusion complex (ATC HPßCD) and to assess its toxicity in vitro. The inclusion complex was performed by solubilization, followed by a fluorimetric and job plot assay to determine the complex stoichiometry. Scanning electron microscopy, DOSY- 1 H-NMR, differential scanning calorimetry (DSC), and sustained release kinetics were used to confirm the inclusion complex formation. In vitro cytotoxicity was analyzed by MTT assay and immunofluorescence in HGF cells. Fluorimetric and job plot assay determined the inclusion complex stoichiometry (ATC:HPßCD = 1:1) and complex formation time (400 min), as indicated by a strong host/guest interaction (Ka = 117.8 M - 1), complexed fraction (f = 41.4%), and different ATC and ATC HPßCD melting points (172 °C e 235 °C, respectively). The mean of cell viability was 31.87% and 63.17% for 20-mM ATC and 20-mM ATC HPßCD, respectively. Moreover, remarkable cell toxicity was observed with free ATC by immunofluorescence. These results indicate the ATC HPßCD complex could be used to improve the safety of ATC. Further research are needed to establish the anesthetic safety and effectiveness in vivo .

Comparison of antinociceptive effects of plain lidocaine versus lidocaine complexed with hydroxypropyl-ß-cyclodextrin in animal models of acute and persistent orofacial pain.

Herein, it was investigated whether a complex of lidocaine with 2-hydroxypropyl-ß-cyclodextrin (HP-ß-CD) would present a better antinociceptive profile in vivo when compared with plain lidocaine in models of orofacial pain. Plain lidocaine (LDC) and complexed lidocaine (LDC:HP-ß-CD) were initially evaluated in vitro to determine the release rate of the two formulations. Subsequently, the effect of both formulations was evaluated in independent groups of rats submitted to the orofacial formalin test, induction of facial heat hyperalgesia by capsaicin and carrageenan, and induction of facial heat and mechanical hyperalgesia by constriction of the infraorbital nerve. LDC:HP-ß-CD led to a reduction in the lidocaine release assessed in the in vitro release assay compared to plain LDC. Both formulations presented an antinociceptive effect in all models, but LDC:HP-ß-CD showed a better effect in the second phase of the formalin response, in carrageenan-induced heat hyperalgesia, and in the heat hyperalgesia associated to infraorbital nerve constriction. Our results show that complexation improved in vivo antinociceptive effects of LDC, but further studies are necessary to elucidate what properties contribute to the better effect of the complexed formulation on this models and/or what characteristics of the pain model facilitate the action of the complexed formulation.

Fungal-Host Interaction: Curcumin Modulates Proteolytic Enzyme Activity of Candida albicans and Inflammatory Host Response In Vitro.

Current treatments for Candida albicans infection are limited due to the limited number of antifungal drugs available and the increase in antifungal resistance. Curcumin is used as a spice, food preservative, flavoring, and coloring agent that has been shown to have many pharmacological activities. Thus, this study evaluated the modulatory effects of curcumin on major virulence factors associated with the pathogenicity of C. albicans. The minimum inhibitory concentration (MIC) of curcumin against C. albicans (SC5314) was determined. Biofilm formation was quantified and the proteinase and phospholipase secretion was measured. The cytotoxicity was tested in oral fibroblast cells. A cocultured model was used to analyze the gene expression of proinflammatory cytokines (IL-1ß, IL-1α, and IL-6) from host cells, as well SAP-1 and PLB-1 by RT-PCR. The MIC was between 6.25 and 12.5 µM, and the activity of proteinase enzyme was significantly decreased in biofilms treated with curcumin. However, proteinase gene expression was not downregulated after curcumin treatment. Furthermore, gene expressions of host inflammatory response, IL-1ß and IL-1α, were significantly downregulated after exposure to curcumin. In conclusion, curcumin exhibited antifungal activity against C. albicans and modulated the proteolytic enzyme activities without downregulating the gene expression. In host inflammatory response, curcumin downregulated IL-1ß and IL-1α gene expression.

Role of peripheral and central TRPV1 receptors in facial heat hyperalgesia in streptozotocin-induced diabetic rats.

There is increasing evidence that diabetes may be related to sensory changes in the trigeminal system. Long lasting facial heat hyperalgesia has been described in diabetic rats, but the mechanisms remain to be elucidated. Herein, the contribution of peripheral and central TRPV1 receptors to facial heat hyperalgesia in diabeticrats was investigated. Diabetes was induced in male Wistar rats by streptozotocin (60mg/kg, i.p) and facial heat hyperalgesia was assessed once a week up to four weeks. The role of TRPV1 receptors in the heat hyperalgesia in diabetic rats was evaluated through: 1) the ablation of TRPV1 receptors by resiniferatoxin (RTX) treatment and 2) injection of the TRPV1 antagonist, capsazepine, into the upper lip, trigeminal ganglion or medullary subarachnoid space, at doses that completed prevented the heat hyperalgesia induced by capsaicin in naïve rats. Western blot was used to estimate the changes in TRPV1 expression in diabetic rats. Diabetic rats exhibited facial heat hyperalgesia from the first up to the fourth week after streptozotocin injection, which was prevented by insulin treatment. Ablation of TRPV1-expressing fibers prevented facial hyperalgesia in diabetic rats. Capsazepine injection in all sites resulted in significant reduction of facial heat hyperalgesia in diabetic rats. Diabetic rats exhibited a significant decrease in TRPV1 expression in the trigeminal nerve, increased expression in the trigeminal ganglion and no changes in subnucleus caudalis when compared to normoglycemic ones. In conclusion, our results suggest that facial heat hyperalgesia in diabetic rats is maintained by peripheral and central TRPV1 receptors activation.

The effect of two drug delivery systems in ropivacaine cytotoxicity and cytokine release by human keratinocytes and fibroblasts.

OBJECTIVES: Modified drug delivery systems have been developed to improve pharmacological properties of local anaesthetics. However, the inflammatory potential of these formulations was not investigated. This study compared the in-vitro effects of ropivacaine (ropi) in plain, liposomal (MLV) or 2-hydroxypropyl-ß-cyclodextrin (HP-ß-CD) formulations on cell viability, apoptosis and cytokine (IL-1α, TNF-α, IL-6 and IL-10) release. METHODS: Human immortalized keratinocytes (HaCaT) and human immortalized gingival fibroblasts (HGF) were exposed to 1-100 µm ropi concentrations. The cell viability was measured by XTT and LIVE/DEAD assay. Apoptosis was performed by flow cytometry, and cytokine release was measured by ELISA assay. KEY FINDINGS: Human immortalized keratinocyte viability was reduced by ropi and both drug delivery systems. However, none of the formulations induced apoptosis. Results showed a differential regulation of IL-1α TNF-α, IL-6 and IL-10 by HaCaT and HGF. Ropi-HP-ß-CD increased twofold the IL-6 release by HGF in comparison with the control, while 100 µm ropi-MLV led to an increased release of all pro-inflammatory cytokines by HGF. CONCLUSION: The loss in cell viability was not related to cellular apoptosis. Ropi complexed with HP-ß-CD showed a similar cytokine release pattern when compared to the plain formulation. Thus, the HP-ß-CD form was a better drug carrier than the MLV form for ropivacaine drug delivery.

Comparison of liposomal and 2-hydroxypropyl-ß-cyclodextrin-lidocaine on cell viability and inflammatory response in human keratinocytes and gingival fibroblasts.

OBJECTIVES: The aim of this study was to observe the effect multilamellar liposomes (MLV) and 2-hydroxypropyl-ß-cyclodextrin (HP-ß-CD) in the in-vitro effects of lidocaine in cell viability, pro-inflammatory cytokines and prostaglandin E2 release of both human keratinocytes (HaCaT) and gingival fibroblasts (HGF) cells. METHODS: HaCaT and HGF cells were exposed to lidocaine 100-1 µm in plain, MLV and HP-ß-CD formulations for 6 h or 24 h. The formulation effects in cell viability were measured by XTT assay and by fluorescent labelling. Cytokines (IL-8, IL-6 and TNF-α) and PGE2 release were quantified by ELISA. KEY FINDINGS: MLV and HP-ß-CD formulations did not affect the HaCaT viability, which was significantly decreased by plain lidocaine after 24 h of exposure. Both drug carriers increased all cytokines released by HGF after 24-h exposure, and none of the carriers was able to reduce the PGE2 release induced by lidocaine. CONCLUSION: The effect of drug carrier in the lidocaine effects was dependent on the cell type, concentration and time of exposure. MLV and HP-ß-CD showed benefits in improving cell viability; however, both of them showed a tendency to increase cytokine release when compared to the plain solution.

Correlation between facial types and muscle TMD in women: an anthropometric approach.

Temporomandibular disorders (TMD) affecting the articular disc and/or the facial muscles are common among the population, recording a higher incidence in women age 20-40 years. The aim of this study was to investigate the correlation between facial types and muscle TMD in women. This study comprised 56 women age 18 to 49 years, seeking treatment for TMD at the School of Medicine, Federal University of São Paulo. All of the study individuals were diagnosed with muscle TMD, based on the Research Diagnostic Criteria (RDC). Facial type was determined using the Facial Brugsch Index and classified as euryprosopic (short and/or broad), mesoprosopic (average width) and leptoprosopic (long and/or narrow). The data were submitted to the Chi-square test and ANOVA-Tukey's test to conduct the statistical analysis. The faces of 27 individuals were classified as euryprosopic (48%), 18 as mesoprosopic (32%), and 11 as leptoprosopic (20%). A statistically significant difference (Chi-square, p = 0.032) was found among the facial types, in that leptoprosopic facial types showed the lowest values for muscle TMD. A greater number (p = 0.0007) of cases of muscle TMD were observed in the 20 to 39 year-old subjects than in the subjects of other age segments. In conclusion, women with euryprosopic facial types could be more susceptible to muscle TMD. Further studies are needed to investigate this hypothesis.

Correlation between facial types and muscle TMD in women: an anthropometric approach

Temporomandibular disorders (TMD) affecting the articular disc and/or the facial muscles are common among the population, recording a higher incidence in women age 20-40 years. The aim of this study was to investigate the correlation between facial types and muscle TMD in women. This study comprised 56 women age 18 to 49 years, seeking treatment for TMD at the School of Medicine, Federal University of São Paulo. All of the study individuals were diagnosed with muscle TMD, based on the Research Diagnostic Criteria (RDC). Facial type was determined using the Facial Brugsch Index and classified as euryprosopic (short and/or broad), mesoprosopic (average width) and leptoprosopic (long and/or narrow). The data were submitted to the Chi-square test and ANOVA-Tukey’s test to conduct the statistical analysis. The faces of 27 individuals were classified as euryprosopic (48%), 18 as mesoprosopic (32%), and 11 as leptoprosopic (20%). A statistically significant difference (Chi-square, p = 0.032) was found among the facial types, in that leptoprosopic facial types showed the lowest values for muscle TMD. A greater number (p = 0.0007) of cases of muscle TMD were observed in the 20 to 39 year-old subjects than in the subjects of other age segments. In conclusion, women with euryprosopic facial types could be more susceptible to muscle TMD. Further studies are needed to investigate this hypothesis.

Time-related increase of staphylococci, Enterobacteriaceae and yeasts in the oral cavities of comatose patients.

BACKGROUND/PURPOSE: The composition of oral microbiota in comatose patients remains uncertain. Some pulmonary pathogens may be found in dental biofilms or as part of the saliva microbiota. It is supposed that some pneumopathogenic microorganisms may overgrow in the mouths of comatose patients and spread to their lungs. METHODS: The oral colonization dynamics of staphylococci, Enterobacteriaceae and yeasts in nine comatose patients (group 1), and in 12 conscious patients that brushed their teeth at least twice a day (group 2) was evaluated. Both groups were followed up for 7 days after hospitalization. Daily samples of saliva were obtained, dispersed and plated on selective culture media and colony forming units of each microbial group were obtained. RESULTS: For patients in group 1, the counts of total viable bacteria, staphylococci, Enterobacteriaceae and yeasts progressively increased in a time-dependant manner. For the conscious patients of group 2, there was no increase. CONCLUSION: It would appear that concomitant consciousness and brushing teeth are determinants in controlling the selected pneumopathogen counts in resting saliva. The increase in microbial counts in comatose patients is understandable because these microorganisms could spread to the lungs.

Molecular fingerprinting methods for studies involving oral Candida albicans: [review] / Métodos de caracterização molecular para estudos envolvendo Candida albicans de origem bucal: [revisão]

OBJECTIVE: This review was aimed to discuss the literature concerning the fingerprint methods for epidemiological studies of oral-borne Candida albicans. DISCUSSION: Interest in obtaining a better understanding of the pathogenesis, epidemiology, genetics and evolution of Candida albicans has led to the development of innumerable investigations. These studies have employed fingerprinting systems, such as multilocus enzyme electrophoresis, electrophoretic karyotyping, randomly amplified polymorphic DNA, and restriction length fragment polymorphism, with and without hybridization. The efficacy of these systems has been examined at different levels of discrimination. A validation strategy has been delineated which compares two or more unrelated methods. Moreover, the different fingerprinting patterns produced could be registered in database programs and submitted to comparison with parameters of the host and characteristics of the pathogen. These procedures permit urrent and retrospective comparison of a selection of clinical and epidemiologically important strains, which could show one or several characteristics of the host or pathogen. Additionally, the sum of this growing amount of information could contribute even more to the understanding of the dynamics of infectious organisms in human populations, the complex relationship between commensalism and infection, and genetic and evolutionary mechanisms. CONCLUSIONS: Multiple molecular systems are available for studies involving C. albicans. This growing amount of information contributes to the understanding of the dynamics of this fungus in human populations.

OBJETIVO: Esta revisão discute as informações existentes acerca dos métodos de caracterização para estudos epidemiológicos envolvendo Candida albicans de origem bucal. DISCUSSÃO: O interesse no melhor entendimento da patogênese, epidemiologia, genética e evolução de C. albicans tem levado os pesquisadores à condução de inúmeras investigações. Esses estudos empregam sistemas de caracterização molecular como eletroforese de enzimas multilocus, cariotipagem por eletroforese, amplificação do DNA polimórfico ao acaso e polimorfismo dos fragmentos de restrição com e sem hidridização. A eficácia desses sistemas tem sido avaliada nos seus diferentes níveis de discriminação. Uma estratégia de validação foi delineada, a qual compara dois ou mais métodos não relacionados. Ainda, os diferentes padrões de caracterização molecular produzem dados que podem ser avaliados por programas computacionais e permite a comparação comparâmetros do hospedeiro e características do patógeno. Tais procedimentos permitem comparações correntes e retrospectivas de cepas clínicas e epidemiologicamente importantes, que podem mostrar uma ou mais características do hospedeiro ou do patógeno. A somatória do montante de informação pode contribuir para o entendimento da dinâmica dos organismos infecciosos em populações humanas, as relações complexas entre comensalismo e infecção, e mecanismos genéticos e evolutivos. CONCLUSÕES: Vários sistemas de caracterização molecular estão disponíveis para estudos envolvendo C. albicans. Este aumento de informação contribui na compreensão da dinâmica deste fungo em populações humanas.