Os primeiros passos para a implementação de um banco de dentes humanos na Faculdade de Odontologia UFF / The first steps to implement a human teeth bank at Faculty of Dentistry - UFF

O Banco de Dentes Humanos (BDH) é uma instituição sem fins lucrativos, vinculada a uma faculdade, universidade ou instituição com o propósito de suprir as necessidades acadêmicas, através do fornecimento de dentes humanos para estudo e treinamento laboratorial dos alunos dos cursos de graduação e pós-graduação. Assim, o presente trabalho teve como objetivo relatar os métodos utilizados para captação, tratamento, seleção, preservação, armazenamento e reutilização de dentes humanos extraídos, realizados por alunos da Faculdade de Odontologia (FO) da Universidade Federal Fluminense (UFF) através de dois projetos deferidos junto a PROAES-UFF. Para isso, foram realizadas campanhas de conscientização e programas de divulgação para a doação de dentes humanos extraídos permanentes e decíduos tanto para a comunidade científica odontológica como para a comunidade leiga. Após 24 meses, foram captados um total de 2.536 dentes, sendo 346 molares superiores, 279 molares inferiores, 262 pré-molares superiores, 418 pré-molares inferiores, 343 caninos, 296 incisivos superiores e 592 incisivos inferiores. As campanhas e ações dos projetos da PROAES-UFF foram eficientes para coletar dentes humanos, suprindo as atividades laboratoriais de diversas disciplinas na graduação e também servindo de estoque para a futura implementação do BDH da FO-UFF.

The Human Teeth Bank (HTB) is a non-profit institution, linked to a college, university or institution with the purpose of meeting academic needs, through the provision of human teeth for study and laboratory training of undergraduate and postgraduate students. Thus, this study aimed to report the methods of capture, treatment, selection, preservation, storage and reuse of extracted human teeth, carried out by students from the Faculty of Dentistry (FO) of Universidade Federal Fluminense (UFF) through two projects approved by PROAES-UFF. To this end, awareness campaigns and outreach programs were carried out for the donation of human teeth extracted, permanent and deciduous, both for the dental scientific community and for the lay community. After 24 months, a total of 2,536 teeth were captured, with 346 upper molars, 279 lower molars, 262 upper premolars, 418 lower premolars, 343 canines, 296 upper incisors and 592 lower incisors. The campaigns and actions of PROAES-UFF projects were efficient in collecting human teeth, supplying the laboratory activities of several disciplines during graduation and also serving as a stock for a future implementation of the HTB of FO-UFF.

Human DNA extraction from whole saliva that was fresh or stored for 3, 6 or 12 months using five different protocols

Abstract Saliva when compared to blood collection has the following advantages: it requires no specialized personnel for collection, allows for remote collection by the patient, is painless, well accepted by participants, has decreased risks of disease transmission, does not clot, can be frozen before DNA extraction and possibly has a longer storage time. Objective and Material and Methods This study aimed to compare the quantity and quality of human DNA extracted from saliva that was fresh or frozen for three, six and twelve months using five different DNA extraction protocols: protocol 1 – Oragene™ commercial kit, protocol 2 – QIAamp DNA mini kit, protocol 3 – DNA extraction using ammonium acetate, protocol 4 – Instagene™ Matrix and protocol 5 – Instagene™ Matrix diluted 1:1 using proteinase K and 1% SDS. Briefly, DNA was analyzed using spectrophotometry, electrophoresis and PCR. Results Results indicated that time spent in storage typically decreased the DNA quantity with the exception of protocol 1. The purity of DNA was generally not affected by storage times for the commercial based protocols, while the purity of the DNA samples extracted by the noncommercial protocols typically decreased when the saliva was stored longer. Only protocol 1 consistently extracted unfragmented DNA samples. In general, DNA samples extracted through protocols 1, 2, 3 and 4, regardless of storage time, were amplified by human specific primers whereas protocol 5 produced almost no samples that were able to be amplified by human specific primers. Depending on the protocol used, it was possible to extract DNA in high quantities and of good quality using whole saliva, and furthermore, for the purposes of DNA extraction, saliva can be reliably stored for relatively long time periods. Conclusions In summary, a complicated picture emerges when taking into account the extracted DNA’s quantity, purity and quality; depending on a given researchers needs, one protocol’s particular strengths and costs might be the deciding factor for its employment.

Assessment of the quantity and purity of DNA obtained from buccal cells under different storage methods / Avaliação da quantidade e pureza do DNA obtido por meio de raspagem de células da mucosa buccal sob diferentes condições de armazenamento

Aim: This study aimed to verify the quantity and purity of DNA obtained from buccal cells under different storage conditions. Methods: Thirty students, between 18 and 23 years of age participated in the study. Three samples of genetic material were collected from each student (samples A, B, and C) through a mouth rinse with 5 mL of 3% glucose. The first phase of DNA extraction from sample A was carried out on the same day of sample collection, whereas samples B and C were stored in a refrigerator and a freezer, respectively, for 1 month before the first extraction phase. DNA extraction was performed with 10 M ammonium acetate and 1 mM EDTA. Sample purity was assessed by spectrophotometry. Statistical analyses were performed through descriptive analysis and analysis of variance ANOVA using the SPSS software, version 21.0. Results: the samples presented no statistically significant differences between the DNA quantity (p = 0.37) or quality (p = 0.16). Conclusion: the quantity and purity of DNA extraction from the three samples were satisfactory, and no differences in storage conditions were found.(AU)

Objetivo: O objetivo desta pesquisa foi avaliar a quantidade e pureza do DNA obtido por células bucais utilizando diferentes meios de armazenagem. Métodos: trinta estudantes do curso de Odontologia entre 18 e 23 anos participaram desta pesquisa. O material genético foi coletado 3 vezes de cada indivíduo (amostras A B e C) por meio de bochechos com 5 ml de glicose 3%. Para a amostra A, foi realizada a primeira fase da extração do DNA no dia da coleta, já as amostras B e C, ficaram armazenadas em geladeira e freezer, respectivamente, por um mês antes da primeira extração. A extração do DNA foi realizada com acetato de amônio 10M e EDTA 1mM. Avaliouse a pureza das amostras por espectrofotometria. Resultados: as amostras não apresentaram diferenças estatisticamente significativas entre a quantidade (p = 0,37) ou pureza (p = 0,16) do DNA. Conclusão: a quantidade e a pureza do DNA das três amostras foram satisfatórias e não houve diferenças nas condições de armazenamento.(AU)

The use of a DNA stabilizer in human dental tissues stored under different temperature conditions and time intervals

Objective: The present study evaluated the use of a reagent to stabilize the DNA extracted from human dental tissues stored under different temperature conditions and time intervals. Material and Methods: A total of 161 teeth were divided into two distinct groups: intact teeth and isolated dental pulp tissue. The samples were stored with or without the product at different time intervals and temperature. After storage, DNA extraction and genomic DNA quantification were performed using real-time PCR; the fragments of the 32 samples that represented each possible condition were analyzed to find the four pre-selected markers in STR analysis. Results: The results of the quantification showed values ranging from 0.01 to 10,246.88 ng/μL of DNA. The statistical difference in the quantity of DNA was observed when the factors related to the time and temperature of storage were analyzed. In relation to the use of the specific reagent, its use was relevant in the group of intact teeth when they were at room temperature for 30 and 180 days. The analysis of the fragments in the 32 selected samples was possible irrespective of the amount of DNA, confirming that the STR analysis using an automated method yields good results. Conclusions: The use of a specific reagent showed a significant difference in stabilizing DNA in samples of intact human teeth stored at room temperature for 30 and 180 days, while the results showed no justification for using the product under the other conditions tested. .