HA001 - Hatton
Área: 2 - Biologia pulpar

Are all dental pulp stem cells primed to differentiate into vascular endothelial cells?
Bergamo, MTOP, Zhang Z, Oliveira TM, Machado MAAM, Nör JE
Odontopediatria, Ortodontia e Saúde Cole - UNIVERSIDADE DE SÃO PAULO - BAURU
Conflito de interesse: Não há conflito de interesse

The purpose of this study was to evaluate if there is a sub-population of dental pulp stem cells that is uniquely primed to differentiate into vascular endothelial cells. Here, we sorted SHED or DPSC according to expression levels of VEGFR1 and exposed them to one of the following conditions: 1) Control medium; 2) Endothelial differentiation medium - Endothelial Growth Medium (EGM) 2-MV supplemented with 50 ng/mL rhVEGF in presence of 0 or 25 μg/ml Bevacizumab. We evaluated cell proliferation and endothelial differentiation potential in vitro through the: Sulforhodamine-B (SRB) assay, Capillary Sprouting Assay, RT-PCR, Western Blot, and Immunofluorescence. We also transplanted VEGFR1-sorted cells into immunodeficient mice and evaluated the impact of VEGFR1 expression levels on microvessel density by histology and immunohistochemistry. We confirmed that dental pulp stem cells differentiate into endothelial cells by increased expression of endothelial cell markers and by capillary sprouting formation upon exposure to endothelial differentiation medium. VEGFR1^low dental pulp stem cells proliferated quicker than VEGFR1^high cells, indicating their preferential self-renewal. On the other hand, VEGFR1^high cells are more prone to vasculogenic differentiation, as they generated more capillary sprouts in vitro and more microvessels in vivo than VEGFR1^low cells.
These data demonstrate that stem cells of the dental pulp have an unique sub-population of cells defined by high VEGFR1 expression that are primed to differentiate into vascular endothelial cells.
(Apoio: FAPESP  N° 2018/13675-0  |  NIH/NIDCR   N° RO1-DE021410)

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HA002 - Hatton
Área: 3 - Fisiologia / Bioquimica / Farmacologia

Central antinociceptive action of Botulinum toxin type A and its effects on the glia-neuron cross-talk in a TMJ-rheumatoid arthritis model
Muñoz-Lora VRM, Abdalla HB, Matak I, Dugonjic A, Lackovic Z, Clemente-Napimoga JT, Cury AAB
Pós Graduação Em Odontologia - UNIVERSIDADE GUARULHOS
Conflito de interesse: Não há conflito de interesse

We assessed Botulinum toxin type A (BoNT/A) central enzymatic and antinociceptive activity and its possible influence on the glia-neuron communication using a model of antigen-evoked rheumatoid arthritis (RA) in the temporomandibular joint (TMJ) of rats. Wistar rats were induced to RA in the left TMJ by systemic and intra-articular (i.a) challenges using methylated bovine serum albumin. Then, animals were treated with i.a. unilateral (left TMJ) injection of BoNT/A. Spontaneous and mechanically-evoked nocifensive behaviors were assessed by behavioral nocifensive responses, rat grimace scale, and von Frey filaments. Animals were sacrificed and the trigeminal nucleus caudalis (TNC) was collected to perform: 1) immunohistochemical staining of cleaved SNAP25, c-Fos protein and glial fibrillary acidic protein (GFAP); 2) western blot of microglial P2X7 receptor and CX3 chemokine receptor 1 (CX3CR1); and 3) Elisa of microglial modulators Cathepsin S, Fractalkine and pro-inflammatory cytokines TNF-a and IL-1b. BoNT/A application was associated with the appearance of truncated SNAP-25 in the sensory TNC. BoNT/A reduced spontaneous and evoked nociceptive behaviors, bilateral c-Fos expression, and markers of neuronal and glial activation.
The antinociceptive activity of BoNT/A is associated with its ability to transport to upper sensory regions and reduce neuronal and glial cells activation. BoNT/A might be useful as a central acting and neuromodulatory drug for chronic pain conditions. However, BoNT/A effects on glial cells should be carefully explored.
(Apoio: FAPs - FAPESP  N° 2017/07741-7  |  FAPs - BEPE-FAPESP  N° 2018/13575-5  |  CAPES  N° 001)

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HA003 - Hatton
Área: 4 - Odontopediatria

Impact of biomineralization on resin/demineralized dentin bond longevity in a minimally invasive approach: an in vitro 18-month follow-up
Moreira KMS, Bertassoni LEB, Davies RPW, Joia F, Höfling JF, Nascimento FD, Puppin-Rontani RM
Odontologia Infantil - FACULDADE DE ODONTOLOGIA DE PIRACICABA
Conflito de interesse: Não há conflito de interesse

This study determined the impact of 0.2% NaF-NaF, casein phosphopeptide-amorphous calcium phosphate-CPP-ACP, or P₁₁-4 peptide on resin/dentin demineralized (DD) interface longevity. DD provided by biological method, used 255 caries-free third molars, randomly distributed into 5 groups: Sound dentin-SD; DD; DD+NaF-NaF; DD+CPP-ACP-CPP-ACP; DD+P₁₁-4-P₁₁-4. A block of Filtek^TM Z350 composite resin (4mm/height) was bonded with Adper^TM Single Bond 2 over dentin surfaces. The resin/dentin blocks stored in Simulated Body Fluid/37^oC under modified simulated pulpal pressure were tested by µTBS, nanoinfiltration, in situ zymography and µ-CT, at 24-h, 6 and 18-mo. Data from µTBS and µ-CT were submitted to ANOVA and Tukey tests, and from failure patterns and in situ zymography to Kruskal-Wallis test (α=5%). Descriptive analysis was performed for nanoinfiltration and hybrid layer formation/degradation. The highest µ-CT values were showed by CPP-ACP (p<0.01) and P11-4 (p<0.01) groups, over time; NaF showed similar values than DD (p>0.05), whereas CPP-ACP resembled the SD (p>0.05), over time; P11-4 reached values close to SD, at 24-h and 18-mo (p>0.05); DD and NaF showed highest adhesive failure all periods (p<0.05). SD, CPP-ACP and P11-4 presented the lowest nanoinfiltration and proteolytic activity, at the 24h and increased after 6 and 18-mo. The mineral density increased in the DD for CPP-ACP and P11-4 groups (p<0.05).
Therefore, CPP-ACP and P₁₁-4 showed to be a promising method for treating DD, increasing the longevity of adhesive restorations.

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HA004 - Hatton
Área: 4 - Odontopediatria

Proteomic profile of the acquired enamel pellicle of children with early childhood caries and caries-free children
Silva NC, Oliveira BP, Ventura TMO, Toniolo J, Buzalaf MAR, Rodrigues JA
UNIVERSIDADE FEDERAL DO RIO GRANDE DO SUL
Conflito de interesse: Não há conflito de interesse

Early Childhood Caries (ECC) is defined as the presence of one or more primary teeth with carious lesion on some surface, in children up to 6 years old. The acquired enamel pellicle (AEP) play an important role in the pathogenesis of ECC, working as a protective interface between the tooth surface and the oral cavity. There is no data on the proteomic profile of AEP from children with ECC. The objective of this study was to compare the proteomic profile of the in vivo AEP from children, aged 3 to 5 years old, with ECC (n=10) and caries-free (CF; n=10). After AEP samples have been collected, they were processed by proteomic analysis (nLC-ESI-MS/MS). For the label-free quantitative analysis the PLGS Software was used. In total, 241 proteins were identified. Among the exclusive proteins, basic salivary proline-rich protein (PRP) 1 and 2, cystatin-B and cystatin-SA were found only in CF and should be highlighted. When comparing ECC and CF, decreased proteins in the CF included 6 hemoglobin isoforms, serum albumin, neutrophil defensin 3 and proteins S100-A8 and A9, and increased proteins included submaxillary gland androgen-regulated protein 3B, histatin-1, statherin, 3 isoforms of PRP and alpha-amylase 1 and 2B.
These findings show that there are differences in the protein profile of AEP when compared ECC with CF children. The exclusive and the increased proteins found in CF group might have protective functions that play a role in the prevention to caries, besides provide important insights for the development of new therapeutic strategies and development of dental products for ECC.
(Apoio: CNPq  N° 314532/2018-8)

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HA005 - Hatton
Área: 4 - Ortodontia

Upper airway changes in miniscrew-anchored maxillary protraction with hybrid and hyrax expanders: a randomized clinical trial
Miranda F, Pugliese FS, Massaro C, Bastos JCC, Santos AM, Janson G, Palomo JM, Garib DG
Odontopediatria, Ortodontia e Sc - UNIVERSIDADE DE SÃO PAULO - BAURU
Conflito de interesse: Não há conflito de interesse

The aim of this study was to compare the upper airway space changes after miniscrew-anchored maxillary protraction with hybrid and conventional hyrax expanders. The sample comprised 40 Class III malocclusion growing patients that were randomized into two groups of miniscrew-anchored maxillary protraction. The group HH was treated a hybrid hyrax appliance in the maxilla and two miniscrews distally to the canines in the mandible. Class III elastics were used from the maxillary first molar to the mandibular miniscrews until anterior crossbite correction. The group CH was treated with a similar protocol except for the conventional hyrax expander in the maxilla. CBCT was obtained before (T1) and after 12 months of therapy (T2). The shape and size of upper airway were assessed. Intergroup comparisons were performed using t tests (P<0.05). The group HH was composed by 13 patients (6 female, 7 male) with a mean age of 10.42 years. The group CH was composed by 15 patients (5 female, 7 male) with a mean age of 11.38 years. Good reproducibility was found for all measurements. Anteroposterior and transverse increases of the upper airway were found for both groups. The oropharynx and the most constricted area increased similarly in both groups.
Maxillary protraction using miniscrews as anchorage produced an increase in the upper airways. No differences in upper airway changes were observed using protraction anchored on hybrid or conventional hyrax expanders.
(Apoio: FAPs - FAPESP  N° 2017/04141-9   |  FAPs - FAPESP  N° 2017/24115-2  |  FAPs - FAPESP  N° 2019/03175-2)

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