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BACKGROUND: Mesenchymal stromal cells (MSCs) isolated from the periodontal ligament (hPDL-MSCs) have a high therapeutic potential, presumably due to their immunomodulatory properties. The interaction between hPDL-MSCs and immune cells is reciprocal and executed by diverse cytokine-triggered paracrine and direct cell-to-cell contact mechanisms. For the first time, this study aimed to directly compare the contribution of various mechanisms on this reciprocal interaction using different in vitro co-culture models at different inflammatory milieus. METHODS: Three co-culture models were used: indirect with 0.4 µm-pored insert, and direct with or without insert. After five days of co-culturing mitogen-activated CD4+ T lymphocytes with untreated, interleukin (IL)-1ß, or tumor necrosis factor (TNF)-α- treated hPDL-MSCs, the CD4+ T lymphocyte proliferation, viability, and cytokine secretion were investigated. The gene expression of soluble and membrane-bound immunomediators was investigated in the co-cultured hPDL-MSCs. RESULTS: Untreated hPDL-MSCs decreased the CD4+ T lymphocyte proliferation and viability more effectively in the direct co-culture models. The direct co-culture model without inserts showed a strikingly higher CD4+ T lymphocyte cell death rate. Adding IL-1ß to the co-culture models resulted in substantial CD4+ T lymphocyte response alterations, whereas adding TNF resulted in only moderate effects. The most changes in CD4+ T lymphocyte parameters upon the addition of IL-1ß or TNF-α in a direct co-culture model without insert were qualitatively different from those observed in two other models. Additionally, the co-culture models caused variability in the immunomediator gene expression in untreated and cytokine-triggered hPDL-MSCs. CONCLUSION: These results suggest that both paracrine and cell-to-cell contact mechanisms contribute to the reciprocal interaction between hPDL-MSCs and CD4+ T lymphocytes. The inflammatory environment affects each of these mechanisms, which depends on the type of cytokines used for the activation of MSCs' immunomodulatory activities. This fact should be considered by comparing the outcomes of the different models.
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Linfócitos T CD4-Positivos , Técnicas de Cocultura , Células-Tronco Mesenquimais , Comunicação Parácrina , Ligamento Periodontal , Humanos , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/imunologia , Ligamento Periodontal/citologia , Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD4-Positivos/imunologia , Imunomodulação , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Comunicação Celular , Interleucina-1beta/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Citocinas/metabolismoRESUMO
BACKGROUND: Double-lumen tubes (DLTs) are the preferred device for lung isolation. Conventional DLTs (cDLT) need a bronchoscopic position control. Visualisation of correct DLT positioning could be facilitated by the use of a video double-lumen tube (vDLT). During the SARS-CoV-2-pandemic, avoiding aerosol-generation was suggesting using this device. In a large retrospective series, we report both general and pandemic related experiences with the device. METHODS: All anesthesia records from patients aged 18 years or older undergoing surgery from April 1st, 2020 to December 31st, 2021 in the department of thoracic surgery requiring intraoperative lung isolation were analyzed retrospectively. RESULTS: During the investigation period 343 left-sided vDLTs (77.4%) and 100 left-sided cDLTs (22.6%) were used for one lung ventilation. In the vDLT group bronchoscopy could be reduced by 85.4% related to the cDLT group. Additional bronchoscopy to reach or maintain correct position was needed in 11% of the cases. Other bronchoscopy indications occured in 3.6% of the cases. With cDLT, in 1% bronchoscopy for other indications than conforming position was observed. CONCLUSIONS: The Ambu® VivaSight™ vDLT is an efficient, easy-to-use and safe airway device for the generation of one lung ventilation in patients undergoing thoracic surgery. The vDLT implementation was achieved easily with full interchangeability to the left-sided cDLT. Using the vDLT can reduce the need for aerosol-generating bronchoscopic interventions by 85.4%. Continuous video view to the carina enabling position monitoring of the DLT without need for bronchoscopy might be beneficial for both employee's and patient's safety.
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COVID-19 , Ventilação Monopulmonar , Procedimentos Cirúrgicos Torácicos , Humanos , Estudos Retrospectivos , SARS-CoV-2 , Pandemias/prevenção & controle , Intubação Intratraqueal , Broncoscopia , Aerossóis e Gotículas RespiratóriosRESUMO
OBJECTIVES: The use of lasers for debonding adhesively luted ceramic restorations is a rather recent oral laser application in dentistry. The removal of all-ceramic restorations in the mouth can often be a troublesome task. A novel method for the debonding of ceramic restorations without damaging the restorations is Er:YAG laser irradiation. The aim of this study was to evaluate the Er:YAG laser for debonding procedures of different dental ceramics and to identify appropriate laser settings. MATERIAL AND METHODS: Lithium disilicate, zirconium-reinforced lithium silicate, feldspatic ceramic, and zirconium dioxide were investigated. Ten ceramic rectangular-shaped specimens with 1 and 2 mm thickness were produced from each material. All specimens were irradiated with four different power settings 1.5; 2.5; 3.5; 4.5 W, pulse duration 50 µs, laser repetition rate 10 Hz, time of irradiation 10 s. The transmitted energy was measured with a powermeter. Additionally the suitability of the Er:YAG laser to remove the adhesively bonded ceramic and the time until loss of retention was evaluated. RESULTS: The transmission rate for 1 and 2 mm platelets was determined for zirconium-reinforced lithium silicate at 54.6%/35.6%, lithium disilicate at 53.2%/35.7%, zirconium dioxide at 40.6%/32.4%, and for the feldspathic ceramic at 19.4%/10.1%. For zirconium-reinforced lithium silicate and zirconium dioxide 2.5 W (250 mJ/10 Hz) was an appropriate energy level for effective debonding. Whereas for lithium disilicate and for feldspathic ceramic, 4.5 W (450 mJ/10 Hz) is required for efficient debonding. CONCLUSIONS: There are differences regarding transmission rates between ceramic types for the Er:YAG laser light and additionally depending on the type of ceramic different energy settings should be used for adequate debonding. Based on our in-vitro experiments we recommend 2.5 W for zirconium-reinforced lithium silicate and zirconium dioxide and 4.5 W for lithium disilicate and feldspatic ceramic. Transmission rates of different ceramic types and varying influences of thicknesses and bonding materials should be considered to adjust the laser parameters during laser debonding of adhesively luted all-ceramic restorations.
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BACKGROUND: Airway management can be challenging in neonates and infants. The Pierre Robin sequence (PRS) is a condition characterized by micrognathia, glossoptosis and airway obstruction. The airway management of these patients poses great challenges for anesthesiologists and pediatricians alike. To date, there has been no direct comparison of the hyperangulated GlideScope® Spectrum LoPro (GLP), the straight GlideScope® Spectrum Miller (GSM), a conventional Macintosh (MC) and a conventional Miller blade (ML) in patients with PRS. METHODS: For this purpose, 90 anesthesiologists (43 with limited experience, 47 with extensive experience) performed orotracheal intubation on an Air-Sim® Pierre Robin X manikin using GLP, GSM, MC and ML in randomized order. 'Time-to-vocal-cords', 'time-to-intubate', 'time-to-ventilate', the severity of oral-soft-tissue-trauma and the subjective evaluation of each device were recorded. RESULTS: A significantly faster and better view of the vocal cords and lower oral-soft-tissue-trauma was achieved using the GLP (p<0.001). Though, there were no significant differences in the 'time-to-intubate' or 'time-to-ventilate'. The highest intubation success rate was found with GSM and the lowest with GLP (GSM 100%, ML 97.8%, MC 96.7%, GLP 93.3%). When using the videolaryngoscopes, there were no undetected esophageal intubations but in six cases prolonged attempts of intubation (>120s) with the GLP. In the sub-group with extensive experience, we found significantly shorter intubation times for the GSM and ML. The GLP was the tool of choice for most participants, while the conventional MC received the lowest rating. CONCLUSIONS: Videolaryngoscopy leads to increased safety for the prevention of undetected esophageal intubation in the airway management in a PRS manikin. Hyperangulated blades may ensure a good and fast view of the vocal cords and low oral-soft-tissue-trauma but pose a challenge during the placement of the tube. Specific skills and handling seem to be necessary to ensure a safe tube placement with this sort of blades.
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Laringoscópios , Síndrome de Pierre Robin , Lesões dos Tecidos Moles , Recém-Nascido , Humanos , Lactente , Anestesiologistas , Intubação Intratraqueal/efeitos adversos , Manuseio das Vias Aéreas , Manequins , LaringoscopiaRESUMO
The aim of this study was the evaluation of the in vitro efficacy of a carbon dioxide (CO2) laser, a tetracalcium phosphate/dicalcium phosphate anhydrate (TP/DP) desensitizer and the combination of the desensitizer and additional CO2 laser irradiation as a treatment modality for cervical dentin hypersensitivity. A total of 48 dental specimens, prepared from extracted human premolars and molars, were divided into four groups: a control group, a TP/DP desensitizer paste group, a CO2 laser (10.600-nm wavelength) group, and a paste and laser group. The specimens were coated with nail varnish except in the marked area and were then immersed in 2% methylene blue dye for 1 h. The specimens were then washed, dried, and cut longitudinally. Thereafter, photos of 40 dentin specimens were taken and evaluated. The area of penetration was assessed and reported as percentage of the dentin surface area. Additionally eight dental specimens were examined with the aid of a scanning electron microscope and evaluated. Significant differences in the penetration depth were found for all experimental groups compared to the control group. The lowest penetration area was detected in the paste-laser group (16.5%), followed by the laser (23.7%), the paste (48.5%), and the control group (86.2%). The combined treatment of the CO2 laser and a TP/DP desensitizer was efficient in sealing the dentinal surface and could be a treatment option for cervical dentin hypersensitivity.
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Sensibilidade da Dentina , Dentina , Humanos , Dentina/efeitos da radiação , Sensibilidade da Dentina/tratamento farmacológico , Sensibilidade da Dentina/radioterapia , Dióxido de Carbono/farmacologia , Microscopia Eletrônica de Varredura , LasersRESUMO
Peptide human leukocyte antigen (pHLA) targeting therapeutics like T-cell receptor based adoptive cell therapy or bispecific T cell engaging receptor molecules hold great promise for the treatment of cancer. Comprehensive pre-clinical screening of therapeutic candidates is important to ensure patient safety but is challenging because of the size of the potential off-target space. By combining stabilized peptide-receptive HLA molecules with microarray printing and screening, we have developed an ultra-high-throughput screening platform named ValidaTe that enables large scale evaluation of pHLA-binder interactions. We demonstrate its potential by measuring and analyzing over 30.000 binding curves for a high-affinity T cell Engaging Receptor towards a large pHLA library. Compared to a dataset obtained by conventional bio-layer interferometry measurements, we illustrate that a massively increased throughput (over 650 fold) is obtained by our microarray screening, paving the way for use in pre-clinical safety screening of pHLA-targeting drugs.
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Neoplasias , Peptídeos , Humanos , Peptídeos/química , Receptores de Antígenos de Linfócitos T , Biblioteca de PeptídeosRESUMO
Introduction: Human periodontal ligament-derived mesenchymal stromal cells (hPDL-MSCs) exhibit a tight bi-directional interaction with CD4+ T lymphocytes. The hPDL-MSCs' immunomodulatory abilities are drastically enhanced by pro-inflammatory cytokines via boosting the expression of various immunomediators. 25-hydroxyvitamin D3 (25(OH)D3), the major metabolite of vitamin D3 in the blood, affects both hPDL-MSCs and CD4+ T lymphocytes, but its influence on their interaction is unknown. Methods: Therefore, primary hPDL-MSCs were stimulated in vitro with tumor necrosis factor (TNF)-α a or interleukin (IL)-1ß in the absence and presence of 25(OH)D3 followed by an indirect co-culture with phytohemagglutinin-activated CD4+ T lymphocytes. The CD4+ T lymphocyte proliferation, viability, and cytokine secretion were analyzed. Additionally, the expression of various immunomediators in hPDL-MSCs was investigated, and their implication was verified by using pharmacological inhibitors. Results: 25(OH)D3 significantly counteracted the suppressive effects of IL-1ß-treated hPDL-MSCs on CD4+ T lymphocyte proliferation, whereas no effects were observed in the presence of TNF-α. Additionally, 25(OH)D3 significantly increased the percentage of viable CD4+ T lymphocytes via TNF-α- or IL-1ß-treated hPDL-MSCs. It also caused a significant decrease in interferon-γ, IL-17A, and transforming growth factor-ß productions, which were triggered by TNF-α-treated hPDL-MSCs. 25(OH)D3 significantly decreased the production of various immunomediators in hPDL-MSCs. Inhibition of two of them, prostaglandin E2 and indoleamine-2,3-dioxygenase-1, partially abolished some of the hPDL-MSCs-mediated effects of 25(OH)D3 on CD4+ T lymphocytes. Conclusion: These data indicate that 25(OH)D3 influences the immunomodulatory activities of hPDL-MSCs. This modulatory potential seems to have high plasticity depending on the local cytokine conditions and may be involved in regulating periodontal tissue inflammatory processes.
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Células-Tronco Mesenquimais , Fator de Necrose Tumoral alfa , Humanos , Fator de Necrose Tumoral alfa/metabolismo , Células Cultivadas , Ligamento Periodontal/metabolismo , Calcifediol/farmacologia , Citocinas/metabolismo , Células-Tronco Mesenquimais/metabolismoRESUMO
OBJECTIVES: This study aimed to assess levels of biomarkers associated with inflammation and tissue destruction in peri-implant crevicular fluid (PICF) of implants provided with customized or standard healing abutments during early implant healing. MATERIALS AND METHODS: Thirty implants were placed in 22 patients with partial posterior edentulism. Subsequently, test group implants (n=15) received one-piece titanium abutments that were fabricated using computer-aided design/computer-aided manufacturing (CAD/CAM). Control group implants (n=15) were provided with standard abutments. PICF collection and standardized periapical radiographs were carried out at suture removal one week later, following crown delivery after 3 months and at 6 months. Expression of C-reactive protein (CRP), interferon-γ, tumor necrosis factor (TNF)-α, interleukin (IL)-1α, IL-1ß, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12A, IL-17A, macrophage inflammatory protein (MIP)-1α, matrix metalloproteinase (MMP)-13, osteopontin, osteoactivin, Receptor Activator of NF-κB (RANK), and TGF-ß were analyzed using a multiplex ELISA kit. RESULTS: Both groups showed a significant decrease in protein expression of CRP, IL-1ß, IL-6, IL-8, MIP-1α, osteopontin, osteoactivin, and TGF-ß, while MMP-13 levels increased during the observation period. A rise in OPG and RANK levels was detected among customized abutments. Expression of CRP was higher, whereas IL-1ß, IL-1α, and MIP-1α were decreased in control compared to test group implants after 6 months. Marginal bone loss did not depend on abutment modality. CONCLUSIONS: Both abutment types showed distinctive temporal expression of inflammatory biomarkers during 6 months following implant placement. TRIAL REGISTRATION: ISRCTN98477184, registration date 18/05/2022 CLINICAL RELEVANCE: Customized healing abutments exert similar effects on inflammation during early implant healing compared to standard healing abutments.
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Implantes Dentários , Humanos , Quimiocina CCL3 , Osteopontina , Projetos Piloto , Interleucina-6 , Interleucina-8 , Fator de Necrose Tumoral alfa/análise , Inflamação , Interleucina-1alfa , Fator de Crescimento Transformador beta , Desenho Assistido por Computador , Dente Suporte , TitânioRESUMO
A 10-year-old female mixed breed dog was presented for thyroid scintigraphy due to a cervical mass. Apart from 99m-Technetium-pertechnetate (Tc-pertechnetate) scintigraphy, a second scintigraphy using 99m-Technetium sestamibi (Tc-MIBI) was performed because of additional hypothyroidism suspective for a "cold" nodule and as screening for metastases.Twenty minutes following intravenous injection of 38 MBq Tc-pertechnetate, a "hot" cervical as well as a "hot" intrathoracal nodule were seen with an uptake of 8.40 and 0.25 %, respectively. The second scintigraphy was performed 20 minutes after intravenous injection of 364 MBq Tc-MIBI and 70 minutes following the first. After subtraction of pertechnetate activity and decay correction, both nodules showed an uptake of 0.99 and 0.03 %. Additionally, both thyroid lobes were visible in the thyroid loge with a weak MIBI-uptake. For both lesions, the ratio Tc-uptake/Tc-MIBI-uptake was 8.48 and 8.33, respectively.Following the extirpation of the cervical mass, histopathology revealed atrophied healthy thyroid tissue almost completely displaced by a well-differentiated follicular thyroid carcinoma.This case report describes performance, utility and calculative correction of consecutive pertechnetate- and MIBI-scan, that enable a visualization of hypofunctional thyroid tissue.Therefore and because of their similar MIBI metabolic activity, both nodules were considered to be dystopic tissue/metastases so that this dog had to be classified as prognostically less favorable WHO stabe IV. Different from human patients, both scintigraphies should be performed shortly after another in dogs in order to avoid the necessity of a second anesthetic procedure. A reliable qualitative/visual evaluation of the MIBI-scan is therefore not possible, so that a quantitative assessment using the uptake after calculative correction of the pertechnetate activity is recommended.
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Adenocarcinoma Folicular , Doenças do Cão , Neoplasias da Glândula Tireoide , Nódulo da Glândula Tireoide , Humanos , Feminino , Cães , Animais , Pertecnetato Tc 99m de Sódio , Nódulo da Glândula Tireoide/diagnóstico por imagem , Nódulo da Glândula Tireoide/patologia , Nódulo da Glândula Tireoide/veterinária , Tecnécio , Tecnécio Tc 99m Sestamibi , Neoplasias da Glândula Tireoide/diagnóstico por imagem , Neoplasias da Glândula Tireoide/veterinária , Cintilografia , Adenocarcinoma Folicular/veterinária , Doenças do Cão/diagnóstico por imagemRESUMO
OBJECTIVES: Polymorphonuclear leucocytes (PMNs) constitute the first line of host defence and are crucial in maintaining periodontal health. Their survival and function are modulated by mesenchymal stromal cells (MSCs) from different origin. Gingival MSCs (GMSCs) play an important role in maintaining oral health and in the initial inflammatory response. The present study aimed to investigate the effects of GMSCs on PMNs apoptosis and reactive oxygen species (ROS) production. METHODS: PMNs were either directly incubated with untreated, interleukin (IL)-1ß- or tumour necrosis factor (TNF)-α-treated GMSCs or stimulated with their conditioned media. Resulting ROS production was evaluated by dichlorofluorescin diacetate staining, whereas PMNs apoptosis was assessed by Annexin V staining, followed by flow cytometry analysis. RESULTS: While conditioned media of untreated and TNF-α-treated GMSCs did not affect apoptosis of PMNs, it was significantly delayed by conditioned media of GMSCs treated with IL-1ß. In direct co-culture, GMSCs exerted anti-apoptotic effects on PMNs independently of the previous stimulation. However, the strongest impact was observed by IL-1ß-treated GMSCs. ROS production of PMNs was not influenced by GMSCs or their conditioned media. CONCLUSION: This study demonstrates for the first time the immunomodulatory properties of GMSCs towards PMNs, revealing that IL-1ß enhances anti-apoptotic effects of GMSCs.
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Células-Tronco Mesenquimais , Técnicas de Cocultura , Meios de Cultivo Condicionados/farmacologia , Gengiva , Humanos , NeutrófilosRESUMO
BACKGROUND: Barrier membranes and bone substitute are major tools of guided tissue regeneration (GTR) after periodontal disease. Integrity of the periodontal ligament plays a key role in periodontal health, but its functionality fails to be fully re-established by GTR after disease or trauma. Microtissue models suggest an in vivo-like model to develop novel GTR approaches due to its three-dimensionality. This study aims to assess the effects of collagen membranes and bone substitute on cell viability, adhesion and gene expression of regenerative and inflammatory biomarkers by periodontal ligament cell (PDLC) microtissues. METHODS: Human PDLC microtissues and monolayers were cultured on collagen membranes or bone substitute. After 24 hours incubation, metabolic activity, focal adhesion, mRNA and protein production of collagen-type-I (COL1A1), periostin (POSTN), vascular endothelial growth factor (VEGF), angiogenin (ANG), interleukin (IL)6 and IL8 were measured by resazurin-based toxicity assay, focal adhesion staining, quantitative polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. RESULTS: PDLC microtissues and monolayers were viable on collagen membranes and bone substitute, but microtissues were less metabolically active. Dominant staining of actin filaments was found in PDLC microtissues on collagen membranes. COL1A1, POSTN, VEGF, ANG and IL6 were modulated in PDLC microtissues on bone substitute, while there were no significant changes on collagen membranes. PDLC monolayers showed a different character of gene expression changes. CONCLUSIONS: PDLC microtissues and monolayers react diversely to collagen membranes and bone substitute. Further descriptive and mechanistic tests will be required to clarify the potential of PDLC microtissues as in vivo-like model for GTR.
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Substitutos Ósseos , Ligamento Periodontal , Substitutos Ósseos/farmacologia , Colágeno/farmacologia , Colágeno Tipo I , Regeneração Tecidual Guiada Periodontal , Humanos , Membranas Artificiais , Fator A de Crescimento do Endotélio VascularRESUMO
A 6-month-old French bulldog was presented due to chronic large intestinal diarrhea of 4â months duration. The diagnostic procedures initiated by the referring veterinarian had resulted in a tentative diagnosis of chronic enteropathy, however treatment consisting of elimination diet as well as antibiotic, anthelmintic, anti-inflammatory and immunosuppressive medication had been unsuccessful. By means of endoscopy and histological examination, pronounced erosions and ulcerations of the colonic mucosa were detected. Fluorescence in situ hybridization enabled the identification of invasive Escherichia coli within the colonic mucosa and colonic macrophages, allowing for the diagnosis of granulomatous colitis. The dog showed complete remission of clinical signs following 8â weeks of treatment with enrofloxacin. This case report describes the first successful treatment of granulomatous colitis with enrofloxacin in a French bulldog puppy in Germany and is intended to sensitize the reader to this disease in (young) dogs.
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Doença de Crohn , Doenças do Cão , Animais , Antibacterianos/uso terapêutico , Doença de Crohn/tratamento farmacológico , Doença de Crohn/veterinária , Diarreia/tratamento farmacológico , Diarreia/veterinária , Doenças do Cão/diagnóstico , Doenças do Cão/tratamento farmacológico , Cães , Hibridização in Situ Fluorescente/veterináriaRESUMO
OBJECTIVES: Nd:YAG and Er:YAG lasers have been previously used as an adjunct in periodontal therapy. The aim of this single-blinded randomized controlled clinical trial was to evaluate the efficacy of a combined application of Nd:YAG and Er:YAG laser irradiation in periodontal treatment. MATERIALS AND METHODS: Twenty-two patients with at least one site of ≥ 6 mm periodontal probing depth (PPD) after mechanical debridement with curettes and sonic instruments at periodontal reevaluation were included in the study. Patients were randomly allocated at a 1:1 ratio to either a combined Nd:YAG/Er:YAG laser therapy (test group) or a "turned off" laser therapy (control group). The Nd:YAG laser was used for periodontal pocket deepithelialization and to stabilize the resulting blood clot. The Er:YAG laser was primarily used for root surface modification. PPD (mm), clinical attachment level (CAL, mm), and bleeding on probing (BOP, +/-) at the site of laser treatment were evaluated at baseline and 2 months after treatment. RESULTS: The mean improvements from baseline to 2-month follow-up for PPD were significantly better in the laser group (2.05 ± 0.82 mm) compared to the control group (0.64 ± 0.90 mm; p = 0.001). Likewise, the gain in CAL was significantly better in the laser group (1.50 ± 1.10 mm) than in the control group (0.55 ± 1.01mm; p = 0.046). CONCLUSIONS: The combined application of Nd:YAG and Er:YAG laser irradiation as an adjunct to conventional non-surgical therapy showed a significant beneficial effect on periodontal treatment results. CLINICAL RELEVANCE: Combined Nd:YAG and Er:YAG laser irradiation could be a useful procedure additionally to conventional non-surgical periodontal therapy to improve periodontal treatment results. CLINICAL TRIAL REGISTRATION: ISRCTN registry #ISRCTN32132076.
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Lasers de Estado Sólido , Doenças Periodontais , Alumínio , Raspagem Dentária , Érbio , Humanos , Lasers de Estado Sólido/uso terapêutico , Neodímio , ÍtrioRESUMO
BACKGROUND: Video laryngoscopy is an effective tool in the management of difficult pediatric airway. However, evidence to guide the choice of the most appropriate video laryngoscope (VL) for airway management in pediatric patients with Pierre Robin syndrome (PRS) is insufficient. Therefore, the aim of this study was to compare the efficacy of the Glidescope® Core™ with a hyperangulated blade, the C-MAC® with a nonangulated Miller blade (C-MAC® Miller) and a conventional Miller laryngoscope when used by anesthetists with limited and extensive experience in simulated Pierre Robin sequence. METHODS: Forty-three anesthetists with limited experience and forty-three anesthetists with extensive experience participated in our randomized crossover manikin trial. Each performed endotracheal intubation with the Glidescope® Core™ with a hyperangulated blade, the C-MAC® with a Miller blade and the conventional Miller laryngoscope. "Time to intubate" was the primary endpoint. Secondary endpoints were "time to vocal cords", "time to ventilate", overall success rate, number of intubation attempts and optimization maneuvers, Cormack-Lehane score, severity of dental trauma and subjective impressions. RESULTS: Both hyperangulated and nonangulated VLs provided superior intubation conditions. The Glidescope® Core™ enabled the best glottic view, caused the least dental trauma and significantly decreased the "time to vocal cords". However, the failure rate of intubation was 14% with the Glidescope® Core™, 4.7% with the Miller laryngoscope and only 2.3% with the C-MAC® Miller when used by anesthetists with extensive previous experience. In addition, the "time to intubate", the "time to ventilate" and the number of optimization maneuvers were significantly increased using the Glidescope® Core™. In the hands of anesthetists with limited previous experience, the failure rate was 11.6% with the Glidescope® Core™ and 7% with the Miller laryngoscope. Using the C-MAC® Miller, the overall success rate increased to 100%. No differences in the "time to intubate" or "time to ventilate" were observed. CONCLUSIONS: The nonangulated C-MAC® Miller facilitated correct placement of the endotracheal tube and showed the highest overall success rate. Our results therefore suggest that the C-MAC® Miller could be beneficial and may contribute to increased safety in the airway management of infants with PRS when used by anesthetists with limited and extensive experience.
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Anestesistas , Intubação Intratraqueal/instrumentação , Laringoscópios/classificação , Laringoscopia/instrumentação , Manequins , Síndrome de Pierre Robin/cirurgia , Gravação em Vídeo/instrumentação , Adulto , Estudos Cross-Over , Feminino , Glote , Humanos , Lactente , Masculino , Pessoa de Meia-IdadeRESUMO
In addition to maintaining hemostasis, platelets have an important role in modulating innate and adaptive immune responses. A low platelet count has been found to be a negative prognostic factor for survival in humans and horses with critical illnesses, such as sepsis or systemic inflammatory response syndrome (SIRS). Decreased platelet aggregation, caused by in vivo activation, has been found in human patients with severe sepsis. In our prospective controlled study, we assessed platelet biology in blood samples from 20 equine SIRS cases and 120 healthy control horses. Platelet variables such as platelet count, large platelet count, clumps, plateletcrit, mean platelet volume, and mean platelet component concentration were analyzed by laser flow cytometry (Advia 2120) from K3EDTA blood and from citrate blood. Hirudin blood samples were analyzed by impedance aggregometry (Multiplate analyzer; Roche) for platelet aggregation, including spontaneous aggregation and aggregation by 4 different agonists: adenosine diphosphate (ADPtest), ADP + prostaglandin E1 (ADPtestHS), arachidonic acid (ASPItest), and collagen (COLtest). SIRS cases had significantly lower platelet counts in K3EDTA blood (p < 0.0001) compared to control horses. There were no significant differences in aggregation values between SIRS cases and controls. Non-surviving SIRS horses did not have statistically significant lower platelet counts or lower aggregation values for COLtest, ADPtest, or ADPtestHS compared to surviving SIRS horses, although 5 non-survivors were thrombocytopenic.
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Plaquetas/fisiologia , Doenças dos Cavalos/fisiopatologia , Agregação Plaquetária , Contagem de Plaquetas/veterinária , Síndrome de Resposta Inflamatória Sistêmica/veterinária , Animais , Estudos de Casos e Controles , Feminino , Alemanha , Doenças dos Cavalos/sangue , Cavalos , Masculino , Estudos Prospectivos , Síndrome de Resposta Inflamatória Sistêmica/sangue , Síndrome de Resposta Inflamatória Sistêmica/fisiopatologiaRESUMO
BACKGROUND: Previous in vitro studies have proven laser fluorescence measurement using a 655-nm Indium Gallium Arsenide Phosphide (InGaAsP) based diode laser radiation to be a useful tool to detect subgingival calculus. The aim of this prospective study was to evaluate the 655-nm InGaAsP diode laser in detecting subgingival calculus in patients with periodontal disease compared with photographic assessment during periodontal surgery. METHODS: Twelve patients (six women, six men) aged between 21 and 75 years with periodontitis scheduled for periodontal surgery were included in this prospective study. All laser fluorescence measurements were made before periodontal surgery. Intraoperatively a mucoperiostal flap was performed, subgingival calculus was visualized, and photographic images were taken. The presence of calculus was recorded for each evaluated site. RESULTS: A total of 115 tooth surface sites of 32 teeth from the 12 patients were evaluated before (laser) and during surgery (image). Compared with image evaluation the laser assessment showed a sensitivity of 0.70 (CI0.025 0.53 to CI0.975 0.83) and a specificity of 0.97 (CI0.025 0.85 to CI0.975 0.99). The overall probability to correctly detect subgingival calculus with the laser (accuracy) was 0.82 (CI0.025 0.74 to CI0.975 0.88). CONCLUSIONS: The 655-nm diode laser was able to detect subgingival calculus. Hence, the 655 nm diode laser may be used as an additional tool for calculus detection in non-surgical periodontal therapy.
Assuntos
Índio , Doenças Periodontais , Adulto , Idoso , Cálculos Dentários , Feminino , Humanos , Lasers Semicondutores/uso terapêutico , Pessoa de Meia-Idade , Estudos Prospectivos , Adulto JovemRESUMO
Transplanted mesenchymal stem/stromal cells (MSCs) are a promising and innovative approach in regenerative medicine. Their regenerative potential is partly based upon their immunomodulatory activities. One of the most investigated immunomediators in MSCs, such as in periodontal ligament-derived MSCs (hPDLSCs), is indoleamine-2,3-dioxygenase-1 (IDO-1) which is upregulated by inflammatory stimuli, like cytokines. However, there are no data concerning continuing IDO-1 expression in hPDLSCs after the removal of inflammatory stimuli, such as cytokines and toll-like receptor (TLR) agonist-2 and TLR-3. Hence, primary hPDLSCs were stimulated with interleukin (IL)-1ß, tumor necrosis factor (TNF)-α, interferon (IFN)-γ, TLR-2 agonist Pam3CSK4 or TLR-3 agonist Poly I/C. IDO-1 gene and protein expression and its enzymatic activity were measured up to five days after removing any stimuli. IL-1ß- and TNF-α-induced IDO-1 expression and enzymatic activity decreased in a time-dependent manner after cessation of stimulation. IFN-γ caused a long-lasting effect on IDO-1 up to five days after removing IFN-γ. Both, TLR-2 and TLR-3 agonists induced a significant increase in IDO-1 gene expression, but only TLR-3 agonist induced significantly higher IDO-1 protein expression and enzymatic activity in conditioned media (CM). IDO-1 activity of Poly I/C- and Pam3CSK4-treated hPDLSCs was higher at one day after removal of stimuli than immediately after stimulation and declined to basal levels after five days. Among all tested stimuli, only IFN-γ was able to induce long-lasting IDO-1 expression and activity in hPDLSCs. The high plasticity of IDO-1 expression and its enzymatic activity in hPDLSCs due to the variable cytokine and virulence factor milieu and the temporal-dependent responsiveness of hPDLSCs may cause a highly dynamic potential of hPDLSCs to modulate immune responses in periodontal tissues.
Assuntos
Citocinas/farmacologia , Indolamina-Pirrol 2,3,-Dioxigenase/metabolismo , Ligamento Periodontal/citologia , Células-Tronco/enzimologia , Receptores Toll-Like/agonistas , Células Cultivadas , Humanos , Indolamina-Pirrol 2,3,-Dioxigenase/genética , Lipopeptídeos/farmacologia , Poli I-C/farmacologia , Células-Tronco/efeitos dos fármacos , Células Estromais/efeitos dos fármacos , Células Estromais/enzimologia , Receptores Toll-Like/metabolismoRESUMO
Human periodontal ligament stromal cells (hPDLSCs) and gingival mesenchymal stromal cells (hGMSCs) are resident mesenchymal stromal cells (MSCs) of the periodontal tissue. The lipopolysaccharide (LPS) from Porphyromonas gingivalis is structurally distinct from that of other Gram-negative bacteria, and earlier studies linked this structural difference to a distinct virulence activity and the ability to activate toll-like receptor 2 (TLR-2), besides TLR-4 as commonly occurring upon LPS challenge. Later studies, in contrast, argue that TLR-2 activation by P. gingivalis LPS is due to lipoprotein contamination. In the present study, we aimed to define the influence of structure versus purity of P. gingivalis LPS on the immune response of hPDLSCs and hGMSCs. Cells were stimulated with commercially available "standard" P. gingivalis LPS, "ultrapure" P. gingivalis LPS, or "ultrapure" Escherichia coli LPS, and the expression of interleukin- (IL-) 8, IL-6, monocyte chemoattractant protein- (MCP-) 1, TLR-2, and TLR-4 was evaluated. The contribution of TLR-4 to the LPS-induced response was assessed using the specific TLR-4 inhibitor TAK-242. "Standard" P. gingivalis LPS induced significantly higher IL-8, IL-6, and MCP-1 production compared to the "ultrapure" LPS preparations, with no significant difference detectable for "ultrapure" LPS from P. gingivalis and E. coli. By using TAK-242, the response of hPDLSCs and hGMSCs to "ultrapure" LPS preparations was effectively inhibited to the levels comparable to those of nonstimulated controls. In contrast, high levels of response to "standard" LPS were observed, even in the presence of TAK-242. Our data show that the response of MSCs from periodontal tissue to LPS depends more on the purity of the LPS preparation than on the LPS source. Even a small amount of contaminating lipoproteins can drastically enhance the hPDLSCs' and hGMSCs; responsiveness to P. gingivalis LPS, which might also contribute to the progression of periodontal disease.
Assuntos
Lipopolissacarídeos/farmacologia , Células-Tronco Mesenquimais/citologia , Porphyromonas gingivalis/citologia , Sobrevivência Celular/fisiologia , Células Cultivadas , Quimiocina CCL2/metabolismo , Ensaio de Imunoadsorção Enzimática , Humanos , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Células-Tronco Mesenquimais/efeitos dos fármacos , Ligamento Periodontal/citologia , Ligamento Periodontal/metabolismo , Reação em Cadeia da Polimerase , Receptor 2 Toll-Like/metabolismo , Receptor 4 Toll-Like/metabolismoRESUMO
Human periodontal ligament stem cells (hPDLSCs) play an important role in periodontal tissue homeostasis and regeneration. The function of these cells in vivo depends largely on their immunomodulatory ability, which is reciprocally regulated by immune cells via cytokines, particularly interferon (IFN)-γ, tumor necrosis factor (TNF)-α, and interleukin (IL)-1ß. Different cytokines activate distinct signaling pathways and might differently affect immunomodulatory activities of hPDLSCs. This study directly compared the effect of IFN-γ, TNF-α, or IL-1ß treated primary hPDLSCs on allogenic CD4+ T lymphocyte proliferation and apoptosis in an indirect co-culture model. The effects of IFN-γ, TNF-α, and IL-1ß on the expression of specific immunomodulatory factors such as intoleamine-2,3-dioxygenase-1 (IDO-1), prostaglandin E2 (PGE2), and programmed cell death 1 ligand 1 (PD-L1) and ligand 2 (PD-L2) in hPDLSCs were compared. The contribution of different immunomodulatory mediators to the immunomodulatory effects of hPDLSCs in the indirect co-culture experiments was assessed using specific inhibitors. Proliferation of CD4+ T lymphocytes was inhibited by hPDLSCs, and this effect was strongly enhanced by IFN-γ and IL-1ß but not by TNF-α. Apoptosis of CD4+ T lymphocytes was decreased by hPDLSCs per se. This effect was counteracted by IFN-γ or IL-1ß. Additionally, IFN-γ, TNF-α, and IL-1ß differently regulated all investigated immunomediators in hPDLSCs. Pharmacological inhibition of immunomediators showed that their contribution in regulating CD4+ T lymphocytes depends on the cytokine milieu. Our data indicate that inflammatory cytokines activate specific immunomodulatory mechanisms in hPDLSCs and the expression of particular immunomodulatory factors, which underlies a complex reciprocal interaction between hPDLSCs and CD4+ T lymphocytes.
Assuntos
Citocinas/metabolismo , Imunomodulação , Células-Tronco Mesenquimais/imunologia , Ligamento Periodontal/citologia , Adolescente , Adulto , Apoptose/efeitos dos fármacos , Antígeno B7-H1/metabolismo , Biomarcadores/metabolismo , Linfócitos T CD4-Positivos/imunologia , Proliferação de Células/efeitos dos fármacos , Ciclo-Oxigenase 2/metabolismo , Inibidores de Ciclo-Oxigenase/farmacologia , Dinoprostona/metabolismo , Hematopoese/efeitos dos fármacos , Humanos , Imunomodulação/efeitos dos fármacos , Indolamina-Pirrol 2,3,-Dioxigenase/metabolismo , Inflamação/patologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Adulto JovemRESUMO
BACKGROUND: In human medicine, extremely high CRP (C-reactive protein) concentrations > 100 mg/l are indicators of bacterial infection and the need of antibiotic treatment. Similar decision limits for septic pneumonia are recommended for dogs but have not yet been evaluated for other organ systems. The aim of the retrospective study was to investigate the prevalence and evaluate dogs with CRP concentrations > 100 mg/l regarding the underlying etiology, the affected organ system and the prognostic significance. RESULTS: Prevalence of CRP > 100 mg/l was investigated in dogs presented between 2014 and 2015 and was 12%. For evaluation of etiology and organ systems, dogs with CRP > 100 mg/l presented between 2014 and 2016 were enrolled. Dogs were classified into 4 main disease categories, i.e. inflammatory, neoplastic, tissue damage or "diverse". Diseases were assigned to the affected organ system. If an organ classification was not possible, dogs were classified as "multiple". 147 dogs with CRP 101-368 mg/l were included and classified into disease categories: 86/147 (59%) with inflammatory etiology (among these, 23/86 non-infectious, 44/86 infectious (33/44 bacterial), 19/86 inflammation non-classifiable), 31/147 (21%) tissue damage, 17/147 (12%) neoplastic (all malignant) and 13/147 (9%) diverse diseases. The affected organ systems included 57/147 (39%) multiple, 30/147 (20%) trauma, 21/147 (14%) gastrointestinal tract, 10/147 (7%) musculoskeletal system, 8/147 (5%) respiratory tract, 7/147 (5%) urinary/reproductive tract, 6/147 (4%) skin/subcutis/ear, 6/147 (4%) central/peripheral nervous system and 2/147 (1%) heart. The disease group (p = 0.081) or organ system (p = 0.17) did not have an impact on CRP. Based on CRP, a detection of bacterial infection was not possible. The prognostic significance was investigated by determining the 3-months survival and hospitalization rate in a subgroup with known outcome. The 3-months survival rate was 46/73 (63%) while the majority 66/73 (90%) of patients was hospitalized. CONCLUSIONS: CRP concentrations > 100 mg/l are occasionally seen in a clinic population. They indicate a severe systemic disease of various etiologies with guarded prognosis. Extremely high CRP concentrations do not allow a conclusion of the underlying etiology or an identification of bacterial inflammation.