Análise de células-tronco adultas (CTA) em cultura de células de tecido epitelial de pequenos roedores (rodentia-stricognathi- sciurognathi)

As células-tronco adultas (CTA) são células multipotentes e não especializadas encontradas na medula óssea, no sangue periférico, na córnea, na retina, no cérebro, no músculo esquelético, na polpa dental, no fígado, no pâncreas, no epitélio da pele, no sistema digestivo, no cordão umbilical e na placenta. Estas células podem se renovar e reproduzir indefinidamente e, sob certos estímulos, se transformar em células especializadas de diferentes tecidos ou órgãos. O presente trabalho tem como objetivo a obtenção de CTA a partir de tecido epitelial de roedores silvestres de espécies diferentes (Oecomys concolor - um exemplar fêmea, Proechimys roberti - dois exemplares machos, Hylaeamys megacephalus - dois exemplares machos). A metodologia para isolamento e cultivo in vitro de amostras do tecido epitelial foi estabelecida, a partir de protocolos já descritos, avaliando aspectos morfológicos, estabilidade genômica, contagem e análise da viabilidade celular, potencial clonogênico e indução de diferenciação em osteócitos, condrócitos e adipócitos. Todas essas análises foram feitas pós-criopreservação das culturas. As CTA foram caracterizadas como população homogênea de células que proliferam in vitro, como células aderentes à superfície do plástico, tendo morfologia semelhante a fibroblastos e formato fusiforme, com alta taxa de crescimento e proliferação celular por várias passagens sucessivas, onde a autorrenovação celular foi avaliada por ensaios clonogênicos. Na análise para examinar a estabilidade genômica na P3, todas as amostras apresentaram cariótipo com número diplóide normal e estável. A metodologia empregada nos ensaios para diferenciação das CTA em linhagens osteogênica, condrogênica e adipogênica, apresentou resultados satisfatórios, onde as células mostraram a marcação desejada através das colorações Alizarin Red S, Alcian Blue e Oil Red O, respectivamente. Todas as amostras testadas apresentam capacidade de proliferação e diversidade de diferenciação, sendo potencialmente fornecedores de CTA provenientes da pele e podendo ser utilizados como organismos modelos de estudos em CT.

Avaliação da ponta de diamante CVC (Chemical Vapor Deposition) associada ao aparelho de ultra-som

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Potencial angiogênico de células pulpares humanas em hipóxia

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação histomorfológica da resposta da polpa de dentes de cães submetida ao capeamento com sistema adesivo, cimento de hidróxido de cálcio e dois tipos de agregado de trióxido mineral

Pós-graduação em Odontologia - FOA

Resposta de polpas capeadas com proteínas bioativas ou hidróxido de cálcio submetidas ou não, à ação do laser

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Cinética de expressão e localização de indutor de metaloproteínases (EMMPRIN) durante o desenvolvimento de lesão periapical induzida em camundongos

Pós-graduação em Odontologia - FOAR

Fotobiomodulação da expressão e produção de mediadores inflamatórios por células pulpares irradiada com LED infravermelho

Pós-graduação em Ciências Odontológicas - FOAR

Citotoxidade de cimentos resinosos sobre célular obtidas da polpa de dentes humanos (HDPCs) e odontoblastóides (MDPC-23)

Pós-graduação em Reabilitação Oral - FOAR

Biologia pulpar: da agressão à reparação

The study of the dental pulp can be extended from factors related to its aggression to those related to new concepts of regeneration. The purpose of this compilation of studies is to present the evolution of a research subject from damage to repair. Innitially, studies will demonstrate the ability of dental procedures to generate heat and consequently affect the dental pulp. In sequence, studies will also present some effects of different pulp capping materials on dental pulp cells, related to the cytotoxicity of these materials and inflammatory potential. Finally, as the subject is emmerging and gaining importance in the literature, this compilation will present data from recent studies on the role of dental pulp progenitor cells in the regeneration and repair of dental pulp, as well as an alternative for a scaffold that could be used for clinical translation of research in the field. In summary, dentists must be aware of these different aspects and that the knowledge on factors and mechanisms involved in the aggression of the dental pulp can also serve as basis for understanding aspects for regeneration.

In Vitro Regulation of CCL3 and CXCL12 by Bacterial By-products Is Dependent on Site of Origin of Human Oral Fibroblasts

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Bleaching effectiveness, hydrogen peroxide diffusion, and cytotoxicity of a chemically activated bleaching gel

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Novel Chemically Modified Bacterial Cellulose Nanocomposite as Potential Biomaterial for Stem Cell Therapy Applications

Bacterial cellulose (BC) has become established as a remarkably versatile biomaterial and can be used in a wide variety of applied scientific applications, especially for medical devices. In this work, the bacterial cellulose fermentation process is modified by the addition of hyaluronic acid and gelatin (1% w/w) to the culture medium before the bacteria is inoculated. Hyaluronic acid and gelatin influence in bacterial cellulose was analyzed using Transmission Infrared Spectroscopy (FTIR) and Scanning Electron Microscopy (SEM). Adhesion and viability studies with human dental pulp stem cells using natural bacterial cellulose/hyaluronic acid as scaffolds for regenerative medicine are presented for the first time in this work. MTT viability assays show higher cell adhesion in bacterial cellulose/gelatin and bacterial cellulose/ hyaluronic acid scaffolds over time with differences due to fiber agglomeration in bacterial cellulose/gelatin. Confocal microscopy images showed that the cell were adhered and well distributed within the fibers in both types of scaffolds.

Apical periodontitis and periodontal disease increase serum IL-17 levels in normoglycemic and diabetic rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Tissue-engineering-based Strategies for Regenerative Endodontics

Stemming from in vitro and in vivo pre-clinical and human models, tissue-engineering-based strategies continue to demonstrate great potential for the regeneration of the pulp-dentin complex, particularly in necrotic, immature permanent teeth. Nanofibrous scaffolds, which closely resemble the native extracellular matrix, have been successfully synthesized by various techniques, including but not limited to electrospinning. A common goal in scaffold synthesis has been the notion of promoting cell guidance through the careful design and use of a collection of biochemical and physical cues capable of governing and stimulating specific events at the cellular and tissue levels. The latest advances in processing technologies allow for the fabrication of scaffolds where selected bioactive molecules can be delivered locally, thus increasing the possibilities for clinical success. Though electrospun scaffolds have not yet been tested in vivo in either human or animal pulpless models in immature permanent teeth, recent studies have highlighted their regenerative potential both from an in vitro and in vivo (i.e., subcutaneous model) standpoint. Possible applications for these bioactive scaffolds continue to evolve, with significant prospects related to the regeneration of both dentin and pulp tissue and, more recently, to root canal disinfection. Nonetheless, no single implantable scaffold can consistently guide the coordinated growth and development of the multiple tissue types involved in the functional regeneration of the pulp-dentin complex. The purpose of this review is to provide a comprehensive perspective on the latest discoveries related to the use of scaffolds and/or stem cells in regenerative endodontics. The authors focused this review on bioactive nanofibrous scaffolds, injectable scaffolds and stem cells, and pre-clinical findings using stem-cell-based strategies. These topics are discussed in detail in an attempt to provide future direction and to shed light on their potential translation to clinical settings.

Doxycycline-Encapsulated Nanotube-Modified Dentin Adhesives

This article presents details of fabrication, biological activity (i.e., anti-matrix metalloproteinase [anti-MMP] inhibition), cytocompatibility, and bonding characteristics to dentin of a unique doxycycline (DOX)-encapsulated halloysite nanotube (HNT)-modified adhesive. We tested the hypothesis that the release of DOX from the DOX-encapsulated nanotube-modified adhesive can effectively inhibit MMP activity. We incorporated nanotubes, encapsulated or not with DOX, into the adhesive resin of a commercially available bonding system (Scotchbond Multi-Purpose [SBMP]). The following groups were tested: unmodified SBMP (control), SBMP with nanotubes (HNT), and DOX-encapsulated nanotube-modified adhesive (HNT+DOX). Changes in degree of conversion (DC) and microtensile bond strength were evaluated. Cytotoxicity was examined on human dental pulp stem cells (hDPSCs). To prove the successful encapsulation of DOX within the adhesivesbut, more important, to support the hypothesis that the HNT+DOX adhesive would release DOX at subantimicrobial levelswe tested the antimicrobial activity of synthesized adhesives and the DOX-containing eluates against Streptococcus mutans through agar diffusion assays. Anti-MMP properties were assessed via -casein cleavage assays. Increasing curing times (10, 20, 40 sec) led to increased DC values. There were no statistically significant differences (p > .05) in DC within each increasing curing time between the modified adhesives compared to SBMP. No statistically significant differences in microtensile bond strength were noted. None of the adhesives eluates were cytotoxic to the human dental pulp stem cells. A significant growth inhibition of S. mutans by direct contact illustrates successful encapsulation of DOX into the experimental adhesive. More important, DOX-containing eluates promoted inhibition of MMP-1 activity when compared to the control. Collectively, our findings provide a solid background for further testing of encapsulated MMP inhibitors into the synthesis of therapeutic adhesives that may enhance the longevity of hybrid layers and the overall clinical performance of adhesively bonded resin composite restorations.