RESUMO
Pancreatic bioengineering is a potential therapeutic alternative for type 1 diabetes (T1D) in which the pancreas is decellularized, generating an acellular extracellular matrix (ECM) scaffold, which may be reconstituted by recellularization with several cell types to generate a bioartificial pancreas. No consensus for an ideal pancreatic decellularization protocol exists. Therefore, we aimed to determine the best-suited detergent by comparing sodium dodecyl sulfate (SDS), sodium deoxycholate (SDC), and Triton X-100 at different concentrations. Murine (n=12) and human pancreatic tissue from adult brain-dead donors (n=06) was harvested in accordance with Institutional Ethical Committee of the University of São Paulo Medical School (CEP-FMUSP) and decellularized under different detergent conditions. DNA content, histological analysis, and transmission and scanning electron microscopy were assessed. The most adequate condition for pancreatic decellularization was found to be 4% SDC, displaying: a) effective cell removal; b) maintenance of extracellular matrix architecture; c) proteoglycans, glycosaminoglycans (GAGs), and collagen fibers preservation. This protocol was extrapolated and successfully applied to human pancreas decellularization. The acellular ECM scaffold generated was recelullarized using human pancreatic islets primary clusters. 3D clusters were generated using 0.5×104 cells and then placed on top of acellular pancreatic slices (25 and 50 µm thickness). These clusters tended to connect to the acellular matrix, with visible cells located in the periphery of the clusters interacting with the ECM network of the bioscaffold slices and continued to produce insulin. This study provided evidence on how to improve and accelerate the pancreas decellularization process, while maintaining its architecture and extracellular structure, aiming at pancreatic bioengineering.
Assuntos
Ácido Desoxicólico , Detergentes , Pâncreas , Dodecilsulfato de Sódio , Engenharia Tecidual , Alicerces Teciduais , Animais , Detergentes/química , Detergentes/farmacologia , Humanos , Pâncreas/citologia , Camundongos , Dodecilsulfato de Sódio/farmacologia , Ácido Desoxicólico/farmacologia , Ácido Desoxicólico/química , Alicerces Teciduais/química , Engenharia Tecidual/métodos , Octoxinol/química , Matriz Extracelular , Diabetes Mellitus Tipo 1 , Microscopia Eletrônica de Varredura , Matriz Extracelular Descelularizada/químicaRESUMO
Pancreatic bioengineering is a potential therapeutic alternative for type 1 diabetes (T1D) in which the pancreas is decellularized, generating an acellular extracellular matrix (ECM) scaffold, which may be reconstituted by recellularization with several cell types to generate a bioartificial pancreas. No consensus for an ideal pancreatic decellularization protocol exists. Therefore, we aimed to determine the best-suited detergent by comparing sodium dodecyl sulfate (SDS), sodium deoxycholate (SDC), and Triton X-100 at different concentrations. Murine (n=12) and human pancreatic tissue from adult brain-dead donors (n=06) was harvested in accordance with Institutional Ethical Committee of the University of São Paulo Medical School (CEP-FMUSP) and decellularized under different detergent conditions. DNA content, histological analysis, and transmission and scanning electron microscopy were assessed. The most adequate condition for pancreatic decellularization was found to be 4% SDC, displaying: a) effective cell removal; b) maintenance of extracellular matrix architecture; c) proteoglycans, glycosaminoglycans (GAGs), and collagen fibers preservation. This protocol was extrapolated and successfully applied to human pancreas decellularization. The acellular ECM scaffold generated was recelullarized using human pancreatic islets primary clusters. 3D clusters were generated using 0.5×104 cells and then placed on top of acellular pancreatic slices (25 and 50 μm thickness). These clusters tended to connect to the acellular matrix, with visible cells located in the periphery of the clusters interacting with the ECM network of the bioscaffold slices and continued to produce insulin. This study provided evidence on how to improve and accelerate the pancreas decellularization process, while maintaining its architecture and extracellular structure, aiming at pancreatic bioengineering.
RESUMO
We report on a statistical analysis of the engagement in the electoral processes of all Brazilian cities by considering the number of party memberships and the number of candidates for mayor and councillor. By investigating the relationships between the number of party members and the population of voters, we have found that the functional forms of these relationships are well described by sublinear power laws (allometric scaling) surrounded by a multiplicative log-normal noise. We have observed that this pattern is quite similar to those we previously reported for the relationships between the number of candidates (mayor and councillor) and population of voters [Europhys. Lett. 96, 48001 (2011)], suggesting that similar universal laws may be ruling the engagement in the electoral processes. We also note that the power-law exponents display a clear hierarchy, where the more influential is the political position the smaller is the value of the exponent. We have also investigated the probability distributions of the number of candidates (mayor and councillor), party memberships, and voters. The results indicate that the most influential positions are characterized by distributions with very short tails, while less influential positions display an intermediate power-law decay before showing an exponential-like cutoff. We discuss the possibility that, in addition to the political power of the position, limitations in the number of available seats can also be connected with this changing of behavior. We further believe that our empirical findings point out to an under-representation effect, where the larger the city is, the larger are the obstacles for more individuals to become directly engaged in the electoral process.