Tissue banks are thought to be an essential resource for medical research in the post-genomic age. Collections of tissue, usually removed in the course of diagnostic or therapeutic procedures, enable laboratory-based epidemiological studies to be carried out, linking abnormalities in the tissue to disease aetiology, prognosis and treatment responsiveness. There are, however, a number of technical, regulatory and ethical concerns that challenge those wishing to engage in tissue banking research. It is becoming increasingly apparent that tissue banking research is not without risk of harms, even though there is no direct physical risk to donors. This is because, in order to be most useful, banked specimens need to be linked to personal information about tissue donors and this poses the risk of inadvertent disclosure of personal─ particularly genetic─ information to those who might exploit such information (eg. insurance companies and employers). Furthermore, the long-term storage of specimens, and the impossibility of predicting all potential types of research programs for which they might be useful, raises the possibility that future projects will be carried out that are unacceptable to some (past) tissue donors. The ethical principles of autonomy and respect for persons demand that research subjects be informed of such risks and of the nature of the research, and that they participate willingly. On the other hand, there is a desire for science to progress unhindered by stringent consent requirements. For these reasons, a debate has emerged in the academic (bioethical and biomedical) literature and in the legal (law reform) sphere over what would constitute adequate consent. Despite an extensive discourse, it is still unclear whether it is permissible to carry out research on archival tissue that was originally taken for diagnostic purposes and whether project-specific (as opposed to open-ended) consent is required for research on tissue collected today. This lack of clarity is of concern to researchers, ethics committees and research subjects, all of whom recognise the importance of tissue banking research, yet fear that current consent procedures may be ethically or legally inadequate. Thus it is important that the consent dilemma be resolved as quickly and definitively as possible. Ongoing controversy and regulatory ambiguity are appropriate when morally contentious issues are at stake, and their existence does not, on its own, signal any flaws in the discourse process. There are, however, two reasons to suspect that the current �consent to tissue banking� debate, as portrayed in the academic literature and law reform documentation, is problematic. Firstly, the debate appears to be mired in an intractable conflict between those who want to maximise personal autonomy through stringent consent requirements, and those who want the scientific endeavour to progress in a manner that is unconstrained by what are viewed as arduous consent procedures. Secondly, the possible practical options (consent models) being generated by the debate are all limited because they are underpinned by a restricted notion of consent as an individualistic, legalistic and static activity, without consideration of any alternative conceptualisations of consent. Through a thematic analysis of the current �consent to tissue banking� debate in the academic and law reform literature (Section 3), this thesis shows that debate is essentially occurring between those who see individual autonomy (and stringent consent) as being of primary importance, and those who see unimpeded, market-driven scientific progress as the more important social good, which should not be impeded by unnecessarily stringent consent. Thematic analysis also confirms the existence of the two problems described above, and a failure of those engaged in the debate to reflect on, and challenge, the value-level assumptions underpinning their arguments and those of their opponents. It is argued that this lack of reflection accounts for the two problems: � Firstly, it precludes recognition of the cause of─ and, therefore, ways of resolving─ the intractable conflict at the centre of the debate. Value-level reflection shows that this is a result of the logical and moral conflict within western liberalism, between two modernist goods: individual freedom and scientific progress. � Secondly, it precludes the generation of varied conceptions of consent. Value-level reflection shows that the current range of consent models is restricted to procedures which are individualistic, abstract, static and legalistic, since they are underpinned by western liberal notions of autonomy and scientific progress. This recognition paves the way to consideration of alternative notions of autonomy, scientific progress and, therefore, consent, such as those derived from communitarian and feminist systems of values. A conceptually enriched model of tissue banking consent is then developed (Section 4). This model incorporates dominant (liberal) conceptions of autonomy and scientific progress as well as alternative notions of autonomy and scientific progress espoused by communitarian and feminist systems of values. It is argued that this conceptually-enriched model provides a practical solution to the two problems associated with the standard �consent to tissue banking� debate. In relation to the philosophically intractable conflict─ or what is termed the �modernist dilemma�─ between those privileging autonomy and those privileging scientific progress, it shows how the two apparently conflicting �modernist� goods can both be accommodated at a practical level, thus making the �consent to tissue banking� debate more tractable and fruitful. In relation to the restricted range of consent models being generated by the current debate, it provides new insights into the ways in which consent might be obtained such that a broader range of community values can be accommodated. More specifically, it stimulates the construction of a model that 1) involves communities, as opposed to merely individuals, in all stages of the scientific process; 2) is flexible and able to adapt consent procedures to specific contexts, rather than predefining procedures in abstract terms; and 3) is transactional and relational rather than static and legalistic. This outcome has interesting philosophical as well as practical implications. It shows that despite apparently unresolved, and possibly irresolvable, normative-level conflicts between the two modernist elements of western liberalism (autonomy and scientific progress), and between liberal, feminist and communitarian systems of values, a multi-perspectival, inclusive, model-building approach provides a practical solution that circumvents these normative-level conflicts.

Tissue banks are thought to be an essential resource for medical research in the post-genomic age. Collections of tissue, usually removed in the course of diagnostic or therapeutic procedures, enable laboratory-based epidemiological studies to be carried out, linking abnormalities in the tissue to disease aetiology, prognosis and treatment responsiveness. There are, however, a number of technical, regulatory and ethical concerns that challenge those wishing to engage in tissue banking research. It is becoming increasingly apparent that tissue banking research is not without risk of harms, even though there is no direct physical risk to donors. This is because, in order to be most useful, banked specimens need to be linked to personal information about tissue donors and this poses the risk of inadvertent disclosure of personal─ particularly genetic─ information to those who might exploit such information (eg. insurance companies and employers). Furthermore, the long-term storage of specimens, and the impossibility of predicting all potential types of research programs for which they might be useful, raises the possibility that future projects will be carried out that are unacceptable to some (past) tissue donors. The ethical principles of autonomy and respect for persons demand that research subjects be informed of such risks and of the nature of the research, and that they participate willingly. On the other hand, there is a desire for science to progress unhindered by stringent consent requirements. For these reasons, a debate has emerged in the academic (bioethical and biomedical) literature and in the legal (law reform) sphere over what would constitute adequate consent. Despite an extensive discourse, it is still unclear whether it is permissible to carry out research on archival tissue that was originally taken for diagnostic purposes and whether project-specific (as opposed to open-ended) consent is required for research on tissue collected today. This lack of clarity is of concern to researchers, ethics committees and research subjects, all of whom recognise the importance of tissue banking research, yet fear that current consent procedures may be ethically or legally inadequate. Thus it is important that the consent dilemma be resolved as quickly and definitively as possible. Ongoing controversy and regulatory ambiguity are appropriate when morally contentious issues are at stake, and their existence does not, on its own, signal any flaws in the discourse process. There are, however, two reasons to suspect that the current �consent to tissue banking� debate, as portrayed in the academic literature and law reform documentation, is problematic. Firstly, the debate appears to be mired in an intractable conflict between those who want to maximise personal autonomy through stringent consent requirements, and those who want the scientific endeavour to progress in a manner that is unconstrained by what are viewed as arduous consent procedures. Secondly, the possible practical options (consent models) being generated by the debate are all limited because they are underpinned by a restricted notion of consent as an individualistic, legalistic and static activity, without consideration of any alternative conceptualisations of consent. Through a thematic analysis of the current �consent to tissue banking� debate in the academic and law reform literature (Section 3), this thesis shows that debate is essentially occurring between those who see individual autonomy (and stringent consent) as being of primary importance, and those who see unimpeded, market-driven scientific progress as the more important social good, which should not be impeded by unnecessarily stringent consent. Thematic analysis also confirms the existence of the two problems described above, and a failure of those engaged in the debate to reflect on, and challenge, the value-level assumptions underpinning their arguments and those of their opponents. It is argued that this lack of reflection accounts for the two problems: � Firstly, it precludes recognition of the cause of─ and, therefore, ways of resolving─ the intractable conflict at the centre of the debate. Value-level reflection shows that this is a result of the logical and moral conflict within western liberalism, between two modernist goods: individual freedom and scientific progress. � Secondly, it precludes the generation of varied conceptions of consent. Value-level reflection shows that the current range of consent models is restricted to procedures which are individualistic, abstract, static and legalistic, since they are underpinned by western liberal notions of autonomy and scientific progress. This recognition paves the way to consideration of alternative notions of autonomy, scientific progress and, therefore, consent, such as those derived from communitarian and feminist systems of values. A conceptually enriched model of tissue banking consent is then developed (Section 4). This model incorporates dominant (liberal) conceptions of autonomy and scientific progress as well as alternative notions of autonomy and scientific progress espoused by communitarian and feminist systems of values. It is argued that this conceptually-enriched model provides a practical solution to the two problems associated with the standard �consent to tissue banking� debate. In relation to the philosophically intractable conflict─ or what is termed the �modernist dilemma�─ between those privileging autonomy and those privileging scientific progress, it shows how the two apparently conflicting �modernist� goods can both be accommodated at a practical level, thus making the �consent to tissue banking� debate more tractable and fruitful. In relation to the restricted range of consent models being generated by the current debate, it provides new insights into the ways in which consent might be obtained such that a broader range of community values can be accommodated. More specifically, it stimulates the construction of a model that 1) involves communities, as opposed to merely individuals, in all stages of the scientific process; 2) is flexible and able to adapt consent procedures to specific contexts, rather than predefining procedures in abstract terms; and 3) is transactional and relational rather than static and legalistic. This outcome has interesting philosophical as well as practical implications. It shows that despite apparently unresolved, and possibly irresolvable, normative-level conflicts between the two modernist elements of western liberalism (autonomy and scientific progress), and between liberal, feminist and communitarian systems of values, a multi-perspectival, inclusive, model-building approach provides a practical solution that circumvents these normative-level conflicts.

O crescente aumento do interesse no desenvolvimento em técnicas cirúrgicas menos invasivas, como nas reconstruções tendíneas e ligamentares, tem levado ao aumento das pesquisas referentes ao uso de aloenxertos esterilizados por radiação ionizante. O processamento por radiação ionizante é um método seguro e não deixa resíduos, sendo utilizado como esterilização final. O presente estudo teve como proposta avaliar os efeitos da aplicação de radiação ionizante, produzida por fonte de 60Co, em amostras de tendões humanos pré processados de doadores multiorgãos obtidas por meio de colaboração com Bancos de Tecidos. O pré-processamento das amostras deu-se por métodos químicos e preservação por congelamento em -80 °C. As doses aplicadas no processamento por radiação foram de 12,5 kGy, 15,0 kGy e 25,0 kGy, cada uma com seu respectivo controle não irradiado. As amostras foram avaliadas por meio de testes histológicos, ópticos e biomecânicos, com o objetivo de analisar possíveis modificações morfológicas e estruturais. Os resultados apresentados demonstraram que o processamento por ultrassom e peróxido de hidrogênio causaram alterações na morfologia dos tecidos, o que ocasionou danos à sua estrutura, inviabilizando as amostras. Nas amostras processadas por álcool e antibiótico não foram observados danos na rede de colágeno pela aplicação da radiação. Os resultados dos testes biomecânicos apresentaram diferenças significativas entre os métodos aplicados. As amostras processadas com álcool e antibiótico apresentaram perda pouco significativa no módulo de elasticidade, comparadas às amostras processadas por ultrassom e peróxido de hidrogênio que mantiveram a propriedade viscoelástica. Contudo na dose de 12,5 kGy foi observado um aumento no módulo elástico e na viscoelasticidade. Com base nas análises, podemos concluir que o método de processamento com álcool, antibiótico e irradiação demonstrou menor dano, tanto na biomecânica quanto na esterilização, sendo que as amostras irradiadas a 15,0 e 25,0 kGy apresentaram características semelhantes ao controle não irradiado. / The increasing interest of development in less invasive surgical techniques, such as reconstructions of ligament tendon, has led to the increase of the research concerning the use of Allografts sterilized by ionizing radiation. Processing by ionizing radiation is a safe method and leaves no residues, being used as final sterilization. The present study was to evaluate the effects of proposed application of ionizing radiation, produced by 60Co source in human tendon pre-samples processed multiorgans donors obtained through collaboration with tissue banks. The pre-processing of samples given by chemical methods and preserved by freezing at -80 °C. The doses applied in radiation processing were 12.5, 15.0 and 25.0 kGy, each with your corresponding non-irradiated control. The samples were evaluated by means of histological and biomechanical testing, with the purpose of analyzing possible structural and morphological changes. The results showed that the ultrasound processing and hydrogen peroxide caused changes in the morphology of the tissues, which caused damage to the structure, making your samples. In the samples processed by alcohol and antibiotics were not observed damage on the network of collagen by the application of radiation. The results of biomechanical tests showed significant differences between the methods used. The samples processed with alcohol and antibiotics showed negligible loss in modulus of elasticity compared with the samples processed by ultrasound and hydrogen peroxide which kept the viscoelastic property, however in 12.5 kGy dose was observed an increase in elastic modulus and viscoelasticity. Based on the analysis we can conclude that the method of processing with alcohol, antibiotics and irradiation showed less damage, both in biomechanics and sterilization, in the samples irradiated with 15.0 and 25.0 kGy, showing results similar to the non-irradiated control.

Diversos pacientes são beneficiados com ossos armazenados em Bancos de Tecidos e utilizados em cirurgias reconstrutivas ortopédicas e em implantodologia como aloenxertos. No entanto, há uma intensa preocupação em garantir segurança na esterilidade do aloenxerto para proporcionar eficácia no transplante. Para minimizar possíveis contaminações utiliza-se a radiação ionizante como forma de esterilização final, desde que esta seja feita de maneira controlada, evitando possíveis modificações na matriz óssea. No presente trabalho, utilizamos as técnicas de colorimetria para avaliar modificações estéticas, Tomografia por Coerência Óptica, Tomografia por Coerência Óptica sensível à polarização, espectroscopia Raman e ensaios mecânicos de compressão para identificar as possíveis alterações na matriz óssea, ocasionadas pela forma de preservação, assim como, pelas diferentes doses de irradiação. Foram obtidas 8 amostras de fíbulas de 4 doadores, fracionadas de maneira a obter 48 amostras liofilizadas e 48 amostras congeladas. As amostras foram irradiadas com as doses de 15, 25 e 50 kGy comparando os resultados com o controle não irradiado. Observamos uma diminuição na intensidade das cores iniciais, mais relacionada com o processamento e preservação das amostras, e para as amostras irradiadas somente foi observado um aumento da coloração amarelada na dose de 50 kGy. A forma de preservação por liofilização ocasionou maiores modificações na estrutura terciária do colágeno dos ossos irradiados nas diferentes doses, principalmente nas doses acima de 25 kGy, porém estas modificações não foram suficientes para alterar a organização das fibras de colágeno. Quanto à resistência mecânica, verificou-se que as amostras liofilizadas foram menos resistentes que as congeladas e nas doses de 15 kGy e 25 kGy em ambas as formas de preservação ocorreu uma tendência a diminuir a resistência mecânica em relação ao controle. / Several patients are benefited with bones stored in Tissue Banks and used in orthopedic reconstructive surgery and implantodology as allografts. However, there is a strong concern to ensure safety in sterile allograft transplantation in order to provide efficacy. To minimize a probable contamination, ionizing radiation is used as a form of final sterilization, since the procedure is done in a controlled manner, avoiding possible changes in the bone matrix. In this dissertation, the techniques of colorimetry were used to evaluate aesthetic modifications; Optical Coherence Tomography, Optical Coherence Tomography sensitive to polarization, Raman spectroscopy and mechanical compression was carried out to identify possible changes in the bone matrix, caused by the preservation method, as well as the different irradiation doses. Eight fibulae from four donors were fractionated and from that forty-eight lyophilized samples and forty-eight frozen samples were obtained. The samples were irradiated with doses of 15, 25 and 50 kGy and the results were compared with the non-irradiated control. A decrease in the intensity of the initial colors was noticed and it was more related to the processing and preservation of the samples; for the irradiated samples, only an increase in the yellowness was observed, in the 50 kGy doses. The lyophilization preservation method caused major changes in the tertiary structure of the bone collagen irradiated at different doses, particularly at doses above 25 kGy, but these changes were not enough to change the organization of collagen fibers. Regarding the mechanical strength, were detected that lyophilized samples were less resistant than those which were frozen. With doses of 25 kGy and 15 kGy, in both forms of preservation, the mechanical strength tended to decrease, compared to the control. Therefore, it was observed that the aesthetic and structural changes caused by the different irradiation doses depend on the processing used in the Tissue Banks, besides the preservation method selected.

Diversos pacientes são beneficiados com ossos armazenados em Bancos de Tecidos e utilizados em cirurgias reconstrutivas ortopédicas e em implantodologia como aloenxertos. No entanto, há uma intensa preocupação em garantir segurança na esterilidade do aloenxerto para proporcionar eficácia no transplante. Para minimizar possíveis contaminações utiliza-se a radiação ionizante como forma de esterilização final, desde que esta seja feita de maneira controlada, evitando possíveis modificações na matriz óssea. No presente trabalho, utilizamos as técnicas de colorimetria para avaliar modificações estéticas, Tomografia por Coerência Óptica, Tomografia por Coerência Óptica sensível à polarização, espectroscopia Raman e ensaios mecânicos de compressão para identificar as possíveis alterações na matriz óssea, ocasionadas pela forma de preservação, assim como, pelas diferentes doses de irradiação. Foram obtidas 8 amostras de fíbulas de 4 doadores, fracionadas de maneira a obter 48 amostras liofilizadas e 48 amostras congeladas. As amostras foram irradiadas com as doses de 15, 25 e 50 kGy comparando os resultados com o controle não irradiado. Observamos uma diminuição na intensidade das cores iniciais, mais relacionada com o processamento e preservação das amostras, e para as amostras irradiadas somente foi observado um aumento da coloração amarelada na dose de 50 kGy. A forma de preservação por liofilização ocasionou maiores modificações na estrutura terciária do colágeno dos ossos irradiados nas diferentes doses, principalmente nas doses acima de 25 kGy, porém estas modificações não foram suficientes para alterar a organização das fibras de colágeno. Quanto à resistência mecânica, verificou-se que as amostras liofilizadas foram menos resistentes que as congeladas e nas doses de 15 kGy e 25 kGy em ambas as formas de preservação ocorreu uma tendência a diminuir a resistência mecânica em relação ao controle. / Several patients are benefited with bones stored in Tissue Banks and used in orthopedic reconstructive surgery and implantodology as allografts. However, there is a strong concern to ensure safety in sterile allograft transplantation in order to provide efficacy. To minimize a probable contamination, ionizing radiation is used as a form of final sterilization, since the procedure is done in a controlled manner, avoiding possible changes in the bone matrix. In this dissertation, the techniques of colorimetry were used to evaluate aesthetic modifications; Optical Coherence Tomography, Optical Coherence Tomography sensitive to polarization, Raman spectroscopy and mechanical compression was carried out to identify possible changes in the bone matrix, caused by the preservation method, as well as the different irradiation doses. Eight fibulae from four donors were fractionated and from that forty-eight lyophilized samples and forty-eight frozen samples were obtained. The samples were irradiated with doses of 15, 25 and 50 kGy and the results were compared with the non-irradiated control. A decrease in the intensity of the initial colors was noticed and it was more related to the processing and preservation of the samples; for the irradiated samples, only an increase in the yellowness was observed, in the 50 kGy doses. The lyophilization preservation method caused major changes in the tertiary structure of the bone collagen irradiated at different doses, particularly at doses above 25 kGy, but these changes were not enough to change the organization of collagen fibers. Regarding the mechanical strength, were detected that lyophilized samples were less resistant than those which were frozen. With doses of 25 kGy and 15 kGy, in both forms of preservation, the mechanical strength tended to decrease, compared to the control. Therefore, it was observed that the aesthetic and structural changes caused by the different irradiation doses depend on the processing used in the Tissue Banks, besides the preservation method selected.

O crescente aumento do interesse no desenvolvimento em técnicas cirúrgicas menos invasivas, como nas reconstruções tendíneas e ligamentares, tem levado ao aumento das pesquisas referentes ao uso de aloenxertos esterilizados por radiação ionizante. O processamento por radiação ionizante é um método seguro e não deixa resíduos, sendo utilizado como esterilização final. O presente estudo teve como proposta avaliar os efeitos da aplicação de radiação ionizante, produzida por fonte de 60Co, em amostras de tendões humanos pré processados de doadores multiorgãos obtidas por meio de colaboração com Bancos de Tecidos. O pré-processamento das amostras deu-se por métodos químicos e preservação por congelamento em -80 °C. As doses aplicadas no processamento por radiação foram de 12,5 kGy, 15,0 kGy e 25,0 kGy, cada uma com seu respectivo controle não irradiado. As amostras foram avaliadas por meio de testes histológicos, ópticos e biomecânicos, com o objetivo de analisar possíveis modificações morfológicas e estruturais. Os resultados apresentados demonstraram que o processamento por ultrassom e peróxido de hidrogênio causaram alterações na morfologia dos tecidos, o que ocasionou danos à sua estrutura, inviabilizando as amostras. Nas amostras processadas por álcool e antibiótico não foram observados danos na rede de colágeno pela aplicação da radiação. Os resultados dos testes biomecânicos apresentaram diferenças significativas entre os métodos aplicados. As amostras processadas com álcool e antibiótico apresentaram perda pouco significativa no módulo de elasticidade, comparadas às amostras processadas por ultrassom e peróxido de hidrogênio que mantiveram a propriedade viscoelástica. Contudo na dose de 12,5 kGy foi observado um aumento no módulo elástico e na viscoelasticidade. Com base nas análises, podemos concluir que o método de processamento com álcool, antibiótico e irradiação demonstrou menor dano, tanto na biomecânica quanto na esterilização, sendo que as amostras irradiadas a 15,0 e 25,0 kGy apresentaram características semelhantes ao controle não irradiado. / The increasing interest of development in less invasive surgical techniques, such as reconstructions of ligament tendon, has led to the increase of the research concerning the use of Allografts sterilized by ionizing radiation. Processing by ionizing radiation is a safe method and leaves no residues, being used as final sterilization. The present study was to evaluate the effects of proposed application of ionizing radiation, produced by 60Co source in human tendon pre-samples processed multiorgans donors obtained through collaboration with tissue banks. The pre-processing of samples given by chemical methods and preserved by freezing at -80 °C. The doses applied in radiation processing were 12.5, 15.0 and 25.0 kGy, each with your corresponding non-irradiated control. The samples were evaluated by means of histological and biomechanical testing, with the purpose of analyzing possible structural and morphological changes. The results showed that the ultrasound processing and hydrogen peroxide caused changes in the morphology of the tissues, which caused damage to the structure, making your samples. In the samples processed by alcohol and antibiotics were not observed damage on the network of collagen by the application of radiation. The results of biomechanical tests showed significant differences between the methods used. The samples processed with alcohol and antibiotics showed negligible loss in modulus of elasticity compared with the samples processed by ultrasound and hydrogen peroxide which kept the viscoelastic property, however in 12.5 kGy dose was observed an increase in elastic modulus and viscoelasticity. Based on the analysis we can conclude that the method of processing with alcohol, antibiotics and irradiation showed less damage, both in biomechanics and sterilization, in the samples irradiated with 15.0 and 25.0 kGy, showing results similar to the non-irradiated control.

Bancos de tumores foram criados para organizar a coleta, arm azenamento e distribuição de amostras biológicas de pacientes oncológicos, favorecendo seu uso nas pesquis as sobre o cân cer. Amostras ade quadas devem ter RNA, DNA e proteínas de boa qualidade. RNA de boa qualidade deve estar íntegro e puro e DNA deve ter boa c oncentração e pur eza. Basea do em norm as in ternacionais, f oi elaborado e implantado um abrangente sistema de controle de qualidade no banco de tumores do Hospital de Câncer de Barretos, que para fins de estudo foi dividido em banco pré-controle de qu alidade (den ominado b anco pré) e em ban co pós- controle de qualidade (denominado banco pós). Objetivando comparar a qualidade das amostras n os dois bancos, atra vés d a extração d e R NA total e d e DNA (utilizando-se homogeneizador de tecidos e Kits), selecionou-se de forma aleatória 200 a mostras tumorais, distribuídas ig ualitariamente entre mama, co lorreto, estômago, pulmão e tireóide, sendo 100 do banco pré e 100 do banco pós. Para se avaliar a influência do tempo de isquemia fria (tempo entre a excisão do e spécime cirúrgico e o congelamento rápido da amostra armazenada) na qualidade do RNA total de amostras tumorais do banco pós, foram coletadas 200 amostras tumorais, distribuídas igualitariamente entre mama, co lorreto, estômago, pulmão e ti reóide, de 100 doadores diferentes, metade com o tempo de isquemia fria (TIF) de até 30 minutos e a o utra metade do mesmo espécime com TIF de 45 minutos. Extraiu-se RNA total dessas amostras (com maceração manual e Trizol) e comparou-se a sua qualidade, através do núm ero de i ntegridade do RNA (RIN), dentr o dos d ois intervalos de tempo e nas diferentes top ografias. Ao c omparar-se amostras com RIN acima de 7 (consideradas ideais para experimentos de microarray), do banco pré e do b anco pó s, for am enc ontrados 73 (73%) no p rimeiro e 87 (87%) no segundo (p=0,013). Ao comparar-se o intervalo de TIF de até 30 minutos com o de 45 minutos, encontrou-se respectivamente 63 (64,3%) e 3 6 (36%) amostras com RNA total intacto, 11 (11,2%) e 17 (17 %) com RNA tot al parcialmente degradado e 24 (24, 5%) e 47 (47%) com RNA t otal degradado (p<0,001). Amostras tireoidianas e colorretais f oram mais sensíveis ao a umento d o T IF (p=0,006 e p=0,03, respectivamente), e as de estômago e pulmão menos sensíveis (p=0,919 e p=0,384, resp ectivamente). Ao comparar-se a s 200 amostras dos dois ban cos, constatou-se que a grande maioria apresentava boa qualidade, porém o banco pós se destacou ao avaliar-se o número de amostras ideais para estudos de microarray, por provável interferência d o TIF, ainda não controlado no banco pr é. Constatou-se também que algumas amostras do banco pré, armazenadas há mais de ci nco anos em freezer a -80ºC, apresentaram excelente qualidade. O presente estudo também mostrou que o TIF é muito importante para a preservação da qualidade do RNA total, por isso, deve-se sempre respeitar o tempo máximo de 30 minutos. Ainda se observou que a de gradação do RNA é tecido dependente e qu e amostras processadas com homogeneizador de tecidos e extraídas com RNeas y Mini Kit apresentaram melhor q ualidade do RNA, qu e as macer adas manualmente e extraídas com Trizol / Tumor banks were created to or ganize the collection, storage and d istribution of biological samples of cancer pa tients, favoring it\'s use in cancer rese arches. Appropriate samples should have good quality of RNA, DNA and p roteins. RNA of good quality should be intact and pure and DNA should have good concentration and pu rity. Ba sed on international sta ndards, we elabo rated and imp lanted an comprehensive s ystem of qu ality control in the tu mor bank of Ba rretos Cancer Hospital, w hich was divided for st udy purposes i n pre bank quality control (denominated pre bank) and post bank qu ality control (denominated post bank). Aiming to compare the quality of the samples in two banks, through the extraction of total RNA and DNA (b y tissue homogenizer and Kits), we se lected 200 tumor samples in a random way, distributed equally among breast, colorectal, stomach, lung and thyroid, being 100 of the pre-bank and 100 of the post bank. To evaluate the influence o f cold ischem ia time (time b etween t he ex cision o f the su rgical specimen and the fast freezing of the stored sample) in the quality of total of RNA tumor sa mples of th e po st bank , we collected 2 00 t umor s amples, distrib uted equally among breast, colorectal, stom ach, lung and th yroid, fro m 100 different donors, half with the cold ischemia time (CIT) up to 30 minutes and the other ha lf of the sam e specimen with CIT exact ly 45 minutes. We ex tracted total RNA of these samples (with manual maceration and T rizol) and c ompared their qu ality, through the RNA integri ty number (RIN), ins ide tw o intervals of time a nd in different topographies. Comparing samples with RIN above 7 (considered ideals for microarray experiments), of the pre bank and of the post bank, we found 73 (73%) in the first and 87 (87%) in the second (p=0,013). Comparing the interval of CIT up to 30 m inutes with the ex actly 45 minutes, we found respectively 63 (64,3%) and 36 (36%) samples with total RNA intact, 11 (11,2%) and 17 (17%) with total RNA partially degraded and 24 (2 4,5%) and 47 (47%) wit h total RNA de graded (p<0,001). Thyroid and colorectal samples were more sensitive to the increase of CIT (p =0,006 and p=0,03, respectively), a nd s tomach and lun g samples less sensitive (p=0,919 and p=0,384, respectively). C omparing the 200 samples from the two b anks, we v erified that the great ma jority had good qu ality; however the post bank stood out the evaluating number of the id eal samples for m icroarray studies, for probable interference of CIT, still n o controlled in the pre bank. We also verified that some samples of the pre bank, stored more than 5 years in freezer at -80 ºC presented e xcellent qu ality. T he stu dy still sho wed that CIT is ver y important to preserve the quality of total RNA, for that, we sh ould always respect the maximum time of 30 minutes. We still observed that the degradation of RNA is tissue dependent and that samples processed with tissue homogenizer and extracted using RNeasy Mini Kit showed better quality of RNA that macerated manually and extracted with Trizol

Bancos de Tecidos de diversas regiões do mundo têm estocado cartilagens humanas obtidas de doadores post mortem para uso em diversos tipos de cirurgias reconstrutivas. Para garantir que tais tecidos não estejam contaminados, estes têm sido esterilizados com radiação ionizante. Entretanto, altas doses de radiação gama podem causar efeitos indesejáveis nos tecidos. No presente trabalho, avaliamos a viabilidade de utilizar duas técnicas, Tomografia por Coerência Óptica (OCT) e Termogravimetria (TGA), para identificar possíveis modificações estruturais causadas na cartilagem costal humana em decorrência dos métodos de preservação e doses de radiação ionizante utilizadas. As cartilagens obtidas de doadores cadavéricos foram congeladas a -70 ºC ou preservadas em glicerol. A seguir, as amostras foram irradiadas por fontes de 60Co com doses de 15, 25 e 50 kGy. Nos resultados de TGA verificamos que as cartilagens preservadas em glicerol e irradiadas com diferentes doses de radiação não apresentaram diferenças estatisticamente significantes quando comparadas ao grupo controle, no que tange a taxa de desidratação do tecido, sendo que o mesmo não ocorre com cartilagens congeladas a -70 ºC e irradiadas com doses de 15 kGy. Em relação ao uso da técnica de OCT, por meio do cálculo do coeficiente de atenuação óptica total, verificamos que doses de 15 kGy promovem a criação de ligações cruzadas entre as fibrilas de colágeno, corroborando os resultados de TGA. Ainda, os valores do coeficiente de atenuação óptica total são diretamente proporcionais à tensão de ruptura das cartilagens, o que nos possibilitará, em um futuro próximo, predizer a qualidade de um enxerto sem a necessidade de perda de material biológico, visto ser o OCT um método não destrutivo. Por meio das imagens de PS-OCT podemos verificar que as doses de radiação utilizadas para esterilizar as amostras não provocam danos à rede de colágeno a ponto de que sua birrefringência seja perdida. Assim, o TGA e OCT são técnicas que podem ser utilizadas por bancos de tecidos de forma a verificar a qualidade dos tecidos antes de serem transplantados em pacientes. / Tissue Banks around the world have stored human cartilages obtained from post mortem donors for use in several kinds of reconstructive surgeries. To ensure that such tissues are not contaminated, they have been sterilized with ionizing radiation. However, high doses of gamma radiation may cause undesirable changes in the tissues. In this work, we evaluated the possibility of use Optical Coherence Tomography (OCT) and Thermogravimetric Analysis (TGA) to identify possible structural modifications caused by both preservation methods of cartilage and gamma irradiation doses. Cartilages were obtained from cadaveric donors and were frozen at -70 ºC or preserved in glycerol. Irradiation was performed by 60Co source with doses of 15, 25 and 50 kGy. Our TGA results showed that glycerolized cartilages irradiated with different doses of radiation does not presented statistical differences when compared to the control group for the dehydration rate. However, the same was not observed for deep-fronzen cartilages irradiated with 15 kGy. The results of OCT associated to total optical attenuation coefficient showed that doses of 15 kGy promote cross-link between collagen fibrils, corroborating the results obtained from TGA. Moreover, total optical attenuation coefficient values are proportionals to stress at break of cartilages, what will be very useful in a near future to predict the quality of the allografts, without unnecessary loss of biological tissue, once OCT is a nondestructive technique. By PS-OCT images, we found that high doses of ionizing radiation does not promote sufficient impairments to promote complete loss of tissue birefringence. Thus, TGA and OCT are techniques that can be used for tissue banks to verify tissue quality before its transplant.

Bancos de tumores foram criados para organizar a coleta, arm azenamento e distribuição de amostras biológicas de pacientes oncológicos, favorecendo seu uso nas pesquis as sobre o cân cer. Amostras ade quadas devem ter RNA, DNA e proteínas de boa qualidade. RNA de boa qualidade deve estar íntegro e puro e DNA deve ter boa c oncentração e pur eza. Basea do em norm as in ternacionais, f oi elaborado e implantado um abrangente sistema de controle de qualidade no banco de tumores do Hospital de Câncer de Barretos, que para fins de estudo foi dividido em banco pré-controle de qu alidade (den ominado b anco pré) e em ban co pós- controle de qualidade (denominado banco pós). Objetivando comparar a qualidade das amostras n os dois bancos, atra vés d a extração d e R NA total e d e DNA (utilizando-se homogeneizador de tecidos e Kits), selecionou-se de forma aleatória 200 a mostras tumorais, distribuídas ig ualitariamente entre mama, co lorreto, estômago, pulmão e tireóide, sendo 100 do banco pré e 100 do banco pós. Para se avaliar a influência do tempo de isquemia fria (tempo entre a excisão do e spécime cirúrgico e o congelamento rápido da amostra armazenada) na qualidade do RNA total de amostras tumorais do banco pós, foram coletadas 200 amostras tumorais, distribuídas igualitariamente entre mama, co lorreto, estômago, pulmão e ti reóide, de 100 doadores diferentes, metade com o tempo de isquemia fria (TIF) de até 30 minutos e a o utra metade do mesmo espécime com TIF de 45 minutos. Extraiu-se RNA total dessas amostras (com maceração manual e Trizol) e comparou-se a sua qualidade, através do núm ero de i ntegridade do RNA (RIN), dentr o dos d ois intervalos de tempo e nas diferentes top ografias. Ao c omparar-se amostras com RIN acima de 7 (consideradas ideais para experimentos de microarray), do banco pré e do b anco pó s, for am enc ontrados 73 (73%) no p rimeiro e 87 (87%) no segundo (p=0,013). Ao comparar-se o intervalo de TIF de até 30 minutos com o de 45 minutos, encontrou-se respectivamente 63 (64,3%) e 3 6 (36%) amostras com RNA total intacto, 11 (11,2%) e 17 (17 %) com RNA tot al parcialmente degradado e 24 (24, 5%) e 47 (47%) com RNA t otal degradado (p<0,001). Amostras tireoidianas e colorretais f oram mais sensíveis ao a umento d o T IF (p=0,006 e p=0,03, respectivamente), e as de estômago e pulmão menos sensíveis (p=0,919 e p=0,384, resp ectivamente). Ao comparar-se a s 200 amostras dos dois ban cos, constatou-se que a grande maioria apresentava boa qualidade, porém o banco pós se destacou ao avaliar-se o número de amostras ideais para estudos de microarray, por provável interferência d o TIF, ainda não controlado no banco pr é. Constatou-se também que algumas amostras do banco pré, armazenadas há mais de ci nco anos em freezer a -80ºC, apresentaram excelente qualidade. O presente estudo também mostrou que o TIF é muito importante para a preservação da qualidade do RNA total, por isso, deve-se sempre respeitar o tempo máximo de 30 minutos. Ainda se observou que a de gradação do RNA é tecido dependente e qu e amostras processadas com homogeneizador de tecidos e extraídas com RNeas y Mini Kit apresentaram melhor q ualidade do RNA, qu e as macer adas manualmente e extraídas com Trizol / Tumor banks were created to or ganize the collection, storage and d istribution of biological samples of cancer pa tients, favoring it\'s use in cancer rese arches. Appropriate samples should have good quality of RNA, DNA and p roteins. RNA of good quality should be intact and pure and DNA should have good concentration and pu rity. Ba sed on international sta ndards, we elabo rated and imp lanted an comprehensive s ystem of qu ality control in the tu mor bank of Ba rretos Cancer Hospital, w hich was divided for st udy purposes i n pre bank quality control (denominated pre bank) and post bank qu ality control (denominated post bank). Aiming to compare the quality of the samples in two banks, through the extraction of total RNA and DNA (b y tissue homogenizer and Kits), we se lected 200 tumor samples in a random way, distributed equally among breast, colorectal, stomach, lung and thyroid, being 100 of the pre-bank and 100 of the post bank. To evaluate the influence o f cold ischem ia time (time b etween t he ex cision o f the su rgical specimen and the fast freezing of the stored sample) in the quality of total of RNA tumor sa mples of th e po st bank , we collected 2 00 t umor s amples, distrib uted equally among breast, colorectal, stom ach, lung and th yroid, fro m 100 different donors, half with the cold ischemia time (CIT) up to 30 minutes and the other ha lf of the sam e specimen with CIT exact ly 45 minutes. We ex tracted total RNA of these samples (with manual maceration and T rizol) and c ompared their qu ality, through the RNA integri ty number (RIN), ins ide tw o intervals of time a nd in different topographies. Comparing samples with RIN above 7 (considered ideals for microarray experiments), of the pre bank and of the post bank, we found 73 (73%) in the first and 87 (87%) in the second (p=0,013). Comparing the interval of CIT up to 30 m inutes with the ex actly 45 minutes, we found respectively 63 (64,3%) and 36 (36%) samples with total RNA intact, 11 (11,2%) and 17 (17%) with total RNA partially degraded and 24 (2 4,5%) and 47 (47%) wit h total RNA de graded (p<0,001). Thyroid and colorectal samples were more sensitive to the increase of CIT (p =0,006 and p=0,03, respectively), a nd s tomach and lun g samples less sensitive (p=0,919 and p=0,384, respectively). C omparing the 200 samples from the two b anks, we v erified that the great ma jority had good qu ality; however the post bank stood out the evaluating number of the id eal samples for m icroarray studies, for probable interference of CIT, still n o controlled in the pre bank. We also verified that some samples of the pre bank, stored more than 5 years in freezer at -80 ºC presented e xcellent qu ality. T he stu dy still sho wed that CIT is ver y important to preserve the quality of total RNA, for that, we sh ould always respect the maximum time of 30 minutes. We still observed that the degradation of RNA is tissue dependent and that samples processed with tissue homogenizer and extracted using RNeasy Mini Kit showed better quality of RNA that macerated manually and extracted with Trizol

Bancos de Tecidos de diversas regiões do mundo têm estocado cartilagens humanas obtidas de doadores post mortem para uso em diversos tipos de cirurgias reconstrutivas. Para garantir que tais tecidos não estejam contaminados, estes têm sido esterilizados com radiação ionizante. Entretanto, altas doses de radiação gama podem causar efeitos indesejáveis nos tecidos. No presente trabalho, avaliamos a viabilidade de utilizar duas técnicas, Tomografia por Coerência Óptica (OCT) e Termogravimetria (TGA), para identificar possíveis modificações estruturais causadas na cartilagem costal humana em decorrência dos métodos de preservação e doses de radiação ionizante utilizadas. As cartilagens obtidas de doadores cadavéricos foram congeladas a -70 ºC ou preservadas em glicerol. A seguir, as amostras foram irradiadas por fontes de 60Co com doses de 15, 25 e 50 kGy. Nos resultados de TGA verificamos que as cartilagens preservadas em glicerol e irradiadas com diferentes doses de radiação não apresentaram diferenças estatisticamente significantes quando comparadas ao grupo controle, no que tange a taxa de desidratação do tecido, sendo que o mesmo não ocorre com cartilagens congeladas a -70 ºC e irradiadas com doses de 15 kGy. Em relação ao uso da técnica de OCT, por meio do cálculo do coeficiente de atenuação óptica total, verificamos que doses de 15 kGy promovem a criação de ligações cruzadas entre as fibrilas de colágeno, corroborando os resultados de TGA. Ainda, os valores do coeficiente de atenuação óptica total são diretamente proporcionais à tensão de ruptura das cartilagens, o que nos possibilitará, em um futuro próximo, predizer a qualidade de um enxerto sem a necessidade de perda de material biológico, visto ser o OCT um método não destrutivo. Por meio das imagens de PS-OCT podemos verificar que as doses de radiação utilizadas para esterilizar as amostras não provocam danos à rede de colágeno a ponto de que sua birrefringência seja perdida. Assim, o TGA e OCT são técnicas que podem ser utilizadas por bancos de tecidos de forma a verificar a qualidade dos tecidos antes de serem transplantados em pacientes. / Tissue Banks around the world have stored human cartilages obtained from post mortem donors for use in several kinds of reconstructive surgeries. To ensure that such tissues are not contaminated, they have been sterilized with ionizing radiation. However, high doses of gamma radiation may cause undesirable changes in the tissues. In this work, we evaluated the possibility of use Optical Coherence Tomography (OCT) and Thermogravimetric Analysis (TGA) to identify possible structural modifications caused by both preservation methods of cartilage and gamma irradiation doses. Cartilages were obtained from cadaveric donors and were frozen at -70 ºC or preserved in glycerol. Irradiation was performed by 60Co source with doses of 15, 25 and 50 kGy. Our TGA results showed that glycerolized cartilages irradiated with different doses of radiation does not presented statistical differences when compared to the control group for the dehydration rate. However, the same was not observed for deep-fronzen cartilages irradiated with 15 kGy. The results of OCT associated to total optical attenuation coefficient showed that doses of 15 kGy promote cross-link between collagen fibrils, corroborating the results obtained from TGA. Moreover, total optical attenuation coefficient values are proportionals to stress at break of cartilages, what will be very useful in a near future to predict the quality of the allografts, without unnecessary loss of biological tissue, once OCT is a nondestructive technique. By PS-OCT images, we found that high doses of ionizing radiation does not promote sufficient impairments to promote complete loss of tissue birefringence. Thus, TGA and OCT are techniques that can be used for tissue banks to verify tissue quality before its transplant.