RESUMOS APRESENTADOS
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FC028 - Fórum Científico
Área: 10 - Implantodontia básica e biomateriais
Apresentação: 07/09 - Horário: 13h30 às 17h00 - Sala: Cerejeira
Respostas biologicas de superfícies dentais e implantes no ambiente oral: perfil proteômico e microbioma por modelos in vitro e in vivo
Rodrigo Fernando E. Souza Martins, Magda Aline Nagasawa, Raphael Cavalcante Costa, Magda Feres, Luciene Cristina de Figueiredo, Valentim Adelino Ricardo Barão, Jamil Awad Shibli, João Gabriel Silva Souza
UNIVERSIDADE GUARULHOS
Conflito de interesse: Não há conflito de interesse
A adsorção de proteínas em superfícies é considerada a primeira resposta biológica do corpo humano, sendo responsável por governar subsequentes processos biológicos, incluindo a adesão microbiana e processos inflamatórios. Além das superfícies dentais (esmalte e dentina), qualquer substrato inserido no ambiente bucal é imediatamente exposto à adsorção de proteínas e adesão microbiana, como as superfícies de implantes à base de titânio (Ti). Portanto, este estudo teve como objetivo comparar os perfis proteômicos e microbiológicos de Ti, dentina e esmalte após exposição à saliva e durante a inflamação induzida clinicamente. Utilizou-se cromatografia líquida acoplada à espectrometria de massas em tandem e a técnica de sequenciamento de RNA 16S. Nos achados in vitro, dentre as 349 proteínas adsorvidas da saliva, algumas eram exclusivas do titânio (12), dentina (42) ou esmalte (3). Análises matemáticas mostraram maior similaridade do perfil proteômico do Ti e do esmalte, diferindo da dentina. Diferenças significativas foram observadas nos dados microbiológicos entre os substratos. A análise de correlação identificou proteínas que promoveram ou inibiram a adesão de bactérias específicas. Nos achados clínicos, envolvendo 16 pacientes com inflamação induzida, as superfícies dentais e de implantes mostraram certa similaridade no perfil proteômico associado à inflamação, com algumas diferenças na composição microbiológica, principalmente na fase de inflamação ativa.
Conclui-se que, as superfícies dentais e de implantes diferem nas respostas biológicas no ambiente bucal. Tais dados podem ser de suma importância no desenvolvimento de novos biomateriais com resposta biológica aprimorada.
FC029 - Fórum Científico
Área: 10 - Implantodontia básica e biomateriais
Apresentação: 07/09 - Horário: 13h30 às 17h00 - Sala: Cerejeira
Arcabouços de DNA funcionalizados com amelotina atuam no reparo ósseo de defeito crítico de ratos
Naara Gabriela Monteiro, Odir Nunes de Oliveira Filho, Nadeen Meshry, Ana Cláudia Ervolino da Silva, Ryan Lee Chan, Karina Midori Mori Carneiro, Roberta Okamoto
UNIVERSIDADE ESTADUAL PAULISTA - ARAÇATUBA
Conflito de interesse: Não há conflito de interesse
O presente trabalho possui o objetivo de caracterizar o reparo ósseo em defeito crítico de calvária preenchido com scaffolds de hidrogéis de DNA funcionalizados ou não com amelotina. 40 ratos, adultos jovens, machos, foram divididos em 5 grupos experimentais: CLOT (cirurgia de defeito crítico e preenchimento do defeito com coágulo), AUTO PT (cirurgia de defeito crítico e preenchimento do defeito com osso autógeno particulado), BUFFER (cirurgia de defeito crítico e preenchimento do defeito com o veículo do hidrogel), DNA (cirurgia de defeito crítico e preenchimento do defeito com scaffolds de DNA), AMTN (cirurgia de defeito crítico e preenchimento do defeito com scaffolds de DNA funcionalizados com amelotina). Foi realizado o defeito crítico em calvária, unilateral, com 5 mm de diâmetro. A eutanásia dos animais foi realizada 10 dias após o procedimento cirúrgico. As amostras foram destinadas para as análises de microtomografia computadorizada (micro-CT), histológica e imunoistoquímica buscando caracterizar a expressão de proteínas como: OCN, COL-1 e TRAP. Todos os dados quantitativos foram submetidos ao teste de homocedasticidade e o nível de significância foi em 0<0,05. A imunoistoquímica mostrou através das proteínas avaliadas que os arcabouços de DNA atuaram na dinâmica óssea reparacional pois induziram a neocolagênese e mineralização da matriz extracelular e em contrapartida houve expressão de TRAP, proteína envolvida na reabsorção do tecido ósseo.
Contudo, os resultados preliminares apontam que os arcabouços com scaffolds de DNA e scaffolds de DNA funcionalizados com amelotina são biomateriais promissores para a regeneração óssea e podem atuar positivamente no reparo ósseo de defeitos críticos em calvárias de ratos.
(Apoio: FAPs - FAPESP N° 22/08746-0)
HA001 - Hatton
Área: 1 - Biologia craniofacial
Apresentação: 05/09 - Horário: 08h00 às 11h30 - Sala: Seringueira
Effect of microtopographic priming of mesenchymal stem cells expressing BMP-9 on their osteogenic potential
Hiskell Francine Fernandes e Oliveira, Robson Diego Calixto, Jessica Frith, Márcio Mateus Beloti, Adalberto Luiz Rosa
Bone Research Lab UNIVERSIDADE DE SÃO PAULO - RIBEIRÃO PRETO
Conflito de interesse: Não há conflito de interesse
Mesenchymal stem cells CRISPR-Cas9 edited to express bone morphogenetic protein 9 (MSCsBMP9) can repair bone tissue, at least in part, due to osteoblastic differentiation. Here, MSCsBMP9 were primed by culturing on a microtopographic surface to increase their osteogenic potential. MSCsBMP9 were cultured on either a tissue culture plastic surface of micropillars with 5 μm width/spacing and 5 μm height (pMSCsBMP9) or a standard culture plastic (MSCsBMP9) in non-osteogenic medium that was alpha-MEM supplemented with 10% fetal bovine serum (FBS) and antibiotics for 5 days. BMP9 gene and protein expressions were evaluated by RT-PCR and ELISA, respectively. Osteoblastic differentiation was evaluated by gene expression of the bone markers osterix, alkaline phosphatase (Alp), bone sialoprotein, osteocalcin, and osteopontin by RT-PCR, and in situ ALP activity by fast red staining. Data were compared by t-test (p≤0.05). Gene (p=0.006) and protein expressions of BMP9 (p=0.001) were higher in pMSCsBMP9. Gene expression of all evaluated genes (p=0.001 for all of them but osteocalcin, p=0.002), and ALP activity (p=0.003) were all higher in pMSCsBMP9.
These findings show that priming MSCsBMP9 by microtopographic surface increased BMP9 expression and consequently their osteoblastic differentiation even in the absence of osteogenic medium as a consequence of the increase in BMP9 expression. Such insights could make way for exploring cellular therapy based on priming MSCsBMP9.
(Apoio: FAPs - FAPESP N° 2023/15859-9; 2023/01309-7)
HA002 - Hatton
Área: 1 - Cirurgia bucomaxilofacial
Apresentação: 05/09 - Horário: 08h00 às 11h30 - Sala: Seringueira
3D-printed bone grafts and plates enhance bone repair via proteoglycan promotion in rabbit premaxillary defects
Eduardo Dallazen, Gabrielle Das Graças Domingues Dias, Gustavo Ribeiro Ferreira, Sybele Saska, Jamil Awad Shibli, Edilson Ervolino, Afsar Naqvi, Leonardo Perez Faverani
Departamento de Diagnóstico e Cirurgia UNIVERSIDADE ESTADUAL PAULISTA - ARAÇATUBA
Conflito de interesse: Não há conflito de interesse
Critical bone defects pose a challenge in oral and maxillofacial surgery. Advancing additive manufacturing is crucial for personalized grafts and fixation materials, customizing treatments to individual needs, and enhancing bone healing. This study investigated the effectiveness of additive manufacturing (AM) for fixation plates and bone grafts in a rabbit model. Male rabbits underwent premaxillary osteotomies, then were divided into four groups: conventional plates and screws (PLATES), 3D-printed plates (3D-PRINTED PLATES), custom AM-fabricated bone grafts (GRAFTS), and a combination of both (PLATES+GRAFTS). Pain behavior assessment, mass spectrometry, western blotting, and micro computed tomography (MicroCT) were performed. Pain scores decreased over time with no inter-group differences (P>0.05). Mass spectrometry identified 120 proteins and 63 of them were higher expressed in test groups(P<0.05). Bone-related proteins were higher in test groups, validated by western blotting (P<0.05). Proteoglycans such as decorin and biglycan showed significant expression variations, indicating their role in bone repair and extracellular matrix restructuring. Micro-CT analysis revealed higher bone volume (BV/TV) and improved trabecular bone (Tb) parameters (number and thickness) in groups with AM-produced materials (3D-PRINTED PLATES and PLATES+GRAFTS) compared to the control group (P<0.05).
Overall, this study demonstrates the potential of additive manufacturing techniques in enhancing bone healing outcomes and elucidates the functions of proteoglycans in bone repair. These findings provide valuable insights for developing innovative strategies in regenerative medicine for bone defects.
(Apoio: CAPES N° 88887.716820/2022-00 | FAPs - Fapesp N° 2023/16726-2)
HA003 - Hatton
Área: 1 - Cirurgia bucomaxilofacial
Apresentação: 05/09 - Horário: 08h00 às 11h30 - Sala: Seringueira
Influence of Menstrual Cycle, Surgical Factors, Anxiety, and Genetics on Discomfort Perception during Third Molar Extraction
Giselle Emilaine da Silva Reis, Laís O´´ Hara Zazula, Bruna da Silva, Aline Monise Sebastiani, Juliana Feltrin de Souza, Yasmine Mendes Pupo, Delson João da Costa, Rafaela Scariot
Estomatologia UNIVERSIDADE FEDERAL DO PARANÁ
Conflito de interesse: Não há conflito de interesse
The aim of this cross-sectional observational study was to evaluate the patient's perception of surgical discomfort in third molar extraction and its association with individual, women's health, surgical, genetic, and anxiety-related variables. This study, approved by the ethics committee #43894621.3.0000.0102, was conducted with 200 women aged between 18 and 45 years at the Federal University of Paraná over two years. The intensity of surgical discomfort was assessed using the QCirDental questionnaire. Data on individual, women's health, and surgical variables were also cataloged. Polymorphisms in the genes COMT, SLC6A4, TRPV1, HTR2A, ESR1, and ESR2 were investigated, obtained through salivary DNA collection. Serum levels of progesterone, estradiol, and follicle-stimulating hormone were obtained from blood collection. Anxiety was assessed by the IDATE questionnaire. The data were subjected to statistical analysis with a significance level of 5%. Women with high body mass index (p=0.042), follicular phase of the menstrual cycle (p=0.045), fewer children (p=0.047), longer duration of surgical procedure (p=0.012), traumatic experience in dental surgery (p=0.021), use of anxiety medication (p=0.016), elevated IDATE levels (p=0.001), TT genotype for the rs174675 polymorphism in COMT in the additive and dominant model (p=0.014) and (p<=0.01) respectively, GG genotype for the rs6113 polymorphism in HTR2A in the dominant model (p=0.038) were variables associated with a higher perception of discomfort during tooth extraction.
Our findings emphasizes the need for individualized treatment. In clinical practice, approaches aimed at alleviating patient anxiety should be reinforced.
(Apoio: CAPES)
HA004 - Hatton
Área: 2 - Terapia endodôntica
Apresentação: 05/09 - Horário: 08h00 às 11h30 - Sala: Seringueira
Immunomodulatory properties of bioactive materials for dental pulp regeneration
Alice Corrêa Silva-Sousa, Cristiane Miranda França, Avathamsa Athirasala, Anissa Bartolome, Anthony Tahayeri, Luiz Eduardo Braga Bertassoni, Francisco Wanderley Garcia de Paula-silva
Odontologia Restauradora UNIVERSIDADE DE SÃO PAULO - RIBEIRÃO PRETO
Conflito de interesse: Não há conflito de interesse
Bioactive materials and the immune system interactions can lead to regeneration, inflammation, or fibrosis. In Endodontics, a promising regenerative strategy is to fill the pulp space with a resorbable hydrogel and seal the tooth with a bioactive cement. However, the immunomodulatory properties of the combination of such materials are unclear. Because they are used in together to regenerate the dental pulp, we aim to evaluate immunomodulatory properties of dental cements and gelatin methacryloyl (GelMA) on dental pulp cells and macrophages. Thus, Biodentine, MTA, Ca(OH)2, and Ketac Molar extracts were placed in contact with macrophages for 24 hours, then secretome was collected and tested in dental pulp cells (DPC) for 7 days. Both macrophages and DPC were tested for viability and gene expression. Groups were compared using ANOVA followed by Tukey's test (⍺= 5%). Bioactive materials did not affect macrophage or DPC viability, but inhibited the expression of tumor necrosis factor. Macrophages' secretome induced RUNX2 and SPP1 up-regulation. Next, different types of GelMA were evaluated for mechanical properties. GelMA with a stiffness range matching that of dental pulp (2 to 25 kPa) was selected for 3D printing as an array featuring a stiffness gradient. Macrophages were seeded on the arrays for 3 days, then tested for polarization into M1 (pro-inflammatory) or M2 (pro-regenerative). Soft GelMA induced M1 or M2 phenotype, depending on the stimuli added. Stiff hydrogel allowed M1 polarization solely.
Our results show that macrophage secretome induced expression of mineralization genes in DPC and that GelMA stiffness can induce M2 polarization. These results pave the way for a more comprehensive strategy for dental pulp regeneration.
(Apoio: CAPES N° 33002029032P4 | CAPES N° 88887.716956/2022-00 | NIH N° 1K01DE030484-01)
HA005 - Hatton
Área: 3 - Controle de infecção / Microbiologia / Imunologia
Apresentação: 05/09 - Horário: 08h00 às 11h30 - Sala: Seringueira
The search for novel biomarkers for the diagnosis of peri-implant diseases using proteomic approaches
Lais de Paula Sumback Sivila Souza, Giuliano Portolese Svesutti Cesar, Debora Reis Dias, Paula Aline Zanetti Campanerut-sá, Nikolaos Donos, Elena Calciolari, Mauricio Guimarães Araújo, Flavia Matarazzo
Odontologia UNIVERSIDADE ESTADUAL DE MARINGÁ
Conflito de interesse: Não há conflito de interesse
The aim of this prospective observational study was twofold: i) to determine the proteomic composition of both peri-implant crevicular fluid (PICF) and saliva collected from subjects diagnosed as having peri-implant mucositis and peri-implantitis, and ii) to analyze differences in their proteome profiles in an attempt to establish putative biomarker signatures that may assist in the discrimination, and possibly the prediction, of peri-implant diseases. Patients with dental implants were clinically and radiographically evaluated and categorized as having either peri-implant mucositis or peri-implantitis, based on implant sites presenting the worst clinical condition. PICF and saliva samples were collected and analyzed by LC-MS/MS. Interpretation of proteomic data was performed using the Ingenuity Pathway Analysis (IPA). Thirty-one samples of PICF and 44 samples of saliva from patients with peri-implant mucositis, and 15 samples of PICF and 16 samples of saliva from patients with peri-implantitis were analyzed. A total of 481 human proteins were quantified in PICF and 454 were quantified in saliva. Of these, 18 proteins in PICF and 96 proteins in saliva had a statistically significant difference between the two conditions.
These preliminary results suggest that the pool of proteins identified may be unique to each clinical peri-implant condition. Proteomics research can bring new insights into the pathogenesis and progression of peri-implant disease. Nonetheless, more detailed analysis is still required before we can define putative biomarkers for the disease.
(Apoio: CAPES N° 88887713631202200)
HA006 - Hatton
Área: 3 - Controle de infecção / Microbiologia / Imunologia
Apresentação: 05/09 - Horário: 08h00 às 11h30 - Sala: Seringueira
Methylene blue-loaded liposomal nanocarriers for photodynamic therapy targeted to Candida auris biofilm
Patrícia Michelle Nagai de Lima, Akram Abbasi, Veronica Lamastro, Maíra Terra Garcia, Paulo Henrique Fonseca do Carmo, Anita Shukla, Thainá Lopes Bueno, Juliana Campos Junqueira
Biociências e Diagnóstico Bucal INSTITUTO DE CIÊNCIA E TECNOLOGIA / ICT-UNESP-SJC
Conflito de interesse: Não há conflito de interesse
Candida auris is an emerging species with a high profile of resistance to antifungals. Photodynamic therapy (PDT) combines a photosensitizer (PS) with light irradiation to kill microorganisms. However, the most used PS, Methylene Blue (MB), shows limited penetration on Candida biofilms. The goal was to develop liposomes as nanocarriers to enhance the penetration of MB within C. auris biofilm in the PDT. For this, positively charged (MB-P) and negatively charged (MB-N) liposomes were fabricated by the thin-film hydration method and extrusion. Dynamic light scattering (DLS) was used to obtain hydrodynamic diameter, polydispersity index (PDI), and zeta-potential. The encapsulation efficiency (EE) was determined by liposomes lysing, and drug loading (DL) capacity by weighing the lyophilized sample. Next, the penetration of MB-liposomes within C. auris biofilm was evaluated by confocal microscopy. The efficacy of MB-P or MB-N irradiated with light-emitting diode (LED) on biofilms was assessed by the quantification of viable cells, total biomass, and reactive oxygen species (ROS). The biocompatibility of MB-P and MB-N on fibroblasts were also evaluated using a Cell Counting Kit-8 assay. The results showed that both MB-P and MB-N liposomes were successfully fabricated with a hydrodynamic diameter of 188 and 227 nm, PDI of 0.12 and 0.18 and zeta-potential of 28 and -22 mV, respectively. The EE was found to be 10 and 14%, and DL capacity of ~5 and 5.9%. MB-P and MB-N showed enhanced penetration within biofilms, resulting in significative increase of ROS production and reduction of biofilm cells compared to free MB. MB-P and MB-N maintained fibroblasts viability at 80%.
PDT using MB-P and MB-N can be an adjuvant treatment for C. auris infections.
(Apoio: CAPES N° 88887.696595/2022-00 | CAPES N° 88887.926029/2023-00 | CNPq N° 407032/2023-1)
HA007 - Hatton
Área: 3 - Fisiologia / Bioquimica / Farmacologia
Apresentação: 05/09 - Horário: 08h00 às 11h30 - Sala: Seringueira
Role of the succinate/Sucnr1 signaling pathway in TrkB positive neurons in the development of paclitaxel-induced neuropathic pain
Francisco Isaac Fernandes Gomes, Rafaela da Silva Castro, Gabriel Victor Lucena da Silva, Beatriz Lima Adjafre, Conceição Elidianne Aníbal Silva, Antonio Edson Rocha Oliveira, Isadora Marques Paiva, Thiago Mattar Cunha
Departamento de Farmacologia FACULDADE DE MEDICINA DE RIBEIRÃO PRETO - UNIVERSIDADE DE SÃO PAULO
Conflito de interesse: Não há conflito de interesse
We studied the role of the succinate/Sucnr1 pathway in primary sensory neurons (PSN) in the development of paclitaxel (PTX)-induced neuropathic pain, which was induced in mice by PTX (8mg/kg; i.p.), and succinate plasma levels were quantified (LC/MS). RNA-seq database reanalyzes determined the PSN gene signature post-PTX. Nociception was studied in wildtype (WT) and full knockout mice for Sucnr1 (Sucnr1-/-) after PTX, as well as, in WT mice and Sucnr1-/-, or celecoxib-treated mice (20mg/kg; i.p.), or mice lacking TNFR1/R2, IL6, ILR1, Caspase1, TRPA1, TRPV1, or conditional knockout (cKO) of Sucnr1 in dendritic cells after succinate (1nmol/paw). Neuronal effects were assessed by measuring firing rates post-succinate (0.3-10mM) in multielectrode array systems. Identification of Sucnr1-expressing PSN was achieved by RNA-seq database analyses and validated by RNA scope. Mice devoid of nociceptors and cKO of Sucnr1 in Nav1.8 PSN underwent succinate or PTX treatment for behavioral and molecular analyses. Data were analyzed by ANOVA followed by Bonferroni´s test or Student´s t-test (α=5%). PTX increased plasma levels of succinate and caused metabolic perturbation in PSN. Sucnr1-/- mice showed attenuated nociception after PTX. Succinate enhanced nociception in WT mice, but not in Sucnr1-/- mice. Yet, pharmacological, or genetic interventions did not change its effects. Succinate rendered PSN hyperexcitable at 10mM. TrkB neurons represented 55% of Sucnr1-expressing PSN. Pharmacological and genetic ablation did not change sensitization elicited by succinate or PTX-treatment, nor the expression of Sucnr1.
Thus, succinate/Sucnr1 pathway contributes to the development of PTX-induced neuropathic pain by sensitizing TrkB neurons.
(Apoio: FAPESP N° 2019/14285-3)
HA008 - Hatton
Área: 3 - Fisiologia / Bioquimica / Farmacologia
Apresentação: 05/09 - Horário: 08h00 às 11h30 - Sala: Seringueira
Non-invasive platform for Autism Spectrum Disorder detection based on Salivary Lipidomics coupled with Artificial Intelligence
Sttephany Silva Bernardino, Marjorie Adriane da Costa Nunes, Hebreia Oliveira Almeida-souza, Marco Fidel Guevara Vega, Arlene Bispo Dos Santos Nossol, Mario Machado Martins, Cyrene Piazera Silva Costa, Robinson Sabino-silva
UNIVERSIDADE FEDERAL DE UBERLÂNDIA
Conflito de interesse: Autodeclarado "Potencial depósito de patente INPI"
The timely and accurate diagnosis of Autism Spectrum Disorder (ASD) in clinical settings remains challenging due to the lack of diagnostic markers. The application of untargeted lipidomics has emerged as a powerful analysis to detect novel biomarkers. In this context, the non-invasive detection of salivary lipids can offer large-scale detection of ASD. In this study, we used Gas chromatography-mass spectrometry-based untargeted lipidomics coupled with artificial intelligence algorithms to identify changes in salivary lipid profile to be used as an alternative for ASD detection. Saliva samples from 33 ASD and matched-control children were analyzed. A total of 180 salivary lipids were identified, 31 had a frequency filter of 75% and some salivary peptides were pioneeringly identified. Three salivary lipids 1-Naphthalenamine, N-phenyl-; Hexadecanoic acid, 2-hydroxy-1-(hydroxymethyl)ethyl ester; and 13-Docosenamide, (Z)- were higher (p<0.05) expressed in ASD than matched-controls. We also tested 6 state-of-the-art Python-based artificial intelligence algorithms, the Neural Networks had the best performance achieving 84% of accuracy, 80% of specificity, and 88% of sensitivity with the highest SHAP score of 13-Docosenamide.
In summary, these data highlight the potential of salivary lipidomics profile supported by artificial intelligence algorithms as a non-invasive tool for screening of ASD children, enabling advances in precision medicine and personalized dentistry.
(Apoio: CNPq N° INCT Oral health and Dentistry #406840/2022-9 | CNPq N° INCT Theranostics and Nanobiotechnology 403193/2022-2 | CNPq N° Autism spectrum disorder #422205/2021-4)