Challenges in the use of nanostructures as carriers of nucleic acids in clinical practice

ABSTRACT The delivery of nucleic acids to cells is considered a crucial step for the success of genetic modifications aimed at therapeutic purposes or production of genetically modified animals. In this context, nanotechnology is one of the most promising fields of science, with the potential to solve several existing problems. Nanostructures have desirable characteristics to be used as carriers, such as nanometric size, large surface area, cell internalization capacity, prolonged and controlled release, among others. Genetically modified animals can contribute to the production of biopharmaceuticals, through the expression of high-associated-value molecules. The production of these animals, also known as biofactories, further enhances Brazilian agribusiness, since it allows adding value to the final product, and favors the integration between the agricultural market and the pharmaceutical sector. However, there is a growing concern about the safety and possible harmful effects of nanostructures, since data on the safe use of these materials are still insufficient. The objective of this review was to address aspects of the use of nanostructures, mainly carbon nanotubes as nucleic acid carriers, aiming at the production of genetically modified animals, with the certainty that progress in this field of knowledge depends on more information on the mechanisms of interaction between nanostructures, cells and embryos, as well as on its toxicity.

Avanços tecnológicos da criopreservação de ovócitos / Technological advances in oocytes criopreservation

Apresenta-se uma revisão atualizada sobre a criopreservação de ovócito e sua importância em reprodução assistida. O presente trabalho tem como ênfase vantagens e desvantagens dos métodos utilizados para congelamento de gametas, efeito dos crioprotetores intracelulares e extracelulares, além da aplicabilidade da técnica na fertilidade feminina e na manutenção da biodiversidade animal.

It presents an updated review of oocyte cryopreservation and its importance in assisted reproduction. Th is work has an emphasis on advantages and disadvantages of the methods used for freezing gametes, the eff ects of intracellular and extracellular cryoprotectants, and the applicability of the technique in female fertility and in maintaining animal biodiversity

Embriões F1 (Hol X Gir) produzidos in vitro utilizados a fresco em rebanhos leiteiros comerciais e controlado / F1 Holstein (Friesan x Gir) embryos produced in vitro used fresh in commercial dairy herds and controlled herds

A importância do (F1) para o Brasil é notória, associando rusticidade e produtividade, porém é grande a dificuldade do produtor em manter seu rebanho nesse grau de sangue. No laboratório da Embrapa Gado de Leite em Juiz de Fora, MG foram produzidos in vitro 266 embriões (F1), a partir de complexos cumulus-oócitos (CCO's) de vacas Gir puncionadas e fecundadas com sêmen de touro Holandês. Destes, 53 embriões foram transferidos a fresco para receptoras de rebanhos comerciais de uma Cooperativa da Zona da Mata, MG, enquanto 72 embriões foram inovulados em rebanho controlado da Embrapa Campo Experimental de Coronel Pacheco. A produção de embriões F1 por fecundação in vitro apresentou taxa de produção de blastocisto de 15,58%. A taxa de gestação obtida foi de 26,4% no rebanho comercial e 40,2% no rebanho da Embrapa. Considerando-se o nascimento de 50% de machos e fêmeas, conclui-se que em rebanhos de 100 vacas seriam obtidas 13 fêmeas (13% de reposição) nos rebanhos comerciais e 20 fêmeas (20% de reposição) no rebanho da Embrapa. Foram muitas as dificuldades de operacionalização para o uso de dessa biotecnologia que requer um mínimo de escrituração zootécnica, controle sanitário, observação de cios e manejo reprodutivo.

The importance of (F1) to Brazil is notable, associating productivity and rusticity, it is difficult to the producer maintaining the herd in this blood degree. In Embrapa's Gado de Leite, laboratory in the city of Juiz de Fora, M.G., 266 embryos (F1) were produced in vitro from the oocytes of Gir cows, punched and fecundated with Holstein Friesan bull semen. Out of these, 53 embryos were transferred at natural into recipient cows in the commercial herds indicated by the technicians of a great cooperative of Zona da Mata's area, 72 embryos were transferred into recipients of Embrapa's herds under nutritional and sanitary controls. The remainder 139 embryos were frozen due to the lack of adequate recipient cows. The production of F1 embryos through in vitro fecundation of oocytes from Gir cows with semen from Holstein Friesan bull blastocist production rate of 15,58 %. The pregnance rate obtained was 26.4% in the cows from the commercial herds and 40.2% in the herds of Embrapa Gado de Leite. Considering the birth of 50% female cows, we can estimate a replacement annual rate of 13% (26.2% for gestation) for commercial herds and 20% (40% for pregnancy) for the controlled herd. There were many difficulties encountered in the operation, such as (i) the low level of adoption of technology; (ii) little investment in feeding, handling and sanity conditions of the herds; (iii) conditions capable to compromising the utilization of embryos, which requires a minimum of structure in terms of zoo-technical documentation, observation of rutting and reproduction management.