RESUMO
Extracellular vesicles (EVs) have been described as important mediators of cell communication, regulating several physiological processes, including tissue recovery and regeneration. In the kidneys, EVs derived from stem cells have been shown to support tissue recovery in diverse disease models and have been considered an interesting alternative to cell therapy. For this purpose, however, several challenges remain to be overcome, such as the requirement of a high number of EVs for human therapy and the need for optimization of techniques for their isolation and characterization. Moreover, the kidney's complexity and the pathological process to be treated require that EVs present a heterogeneous group of molecules to be delivered. In this review, we discuss the recent advances in the use of EVs as a therapeutic tool for kidney diseases. Moreover, we give an overview of the new technologies applied to improve EVs' efficacy, such as novel methods of EV production and isolation by means of bioreactors and microfluidics, bioengineering the EV content and the use of alternative cell sources, including kidney organoids, to support their transfer to clinical applications.
Assuntos
Injúria Renal Aguda/terapia , Terapia Baseada em Transplante de Células e Tecidos/métodos , Exossomos/metabolismo , Vesículas Extracelulares/metabolismo , Células-Tronco Mesenquimais/metabolismo , Insuficiência Renal Crônica/terapia , Bioengenharia/métodos , Técnicas de Cultura de Células/métodos , Exossomos/transplante , Vesículas Extracelulares/transplante , Humanos , Células-Tronco Mesenquimais/citologia , Tamanho da PartículaRESUMO
SUMMARY: Cell culture is an important tool in medical, odontological and biological research laboratories, supporting cell therapies and tissue bioengineering strategies. Gingival fibroblasts present structural function, being able to modulate their metabolic capacity, which is reflected in the tissue morphology. The possibility of culturing fibroblasts in vitro, in monolayer or on three-dimensional scaffolds, for subsequent transplants in vivo opens important perspectives for the periodontal surgical clinic. The objective of the present article is to present a method of obtaining and cultivating viable human gingival fibroblasts for in vitro research. Explants derived from periodontal surgical discards were used, grown in 25 cm2 bottles to obtain a primary cell culture. After observing the proliferation and growth of the fibroblasts that interconnected and formed a monolayer network, involving the periphery of the explants, it was possible to remove the explants, to make the passage and the new subcultures were obtained in a ratio of 1:1. After 7 days, the amount of viable cells was analyzed in triplicate, using the Neubauer chamber technique, in cell culture bottles of 25 mm2 (T25) and 75 mm2 (T75). Fibroblasts were described and subclassified morphologically. The results showed a growth pattern in both bottles, but with a larger number in bottles of 75 cm2. Cells with fibroblastic morphology were subclassified into reticular and fusiform, being predominant those with fusiform morphology. In conclusion, culture of explant of human gingival connective tissue is a viable method for obtaining gingival connective tissue cells suitable for laboratory tests in cell culture, aiming at obtaining constructs for gingival tissue engineering.
RESUMEN: El cultivo celular es una herramienta importante en los laboratorios de investigación médica, odontológica y biológica, que apoyan las terapias celulares y las estrategias de bioingeniería de tejidos. Los fibroblastos gingivales presentan una función estructural, pudiendo modular su capacidad metabólica, que se refleja en la morfología tisular. La posibilidad de cultivar fibroblastos in vitro, en monocapa o en andamios tridimensionales, para trasplantes posteriores in vivo abre perspectivas importantes para la clínica de cirugía periodontal. El objetivo del presente artículo es presentar un método para obtener y cultivar fibroblastos gingivales humanos viables para investigación in vitro. Se utilizaron explantes derivados de los descartes quirúrgicos periodontales, crecidos en frascos de 25 cm2 para obtener un cultivo de células primarias. Después de observar la proliferación y el crecimiento de los fibroblastos que se interconectaron y formaron una red de monocapa, que involucraba la periferia de los explantes, fue posible eliminar los explantes, hacer el pasaje y los nuevos subcultivos se obtuvieron en una proporción de 1:1. Después de 7 días, la cantidad de células viables se analizó por triplicado, utilizando la técnica de cámara de Neubauer, en botellas de cultivo celular de 25 mm2 (T25) y 75 mm2 (T75). Los fibroblastos fueron descritos y sub-clasificados morfológicamente. Los resultados mostraron un patrón de crecimiento en ambas botellas, pero con un número mayor en botellas de 75 cm2. Las células con morfología fibroblástica se subclasificaron en reticulares y fusiformes, predominando aquellas con morfología fusiforme. En conclusión, el cultivo de explante de tejido conectivo gingival humano es un método viable para obtener células de tejido conectivo gingival adecuadas para pruebas de laboratorio en cultivos celulares, con el objetivo de obtener construcciones para la ingeniería del tejido gingival.
Assuntos
Humanos , Células do Tecido Conjuntivo , Técnicas de Cultura de Células/métodos , Bioengenharia/métodos , Gengiva/citologia , Biologia Celular , FibroblastosRESUMO
Bioengineering, which studies the principles and design of biological systems, is a field that has inspired the development of several technologies that are currently in use. In this work, we use concepts from the fish lateral line sensing mechanism and apply them to seismic imaging processing. The lateral line is a sensory system composed of an integrated array of mechanical sensors spanning along the fish body. We compare the array of sensors along body fish with the seismic acquisition, which employs an array of equally spaced identical mechanical sensors to image the Earth's subsurface. In both situations, the mechanical sensors capture and process mechanical vibrations from the environment to produce useful information. We explore the strategy of using the low-pass and high-pass sensors schema of fish lateral line to improve the seismic technique. We use the full-wave inversion method to compare the conventional acquisition procedure of identical sensors with alternative sets of different sensors, which mimics the fish lateral line. Our results show that the alternate sensors arrangement surpasses the performance of the conventional acquisition method, using just half of the input information. The results point at an image processing technique that is computationally more efficient and economical than the usual seismic processing method.
Assuntos
Bioengenharia/métodos , Desastres/prevenção & controle , Terremotos , Monitoramento Ambiental/instrumentação , Mecanorreceptores/fisiologia , Animais , Monitoramento Ambiental/métodos , Desenho de Equipamento , Peixes/fisiologia , Geografia , Processamento de Imagem Assistida por Computador , Sistema da Linha Lateral/citologia , Sistema da Linha Lateral/fisiologia , Sistemas MicroeletromecânicosRESUMO
In recent years, the demand for naturally derived products has hiked with enormous pressure to propose or develop state-of-the-art strategies to meet sustainable circular economy challenges. Microalgae possess the flexibility to produce a variety of high-value products of industrial interests. From pigments such as phycobilins or lutein to phycotoxins and several polyunsaturated fatty acids (PUFAs), microalgae have the potential to become the primary producers for the pharmaceutical, food, and agronomical industries. Also, microalgae require minimal resources to grow due to their autotrophic nature or by consuming waste matter, while allowing for the extraction of several valuable side products such as hydrogen gas and biodiesel in a single process, following a biorefinery agenda. From a Mexican microalgae biodiversity perspective, more than 70 different local species have been characterized and isolated, whereas, only a minimal amount has been explored to produce commercially valuable products, thus ignoring their potential as a locally available resource. In this paper, we discuss the microalgae diversity present in Mexico with their current applications and potential, while expanding on their future applications in bioengineering along with other industrial sectors. In conclusion, the use of available microalgae to produce biochemically revenuable products currently represents an untapped potential that could lead to the solution of several problems through green technologies. As such, if the social, industrial and research communities collaborate to strive towards a greener economy by preserving the existing biodiversity and optimizing the use of the currently available resources, the enrichment of our society and the solution to several environmental problems could be attained.
Assuntos
Biodiversidade , Bioengenharia/métodos , Produtos Biológicos/química , Microalgas/química , Crescimento Sustentável , Biocombustíveis , Biomassa , MéxicoRESUMO
Intestinal and free-living protozoa, such as Giardia lamblia, express a dense coat of variant-specific surface proteins (VSPs) on trophozoites that protects the parasite inside the host's intestine. Here we show that VSPs not only are resistant to proteolytic digestion and extreme pH and temperatures but also stimulate host innate immune responses in a TLR-4 dependent manner. We show that these properties can be exploited to both protect and adjuvant vaccine antigens for oral administration. Chimeric Virus-like Particles (VLPs) decorated with VSPs and expressing model surface antigens, such as influenza virus hemagglutinin (HA) and neuraminidase (NA), are protected from degradation and activate antigen presenting cells in vitro. Orally administered VSP-pseudotyped VLPs, but not plain VLPs, generate robust immune responses that protect mice from influenza infection and HA-expressing tumors. This versatile vaccine platform has the attributes to meet the ultimate challenge of generating safe, stable and efficient oral vaccines.
Assuntos
Giardia lamblia/química , Vacinas contra Influenza/imunologia , Proteínas de Membrana/imunologia , Infecções por Orthomyxoviridae/prevenção & controle , Proteínas de Protozoários/imunologia , Vacinas de Partículas Semelhantes a Vírus/imunologia , Adjuvantes Imunológicos , Administração Oral , Animais , Apresentação de Antígeno/efeitos dos fármacos , Bioengenharia/métodos , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Células Dendríticas/virologia , Feminino , Expressão Gênica , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , Humanos , Imunidade Inata/efeitos dos fármacos , Vacinas contra Influenza/administração & dosagem , Vacinas contra Influenza/genética , Masculino , Proteínas de Membrana/genética , Camundongos , Camundongos Transgênicos , Neuraminidase/genética , Neuraminidase/imunologia , Infecções por Orthomyxoviridae/imunologia , Infecções por Orthomyxoviridae/virologia , Estabilidade Proteica , Proteínas de Protozoários/genética , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/imunologia , Trofozoítos/química , Vacinação , Vacinas de Partículas Semelhantes a Vírus/administração & dosagem , Vacinas de Partículas Semelhantes a Vírus/genéticaRESUMO
The present study evaluated 13 strains of yeast for ethanol and xylitol production from xylose. Among them, Spathaspora hagerdaliae UFMG-CM-Y303 produced ethanol yields (YP/S) of 0.25 g g- 1 and 0.39 g g- 1 under aerobic and microaerophilic conditions, respectively, from a mixture of glucose and xylose in flasks. A pH of 5.0 and an inoculum of 3.0 × 108 cells mL- 1r resulted in the highest ethanol yields. These conditions were tested in a bioreactor for fermenting a medium containing an enzymatic hydrolysate of sugarcane bagasse with 15.5 g L- 1 of glucose and 3 g L- 1 of xylose, and achieved a YP/S of 0.47 g g- 1, in relation to total available sugar. These results suggest that S. hagerdaliae UFMG-CM-Y303 has potential for use in second-generation ethanol studies.
Assuntos
Celulose/metabolismo , Etanol/química , Glucose/química , Saccharomycetales/metabolismo , Saccharum/metabolismo , Xilose/química , Bioengenharia/métodos , Biomassa , Reatores Biológicos , Meios de Cultura , Fermentação , Concentração de Íons de Hidrogênio , Cinética , Lignina/química , Saccharomyces cerevisiae/metabolismo , Xilitol/químicaRESUMO
Bioengineered tissues have become increasingly more sophisticated owing to recent advancements in the fields of biomaterials, microfabrication, microfluidics, genetic engineering, and stem cell and developmental biology. In the coming years, the ability to engineer artificial constructs that accurately mimic the compositional, architectural, and functional properties of human tissues, will profoundly impact the therapeutic and diagnostic aspects of the healthcare industry. In this regard, bioengineered cardiac tissues are of particular importance due to the extremely limited ability of the myocardium to self-regenerate, as well as the remarkably high mortality associated with cardiovascular diseases worldwide. As novel microphysiological systems make the transition from bench to bedside, their implementation in high throughput drug screening, personalized diagnostics, disease modeling, and targeted therapy validation will bring forth a paradigm shift in the clinical management of cardiovascular diseases. Here, we will review the current state of the art in experimental in vitro platforms for next generation diagnostics and therapy validation. We will describe recent advancements in the development of smart biomaterials, biofabrication techniques, and stem cell engineering, aimed at recapitulating cardiovascular function at the tissue- and organ levels. In addition, integrative and multidisciplinary approaches to engineer biomimetic cardiovascular constructs with unprecedented human and clinical relevance will be discussed. We will comment on the implementation of these platforms in high throughput drug screening, in vitro disease modeling and therapy validation. Lastly, future perspectives will be provided on how these biomimetic platforms will aid in the transition towards patient centered diagnostics, and the development of personalized targeted therapeutics.
Assuntos
Bioengenharia/instrumentação , Biomimética/instrumentação , Doenças Cardiovasculares/tratamento farmacológico , Doenças Cardiovasculares/patologia , Avaliação Pré-Clínica de Medicamentos/instrumentação , Animais , Materiais Biocompatíveis/química , Bioengenharia/métodos , Biomimética/métodos , Doenças Cardiovasculares/diagnóstico , Descoberta de Drogas/instrumentação , Descoberta de Drogas/métodos , Avaliação Pré-Clínica de Medicamentos/métodos , Desenho de Equipamento , Humanos , Dispositivos Lab-On-A-ChipRESUMO
Introdução: A expansão da pele é um processo fisiológico definido como a capacidade de aumentar sua área superficial em resposta a uma tensão ou a uma dada deformação. Para realizar a cirurgia reconstrutiva, os expansores de pele são implantados sob a pele e periodicamente infiltrados com uma solução salina para fornecer um retalho extra de pele. Quando o volume interno prescrito do expansor é alcançado, a cirurgia reconstrutiva é realizada. Métodos: Foi desenvolvido um dispositivo semiautomático e portátil para facilitar um procedimento de expansão da pele. O dispositivo tem como objetivo simplificar o processo de infiltração, proporcionando mobilidade e independência para o paciente, e assegurando ao médico a qualidade e a precisão das infiltrações realizadas. O dispositivo também permite expansão contínua em pacientes hospitalizados. Resultados: Usando um código, o médico tem acesso ao menu do dispositivo e define a pressão máxima e/ou o valor máximo para cada expansor do paciente. O paciente pode realizar a infiltração e ter acesso ao controle da velocidade de infiltração, reverter ou parar a operação. Todos os dados são gravados em um SIM Card e incluem data, hora, volumes inicial e final, e pressão inicial e final de cada procedimento para cada expansor. Conclusões: O dispositivo automatiza e otimiza a expansão, de modo que o que o médico possa prescrever um limite para cada expansão, seja uma pressão máxima ou voluma infiltrado. Todos os dados são gravados, fornecendo um importante banco de dados sobre o comportamento de pele relacionado a gênero, raça, idade e local da expansão.
Introduction: Skin expansion is a physiological process defined as the ability of human skin to increase its superficial area in response to a stress or given deformation. In reconstructive surgery, skin expanders are implanted beneath the skin and periodically infiltrated with a saline solution to provide an extra flap of skin. When the prescribed internal volume of the expander is reached, reconstructive surgery is performed. Methods: A semiautomatic and portable device was developed and built to facilitate a skin expansion procedure. The device aims to simplify the infiltration process, providing mobility and independence to the patient and assuring the physician of the infiltration quality and precision. The device also enables continuous expansion in hospitalized patients. Results: Using a code, the doctor accesses the menu of the device and sets the maximum pressure and/or value for each expander of the patient. The patient can control the infiltration velocity and reverse or stop the operation. All data are recorded on a simcard and include date, time, initial and final volumes, and initial and final pressures of each procedure for each expander. Conclusions: The device motorizes and optimizes the expansion, allowing the doctor to prescribe a maximum infiltration pressure or volume. All data are recorded to provide an important database of skin behavior related to sex, race, age, and expansion site.
Assuntos
Humanos , Expansão de Tecido/métodos , Procedimentos de Cirurgia Plástica/métodos , Bioengenharia/métodos , Procedimentos Cirúrgicos Ambulatórios/efeitos adversos , Procedimentos Cirúrgicos Ambulatórios/métodos , Avulsões Cutâneas/cirurgia , Avulsões Cutâneas/fisiopatologia , Dispositivos para Expansão de TecidosRESUMO
Solid-state fermentation (SSF) has been largely employed during the last three decades to produce different biomolecules of industrial interest, particularly enzymes. Through the use of agroindustrial wastes as SSF substrates, an economic process of lipases production can be achieved. In this chapter we describe a comprehensive SSF method for producing an economical preparation of Rhizomucor miehei lipase, employing sugarcane bagasse and used vegetal oil as substrates. To demonstrate the usefulness of the lipase produced by this method, we utilized directly the dried fermented solid, as a heterogeneous biocatalyst for the ethanolysis of different fats and oils. Final ethyl ester conversions (>90%, 24 h) were similar with those obtained using a commercial immobilized Rhizomucor miehei lipase at our best conditions. In this work we demonstrated that SSF is an easy and economical method for the production of lipases that can be used directly as heterogeneous biocatalysts for biodiesel production, employing low-cost feedstocks.
Assuntos
Bioengenharia , Fermentação , Lipase/biossíntese , Bioengenharia/instrumentação , Bioengenharia/métodos , Biocombustíveis , Catálise , Concentração de Íons de Hidrogênio , Hidrólise , Cinese , Lipase/isolamento & purificação , TemperaturaRESUMO
BACKGROUND: Despite its ability to grow and produce high-value molecules using renewable carbon sources, two main factors must be improved to use Burkholderia sacchari as a chassis for bioproduction at an industrial scale: first, the lack of molecular tools to engineer this organism and second, the inherently slow growth rate and poly-3-hydroxybutyrate [P(3HB)] production using xylose. In this work, we have addressed both factors. RESULTS: First, we adapted a set of BglBrick plasmids and showed tunable expression in B. sacchari. Finally, we assessed growth rate and P(3HB) production through overexpression of xylose transporters, catabolic or regulatory genes. Overexpression of xylR significantly improved growth rate (55.5% improvement), polymer yield (77.27% improvement), and resulted in 71% of cell dry weight as P(3HB). CONCLUSIONS: These values are unprecedented for P(3HB) accumulation using xylose as a sole carbon source and highlight the importance of precise expression control for improving utilization of hemicellulosic sugars in B. sacchari.