Biological parameters for quality evaluation of allografts from the Brazilian National Institute of Traumatology and Orthopedics tissue bank.

Bone allografts are clinically used in a variety of surgical procedures, and tissue banks are responsible for harvesting, processing, quality testing, storing, and delivering these materials for transplantation. In tissue banks, the bone is processed for the removal of all organic content, remaining only the tissue structure (scaffold). However, several studies have shown that even after using different processing methods, viable cells, functional proteins, and DNA may still persist in the tissue, which constitute the main causes of graft rejection. Therefore, the objective of this study was to establish techniques and biological parameters for quality validation of allografts. To this end, we propose the use of 3 combined methods such as microscopy, histology, and molecular biology techniques to evaluate the quality of allografts harvested and processed by the Brazilian National Institute of Traumatology and Orthopedics (INTO) tissue bank according to the donation criteria of the Brazilian National Health Surveillance Agency and the Brazilian National Transplant System. Bone fragments from different processing stages showed no viable cells on histology, an intact extracellular matrix on scanning electron microscopy, and gradual reduction in DNA amount. Different techniques were used to demonstrate the quality of allografts produced by the INTO tissue bank and to establish biological parameters for ensuring the safety and quality of these products. Future studies need to be undertaken to assess and validate the efficacy of the decellularization process in larger bone grafts with diverse architectural configurations.

Family refusal of skin donation for transplantation: trends and associated factors / Rechazo familiar de donación de piel para trasplante: tendencias y factores asociados / Recusa familiar de doação de pele para transplante: tendência e fatores associados

ABSTRACT Objectives: to analyze the trends and factors associated with family refusal of skin donation for transplantation. Methods: this cross-sectional study was conducted in the State of São Paulo, with family authorization terms collected from 2001 to 2020. The variables analyzed included year, age, gender, cause of death, and type of institution. Data were analyzed using linear and multiple logistic regression, with the Odds Ratio estimated at p<0.05 for statistical significance. Results: 1,355 individuals refused skin donation. The trend of refusals decreased between 2001 and 2009 in the age groups of 0-11 years and 12-19 years, but increased in the group aged ≥60 years. This trend continued to decrease in the 0-11 years group from 2010 to 2020, and increased in the 20-40 years group. Males and the age groups of 20-40 years, 41-59 years, and ≥60 years exhibited 27%, 34%, 47%, and 53% lower chances of refusal, respectively. Conclusions: there is an urgent need for measures to mitigate the high number of refusals associated with skin donation.

RESUMEN Objetivos: analizar la tendencia y los factores asociados al rechazo familiar de la donación de piel para trasplante. Métodos: estudio transversal desarrollado en el Estado de São Paulo con los términos de autorización familiar firmados entre 2001 y 2020. Las variables analizadas fueron: año, edad, sexo, causa del deceso y tipo de institución. Los datos fueron analizados mediante regresión lineal y logística múltiple, con la estimación del Odds Ratio, adoptando p<0,05 como significancia estadística. Resultados: 1.355 individuos rechazaron la donación de piel. La tendencia de rechazos fue decreciente (2001-2009) en los grupos de edad de 0-11 años y de 12-19 años, y creciente en el grupo de ≥60 años. La tendencia siguió siendo decreciente (2010-2020) en el grupo de edad de 0-11 años y creciente en el de 20-40 años. El género masculino y los grupos de edad de 20-40 años, 41-59 años y ≥60 años presentaron, respectivamente, 27%, 34%, 47% y 53% menos probabilidades de rechazo. Conclusiones: es urgente la necesidad de medidas que busquen mitigar el alto número de rechazos asociados a la donación de piel.

RESUMO Objetivos: analisar a tendência e os fatores associados à recusa familiar de doação de pele para transplante. Métodos: estudo transversal desenvolvido no Estado de São Paulo com os termos de autorização familiar firmados entre 2001 e 2020. As variáveis analisadas foram: ano, idade, sexo, causa do óbito e tipo da instituição. Os dados foram analisados por meio de regressão linear e logística múltipla, com o Odds Ratio estimado, adotando-se p<0,05 como significância estatística. Resultados: 1.355 indivíduos recusaram a doação de pele. A tendência de recusas foi decrescente (2001-2009) nas faixas etárias de 0-11 anos e de 12-19 anos e crescente na faixa de ≥60 anos. A tendência permaneceu decrescente (2010-2020) na faixa etária de 0-11 anos e crescente na de 20-40 anos. O sexo masculino e as faixas etárias de 20-40 anos, 41-59 anos e ≥60 anos apresentaram, respectivamente, 27%, 34%, 47% e 53% menores chances de recusa. Conclusões: é urgente a necessidade de medidas que visem atenuar o alto número de recusas associadas à doação de pele.

A novel method to pack cryopreserved amniotic membrane for wound dressing-the pathway through validation of a new biological product.

Amniotic membrane (AM) has been widely used as a biological dressing for many pathologies and illnesses worldwide, and products derived from this tissue have been commercially available in several countries. In Brazil, regulatory agencies have recently authorized its clinical use as a non-experimental therapy for burns, diabetic and venous stasis ulcers, and intrauterine adhesions. In this study, we present our pathway through validating the first available service in the country of AM cryopreservation, with a protocol for long-term storage in high-efficiency nitrogen cryogenic freezers and a specific way of packing the tissue for optimal clinical handling and efficient storage space utilization while preserving live cells and the tissue's biological properties. Using gauze as support, cryoprotectant dimethyl sulfoxide and product presentation as a multilayer roll exhibited the best cell viability results and maintained the tissue integrity and presence of stem/progenitor cells. Essential proteins involved in tissue regeneration and immune and antimicrobial control were detected from the secretome of cryopreserved tissue similar to fresh tissue. Furthermore, immunogenic markers, such as human leukocyte antigens, were detected at very low levels in the tissue, confirming their low immunogenicity. Finally, we demonstrate that the tissue can be kept under refrigerated conditions for up to 7 d for further use, maintaining sterility and considerable cell viability. Our cryopreservation and storage protocol kept the AM viable for at least 20 months. In conclusion, this study enabled us to determine a novel efficient protocol for long-term AM preservation for future clinical applications.

The Significance of the Nucleic Acid Test (NAT) to Prevent Viral Contamination in Musculoskeletal Tissue Transplantation.

Objective The present study aims to highlight the significance of the nucleic acid test (NAT) for musculoskeletal tissue donation and to compare the sensitivity of this test on the different available platforms. Method The present study is a retrospective survey in a human tissue bank database and an integrative literature review encompassing the last 10 years. The PubMed portal and the SCOPUS, CINAHL, and Web of Science databases were queried for articles. Results We found no specific studies on the use and sensitivity of NAT in braindead tissue donors. The information presented in the present study consists of specific contents intended for the Brazilian Blood Transfusion Network (Hemorrede Transfusional Nacional, in Portuguese) and internal retrospective data from a tissue bank located at a city in the state of São Paulo, Brazil. Conclusions The NAT is effective in blood samples from living patients. However, since biochemical reactions in braindead patients can be different, specific research, platforms, or both are crucial to tissue banks.

The Significance of the Nucleic Acid Test (NAT) to Prevent Viral Contamination in Musculoskeletal Tissue Transplantation / A importância da realização do teste de ácidos nucleicos (NAT) para a prevenção de contaminação viral nos transplantes de tecidos musculoesqueléticos

Abstract Objective The present study aims to highlight the significance of the nucleic acid test (NAT) for musculoskeletal tissue donation and to compare the sensitivity of this test on the different available platforms. Method The present study is a retrospective survey in a human tissue bank database and an integrative literature review encompassing the last 10 years. The PubMed portal and the SCOPUS, CINAHL, and Web of Science databases were queried for articles. Results We found no specific studies on the use and sensitivity of NAT in braindead tissue donors. The information presented in the present study consists of specific contents intended for the Brazilian Blood Transfusion Network (Hemorrede Transfusional Nacional, in Portuguese) and internal retrospective data from a tissue bank located at a city in the state of São Paulo, Brazil. Conclusions The NAT is effective in blood samples from living patients. However, since biochemical reactions in braindead patients can be different, specific research, platforms, or both are crucial to tissue banks.

Resumo Objetivo Evidenciar a importância da realização do teste de ácido nucleico (NAT, na sigla em inglês) para doação de tecidos musculoesqueléticos, assim como comparar a sensibilidade deste exame nas diferentes plataformas existentes no mercado. Método Trata-se de um levantamento retrospectivo no banco de dados de um determinado Banco de Tecidos Humanos e de uma revisão integrativa da literatura, operacionalizada nos últimos 10 anos. As buscas de artigos ocorreram no portal PubMed e nas bases de dados SCOPUS, CINAHL e Web of Science. Resultados Não foram encontrados estudos específicos sobre a utilização e a sensibilidade do exame NAT em pacientes doadores de tecidos com morte encefálica (ME), sendo as informações apresentadas no presente estudo conteúdos específicos destinados à Hemorrede Transfusional Nacional e aos dados retrospectivos internos de um Banco de Tecidos do interior do estado de São Paulo, Brasil. Conclusões O exame NAT se apresenta efetivo em amostras de sangue de pacientes vivos. Porém, reações bioquímicas em pacientes com condições de ME podem se apresentar de formas diferenciadas, tornando-se indispensáveis a realização de pesquisas específicas e/ou a indicação de plataformas aos Bancos de Tecidos.

Resultados post operatorios en pacientes con prótesis total de revisión de cadera mas injerto óseo cadavérico de banco de tejidos en el Hospital de Clínicas en el periodo 2017 al 2020 / Postoperative results in patients with total hip revision arthroplasty and cadaveric bone graft of tissue bank at the Hospital de Clínicas in the period 2017 to 2020

Introducción: La artroplastia total de cadera de revisión junto con la utilización de injerto óseo cadavérico de banco de tejidos es una alternativa de tratamiento eficaz ante los procedimientos de reemplazo con déficit óseo femoral o acetabular. El presente estudio, analizó los resultados post operatorios en pacientes intervenidos quirúrgicamente en el Hospital de Clínicas. Materiales y métodos: Estudio descriptivo. Muestreo no probabilístico de casos consecutivos atendidos en la Cátedra de Ortopedia y Traumatología de la Facultad de Ciencias Médicas (U.N.A). Fueron captados pacientes con indicación de artroplastia total de cadera de revisión (ATCR) más injerto óseo cadavérico de banco de tejidos, entre diciembre 2017 - octubre 2020, previo consentimiento informado. Se relevaron datos cuya caracterización fue posible mediante un instrumento establecido previamente, ingresado en base Excel. Resultados: 12 pacientes con déficit óseo femoral y acetabular han sido tratados con artroplastia total de cadera de revisión más injerto óseo cadavérico de banco de tejidos, en donde además de la funcionalidad y a través de seguimientos radiológicos se ha determinado la osteointegración total de los aloinjertos. La relación masculino-femenina fue 1/1, siendo el diagnóstico preoperatorio para la colocación de la prótesis de revisión más injerto óseo cadavérico de banco de tejidos el aflojamiento séptico en un 75%. El motivo de consulta más frecuente fue dolor y la secreción y el tiempo quirúrgico fue en promedio de 116 minutos. Conclusión: La artroplastia total de cadera de revisión más aloinjerto presenta óptimos resultados en relación a la osteointegración total y funcionalidad.

Introduction: Revision total hip arthroplasty with use of cadaveric bone graft is an effective treatment for replacement procedures in patients with bone loss femoral or acetabular. The present study analyzed the postoperative results in patients who underwent surgery at the Hospital de Clínicas. Materials and methods: Descriptive study. Non-probabilistic sampling of consecutive cases treated at the Department of Orthopedics and Traumatology of Hospital de Clínicas. Data of patients with indication for revision total hip arthroplasty plus cadaveric bone graft from a tissue bank with prior informed consent,were analyzed between December 2017 and October 2020. Results: 12 patients with femoral and acetabular bone deficits have been treated with revision total hip arthroplasty plus cadaveric bone graft from a tissue bank. A radiological and functional follow up was made. The male-female ratio was 1/1, with the preoperative diagnosis for placement of the revision prosthesis plus cadaveric bone graft,from the tissue bank,was septic loosening in 75%. The most frequent symptom was pain, all revision total hip arthroplasties were performed through a posterior approach, and the surgical time was an average of 116 minutes,using a later approach in all cases.Two intraoperative fracture was presented, and were resolved. Conclusion: Revision total hip arthroplasty plus allograft presents optimal results in relation to total osseointegration and functionality.

Evaluation of Histological Properties of Human Meniscal Grafts Stored in a Tissue Bank.

Objectives The present paper aims to evaluate and compare the histological features of fresh and frozen menisci stored in a tissue bank for 1 month and for 5 years. Methods The meniscal grafts were subjected to a histological study. A total of 10 menisci were evaluated; 2 were frozen for 5 years, 4 were frozen for 1 month, and 4 were fresh, recently harvested specimens. Histological properties were evaluated in sections stained with hematoxylin and eosin and Masson trichrome methods. Results The menisci frozen for 1 month showed partially preserved collagen fiber structure and no significant hydropic tissue degeneration. The menisci frozen for 5 years presented an evident dissociation of collagen fibers and multiple foci of hydropic degeneration. Discussion Degeneration was much more significant in menisci stored for 5 years, indicating that a long freezing period results in substantial progression of tissue deterioration. This may suggest that the 5-year period, considered the maximum time for graft storage before transplant, is too long. Conclusion Grafts stored for 1 month showed a slight degenerative change in collagen fibers, whereas menisci frozen for 5 years presented significant tissue degeneration.

Evaluation of Histological Properties of Human Meniscal Grafts Stored in a Tissue Bank / Avaliação das propriedades histológicas de enxertos meniscais humanos armazenados em banco de tecido

Abstract Objectives The present paper aims to evaluate and compare the histological features of fresh and frozen menisci stored in a tissue bank for 1 month and for 5 years. Methods The meniscal grafts were subjected to a histological study. A total of 10 menisci were evaluated; 2 were frozen for 5 years, 4 were frozen for 1 month, and 4 were fresh, recently harvested specimens. Histological properties were evaluated in sections stained with hematoxylin and eosin and Masson trichrome methods. Results The menisci frozen for 1 month showed partially preserved collagen fiber structure and no significant hydropic tissue degeneration. The menisci frozen for 5 years presented an evident dissociation of collagen fibers and multiple foci of hydropic degeneration. Discussion Degeneration was much more significant in menisci stored for 5 years, indicating that a long freezing period results in substantial progression of tissue deterioration. This may suggest that the 5-year period, considered the maximum time for graft storage before transplant, is too long. Conclusion Grafts stored for 1 month showed a slight degenerative change in collagen fibers, whereas menisci frozen for 5 years presented significant tissue degeneration.

Resumo Objetivos Avaliar e comparar as características histológicas de meniscos frescos e meniscos congelados armazenados em banco de tecidos por 1 mês e por 5 anos. Métodos Foi feito um estudo histológico com enxertos meniscais. Avaliamos 10 meniscos, sendo 2 que ficaram armazenados sob congelamento por 5 anos, 4 armazenados congelados por 1 mês, e 4 frescos, recém captados. Foram feitos cortes histológicos corados com hematoxilina e eosina e Tricrômico de Masson, para avaliação das propriedades histológicas. Resultados Os meniscos congelados por 1 mês apresentaram preservação parcial da estrutura das fibras colágenas, sem degeneração hidrópica significativa do tecido. Nos meniscos congelados por 5 anos, observamos dissociação evidente das fibras colágenas, com presença de múltiplos focos de degeneração hidrópica. Discussão Encontramos degeneração bem mais significativa nos meniscos armazenados por 5 anos, o que indica que o longo período de congelamento leva à progressão significativa da degeneração do tecido. Isto pode sugerir que o período de 5 anos, considerado período máximo que o enxerto pode permanecer armazenado antes de ser transplantado, é um período muito longo. Conclusão Nos enxertos armazenados por 1 mês, existiu apenas discreta alteração degenerativa das fibras colágenas, enquanto que nos meniscos com 5 anos de congelamento foi observada degeneração significativa do tecido. Tibiais

Implementation and certification of ISO 9001:2015 seal in human tissue bank HCFMRP-USP.

The Quality Management System has been a management technology applied to guarantee the quality of processes and products of a given organization. Thus, ISO 9001:2015 certification assists organizations that want to develop, implement, maintain, and improve a quality management system to enable process improvements and assessments to meet the needs of its customers. This experience report addresses the implementation and certification of the ISO 9001:2015 quality management seal at the Human Tissue Bank of the Ribeirão Preto Clinical Hospital (HCRP) at the University of São Paulo (USP) at Ribeirão Preto Medical School (FMRP)-Brazil. In 2018, the Human Tissue Bank, in partnership with HCRP, received consultancies from PRIMEMODE. The consulting consisted of visits that elucidated the processes to the employees of the Tissue Bank, enabling them to clarify the applicability of all items of the standard. In March 2019, the Tissue Bank received the audit visit and was certified with the ISO 9001:2015 Seal by the Carlos Alberto Vanzolini Foundation. The Tissue Bank aims to provide human tissues for transplantation and research from organ donors. It has a complex physical structure within the required health and legal standards. Also, now it has a quality management seal. The certification guarantees the process organization and the high quality standard of the supplied tissues.

A Biobank of Stem Cells of Human Exfoliated Deciduous Teeth: Overview of Applications and Developments in Brazil.

A biobank is an organized collection of biological human material and its associated information stored for research according to regulations under institutional responsibility, without commercial purposes, being a mandatory and strategical activity for research, regenerative medicine, and innovation. Stem cells have largely been employed in research and frequently stored in biobanks, which have been used as an essential source of biological materials. Stem cells of human exfoliated deciduous teeth (SHED) are stem cells which have a high multipotency and can be easily obtained. Besides, this extremely accessible tissue has advantages with respect to storage, as the SHED obtained in childhood can be used in later life, which implies the necessity for the creation and regulation of biobanks. The proper planning for the creation of a biobank includes knowledge of the material types to be stored, requirements regarding handling and storage conditions, storage time, and room for the number of samples. Thus, this study aimed to establish an overview of the development of a SHED biobank. Ethical and legal standardization, current applications, specific orientations, and challenges for the implementation of a SHED biobank were discussed. Through this overview, we hope to encourage further studies to use SHED biobanks.

Detection and quantification of human immunodeficiency virus and hepatitis C virus in cadaveric tissue donors using different molecular tests.

BACKGROUND: Tissues from cadaveric donors are used in several clinical circumstances, and the transmission of infectious diseases has been reported. Cadaveric donor (CD) blood sample analysis is challenging due to its poor quality. However, studies have demonstrated the usefulness of molecular based methods, and the lack of studies using available commercial molecular tests was reported. OBJECTIVE: The aim of this study was to evaluate the performance, specificity, sensitivity, and accuracy of different commercial molecular tests for HIV and HCV detection and quantification in CD through spiked samples. STUDY DESIGN: 20 CD and 20 blood donor samples were tested using 1,000 copies/mL and 1,000 IU/mL of lyophilized standards of HIV and HCV, respectively. Samples were analyzed by different molecular kits: XPERT HCV Viral Load and HIV-1 (Cepheid), COBAS® TaqMan® HIV-1 and COBAS® TaqMan® HCV Test, v2.0 (Roche), and artus® HI Virus-1 QS-RGQ and artus® HCV RG RT-PCR Kit (Qiagen). RESULTS: HIV and HCV in CD were detected by RT-PCR-based quantitative kits. The tests performed by the Cepheid and the Roche kits showed the most accurate, sensitive and specific results, however, a wide variability between the assays and kits was observed. The Qiagen kits did not demonstrate satisfactory results. CONCLUSIONS: CD evaluation showed great variability. The Cepheid and Roche kits were more sensitive for detecting HIV on CD and Cepheid was the most efficient kit for HCV quantification in CD. The Roche and Cepheid kits can be used to screen tissue donors for HIV and HCV.

Anaerobic bioburden in transport solution of human cardiovascular tissues.

Although reports of infections caused by anaerobes after tissue transplantation are uncommon, contamination of allografts may result in substantial complications. Anaerobic incubation and testing of organ transport solution (TS) are not routine. The aim of this study was to determine the bioburden of strict anaerobic bacteria and oxygen tension of heart-TS. Forty TS from different donors were evaluated cultured using membrane filtration (MF), direct inoculation on broth and automated blood culture bottle (ABCB). Bacterial identification was performed by MALDI-TOF. The transport conditions were simulated to verify the bacterial recovery. A sterile bag fulfilled with 250 ml-1 of sterile saline was spiked with 100 CFU ml-1 of Clostridium perfringens and the fluid recovered 0 h, 1 h, 2 h, 6 h, 12 h, 24 h and 48 h for culture and oxygen measurement. Strict anaerobic bacteria were not isolated in heart-TS. The recovery of C.perfringens spiked in heart-TS was 100% using automated blood culture bottles. MF method detected >100 CFU only after 6 h of spiking. The manual culture was not able to recover C.perfringens after the process. The percentage of O2 measures varied from 77.6 to 87.9%. MF or ABCB are better than direct inoculation for recovery of anaerobes from heart-TS. Although all samples from heart donors were negative for anaerobes (probably due to low incidence of contamination), C.perfringens were all recovered in the simulated transport condition.

Comparative Study of the Osteointegration of Irradiated and Non-irradiated Bone Grafts Used in Patients with Revision Hip Arthroplasty.

Objective To evaluate and compare the osteointegration of irradiated and non-irradiated frozen bone grafts used in 21 patients undergoing revision hip arthroplasty procedures with the Exeter technique. Methods A retrospective study of 21 patients undergoing revision hip arthroplasty with the Exeter technique using bone tissues treated or not with gamma radiation between 2013 and 2014. The patients were divided into two groups according to the use of grafts treated or not with ionizing radiation (gamma rays); as such, these groups were classified as irradiated or non-irradiated. The osteointegration results determined by radiographic analysis of these grafts were compared in the postoperative period of 6 and 12 months. Results Comparing the graft osteointegration in all patients at 6 and 12 months postoperatively, we noticed a significant difference in the radiographic evaluations in this period ( p = 0.031). Out of the patients studied, 7 were from the irradiated group, and 14 belonged to the non-irradiated group. No statistically significant differences were observed ( p = 0.804) regarding osteointegration when we compared the irradiated and non-irradiated groups. Conclusion There was no significant difference in the use of irradiated or non-irradiated grafts in revision hip arthroplasty procedures with the Exeter technique.

Comparative Study of the Osteointegration of Irradiated and Non-irradiated Bone Grafts Used in Patients with Revision Hip Arthroplasty / Estudo comparativo da osteointegração de enxertos ósseos irradiados e não irradiados utilizados em pacientes com revisão de artroplastia do quadril

Abstract Objective To evaluate and compare the osteointegration of irradiated and nonirradiated frozen bone grafts used in 21 patients undergoing revision hip arthroplasty procedures with the Exeter technique. Methods A retrospective study of 21 patients undergoing revision hip arthroplasty with the Exeter technique using bone tissues treated or not with gamma radiation between 2013 and 2014. The patients were divided into two groups according to the use of grafts treated or not with ionizing radiation (gamma rays); as such, these groups were classified as irradiated or non-irradiated. The osteointegration results determined by radiographic analysis of these grafts were compared in the postoperative period of 6 and 12months. Results Comparing the graft osteointegration in all patients at 6 and 12months postoperatively, we noticed a significant difference in the radiographic evaluations in this period (p = 0.031). Out of the patients studied, 7 were from the irradiated group, and 14 belonged to the non-irradiated group. No statistically significant differences were observed (p = 0.804) regarding osteointegration when we compared the irradiated and non-irradiated groups. Conclusion There was no significant difference in the use of irradiated or nonirradiated grafts in revision hip arthroplasty procedures with the Exeter technique.

Resumo Objetivo Avaliar e comparar a osteointegração dos enxertos ósseos congelados irradiados e não irradiados utilizados em 21 pacientes submetidos a revisão de prótese do quadril pela técnica Exeter. Métodos Foi realizado estudo retrospectivo de 21 pacientes submetidos a revisão de artroplastia do quadril pela técnica Exeter comutilização de tecidos ósseos tratados ou não com radiação gama no período entre 2013 e 2014. Dividimos os pacientes em dois grupos, de acordo com o uso do enxerto tratado ou não com radiação ionizante (raios gama), que foram, portanto, classificados como: grupo irradiado e não irradiado. Os resultados da osteointegração por análise radiográfica destes enxertos foram comparados no pós-cirúrgico de 6 e 12 meses. Resultados Quando comparamos a osteointegração dos enxertos no pós-cirúrgico de 6 e 12 meses de todos os pacientes, notamos que houve diferença significativa entre as avaliações radiográficas neste período (p= 0,031). Dos pacientes estudados, 7 pertenciam ao grupo irradiado, e 14, ao grupo não irradiado. Não foram observadas diferenças estatisticamente significativas (p= 0,804) quando a osteointegração entre os grupos irradiados e não irradiados foi comparada. Conclusão Não houve diferença significativa no uso de enxerto irradiado e não irradiado nas revisões de artroplastias do quadril pela técnica Exeter.

Conventional culture method and qPCR using 16S rDNA for tissue bank: a comparison using a model of cardiac tissue contamination.

Real-time polymerase chain reaction (qPCR) using 16S rDNA is an alternative to conventional culture-based tests. The aim of this study was to compare the conventional culture method with qPCR using 16S rDNA in a model of cardiac tissue contamination. Samples of cardiac tissue for artificial contamination with Escherichia coli and control samples were submitted for DNA extraction, which was conducted by selective and alkaline lysis and purification steps. A standard curve for 16S rDNA was constructed to determine the efficiency and analytical sensitivity of the assay in concentrations from 106 to 102 c.f.u. ml-1 using TaqMan Master Mix. 16S rDNA was detected in all contaminated samples; however, it was not detected in the the final washing step solution of the sample with a bioburden of 102 c.f.u. ml-1. Using qPCR is a potential alternative to conventional culture for microbiological safety testing of allograft tissues for biobanking, reducing the time and labour input required.

Analysis of potential contamination factors in musculoskeletal tissues.

Tissue Banks have become the main source for bone grafts, due to preference for homologous tissues. Notwithstanding the use of aseptic techniques for procurement of tissues and judicious selection of donors, microorganisms are frequently found in procured bones. Purpose of this study is to evaluate the factors that increase safety of procurement and minimize discard of procured tissues. Microbiological contamination was analyzed in 1271 musculoskeletal tissues removed from 138 multi-organ donors over a period extending from 2006 to 2016. Effects of various risk factors related with contamination were estimated using a logistic regression model. Microbiological contamination rate in the tissues was 17.1%; low pathogenic microorganisms were cultivated in 12.9% of the tissues, while highly pathogenic ones were cultivated in 4.2% of the tissues. Evolution of one single team was monitored during that period, verifying a fall in the general contamination level from 22.5 to 9.2%. Absence of antibiotics increased low pathogenic contamination risk. Every additional day in intensive care unit (ICU) increased the risk of highly pathogenic contamination. Time elapsed between death and the beginning of removal procedures was found to be relevant for both low pathogenic and highly pathogenic microorganisms. Among the studied factors, the following contributed for a significant increase in contamination by microorganisms in removed tissues: lack of use of prophylactic antibiotic therapy in donors, quantity of removed tissues, length of admission in ICU and the time elapsed between aortic clamping and beginning of the removal procedure.

Comparison of automated and conventional microbiological examination of donated human cardiac tissue in heart valve banking.

Most tissue banks use the conventional method; however, the automated method has advantages over the conventional method. The aim of this study was to compare the conventional and automated methods of culture in human cardiac tissue using an artificial contamination model. Myocardial samples were contaminated with sequential concentration (104 to 10-1 CFU/mL) with Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus and Candida albicans. Cultures were obtained from solution were the fragment was immersed and minced tissue, before and after the routine decellularization solution, with automated and conventional culture methods. Automated and conventional methods were compared and a p value ≤ 0.05 was considered significant. Staphylococcus aureus presented a significantly higher growth in the automated method, as well as faster than the conventional (p < 0.05). The positivity for growth in the automated method was higher in concentrated inoculum (> 102 CFU/mL) (p < 0.05). The growth in the automated method was significantly faster than conventional when inoculum concentration was above 103 CFU/mL. The automated culture method is faster than conventional method with a higher positivity in a contaminated model of myocardial and transport solution used in tissue banks.

Bioburden in transport solutions of human cardiovascular tissues: a comparative evaluation of direct inoculation and membrane filter technique.

All cardiac allograft tissues are under potential contamination, requiring a validated terminal sterilization process or a minimal bioburden. The bioburden calculation is important to determine the bacterial burden and further decontamination and disinfection strategies for the valve processing. The aim of this study was to determine the bioburden from transport solution (TS) of heart valves obtained from non-heart-beating and heart-beating donors in different culture methods. The bioburden from TS was determined in 20 hearts donated for valve allograft tissue using membrane filter (MF) and direct inoculation. Tryptic soy agar and Sabouraud plates were incubated and colonies were counted. Ninety-five percent of samples from this study were obtained from heart-beating donors. The warm ischemic time period for heart was 1.06 ± 0.74 h and the cold ischemic time period was 25.66 ± 11.16 h. The mean TS volume was 232.68 ± 96.67 mL (48.5-550 mL). From 20 samples directly inoculated on TSA agar plates, 2 (10%) were positive. However, when MF was used, from 20 samples in TSA, 13 (65%) were positive with a mean count of 1.36 ± 4.04 CFU/mL. In Sabouraud plates, the direct inoculation was positive in 5 samples (25%) with a mean count of 0.24 ± 0.56 CFU/mL. The use of MF increased the positivity to 50% (10 samples from a total of 20) with a mean of 0.28 ± 0.68 CFU/mL. The positivity was superior using MF in comparison with direct inoculation (p < 0.05). The bioburden of TS is low and MF is the technique of choice due to higher positivity.

[Skin and tissue bank: Operational model for the recovery and preservation of tissues and skin allografts]. / Banco de piel y tejidos: un modelo operativo para la recuperación y preservación de aloinjertos de piel y tejidos.

Tissue storage is a medical process that is in the regulation and homogenisation phase in the scientific world. The international standards require the need to ensure safety and efficacy of human allografts such as skin and other tissues. The activities of skin and tissues banks currently involve their recovery, processing, storage and distribution, which are positively correlated with technological and scientific advances present in current biomedical sciences. A description is presented of the operational model of Skin and Tissue Bank at INR as successful case for procurement, recovery and preservation of skin and tissues for therapeutic uses, with high safety and biological quality. The essential and standard guidelines are presented as keystones for a tissue recovery program based on scientific evidence, and within an ethical and legal framework, as well as to propose a model for complete overview of the donation of tissues and organ programs in Mexico. Finally, it concludes with essential proposals for improving the efficacy of transplantation of organs and tissue programs.

[The clinical use of cryopreserved human skin allografts for transplantation]. / El uso clínico de aloinjertos de piel humana criopreservados con fines de trasplante.

BACKGROUND: The biological recovery of human skin allografts is the gold standard for preservation in Skin Banks. However, there is no worldwide consensus about specific allocation criteria for preserved human skin allografts with living cells. A report is presented on the results of 5 years of experience of using human skin allografts in burned patient in the Skin and Tissue Bank at the "Instituto Nacional de Rehabilitacion" MATERIAL AND METHODS: The human skin allografts were obtained from multi-organ donors. processed and preserved at -80 °C for 12 months. Allocation criteria were performed according to blood type match, clinical history, and burned body surface. RESULTS: Up to now, the Skin and Tissue Bank at 'Instituto Nacional de Rehabilitacion" has processed and recovered 125,000 cm(2) of human skin allografts. It has performed 34 surgical implants on 21 burned patients. The average of burn body surface was 59.2%. More than two-thirds (67.7%) of recipients of skin allografts were matched of the same to type blood of the donor, and 66.6% survived after 126 days hospital stay. CONCLUSION: It is proposed to consider recipient's blood group as allocation criteria to assign tissue; and use human skin allografts on patiens affected with burns over 30% of body surface (according the "rule of the 9").