RESUMEN
Chitosan chemical functionalization is a powerful tool to provide novel materials for additive manufacturing strategies. The main aim of this study was the employment of computer-aided wet spinning (CAWS) for the first time to design and fabricate carboxymethyl chitosan (CMCS) scaffolds. For this purpose, the synthesis of a chitosan derivative with a high degree of O-substitution (1.07) and water soluble in a large pH range allowed the fabrication of scaffolds with a 3D interconnected porous structure. In particular, the developed scaffolds were composed of CMCS fibers with a small diameter (< 60 µm) and a hollow structure due to a fast non solvent-induced coagulation. Zn2+ ionotropic crosslinking endowed the CMCS scaffolds with stability in aqueous solutions, pH-sensitive water uptake capability, and antimicrobial activity against Escherichia coli and Staphylococcus aureus. In addition, post-printing functionalization through collagen grafting resulted in a decreased stiffness (1.6 ± 0.3 kPa) and a higher elongation at break (101 ± 9 %) of CMCS scaffolds, as well as in their improved ability to support in vitro fibroblast viability and wound healing process. The obtained results encourage therefore further investigation of the developed scaffolds as antimicrobial wound dressing hydrogels for skin regeneration.
Asunto(s)
Antibacterianos , Vendajes , Quitosano , Escherichia coli , Staphylococcus aureus , Andamios del Tejido , Cicatrización de Heridas , Quitosano/química , Quitosano/análogos & derivados , Quitosano/farmacología , Staphylococcus aureus/efectos de los fármacos , Escherichia coli/efectos de los fármacos , Cicatrización de Heridas/efectos de los fármacos , Andamios del Tejido/química , Antibacterianos/farmacología , Antibacterianos/química , Animales , Ratones , Fibroblastos/efectos de los fármacos , Porosidad , Supervivencia Celular/efectos de los fármacos , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Materiales Biocompatibles/síntesis química , Reactivos de Enlaces Cruzados/química , HumanosRESUMEN
Large bone defects are a significant health problem today with various origins, including extensive trauma, tumours, or congenital musculoskeletal disorders. Tissue engineering, and in particular bone tissue engineering, aims to respond to this demand. As such, we propose a specific model based on Elastin-Like Recombinamers-based click-chemistry hydrogels given their high biocompatibility and their potent on bone regeneration effect conferred by different bioactive sequences. In this work we demonstrate, using biochemistry, histology, histomorphometry and imaging techniques, the biocompatibility of our matrix and its potent effect on bone regeneration in a model of bone parietal lesion in female New Zealand rabbits.
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Materiales Biocompatibles , Regeneración Ósea , Elastina , Hidrogeles , Ingeniería de Tejidos , Animales , Femenino , Conejos , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Regeneración Ósea/efectos de los fármacos , Química Clic/métodos , Elastina/química , Hidrogeles/química , Hidrogeles/farmacología , Ingeniería de Tejidos/métodos , Andamios del Tejido/químicaRESUMEN
BACKGROUND: Tissue engineering seeks to improve, maintain, or replace the biological functions of damaged organs or tissues with biological substitutes such as the development of scaffolds. In the case of bone tissue, they must have excellent mechanical properties like native bone. OBJECTIVE: In this work, three geometric models were designed for scaffolds with different structure lattices and porosity that could be biomechanically suitable and support cell growth for trabecular bone replacement applications in tissue engineering and regenerative medicine to the proximal femur area. METHODS: Geometries were designed using computer-aided design (CAD) software and evaluated using finite element analysis in compression tests. Three loads were considered according to the daily activity: 1177 N for slow walking, 2060 N for fast walking, and 245.25 N for a person in a bipedal position. All these loads for an adult weight of 75 kg. For each of them, three biomaterials were assigned: two polymers (poly-glycolic acid (PGA) and poly-lactic acid (PLA)) and one mineral (hydroxyapatite (HA)). 54 tests were performed: 27 for each of the tests. RESULTS: The results showed Young's modulus (E) between 1 and 4 GPa. CONCLUSION: If the resultant E is in the range of 0.1 to 5 GPa, the biomaterial is considered an appropriate alternative for the trabecular bone which is the main component of the proximal bone. However, for the models applied in this study, the best option is the poly-lactic acid which will allow absorbing the acting loads.
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Diseño Asistido por Computadora , Análisis de Elementos Finitos , Ingeniería de Tejidos , Andamios del Tejido , Andamios del Tejido/química , Humanos , Ingeniería de Tejidos/métodos , Durapatita/química , Módulo de Elasticidad , Bioimpresión/métodos , Poliésteres/química , Porosidad , Simulación por Computador , Materiales Biocompatibles/química , Sustitutos de Huesos/química , Ácido Poliglicólico/química , Impresión Tridimensional , Ensayo de Materiales , HuesosRESUMEN
The skin is a tissue constantly exposed to the risk of damage, such as cuts, burns, and genetic disorders. The standard treatment is autograft, but it can cause pain to the patient being extremely complex in patients suffering from burns on large body surfaces. Considering that there is a need to develop technologies for the repair of skin tissue like 3D bioprinting. Skin is a tissue that is approximately 1/16 of the total body weight and has three main layers: epidermis, dermis, and hypodermis. Therefore, there are several studies using cells, biomaterials, and bioprinting for skin regeneration. Here, we provide an overview of the structure and function of the epidermis, dermis, and hypodermis, and showed in the recent research in skin regeneration, the main cells used, biomaterials studied that provide initial support for these cells, allowing the growth and formation of the neotissue and general characteristics, advantages and disadvantages of each methodology and the landmarks in recent research in the 3D skin bioprinting.
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Materiales Biocompatibles , Bioimpresión , Impresión Tridimensional , Medicina Regenerativa , Piel , Ingeniería de Tejidos , Humanos , Medicina Regenerativa/métodos , Materiales Biocompatibles/química , Ingeniería de Tejidos/métodos , Animales , Piel/citología , Andamios del Tejido/química , RegeneraciónRESUMEN
Materials with a soft tissue regenerative capacity can be produced using biopolymer scaffolds and nanomaterials, which allow injured tissue to recover without any side effects or limitations. Four formulations were prepared using polyvinyl alcohol (PVA) and chitosan (CS), with silicon dioxide nanoparticles (NPs-SiO2) incorporated using the freeze-drying method at a temperature of -50 °C. TGA and DSC showed no change in thermal degradation, with glass transition temperatures around 74 °C and 77 °C. The interactions between the hydroxyl groups of PVA and CS remained stable. Scanning electron microscopy (SEM) indicated that the incorporation of NPs-SiO2 complemented the freeze-drying process, enabling the dispersion of the components on the polymeric matrix and obtaining structures with a small pore size (between 30 and 60 µm) and large pores (between 100 and 160 µm). The antimicrobial capacity analysis of Gram-positive and Gram-negative bacteria revealed that the scaffolds inhibited around 99% of K. pneumoniae, E. cloacae, and S. aureus ATCC 55804. The subdermal implantation analysis demonstrated tissue growth and proliferation, with good biocompatibility, promoting the healing process for tissue restoration through the simultaneous degradation and formation of type I collagen fibers. All the results presented expand the boundaries in tissue engineering and regenerative medicine by highlighting the crucial role of nanoparticles in optimizing scaffold properties.
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Quitosano , Liofilización , Nanopartículas , Alcohol Polivinílico , Dióxido de Silicio , Ingeniería de Tejidos , Andamios del Tejido , Quitosano/química , Alcohol Polivinílico/química , Dióxido de Silicio/química , Andamios del Tejido/química , Nanopartículas/química , Ingeniería de Tejidos/métodos , Animales , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Antibacterianos/farmacología , Antibacterianos/química , Medicina Regenerativa/métodos , Regeneración/efectos de los fármacosRESUMEN
Pancreatic bioengineering is a potential therapeutic alternative for type 1 diabetes (T1D) in which the pancreas is decellularized, generating an acellular extracellular matrix (ECM) scaffold, which may be reconstituted by recellularization with several cell types to generate a bioartificial pancreas. No consensus for an ideal pancreatic decellularization protocol exists. Therefore, we aimed to determine the best-suited detergent by comparing sodium dodecyl sulfate (SDS), sodium deoxycholate (SDC), and Triton X-100 at different concentrations. Murine (n=12) and human pancreatic tissue from adult brain-dead donors (n=06) was harvested in accordance with Institutional Ethical Committee of the University of São Paulo Medical School (CEP-FMUSP) and decellularized under different detergent conditions. DNA content, histological analysis, and transmission and scanning electron microscopy were assessed. The most adequate condition for pancreatic decellularization was found to be 4% SDC, displaying: a) effective cell removal; b) maintenance of extracellular matrix architecture; c) proteoglycans, glycosaminoglycans (GAGs), and collagen fibers preservation. This protocol was extrapolated and successfully applied to human pancreas decellularization. The acellular ECM scaffold generated was recelullarized using human pancreatic islets primary clusters. 3D clusters were generated using 0.5×104 cells and then placed on top of acellular pancreatic slices (25 and 50 µm thickness). These clusters tended to connect to the acellular matrix, with visible cells located in the periphery of the clusters interacting with the ECM network of the bioscaffold slices and continued to produce insulin. This study provided evidence on how to improve and accelerate the pancreas decellularization process, while maintaining its architecture and extracellular structure, aiming at pancreatic bioengineering.
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Ácido Desoxicólico , Detergentes , Páncreas , Dodecil Sulfato de Sodio , Ingeniería de Tejidos , Andamios del Tejido , Animales , Detergentes/química , Detergentes/farmacología , Humanos , Páncreas/citología , Ratones , Dodecil Sulfato de Sodio/farmacología , Ácido Desoxicólico/farmacología , Ácido Desoxicólico/química , Andamios del Tejido/química , Ingeniería de Tejidos/métodos , Octoxinol/química , Matriz Extracelular , Diabetes Mellitus Tipo 1 , Microscopía Electrónica de Rastreo , Matriz Extracelular Descelularizada/químicaRESUMEN
Due to bioactive properties, introducing spongin-like collagen (SPG) into the biosilica (BS) extracted from marine sponges would present an enhanced biological material for improving osteoporotic fracture healing by increasing bone formation rate. Our aim was to characterize the morphology of the BS/SPG scaffolds by scanning electron microscopy (SEM), the chemical bonds of the material by Fourier transform infrared spectroscopy (FTIR), and evaluating the orthotopic in vivo response of BS/SPG scaffolds in tibial defects of osteoporotic fractures in rats (histology, histomorphometry, and immunohistochemistry) in two experimental periods (15 and 30 days). SEM showed that scaffolds were porous, showing the spicules of BS and fibrous aspect of SPG. FTIR showed characteristic peaks of BS and SPG. For the in vivo studies, after 30 days, BS and BS/SPG showed a higher amount of newly formed bone compared to the first experimental period, observed both in the periphery and in the central region of the bone defect. For histomorphometry, BS/SPG presented higher %BV/TV compared to the other experimental groups. After 15 days, BS presented higher volumes of collagen type I. After 30 days, all groups demonstrated higher volumes of collagen type III compared to volumes at 15 days. After 30 days, BS/SPG presented higher immunostaining of osteoprotegerin compared to the other experimental groups at the same experimental period. The results showed that BS and BS/SPG scaffolds were able to improve bone healing. Future research should focus on the effects of BS/SPG on longer periods in vivo studies.
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Colágeno , Poríferos , Andamios del Tejido , Animales , Ratas , Andamios del Tejido/química , Poríferos/química , Colágeno/metabolismo , Femenino , Dióxido de Silicio/química , Osteoporosis/patología , Ratas Wistar , Fracturas Osteoporóticas , Microscopía Electrónica de Rastreo , Osteogénesis/efectos de los fármacos , Espectroscopía Infrarroja por Transformada de Fourier , TibiaRESUMEN
Scaffolds are 3D biomaterials that provide an environment for cell regeneration. In the context of bone remodeling, poly(e-caprolactone) (PCL) combined with graphene has been developed as the scaffold. It is imperative for scaffolds to possess antibacterial properties in order to properly reduce the risk of potential infections.Therefore, this study aims to analyze the antibacterial characteristics of PCL/graphene scaffolds against Staphylococcus aureus (S. aureus) and Porphyromonas gingivalis (P. gingivalis) in vitro. In this study, five different groups were used, including PCL (K-), Amoxicillin (K+), PCL/Graphene 0.5 wt%, PCL/graphene 1 wt% and PCL/Graphene 1.5 wt%. All experiments were performed in triplicates and were repeated three times, and the diffusion method by Kirby-Bauer test was used. The disc was incubated with S. aureus and P. gingivalis for 24 hours and then the diameter of the inhibition zone was measured. The results showed that the PCL/graphene scaffolds exhibited dose-dependent antibacterial activity against S. aureus and P. gingivalis. The inhibition zone diameter (IZD) against S. aureus of PCL/graphene 1 wt% was 9.53 ± 0.74 mm, and increased to 11.93 ± 0.92 mm at a concentration of 1.5 wt% of graphene. The PCL/graphene scaffold with 1.5 wt% exhibited a greater inhibitory effect, with an IZD of 12.56 ± 0.06 mm against P. gingivalis, while the inhibitory activity of the 1 wt% variant was relatively lower at 10.46 ± 0.24 mm. The negative control, PCL, and PCL/graphene 0.5 wt% exhibited no antibacterial activity sequentially (p = 1). Scaffolds of poly(e-caprolactone)/graphene exhibited an antibacterial activity at 1, and 1.5 wt% on S. aureus and P. gingivalis. The antibacterial properties of this scaffold make it a promising candidate for regenerating bone tissue.
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Antibacterianos , Grafito , Poliésteres , Porphyromonas gingivalis , Staphylococcus aureus , Andamios del Tejido , Grafito/química , Grafito/farmacología , Porphyromonas gingivalis/efectos de los fármacos , Staphylococcus aureus/efectos de los fármacos , Andamios del Tejido/química , Antibacterianos/farmacología , Antibacterianos/química , Poliésteres/química , Poliésteres/farmacología , Regeneración Ósea/efectos de los fármacos , Materiales Biocompatibles/farmacología , Materiales Biocompatibles/química , Pruebas de Sensibilidad MicrobianaRESUMEN
The objective of the present study was to evaluate the carbon fiber obtained from textile PAN fiber, in its different forms, as a potential scaffolds synthetic bone. Thirty-four adult rats were used (Rattus norvegicus, albinus variation), two critical sized bone defects were made that were 5 mm in diameter. Twenty-four animals were randomly divided into four groups: control (C)-bone defect + blood clot, non-activated carbon fiber felt (NACFF)-bone defect + NACFF, activated carbon fiber felt (ACFF)-bone defect + ACFF, and silver activated carbon fiber felt (Ag-ACFF)-bone defect + Ag-ACFF, and was observed by 15 and 60 days for histomorphometric, three-dimensional computerized microtomography (microCT) and mineral apposition analysis. On histomorphometric and microCT analyses, NACFF were associated with higher proportion of neoformed bone and maintenance of bone structure. On fluorochrome bone label, there was no differences between the groups. NACFF has shown to be a promising synthetic material as a scaffold for bone regeneration.
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Regeneración Ósea , Fibra de Carbono , Carbono , Andamios del Tejido , Microtomografía por Rayos X , Animales , Ratas , Regeneración Ósea/efectos de los fármacos , Andamios del Tejido/química , Carbono/química , Fibra de Carbono/química , Masculino , Textiles , Brasil , Ensayo de Materiales , Ratas WistarRESUMEN
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
Bioactive and biodegradable scaffolds that mimic the natural extracellular matrix of bone serve as temporary structures to guide new bone tissue growth. In this study, 3D-printed scaffolds composed of poly (lactic acid) (PLA)-tricalcium phosphate (TCP) (90-10 wt.%) were modified with 1%, 5%, and 10 wt.% of ZnO to enhance bone tissue regeneration. A commercial chain extender named Joncryl was incorporated alongside ZnO to ensure the printability of the composites. Filaments were manufactured using a twin-screw extruder and subsequently used to print 3D scaffolds via fused filament fabrication (FFF). The scaffolds exhibited a homogeneous distribution of ZnO and TCP particles, a reproducible structure with 300 µm pores, and mechanical properties suitable for bone tissue engineering, with an elastic modulus around 100 MPa. The addition of ZnO resulted in enhanced surface roughness on the scaffolds, particularly for ZnO microparticles, achieving values up to 241 nm. This rougher topography was responsible for enhancing protein adsorption on the scaffolds, with an increase of up to 85% compared to the PLA-TCP matrix. Biological analyses demonstrated that the presence of ZnO promotes mesenchymal stem cell (MSC) proliferation and differentiation into osteoblasts. Alkaline phosphatase (ALP) activity, an important indicator of early osteogenic differentiation, increased up to 29%. The PLA-TCP composite containing 5% ZnO microparticles exhibited an optimized degradation rate and enhanced bioactivity, indicating its promising potential for bone repair applications.
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Materiales Biocompatibles , Regeneración Ósea , Fosfatos de Calcio , Diferenciación Celular , Proliferación Celular , Células Madre Mesenquimatosas , Osteoblastos , Poliésteres , Impresión Tridimensional , Ingeniería de Tejidos , Andamios del Tejido , Óxido de Zinc , Andamios del Tejido/química , Fosfatos de Calcio/química , Poliésteres/química , Regeneración Ósea/efectos de los fármacos , Ingeniería de Tejidos/métodos , Células Madre Mesenquimatosas/citología , Óxido de Zinc/química , Materiales Biocompatibles/química , Diferenciación Celular/efectos de los fármacos , Osteoblastos/citología , Osteogénesis/efectos de los fármacos , Ensayo de Materiales , Huesos , Regeneración Tisular Dirigida/métodos , Humanos , Animales , Fosfatasa Alcalina/metabolismo , Módulo de Elasticidad , Porosidad , Propiedades de SuperficieRESUMEN
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
Hybrid scaffolds that are based on PLA and PLA/PMMA with 75/25, 50/50, and 25/75 weight ratios and functionalized with 10 wt.% of bioglass nanoparticles (n-BG) were developed using an electrospinning technique with a chloroform/dimethylformamide mixture in a 9:1 ratio for bone tissue engineering applications. Neat PLA and PLA/PMMA hybrid scaffolds were developed successfully through a (CF/DMF) solvent system, obtaining a random fiber deposition that generated a porous structure with pore interconnectivity. However, with the solvent system used, it was not possible to generate fibers in the case of the neat PMMA sample. With the increase in the amount of PMMA in PLA/PMMA ratios, the fiber diameter of hybrid scaffolds decreases, and the defects (beads) in the fiber structure increase; these beads are associated with a nanoparticle agglomeration, that could be related to a low interaction between n-BG and the polymer matrix. The Young's modulus of PLA/PMMA/n-BG decreases by 34 and 80%, indicating more flexible behavior compared to neat PLA. The PLA/PMMA/n-BG scaffolds showed a bioactive property related to the presence of hydroxyapatite crystals in the fiber surface after 28 days of immersion in a Simulated Body Fluids solution (SBF). In addition, the hydrolytic degradation process of PLA/PMMA/n-BG, analyzed after 35 days of immersion in a phosphate-buffered saline solution (PBS), was less than that of the pure PLA. The in vitro analysis using an HBOF-1.19 cell line indicated that the PLA/PMMA/n-BG scaffold showed good cell viability and was able to promote cell proliferation after 7 days. On the other hand, the in vivo biocompatibility evaluated via a subdermal model in BALC male mice corroborated the good behavior of the scaffolds in avoiding the generation of a cytotoxic effect and being able to enhance the healing process, suggesting that the materials are suitable for potential applications in tissue engineering.
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Cerámica , Nanopartículas , Poliésteres , Polimetil Metacrilato , Ingeniería de Tejidos , Andamios del Tejido , Ingeniería de Tejidos/métodos , Poliésteres/química , Polimetil Metacrilato/química , Andamios del Tejido/química , Cerámica/química , Cerámica/farmacología , Nanopartículas/química , Animales , Ratones , Huesos/efectos de los fármacos , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Humanos , Línea CelularRESUMEN
Bone tissue engineering is a promising alternative to repair wounds caused by cellular or physical accidents that humans face daily. In this sense, the search for new graphene oxide (GO) nanofillers related to their degree of oxidation is born as an alternative bioactive component in forming new scaffolds. In the present study, three different GOs were synthesized with varying degrees of oxidation and studied chemically and tissue-wise. The oxidation degree was determined through infrared (FTIR), X-ray diffraction (XRD), X-ray photoelectron (XPS), and Raman spectroscopy (RS). The morphology of the samples was analyzed using scanning electron microscopy (SEM). The oxygen content was deeply described using the deconvolution of RS and XPS techniques. The latter represents the oxidation degree for each of the samples and the formation of new bonds promoted by the graphitization of the material. In the RS, two characteristic bands were observed according to the degree of oxidation and the degree of graphitization of the material represented in bands D and G with different relative intensities, suggesting that the samples have different crystallite sizes. This size was described using the Tuinstra-Koenig model, ranging between 18.7 and 25.1 nm. Finally, the bone neoformation observed in the cranial defects of critical size indicates that the F1 and F2 samples, besides being compatible and resorbable, acted as a bridge for bone healing through regeneration. This promoted healing by restoring bone and tissue structure without triggering a strong immune response.
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Regeneración Ósea , Grafito , Ingeniería de Tejidos , Andamios del Tejido , Grafito/química , Regeneración Ósea/efectos de los fármacos , Ingeniería de Tejidos/métodos , Animales , Andamios del Tejido/química , Nanoestructuras/química , Huesos/efectos de los fármacos , Espectrometría Raman , Oxidación-Reducción , Difracción de Rayos X , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Ratas , Espectroscopía Infrarroja por Transformada de FourierRESUMEN
This study focuses on designing and evaluating scaffolds with essential properties for bone regeneration, such as biocompatibility, macroporous geometry, mechanical strength, and magnetic responsiveness. The scaffolds are made using 3D printing with acrylic resin and iron oxides synthesized through solution combustion. Utilizing triply periodic minimal surfaces (TPMS) geometry and mask stereolithography (MSLA) printing, the scaffolds achieve precise geometrical features. The mechanical properties are enhanced through resin curing, and magnetite particles from synthesized nanoparticles and alluvial magnetite are added for magnetic properties. The scaffolds show a balance between stiffness, porosity, and magnetic responsiveness, with maximum compression strength between 4.8 and 9.2 MPa and Young's modulus between 58 and 174 MPa. Magnetic properties such as magnetic coercivity, remanence, and saturation are measured, with the best results from scaffolds containing synthetic iron oxides at 1% weight. The viscosity of the mixtures used for printing is between 350 and 380 mPas, and contact angles between 90° and 110° are achieved. Biocompatibility tests indicate the potential for clinical trials, though further research is needed to understand the impact of magnetic properties on cellular interactions and optimize scaffold design for specific applications. This integrated approach offers a promising avenue for the development of advanced materials capable of promoting enhanced bone regeneration.
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Regeneración Ósea , Impresión Tridimensional , Andamios del Tejido , Andamios del Tejido/química , Porosidad , Ingeniería de Tejidos/métodos , Humanos , Materiales Biocompatibles/química , Compuestos Férricos/química , Fenómenos Magnéticos , Animales , MagnetismoRESUMEN
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
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
MicroRNAs (miRNAs) are non-coding RNAs involved in the regulation of gene expression associated with cell differentiation, proliferation, adhesion, and important biological functions such as inflammation. miRNAs play roles associated with the pathogenesis of chronic degenerative disorders including cardiovascular diseases. Understanding the influence of miRNAs and their target genes can effectively streamline the identification of key biologically active pathways that are important in the development of vascular grafts through the tissue engineering of blood vessels. To determine miRNA expression levels and identify miRNA target genes and pathways with biological roles in scaffolds that have been repopulated with adipose-derived stem cells (ASCs) generated through tissue engineering for the construction of blood vessels. miRNA quantification assays were performed in triplicate to determine miRNA expression in a total of 20 samples: five controls (natural inferior vena cava), five scaffolds recellularized with ASCs and differentiated into the endothelium (luminal layer), five samples of complete scaffolds seeded with ASCs differentiated into the endothelium (luminal layer) and smooth muscle (extraluminal layer), and five samples of ASC without cell differentiation. Several differentially expressed miRNAs were identified and predicted to modulate target genes with roles in key pathways associated with angiogenesis, vascular system control, and endothelial and smooth muscle regulation, including migration, proliferation, and growth. These findings underscore the involvement of these pathways in the regulatory mechanisms that are essential for vascular scaffold production through tissue engineering. Our research contributes to the knowledge of miRNA-regulated mechanisms, which may impact the design of vascular substitutes, and provide valuable insights for enhancing clinical practice. The molecular pathways regulated by miRNAs in tissue engineering of blood vessels (TEBV) allowed us to elucidate the main phenomena involved in cellular differentiation to constitute a blood vessel, with the main pathways being essential for angiogenesis, cellular differentiation, and differentiation into vascular smooth muscle.
Asunto(s)
Diferenciación Celular , MicroARNs , Ingeniería de Tejidos , Andamios del Tejido , MicroARNs/genética , MicroARNs/metabolismo , Ingeniería de Tejidos/métodos , Humanos , Andamios del Tejido/química , Diferenciación Celular/genética , Tejido Adiposo/metabolismo , Tejido Adiposo/citología , Vasos Sanguíneos/metabolismo , Vasos Sanguíneos/crecimiento & desarrollo , Regulación de la Expresión Génica , Neovascularización Fisiológica/genética , Células Madre/metabolismo , Células Madre/citología , Proliferación Celular/genética , Transducción de SeñalRESUMEN
Electrospun nanocomposite scaffolds with improved bioactive and biological properties were fabricated from a blend of polycaprolactone (PCL) and starch, and then combined with 5 wt% of calcium oxide (CaO) nanoparticles sourced from eggshells. SEM analyses showed scaffolds with fibrillar morphology and a three-dimensional structure. The hydrophilicity of scaffolds was improved with starch and CaO nanoparticles, which was evidenced by enhanced water absorption (3500 %) for 7 days. In addition, PCL/Starch/CaO scaffolds exhibited major degradation, with a mass loss of approximately 60 % compared to PCL/Starch and PCL/CaO. The PCL/Starch/CaO scaffolds decreased in crystallinity as intermolecular interactions between the nanoparticles retarded the mobility of the polymeric chains, leading to a significant increase in Young's modulus (ca. 60 %) and a decrease in tensile strength and elongation at break, compared to neat PCL. SEM-EDS, FT-IR, and XRD analyses indicated that PCL/Starch/CaO scaffolds presented a higher biomineralization capacity due to the ability to form hydroxyapatite (HA) in their surface after 28 days. The PCL/Starch/CaO scaffolds showed attractive biological performance, allowing cell adhesion and viability of M3T3-E1 preosteoblastic cells. In vivo analysis using a subdermal dorsal model in Wistar rats showed superior biocompatibility and improved resorption process compared to a pure PCL matrix. This biological analysis suggested that the PCL/Starch/CaO electrospun mats are suitable scaffolds for guiding the regeneration of bone tissue.
Asunto(s)
Huesos , Compuestos de Calcio , Nanopartículas , Óxidos , Poliésteres , Almidón , Ingeniería de Tejidos , Andamios del Tejido , Almidón/química , Poliésteres/química , Ingeniería de Tejidos/métodos , Andamios del Tejido/química , Animales , Nanopartículas/química , Óxidos/química , Compuestos de Calcio/química , Ratas , Ratones , Materiales Biocompatibles/química , Ratas Wistar , Línea Celular , Nanocompuestos/químicaRESUMEN
The objective of this study was to synthesize and characterize porous Cellulose Acetate (CA) scaffolds using the electrospinning technique and functionalize the surface of the scaffolds obtained through the dip-coating method with a Hydroxyapatite (HA) nanocomposite and varying concentrations of graphene oxide (GO) for application in tissue engineering regeneration techniques. The scaffolds were divided into four distinct groups based on their composition: 1) CA scaffolds; 2) CAHAC scaffolds; 3) CAHAGOC 1.0% scaffolds; 4) CAHAGOC 1.5% scaffolds. Scaffold analyses were conducted using X-ray Diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR), Raman Spectroscopy, Scanning Electron Microscopy with Energy Dispersive Spectroscopy (SEM/EDS), and in vitro cell viability assays (WST). For the biological test analysis, Variance (two-way) was used, followed by Tukey's post-test (α = 0.05). The XRD results revealed the predominant presence of CaP phases in the CAHAC, CAHAGOC 1.0%, and CAHAGOC 1.5% groups, emphasizing the presence of HA in the scaffolds. FTIR demonstrated characteristics of cellulose and PO4 bands in the groups containing HA, confirming the presence of CaP in the synthesized materials, as also indicated by XRD. Raman spectroscopy showed the presence of D and G bands, consistent with GO, confirming the successful incorporation of the HAGO nanocomposite into the scaffolds. The micrographs displayed overlapping electrospun fibers, forming the three-dimensional structure in the produced scaffolds. It was possible to observe hydroxyapatite crystals filling some of these pores, creating a suitable structure for cell adhesion, proliferation, and nutrition, as corroborated by the results of in vitro tests. All scaffolds exhibited high cell viability, with significant cell proliferation. Even after 48 h, there was a slight reduction in the number of cells, but a noteworthy increase in cell proliferation was evident in the CAHAGOC 1.5% group after 48 h (p < 0.05). In conclusion, it can be affirmed that the produced scaffolds demonstrated physical and biological characteristics and properties capable of promoting cell adhesion and proliferation. Therefore, they represent significant potential for application in tissue engineering, offering a new perspective regarding techniques and biomaterials applied in regenerative therapies.