RESUMO
Injectable bone substitute (IBS) materials are commonly used to fill irregular-shaped bone voids in non-load-bearing areas and can offer greater utility over those which are in prefabricated powder, granule, or block forms. This work investigates the impact of liquid-to-solid ratio (LSR) on the rheology and cytocompatibility of IBSs formulated from bioactive glass particles and ß-tricalcium phosphate (ß-TCP) in glycerol and poly(ethylene glycol) (PEG). IBS formulations of varying LSR were prepared and packed in 3 cc open-bore syringes and sterilized via gamma irradiation (10 kGy, 25 kGy). Gamma-irradiated formulations with high PEG content required the highest (73 N) mechanical force for injection from syringes. Oscillatory viscosity measurements revealed that the viscosity of samples was directly proportional to glycerol content. PEG and glycerol displayed competing effects on the washout resistance and cohesiveness of samples, which were based on total weight loss in media and Ca2+ ion release, respectively. Cell viability in 24-h extracts of 10 kGy gamma-sterilized and 25 kGy gamma-irradiated samples were 22.94% and 56.53%, respectively. The research highlights the complex interplay of IBS components on IBS rheology and, moreover, the cytotoxicity behaviors of beta-tricalcium phosphate-based injectable bone substitutes by in vitro experiments.
Assuntos
Substitutos Ósseos , Fosfatos de Cálcio , Sobrevivência Celular , Raios gama , Injeções , Teste de Materiais , Polietilenoglicóis , Reologia , Fosfatos de Cálcio/química , Substitutos Ósseos/química , Substitutos Ósseos/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Polietilenoglicóis/química , Animais , Camundongos , Viscosidade , Glicerol/química , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologiaRESUMO
Decellularized tissue hydrogels, especially that mimic the native tissue, have a high potential for tissue engineering, three-dimensional (3D) cell culture, bioprinting, and therapeutic agent encapsulation due to their excellent biocompatibility and ability to facilitate the growth of cells. It is important to note that the decellularization process significantly affects the structural integrity and properties of the extracellular matrix, which in turn shapes the characteristics of the resulting hydrogels at the macromolecular level. Therefore, our study aims to identify an effective chemical decellularization method for sheep lung tissue, using a mixing/agitation technique with a range of detergents, including commonly [Sodium dodecyl sulfate (SDS), Triton X-100, and 3-((3-cholamidopropyl) dimethylammonio)-1-propanesulfonate] (CHAPS), and rarely used (sodium cholate hydrate, NP-40, and 3-[N,N-Dimethyl(3-myristoylaminopropyl)ammonio]propanesulfonate) (ASB-14). After the effectiveness of the used detergents on decellularization was determined by histological and biochemical methods, lung derived decellularized extracellular matrix was converted into hydrogel. We investigated the interactions between lung cells and decellularized extracellular matrix using proliferation assay, scanning electron microscopy, and immunofluorescence microscopy methods on BEAS-2B cells in air-liquid interface. Notably, this study emphasizes the effectiveness of ASB-14 in the decellularization process, showcasing its crucial role in removing cellular components while preserving vital extracellular matrix biological macromolecules, including glycosaminoglycans, collagen, and elastin. The resulting hydrogels demonstrated favorable mechanical properties and are compatible with both cell-cell and cell-extracellular matrix interactions.
RESUMO
To search for a suitable meniscus repair material, acellular hybrid scaffolds consisting of in situ cross-linkable 3-D interpenetrating network structures were obtained by decellularization of the meniscus tissues followed by integration of the gel system. Decellularization efficiency was confirmed using a DNA quantification assay (82% decrease in DNA content) and histological stainings. In the second part of the study, the gelatin molecule was functionalized by adding methacrylic anhydride and the degree of functionalization was found to be 75% by (Proton-Nuclear Magnetic Resonance) 1H-NMR. Using this, a series of hybrid constructs named GelMA-Hybrid (G-Hybrid), GELMA/PEGDMA-Hybrid (PG-Hybrid), and GelMA/PEGDMA/HAMA-Hybrid (PGH-Hybrid) were prepared by cross-linking with UVA. Changes in the chemical structure were determined with Fourier Transform Infrared Spectrophotometer (FTIR). Water uptake capacities of cross-linked hybrid structures were measured in swelling studies, and it was found that hybrid scaffolds showed similar swelling properties compared to native counterparts. By compressive mechanical tests, enhanced mechanical properties were revealed in cross-linked scaffolds with PGH-Hybrid having the highest cross-link density. Protein denaturation and decomposition transition temperatures were improved by adding hydrogels to acellular scaffolds according to thermal gravimetric analyses (TGA). Cross-linked acellular scaffolds have exhibited a behavior close to native tissues with below 25% mass loss in phosphate buffer saline (PBS) and enzymatic solution. Cell viability was examined through Alamar Blue on the first day and cell viability in hybrid constructs was found to be above 80% while it was closer to the control group on the 7th day. It was concluded that the developed biomaterials could be used in meniscus tissue engineering with their tunable physicochemical and mechanical properties.
Assuntos
Menisco , Alicerces Teciduais , Alicerces Teciduais/química , Hidrogéis/química , Engenharia Tecidual , DNA , Gelatina/químicaRESUMO
Myocardial tissue engineering strategies such as fabrication of cardiac patches for tissue regeneration offer various solutions for the loss of function developed due to myocardial infarction. Here, we combined the hybrid structure (previously obtained and combined decellularized myocardium grafts with poly(glycerol-sebacate) polymer) with multiwalled carbon nanotubes (MWCNTs) to provide the essential characteristics for cardiac tissue regeneration. MWCNTs were doped in the cross-linked structure, and the conductivity and Young's modulus of the composite elastomer were found as 5 × 10-3 ± 1 × 10-3 S/m and 374 ± 75.8 kPa, respectively. The cell-material interaction was evaluated, and composite structures supported cell adhesion and showed no cytotoxic effect.
Assuntos
Nanotubos de Carbono , Nanotubos de Carbono/toxicidade , Nanotubos de Carbono/química , Miocárdio , Elastômeros/química , Engenharia Tecidual , Matriz ExtracelularRESUMO
Thrombogenicity, which is commonly encountered in artificial heart valves after replacement surgeries, causes valvular failure. Even life-long anticoagulant drug use may not be sufficient to prevent thrombogenicity. In this study, it was aimed to develop a heart valve construct with antithrombogenic properties and suitable mechanical strength by combining multiwalled carbon nanotubes within a decellularized bovine pericardium. In this context, the decellularization process was performed by using the combination of freeze-thawing and sodium dodecyl sulfate (SDS). Evaluation of decellularization efficiency was determined by histology (Hematoxylin and Eosin, DAPI and Masson's Trichrome) and biochemical (DNA, sGAG and collagen) analyses. After the decellularization process of the bovine pericardium, composite pericardial tissues were prepared by incorporating -COOH-modified multiwalled carbon nanotubes (MWCNTs). Characterization of MWCNT incorporation was performed by ATR-FTIR, TGA, and mechanical analysis, while SEM and AFM were used for morphological evaluations. Thrombogenicity assessments were studied by platelet adhesion test, Calcein-AM staining, kinetic blood clotting, hemolysis, and cytotoxicity analyses. As a result of this study, the composite pericardial material revealed improved mechanical and thermal stability and hemocompatibility in comparison to decellularized pericardium, without toxicity. Approximately 100% success is achieved in preventing platelet adhesion. In addition, kinetic blood-coagulation analysis demonstrated a low rate and slow coagulation kinetics, while the hemolysis index was below the permissible limit for biomaterials.
Assuntos
Próteses Valvulares Cardíacas , Nanocompostos , Nanotubos de Carbono , Animais , Bovinos , Valvas Cardíacas , HemóliseRESUMO
Introducing functional synthetic biomaterials to the literature became quite essential in biomedical technologies. For the growth of novel biomedical engineering approaches, progressive functional properties as well as the robustness of the manufacturing processes are essential. By using acid-induced epoxide ring-opening polymerizations through catalysts, a wide variety of biodegradable and functionalized biomaterials can be synthesized. Sebacic acid (SA) and poly(ethylene glycol) diglycidyl ether (PEGDGE) are amongst the FDA-approved biocompatible materials. In this study, we focused on the rapid synthesis of caffeine-catalyzed self-healable elastomer via a facile microwave-assisted synthesis route. The elastomer prepared can be used in various applications, including tactile sensors, wearable electronics, and soft robotics. SA and PEGDGE were catalyzed in the presence of caffeine under microwave irradiation followed by crosslinking in vacuo, yielding an elastomeric material. The chemical characterizations of the obtained elastomer were carried out. The resulting material is transparent, highly stretchable, and has capacitive and self-healing properties even at room temperature. The material developed can be easily applied for the aforementioned applications.
Assuntos
Elastômeros , Micro-Ondas , Materiais Biocompatíveis , Cafeína , Elastômeros/química , EtilenoglicolRESUMO
Supercritical carbon dioxide (scCO2) is being used as an alternative approach to the traditional methods for the decellularization of tissues. This chapter describes the use of scCO2 for the decellularization of optic nerve, myocardium, and cornea tissues. The main goal of this method is to burst the cells with high-pressure, remove them from the tissues and to maintain the extracellular matrix structure of the native tissues. For this purpose, several scCO2-assisted decellularization protocols were developed and optimized according to the requirements of these tissues. Efficiencies of the utilized decellularization protocols were determined via histological and morphological analysis. The decrease in the DNA content and the preserved glycosaminoglycan (GAG) amounts were also used as assessment parameters.
Assuntos
Dióxido de Carbono/química , Matriz Extracelular , Engenharia Tecidual/métodos , Alicerces Teciduais , Animais , Bovinos , Córnea/citologia , Glicosaminoglicanos/análise , Miocárdio/citologia , Nervo Óptico/citologiaRESUMO
The decellularization protocols applied on the corneal stromal constructs in the literature usually fail to provide a corneal matrix with sufficient mechanical and optical properties since they alter the extracellular matrix (ECM) microstructure. In this study, to overcome these limitations, a hybrid cornea stromal construct was engineered by combining gelatin methacrylate (GelMA) and decellularized ECM. Photo-cross-linking of impregnated cell laden GelMA in situ using different UV cross-linking energies (3200, 6210, and 6900 µJ/cm2) and impregnation times (up to 24 h) within a decellularized bovine cornea enhanced light transmission and restored lost mechanical features following the harsh decellularization protocol. The light transmittance value for optimized hybrid constructs (53.6%) was increased nearly 10 fold compared to that of decellularized cornea (5.84%). The compressive modulus was also restored up to 6 fold with calculated values of 5040 and 870 kPa for the hybrid and decellularized samples, respectively. These values were found to be quite close to that of native cornea (48.5%, 9790 kPa). ATR-FTIR analyses were carried out to confirm the final chemical structure. The degradation profiles showed similar decomposition behaviors to that of native cornea. In vitro culture studies showed a high level of cell viability and cell proliferation rate was found remarkable up to the 14th day of the culture period regardless of selected UV energy level. The methodology used in the preparation of the hybrid cornea stromal constructs in this study is a promising approach toward the development of successful corneal transplants.
Assuntos
Córnea , Hidrogéis , Animais , Bovinos , Matriz Extracelular , Gelatina , MetacrilatosRESUMO
Monodisperse silica microspheres with bimodal pore-size distribution were proposed as a high performance sorbent for DNA isolation in batch fashion under equilibrium conditions. The proposed sorbent including both macroporous and mesoporous compartments was synthesized 5.1 µm in-size, by a "staged shape templated hydrolysis and condensation method". Hydrophilic polymer based sorbents were also obtained in the form of monodisperse-macroporous microspheres ca 5.5 µm in size, with different functionalities, by a developed "multi-stage microsuspension copolymerization" technique. The batch DNA isolation performance of proposed material was comparatively investigated using polymer based sorbents with similar morphologies. Among all sorbents tried, the best DNA isolation performance was achieved with the monodisperse silica microspheres with bimodal pore size distribution. The collocation of interconnected mesoporous and macroporous compartments within the monodisperse silica microspheres provided a high surface area and reduced the intraparticular mass transfer resistance and made easier both the adsorption and desorption of DNA. Among the polymer based sorbents, higher DNA isolation yields were achieved with the monodisperse-macroporous polymer microspheres carrying trimethoxysilyl and quaternary ammonium functionalities. However, batch DNA isolation performances of polymer based sorbents were significantly lower with respect to the silica microspheres.
Assuntos
DNA/química , DNA/isolamento & purificação , Microesferas , Polímeros/química , Dióxido de Silício/química , Adsorção , Interações Hidrofóbicas e HidrofílicasRESUMO
Monodisperse-porous silica microspheres 5.1µm in size with a bimodal pore-size distribution (including both mesoporous and macroporous compartments) were obtained using a newly developed staged-shape templated hydrolysis and condensation protocol. Synthesized silica microspheres and monodisperse-porous polymer-based microspheres with different functionalities, synthesized by staged-shape template polymerization, were comparatively tested as sorbents for human genomic DNA (hgDNA) isolation in a microfluidic system. Microcolumns with a permeability range of 1.8-8.5×10-13m2 were fabricated by the slurry-packing of silica- or polymer-based microspheres. The monodisperse-porous silica microspheres showed the best performance in hgDNA isolation in an aqueous buffer medium; >2500ng of hgDNA was recovered with an isolation yield of about 50%, using an hgDNA feed concentration of 100ng/µL. Monodisperse-porous silica microspheres were also evaluated as a sorbent for genomic DNA isolation from human whole blood in the microfluidic system; 14ng of hgDNA was obtained from 10µL of whole blood lysate with an isolation yield of 64%. Based on these results, we conclude that monodisperse-porous silica microspheres with a bimodal pore size distribution are a promising sorbent for the isolation of hgDNA in larger amounts and with higher yields compared to the sorbents previously tried in similar microfluidic systems.