RESUMEN
INTRODUCTION: Contradictory claims about the efficacy of several medicinal plants to promote glycemic control in patients with type 2 diabetes mellitus (T2DM) have been explained by divergences in the administration form and by extrapolation of data obtained from healthy individuals. It is not known whether the antidiabetic effects of traditional herbal medicines are influenced by gelatin capsules. This randomized crossover trial aimed to evaluate the acute effect of a single dose of raw cinnamon consumed orally either dissolved in water as a beverage or as ordinary hard gelatin capsules on postprandial hyperglycemia (>140 mg/dL; >7.8 mmol/L) in T2DM patients elicited by a nutritionally-balanced meal providing 50 g of complex carbohydrates. METHODS: Fasting T2DM patients (n = 19) randomly ingested a standardized meal in five experimental sessions, one alone (Control) and the other after prior intake of 3 or 6 g of crude cinnamon in the form of hard gelatin capsules or powder dissolved in water. Blood glucose was measured at fasting and at 0.25, 0.5, 0.75, 1, 1.5 and 2 hours postprandially. After each breakfast, its palatability scores for visual appeal, smell and pleasantness of taste were assessed, as well as the taste intensity sweetness, saltiness, bitterness, sourness and creaminess. RESULTS: The intake of raw cinnamon dissolved in water, independently of the dose, decreased the meal-induced large glucose spike (peak-rise of +87 mg/dL and Δ1-hour glycemia of +79 mg/dL) and the hyperglycemic blood glucose peak. When cinnamon was taken as capsules, these anti-hyperglycemic effects were lost or significantly diminished. Raw cinnamon intake did not change time-to-peak or the 2-h post-meal glycaemia, but flattened the glycemic curve (lower iAUC) without changing the shape that is typical of T2DM patients. CONCLUSIONS: This cinnamon's antihyperglycemic action confirms its acarbose-like property to inhibit the activities of the carbohydrate-digesting enzymes α-amylases/α-glucosidases, which is in accordance with its exceptionally high content of raw insoluble fiber. The efficacy of using raw cinnamon as a diabetes treatment strategy seems to require its intake at a specific time before/concomitantly the main hyperglycemic daily meals. Trial registration: Registro Brasileiro de Ensaios Clínicos (ReBEC), number RBR-98tx28b.
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Glucemia , Cápsulas , Estudios Cruzados , Diabetes Mellitus Tipo 2 , Gelatina , Hiperglucemia , Plantas Medicinales , Periodo Posprandial , Polvos , Humanos , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Diabetes Mellitus Tipo 2/sangre , Femenino , Masculino , Persona de Mediana Edad , Hiperglucemia/tratamiento farmacológico , Periodo Posprandial/efectos de los fármacos , Glucemia/análisis , Glucemia/efectos de los fármacos , Plantas Medicinales/química , Anciano , Cinnamomum zeylanicum/química , Adulto , Hipoglucemiantes/uso terapéutico , Hipoglucemiantes/administración & dosificaciónRESUMEN
In disease treatment, maintaining therapeutic drug concentrations often requires multiple doses. Lipid/polymer hybrid nanoparticles (LPHNPs) offer a promising solution by facilitating sustained drug delivery within therapeutic ranges. Here, we synthesized poly(lactic-co-glycolic acid) (PLGA) nanoparticles coated with soy lecithin using nanoprecipitation and self-assembly techniques. These nanoparticles were incorporated into gelatin aerogels to ensure uniform distribution and increase the concentration. Our study focused on understanding the release kinetics of hydrophilic (gallic acid) and lipophilic (quercetin) compounds from this system. Nanoparticles exhibited hydrodynamic diameters of 100 ± 15 nm (empty), 153 ± 33 nm (gallic acid-loaded), and 149 ± 21 nm (quercetin-loaded), with encapsulation efficiencies of 90 ± 5% and 70 ± 10% respectively. Gallic acid release followed the Korsmeyer-Peppas kinetics model (n = 1.01), while quercetin showed first-order kinetics. Notably, encapsulated compounds demonstrated delayed release compared to free compounds in gelatin aerogels, illustrating LPHNPs' ability to modulate release profiles independent of the compound type. This study underscores the potential of LPHNPs in optimizing drug delivery strategies for enhanced therapeutic outcomes.
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Ácido Gálico , Interacciones Hidrofóbicas e Hidrofílicas , Nanopartículas , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Quercetina , Quercetina/química , Nanopartículas/química , Ácido Gálico/química , Cinética , Copolímero de Ácido Poliláctico-Ácido Poliglicólico/química , Lecitinas/química , Gelatina/química , Ácido Láctico/química , Ácido Poliglicólico/química , Liberación de Fármacos , Lípidos/química , Portadores de Fármacos/química , Tamaño de la PartículaRESUMEN
The objective of this study was to create injectable photo-crosslinkable biomaterials, using gelatin methacryloyl (GelMA) hydrogel, combined with a decellularized bone matrix (BMdc) and a deproteinized (BMdp) bovine bone matrix. These were intended to serve as bioactive scaffolds for dentin regeneration. The parameters for GelMA hydrogel fabrication were initially selected, followed by the incorporation of BMdc and BMdp at a 1% (w/v) ratio. Nano-hydroxyapatite (nHA) was also included as a control. A physicochemical characterization was conducted, with FTIR analysis indicating that the mineral phase was complexed with GelMA, and BMdc was chemically bonded to the amide groups of gelatin. The porous structure was preserved post-BMdc incorporation, with bone particles incorporated alongside the pores. Conversely, the mineral phase was situated inside the pore opening, affecting the degree of porosity. The mineral phase did not modify the degradability of GelMA, even under conditions of type I collagenase-mediated enzymatic challenge, allowing hydrogel injection and increased mechanical strength. Subsequently, human dental pulp cells (HDPCs) were seeded onto the hydrogels. The cells remained viable and proliferative, irrespective of the GelMA composition. All mineral phases resulted in a significant increase in alkaline phosphatase activity and mineralized matrix deposition. However, GelMA-BMdc exhibited higher cell expression values, significantly surpassing those of all other formulations. In conclusion, our results showed that GelMA-BMdc produced a porous and stable hydrogel, capable of enhancing odontoblastic differentiation and mineral deposition when in contact with HDPCs, thereby showing potential for dentin regeneration.
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Pulpa Dental , Dentina , Gelatina , Ingeniería de Tejidos , Dentina/química , Ingeniería de Tejidos/métodos , Animales , Bovinos , Gelatina/química , Humanos , Pulpa Dental/citología , Metacrilatos/química , Reactivos de Enlaces Cruzados/química , Hidrogeles/química , Andamios del Tejido/química , Huesos , Células Cultivadas , PorosidadRESUMEN
Gelatin, a versatile protein derived from collagen, is widely used in the food, pharmaceutical and medical sectors. However, bacterial contamination by spore-forming bacteria during gelatin processing represents a significant concern for product safety and quality. In this study, an investigation was carried out to explore the heat and chemical resistance, as well as the identification and characterization of spore-forming bacteria isolated from gelatin processing. The methodologies involved chemical resistance tests with drastic pH in microplates and thermal resistance tests in capillary tubes of various isolates obtained at different processing stages. In addition, phenotypic and genotypic analyses were carried out to characterize the most resistant isolates of spore-forming bacteria. The findings of this study revealed the presence of several species, including Bacillus cereus, Bacillus licheniformis, Bacillus sonorensis, Bacillus subtilis, Geobacillus stearothermophilus, and Clostridium sporogenes, with some isolates exhibiting remarkable chemical and heat resistances. In addition, a significant proportion of the most resistant isolates showed gelatinase activity (n = 19/21; 90.5 %) and the presence of heat resistance (n = 5/21; 23.8 %), and virulence genes (n = 11/21; 52.4 %). The results of this study suggest that interventions should be done in quality control practices and that process parameter adjustments and effective contamination reduction strategies should be implemented through gelatin processing.
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Gelatina , Calor , ARN Ribosómico 16S , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Esporas Bacterianas , Esporas Bacterianas/genética , ARN Ribosómico 16S/genética , Virulencia/genética , Microbiología de Alimentos , Bacillus/genética , Bacillus/aislamiento & purificaciónRESUMEN
Scaffolds for the filling and regeneration of osteochondral defects are a current challenge in the biomaterials field, and solutions with greater functionality are still being sought. The novel approach of this work was to obtain scaffolds with biologically active additives possessing microstructural, permeability, and mechanical properties, mimicking the complexity of natural cartilage. Four types of scaffolds with a gelatin/alginate matrix modified with hydroxyapatite were obtained, and the relationship between the modifiers and substrate properties was evaluated. They differed in the type of second modifier used, which was hydrated MgCl2 in two proportions, ZnO, and nanohydroxyapatite. The samples were obtained by freeze-drying by using two-stage freezing. Based on microstructural observations combined with X-ray microanalysis, the microstructure of the samples and the elemental content were assessed. Permeability and mechanical tests were also performed. The scaffolds exhibited a network of interconnected pores and complex microarchitecture, with lower porosity at the surface (15 ± 7 to 29 ± 6%) and higher porosity at the center (67 ± 8 to 75 ± 8%). The additives had varying effects on the pore sizes and permeabilities of the samples. ZnO yielded the most permeable scaffolds (5.92 × 10-11 m2), whereas nanohydroxyapatite yielded the scaffold with the lowest permeability (1.18 × 10-11 m2), values within the range reported for trabecular bone. The magnesium content had no statistically significant effect on the permeability. The best mechanical parameters were obtained for ZnO samples and those containing hydrated MgCl2. The scaffold's properties meet the criteria for filling osteochondral defects. The developed scaffolds follow a biomimetic approach in terms of hierarchical microarchitecture and mechanical parameters as well as chemical composition. The obtained composite materials have the potential as biomimetic scaffolds for the regeneration of osteochondral defects.
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Hidrogeles , Cloruro de Magnesio , Andamios del Tejido , Óxido de Zinc , Óxido de Zinc/química , Andamios del Tejido/química , Cloruro de Magnesio/química , Hidrogeles/química , Porosidad , Alginatos/química , Durapatita/química , Permeabilidad , Gelatina/química , Ensayo de MaterialesRESUMEN
BACKGROUND: Polymeric electrospun mats have been used as scaffolds in tissue engineering for the development of novel materials due to its characteristics. The usage of synthetic materials has gone in decline due to environmental problems associated with their synthesis and waste disposal. Biomaterials such as biopolymers have been used recently due to good compatibility on biological applications and sustainability. OBJECTIVE: The purpose of this work is to obtain novel materials based on synthetic and natural polymers for applications on tissue engineering. METHODS: Aloe vera mucilage was obtained, chemically characterized, and used as an active compound contained in electrospun mats. Polymeric scaffolds were obtained in single, coaxial and tri-layer structures, characterized and evaluated in cell culture. RESULTS: Mucilage loaded electrospun fibers showed good compatibility due to formation of hydrogen bonds between polymers and biomolecules from its structure, evidenced by FTIR spectra and thermal properties. Cell viability test showed that most of the obtained mats result on viability higher than 75%, resulting in nontoxic materials, ready to be used on scaffolding applications. CONCLUSION: Mucilage containing fibers resulted on materials with potential use on scaffolding applications due to their mechanical performance and cell viability results.
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Aloe , Supervivencia Celular , Gelatina , Mucílago de Planta , Poliésteres , Ingeniería de Tejidos , Andamios del Tejido , Poliésteres/química , Ingeniería de Tejidos/métodos , Gelatina/química , Andamios del Tejido/química , Supervivencia Celular/efectos de los fármacos , Aloe/química , Mucílago de Planta/química , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Ensayo de Materiales , Humanos , Membranas Artificiales , AnimalesRESUMEN
Background: Desserts with vegetable ingredients are a constantly expanding global market due to the search for alternatives to cow's milk. Fermentation of these matrices by lactic acid bacteria can add greater functionality to the product, improving its nutritional, sensory, and food safety characteristics, as well as creating bioactive components with beneficial effects on health. Concern for health and well-being has aroused interest in byproducts of the industry that have functional properties for the body, such as mature coconut water, a normally discarded residue that is rich in nutrients. This study aimed to develop a probiotic gelatin based on pulp and water from mature coconuts and evaluate the physicochemical characteristics, viability of the Lacticaseibacillus rhamnosus LR32 strain in the medium, as well as the texture properties of the product. Methods: After collection and cleaning, the physicochemical characterization, mineral analysis, analysis of the total phenolic content and antioxidant activity of mature coconut water were carried out, as well as the centesimal composition of its pulp. Afterwards, the gelling was developed with the addition of modified corn starch, gelatin, sucrose, and probiotic culture, being subjected to acidity analysis, texture profile and cell count, on the first day and every 7 days during 21 days of storage, under refrigeration at 5 °C. An analysis of the centesimal composition was also carried out. Results: The main minerals in coconut water were potassium (1,932.57 mg L-1), sodium (19.57 mg L-1), magnesium (85.13 mg L-1) calcium (279.93 mg L-1) and phosphorus (11.17 mg L- 1), while the pulp had potassium (35.96 g kg-1), sodium (0.97 g kg-1), magnesium (2.18 g kg-1), 37 calcium (1.64 g kg-1), and phosphorus (3.32 g kg-1). The phenolic content of the water and pulp was 5.72 and 9.77 mg gallic acid equivalent (GAE) 100 g-1, respectively, and the antioxidant capacity was 1.67 and 0.98 39 g of 2, 2-diphenyl-1-picrylhydrazyl (DPPH) mg-1, respectively. The coconut pulp had 2.81 g 100 g-1of protein, 1.11 g 100 g-1 of 40 ash, 53% moisture, and 5.81 g 100 g-1 of carbohydrates. The gelatin produced during the storage period presented firmness parameters ranging from 145.82 to 206.81 grams-force (gf), adhesiveness from 692.85 to 1,028.63 gf sec, cohesiveness from 0.604 to 0.473, elasticity from 0.901 to 0.881, gumminess from 86.27 to 97.87 gf, and chewiness from 77.72 to 91.98 gf. Regarding the viability of the probiotic microorganism, the dessert had 7.49 log CFU g-1 that remained viable during the 21-day storage, reaching 8.51 CFU g-1. Acidity ranged from 0.15 to 0.64 g of lactic acid 100 g-1. The centesimal composition of the product showed 4.88 g 100 g-1 of protein, 0.54 g 100 g-1 of ash, 85.21% moisture, and 5.37g 100 g-1 of carbohydrates. The development of the gelatin made it possible to obtain a differentiated product, contributing to diversification in the food sector, providing a viable alternative for maintaining consumer health and reducing costs compared to desserts already available on the market.
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Cocos , Gelatina , Lacticaseibacillus rhamnosus , Probióticos , Cocos/química , Cocos/microbiología , Gelatina/química , Antioxidantes/farmacología , Antioxidantes/química , FermentaciónRESUMEN
INTRODUCTION AND OBJECTIVES: There are different situations in which an extrahepatic bile duct replacement or substitute is needed, such as initial and localized stages of bile duct cancer, agenesis, stenosis, or bile duct disruption. MATERIALS AND METHODS: A prosthesis obtained by electrospinning composed of Poly (D,L-lactide-co-glycolide) (PGLA) - Polycaprolactone (PCL) - Gelatin (Gel) was developed, mechanical and biological tests were carried out to evaluate resistance to tension, biocompatibility, biodegradability, cytotoxicity, morphological analysis and cell culture. The obtained prosthesis was placed in the extrahepatic bile duct of 15 pigs with a 2-year follow-up. Liver function tests and cholangioscopy were evaluated during follow-up. RESULTS: Mechanical and biological evaluations indicate that this scaffold is biocompatible and biodegradable. The prosthesis implanted in the experimental model allowed cell adhesion, migration, and proliferation, maintaining bile duct permeability without altering liver function tests. Immunohistochemical analysis indicates the presence of biliary epithelium. CONCLUSIONS: A tubular scaffold composed of electrospun PGLA-PCL-Gel nanofibers was used for the first time to replace the extrahepatic bile duct in pigs. Mechanical and biological evaluations indicate that this scaffold is biocompatible and biodegradable, making it an excellent candidate for use in bile ducts and potentially in other tissue engineering applications.
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Implantes Absorbibles , Conductos Biliares Extrahepáticos , Gelatina , Poliésteres , Ingeniería de Tejidos , Andamios del Tejido , Animales , Conductos Biliares Extrahepáticos/cirugía , Ingeniería de Tejidos/métodos , Porcinos , Ensayo de Materiales , Copolímero de Ácido Poliláctico-Ácido Poliglicólico/química , Proliferación Celular , Diseño de Prótesis , Materiales Biocompatibles , Movimiento Celular , Adhesión Celular , Factores de Tiempo , Pruebas de Función Hepática , NanofibrasRESUMEN
Bone tissue regeneration strategies have incorporated the use of natural polymers, such as hydroxyapatite (nHA), chitosan (CH), gelatin (GEL), or alginate (ALG). Additionally, platelet concentrates, such as platelet-rich fibrin (PRF) have been suggested to improve scaffold biocompatibility. This study aimed to develop scaffolds composed of nHA, GEL, and CH, with or without ALG and lyophilized PRF, to evaluate the scaffold's properties, growth factor release, and dental pulp stem cells (DPSC), and osteoblast (OB) derived from DPSC viability. Four scaffold variations were synthesized and lyophilized. Then, degradation, swelling profiles, and morphological analysis were performed. Furthermore, PDGF-BB and FGF-B growth factors release were quantified by ELISA, and cytotoxicity and cell viability were evaluated. The swelling and degradation profiles were similar in all scaffolds, with pore sizes ranging between 100 and 250 µm. FGF-B and PDGF-BB release was evidenced after 24 h of scaffold immersion in cell culture medium. DPSC and OB-DPSC viability was notably increased in PRF-supplemented scaffolds. The nHA-CH-GEL-PRF scaffold demonstrated optimal physical-biological characteristics for stimulating DPSC and OB-DPSC cell viability. These results suggest lyophilized PRF improves scaffold biocompatibility for bone tissue regeneration purposes.
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Alginatos , Supervivencia Celular , Quitosano , Pulpa Dental , Durapatita , Gelatina , Osteoblastos , Fibrina Rica en Plaquetas , Células Madre , Andamios del Tejido , Humanos , Pulpa Dental/citología , Quitosano/química , Quitosano/farmacología , Gelatina/química , Fibrina Rica en Plaquetas/química , Fibrina Rica en Plaquetas/metabolismo , Andamios del Tejido/química , Células Madre/efectos de los fármacos , Células Madre/citología , Células Madre/metabolismo , Supervivencia Celular/efectos de los fármacos , Durapatita/química , Durapatita/farmacología , Alginatos/química , Alginatos/farmacología , Osteoblastos/efectos de los fármacos , Osteoblastos/citología , Adhesión Celular/efectos de los fármacos , Ingeniería de Tejidos/métodos , Células CultivadasRESUMEN
Tumor-on-chips (ToCs) are useful platforms for studying the physiology of tumors and evaluating the efficacy and toxicity of anti-cancer drugs. However, the design and fabrication of a ToC system is not a trivial venture. We introduce a user-friendly, flexible, 3D-printed microfluidic device that can be used to culture cancer cells or cancer-derived spheroids embedded in hydrogels under well-controlled environments. The system consists of two lateral flow compartments (left and right sides), each with two inlets and two outlets to deliver cell culture media as continuous liquid streams. The central compartment was designed to host a hydrogel in which cells and microtissues can be confined and cultured. We performed tracer experiments with colored inks and 40 kDa fluorescein isothiocyanate dextran to characterize the transport/mixing performances of the system. We also cultured homotypic (MCF7) and heterotypic (MCF7-BJ) spheroids embedded in gelatin methacryloyl hydrogels to illustrate the use of this microfluidic device in sustaining long-term micro-tissue culture experiments. We further demonstrated the use of this platform in anticancer drug testing by continuous perfusion of doxorubicin, a commonly used anti-cancer drug for breast cancer. In these experiments, we evaluated drug transport, viability, glucose consumption, cell death (apoptosis), and cytotoxicity. In summary, we introduce a robust and friendly ToC system capable of recapitulating relevant aspects of the tumor microenvironment for the study of cancer physiology, anti-cancer drug transport, efficacy, and safety. We anticipate that this flexible 3D-printed microfluidic device may facilitate cancer research and the development and screening of strategies for personalized medicine.
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Antineoplásicos , Neoplasias de la Mama , Impresión Tridimensional , Esferoides Celulares , Humanos , Esferoides Celulares/efectos de los fármacos , Esferoides Celulares/patología , Esferoides Celulares/metabolismo , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/patología , Neoplasias de la Mama/metabolismo , Antineoplásicos/farmacología , Antineoplásicos/química , Femenino , Células MCF-7 , Hidrogeles/química , Dispositivos Laboratorio en un Chip , Línea Celular Tumoral , Ensayos de Selección de Medicamentos Antitumorales , Dextranos/química , Gelatina/química , Doxorrubicina/farmacología , Doxorrubicina/química , Supervivencia Celular/efectos de los fármacos , MetacrilatosRESUMEN
The difficulty in swallowing is a frequent problem when oral solid dosage forms (conventional tablets or capsules) are administered to paediatric population or patients with dysphagia. An interesting alternative to overcome these problems are non-conventional formulations like chewable gels, commonly known as 'gummies'. Therefore, this work addresses the design, development and characterization of gummies using gelatine and pectin, for the vehiculization of the antiarrhythmic amiodarone (AMIO). Applying a Design of Experiments (DoE) approach, four gelatine (GG1-GG4) and eight pectin formulations (PG1-PG8) were developed. Considering the obtained results for responses during DoE evaluation (i.e., volume, syneresis, hardness, and gumminess), GG3 and PG8 were selected for complete characterization. Water activity, pH, drug content, texture parameters (adhesiveness, springiness, cohesiveness, and fracturability), disintegration time, in vitro dissolution, and microbiological features were evaluated. The obtained results were within the expected values for this type of formulation. The dissolution profiles showed a 94 % - 99 % of the AMIO content released for GG3 and PG8, respectively, so they could be considered suitable as immediate release dosage forms. In conclusion, the chewable gels were successfully developed and characterised, suggesting a potential means to accomplish a final prototype for the improvement of congenital cardiopathies treatment.
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Amiodarona , Antiarrítmicos , Geles , Cardiopatías Congénitas , Pectinas , Amiodarona/administración & dosificación , Amiodarona/química , Humanos , Pectinas/química , Antiarrítmicos/administración & dosificación , Antiarrítmicos/química , Cardiopatías Congénitas/tratamiento farmacológico , Gelatina/química , Animales , Niño , Administración Oral , Liberación de Fármacos , Composición de Medicamentos/métodos , Solubilidad , Química Farmacéutica/métodosRESUMEN
This study aimed to evaluate the osteogenic potential of hydroxyapatite (HA), Alginate (Alg), and Gelatine (Gel) composite in a critical-size defect model in rats. Twenty-four male rats were divided into three groups: a negative control with no treatment (Control group), a positive control treated with deproteinized bovine bone mineral (DBBM group), and the experimental group treated with the new HA-Alg-Gel composite (HA-Alg-Gel group). A critical size defect (8.5mm) was made in the rat's calvaria, and the bone formation was evaluated by in vivo microcomputed tomography analysis (µCT) after 1, 15, 45, and 90 days. After 90 days, the animals were euthanized and histological and histomorphometric analyses were performed. A higher proportion of mineralized tissue/biomaterial was observed in the DBBM group when compared to the HA-Alg-Gel and Control groups in the µCT analysis during all analysis periods. However, no differences were observed in the mineralized tissue/biomaterial proportion observed on day 1 (immediate postoperative) in comparison to later periods of analysis in all groups. In the histomorphometric analysis, the HA-Alg-Gel and Control groups showed higher bone formation than the DBBM group. Moreover, in histological analysis, five samples of the HA-Alg-Gal group exhibited formed bone spicules adjacent to the graft granules against only two of eight samples in the DBBM group. Both graft materials ensured the maintenance of defect bone thickness, while a tissue thickness reduction was observed in the control group. In conclusion, this study demonstrated the osteoconductive potential of HA-Alg-Gel bone graft by supporting new bone formation around its particles.
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Alginatos , Regeneración Ósea , Durapatita , Gelatina , Cráneo , Microtomografía por Rayos X , Animales , Regeneración Ósea/efectos de los fármacos , Durapatita/farmacología , Cráneo/cirugía , Cráneo/diagnóstico por imagen , Ratas , Masculino , Materiales Biocompatibles , Ácido Glucurónico , Ratas Wistar , Ácidos Hexurónicos , Osteogénesis/efectos de los fármacos , Sustitutos de HuesosRESUMEN
The necessity of animal-free performance tests for novel ophthalmic formulation screening is challenging. For this, we developed and validated a new device to simulate the dynamics and physical-chemical barriers of the eye for in vitro performance tests of topic ophthalmic formulations. The OphthalMimic is a 3D-printed device with an artificial lacrimal flow, a cul-de-sac area, a support base, and a simulated cornea comprised of a polymeric membrane containing poly-vinyl alcohol 10 % (w/v), gelatin 2.5 % (w/v), and different proportions of mucin and poloxamer, i.e., 1:1 (M1), 1:2 (M2), and 2:1 (M3) w/v, respectively. The support base is designed to move between 0° and 50° to replicate the movement of an eyelid. We challenged the model by testing the residence performance of poloxamer®407 16 % and poloxamer®407 16 % + chitosan 1 % (PLX16CS10) gels containing fluconazole. The test was conducted with a simulated tear flow of 1.0 mL.min-1 for 5 min. The OphthalMimic successfully distinguished PLX16 and PLX16C10 formulations based on their fluconazole drainage (M1: 65 ± 14 % and 27 ± 10 %; M2: 58 ± 6 % and 38 ± 9 %; M3: 56 ± 5 % and 38 ± 18 %). In conclusion, the OphthalMimic is a promising tool for comparing the animal-free performance of ophthalmic formulations.
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Soluciones Oftálmicas , Poloxámero , Poloxámero/química , Soluciones Oftálmicas/química , Administración Oftálmica , Fluconazol/administración & dosificación , Impresión Tridimensional , Córnea/efectos de los fármacos , Córnea/metabolismo , Animales , Quitosano/química , Alternativas a las Pruebas en Animales/métodos , Lágrimas/química , Humanos , Gelatina/químicaRESUMEN
Mannheimiahaemolytica is an opportunistic agent of the respiratory tract of bovines, a member of the Pasteurellaceae family, and the causal agent of fibrinous pleuropneumonia. This bacterium possesses different virulence factors, allowing it to colonize and infect its host. The present work describes the isolation and characterization of a serine protease secreted by M. haemolytica serotype 1. This protease was isolated from M. haemolytica cultured media by precipitation with 50 % methanol and ion exchange chromatography on DEAE-cellulose. It is a 70-kDa protease able to degrade sheep and bovine fibrinogen or porcine gelatin but not bovine IgG, hemoglobin, or casein. Mass spectrometric analysis indicates its identity with protease IV of M. haemolytica. The proteolytic activity was active between pH 5 and 9, with an optimal pH of 8. It was stable at 50 °C for 10 min but inactivated at 60 °C. The sera of bovines with chronic or acute pneumonia recognized this protease. Still, it showed no cross-reactivity with rabbit hyperimmune serum against the secreted metalloprotease from Actinobacilluspleuropneumoniae, another member of the Pasteurellaceae family. M. haemolytica secreted proteases could contribute to the pathogenesis of this bacterium through fibrinogen degradation, a characteristic of this fibrinous pleuropneumonia.
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Fibrinógeno , Mannheimia haemolytica , Serina Proteasas , Animales , Mannheimia haemolytica/enzimología , Ovinos , Bovinos , Fibrinógeno/metabolismo , Concentración de Iones de Hidrógeno , Serina Proteasas/metabolismo , Serina Proteasas/aislamiento & purificación , Temperatura , Proteolisis , Peso Molecular , Gelatina/metabolismo , Estabilidad de Enzimas , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/aislamiento & purificación , Espectrometría de Masas , Cromatografía por Intercambio Iónico , Porcinos , Factores de Virulencia/metabolismo , Factores de Virulencia/aislamiento & purificaciónRESUMEN
3D-printed hydrogel scaffolds biomimicking the extracellular matrix (ECM) are key in cartilage tissue engineering as they can enhance the chondrogenic differentiation of mesenchymal stem cells (MSCs) through the presence of active nanoparticles such as graphene oxide (GO). Here, biomimetic hydrogels were developed by cross-linking alginate, gelatin, and chondroitin sulfate biopolymers in the presence of GO as a bioactive filler, with excellent processability for developing bioactive 3D printed scaffolds and for the bioprinting process. A novel bioink based on our hydrogel with embedded human MSCs presented a cell survival rate near 100% after the 3D bioprinting process. The effects of processing and filler concentration on cell differentiation were further quantitatively evaluated. The nanocomposited hydrogels render high MSC proliferation and viability, exhibiting intrinsic chondroinductive capacity without any exogenous factor when used to print scaffolds or bioprint constructs. The bioactivity depended on the GO concentration, with the best performance at 0.1 mg mL-1. These results were explained by the rational combination of the three biopolymers, with GO nanoparticles having carboxylate and sulfate groups in their structures, therefore, biomimicking the highly negatively charged ECM of cartilage. The bioactivity of this biomaterial and its good processability for 3D printing scaffolds and 3D bioprinting techniques open up a new approach to developing novel biomimetic materials for cartilage repair.
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Alginatos , Bioimpresión , Diferenciación Celular , Condrogénesis , Sulfatos de Condroitina , Gelatina , Hidrogeles , Células Madre Mesenquimatosas , Nanocompuestos , Impresión Tridimensional , Andamios del Tejido , Humanos , Células Madre Mesenquimatosas/efectos de los fármacos , Células Madre Mesenquimatosas/citología , Sulfatos de Condroitina/química , Sulfatos de Condroitina/farmacología , Alginatos/química , Alginatos/farmacología , Gelatina/química , Bioimpresión/métodos , Diferenciación Celular/efectos de los fármacos , Condrogénesis/efectos de los fármacos , Nanocompuestos/química , Andamios del Tejido/química , Hidrogeles/química , Hidrogeles/farmacología , Ingeniería de Tejidos/métodos , Materiales Biomiméticos/química , Materiales Biomiméticos/farmacología , Grafito/química , Grafito/farmacología , Proliferación Celular/efectos de los fármacos , Células CultivadasRESUMEN
Biomaterial scaffolds play a pivotal role in the advancement of cultured meat technology, facilitating essential processes like cell attachment, growth, specialization, and alignment. Currently, there exists limited knowledge concerning the creation of consumable scaffolds tailored for cultured meat applications. This investigation aimed to produce edible scaffolds featuring both smooth and patterned surfaces, utilizing biomaterials such as salmon gelatin, alginate, agarose and glycerol, pertinent to cultured meat and adhering to food safety protocols. The primary objective of this research was to uncover variations in transcriptomes profiles between flat and microstructured edible scaffolds fabricated from marine-derived biopolymers, leveraging high-throughput sequencing techniques. Expression analysis revealed noteworthy disparities in transcriptome profiles when comparing the flat and microstructured scaffold configurations against a control condition. Employing gene functional enrichment analysis for the microstructured versus flat scaffold conditions yielded substantial enrichment ratios, highlighting pertinent gene modules linked to the development of skeletal muscle. Notable functional aspects included filament sliding, muscle contraction, and the organization of sarcomeres. By shedding light on these intricate processes, this study offers insights into the fundamental mechanisms underpinning the generation of muscle-specific cultured meat.
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Diferenciación Celular , Carne in Vitro , Andamios del Tejido , Transcriptoma , Animales , Alginatos/química , Materiales Biocompatibles/química , Biopolímeros , Gelatina/química , Perfilación de la Expresión Génica , Células Musculares/metabolismo , Desarrollo de Músculos/genética , Salmón , Sefarosa/química , Andamios del Tejido/químicaRESUMEN
Among non-communicable diseases, cardiovascular diseases are the most prevalent, accounting for approximately 17 million deaths per year. Despite conventional treatment, cardiac tissue engineering emerges as a potential alternative for the advancement and treatment of these patients, using biomaterials to replace or repair cardiac tissues. Among these materials, gelatin in its methacrylated form (GelMA) is a biodegradable and biocompatible polymer with adjustable biophysical properties. Furthermore, gelatin has the ability to replace and perform collagen-like functions for cell development in vitro. The interest in using GelMA hydrogels combined with nanomaterials is increasingly growing to promote the responsiveness to external stimuli and improve certain properties of these hydrogels by exploring the incorporation of nanomaterials into these hydrogels to serve as electrical signaling conductive elements. This review highlights the applications of electrically conductive nanomaterials associated with GelMA hydrogels for the development of structures for cardiac tissue engineering, by focusing on studies that report the combination of GelMA with nanomaterials, such as gold and carbon derivatives (carbon nanotubes and graphene), in addition to the possibility of applying these materials in 3D tissue engineering, developing new possibilities for cardiac studies.
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Gelatina , Nanotubos de Carbono , Humanos , Gelatina/química , Andamios del Tejido/química , Nanotubos de Carbono/química , Hidrogeles/química , Materiales Biocompatibles/química , Ingeniería de TejidosRESUMEN
The aim of this study was to evaluate the protective effect of the encapsulation of Limosilactobacillus reuteri DSPV002C in macrocapsules made from industrial materials during production, storage and under simulated gastrointestinal conditions in vitro and in vivo. The production of macrocapsules involved the evaluation of different wall materials (matrix), namely, gelatin and pregelatinized starch, different inoculums, matrix ratios, and diverse cryoprotectants (whey permeate and maltodextrin). The different macrocapsules were arranged in molds of similar size to pig pelleted food and lyophilized. Then, the viability of the macrocapsules was assessed over time during storage at different temperatures (freezing, refrigeration and room temperature) and atmospheres (vacuum and non-vaccum). The macrocapsules with 10% w/v gelatin+5% w/v pregelatinized starch, permeated (10%, w/v), with a 9:1 inoculum:matrix ratio (GS7.5P9), stored under freezing conditions and vacuum, exhibited the highest viability of L. reuteri DSPV002C (9.3 log CFU/cap until 210 d). Under simulated gastrointestinal conditions, the encapsulated inoculum showed less viability loss (0.58±0.09 log CFU/ml, 26.53%), compared to the free culture (1.56±0.16 log CFU/ml, 2.85%). Finally, by administering GS7.5P9 to pigs, the tolerance of the bacteria to the gastrointestinal environment was verified, with viable counts equal to or greater than 3.72 log CFU/g of fecal matter throughout the trial. In this study, a high-density carrier probiotic macrocapsule of L. reuteri DSPV002C was obtained, which displayed a long shelf life, a suitable shape to be included in pig feed and an adequate survival of viable cells at the site of action.
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Limosilactobacillus reuteri , Probióticos , Animales , Porcinos , Gelatina , Suplementos Dietéticos , AlmidónRESUMEN
Hydrogel filaments were manufactured using wet spinning technique, incorporating variations in the concentrations of sodium alginate, gelatin, and calcium chloride (crosslinking agent). The combination of biopolymer concentrations was determined using design of experiments (DoE) approach. The resulting filaments were produced from the developed hydrogels. Tensile and vertical strength analyses of the filaments were conducted using an electromechanical extensor. Atomic force microscopy was employed to evaluate the roughness, viscoelasticity, retraction, and deflection of the hydrogels. By employing DoE, a total of seventeen different combinations of biopolymers and crosslinkers were generated to construct the hydrogels. The filaments exhibited variations in electromechanical traction (measured in kPa) and produced distinct stress peaks. Furthermore, diverse roughness values were observed among the tested materials, with the combinations featuring higher concentrations of sodium alginate displaying the highest Young's modulus. This study demonstrates that manipulating the concentrations of biopolymers and crosslinking agents can modulate the micro and nanomechanical properties of biopolymeric filaments.
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Gelatina , Hidrogeles , Biopolímeros , Módulo de Elasticidad , AlginatosRESUMEN
In the present study, a film based on the gelatin skin of tilapia (Oreochromis niloticus) was developed, using surfactants and adding plant extract of pitomba seed (Talisia esculenta). The aim was to investigate the mechanical and barrier properties of the cover, as well as its effectiveness in conserving papayas against diseases caused by fungi. The film presented tensile strength of 38.78 MPa, elongation of 120.49%, and water vapor permeability of 5.90 g.mm.h-1.m2.kPa-1 when equally composed of SDS and Tween 80, in a percentage of 40% in relation to the total mass of the film. The films lasted 12 d in an environment with a relative humidity of 75% (25 ºC), longer than the shelf life of papaya (limited to 8 d). With applying the film with the extract, the incidence of diseases such as anthracnose, fusariosis, and stem rot caused by these microorganisms in papaya was reduced.