Indirect evaluation of pit and fissure sealants : a 3D-based method validation

The aim of the present study was to compare indirect methods to assess the clinical performance of pit and fissure sealants and validate the use of 3D scanners. Sample consisted of 58 plaster models of upper and lower first permanent molars, sealed with resin sealants, as well as photographs obtained during the 18-month follow-up. Pre-established criteria were applied to categorize the sealant presence/absence and marginal integrity. Two calibrated examiners performed the evaluations, independently, using Scanning Electron Microscopy (SEM; gold-standard), Photography, 3D (CEREC In Lab) and Stereomicroscope analysis. The intra-examiner Spearman correlation was 94% e 97%, respectively, and the inter-examiner was 96%. Data was submitted to Kappa test, Spearman correlation and Receiver Operating Characteristic Curve (ROC). 3D and SEM presented good concordance; Stereomicroscope showed regular concordance with SEM and 3D (p0.05). SEM had a significant positive correlation with 3D and Stereomicroscope (r=0.76 and 0.71, respectively; p<0.01). There was significant positive correlation (r=0.65) between 3D and Stereomicroscope (p<0.01). The ROC estimated curve areas for Stereomicroscope and 3D were 0.90 (IC:0.81-0.99) and 1.0 (IC:1.0-1.0), respectively (p<0.001). Photography presented lower sensitivity and specificity (area=0.59). 3D method showed the best performance when compared to gold standard, exhibiting high sensitivity and specificity, therefore, it was validated as a reliable method to evaluate pit and fissure sealants

Sedative Effect Of Oral Diazepam And Chloral Hydrate In The Dental Treatment Of Children

Purpose: The purpose was to evaluate two sedation protocols during dental sessions in anxious children. Materials and Methods: It was a randomized and double-blind study, with each individual being his/her own control within each protocol. Furthermore, the two protocols were compared. Twenty children (36 to 84 months old) who exhibited "definitely negative" behavior according to the Frankl scale were assigned to receive oral chloral hydrate (40 mg/kg) (Group I) or Diazepam® (5 mg) (Group II). Behavior during local anesthesia, application of rubber dam, cavity preparation, restorative procedures was evaluated, considering the degree of sleep, body movement, crying and overall behavior. Vital signs were assessed at three different times. The Wilcoxon, Mann-Whitney, Exact Fisher's and Spearman correlation tests were used to analyze the data. Results: Group I presented higher scores for sleep during the CH session than placebo session during rubber dam application (P = 0.0431) and restoration (P = 0.0431). In Group II there was no statistically significant difference (p > 0.05). There were no statistically significant differences between sessions and groups in the evaluation of body movement, crying and vital signs. Overall behavior in the placebo session was better than in the CH session during local anesthesia, but there was no difference between the two drug regimens. There was influence of age during anesthesia and cavity preparation in Group I and during rubber dam application in Group II. It was concluded that oral diazepam and chloral hydrate had no influence on the behavior management for dental treatment with the studied sample.2526975Leelataweedwud, P., Vann Jr., W.F., Adverse events and outcomes of conscious sedation for pediatric patients: Study of an oral sedation regimen (2001) J Am Dent Assoc, 132, pp. 1531-1539American Academy of Pediatrics. Guidelines for monitoring and management of pediatric patients during and after sedation for diagnostic and therapeutic procedures: Addendum (2002) Pediatrics, 110, pp. 836-838. , Committee on DrugsDuncan, W.K., Pruhs, R.J., Ashrafi, M.H., Post, A.C., Chloral hydrate and other drugs used in sedating young children: A survey of American Academy of Pedodontics Diplomates (1983) Pediatr Dent, 5, pp. 252-256Klingberg, G., Berggren, U., Noren, J.G., Dental fear in an urban Swedish child population: Prevalence and concomitant factors (1994) Community Dent Health, 11, pp. 208-214Holst, A., Crossner, C.G., Direct ratings of acceptance of dental treatment In Swedish children (1987) Community Dent Oral Epidemiol, 15, pp. 258-263Klingberg, G., Vannas Lofqvist, L., Bjarnason, S., Noren, J.G., Dental behavior management problems in Swedish children (1994) Community Dent Oral Epidemiol, 22, pp. 201-205Badalaty, M.M., HouptMl, Koenigsberg, S.R., Maxwell, K.C., DesJardins, P.J., A comparison of chloral hydrate and diazepam sedation in young children (1990) Pediatr Dent, 12, pp. 33-37McCann, W., Wilson, S., Larsen, P., Stehle, B., The effects of nitrous oxide on behavior and physiological parameters during conscious sedation with a moderate dose of chloral hydrate and hydroxyzine (1996) Pediatr Dent, 18, pp. 35-41Reeves, S.T., Wiedenfeld, K.R., Wrobleski, J., Hardin, C.L., Pinosky, M.L., Arandomized double-blind trial of chloral hydrate/hydrazine versus midazolam/acetaminophen in the sedation of pediatric dental outpatients (1996) ASDC J Dent Child, 63, pp. 95-100Wilson, S., Easton, J., Lamb, K., Orchardson, R., Casamassimo, P., A retrospective study of chloral hydrate, meperidine, hydroxyzine and midazolam regimens used to sedate children for dental care (2000) Pediatr Dent, 22, pp. 107-112Jensen, B., Matsson, L., Benzodiazepines in child dental care: A survey of its use among general practitioners and paediatric dentists in Sweden (2001) Swed Dent J, 25, pp. 31-38Myers, G.R., Maestrello, C.L., Mourino, A.P., Best, A.M., Effect of submucosal midazolam on behavior and physiologic response when combined with oral chloral hydrate and nitrous oxide sedation (2004) Pediatr Dent, 26, pp. 37-43Cote, C.J., Sedation for the pediatric patient. A review (1994) Pediatr Clin North Am, 41, pp. 31-58zathan, J.E., Managing behavior of pre-cooperative children (1995) Dent Clin North Am, 39, pp. 789-816Malviya, S., Voepel-Lewis, T., Tait, A.R., Adverse events and risk factors associated with the sedation of children by nonanesthesiologists (1997) Anesth Analg, 85, pp. 1207-1213Milnes, A.R., Maupome, G., Cannon, J., Intravenous sedation in pediatric dentistry using midazolam, naibupine and droperidol (2000) Pediatr Dent, 22, pp. 113-119Wright GZ, Stanley PE, Garner DE. Minor sedation for apprehensive young children. In: Wright GZ, Stanley PE, Garner DE. Managing children's behavior in the dental office. Mosby: St. Louis1983. Chap 19Henderson BN 2 nd, Triplett RG, Gage TW. Anxiolytic therapy. Oral and intravenous sedation. Dent Clin North Am 1994;38:603-17Folayan, M.O., Faponle, A., Lamikanra, A., Seminars on controversial issues. A review of the pharmacological approach to the management of dental anxiety in children (2002) Int J Paediatr Dent, 12, pp. 347-354Frankl, L., Hellman, I., (1962) Symposium on child analysis. The Ego's participation in the therapeutic alliance. Int J Psychoanal, 43, pp. 333-337Houpt M. Project USAP the use of sedative agents in pediatric dentistry: 1991 update. Pediatr Dent 1993;15:36-40Radis, F.G., Wilson, S., Griffen, A.L., Coury, D.L., Temperament as a predictor of behavior during initial dental examination in children (1994) Pediatr Dent, 16, pp. 121-127Lindsay, S.J., Yates, J.A., The effectiveness of oral diazepam in anxious child dental patients (1985) Br Dent J, 159, pp. 149-153Yanase, H., Braham, R.L., Fukuta, O., Kurosu, K., A study of the sedative effect of home-administered oral diazepam forthe dental treatment of children (1996) Int J Pediatr Dent, 6, pp. 13-17Croswell, R.J., Dilley, D.C., Lucas, W.J., Vann Jr, W.F., A comparison of conventional versus electronic monitoring of sedated pediatric dental patients (1995) Pediatr Dent, 17, pp. 332-339Giovannitti Jr., J.A., Regimens for pediatric sedation (1993) Compendium, 14 (1002), pp. 1004-1006Hasty, M.F., Vann Jr, W.F., Dilley, D.C., Anderson, J.A., Conscious sedation of pediatric dental patients: An investigation of chloral hydrate, hydroxyzine pamoate, and meperidine vs. chloral hydrate and hydroxyzine pamoate (1991) Pediatr Dent, 13, pp. 10-19Needleman, H.L., Joshi, A., Griffith, D.G., Conscious sedation of pediatric dental patients using chloral hydrate, hydroxyzine and nitrous oxide-a retrospective study of 382 sedations (1995) Pediatr Dent, 17, pp. 424-431Reinemer, H.C., Wilson, C.F., Webb, M.D., A comparison of two oral ketamine-diazepam regimens for sedating anxious pediatric dental patients (1996) Pediatr Dent, 18, pp. 294-300Breimer, D.D., Clinical pharmacokinects of hypnotics (1977) Clin Pharmacokinet, 2, pp. 93-109Concise International Chemical Assessment Document no. 25. IPCS2000, , http://www.inchem.org/documents/cicads/cicads/cicad25. htm, International Programme on Chemical Safety, Available from:, Last accessed on 2005 Jan 10Jensen B, Schroder U, Mansson U. Rectal sedation with diazepam or midazolam during extractions of traumatized primary incisors: A prospective, randomized, double-blind trial in Swedish children aged 1.5-3.5 years. Acta Odontol Scand 1999;57:190-4Houpt, M.I., Weiss, N.J., Koenigsberg, S.R., Desjardins, P.J., Comparison of chloral hydrate with and without promethazine in the sedation of young children (1985) Pediatr Dent, 7, pp. 41-46Poorman, T.L., Farrington, F.H., Mourino, A.P., Comparison of a chloral hydrate/hydroxyzine combination with and without meperidine in the sedation of pediatric dental patients (1990) Pediatr Dent, 12, pp. 288-29

Neuropilin controls endothelial differentiation by mesenchymal stem cells from the periodontal ligament

Background: Periodontal ligament (PDL) has been reported to be a source of mesenchymal stem cells (MSCs). New vascular networks from undifferentiated cells are essential for repair/regeneration of specialized tissues, including PDL. The current study aims to determine potential of CD105(+)-enriched cell subsets of periodontal ligament cells (PDLSCs) to differentiate into endothelial cell (EC)-like cells and to give insights into the mechanism involved. Methods: CD105(+)-enriched PDLSCs were induced to EC differentiation by endothelial growth medium 2 (EGM-2) for 3, 7, 14, and 21 days, with mRNA/protein levels and functional activity assessed by: 1) real-time polymerase chain reaction; 2) Western blotting; 3) fluorescence-activated cell sorting; 4) immunohistochemistry; 5) immunofluorescence; 6) matrigel; and 7) small interfering RNA assays. Results: Data analyses demonstrated that EGM-2 treated PDLSCs presented increased expression of EC markers, including: 1) CD105; 2) kinase domain-containing receptor; and 3) Ulex europaeus agglutinin 1, and were able to form cord/tube-like structures. Gene and protein expression analysis showed that neuropilin 2 (NRP2), a key factor for vascular development, was significantly downregulated during EC differentiation. NRP2 was constitutively expressed in mouse PDL tissues by immunohistochemistry analysis, and NRP2 knockdown in CD105(+)-enriched PDLSCs resulted in increased cord/tube-like structures in a matrigel assay. Conclusion: These findings demonstrated the potential of CD105(+)-enriched PDLSCs to support angiogenesis, and NRP2 as a pivotal factor regulating this process.877E138E147COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPES02426/09-

Membrane proteome characterization of periodontal ligament cell sets from deciduous and permanent teeth

Physiological roles for the periodontal ligament (PDL) include tooth eruption and anchorage, force absorption, and provision of proprioceptive information. Despite the advances in understanding the biology of PDL cells, there is a lack of information regarding the molecular signature of deciduous (DecPDL) and permanent (PermPDL) PDL tissues. Thus, the present study was designed to characterize the membrane proteome of DecPDL and PermPDL cells. Primary PDL cells were obtained (n = 6) and a label-free quantitative proteome of cell membrane-enriched components was performed. Proteome findings were validated by quantitative polymerase chain reaction and Western blot assays in fresh human tissues (n = 8) and primary cell cultures (n = 6). In addition, confocal microscopy was used to verify the expression of target factors in the PDL cell cultures. Comparative gene ontology enrichment analysis evidenced that most stickling differences involved "endomembrane system" (PICALM, STX4, and LRP10), "hydrolase activity" (NCSTN and XRCC6), "protein binding" (PICALM, STX4, GPNMB, VASP, extended-synaptotagmin 2 [ESYT2], and leucine-rich repeat containing 15 [LRRC15]), and "isomerase activity" (FKBP8). Data are available via ProteomeXchange with identifier PXD010226. At the transcript level, high PICALM in DecPDL and ESYT2 and LRRC15 in PermPDL were confirmed in fresh PDL tissues. Furthermore, Western blot analysis confirmed increased levels of PICALM, LRRC15, and ESYT2 in cells and/or fresh tissues, and confocal microscopy confirmed the trends for PICALM and LRRC15 expression in PDL cells. We report the first comprehensive characterization of the membrane protein machinery of DecPDL and PermPDL cells, and together, we identified a distinct molecular signature for these cell populations, including unique proteins for DecPDL and PermPDL907775787CNPQ - Conselho Nacional de Desenvolvimento Científico e TecnológicoFAPESP – Fundação de Amparo à Pesquisa Do Estado De São Paulo304680/2014-12016/13786-0; 2016/02942-1; 2015/06372-

Secretome profiling of periodontal ligament from deciduous and permanent teeth reveals a distinct expression pattern of laminin chains

It has been suggested that there are histological and functional distinctions between the periodontal ligament (PDL) of deciduous (DecPDL) and permanent (PermPDL) teeth. Thus, we hypothesized that DecPDL and PermPDL display differences in the constitutive expression of genes/proteins involved with PDL homeostasis. Primary PDL cell cultures were obtained for DecPDL (n = 3) and PermPDL (n = 3) to allow us to perform label-free quantitative secretome analysis. Although a highly similar profile was found between DecPDL and PermPDL cells, comparative secretome analysis evidenced that one of the most stickling differences involved cell adhesion molecules, including laminin subunit gamma 1 (LAMC1) and beta 2 (LAMB2). Next, total RNA and protein extracts were obtained from fresh PDL tissues of deciduous (n = 6) and permanent (n = 6) teeth, and Western blotting and qPCR analysis were used to validate our in vitro findings. Western blot analysis confirmed that LAMC1 was increased in DecPDL fresh tissues (p< 0.05). Furthermore, qPCR data analysis revealed that mRNA levels for laminin subunit beta 1 (LAMB1), beta 3 (LAMB3), LAMC1, and gamma 2 (LAMC2) were higher in DecPDL fresh tissues, whereas transcripts for LAMB2 were increased in PermPDL (p< 0.05). In conclusion, the differential expression of laminin chains in DecPDL and PermPDL suggests an involvement of laminin-dependent pathways in the control of physiological differences between them11

Secretome Profiling of Periodontal Ligament from Deciduous and Permanent Teeth Reveals a Distinct Expression Pattern of Laminin Chains

<div><p>It has been suggested that there are histological and functional distinctions between the periodontal ligament (PDL) of deciduous (DecPDL) and permanent (PermPDL) teeth. Thus, we hypothesized that DecPDL and PermPDL display differences in the constitutive expression of genes/proteins involved with PDL homeostasis. Primary PDL cell cultures were obtained for DecPDL (n = 3) and PermPDL (n = 3) to allow us to perform label-free quantitative secretome analysis. Although a highly similar profile was found between DecPDL and PermPDL cells, comparative secretome analysis evidenced that one of the most stickling differences involved cell adhesion molecules, including laminin subunit gamma 1 (LAMC1) and beta 2 (LAMB2). Next, total RNA and protein extracts were obtained from fresh PDL tissues of deciduous (n = 6) and permanent (n = 6) teeth, and Western blotting and qPCR analysis were used to validate our <i>in vitro</i> findings. Western blot analysis confirmed that LAMC1 was increased in DecPDL fresh tissues (p<0.05). Furthermore, qPCR data analysis revealed that mRNA levels for laminin subunit beta 1 (<i>LAMB1)</i>, beta 3 (<i>LAMB3)</i>, <i>LAMC1</i>, and gamma 2 (<i>LAMC2)</i> were higher in DecPDL fresh tissues, whereas transcripts for <i>LAMB2</i> were increased in PermPDL (p<0.05). In conclusion, the differential expression of laminin chains in DecPDL and PermPDL suggests an involvement of laminin-dependent pathways in the control of physiological differences between them.</p></div

Childhood Hypophosphatasia Associated with a Novel Biallelic ALPL Variant at the TNSALP Dimer Interface

The goal of this study was to perform a clinical and molecular investigation in an eight-year-old female child diagnosed with hypophosphatasia (HPP). The proband and her family were evaluated by medical and dental histories, biochemical analyses, radiographic imaging, and genetic analysis of the tissue-nonspecific alkaline phosphatase (ALPL) gene. A bioinformatic analysis was performed to predict the structural and functional impact of the point mutations in the tissue-nonspecific alkaline phosphatase (TNSALP) molecule and to define their potential contribution to the phenotype. We identified a novel combination of heterozygous ALPL missense variants in the proband, p.Ala33Val and p.Asn47His, compatible with an autosomal recessive mode of inheritance and resulting in skeletal and dental phenotypes. Computational modeling showed that the affected Asn47 residue is located in the coil structure close to the N-terminal &alpha;-helix, whereas the affected Ala33 residue is localized in the N-terminal &alpha;-helix. Both affected residues are located close to the homodimer interface, suggesting they may impair TNSALP dimer formation and stability. Clinical and biochemical follow-up revealed improvements after six years of ERT. Reporting this novel combination of ALPL variants in childhood HPP provides new insights into genotype&ndash;phenotype associations for HPP and specific sites within the TNSALP molecule potentially related to a childhood-onset HPP and skeletal and dental manifestations. Beneficial effects of ERT are implicated in skeletal and dental tissues

Summary of secretome analysis of the PDL cells harvested from deciduous and permanent teeth.

<p><b>(A)</b> Venn diagram of total proteins and their top enriched molecular function (MF) GO terms. The MF GO terms were generated by DAVID software based on biological processes (GO_TERM_MF_2 database). <b>(B)</b> Volcano plot analysis of deciduous and permanent PDL cells secretome. Regulated proteins are painted in blue and orange, for deciduous and permanent teeth, respectively (<i>p</i><0.05).</p