Biobancos en los servicios de anatomía patológica en Costa Rica: un análisis bioético y biojurídico

Objetivo: Analizar el marco bioético y biojurídico en relación con los biobancos en los servicios de anatomía patológica. Métodos: Revisión bibliográfica descriptiva de información relevante sobre la bioética y bioderecho en relación con los biobancos en los servicios de anatomía patológica, en donde se contemplaron las normativas, decretos y leyes nacionales e internacionales relacionadas con los biobancos y las bases de datos; se consultaron diferentes libros de texto utilizando el catálogo público del Sistema de Bibliotecas, Documentación e Información de la Universidad de Costa Rica y plataformas como PubMed y Google Académico. Resultados: Con el avance de la medicina en el desarrollo de nuevas técnicas terapéuticas y diagnósticas, surge una nueva herramienta de gran interés científico a nivel internacional: el establecimiento de biobancos de tejidos para conservar muestras biológicas utilizadas en estudios moleculares e investigación. Esto trajo consigo una cantidad de cuestionamientos de índole bioético y biojurídico relevantes relacionados con los derechos humanos y el respeto a la dignidad humana. Para establecer un biobanco en un departamento de anatomía patológica es indispensable conocer, manejar e implementar lo estipulado por las normativas nacionales e internacionales referente al consentimiento informado y la protección de los datos vinculados a los bioespecímenes. Conclusiones: Para utilizar en investigación los bioespecímenes que se encuentran en los departamentos de anatomía patológica debe seguirse lo que estipulan las normas y directrices nacionales e internacionales que regulan el manejo, uso y almacenamiento de las muestras, el consentimiento informado, la protección y la confidencialidad de los datos relacionados a las personas involucradas en los estudios biomédicos. En Costa Rica el marc ético-jurídico relacionado a la investigación biomédica regula el uso de los bioespecímenes en ensayos clínicos, pero no existe una legislación nacional específica en materia de biobancos, se recomienda que se establezca una ley con su respectivo reglamento en donde se visualicen todos los requerimientos de un biobanco a nivel público y privado.

Aim: To analyze the bioethical and bio juridical framework regarding biobanks in the Pathology Departments. Methods: Descriptive bibliographical revision of relevant information regarding bioethics and bio law with focus on the pathological anatomy services. Local laws, international laws and normative linked to biobanks and data bases were taken into consideration. Text books from the Universidad de Costa Rica`s Library, Documentation and Information System were consulted and also digital platforms as Academic Google and PubMed. Results: A new tool of scientific interest has emerged internationally with the medical advancement in the development of new therapeutic and diagnostic techniques: the establishment of tissue biobanks to preserve biological samples for molecular studies and research. This brought with it a number of questions relevant to bioethics and biolaw related to human rights and the respect for human dignity. In order to establish a biobank in a pathological anatomy department, it is essential to know, how to manage and implement the national and international regulations related to informed consent and the protection of data linked to biological samples. Conclusions: National and international guidelines for using biological samples stored in pathology departments for research purposes must be followed. They must regulate the management, use and storage of samples. Also, the informed consent and the protection and confidentiality of the data regarding the incumbent people in biomedical research must be take into consideration. In Costa Rica, the ethical and juridical framework related to the biomedical research establishes the parameters of biospecimens in clinical rehearsals but there is not a national legislation regarding biobanks. Because of that, is that is recommended a regulation that must include all the requirements for a biobank either in the public and private spectrum; such as a national law and formal regulation.

Biobancos: conservación del patrimonio científico de la humanidad

Un biobanco permite mantener en condiciones óptimas y de manera organizada las colecciones de muestras biológicas con los más altos estándares de calidad, éticos y legales, con fines de investigación biomédica que genere nuevo conocimiento, así como aplicaciones de docencia. Los avances en biomedicina y el desarrollo de tecnologías, han facilitado el acceso a la información y esto debe aplicarse en pro de un mejor aprovechamiento de las muestras biológicas, en la comprensión de las bases inmunopatogénicas de las enfermedades, que finalmente lleve a proponer estrategias de detección, prevención, promoción y tratamiento. El Laboratorio de Dermatopatología de la Sección de Dermatología de la Facultad de Medicina de la Universidad de Antioquia, ha procesado más de 57.000 biopsias desde su fundación, conservando los archivos clínicos, registros fotográficos, placas y bloques histopatológicos, otorgándole un gran potencial como biobanco. Esta revisión narrativa de la literatura tiene como objetivo ilustrar una visión general de la importancia del desarrollo de los biobancos, así como los aspectos a tener en cuenta en su estructuración.

Summary A biobank allows the collections of biological samples to be maintained in optimal conditions and in an organized manner with the highest quality, ethical and legal standards, for biomedical research purposes that generate new knowledge, as well as teaching applications. Advances in biomedicine and the development of technologies have facilitated access to information and this should comply in favor of a better use of biological samples in the understanding of the immunopathogenic bases of diseases, which finally carry out a proposing strategies of detection, prevention, promotion and treatment. The Dermatopathology Laboratory of the Dermatology Section of the Faculty of Medicine of the University of Antioquia has processed more than 57,000 biopsies since its foundation, preserving the clinical files, photographic records, plates and histopathological blocks, giving it great potential as a biobank. This narrative review of the literature aims to illustrate an overview of the importance of the development of biobanks, as well as the aspects to take into account in their structuring.

Situación actual de los bancos de tejidos en Colombia: tejido cardiovascular / An update on cardiovascular tissue banks in Colombia

Resumen Introducción: ¿Cuál es la situación actual técnica y productiva de los cuatro bancos que están certificados en buenas prácticas para tejido cardiovascular en Colombia si se comparan con estándares internacionales? Objetivo: Caracterizar la capacidad técnica y la actividad de los bancos certificados del país para tejido cardiovascular durante el periodo de tiempo 2014 a 2016. Metodología: Mediante una encuesta realizada por vía teleconferencia a cada banco, se evaluaron ítems de donación, extracción, procesamiento, almacenamiento, distribución y capacidad de producción de los tejidos cardiovasculares. La información referida por las instituciones fue complementada con actas de certificación de buenas prácticas del INVIMA y la suministrada por el INS de las estadísticas reportadas por los mismos bancos durante el periodo 2014 a 2016. Resultados: El tejido cardiovascular procesado por los bancos colombianos procede principalmente de donantes con muerte encefálica y es extraído directamente en el quirófano; la edad de los donantes va desde recién nacidos hasta los 60 años. Cuando se requiere, el antibiótico de elección es la vancomicina. El almacenamiento más usado es la criopreservación y su distribución se hace principalmente a grupos quirúrgicos preestablecidos e intrainstitucionales. Conclusiones: El desarrollo del banqueo de este tipo de tejido está muy rezagado en Colombia respecto a otros países, debido a las dificultades de donación, extracción y utilización por grupos con alta experticia; adicionalmente, la normativa utilizada para evaluar estas instituciones está muy desactualizada.

Abstract Introduction: A study is performed to determine if the current technical and productive situation of the four cardiovascular tissue banks that have Good Practice Certificates in Colombia compare with international standards. Objective: To determine the technical capacity and activity of the banks certified by the country for cardiovascular tissue during the period from 2014 to 2016. Methodology: The items analysed using a questionnaire completed by teleconference to each bank were, donation, extraction, processing, storage, distribution, and production capacity of the cardiovascular tissues. The information provided by the institutions was complemented with the Minutes of Good Practice Certification of the Colombia National Food and Drug Surveillance Institute (INVIMA) as well as the statistics reported by the banks themselves and provided by the National Health Institute (INS) during the period 2014 to 2016. Results: The cardiovascular tissue processed by Colombian tissue banks mainly comes from donors with brain death, and is extracted directly in the operating theatre. The age of the donors go from newborn up to 60 years-old. When used, the antibiotic of choice is vancomycin. The most used storage method is cryopreservation, and is mainly distributed to established and institutional surgical groups. Conclusions: The development of this type of tissue bank is lagging behind in Colombia compared to other countries. This is due to the difficulties in donation, extraction, and use by groups with high expertise. Furthermore, the guidelines used to evaluate these institutions are very out of date.

Characterization of Decellularized Human Pericardium for Tissue Engineering and Regenerative Medicine Applications / Caracterização do Pericárdio Humano Descelularizado para Engenharia de Tecidos e Aplicações de Medicina Regenerativa

Abstract Background: Pericardium tissue allograft can be used for surgical repair in several procedures. One of the tissue engineering strategies is the process of decellularization. This process decreases immunogenic response, but it may modify the natural extracellular matrix composition and behavior. Objective: The aim of this study was to evaluate the effectiveness of cell removal, maintenance of extracellular matrix properties and mechanical integrity of decellularized human pericardium using a low concentration solution of sodium dodecyl sulfate. Methods: Decellularization was performed with sodium dodecyl sulfate and ethylenediaminetetraacetic acid. Histological analysis, DNA quantification, evaluation of glycosaminoglycans and collagen were performed. Biomechanical assay was performed using tensile test to compare the decellularization effects on tissue properties of tensile strength, elongation and elastic modulus. P < 0.05 was considered significant. Results: There was reduction in visible nuclei present in pericardium tissue after decellularization, but it retained collagen and elastin bundles similar to fresh pericardium. The DNA contents of the decellularized pericardium were significantly reduced to less than 511.23 ± 120.4 ng per mg of dry weight (p < 0.001). The biomechanical assay showed no significant difference for fresh or decellularized tissue. Conclusion: The decellularization process reduces cell content as well as extracellular matrix components without changing its biomechanical properties.

Resumo Fundameto: O enxerto de pericárdio pode ser usado em muitos procedimentos de correção cirúrgica. Uma das estratégias da engenharia tecidual é o processo de descelularização. No entanto, embora esse processo diminua a resposta imunogênica, a descelularização pode modificar tanto o comportamento como a composição da matriz extracelular natural. Objetivos: Avaliar a eficácia da descelularização usando baixa concentração de dodecil sulfato de sódio na remoção celular, na manutenção das propriedades da matriz extracelular e na integridade mecânica do pericárdio humano descelularizado. Métodos: A descelularização foi realizada com dodecil sulfato de sódio e ácido etilenodiamino tetra-acético. Foi realizada análise histológica, quantificação de DNA, e avaliação de glicosaminoglicanos e colágeno. O estudo biomecânico foi conduzido pelo teste de tração para comparar os efeitos da descelularização sobre as propriedades teciduais de resistência à tração, alongamento e módulo de elasticidade. Foi considerado um valor de p < 0,05 como estatisticamente significativo. Resultados: Observou-se uma redução na quantidade de núcleos presentes no pericárdio após a descelularização, apesar de manter quantidades similares de feixes de elastina e de colágeno. As concentrações de DNA do pericárdio descelularizado foram significativamente reduzidas para menos que 511,23 ± 120,4 ng por mg de peso seco (p < 0,001). O teste biomecânico não apontou diferenças entre os tecidos fresco e descelularizado. Conclusão: A descelularização reduziu a concentração de células bem como os componentes da matriz extracelular sem afetar suas propriedades biomecânicas.

Distribuição do tecido ocular no Estado de São Paulo: análise por razões de descarte de córneas / Ocular tissue distribution in the State of São Paulo: analysis on corneal discarding reasons / Distribución del tejido ocular en el estado de Sao Paulo: análisis sobre razones de descarte corneal

Objetivo identificar os motivos da recusa de córneas. Método estudo transversal, retrospectivo, descritivo e correlacional, composto por 5.560 córneas ópticas. As informações foram extraídas do banco de dados da Central de Notificação, Captação e Doação de Órgãos (CNCDO), bem como de prontuários de doadores. A estatística descritiva foi utilizada para a análise das variáveis categóricas e testes específicos, com nível de significância de 5% para avaliar as associações entre as variáveis. Este estudo atendeu aos aspectos éticos da pesquisa científica. Resultados 60% dos doadores eram do sexo masculino e 40% morreram por problemas circulatórios. Os principais motivos de recusa informados pelas equipes de transplante são a idade do doador e a contagem de células endoteliais. Para cada ano adicionado à idade do doador, há uma redução de 1% na chance de que essa córnea seja usada para transplante, e para cada acréscimo de 100 células por mm2 aumenta as chances de que essa córnea seja usada em 9%. Conclusão a principal causa de recusa na aceitação do tecido corneano está relacionada à idade e à contagem de células endoteliais.

Objective to identify the reasons for refusal of corneas. Method this was a cross-sectional, retrospective, descriptive and correlational study composed of 5,560 optical corneas. The information was taken from the notification, organ procurement and distribution centers database as well as donor records. Descriptive statistics were used for the analysis of categorical variables and specific tests with a significance level of 5% for assessing the associations between variables. This study met the ethical aspects of scientific research. Results 60% of the donors were male and 40% died by circulatory problems. The main reason for refusal as informed by transplant teams is the donor's age and the endothelial cell count. For each year added to the donor's age, there is a 1% decrease in the chance that this cornea will be used for transplantation, and the increase of 100 cells per mm2 increases the chances that this cornea will be used by 9%. Conclusion the main cause of refusal in the acceptance of corneal tissue is related to the age and the endothelial cell count.

Objetivo identificar los motivos para el rechazo de córneas. Método estudio transversal, retrospectivo, descriptivo y correlacional, compuesto por 5.560 córneas ópticas. La información se obtuvo de la bases de datos de Centros de Notificación, Obtención de Órganos y Distribución, así como de los registros de los donantes. Se utilizaron estadísticas descriptivas para analizar las variables categóricas y pruebas específicas con un nivel de significación de 5%, para evaluar las asociaciones entre variables. Este estudio contempla los aspectos éticos de una investigación científica. Resultados el 60% de los donantes eran hombres y el 40% falleció por problemas circulatorios. La principal razón del rechazo, informada por los equipos de trasplante, es la edad del donante y el recuento de las células endoteliales. Por cada año agregado a la edad del donante, existe una disminución de 1% en la probabilidad de que la córnea pueda ser utilizada para trasplante; el aumento de 100 células por mm2 aumenta la posibilidad de que esta córnea será usada por 9%. Conclusión la principal causa de rechazo en la aceptación del tejido corneal está relacionada con la edad y el recuento de células endoteliales.

The Expression of Androgen Receptor and Its Variants in Human Prostate Cancer Tissue according to Disease Status, and Its Prognostic Significance

PURPOSE: To evaluate changes in the expression of androgen receptor (AR) and its variants (ARVs) in human prostate cancer (PCa) tissues according to disease status, and its prognostic significance following radical prostatectomy (RP). MATERIALS AND METHODS: A total of 282 PCa cases were evaluated, which included 252 localized PCa, 8 metastatic castration resistant prostate cancer (CRPC), and 22 benign prostatic hyperplasia (BPH) cases. Samples were collected from patients who underwent RP or transurethral resection and were stored in ethically approved tissue banks. Quantitative real-time polymerase chain reaction, Western blotting, and immunohistochemistry were performed for AR and ARVs. Each tissue was confirmed as cancerous (greater than 80%) using hematoxylin and eosin staining. AR and ARVs expression was compared according to disease status. The biochemical recurrence free survival (BCRFS) rates in men with localized PCa was analyzed according to AR and ARV7 expression using the Kaplan-Meier curve. RESULTS: Only 58 of the 252 localized PCa were included in the analysis because of insufficient cancer tissue. AR and ARV7 mRNA expression was higher in the CRPC tissue than in the localized PCa tissue (p=0.025, p=0.002, respectively). In localized PCa tissue, high AR mRNA and protein level was associated with a low BCRFS rate (log-ranked, p=0.019, p < 0.001, respectively). CONCLUSIONS: Overall AR and ARV7 mRNA expression levels were increased in CRPC tissues compared to localized PCa and BPH tissues. High AR protein and mRNA expression in the tumor tissue may be considered a predictive factor of BCRFS following RP.

Biobancos: os meus, os seus, os nossos / Biobanks: mine, yours, ours

O presente artigo tem como objetivo discutir aspectos conceituais, éticos e legais e também científicos e cooperativos dos acervos de materiais biológicos utilizados para pesquisa conhecidos como biobancos ou biorrepositórios humanos. Inicialmente são apresentados conceitos fundamentais sobre o tema, amplamente discutido internacional e nacionalmente e também aberta discussão sobre a presença destes acervos na Odontologia e nas Ciências Forenses, bem como sua situação em território nacional. O texto prossegue em três maiores blocos: O primeiro visa introduzir o leitor aos aspectos legais que cercam a temática dos biobancos e biorrepositórios, apresentando-o aos principais dispositivos que regulamentam o tema no Brasil. O segundo bloco aborda a importância do aspecto cooperativo e institucional que exercem os biobancos e biorrepositórios, expondo problemática existente acerca desse tema, mas evidenciando notoriamente os benefícios provenientes de uma postura receptiva dessas infraestruturas a pessoas externas. O terceiro e último bloco conecta o que foi anteriormente dito ao que é atualmente enfrentado cientificamente dentro da Odontologia Legal, referente à necessidade de uma maior variabilidade e número de amostras disponíveis ao pesquisador, que torne possível a este acompanhar e criar o dinamismo da evolução científica. Como conclusão, o artigo evidencia as vantagens e defende uma conduta receptiva a ser adotada pelos diferentes e variados centros e instituições de pesquisa que possuam biobancos ou biorrepositórios a pesquisadores como forma de elevar o nível científico produzido pela Odontologia Legal nacional.

The purpose of this article is to discuss conceptual, ethical and legal aspects, as well as scientific and cooperative aspects of the collections of biological materials used for research known as biobanks or human bio-repositories. Initially, fundamental concepts are presented on the subject, widely discussed internationally and nationally, and opens discussion about the presence of these collections within Dentistry and of Forensic Sciences, as well as their situation in national territory. The text aims to introduce the reader to the legal aspects that surround the biobanks and bio-repositories, presenting it to the main documents that regulate the subject in Brazil. The second section addresses the importance of the cooperative and institutional aspect of biobanks and bio-repositories, exposing existing problems on this topic, but highlighting the benefits of a receptive posture of these infrastructures to external people. The third and last section connects what was previously exposed to what is currently scientifically faced within Forensic Dentistry, referring to the need for a greater variability and number of samples available to the researcher, which makes it possible to follow up and create the dynamism of the scientific evolution. As a conclusion, the article highlights the advantages and advocates a receptive behavior to be adopted by different and varied research centers and institutions that have biobanks or bio-repositories to other researchers to raise the scientific level produced by national Forensic Dentistry.

Aspectos intervenientes na efetivação da doação do tecido ósseo durante o acolhimento e a entrevista familiar para doação de órgãos e tecidos / Aspects intervening in the effectiveness of bone tissue donation during the reception and the familiar interview to organs and tissues donation

Atualmente existe uma lacuna de conhecimento referente à doação e ao transplante do tecido ósseo, já que muitas publicações concentram-se, principalmente, na temática 'doação e transplante de órgãos sólidos'. O objetivo deste estudo foi compreender os aspectos intervenientes na efetivação da doação do tecido ósseo durante o acolhimento e a entrevista familiar, realizada pelos profissionais da Organização de Procura de Órgãos, na ótica do profissional e do familiar do potencial doador. Método: Este estudo de abordagem qualitativa, apoiado em conceitos do interacionismo simbólico, constituiu-se de familiares de doadores de órgãos e tecidos que autorizaram a doação do tecido ósseo no período de janeiro a dezembro de 2016, iniciando-se a partir de pesquisa nos prontuários da Organização de Procura de Órgãos localizada no Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto/SP. Selecionaram-se todos os prontuários dos doadores de tecido ósseo, totalizando doze doadores Ainda, foram convidados a participar do estudo os quatro enfermeiros responsáveis pelo acolhimento e entrevista para solicitação dos órgãos e tecidos pertencentes à Organização de Procura dos Órgãos citada, por meio de uma entrevista gravada e realizada em seu contexto de trabalho. Foram entrevistados nove familiares e quatro enfermeiros mediante roteiro de entrevista contendo dados sociodemográficos e questões sobre a vivência de todo o processo de doação do tecido ósseo. O material foi transcrito e submetido a análise de conteúdo indutiva em três categorias temáticas: contexto onde se dá o processo de solicitação da doação, fatores que facilitaram a decisão pela doação do tecido ósseo e o resultado do processo de doação para o familiar. Observamos que os familiares, ao refletirem sobre o momento de decisão pela doação do tecido ósseo, revelaram que, apesar do desconhecimento desse tipo de doação, os aspectos que contribuíram para o consentimento foram a satisfação com o atendimento hospitalar humanizado, bem como o profissionalismo dos enfermeiros da Organização de Procura de Órgãos ao explicarem quais ossos seriam retirados e como seria reconstituído o corpo do doador; o conhecimento prévio da vontade do ente querido em ser um doador e o pensamento de estar "fazendo o bem". Observamos ainda que os enfermeiros, ao refletirem sobre seu papel na entrevista para solicitação da doação do tecido ósseo, revelaram que, inicialmente, o desconhecimento dos familiares quanto à possibilidade de doação do tecido ósseo, faz com que os mesmos reajam negativamente devido ao medo da deformidade do corpo. Objetivando superar esse momento difícil, os enfermeiros explicam todo o processo de captação do tecido ósseo, até a forma como seria entregue o corpo do familiar para o velório. Sugerem-se capacitações futuras à equipe de enfermeiros contribuindo para a otimização de todo o processo de doação do tecido ósseo para transplante. Acreditamos ser evidente a necessidade de educação voltada à população por meio de ações governamentais, em que futuras campanhas na divulgação da existência da doação do tecido ósseo possam promover um maior entendimento, resultando em uma reação menos traumática das pessoas no momento da solicitação do tecido ósseo

Currently there is a lack of knowledge regarding donation and transplantation of bone tissue, due many publications to focus primarily on donation and transplantation of solid organs. The aim of this study was to understand the intervening aspects on the effectiveness of bone tissue donation during approach and family interview, carried out by professionals of the Organ Procurement Organization, from perspective of professionals and potential donor relatives. Methods: It is a qualitative study, based on symbolic interactionism concepts; the sample was composed for relatives of donors of organs and tissues that authorized the donation of bone tissue in the period from January to December of 2016,.The first part was characterized for a data collection in the medical records from Organ Procurement Organization located in a Brazilian School Hospital. There were twelve donors during the selected period which medical records were selected to analyze. In addition, four nurses belonging to Organ Procurement Organization were invited to participate in study. They were responsible for the approach and interview with relatives to request organs. The data collection strategy was a structured interview which was recorded and performed in their workplace. Nine families and four nurses were interviewed using an interview script containing sociodemographic data and questions about the experience of entire process of donating bone tissue. The interviews were transcribed and submitted to analysis of inductive content in three thematic categories: the context where the donation request process happens, factors that facilitated the decision by the donation of the bone tissue and the donation process result to the relative. Results: It was observed that family members reflected on the moment of decision of bone tissue donation. They revealed that, despite the lack of knowledge about this type of donation, there were some aspects that contributed to the consent satisfaction with humanized hospital care as well as the nurses of the Organ Procurement Organization professionalism. These nurses explained which bones would be removed and how the donor's body would be reconstituted. Another contributed factor was the relatives prior knowledge of the being´s desire to be an organ donor and the "doing good" feeling. Meanwhile, the nurses interviewed revealed that initially there was family members' lack of knowledge about the possibility of donation of the bone tissue, causes them to react negatively due to the fear of deformity of the body. The nurses explained the whole process of bone tissue withdrawal and the way which the deceased´s body would be delivered to the funeral. The aims of this approach were to overcome this difficult moment and family insecurity to make decision. Conclusions: Future nurses team training is suggested to improve all bone tissue donation for transplantation process. It is extremely important a population-oriented education through governmental actions, campaigns about bone tissue donation to promote a better understanding. It could result in less traumatic people reaction at the moment of bone tissue demand

Perfil epidemiológico do doador de pele do Banco de Tecidos do Hospital das Clínicas da Universidade de São Paulo / Epidemiological characteristics of skin donors from São Paulo University Clinical Hospital / El perfil epidemiológico de los donantes de piel del Banco de tejidos del Hospital de las Clínicas de la Universidad São Paulo

OBJETIVO: Identificar o perfil epidemiológico dos doadores de pele do Banco de Tecidos do Hospital das Clínicas da Universidade de São Paulo. MÉTODO: Trata-se de um estudo documental, retrospectivo, transversal, com abordagem quantitativa de análise de prontuários de doadores de pele do Banco de Tecidos do Hospital das Clínicas de São Paulo, no período entre 2000 a 2015, para identificar as principais características epidemiológicas dos doadores de pele sendo analisadas por método estatístico descritivo. Inicialmente, foram encontrados 183 prontuários, porém 15 foram excluídos devido à falta de informações completas dos doadores. Aprovado pela Comissão de Ética para Análise de Projetos de Pesquisa do HCFMUSP (Nº CAAE 42751915.5.0000.0068). RESULTADOS: Entre os doadores pesquisados, houve predomínio do sexo masculino, com média de idade 43,3 anos, sendo a principal causa dos óbitos os acidentes cerebrais vasculares e causas externas. A retirada do tecido ocorre em geral entre 1 e 4 horas (76,1% dos casos) após a parada cardíaca. A maior parte recebeu antibioticoterapia (92,2%) e não apresenta comorbidades (81,54%). CONCLUSÃO: O doador de pele em geral é do sexo masculino, jovem e previamente hígido, sendo fundamental esta caracterização para o fortalecimento dos mecanismos de busca de doadores e aumento da oferta de tecidos no país.(AU)

OBJECTIVE: To identify the skin donors epidemiological profile from Clinical Hospital Tissue Bank. METHODS: This is a retrospective, cross-sectional documentary study with a quantitative approach to analysis medical records of skin donors from Clínicas' Hospital Tissue Bank, between 2000 and 2015, to identify the main epidemiological characteristics of skin donors, being analyzed by descriptive statistical method. Initially, 183 medical records were found, but 15 were excluded due to lack of complete donor information. Approved by the Ethics Committee for Analysis of Research Projects of HCFMUSP (CAAE No. 42751915.5.0000.0068). RESULTS: Among the donors surveyed, there was a predominance of males, with a mean age of 43.3 years, the main cause of death was cerebrovascular accidents and external causes. Tissue withdrawal usually occurs between 1 and 4 hours (76.1% of cases) after cardiac arrest. The majority received antibiotic therapy (92.2%) and was healthy (81.54%). CONCLUSION: The skin donor in general is male, young and previously healthy, and this characterization is fundamental for the strengthening of donor search mechanisms and an increase in the supply of tissues in Brazil.(AU)

Objetivo: Identificar el perfil epidemiológico de los donantes del Banco de Tejidos del Hospital das Clínicas de la Universidad de São Paulo. Método: Se realizó un estudio documental, retrospectivo, transversal, con un enfoque cuantitativo para el análises de las histórias clínicas de los donadores de piel del Banco de tejidos del Hospital das Clínicas de São Paulo, en el periodo de 2000 a 2015, para identificar las principales características epidemiológicas de los donantes de piel, siendo analisados popr método estadístico descriptivo. Inicialmente, se encontraron 183 registros, pero 15 fueron excluidos debido a la falta de información completa de los donantes. Aprobado por el Comité de Ética en Investigación (núm. CAAE 42751915.5.0000.0068). Resultados: Entre los donantes estudiados, hubo un predominio del sexo masculino, una edad media de 43,3 años, es la principal causa de muerte de los accidentes vasculares cerebrales y las causas externas. El tejido extraído es generalmente entre 1 y 4 horas (76,1% de los casos) después de un paro cardíaco. La mayoría recibió antibióticos (92,2%) y muestra comorbilidades (81,54%). Conclusión: La piel del donante es generalmente de sexo masculino, jóvenes y previamente sanos, siendo fundamental la caracterización para fortalecer los mecanimos de bísqueda de donadores y aumento de la oferta de tejidos en el país.(AU)

Effects of ionizing radiation on proteins in lyophilized or frozen demineralized human bone / Efeitos da radiação ionizante nas proteínas presentes em ossos humanos desmineralizados, liofilizados ou congelados

OBJECTIVE: The aim was to study the effects of application of ionizing radiation (gamma and electrons) as sterilizing agents at doses of 15 kGy, 25 kGy and 50 kGy, on lyophilized or frozen demineralized bone tissue for use in transplants. METHODS: Five human femoral diaphyses from different donors of musculoskeletal tissue were demineralized and preserved as lyophilized or frozen at -80 °C. The samples were divided into two groups: non-irradiated (control) and irradiated by means of gamma rays or an electron beam. The bone proteins were extracted and used to determine the concentrations of total protein and BMP 2 and 7. RESULTS: Decreases in total protein and BMP 2 and 7 concentrations were observed. The decreases in total protein concentrations, in comparison with the respective control groups, were significant in the lyophilized and frozen samples that were irradiated at a dose of 50 kGy of gamma radiation and electron beam, with reductions of more than 30%. Significant decreases in the levels of BMP 2 and 7 were also observed at higher doses and especially through use of the electron beam. CONCLUSION: The reductions in the concentrations of total proteins and osteoinductive proteins (BMP 2 and 7) were related to the radiation dose, i.e. they increased with higher doses of ionizing radiation type and the type of bone preservation. The largest reductions in concentrations were observed in the bones irradiated by means of an electron beam and at a dose of 50 kGy. However, this type of radiation and this high dose are not usual practices for sterilization of bone tissue.

OBJETIVO: Estudar os efeitos da aplicação das radiações ionizantes (gama e elétrons) como agentes esterilizantes, nas doses de 15 kGy, 25 kGy e 50 kGy, nos tecidos ósseos desmineralizados congelados e liofilizados para uso em transplantes. MÉTODOS: Cinco diáfises femorais humanas de doadores distintos de tecidos musculoesqueléticos foram desmineralizadas e preservadas como liofilizadas ou congeladas a -80 °C. As amostras foram divididas em grupos não irradiados (controle) e irradiados por raios gama ou feixe de elétrons. As proteínas ósseas foram extraídas e dosadas as concentrações de proteínas totais, BMP 2 e 7. RESULTADOS: Foi observada diminuição das concentrações de proteínas totais e BMP 2 e 7. A diminuição das concentrações de proteínas totais, quando comparada com o respectivo controle, foi significativa nos grupos de amostras liofilizadas e congeladas e irradiadas na dose de 50 kGy por radiação gama e feixe de elétrons com redução superiores a 30%. A diminuição significativa nas concentrações das BMP 2 e 7 também foi observada nas maiores doses e principalmente por feixe de elétrons. CONCLUSÃO: As reduções nas concentrações das proteínas totais e em proteínas osteoindutoras (BMP 2 e 7) foram relacionadas à dose de radiação, ou seja, aumentam com maiores doses, tipo de radiação ionizante e ao tipo de preservação dos ossos. As maiores reduções das concentrações foram observadas nos ossos irradiados por feixe de elétrons e na dose de 50 kGy. Porém esse tipo de radiação e essa alta dose não são práticas usuais para a esterilização dos tecidos ósseos.

Doença por grãos argirofílicos / Argyrophilic grain disease

Introdução: A doença por grãos argirofílicos (DGA) é uma tauopatia esporádica distinta, bastante frequente, com uma prevalência atingindo 31,3% em centenários, porém pouco reconhecida A manifestação clínica mais comum da DGA é de um comprometimento cognitivo de lenta evolução associado a uma alta frequência de sintomas psiquiátricos. O diagnóstico de DGA é possível somente através da análise do encéfalo post-mortem com os achados das três principais alterações patológicas: grãos argirofílicos, corpúsculos em embrião e pré-emaranhados neuronais. O presente estudo investigou as características demográficas, clínicas e neuropatológicas dos indivíduos com DGA e possíveis associações clínico-patológicas. Métodos: Foram estudados 983 casos (acima de 50 anos de idade) provenientes da amostra do Banco de Encéfalos do Grupo de Estudos em Envelhecimento Cerebral. A avaliação clínica e funcional foi realizada através de uma ampla entrevista semiestruturada respondida por um informante com contato próximo com o paciente. Os participantes foram estratificados conforme a presença de comprometimento cognitivo (de acordo com Escala de Avaliação Clinica da Demência) e, posteriormente, pela presença de DGA em quatro grupos: DGA com e sem comprometimento cognitivo e não-DGA com e sem comprometimento cognitivo. Análise descritiva foi realizada para dados socioeconômicos, genótipo de APOE e variáveis clínico-funcionais, neuropsiquiátricas e neuropatológicas na amostra DGA e em cada grupo. Foi utilizado um modelo de regressão logística multivariada para investigar as associações entre perfil cognitivo e sintomas neuropsiquiátricos com DGA. Resultados: DGA foi identificada em 150 indivíduos (15,1%). Idade avançada e baixo nível socioeconômico foram associados com DGA independente da presença de comprometimento cognitivo. A presença de DGA foi associada a uma redução de 60% na probabilidade de um escore >= 3.8 no Questionário do informante sobre o Declínio Cognitivo do Idoso...

Background: Argyrophilic grain disease (AGD) is an underrecognized, distinct, highly frequent sporadic tauopathy, with prevalence reaching 31.3% in centenarians. The most common presentation of AGD is a slowly progressive amnestic mild cognitive impairment, accompanied by high frequency of neuropsychiatric symptoms. AGD can only be diagnosed postmortem by the finding of its three main pathologic features: argyrophilic grains, oligodendrocytic coiled bodies and neuronal pretangles. The present study investigated demographic, clinical, and neuropathological profiles and analyzed clinicopathological associations. Methods: We studied 983 participants (over 50 years of age) from the Brain Bank of the Brazilian Aging Brain study group sample. Clinical and functional evaluation included demographics and a semi-structured interview covering various cognitive domains conducted with a knowledgeable informant. Participants were stratified by cognitive status (based on Clinical Dementia Rating scale), followed by the presence of AGD in four groups: AGD with and without cognitive impairmet, and non-AGD with and without cognitive impairment. Descriptive statistics were used for sociodemographic data, APOE genotypes, and the clinical, cognitive, neuropsychiatric, functional, and neuropathological variables in AGD samples and in each group. We used multivariate logistic regression models to investigate the association between the cognitive status and neuropsychiatric symptoms with AGD. Results: AGD was identified in 150 participants (15.1%). Older age and lower socioeconomic status were associated with AGD independent of cognitive status. Multivariate analyses revealed that AGD was associated with a 60% reduction in the odds of having an IQCODE >= 3.8 (OR = 0.40, 95% CI 0.22-0.74, p = 0.004) and that the NPI sub-item "appetite and eating abnormalities" was associated with AGD in controls (OR = 1.85, 95% CI 1.09-3.12, p = 0.02). Conclusion: AGD...

Caracterização das alterações da proteína TDP-43 durante o envelhecimento normal: uma análise em cérebros humanos postmortem / Characterization of TDP-43 changes during normal aging: a human brain post-mortem study

INTRODUÇÃO: A proteína TDP-43 (Transactive DNA-binding protein 43) é o principal agregado proteico anormal verificado em casos de Degeneração Lobar Frontotemporal (DLFT) e Esclerose Lateral Amiotrófica (ELA). Apesar dos avanços na área, o papel da TDP-43 no envelhecimento normal ainda é pouco compreendido. A falta de indivíduos controles em estudos de associação clinicopatológica com tecido cerebral humano é resultado do alto custo da prática de autópsia mundialmente. O que é um diferencial do Brasil, onde essa prática é obrigatória em casos de morte natural sem causa definida. O Banco de Encéfalos Humanos do Grupo de Estudos em Envelhecimento Cerebral (BEHGEEC) possui grande número de encéfalos de sujeitos sem déficit cognitivo com alta miscigenação étnica e variação de graus de escolaridade. O que possibilita estudos mais aprofundados sobre envelhecimento cerebral humano não-patológico. OBJETIVOS: (1) investigar de maneira sistemática a distribuição dos achados neuropatológicos da TDP-43 no envelhecimento humano normal; (2) caracterizar a distribuição dessa proteína em diferentes áreas encefálicas; (3) explorar associação de características, clínicas, sociodemográficas ou neuropatológicas com o aparecimento das inclusões de TDP-43. MÉTODOS: Foram incluídos participantes com idade acima de 50 anos e classificados clinicamente e anatomopatologicamente como normais. Reação imunoistoquímica contra a conformação anormal da TDP-43 foi realizada em três regiões cerebrais de interesse córtex temporal, amígdala e hipocampo. RESULTADOS: Os agregados de TDP-43 estavam presentes em 10,5% dos indivíduos estudados (n=323). A região mais frequentemente acometida foi a amígdala (85,3% dos casos). O acúmulo de TDP-43 foi associado à idade e raça (p=0,002). Análise de regressão logística demonstrou que indivíduos da raça asiática possuem maior chance de apresentar os agregados de TDP-43 do que caucasianos, independente dos fatores gênero, idade, estágio de Braak para...

BACKGROUND: Transactive DNA-binding protein 43 (TDP-43) is the major abnormal aggregate present in Frontotemporal Lobar Degeneration (FTLD) and Amyotrophic Lateral Slcerosis (ALS). Although all the efforts in research in this field, the role played by TDP-43 in normal aging is still unknown. The lack of normal controls in studies focusing on clinicopathological associations is a result of the high cost of autopsy practice worldwide. In Brazil, autopsy is mandatory by law in cases without a certificate. The Brain Bank of the Brazilian Aging Study Group (BBBASG) comprises a large number of cognitively normal elderly subjects highly ethnically admixtured and with broad education attainment. This allow us to further study the non-pathological process of the human aging brain. GOALS: (1) sistematically investigate the distrubution of neuropathological findings of TDP-43 in the normal human brain; (2) characterize the distribution of theses findings in different brain regions; (3) explore clinical, sociodemographics or neuropathological variables that could be associated with TDP-43 inclusion outcome. METHODS: We included participants over 50 years old previously classified cognitively and neuropathologically as normals. Imunnohistochemistry against abnormal form of TDP-43 was performed in three brain regions: temporal cortex, hippocampal formation and amygdala. RESULTS: TDP-43 aggregates were present in 10,5% of the study subjects (n=323). Amygdala was the most frequently affected brain region (85.3% of the cases). TDP-43 accumulation was associated with age at death and race (p=0.002). Logistic regression analysis showed that asians older adults have higher odds of presenting TDP-43 inclusions than caucasians, regardless of gender, age, Braak stage for neurofibrilarly tangles and education attainment (OD=3.5, CI: 1.41-8.69, p=0.007). CONCLUSIONS: Our results suggest that TDP-43 abnormal accumulation increases along aging process and the amygdala is the brain...

Harnessing of corneas captured and processed for transplantation in an ocular tissue bank of North-eastern Brazil / Aprovechamiento de las córneas capturada y procesada para trasplante en un banco de tejidos oculares del Noreste / Aproveitamento das córneas captadas e processadas para transplante em um banco de tecidos oculares do Nordeste

The aim of this study was to describe the harnessing of corneas captured and processed for transplantation in an ocular tissue bank in north-eastern Brazil. This was a transverse and retrospective study, with a sample group of 612 individuals whose corneas were donated and captured between January/2007 and July/2012. This study was approved by the Research Ethics Committee under number 007.0.294.000-10, and research was based on an instrument consisting of social, demographic and clinical data of the donors. Of the 1209 corneas captured, 868 were used and 341 were discarded. Of the 612 donors, the corneas of 597 (97.5%) were captured from both eyes, being that 423 (70.9%) of these corneas were used. Further studies are required on the reasons for discarding corneas and clarifications as to the conduct of professionals when processing corneas, considering the increase in the quantity of donors and the elevated number of discarded ocular tissue.

El objetivo fue describir el aprovechamiento de las córneas capturada y procesada para trasplante en un banco de tejidos oculares del noreste. Estudio transversal y retrospectivo, con una muestra de 612 personas que tenían sus córneas donadas y capturados entre enero/2007 a julio/2012. Después de la aprobación del Comité de Ética de Investigación de número 007.0.294.000-10, se utilizó un instrumento que consiste en datos sociodemográficos y clínicos de los donantes. De las 1.209 córneas capturadas, 868 se utilizaron y 341 se desecharon. De los 612 donantes, 597 (97,5%) tenían capitación de las córneas de los dos ojos, entre estos, se utilizaron 423 (70,9%). Necesidad de ser de más estudios sobre las causas de los vertidos de la córnea y aclaraciones a la conducta de los profesionales en el procesamiento de estos, al ver el aumento en la cantidad de donantes y el elevado número de desechados de los tejidos oculares.

Objetivou-se descrever o aproveitamento das córneas captadas e processadas para transplante em um banco de tecidos oculares do Nordeste. Estudo transversal e retrospectivo, com amostra de 612 pessoas que tiveram suas córneas doadas e captadas, entre janeiro de 2007 a julho de 2012. Após a aprovação pelo Comitê de Ética em Pesquisa, de número 007.0.294.000-10, utilizou-se um instrumento composto por dados sociodemográficos e clínicos dos doadores. Das 1.209 córneas captadas, 868 foram utilizadas e 341 descartadas. Dos 612 doadores, 597 (97,5%) tiveram captação das córneas dos dois olhos e, dentre estas, 423 (70,9%) foram utilizadas. Necessita-se de mais estudos sobre causas de descartes de córneas e esclarecimentos quanto à conduta dos profissionais no processamento destas, visto o aumento da quantidade de doadores e o número elevado de descartes de tecidos oculares.

Fatores envolvidos na negativa da doação de tecido ósseo / Factors involved in the refusal to donate bone tissue

Objetivo: Determinar as principais variáveis envolvidas na negativa da doação de tecido ósseo em familiares doadores de órgãos e a prevalência da doação desse tecido. Métodos: Trata-se de um estudo transversal, quantitativo. Foram estudadas variáveis referentes à solicitação da doação do tecido ósseo, compreendendo informações a respeito dos ossos que seriam captados, a reconstituição do corpo, os motivos que levaram à negativa da doação desse tecido, conhecimentos anteriores sobre doação e intencionalidade em ser doador. Resultados: Evidenciamos três fatores que influenciaram a negativa pelos familiares incluindo o desconhecimento sobre quais ossos seriam retirados (92,9%), como ocorreria a reconstituição destas estruturas (96,5%) e como se apresentaria o corpo após a captação. A prevalência de doadores de tecido ósseo foi de 17,2%. Conclusão: A decisão familiar de recusar a doação do tecido ósseo foi representada pelo desconhecimento do tema. A baixa prevalência foi devido à reduzida solicitação deste tipo de doação. .

Objective: To determine the main variables involved in the refusal to donate bone tissue among relatives of organ donors, and the prevalence of bone tissue donation. Methods: This was a cross-sectional, quantitative study. Variables related to donation requests for bone tissue were studied, comprising information about the bones that would be procured, the reconstruction of the body, the reasons that led to the refusal to donate bone tissue, prior knowledge about donation, and intention to become a donor. Results: We demonstrated three factors that influenced the refusal of family members, including: lack of understanding of which bones would be removed (92.9%), how the body would be reconstructed after bone removal (96.5%), and how the body would be presented after bone procurement. The prevalence of bone tissue donation was 17.2%. Conclusion: The family decision to refuse to donate bone tissue was represented by the lack of comprehension of the subject. The low prevalence was due to the reduced request of this type of donation. .

An outcome analysis and long-term viability of cryopreserved cultured epidermal allografts: assessment of the conservation of transplantable human skin allografts

PURPOSE: To assess the viability of cultured epithelium and preserved by freezing for periods varying from one month to one year. METHODS: Samples of cultured epithelium were incubated in cryoprotectant medium (Group A), packed in aluminum envelopes and packed in polystyrene boxes. The boxes were subjected to a temperature of-70ºC. After freezing for a period of time ranging from one to 12 months, cultured epithelial samples were assessed for their viability by vital staining (Trypan blue) and metabolic analysis based on glucose consumption and lactate production. Samples of not frozen cultured epithelium (Group B) were also tested for viability and the results obtained were used as comparison parameter for the variation of viability. RESULTS: Statistical analysis between the group A and B indicate that the mean age of the donors (p=0.51) and the culture time (p=1.18) showed no statistical difference. In 30 days we obtained 37% of the original viability of cultured epithelium, 25% at six months and one year, less than 15%. This trend was confirmed statistically with a reduction of approximately 1.8% of the original viability epithelium cultured every 30 days of storage. In the analysis by lactate production, similar results were observed. In the analysis by the glucose consumption results were not significant. The viability indices show statistically significant difference between the group A and B (p<0.0001). CONCLUSIONS: Although cryopreserved cultured epithelium showed significant reduction of viability, all samples remained viable. It was also found that the viability of cryopreserved cultured epithelial decreased as a function of storage time.

Adoption of Donor Screening Policy in a Tissue Bank at a Tertiary Hospital / 대한수혈학회지

BACKGROUND: Tissues for transplantation can save lives or restore essential functions. According to national policies and regulations, access to suitable transplantation, as well as the level of safety, quality, efficacy of donation, and transplantation of tissues, differ significantly between countries. We reviewed a few guidelines on tissue banking from the aspect of screening tests. In addition, four-year experience with screening panels for donated bones and donors at a tertiary hospital is introduced. METHODS: Seven national and international guidelines for screening tests for donors and donated tissues were reviewed. At our institution, screening tests for donation involve two steps. At retrieval, the first screening panel, including ABO/Rh typing, unexpected antibody screening, VDRL, HBsAg, anti-HBs, anti-HBc IgM, anti-HCV, anti-HIV, and microbiological cultures was performed. The second screening panel, including the same tests, except culture studies, was performed after 90 days. From 2008 to 2011, a total of 245 retrievals of bone tissue were performed and the screening panel results were analyzed. RESULTS: Mandatory screening serologic tests for living donors can differ according to local law or regulation and/or screening for endemic diseases. At our institution, among 245 donated bones for a period of four years, 61 bone tissues were discarded due to noncompliance for the second screening (n=32), contamination or no culture study results (n=9), abnormal serologic test results (n=8), and so on. CONCLUSION: Donor screening policies for tissue banking are various according to national laws or endemic disease status. Second screening tests with consideration of the window period should be adopted.

Homoenxertos ósseos: a captação e o processamento geram segurança para a utilização? / Bone allografts: Does the collecting and processing method generate security for its use?

Frente à popularização das reabilitações bucais com próteses sobre implantes e da ausência, por vezes, de suficiente ou adequado tecido ósseo, a demanda dos enxertos ósseos tem aumentado progressivamente. Nas situações de extenso déficit ósseo ou frente à necessidade de reduzir o tempo e a morbidade cirúrgica, o uso de homoenxertos ósseos deve ser considerado, já que são encontrados em quantidades ilimitadas nos bancos de tecidos. No entanto, a possibilidade da transmissão de doenças infecciosas é uma constante preocupação dos profissionais e dos pacientes envolvidos. O presente trabalho tem como objetivo, por intermédio de uma revisão de literatura — realizada manualmente e através da base de dados Medline, compreendendo o período de 1990 a 2010 —, verificar a segurança oferecida pela captação e pelo processamento dos homoenxertos ósseos. Pode-se concluir que as etapas do processamento como a liofilização, a desmineralização e a irradiação gama geram níveis de segurança confiáveis e que, associados a uma prévia seleção de doadores e captação dos homoenxertos criteriosas, reduzem a possibilidade de infecção, assim gerando segurança.

The popularization of oral rehabilitation with prostheses on implants increased the demand of bone grafts. The use of bone allografts should be considered in situations of extensive bone deficit or when facing the reduction of time and surgical morbidity, since they are available in unlimited amounts in tissue banks. However, the possibility of infectious diseases' transmission is a common concern by the professionals and the involved patients. This paper aims to verify the security of collecting and processing bone allografts through a literature review that was performed manually covering Medline database from 1990 to 2010. It can be concluded that the processing steps such as freeze drying, demineralization and gamma radiation are reliable and in association with a judicious donor selection and collection of bone allograft performed previously, reduce the chance of infection, thereby creating safety.

Tumor banks: the cornerstone of basic research in urology

PURPOSE: Tumor banks have the primary responsibility for collecting, cataloging, storing and disseminating samples of tissues, cells and fluids, which are used by researchers to identify diagnostic molecular markers, prognostic indicators and therapeutic targets. The objective of this review was to describe a simple, reliable and reproducible protocol for obtaining and storing samples of urological tumors. MATERIALS AND METHODS: Urogenital tumor tissues were collected by the surgeons from the Urology Division of University of Sao Paulo Medical School. The obtained surgical specimens were immediately placed in liquid nitrogen, dry ice or in a tube containing RNAlater ®, and then stored by cryopreservation (-80°C). A mirror fragment was fixed in 10 percent formalin processed routinely and embedded in Paraplast®. RESULTS: We developed a protocol for the collection, cataloging, storage, conservation and use of tumor samples. During a period of one year the Urological Tumor Bank of the Urology Division stored 274 samples of prostate, bladder, kidney, penis and testicle tumors of different histological types, 74 urine and 271 serum samples. CONCLUSIONS: Having biological materials characterized and available along with the clinical patient information provides an integrated portrait of the patients and their diseases facilitating advances in molecular biology. It also promotes the development of translational research improving methods of diagnosis and cancer treatment.

Advances in standardized operational processes of tumor tissue bank / 国际肿瘤学杂志

The standard operating procedures (SOP) of tumor tissue bank (TTB) involves the collection, processing, storage, quality control of biological samples. It is also the key to the normal operation of TTB and strengthen the cooperation between them. Many countries have recognized the need for the establishment of TTB, but the prevailing issue is that TTB is lack of access to a set of SOP for high-quality tissue samples. To further standardize the construction of TTB will also be involved in information management and common data elements (CDE) development of TTB.

Evaluation of vitrification for cryopreservation of teeth

PURPOSE: The aim of this study was to investigate whether vitrification in the cryopreservation of periodontal ligament (PDL) cells could be useful for tooth banking. METHODS: In step 1, primary cultured human PDL cells were cryopreserved in 100% conventional cryopreservation media and 100% vitrification media (ESF40 media) in different temperatures for 2 weeks. In step 2, a series of modified vitrification formulae named T1 (75% vitrification media + 25% F media), T2 (50% vitrification media + 50% F media) and T3 (25% vitrification media + 75% F media) were used to store PDL cells for 2 weeks and 4 weeks in liquid nitrogen. MTT assay was performed to examine the viability of PDL cells. RESULTS: Maximum cell viability was achieved in cells stored in 100% conventional cryopreservation media at -196degrees C (positive control group) in step 1. Compared to the positive control group, viability of the cells stored in 100% vitrification media was very low as 10% in all test conditions. In step 2, as the percentage of vitrification media decreased, the cell viability increased in cells stored for 2 weeks. In 4-week storage of cells in step 2, higher cell viability was observed in the T2 group than the other vitrification formulae while the positive control group had the highest viability. There was no statistically significant difference in the cell viability of 2-week and 4-week stored cells in the T2 group. CONCLUSIONS: These observations indicate 100% vitrification media is not successful in PDL cell cryopreservation. Conventional cryopreservation media is currently the most appropriate media type for this purpose while T2 media would be interesting to test for long-term storage of PDL cells.