Ciencia e Ingeniería en Alimentos y Biotecnología
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Item Elaboración de andamios biológicos mediante lixiviación de microesferas para aplicaciones biomédicas(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2024-08) Freire Galarza, Melissa Belen; Núñez Villacís, Lorena de los ÁngelesBone defects are one of the main global health problems, and the scarcity of conventional substitutes to treat this deficiency is a clinical disadvantage. Thus, bone tissue engineering (BTE) through the combination of cells, growth factors and scaffolds has emerged as a promising alternative in the regeneration of functional tissues. In this study, chitosan and polycaprolactone (PCL) biological scaffolds (ABs) were prepared by leaching gelatin microspheres at different concentrations of 25, 50 and 75 percent in order to evaluate their impact on morphological, mechanical and biological properties. Macroporous and microporous scaffolds (90 to 600 micrometers) were obtained, the scaffolds with a higher concentration of microspheres presented a porosity of 85.29 percent, proving to be more favorable on the other properties of the ABs, with a water vapor transmission rate of 117. 18 grams per square meter per-24 hours, a water absorption capacity of 249.02 percent and a biodegradability of 31.09 and 54.41 percent in simulated body fluid (SBF), and blood plasma respectively, differing only in mechanical properties with a modulus of elasticity of 386.70 kilopascals, which indicated a lower stiffness of the material. The resulting ABs demonstrated suitable properties that can be applied in ITO, depending on the characteristics, scaffolds with higher porosity for spongy bones and rapid cell integration, and scaffolds with lower porosity for dense bones requiring higher mechanical strength.Item Extracción de nanocelulosa vegetal a partir de residuos de banano (Musa paradisiaca) y su aplicación en andamios biológicos(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2024-02) Núñez Martínez, Kimberlyn Carolina; Núñez Villacís, Lorena de los ÁngelesThe wound healing process is highly complex and alterations in its pathway can cause severe scenarios with high economic cost. Tissue Engineering (TE) combines cells, scaffolds, and growth factors to give rise to new tissue. Scaffolds function as a support for cell growth and are fabricated with synthetic or natural biopolymers. Nanocellulose is a natural polymer that can be obtained from renewable sources, such as banana waste. This study focused on the extraction of nanocellulose from banana waste and its subsequent application in the fabrication of biological scaffolds for future research in the healing of chronic wounds. Banana nanocellulose was extracted by hydrolysis with sulfuric acid and ultrasound. Its morphology was evaluated by inverted light microscopy and SEM. In addition, the main functional groups present in nanocellulose were analyzed by FTIR. Subsequently, nanocellulose, chitosan, and nanocellulose whit chitosan scaffolds were prepared by freeze-drying in order to evaluate the characteristics of nanocellulose versus other biopolymers. Scaffolds were then evaluated by SEM, porosity tests, water absorption capacity and biodegradability tests. A yield of 23.8 percent of extracted nanocellulose was obtained. The scaffolds composite of nanocellulose and chitosan showed the best results with an average pore size of 110.13 micrometers with 88.94 percent. porosity, 2418.32 percent of water adsorption and an ability to degrade in 16 days. The results, suggest that these scaffold have potential to be applied in TE research related to wound healing and tissue regeneration.Item Evaluación de la actividad antioxidante de extractos de Aloysia citrodora (Cedrón) y Prunus serotina (Capulí) en Andamios biológicos(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2024-02) Gavilanes Suárez, Jonathan André; Núñez Villacís, Lorena de los ÁngelesIschemic heart disease (IHD) is a disease caused by obstruction of blood flow, due to factors such as oxidative stress due to reactive oxygen species (ROS). In certain cases, tissue transplants are required, so tissue engineering (TE) proposes possible alternatives for their treatment, such as the application of biological scaffolds (BSs) with antioxidants. This study focused on the evaluation of the antioxidant activity of extracts of Aloysia citrodora (cedrón) and Prunus serotina (capulí) in BSs. Extracts were obtained by maceration and the content of phenols and flavonoids was quantified. By inhibiting DPPH, its antioxidant activity was calculated and a combination of extracts was made. The best combination of extracts was incorporated in the preparation of BSs. To check the antioxidant activity in BS, FT-IR was performed in addition to a DPPH test. Morphological and physical properties were also evaluated by SEM, mechanical tests and permeability. The yield percentages in obtaining extracts were 11.34 for lemon verbena and 12.32 for capulí. The combination of extracts in a 7:3 ratio for capulí:cedron presented an increase in the percentage of inhibition of free radicals compared to pure extracts. The addition of the extract in ABs increased the inhibition percentage to 70.17. Its incorporation into BSs allows the presence of ROS to be reduced. This shows that BS with extract has the potential to be applied in TE for the development of endothelial tissue and combat IHD.Item Caracterización in vitro de andamios biológicos elaborados a partir de biopolímeros naturales para aplicaciones en ingeniería de tejidos(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2023-09) Ramírez Manzano, Víctor Xavier; Núñez Villacís, Lorena de los ÁngelesThe shortage of organ donors and the lack of techniques to regenerate injured tissues pose a challenge worldwide, making tissue engineering a promising alternative. In this discipline, cells, biological scaffolds, and factors are combined to generate functional tissues. This study focused on the development and characterization of scaffolds made from natural biopolymers, as these provide the basis for cell adhesion and proliferation. Collagen, chitosan, and collagen - chitosan (hybrid) scaffolds were prepared by lyophilization. Their morphology was evaluated by SEM (porosity) and ethanol infiltration (porosity percentage). In addition, hemocompatibility was characterized by swelling, clotting index, hemolysis, and red blood cell and platelet adhesion tests. The results showed that the hybrid scaffold has the best properties and is the most suitable for future research. However, it is proposed to use the collagen scaffold for epithelial tissue studies, the chitosan scaffold for bone and cartilage tissues, and the hybrid scaffold for bone, cartilage, and connective tissues. Future studies will evaluate the bioactivity of the scaffolds with cells, therefore, to ensure sterility of the scaffolds for cell culture, two sterilization methods were investigated: ultraviolet irradiation and ethanol (65-70 percent). UV light proved to be the best method due to the time required and ease of use. In conclusion, the characterization carried out on the ABs suggests that they possess adequate characteristics for future investigations related to tissue development; however, the collagen - chitosan scaffold presents better morphological characteristics. These ABs will be used in future evaluations of cell growth and proliferation.Item Caracterización in vitro de andamios biológicos elaborados a partir de biopolímeros naturales para aplicaciones en ingeniería de tejidos(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2023-09) Piñaloza Vásconez, Leslie de los Ángeles; Núñez Villacís, Lorena de los ÁngelesThe shortage of organ donors and the lack of techniques to regenerate injured tissues pose a challenge worldwide, making tissue engineering a promising alternative. In this discipline, cells, biological scaffolds, and factors are combined to generate functional tissues. This study focused on the development and characterization of scaffolds made from natural biopolymers, as these provide the basis for cell adhesion and proliferation. Collagen, chitosan, and collagen - chitosan (hybrid) scaffolds were prepared by lyophilization. Their morphology was evaluated by SEM (porosity) and ethanol infiltration (porosity percentage). In addition, hemocompatibility was characterized by swelling, clotting index, hemolysis, and red blood cell and platelet adhesion tests. The results showed that the hybrid scaffold has the best properties and is the most suitable for future research. However, it is proposed to use the collagen scaffold for epithelial tissue studies, the chitosan scaffold for bone and cartilage tissues, and the hybrid scaffold for bone, cartilage, and connective tissues. Future studies will evaluate the bioactivity of the scaffolds with cells, therefore, to ensure sterility of the scaffolds for cell culture, two sterilization methods were investigated: ultraviolet irradiation and ethanol (65-70 percent). UV light proved to be the best method due to the time required and ease of use. In conclusion, the characterization carried out on the ABs suggests that they possess adequate characteristics for future investigations related to tissue development; however, the collagen - chitosan scaffold presents better morphological characteristics. These ABs will be used in future evaluations of cell growth and proliferation.Item Elaboración de andamios biológicos a partir de polímeros naturales recubiertos con extracto de Clinopodium tomentosum (Kunth)(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2023-09) Carrera Fonseca, Geovanna Noemí; Tubón Usca, Irvin RicardoThe skin, being the largest organ in the body, is highly susceptible to damage from accidents and injuries, leading to tissue compromise. In certain cases, cell regeneration necessitates transplantation. In this context, tissue engineering and advancements in regenerative medicine have explored the potential of biological scaffolds. However, in Ecuador, the use of herbal extracts is limited due to unexplored knowledge and underutilization of the country's rich diversity of plant species. Among these valuable resources lies Clinopodium tomentosum (Kunth), an endemic species with antioxidant and angiogenic properties, holding promise for wound healing applications. The focus of this study was on the fabrication of scaffolds containing C. tomentosum (Kunth) extract at concentrations of 50 and 100 micrograms per milliliter, with collagen-chitosan hybrid scaffolds serving as controls. The extract underwent qualitative and microbiological tests to ensure its quality. The lyophilization technique preserved the 3D structure of the scaffolds while retaining the secondary metabolites of the extract, a finding corroborated through scanning electron microscopy images. Comprehensive evaluations of morphological, physical, and biological properties were conducted. Among the tested scaffolds, the one with 50 micrograms per milliliter of extract exhibited the most promising characteristics in terms of percentage and pore size. Moreover, it demonstrated favorable mechanical properties that ensure support after transplantation. Additionally, its biodegradability and antimicrobial properties against S. aureus make it a compelling candidate for application in epithelial wound healing.Item Obtención y caracterización de celulosa bacteriana producida por cepas acéticas endémicas del Ecuador(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2023-09) Álvarez Morales, Juan José; García Solís, Mario DanielThe research and development of new bacterial strains producing bacterial cellulose (BC) is a topic of great relevance today, especially in the context of biotechnology and industrial applications. This study focuses on native strains from Ecuador, with special attention to acetic bacteria PDC 21 and PDC 25, opening new possibilities for BC production. The methodology of this study involved the axenic culture of the strains and the observation of the formation of BC biofilm. Fermentative processes were carried out and the BC production yield was evaluated in comparison with a control of Komagataeibacter xylinus (DSMZ 2004). In addition, acid, thermal, and mechanical treatments were performed to obtain nanocellulose. The morphology of the samples was analyzed by scanning electron microscopy (SEM), while Fourier Transform Infrared Spectroscopy (FTIR-IR) was used to evaluate their chemical properties. The study concludes with the proposal that strain PDC 25 could be a viable alternative for BC production in industrial applications. This conclusion is based on its production yield, glucose consumption, and the properties of the BC it produces, which are similar to those of strain DSMZ 2004, commonly used as a study model and in the industry. This finding opens new possibilities for BC production and biotechnology in Ecuador.Item Elaboración de andamios biológicos a partir de biopolímeros naturales mediante métodos manuales y bioimpresión 3D para aplicaciones en ingeniería de tejidos(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2022-09) Guáitara Martínez, Sandro Daniel; Núñez Villacís, Lorena de los ÁngelesThere are thousands of patients with injuries that cannot be adequately treated, making tissue engineering a great alternative. Although it is not limited only to this because it can also be suitable for the development of drugs and the study of diseases. The aim of this research is to elaborate different scaffolds with potential applications in tissue engineering, using natural polymers. For this purpose, freeze-drying as a manual method and 3D bioprinting were used. The solutions for the scaffolds were elaborated by dissolving chitosan and collagen in acetic acid and mixing them with an agarose solution in PBS; these solutions were freeze-dried to obtain dry and porous scaffolds. For the 3D bioprinter, the solutions were printed in the form of 4 cm square squares, with a mesh inside. They were evaluated by SEM in order to measure the pore size and analyze their surface area. Using the ethanol infiltration method, a better understanding of the pores was obtained. Water absorption capacity tests were performed. Due to the importance of biodegradability, it was evaluated by immersion in blood plasma for 2, 4 and 7 days. Finally, the biomineralization of the scaffolds was also evaluated in the same way as biodegradability, i.e., incubating in blood plasma. With the evaluation of the characteristics, it was analyzed for which tissues the scaffolds could be useful, which was mainly bone and dermal tissue.Item Determinación de un método eficiente para la germinación in vitro de semillas de Zapoteca aculeata(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2022-09) Yugcha Yugcha, Michelle Estefanía; Vargas López, José HomeroEndemic species are often among the most sensitive to environmental changes and disturbances and therefore most at risk of extinction. To solve this problem, in vitro seed germination is a suitable method to propagate threatened plant species for conservation. The present work aimed to determine an efficient method for in vitro germination of Zapoteca aculeata seeds. First, two disinfection treatments were applied to the seeds to calculate the percentage of contamination. Then, six pre-germination treatments were applied to the seeds to evaluate the percentage and time of germination. Finally, the seed germination process was characterised. At each stage, the treatment that gave the best germination percentage was chosen using analysis of variance to statistically verify the significant difference of the treatments and the significant differences between the evaluated groups were evaluated using Tukey's test. The best results were obtained with disinfection D1, reaching a germination percentage of 27.08 percent, this is due to the concentration and exposure time of the seeds in the disinfectant solutions. With the pre-germinative method T2 a germination percentage of 46.25 percent was obtained, and the seeds were the earliest to germinate, taking between 17.4 days. Therefore, it is recommended to use it in future research because there is no information in the scientific literature regarding research work on the germination of Z. aculeata seeds.Item Aplicación de la ingeniería de tejidos en el desarrollo de válvulas cardiacas y miocardio(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería Bioquímica, 2022-03) Coronel Dávila, Karen Nicole; Núñez Villacís, Lorena de los ÁngelesHeart transplants are becoming more common medical procedure due to an increase in myocardial diseases. Unfortunately, the current number of myocardial donors is low and science is looking for other alternatives to obtain this organ or parts of it. The aim of this research was to analyze the application of tissue engineering for the development of heart valves and myocardium, as an alternative to the limited number of donors. This research analyzed the incidence of heart problems worldwide, obtaining a higher percentage of heart diseases in the Caucasian ethnic group. In addition, eight causes that influence the low number of available myocardial donors were identified, of which incompatibility and social perceptions were the most featured. Likewise, the problems that can occur when performing myocardial transplants were analyzed and it was found that infections are the most common problem. Subsequently, the alternatives that tissue engineering is developing to help overcoming these problems were investigated and compared. Currently six methods have been described that could be used as they have the advantage of using different types of cells and biomaterials that are adapted to individual's system. When comparing these techniques for both myocardium and heart valve development, it was found that cellular cardiomyoplasty is one of the best options to replace transplants due to its low cost, good functionality and easy implementation. In conclusion, tissue engineering is opening new doors for the development of complex organs and thus saving more lives.