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The aim of this anatomical study was to investigate the genial spinal canal histologically and to reconstruct it three-dimensionally to improve understanding of its anatomy and to reveal any differences between dentate and edentulous specimens. Two tissue blocks from the mandible between the left and right second incisors, one dentate and one edentulous, were fixed in 4.5% formaldehyde, decalcified and embedded in paraffin. Serial histological sections were prepared, stained with Azan and examined microscopically. Additionally, three-dimensional models of the blocks were reconstructed using microphotographs of the sections. The genial spinal canal in the dentate specimen contained a neurovascular bundle, which branched into a nerve innervating the incisor and a neurovascular bundle, whereas that in the edentulous specimen contained some nerves for vestibular gingival innervation and a vascular bundle. The results suggest differences in the genial spinal canal between dentate and edentulous mandibles. Further research is needed to confirm this finding. Clin. Anat., 33:1102-1108, 2020. © 2019 Wiley Periodicals, Inc.
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Mandíbula/anatomia & histologia , Mandíbula/diagnóstico por imagem , Idoso de 80 Anos ou mais , Feminino , Humanos , Imageamento Tridimensional , MasculinoRESUMO
BACKGROUND: Bacterial biofilms adhere to all tissues and surfaces in the oral cavity. Oral biofilms are responsible for the decay of human dental structures and the inflammatory degeneration of the alveolar bone. Moreover, oral biofilms on artificial materials influence the lifespan of dental prostheses and restoratives. METHODS: To investigate in vivo oral biofilm formation and growth, five different dental restorative materials were analyzed and compared to human enamel. The roughness of the materials and the human enamel control probe were measured at the start of the study. The dental restorative materials and the human enamel control probe were placed in dental splints and worn for 3 h, 24 h and 72 h. RESULTS: Scanning electron microscopy (SEM) revealed major differences between oral biofilm formation and growth on the materials compared to those on human enamel. Microbiological analyses showed that bacterial strains differed between the materials. Significant differences were observed in the roughness of the dental materials. CONCLUSIONS: It can be concluded that material roughness affects biofilm formation on dental surfaces and restoratives, but other factors, such as surface charge, surface energy and material composition, may also have an influence.
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Aderência Bacteriana/fisiologia , Biofilmes , Implantes Dentários/microbiologia , Materiais Dentários , Boca/microbiologia , Esmalte Dentário/microbiologia , Placa Dentária/microbiologia , Humanos , Propriedades de SuperfícieRESUMO
OBJECTIVES: The marginal quality of ceramic inlays was evaluated after the use of three different instrumental finishing methods in mesio-occluso-distal (mod) cavity boxes in vitro after hydrothermal loading (HTL). MATERIALS AND METHODS: Caries-free human molars were divided into three groups. Mod-cavities were conventionally prepared. Box finishing was performed in every group with rotating (RI), sonic (SI), or ultrasonic (USI) instruments. Surface roughness was examined. Twelve mod-cavities remained untreated. Continuous margin quality was evaluated with scanning electron microscopy (SEM). Ceramic inlays were cemented into cavities. After HTL microleakage, marginal and absolute marginal gaps were examined. All data were analyzed statistically. RESULTS: Significant differences were found, between cavity surface roughness of RI and SI groups, the RI and USI groups, but not between microleakage, marginal, absolute marginal gaps after HTL and in proximal marginal quality. No correlations between microleakage and marginal gaps nor between microleakage and surface roughness were found. CONCLUSION: Mod-cavity proximal box finishing with SI or USI resulted in a higher surface roughness than the use of RI. The type of the finishing method did not influence the marginal quality of ceramic inlays. For the mod-cavity finishing, the use of SI and USI could be an alternative instrumental method to conventional RI methods with a lower risk of iatrogenic damage of the adjacent teeth. CLINICAL RELEVANCE: This study allows the practitioner to better determine the proper indications and limitations of the sonic and ultrasonic instruments for mod-cavity proximal box finishing.
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Cerâmica/química , Preparo da Cavidade Dentária/instrumentação , Restaurações Intracoronárias , Ultrassom/instrumentação , Infiltração Dentária , Adaptação Marginal Dentária , Humanos , Técnicas In Vitro , Teste de Materiais , Microscopia Eletrônica de Varredura , Dente Molar , Propriedades de SuperfícieRESUMO
Facial folds and creases are established descriptive anatomical terms for structures of which the morphological characteristics and origins are not clearly defined. The aim of this study was to perform a morphological investigation of the nasolabial fold (NLF), mandibular fold (MF), deep transverse forehead (DTFC), infraorbital fold (IOF) and upper eyelid fold (UEF), correlating their phenotypes to differences in the superficial musculoaponeurotic system (SMAS), noting morphological differences and similarities. Full-graft tissue blocks of skin, subcutaneous tissue, and mimic muscles collected postmortem were studied histologically. Serial histological sections were stained with Azan. Location- and composition-specific morphological differences were determined. Histological serial section digitalization and three-dimensional reconstruction of the tissue blocks were performed. Three different types of SMAS architecture were identified. Type I SMAS consisted of parallel-aligned fibrous septa connecting the mimic muscles to the skin that covered the cheek, infraorbital and supraorbital, and forehead areas. Type II SMAS morphology appeared as a condensed Type I SMAS architecture with stronger fibrous septa and smaller fatty tissue compartments covering the lower and upper lip areas. Type III SMAS consisted of loose connective tissue covering the lower and upper eyelid regions. NLF, MF, IOF, and UEF are habitual primary folds induced by morphological changes in the underlying SMAS architecture. The secondary, accidental creases (DTFC) are cutaneous depressions derived from interacting dermal-skeletal-muscular changes without SMAS structure changes. The upper and lower eyelid wrinkles were tertiary, age-related undulating skin redundancy formations. Clin. Anat. 32:573-584, 2019. © 2019 The Authors. Clinical Anatomy published by Wiley Periodicals, Inc. on behalf of American Association of Clinical Anatomists.
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Envelhecimento da Pele/patologia , Sistema Musculoaponeurótico Superficial/anatomia & histologia , Idoso , Face/patologia , Feminino , Humanos , MasculinoRESUMO
OBJECTIVES: Bioactive glasses which degrade in aqueous solutions may release bioactive ions such as fluoride (F-) and support fluoride bioavailability in saliva. We investigated how these effects would be apparent in an in vivo experimental trial after toothbrushing in comparison with sodium fluoride and amine fluoride. MATERIAL AND METHODS: In this single-center, randomized, parallel in vivo trial with a three strata block design, where healthy subjects were randomly assigned into three groups. Each group brushed their teeth either with fluoridated bioactive glass containing dentifrice, with a sodium fluoride (NaF) containing dentifrice or with amine fluoride (AmF) containing toothpaste. Saliva was collected time intervals before, immediately after, 30, 60 and 120 min after toothbrushing. Fluoride concentration was determined in supernatant saliva and salivary sediment using a fluoride ion selective electrode. The data were evaluated statistically using non-parametric tests. RESULTS: The increase of bioactive fluoride in supernatant saliva was higher after application of NaF or AmF compared to fluoridated bioactive glass. In salivary sediment bioavailability of fluoride lasted longer after application of fluoridated bioactive glass. CONCLUSIONS: Toothbrushing with the fluoride containing bioactive glass dentifrices had positive effects on the fluoride bioavailability within two hours. Fluoride containing bioactive glass represent a new area for investigation in caries prophylaxis. The bioactive potential impact on the tooth remineralization should be examined further. TRIAL REGISTRATION: DRKS00016038 .
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Cariostáticos/análise , Cárie Dentária/prevenção & controle , Dentifrícios/química , Fluoretos/análise , Saliva/química , Fluoreto de Sódio/química , Escovação Dentária , Aminas , Disponibilidade Biológica , Humanos , Avaliação de Processos e Resultados em Cuidados de Saúde , Saliva/metabolismo , Fatores de TempoRESUMO
OBJECTIVE: Soft tissue (ST) dehiscence with graft exposure is a frequent complication of vertical augmentation. Flap dehiscence is caused by failure to achieve tension-free primary wound closure and by the impairment of flap microcirculation due to surgical trauma. Soft tissue expansion (STE) increases ST quality and quantity prior to reconstructive surgery. We hypothesized that flap preconditioning using STE would reduce the incidence of ST complications after bone augmentation and that optimized ST healing would improve the outcome of bone regeneration. MATERIALS AND METHODS: Self-filling tissue expanders were implanted in mandibular bone defects in ten beagle dogs. After expansion, alloplastic scaffolds were placed for vertical bone augmentation in STE sites and in control sites without STE pre-treatment. ST flap microcirculation was analysed using laser Doppler flowmetry. The incidence of graft exposures was evaluated after 2 weeks. Bone formation was assessed after 2 months, using histomorphometry and immunohistochemistry. RESULTS: Test sites showed significantly less impairment of perfusion and faster recovery of microcirculation after bone augmentation. Furthermore, no flap dehiscences occurred in STE sites. Bone regeneration was found in both groups; however, significantly greater formation of new bone was detected in test sites with preceding STE. CONCLUSIONS: Preconditioning using STE improved ST healing and bone formation after vertical augmentation. The combination of STE and the subsequent placement of alloplastic scaffolds may facilitate the reconstruction of severe bone defects.
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Aumento do Rebordo Alveolar/métodos , Regeneração Óssea , Expansão de Tecido , Animais , Cães , Masculino , Modelos Animais , Distribuição Aleatória , Dispositivos para Expansão de TecidosRESUMO
The biological phenomenon of cell fusion plays an important role in several physiological processes, like fertilization, placentation, or wound healing/tissue regeneration, as well as pathophysiological processes, such as cancer. Despite this fact, considerably less is still known about the factors and conditions that will induce the merging of two plasma membranes. Inflammation and proliferation has been suggested as a positive trigger for cell fusion, but it remains unclear, which of the factor(s) of the inflamed microenvironment are being involved. To clarify this we developed a reliable assay to quantify the in vitro fusion frequency of cells using a fluorescence double reporter vector (pFDR) containing a LoxP-flanked HcRed/DsRed expression cassette followed by an EGFP expression cassette. Because cell fusion has been implicated in cancer progression four human breast cancer cell lines were stably transfected with a pFDR vector and were co-cultured with the stably Cre-expressing human breast epithelial cell line. Cell fusion is associated with a Cre-mediated recombination resulting in induction of EGFP expression in hybrid cells, which can be quantified by flow cytometry. By testing a panel of different cytokines, chemokines, growth factors and other compounds, including exosomes, under normoxic and hypoxic conditions our data indicate that the proinflammatory cytokine TNF-α together with hypoxia is a strong inducer of cell fusion in human MDA-MB-435 and MDA-MB-231 breast cancer cells.
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Neoplasias da Mama/fisiopatologia , Fusão Celular/métodos , Glândulas Mamárias Humanas/fisiologia , Western Blotting , Hipóxia Celular/fisiologia , Linhagem Celular Tumoral , Feminino , Citometria de Fluxo , Vetores Genéticos/genética , Vetores Genéticos/metabolismo , Proteínas de Fluorescência Verde/metabolismo , Humanos , Integrases , Microscopia Eletrônica de Varredura , Fator de Necrose Tumoral alfa/metabolismoRESUMO
INTRODUCTION: The superficial musculoaponeurotic system (SMAS) is a well described facial functional unit in humans. SMAS connects mimic musculature to the skin having many implication in facial mimic expression. One of the various morphological and physiological analogies in human and Macaca mulatta species is the facial mimic. The present study analyzed Macaca mulatta species SMAS morphology and its facial topographical differences and compared this with human SMAS tissue morphology. MATERIAL AND METHODS: Macaca mulatta full-graft tissue blocks of skin, subcutaneous tissue and mimic muscles from five topographical different facial regions (Regio Temporalis, Regio Buccalis, Regio Infraorbitalis, Regio Angulus Oris and Regio Mandibularis) were collected postmortem from eight individuals (n = 8) at the German Primate Center, Leibniz Institute for Primate Research in Göttingen (DPZ) and studied histologically. Haematoxylin-eosin and azan stained histological serial sections of full-graft tissue blocks were analyzed and SMAS topographical differences evaluated. RESULTS: SMAS typical tissue morphology was recognized in all Macaca mulatta histological serial sections (n = 780). Regio Infraorbitalis Macaca mulatta SMAS (MmSMAS) morphology was similar to human infraorbital SMAS morphology (type I SMAS). Suborbicularis oculi fat pad was recognized in Macaca mulatta samples. Human type I similar SMAS morphology was demonstrated over Macaca mulatta Regio Temporalis and Regio Buccalis. Regio Angulus Oris and the cranial area of the Regio Mandibularis presented human type II similar SMAS morphology. Type IV MmSMAS was closely related to the parotid gland tissue presence. The cervical area of the Regio Mandibularis presented human type V similar SMAS morphology. CONCLUSIONS: SMAS is a complex fibro-musculo-adipose tissue network and probably an important pivot in Macaca mulatta facial system supporting mimic expression. This study provided insights into MmSMAS typology and similarity with human SMAS tissue morphology.
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Sistema Musculoaponeurótico Superficial , Animais , Humanos , Sistema Musculoaponeurótico Superficial/anatomia & histologia , Macaca mulatta , Face/anatomia & histologia , Bochecha/anatomia & histologia , Tela Subcutânea/anatomia & histologiaRESUMO
BACKGROUND: Different fluoride formulations may have different effects on caries prevention. It was the aim of this clinical study to assess the fluoride content, provided by NaF compared to amine fluoride, in saliva and plaque. METHODS: Eight trained volunteers brushed their teeth in the morning for 3 minutes with either NaF or amine fluoride, and saliva and 3-day-plaque-regrowth was collected at 5 time intervals during 6 hours after tooth brushing. The amount of collected saliva and plaque was measured, and the fluoride content was analysed using a fluoride sensitive electrode. All subjects repeated all study cycles 5 times, and 3 cycles per subject underwent statistical analysis using the Wilcoxon-Mann-Whitney test. RESULTS: Immediately after brushing the fluoride concentration in saliva increased rapidly and dropped to the baseline level after 360 minutes. No difference was found between NaF and amine fluoride. All plaque fluoride levels were elevated after 30 minutes until 120 minutes after tooth brushing, and decreasing after 360 minutes to baseline. According to the highly individual profile of fluoride in saliva and plaque, both levels of bioavailability correlated for the first 30 minutes, and the fluoride content of saliva and plaque was back to baseline after 6 hours. CONCLUSIONS: Fluoride levels in saliva and plaque are interindividually highly variable. However, no significant difference in bioavailability between NaF and amine fluoride, in saliva, or in plaque was found.
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Cariostáticos/farmacocinética , Placa Dentária/metabolismo , Dentifrícios/farmacocinética , Fluoretos/farmacocinética , Saliva/metabolismo , Adulto , Idoso , Aminas/farmacocinética , Disponibilidade Biológica , Estudos Cross-Over , Feminino , Humanos , Eletrodos Seletivos de Íons , Masculino , Pessoa de Meia-Idade , Fluoreto de Sódio/farmacocinética , Estatísticas não Paramétricas , Fluoretos de Estanho/farmacocinética , Escovação Dentária , Adulto JovemRESUMO
Applying antibacterial coatings to dental implant materials seems reasonable but can have negative influences on desired cell adhesion and healing. In this study, zirconia abutment specimens interacting with gingival tissue were used. The aim was to compare the influence of machined or coated zirconia surfaces on the adhesion and proliferation of human gingival fibroblasts (HGF-1). Surface modifications were performed using atmospheric plasma coating with hydroxyapatite, zinc, and copper. Zirconia specimens were divided into four groups: hydroxyapatite, hydroxyapatite with zinc oxide (ZnO), hydroxyapatite with copper (Cu), and an untreated machined surface. After the characterization of the surface conditions, the morphology of adhered HGF-1 was determined by fluorescence staining and subjected to statistical evaluation. The visual analysis of cell morphology by SEM showed flat, polygonal, and largely adherent fibroblast cells in the untreated group, while round to partially flat cells were recorded in the groups with hydroxyapatite, hydroxyapatite + ZnO, and hydroxyapatite + Cu. The cell membranes in the hydroxyapatite + ZnO and hydroxyapatite + Cu groups appeared porous. The results show that HGF-1 adhere and proliferate well on machined zirconia, while plasma coating with hydroxyapatite or hydroxyapatite mixtures does not lead to increased adhesion or proliferation.
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BACKGROUND: Recently it has been shown that radiation induces migration of glioma cells and facilitates a further spread of tumor cells locally and systemically. The aim of this study was to evaluate whether radiotherapy induces migration in head and neck squamous cell carcinoma (HNSCC). A further aim was to investigate the effects of blocking the epidermal growth factor receptor (EGFR) and its downstream pathways (Raf/MEK/ERK, PI3K/Akt) on tumor cell migration in vitro. METHODS: Migration of tumor cells was assessed via a wound healing assay and proliferation by a MTT colorimeritric assay using 3 HNSCC cell lines (BHY, CAL-27, HN). The cells were treated with increasing doses of irradiation (2 Gy, 5 Gy, 8 Gy) in the presence or absence of EGF, EGFR-antagonist (AG1478) or inhibitors of the downstream pathways PI3K (LY294002), mTOR (rapamycin) and MEK1 (PD98059). Biochemical activation of EGFR and the downstream markers Akt and ERK were examined by Western blot analysis. RESULTS: In absence of stimulation or inhibition, increasing doses of irradiation induced a dose-dependent enhancement of migrating cells (p < 0.05 for the 3 HNSCC cell lines) and a decrease of cell proliferation (p < 0.05 for the 3 HNSCC cell lines). The inhibition of EGFR or the downstream pathways reduced cell migration significantly (almost all p < 0.05 for the 3 HNSCC cell lines). Stimulation of HNSCC cells with EGF caused a significant increase in migration (p < 0.05 for the 3 HNSCC cell lines). After irradiation alone a pronounced activation of EGFR was observed by Western blot analysis. CONCLUSION: Our results demonstrate that the EGFR is involved in radiation induced migration of HNSCC cells. Therefore EGFR or the downstream pathways might be a target for the treatment of HNSCC to improve the efficacy of radiotherapy.
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Carcinoma de Células Escamosas/patologia , Carcinoma de Células Escamosas/radioterapia , Movimento Celular/efeitos dos fármacos , Movimento Celular/efeitos da radiação , Receptores ErbB/antagonistas & inibidores , Neoplasias de Cabeça e Pescoço/patologia , Neoplasias de Cabeça e Pescoço/radioterapia , Carcinoma de Células Escamosas/tratamento farmacológico , Carcinoma de Células Escamosas/enzimologia , Linhagem Celular Tumoral , Relação Dose-Resposta à Radiação , Fator de Crescimento Epidérmico/metabolismo , Receptores ErbB/metabolismo , Flavonoides/farmacologia , Raios gama , Neoplasias de Cabeça e Pescoço/tratamento farmacológico , Neoplasias de Cabeça e Pescoço/enzimologia , Humanos , MAP Quinase Quinase 1/metabolismo , Sistema de Sinalização das MAP Quinases , Fosfatidilinositol 3-Quinases/metabolismo , Quinazolinas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Sirolimo/farmacologia , Carcinoma de Células Escamosas de Cabeça e Pescoço , Serina-Treonina Quinases TOR/metabolismo , Tirfostinas/farmacologiaRESUMO
OBJECTIVE: The aim of this study was to evaluate changes in bone conduction thresholds before, during and after total stapedectomy. STUDY DESIGN: Prospective clinical study. METHODS: In 27 ears of 26 patients undergoing stapedectomy under local anesthesia, bone conduction was measured before surgery, during surgery under open oval window conditions, and after the insertion of a steel wire connective tissue prosthesis. Statistical data analysis was performed on the audiometric results. RESULTS: Under open oval window conditions, bone conduction hearing was found to be improved between 500 and 2000 Hz, but not at 4000 Hz. After insertion of the prosthesis, an additional improvement was evident at 500 and 1000 Hz, but a loss was seen at 2000 and 4000 Hz. CONCLUSION: This is the first investigation reported in which audiometry was performed under open oval window conditions during stapes surgery. Our results demonstrate that at least part of the preoperative bone conduction hearing loss in otosclerosis must be of mechanical, but not of sensorineural origin, as already suspected by Carhart. The fixed footplate suppresses cochlear micromechanics mainly at frequencies between 500 to 2000 Hz. Furthermore, the loss in bone conduction hearing at 2000 and 4000 Hz after insertion of the prosthesis indicates that rather than the surgical procedure of total removal of the footplate, other factors such as the handling of the prosthesis or its mechanical properties after insertion cause high-frequency hearing loss after stapes surgery.
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Condução Óssea/fisiologia , Otosclerose/fisiopatologia , Janela do Vestíbulo/fisiopatologia , Cirurgia do Estribo , Estribo/fisiopatologia , Adulto , Idoso , Análise de Variância , Audiometria , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prótese Ossicular , Otosclerose/cirurgia , Janela do Vestíbulo/cirurgia , Estudos Prospectivos , Resultado do TratamentoRESUMO
The authors wish to make the following correction to this paper [...].
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High performance polymers like PEEK (polyetheretherketone) and FRC (fiberreinforced composite) could substitute metallic alloys for removable partial dentures. However, these polymers require aesthetic veneering. This study was to determine the bond strength to direct composite. Specimens made of PEEK and FRC were produced and air-abraded (50 µm aluminum-oxide). Specimens were allocated to four experimental groups: Luxatemp Glaze & Bond, Scotchbond Universal, SR Nexco Connect and iBond Universal. Specimens were divided into three subgroups for short-term, long-term and no artificial aging and shear bond strength (SBS) was evaluated. SBS of specimens made of PEEK with no artificial aging showed values between 10.79-14.00 MPa, short-term artificial aging resulted in values between 3.78-13.85 MPa and after long-term artificial aging SBS decreased to 0-8.75 MPa. SBS measurement of FRC specimens resulted in values between 9.83-12.1 MPa without aging, after short-term artificial aging values decreased to 8.36-11.98 MPa and after long-term aging SBS showed a degradation to 4.52-7.82 MPa.
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Resinas Compostas , Colagem Dentária , Análise do Estresse Dentário , Estética Dentária , Teste de Materiais , Polímeros , Resistência ao Cisalhamento , Propriedades de SuperfícieRESUMO
Chipping of veneering is the most common clinical complication for zirconia restorations. Veneering composite could be a promising alternative to renew restorations. Zirconia discs (3-YSZ) were prepared with varying surface treatments and bonded to indirect composite as follows: air abrasion and Scotchbond Universal (A/SU); air abrasion and Clearfil Ceramic Primer (A/C); air abrasion and MKZ Primer (A/M); air abrasion and Monobond Plus (A/MP); silica-coating and Scotchbond Universal (S/SU); air abrasion (AP/SU), additional cold atmospheric plasma treatment, and Scotchbond Universal. An indirect composite material was then applied to the zirconia specimens. Specimens were divided into subgroups for short-term (14 days storage at 37 °C and 5000 thermal cycles) and long-term (250 days storage and 37,500 thermal cycles) artificial aging. Shear bond strength measurement (SBS) was performed, and data were analyzed by Kruskal-Wallis-test and multiple comparison testing with Dunn's correction (p ≤ 0.05). The median SBS values (MPa) of short- and long-term artificial aging were: 3.09/1.36 (A/SU); 0.77/1.43 (S/SU); 2.82/2.15 (AP/SU); 1.97/1.80 (A/C); 2.01/1.58 (A/M); and 1.70/1.68 (A/MP). For short-term artificial aging A/SU showed the highest median SBS values, whereas in the long-term trial, AP/SU showed the highest values and the difference was significant. A prolonged artificial aging decreased SBS in all groups, except S/SU. In summary, treatment with CAP can improve SBS in the long-term.
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INTRODUCTION: The superficial musculoaponeurotic system (SMAS) is a controversial functional fibro-adipose layer that connects the mimic muscles to the skin and is involved in a variety of facial mimic expressions. The presence of muscle fibers within SMAS fibrous septa is hypothetical. The present study analyzed SMAS fibrous septa composition for the existence of striated muscle cells. METHODS: Histological serial sections of the sample borders (n=107) of 19 in sano-resected and diagnosed cutaneous tumors of the midfacial region were investigated. Immunohistochemical (actin and myosin) and hematoxylin and eosin staining were performed to detect striated muscle cells in SMAS fibrous septa. RESULTS: A fibro-neuro-musculo-vascular functional unit within SMAS fibrous septa was demonstrated. SMAS striated muscle cells were morphologically independent from preparotideal and periorbital mimic muscles. Intraseptal blood vessels draining the superficial and deep SMAS vascular system were described. CONCLUSIONS: Striated muscle cells were demonstrated within SMAS fibrous septa. Nerve cells and vascular tissue together with the SMAS fibro-muscular meshwork demonstrated an autonomous operating functional unit that hypothetical modulated individual mimic expression contributing to the diversity of mimic expression. The SMAS develops with mimic muscle contractions as a synergetic effect during facial crease and fold formation processes.
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Músculo Estriado , Sistema Musculoaponeurótico Superficial , Tecido Adiposo , Face , Músculos Faciais , Neoplasias Faciais , Humanos , Imuno-HistoquímicaRESUMO
OBJECTIVES: Research involving the nose reveals important information regarding the morphology and physiology of the epithelium and its molecular response to agents. The role of nasal epithelial cells and other cell subsets within the nasal epithelium play an interesting translational split between experimental and clinical research studying respiratory disorders or pathogen reactions. With an additional technical manuscript including a detailed description of important technical aspects, tips, tricks, and nuances for a successful culturing of primary, human nasal epithelial cells (NAEPCs), we here aim to improve the process of communication between experimentalists and physicians, supporting the purpose of a fruitful work for future translational projects. METHODS: Based on previous work on various complex culture models of subject-derived NAEPCs, this additional manuscript harmonizes previously published facts combined with own experiences for a trouble-free implementation in laboratories. RESULTS: A well-designed experimental question is essential prior to the establishment of different NAEPCs culture models. The correct method of cell extraction from the nasal cavity is essential and represent an important basis for successful culture work. Prior enzymatic processing of biopsy specimens, cell culture materials, collagenization procedure, culture conditions, and choice of culture medium are some important practical notes that increase the quality of the culture. Moreover, protocols on imaging techniques including histologic and electron microscopy must be adapted for NAEPC culture. Adapted flow cytometric protocols and transepithelial electrical resistance measurements can add valuable information. OUTLOOK: A successful culturing of NAEPCs can provide an important basis for genetic studies and the implementation of omics-science, which is increasingly receiving broad attention in the scientific community. The common aim of in vitro 'mini-noses' will be a breakthrough in laboratories aiming to perform research under in vivo conditions. Here, organoid models are interesting models presenting a basis for translational studies.
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Células Epiteliais , Cavidade Nasal , Técnicas de Cultura de Células , Epitélio , Humanos , Mucosa NasalRESUMO
Understanding the response to viral infection in the context of respiratory diseases is of significant importance. Recently, there has been more focus on the role of the nasal epithelium in disease modeling. Here, we provide an overview of different submerged, organotypic 3D and spheroid cell culture models of nasal epithelial cells, which were used to study asthma and allergy with a special focus on virus infection. In detail, this review summarizes the importance, benefits, and disadvantages of patient-derived cell culture models of nasal- and bronchial epithelial cells, including a comparison of these cell culture models and a discussion on why investigators should consider using nasal epithelial cells in their research. Exposure experiments, simple virus transduction analyses as well as genetic studies can be performed in these models, which may provide first insights into the complexity of molecular signatures and may open new doors for drug discovery and biomarker research.
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Asma/virologia , Células Epiteliais/virologia , Mucosa Nasal/virologia , Animais , Técnicas de Cultura de Células/métodos , Humanos , Mucosa Respiratória/virologia , Viroses/virologiaRESUMO
Although the nose, as a gateway for organism-environment interactions, may have a key role in asthmatic exacerbation, the rhinobiome of exacerbated children with asthma was widely neglected to date. The aim of this study is to understand the microbiome, the microbial immunology, and the proteome of exacerbated children and adolescents with wheeze and asthma. Considering that a certain proportion of wheezers may show a progression to asthma, the comparison of both groups provides important information regarding clinical and phenotype stratification. Thus, deep nasopharyngeal swab specimens, nasal epithelial spheroid (NAEsp) cultures, and blood samples of acute exacerbated wheezers (WH), asthmatics (AB), and healthy controls (HC) were used for culture (n = 146), 16 S-rRNA gene amplicon sequencing (n = 64), and proteomic and cytokine analyses. Interestingly, Proteobacteria were over-represented in WH, whereas Firmicutes and Bacteroidetes were associated with AB. In contrast, Actinobacteria commonly colonized HCs. Moreover, Staphylococcaceae, Enterobacteriaceae, Burkholderiaceae, Xanthobacteraceae, and Sphingomonadaceae were significantly more abundant in AB compared to WH and HC. The α-diversity analyses demonstrated an increase of bacterial abundance levels in atopic AB and a decrease in WH samples. Microbiome profiles of atopic WH differed significantly from atopic AB, whereby atopic samples of WH were more homogeneous than those of non-atopic subjects. The NAEsp bacterial exposure experiments provided a disrupted epithelial cell integrity, a cytokine release, and cohort-specific proteomic differences especially for Moraxella catarrhalis cultures. This comprehensive dataset contributes to a deeper insight into the poorly understood plasticity of the nasal microbiota, and, in particular, may enforce our understanding in the pathogenesis of asthma exacerbation in childhood.