RESUMO
Biological biomaterials for tissue engineering purposes can be produced through tissue and/or organ decellularization. The remaining extracellular matrix (ECM) must be acellular and preserve its proteins and physical features. Placentas are organs of great interest because they are discarded after birth and present large amounts of ECM. Protocols for decellularization are tissue-specific and have not been established for canine placentas yet. This study aimed at analyzing a favorable method for decellularization of maternal and fetal portions of canine placentas. Canine placentas were subjected to ten preliminary tests to analyze the efficacy of parameters such as the type of detergents, freezing temperatures and perfusion. Two protocols were chosen for further analyses using histology, scanning electron microscopy, immunofluorescence and DNA quantification. Sodium dodecyl sulfate (SDS) was the most effective detergent for cell removal. Freezing placentas before decellularization required longer periods of incubation in different detergents. Both perfusion and immersion methods were capable of removing cells. Placentas decellularized using Protocol I (1% SDS, 5 mM EDTA, 50 mM TRIS, and 0.5% antibiotic) preserved the ECM structure better, but Protocol I was less efficient to remove cells and DNA content from the ECM than Protocol II (1% SDS, 5 mM EDTA, 0.05% trypsin, and 0.5% antibiotic).
Assuntos
Animais , Feminino , Gravidez , Cães , Placenta/citologia , Engenharia Tecidual/métodos , Matriz Extracelular , Feto/citologia , Dodecilsulfato de Sódio/farmacologia , Materiais Biocompatíveis , Microscopia Eletrônica de Varredura , Reprodutibilidade dos Testes , Imunofluorescência , Colágeno/análise , Fibronectinas/análise , Laminina/análise , Ácido Edético , Temperatura Baixa , Engenharia Tecidual/veterinária , ImersãoRESUMO
Biological biomaterials for tissue engineering purposes can be produced through tissue and/or organ decellularization. The remaining extracellular matrix (ECM) must be acellular and preserve its proteins and physical features. Placentas are organs of great interest because they are discarded after birth and present large amounts of ECM. Protocols for decellularization are tissue-specific and have not been established for canine placentas yet. This study aimed at analyzing a favorable method for decellularization of maternal and fetal portions of canine placentas. Canine placentas were subjected to ten preliminary tests to analyze the efficacy of parameters such as the type of detergents, freezing temperatures and perfusion. Two protocols were chosen for further analyses using histology, scanning electron microscopy, immunofluorescence and DNA quantification. Sodium dodecyl sulfate (SDS) was the most effective detergent for cell removal. Freezing placentas before decellularization required longer periods of incubation in different detergents. Both perfusion and immersion methods were capable of removing cells. Placentas decellularized using Protocol I (1% SDS, 5 mM EDTA, 50 mM TRIS, and 0.5% antibiotic) preserved the ECM structure better, but Protocol I was less efficient to remove cells and DNA content from the ECM than Protocol II (1% SDS, 5 mM EDTA, 0.05% trypsin, and 0.5% antibiotic).
Assuntos
Matriz Extracelular , Feto/citologia , Placenta/citologia , Engenharia Tecidual/métodos , Animais , Materiais Biocompatíveis , Temperatura Baixa , Colágeno/análise , Cães , Ácido Edético , Feminino , Fibronectinas/análise , Imunofluorescência , Imersão , Laminina/análise , Microscopia Eletrônica de Varredura , Gravidez , Reprodutibilidade dos Testes , Dodecilsulfato de Sódio/farmacologia , Tensoativos/farmacologia , Engenharia Tecidual/veterináriaRESUMO
Despite the medical importance of Paederus beetles, no studies have studied the influence of the abiotic factors on the flight activity and nighttime dispersal of these insects in Brazil. Therefore, the influence of both climatic factors and moon phase on black-light catches of Paederus rove beetles was investigated. Paederus beetles were attracted to a black light source hourly from 1800 to 0600 hours, and data on weather conditions as well as moon phase data were taken for every sampling date. Overall, 543 individuals of Paederus beetles belonging to four species were captured: P. protensus, P. columbinus, P. brasiliensis, and P. mutans. Paederus beetles were mostly active in the warmest parts of the studied nights. Variations in nighttime temperature, relative humidity, wind speed, cloud cover, and moon phases appear not to affect Paederus flight. The diurnal temperature was observed to affect the night hourly dispersal of Paederus rove beetles as well as their distribution pattern during the entire period of study. The true environmental condition responsible for Paederus beetles seasonal pattern and daily night dispersal in northeastern Brazil were the annual moisture and drought cycles and the diurnal maximum temperatures, respectively. Significant trap catches were observed in the earliest hours after sunset (1800-2100), and people must be aware of this fact, as it can notably increase the risk of acquiring linearis dermatitis from the contact with large numbers of active Paederus.