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Three-dimensional (3D) printing, an additive manufacturing technique, is increasingly used in the field of tissue engineering. The ability to create complex structures with high precision makes the 3D printing of this material a preferred method for constructing personalized and functional materials. However, the challenge lies in developing affordable and accessible materials with the desired physiochemical and biological properties. In this study, we used eggshell microparticles (ESPs), an example of bioceramic and unconventional biomaterials, to reinforce thermoplastic poly(ε-caprolactone) (PCL) scaffolds via extrusion-based 3D printing. The goal was to conceive a sustainable, affordable, and unique personalized medicine approach. The scaffolds were fabricated with varying concentrations of eggshells, ranging from 0 to 50% (w/w) in the PCL scaffolds. To assess the physicochemical properties, we employed scanning electron microscopy, Fourier-transform infrared spectroscopy, thermogravimetric analysis, differential scanning calorimetry, and X-ray diffraction analysis. Mechanical properties were evaluated through compression testing, and degradation kinetics were studied through accelerated degradation with the remaining mass ranging between 89.4 and 28.3%. In vitro, we evaluated the characteristics of the scaffolds using the MC3T3-E1 preosteoblasts over a 14 day period. In vitro characterization involved the use of the Alamar blue assay, confocal imaging, and real-time quantitative polymerase chain reaction. The results of this study demonstrate the potential of 3D printed biocomposite scaffolds, consisting of thermoplastic PCL reinforced with ESPs, as a promising alternative for bone-graft applications.
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Casca de Ovo , Poliésteres , Impressão Tridimensional , Engenharia Tecidual , Alicerces Teciduais , Alicerces Teciduais/química , Animais , Camundongos , Casca de Ovo/química , Poliésteres/química , Osso e Ossos , Linhagem Celular , Materiais Biocompatíveis/química , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacosRESUMO
Hydrogel-based dressings can effectively heal wounds by providing multiple functions, such as antibacterial, anti-inflammatory, and preangiogenic bioactivities. The ability to spray the dressing is important for the rapid and effective coverage of the wound surface. In this study, we developed a sprayable hydrogel-based wound dressing using naturally derived materials: hyaluronic acid and gelatin. We introduced methacrylate groups (HAMA and GelMA) to these materials to enable controllable photocrosslinking and form a stable hydrogel on the wound surface. To achieve sprayability, we evaluated the concentration of GelMA within a range of 5-15% (w/v) and then incorporated 1% (w/v) HAMA. Additionally, we incorporated calcium peroxide into the hydrogel at concentrations ranging from 0 to 12 mg/mL to provide self-oxygenation and antibacterial properties. The results showed that the composite hydrogels were sprayable and could provide oxygen for up to two weeks. The released oxygen relieved metabolic stress in fibroblasts and reduced cell death under hypoxia in in vitro culture. Furthermore, calcium peroxide added antibacterial properties to the wound dressing. In conclusion, the developed sprayable hydrogel dressing has the potential to be advantageous for wound healing due to its practical and conformable application, as well as its self-oxygenating and antibacterial functions.
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Non-infectious virus-like particles (VLPs) are excellent structures for development of many biomedical applications such as drug delivery systems, vaccine production platforms, and detection techniques for infectious diseases including SARS-CoV-2 VLPs. The characterization of biochemical and biophysical properties of purified VLPs is crucial for development of detection methods and therapeutics. The presence of spike (S) protein in their structure is especially important since S protein induces immunological response. In this study, development of a rapid, low-cost, and easy-to-use technique for both characterization and detection of S protein in the two VLPs, which are SARS-CoV-2 VLPs and HIV-based VLPs was achieved using surface-enhanced Raman spectroscopy (SERS). To analyze and classify datasets of SERS spectra obtained from the VLP groups, machine learning classification techniques including support vector machine (SVM), k-nearest neighbors (kNN), and random forest (RF) were utilized. Among them, the SVM classification algorithm demonstrated the best classification performance for SARS-CoV-2 VLPs and HIV-based VLPs groups with 87.5% and 92.5% accuracy, respectively. This study could be valuable for the rapid characterization of VLPs for the development of novel therapeutics or detection of structural proteins of viruses leading to a variety of infectious diseases.
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COVID-19 , Doenças Transmissíveis , Infecções por HIV , Humanos , SARS-CoV-2 , Análise Espectral Raman , Glicoproteína da Espícula de CoronavírusRESUMO
Hydrogels are often used as biomimetic matrices for tissue regeneration. The source of the hydrogel is of utmost importance, as it affects the physicochemical characteristics and must be carefully selected to stimulate specific cell behaviors. Naturally derived polymeric biomaterials have inherent biological moieties, such as cell binding and protease cleavage sites, and thus can provide a suitable microenvironment for cells. Human-derived matrices can mitigate potential risks associated with the immune response and disease transmission from animal-derived biomaterials. In this article, we developed glycidyl methacrylate-modified human-derived gelatin (hGelGMA) hydrogels for use in tissue engineering applications. By adjusting the glycidyl methacrylate concentration in the reaction mixture, we synthesized hGelGMA with low, medium, and high degrees of modification referred to as hGelGMA-L, hGelGMA-M, and hGelGMA-H, respectively. The amount of polymeric networks in the hydrogels was increased proportionally with the degree of modification. This change has resulted in a decreasing trend in pore size, porosity, and consequent swelling ratio. Similarly, increasing the polymer concentration also exhibited slower enzymatic degradation. On the other hand, increasing the polymer concentration led to an improvement in mechanical properties, where the compressive moduli of hGelGMA-L, hGelGMA-M, and hGelGMA-H hydrogels have changed at 2.9 ± 1.0, 13.7 ± 0.9, and 26.4 ± 2.5 kPa, respectively. The cytocompatibility of hGelGMA was assessed by 3D encapsulation of human-derived cells, including human dermal fibroblasts (HDFs) and human mesenchymal stem cells (hMSCs), in vitro. Regardless of the degree of glycidyl methacrylate modification, the hGelGMA hydrogels preserved the viability of encapsulated cells and supported their growth and proliferation. HDF cells showed a higher metabolic activity in hGelGMA-H, while MSCs exhibited an increased metabolic activity when they were encapsulated in hGelGMA-M or hGelGMA-H. These results showed that photocrosslinkable human-derived gelatin-based hydrogels can be synthesized and their physical properties can be distinctly fine-tuned to different extents as a function of their degrees of modification depending on the needs of the target tissue. Due to its promising physical and biological properties, it is anticipated that hGelGMA can be utilized in a wide spectrum of tissue engineering applications.
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Hidrogéis , Engenharia Tecidual , Animais , Humanos , Engenharia Tecidual/métodos , Hidrogéis/química , Alicerces Teciduais/química , Gelatina/química , Materiais Biocompatíveis/química , PolímerosRESUMO
Cryogel-based scaffolds have attracted great attention in tissue engineering due to their interconnected macroporous structures. However, three-dimensional (3D) printing of cryogels with a high degree of precision and complexity is a challenge, since the synthesis of cryogels occurs under cryogenic conditions. In this study, we demonstrated the fabrication of cryogel-based scaffolds for the first time by using an embedded printing technique. A photo-cross-linkable gelatin methacryloyl (GelMA)-based ink composition, including alginate and photoinitiator, was printed into a nanoclay-based support bath. The layer-by-layer extruded ink was held in complex and overhanging structures with the help of pre-cross-linking of alginate with Ca2+ present in the support bath. The printed 3D structures in the support bath were frozen, and then GelMA was cross-linked at a subzero temperature under UV light. The printed and cross-linked structures were successfully recovered from the support bath with an integrated shape complexity. SEM images showed the formation of a 3D printed scaffold where porous GelMA cryogel was integrated between the cross-linked alginate hydrogels. In addition, they showed excellent shape recovery under uniaxial compression cycles of up to 80% strain. In vitro studies showed that the human fibroblast cells attached to the 3D printed scaffold and displayed spread morphology with a high proliferation rate. The results revealed that the embedded 3D printing technique enables the fabrication of cytocompatible cryogel based scaffolds with desired morphology and mechanical behavior using photo-cross-linkable bioink composition. The properties of the cryogels can be modified by varying the GelMA concentration, whereby various shapes of scaffolds can be fabricated to meet the specific requirements of tissue engineering applications.
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Criogéis , Alicerces Teciduais , Humanos , Criogéis/química , Alicerces Teciduais/química , Engenharia Tecidual/métodos , Impressão Tridimensional , Alginatos/químicaRESUMO
Treating critical-size bone defects with autografts, allografts, or standardized implants is challenging since the healing of the defect area necessitates patient-specific grafts with mechanically and physiologically relevant structures. Three-dimensional (3D) printing using computer-aided design (CAD) is a promising approach for bone tissue engineering applications by producing constructs with customized designs and biomechanical compositions. In this study, we propose 3D printing of personalized and implantable hybrid active scaffolds with a unique architecture and biomaterial composition for critical-size bone defects. The proposed 3D hybrid construct was designed to have a gradient cell-laden poly(ethylene glycol) (PEG) hydrogel, which was surrounded by a porous polycaprolactone (PCL) cage structure to recapitulate the anatomical structure of the defective area. The optimized PCL cage design not only provides improved mechanical properties but also allows the diffusion of nutrients and medium through the scaffold. Three different designs including zigzag, zigzag/spiral, and zigzag/spiral with shifting the zigzag layers were evaluated to find an optimal architecture from a mechanical point of view and permeability that can provide the necessary mechanical strength and oxygen/nutrient diffusion, respectively. Mechanical properties were investigated experimentally and analytically using finite element analysis (FEA), and computational fluid dynamics (CFD) simulation was used to determine the permeability of the structures. A hybrid scaffold was fabricated via 3D printing of the PCL cage structure and a PEG-based bioink comprising a varying number of human bone marrow mesenchymal stem cells (hBMSCs). The gradient bioink was deposited inside the PCL cage through a microcapillary extrusion to generate a mineralized gradient structure. The zigzag/spiral design for the PCL cage was found to be mechanically strong with sufficient and optimum nutrient/gas axial and radial diffusion while the PEG-based hydrogel provided a biocompatible environment for hBMSC viability, differentiation, and mineralization. This study promises the production of personalized constructs for critical-size bone defects by printing different biomaterials and gradient cells with a hybrid design depending on the need for a donor site for implantation.
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Materiais Biocompatíveis , Alicerces Teciduais , Humanos , Alicerces Teciduais/química , Materiais Biocompatíveis/química , Engenharia Tecidual/métodos , Impressão Tridimensional , Hidrogéis/químicaRESUMO
Tissue interfaces include complex gradient structures formed by transitioning of biochemical and mechanical properties in micro-scale. This characteristic allows the communication and synchronistic functioning of two adjacent but distinct tissues. It is particularly challenging to restore the function of these complex structures by transplantation of scaffolds exclusively produced by conventional tissue engineering methods. Three-dimensional (3D) bioprinting technology has opened an unprecedented approach for precise and graded patterning of chemical, biological and mechanical cues in a single construct mimicking natural tissue interfaces. This paper reviews and highlights biochemical and biomechanical design for 3D bioprinting of various tissue interfaces, including cartilage-bone, muscle-tendon, tendon/ligament-bone, skin, and neuro-vascular/muscular interfaces. Future directions and translational challenges are also provided at the end of the paper.
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Bioimpressão , Alicerces Teciduais , Alicerces Teciduais/química , Bioimpressão/métodos , Engenharia Tecidual/métodos , Cartilagem , Tendões , Impressão TridimensionalRESUMO
Bone defect treatment is still a challenge in clinics, and synthetic bone scaffolds with adequate mechanical and biological properties are highly needed. Adequate waste and nutrient exchange of the implanted scaffold with the surrounded tissue is a major concern. Moreover, the risk of mechanical instability in the defect area during regular activity increases as the defect size increases. Thus, scaffolds with better mass transportation and mechanical properties are desired. This study introduces 3D printed polymeric scaffolds with a continuous pattern, ZigZag-Spiral pattern, for bone defects treatments. This pattern has a uniform distribution of pore size, which leads to uniform distribution of wall shear stress which is crucial for uniform differentiation of cells attached to the scaffolds. The mechanical, mass transportation, and biological properties of the 3D printed scaffolds are evaluated. The results show that the presented scaffolds have permeability similar to natural bone and, with the same porosity level, have higher mechanical properties than scaffolds with conventional lay-down patterns 0-90° and 0-45°. Finally, human mesenchymal stem cells are seeded on the scaffolds to determine the effects of geometrical microstructure on cell attachment and morphology. The results show that cells in scaffold with ZigZag-Spiral pattern infilled pores gradually, while the other patterns need more time to fill the pores. Considering mechanical, transportation, and biological properties of the considered patterns, scaffolds with ZigZag-Spiral patterns can mimic the properties of cancellous bones and be a better choice for treatments of bone defects.
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Osso e Ossos , Alicerces Teciduais , Humanos , Porosidade , Impressão Tridimensional , Estresse Mecânico , Engenharia Tecidual/métodos , Alicerces Teciduais/químicaRESUMO
Chronic wounds severely affect 1-2% of the population in developed countries. It has been reported that nearly 6.5 million people in the United States suffer from at least one chronic wound in their lifetime. The treatment of chronic wounds is critical for maintaining the physical and mental well-being of patients and improving their quality of life. There are a host of methods for the treatment of chronic wounds, including debridement, hyperbaric oxygen therapy, ultrasound, and electromagnetic therapies, negative pressure wound therapy, skin grafts, and hydrogel dressings. Among these, hydrogel dressings represent a promising and viable choice because their tunable functional properties, such as biodegradability, adhesivity, and antimicrobial, anti-inflammatory, and pre-angiogenic bioactivities, can accelerate the healing of chronic wounds. This review summarizes the types of chronic wounds, phases of the healing process, and key therapeutic approaches. Hydrogel-based dressings are reviewed for their multifunctional properties and their advantages for the treatment of chronic wounds. Examples of commercially available hydrogel dressings are also provided to demonstrate their effectiveness over other types of wound dressings for chronic wound healing.
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Three-dimensional (3D) bioprinting is an additive manufacturing process in which the combination of biomaterials and living cells, referred to as a bioink, is deposited layer-by-layer to form biologically active 3D tissue constructs. Recent advancements in the field show that the success of this technology requires the development of novel biomaterials or the improvement of existing bioinks. Polyethylene glycol (PEG) is one of the well-known synthetic biomaterials and has been commonly used as a photocrosslinkable bioink for bioprinting; however, other types of cell-friendly crosslinking mechanisms to form PEG hydrogels need to be explored for bioprinting and tissue engineering. In this work, we proposed micro-capillary based bioprinting of a novel molecularly engineered PEG-based bioink that transiently incorporates low molecular weight gelatin (LMWG) fragments. The rheological properties and release profile of the LMWG fragments were characterized, and their presence during hydrogel formation had no effect on the swelling ratio or sol fraction when compared to PEG hydrogels formed without the LMWG fragments. For bioprinting, PEG was first functionalized with cell-adhesive RGD ligands and was then crosslinked using protease-sensitive peptides via a Michael-type addition reaction inside the micro-capillary. The printability was assessed by the analysis of extrudability, shape fidelity, and printing accuracy of the hydrogel filaments after the optimization of the gelation conditions of the PEG-based bioink. The LMWG fragments supplemented into the bioink allowed the extrusion of smooth and uniform cylindrical strands of the hydrogel and improved shape fidelity and printing accuracy. Encapsulated cells in both bioprinted and non-bioprinted PEG-based hydrogels showed high viability and continued to proliferate over time in culture with a well-defined cell morphology depending on the presence of the cell adhesive peptide RGD. The presented micro-capillary based bioprinting process for a novel PEG-based bioink can be promising to construct complex 3D structures with micro-scale range and spatiotemporal variations without using any cytotoxic photoinitiator, UV light, or polymer support.
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Bioimpressão , Materiais Biocompatíveis , Gelatina , Hidrogéis , Impressão Tridimensional , Engenharia Tecidual , Alicerces TeciduaisRESUMO
OBJECTIVES: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) selectively triggers apoptosis in cancer cells, but not in normal cells. Resistance of glioblastoma cells to TRAIL is a major obstacle for successful clinical treatment of TRAIL. Thus, there is an essential requirement for novel approaches to sensitize TRAIL resistance. Silver nanoparticles (AgNPs) are one of the most promising nanomaterials that show immense antitumor potential via targeting various cellular and molecular processes; however, the effects of AgNPs on TRAIL sensitivity in cancer cells remain unclear. Therefore, we hypothesized that TRAIL-conjugated AgNPs (TRAIL-AgNPs) can overcome TRAIL resistance through inducing death receptor activation in glioblastoma cells, but not normal cells. METHODS: In this study, the therapeutic effect of TRAIL-AgNPs is investigated by analyzing the cell viability, caspase activity, and CHK1 gene expression in T98 G TRAIL-Sensitive (TS) and T98 G TRAIL-Resistant (TR) glioblastoma cells. RESULTS: It is found that TRAIL-AgNPs are more toxic compared to TRAIL and AgNPs treatments alone on TR cells. While TRAIL and AgNPs alone do not enhance the caspase activity, conjugation of TRAIL to AgNPs increases the caspase activity in TR cells. Moreover, the TRAIL-AgNPs-treated TR cells show less CHK1 expression compared to the TRAIL treatment. CONCLUSION: These results suggest that TRAIL sensitivity of TR cells can be enhanced by conjugation of TRAIL with AgNPs, which would be a novel therapeutic approach to sensitize TRAIL resistance.
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Sobrevivência Celular/efeitos dos fármacos , Reparo do DNA/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Glioblastoma/metabolismo , Prata/farmacologia , Apoptose/efeitos dos fármacos , Proteínas Reguladoras de Apoptose/genética , Reparo do DNA/genética , Regulação Neoplásica da Expressão Gênica/genética , Glioblastoma/genética , Humanos , Prata/metabolismo , Ligante Indutor de Apoptose Relacionado a TNF/genética , Ligante Indutor de Apoptose Relacionado a TNF/farmacologiaRESUMO
Three-dimensional bioprinting of cell-laden hydrogels in a sacrificial support-bath has recently emerged as a potential solution for fabricating complex biological structures. Physical properties of the support-bath strongly influence the bioprinting process and the outcome of the fabricated constructs. In this study, we reported the application of a composite Pluronic-nanoclay support-bath including calcium ions as the crosslinking agent for bioprinting of cell-laden alginate-based hydrogels. By tuning the rheological properties, a shear-thinning composite support-bath with fast self-recovery behavior was yielded, which allowed continuous printing of complex and large-scale structures. The printed structures were easily and efficiently harvested from the support-bath without disturbing their shape fidelity. Moreover, the results showed that support-bath assisted bioprinting process did not influence the viability of cells encapsulated within hydrogel. This study demonstrates that Pluronic-nanoclay support-bath can be utilized for bioprinting of complex, cell-laden constructs for vascular and other tissue engineering applications.
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Surface-enhanced Raman scattering (SERS)-based single-cell analysis is an emerging approach to obtain molecular level information from molecular dynamics in a living cell. In this study, endosomal biochemical dynamics was investigated based on size and surface chemistry-dependent uptake of gold nanoparticles (AuNPs) on single cells over time using SERS. MDA-MB-231 breast cancer cells were exposed to 13 and 50 nm AuNPs and their polyadenine oligonucleotide-modified forms by controlling the order and combination of AuNPs. The average spectra obtained from 20 single cells were analyzed to study the nature of the biochemical species or processes taking place on the AuNP surfaces. The spectral changes, especially from proteins and lipids of endosomal vesicles, were observed depending on the size, surface chemistry, and combination as well as the duration of the AuNP treatment. The results demonstrate that SERS spectra are sensitive to trace biochemical changes not only the size, surface chemistry, and aggregation status of AuNPs but also the endosomal maturation steps over time, which can be simple and fast way for understanding the AuNP behavior in single cell and useful for the assisting and controlling of AuNP-based gene or drug delivery applications.
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Endossomos/química , Ouro/química , Nanopartículas Metálicas/química , Humanos , Tamanho da Partícula , Análise Espectral Raman , Propriedades de SuperfícieRESUMO
Three-dimensional (3D) spheroid cultures are more realistic tissue mimicking structures for drug discovery studies. However, analysis of 3D spheroid cultures is a challenge task because available techniques are destructive, which results with the loss of biochemical information confined in a spatial arrangement inside of spheroids. In this study, a surface-enhanced Raman scattering (SERS) based non-destructive approach is reported to study 3D cultures. Since the technique uses gold nanoparticles (AuNPs) as SERS substrates, the cells treated with AuNPs are used for the preparation of spheroids. Since SERS spectra originate from molecular species near AuNPs and their aggregates in endolysosomes, the obtained spectral information can provide significant level of information about biomolecular processes taking place in endolysosomes and dependently in cells. The performance of the approach is evaluated by monitoring the spectral changes upon external stimuli with Doxorubicin (Dox) and Paclitaxel (Pac). A layer-by-layer depth-scan SERS analysis of Dox and Pac treated spheroids reveals the spectral changes at around 555â¯cm-1 and 675â¯cm-1 originating from cholesterol and guanine, respectively, compared to control (un-treated) spheroids. Higher spectral variation is observed from the inner to the outer layers of spheroid surface. The results demonstrate that the approach can be used to monitor the intracellular responses, which are in correlation with endolysosomal pathway according to the depth-layers of intact and living 3D spheroids upon external stimuli.
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Ensaios de Seleção de Medicamentos Antitumorais/métodos , Análise Espectral Raman , Esferoides Celulares/efeitos dos fármacos , Análise Discriminante , Doxorrubicina/farmacologia , Células HeLa , Humanos , Paclitaxel/farmacologia , Análise de Componente Principal , Propriedades de SuperfícieRESUMO
Zinc oxide nanoparticles (ZnO) are presented as potential cancer therapeutic agent based on their surface properties. In this study, the most abundant blood proteins, albumin, fibrinogen and apo-transferrin, were covalently bound (c-ZnO NPs) and nonspecifically adsorbed (n-ZnO NPs) onto ZnO NPs to evaluate the role of modification route on protein structure and their effects on glioblastoma cells. The success of modification and structures of proteins on ZnO NPs were characterized with FT-IR. It was found that non-covalent interaction significantly damaged the secondary structure of proteins compared to those covalently attached to the ZnO nanoparticle. The effects of modified ZnO NPs were investigated by evaluating viability, cycle, and death mechanisms of glioblastoma (U373) cells. n-ZnO NPs were found more toxic compared to the pristine and c-ZnO NPs. However, c-ZnO NPs with albumin and apo-transferrin both perturbed the cell cycle function, and decreased the necrotic cell death rate of U373 cells below toxic concentration, suggesting their potential curative effect on glioblastoma cells.
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Proliferação de Células/efeitos dos fármacos , Glioblastoma/tratamento farmacológico , Nanopartículas Metálicas/química , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Glioblastoma/patologia , Humanos , Nanopartículas Metálicas/ultraestrutura , Conformação Proteica/efeitos dos fármacos , Espectroscopia de Infravermelho com Transformada de Fourier , Propriedades de Superfície , Óxido de Zinco/química , Óxido de Zinco/farmacologiaRESUMO
Single cell analysis is an active research area with the hope that cellular process can be deciphered from a single living cell other than a cell population. Surface enhanced Raman scattering (SERS) has been increasingly investigated for single cell analysis with its ability to provide information about real-time dynamics of molecular processes taking place in living cells, especially upon external stimulation, in a contactless, noninvasive, and nondestructive way. In this perspective, the fundamental concepts of single cell-SERS analysis including origin of spectral bands and experimental parameters for spectral reproducibility are summarized along with the recent developments.
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Análise de Célula Única/métodos , Análise Espectral Raman/métodos , Linhagem Celular Tumoral , Ouro/química , Humanos , Nanopartículas Metálicas/química , Reprodutibilidade dos Testes , Prata/químicaRESUMO
Adropin is a peptide hormone that has been implicated in insulin resistance and as a potential regulator of growth. The aim of this study is to determine the effect of calorie restriction on circulating levels of adropin in the MMTV-TGFα breast cancer mouse model and investigate the effects of adropin peptide on the viability of MCF-7 and MDA-231 breast cancer cells in culture. Ten-week-old mice were assigned to either ad libitum-fed (AL), chronic calorie-restricted, or intermittent calorie-restricted groups. Concentrations of serum adropin were measured using an enzyme-linked immunosorbent assay. Results showed an inverse correlation between serum adropin levels and mouse age that was attenuated by calorie restriction. In the AL group the level of adropin was significantly lower at week 50 compared to levels at week 10. However, among the calorie-restricted groups, serum levels of adropin remained high at week 50. The cell-line-specific effects were observed after treatment of cancer cell lines with a series of adropin concentrations (5, 10, 25, 50 ng/mL). Flow cytometry analysis showed that MCF-7 cells entered the early phase of apoptosis after treatment with 50 ng/mL for 24 h. Adropin may be involved in the protective effects that calorie restriction has on breast cancer risk.
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Neoplasias da Mama/tratamento farmacológico , Restrição Calórica/métodos , Proteínas/metabolismo , Fatores Etários , Animais , Apoptose/efeitos dos fármacos , Proteínas Sanguíneas/farmacologia , Peso Corporal , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Feminino , Humanos , Peptídeos e Proteínas de Sinalização Intercelular , Células MCF-7 , Masculino , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Fator de Crescimento Transformador alfa/genéticaRESUMO
Cell lines established from tumors are the most commonly used models in cancer research, and their use in recent years has enabled a greater understanding of the biology of cancer and the means to develop effective treatment strategies. Astroblastomas are uncommon neuroepithelial tumors of glial origin, predominantly affecting young people, mainly teenagers and children, predominantly females. To date, only a single study has reported that astroblastomas contain a large number of neural stem-like cells, which had only a partial proliferation capacity and differentiation. Our objective was to establish an astroblastoma cell line to investigate the presence of astroblastic cells and cancer stem-like cells. The migratory and invasion abilities of the cells were quantified with invasion and migration assays and compared to a glioblastoma cell line. The presence of stem cells was detected with surface-marker analysis by using flow cytometry, and measuring the differentiation ability with a differentiation assay and the self-renewal capacity with a sphere-forming assay. These characteristics may determine whether this novel cell line is a model for astroblastomas that may have stem-cell characteristics. With this novel cell line, scientists can investigate the molecular pathways underlying astroblastomas and develop new therapeutic strategies for patients with these tumors.
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Técnicas de Cultura de Células/métodos , Neoplasias Neuroepiteliomatosas/patologia , Células-Tronco Neoplásicas/citologia , Linhagem Celular Tumoral , Separação Celular/métodos , Humanos , Masculino , Células-Tronco Neoplásicas/fisiologia , Esferoides Celulares/citologia , Esferoides Celulares/fisiologia , Células Tumorais Cultivadas , Adulto JovemRESUMO
The need for new therapeutic approaches in the treatment of challenging diseases such as cancer, which often consists of a highly heterogeneous and complex population of cells, brought up the idea of analyzing single cells. The development of novel techniques to analyze single cells has been intensively studied to fully understand specific alternations inducing abnormalities in cellular function. One of the techniques used for single cell analysis is surface-enhanced Raman spectroscopy (SERS) in which a noble metal nanoparticle is used to enhance Raman scattering. Due to its low toxicity and biocompatibility, gold nanoparticles (AuNPs) are commonly preferred as SERS substrates in single cell analysis. The intracellular uptake, localization and toxicity issues of AuNPs are the critical points for interpretation of data since the obtained SERS signals originate from molecules in close vicinity to AuNPs that are taken up by the cells. In this review, the AuNP-living cell interactions, cellular uptake and toxicity of AuNPs in relation to their physicochemical properties, and surface-enhanced Raman scattering from single cells are discussed.
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Ouro/química , Nanopartículas Metálicas/química , Análise de Célula Única/métodos , Análise Espectral Raman/métodos , Animais , Linhagem Celular , Humanos , Microscopia Eletrônica de TransmissãoRESUMO
Silver nanoparticles (AgNPs) are increasingly used in a variety of applications because of their potential antimicrobial activity and their plasmonic and conductivity properties. In this study, we investigated the source of cytotoxicity, genotoxicity, and reactive oxygen species (ROS) production on human dermal fibroblast and human lung cancer (A549) cell lines upon exposure to AgNP colloidal suspensions prepared with the simplest and most commonly used LeeMeisel method with a variety of reaction times and the concentrations of the reducing agent. The AgNPs synthesized with shorter reaction times were more cytotoxic and genotoxic due to the presence of a few nanometer-sized AgNP seeds. The suspensions prepared with an increased citrate concentration were not cytotoxic, but they induced more ROS generation on A549 cells due to the high citrate concentration. The genotoxicity of the suspension decreased significantly at the higher citrate concentrations. The analysis of both transmission electron microscopy images from the dried droplet areas of the colloidal suspensions and toxicity data indicated that the AgNP seeds were the major source of toxicity. The completion of the nucleation step and the formation of larger AgNPs effectively decreased the toxicity.