RESUMO
Progress in medical sciences aims for tailored therapy of civilization diseases like diabetes. Preclinical screening of new medicines superior to insulin should include the verification of their affinity to the membrane receptors naturally stimulated by this hormone: insulin receptor isoforms A and B and insulin-like growth factor receptor. Considering that the affinity constants obtained using different experimental conditions are incomparable, it is essential to develop a robust and reliable method to analyze these interactions. The versatile SPR platform developed in this study enables the evaluation of the bioactivity of hypoglycaemic molecules. Thanks to the comprehensive characterization of miscellaneous aspects of the analytical platform, including the design of the SPR biosensor receptor layer, ensuring interaction specificity, as well as the quality control of the standards used (human insulin, HI; long-acting insulin analog: glargine, Gla), the feasibility of the method of equilibrium and kinetic constants determination for insulin-like targets was confirmed. SPR assays constructed in the direct format using IR-A, IR-B, and IGF1-R receptor proteins show high sensitivities and low detection limits towards insulin and glargine detection in the range of 18.3-53.3 nM with no signs of mass transport limitations. The improved analytical performance and stability of SPR biosensors favor the acquisition of good-quality kinetic data, while preservation of receptors activity after binding to long-chain carboxymethyldextran, combined with spontaneous regeneration, results in stability and long shelf life of the biosensor, which makes it useful for label-free insulin analogs biosensing and thus extensive screening in diabetic drugs discovery.
Assuntos
Ensaios de Triagem em Larga Escala , Hipoglicemiantes , Receptor de Insulina , Ressonância de Plasmônio de Superfície , Humanos , Hipoglicemiantes/química , Ressonância de Plasmônio de Superfície/métodos , Receptor de Insulina/metabolismo , Ensaios de Triagem em Larga Escala/métodos , Insulina Glargina/química , Técnicas Biossensoriais/métodos , Insulina/metabolismo , Insulina/análise , Receptor IGF Tipo 1/metabolismoRESUMO
The article presents magnetoliposomes as potential carriers of doxorubicin. The magnetic properties of nanoparticles embedded in liposomes enable the targeting of drug-loaded carriers to cancer cells and subsequent release of their payload using an external alternating magnetic field as a trigger. The cytotoxicity of empty and doxorubicin-loaded magnetoliposomes in the absence and after exposure to magnetic field was evaluated in cancerous and normal breast cells. The characteristic shows the carrier with size distribution <130â nm, slightly negative zeta potential and polydispersity index <0.2. Doxorubicin was encapsulated in magnetoliposomes with an efficiency of 31 % and released in the presence of an alternating magnetic field at 50 %. Magnetoliposomes with drug provided high cytotoxic effect on tumor cells and low cytotoxic effect on normal cells. The research conducted in this article may indicate the potential application of the studied magnetoliposomes in release of drugs under the influence of magnetic field in cancer cells.
Assuntos
Antineoplásicos , Doxorrubicina , Preparações de Ação Retardada/farmacologia , Doxorrubicina/farmacologia , Lipossomos , Campos Magnéticos , Portadores de FármacosRESUMO
Pythium oligandrum, strain M1, is a soil oomycete successfully used as a biological control agent (BCA), protecting plants against fungal, yeast, and oomycete pathogens through mycoparasitism and elicitor-dependent plant priming. The not yet described Pythium strains, X42 and 00X48, have shown potential as BCAs given the high activity of their secreted proteases, endoglycosidases, and tryptamine. Here, Solanum lycopersicum L. cv. Micro-Tom seeds were coated with Pythium strains, and seedlings were exposed to fungal pathogens, either Alternaria brassicicola or Verticillium albo-atrum. The effects of both infection and seed-coating on plant metabolism were assessed by determining the activity and isoforms of antioxidant enzymes and endoglycosidases and the content of tryptamine, amino acids, and heat shock proteins. Dual culture competition testing and microscopy analysis confirmed mycoparasitism in all three Pythium strains. In turn, seed treatment significantly increased the total free amino acid content, changing their abundance in both non-infected and infected plants. In response to pathogens, plant Hsp70 and Hsp90 isoform levels also varied among Pythium strains, most likely as a strategy for priming the plant against infection. Overall, our results show in vitro mycoparasitism between Pythium strains and fungal pathogens and in planta involvement of heat shock proteins in priming.
RESUMO
A broad family of two-dimensional (2D) materials - carbides, nitrides, and carbonitrides of early transition metals, called MXenes, became a newcomer in the flatland at the turn of 2010 and 2011 (over ten years ago). Their unique physicochemical properties made them attractive for many applications, highly boosting the development of various fields, including biotechnological. However, MXenes' functional features that impact their bioactivity and toxicity are still not fully well understood. This study discusses the essentials for MXenes's surface modifications toward their application in modern biotechnology and nanomedicine. We survey modification strategies in context of cytotoxicity, biocompatibility, and most prospective applications ready to implement in medical practice. We put the discussion on the material-structure-chemistry-property relationship into perspective and concentrate on overarching challenges regarding incorporating MXenes into nanostructured organic/inorganic bioactive architectures. It is another emerging group of materials that are interesting from the biomedical point of view as well. Finally, we present an influential outlook on the growing demand for future research in this field.
Assuntos
Nanopartículas/química , Elementos de Transição/química , Anti-Infecciosos/farmacologia , Materiais Biocompatíveis/química , Materiais Biocompatíveis/toxicidade , Sobrevivência Celular , Estabilidade de Medicamentos , Humanos , Nanopartículas/toxicidade , Fotoquimioterapia/métodos , Relação Estrutura-Atividade , Propriedades de Superfície , Condutividade Térmica , Elementos de Transição/toxicidadeRESUMO
Our skin is continuously exposed to different amphiphilic substances capable of interaction with its lipids and proteins. We describe the effect of a saponin-rich soapwort extract and of four commonly employed synthetic surfactants: sodium lauryl sulfate (SLS), sodium laureth sulfate (SLES), ammonium lauryl sulfate (ALS), cocamidopropyl betaine (CAPB) on different human skin models. Two human skin cell lines were employed: normal keratinocytes (HaCaT) and human melanoma cells (A375). The liposomes consisting of a dipalmitoylphosphatidylcholine/cholesterol mixture in a molar ratio of 7:3, mimicking the cell membrane of keratinocytes and melanoma cells were employed as the second model. Using dynamic light scattering (DLS), the particle size distribution of liposomes was analyzed before and after contact with the tested (bio)surfactants. The results, supplemented by the protein solubilization tests (albumin denaturation test, zein test) and oil emulsification capacity (using olive oil and engine oil), showed that the soapwort extract affects the skin models to a clearly different extent than any of the tested synthetic surfactants. Its protein and lipid solubilizing potential are much smaller than for the three anionic surfactants (SLS, ALS, SLES). In terms of protein solubilization potential, the soapwort extract is comparable to CAPB, which, however, is much harsher to lipids.
Assuntos
Biomimética/métodos , Extratos Vegetais/química , Saponaria/química , Pele/efeitos dos fármacos , Tensoativos/química , 1,2-Dipalmitoilfosfatidilcolina/química , Betaína/análogos & derivados , Betaína/química , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Colesterol/química , Emulsificantes/química , Humanos , Concentração de Íons de Hidrogênio , Queratinócitos/efeitos dos fármacos , Lipossomos/química , Modelos Biológicos , Tamanho da Partícula , Saponinas/química , Dodecilsulfato de Sódio/análogos & derivados , Dodecilsulfato de Sódio/química , Triterpenos/química , Zeína/químicaRESUMO
Malignant melanoma is an emerging problem worldwide due to the high degree of lethalness. Its aggressiveness and the ability to metastasize along with the heterogeneity at the molecular and cellular levels, limit the overall therapeutic efficacy. Despite significant advances in melanoma treatment over the last decade, there is still a need for improved therapeutic modalities. Thus, we demonstrate here a combinatorial approach that targets multiple independent therapeutic pathways, in which polymeric micelles (PMs) were used as efficacious colloidal nanocarriers loaded with both daunorubicin (DRB) as a cytotoxic drug and IR-768 as a photosensitizer. This afforded the dual drug loaded delivery system IR-768 + DRB in PMs. The fabricated mPEG-b-PLGA micelles (hydrodynamic diameters ≈ 25 nm) had a relatively narrow size distribution (PdI > ca. 0.3) with uniform spherical shapes. CLSM study showed that mPEG-b-PLGA micelles were uptaken by mitochondria, which further contributed to excellent singlet oxygen generation capacity for PDT in A375 melanoma cells. Furthermore, the PMs were efficiently internalized by tested cells through endocytosis, resulting in much higher cellular uptake comparing to the free drug. As a result of these properties, IR-768 + DRB in PMs exhibited very potent and synergistically enhanced anticancer activity against A375 cells. Additionally, this combination approach allowed to reduce drug doses and provided low side effects towards normal HaCaT. This study indicates excellent properties of mPEG-b-PLGA micelles resulting in great therapeutic potential possessed by the developed nanoscale drug delivery system for combined chemo-photodynamic therapy of melanoma.
Assuntos
Antineoplásicos/química , Daunorrubicina/química , Melanoma/terapia , Nanocápsulas/química , Fármacos Fotossensibilizantes/química , Poliésteres/química , Polietilenoglicóis/química , Neoplasias Cutâneas/terapia , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Permeabilidade da Membrana Celular , Terapia Combinada , Daunorrubicina/farmacologia , Relação Dose-Resposta a Droga , Composição de Medicamentos , Liberação Controlada de Fármacos , Humanos , Micelas , Fotoquimioterapia , Oxigênio Singlete/metabolismo , Melanoma Maligno CutâneoRESUMO
Current trends in the field of MXenes emphasize the importance of controlling their surface features for successful application in biotechnological areas. The ability to stabilize the surface properties of MXenes has been demonstrated here through surface charge engineering. It was thus determined how changing the surface charges of two-dimensional (2D) Ti3C2 MXene phase flakes using cationic polymeric poly-L-lysine (PLL) molecules affects the colloidal and biological properties of the resulting hybrid 2D nanomaterial. Electrostatic adsorption of PLL on the surface of delaminated 2D Ti3C2 flakes occurs efficiently, leads to changing an MXene's negative surface charge toward a positive value, which can also be effectively managed through pH changes. Analysis of bioactive properties revealed additional antibacterial functionality of the developed 2D Ti3C2/PLL MXene flakes concerning Escherichia. coli Gram-negative bacteria cells. A reduction of two orders of magnitude of viable cells was achieved at a concentration of 200 mg L-1. The in vitro analysis also showed lowered toxicity in the concentration range up to 375 mg L-1. The presented study demonstrates a feasible approach to control surface properties of 2D Ti3C2 MXene flakes through surface charge engineering which was also verified in vitro for usage in biotechnology or nanomedicine applications.
RESUMO
The past few years have seen significant developments in the chemistry and potential biological applications of 2D materials. This review focuses on recent advances in the biotechnological and biomedical applications of MXenes, which are 2D carbides, nitrides, and carbonitrides of transition metals. Nanomaterials based on MXenes can be used as therapeutics for anticancer treatment, in photothermal therapy as drug delivery platforms, or as nanodrugs without any additional modification. Furthermore, we discuss the potential use of these materials in biosensing and bioimaging, including magnetic resonance and photoacoustic imaging techniques. Finally, we present the most significant examples of the use of MXenes as efficient agents for environmental and antimicrobial treatments, as well as a brief discussion of their future prospects and challenges.
Assuntos
Técnicas Biossensoriais/métodos , Diagnóstico por Imagem/métodos , Neoplasias/terapia , Elementos de Transição/química , Animais , Biotecnologia/instrumentação , Biotecnologia/métodos , Humanos , Nanomedicina , Nanoestruturas/química , Nanoestruturas/uso terapêutico , Técnicas Fotoacústicas , Terapia Fototérmica , Elementos de Transição/uso terapêuticoRESUMO
In recent years, colistin has been the drug of choice for treatment of nosocomial infections, especially in bloodstream infections, lower respiratory tract infections, or urinary tract infections. In this study, 65 multidrug-resistant Klebsiella pneumoniae isolated from different clinical samples were included. Minimum inhibitory concentration (MIC) of colistin was detected by broth microdilution method in three different ways. For selected K. pneumoniae strains, eazyplex SuperBug mcr-1 test was performed. This test detects mcr-1 gene, which encodes a colistin-resistance determinant. Most of the analyzed K. pneumoniae strains were resistant to colistin in all applied methods. The exception was two strains, where MIC of colistin was 2 mg/L in SensiTest Colistin and MIC-Strip Colistin tests. In MIC COL test, MIC for these strains was 4 mg/L. All analyzed strains produced extended-spectrum beta-lactamases and 11 (16.9%) metallo-beta-lactamases. Eleven (16.9%) K. pneumoniae strains were resistant to all antibiotics, whereas 17 (26.1%) were susceptible to only one drug. Colistin MIC values varied from 2 to >64 mg/L in MIC-Strip Colistin test; from 2 to >16 mg/L in SensiTest Colistin and from 4 to >16 mg/L in MIC COL test. None of the analyzed K. pneumoniae strains carried mcr-1 gene. Data of this work suggest that resistance to colistin emerged among multidrug-resistant K. pneumoniae strains. The tests allowed for reliable estimation of susceptibility to colistin and could be used in microbiological diagnostics.
Assuntos
Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Colistina/farmacologia , Farmacorresistência Bacteriana Múltipla/genética , Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/efeitos dos fármacos , Humanos , Klebsiella pneumoniae/enzimologia , Klebsiella pneumoniae/genética , Testes de Sensibilidade Microbiana , Fenótipo , Polônia , beta-Lactamases/genéticaRESUMO
The surface organic ligands have profound effect on modulation of different physicochemical parameters as well as toxicological profile of semiconductor nanocrystals (NCs). Zinc oxide (ZnO) is one of the most versatile semiconductor material with multifarious potential applications and systematic approach to in-depth understand the interplay between ZnO NCs surface chemistry along with physicochemical properties and their nano-specific toxicity is indispensable for development of ZnO NCs-based devices and biomedical applications. To this end, we have used recently developed the one-pot self-supporting organometallic (OSSOM) approach as a model platform to synthesize a series of ZnO NCs coated with three different alkoxyacetate ligands with varying the ether tail length which simultaneously act as miniPEG prototypes. The ligand coating influence on ZnO NCs physicochemical properties including the inorganic core size, the hydrodynamic diameter, surface charge, photoluminescence (quantum yield and decay time) and ZnO NCs biological activity toward lung cells was thoroughly investigated. The resulting ZnO NCs with average core diameter of 4-5 nm and the hydrodynamic diameter of 8-13 nm exhibit high photoluminescence quantum yield reaching 33% and a dramatic slowing down of charge recombination up to 2.4 µs, which is virtually unaffected by the ligand's character. Nano-specific ZnO NCs-induced cytotoxicity was tested using MTT assay with normal (MRC-5) and cancer (A549) human lung cell lines. Noticeably, no negative effect has been observed up to the NCs concentration of 10 µg/mL and essentially very low negative toxicological impact could be noticed at higher concentrations. In the latter case, the MTT data analysis indicate that there is a subtle interconnection between inorganic core-organic shell dimensions and toxicological profile of ZnO NCs (strikingly, the NCs coated by the carboxylate bearing a medium ether chain length exhibit the lowest toxicity level). The results demonstrate that, when fully optimized, our organometallic self-supporting approach can be a highly promising method to obtain high-quality and bio-stable ligand-coated ZnO NCs.
RESUMO
Magnetoliposomes are promising candidates for the development of selective drug delivery systems in the treatment of cancer. Those nanosystems were tested as carriers of a strong chemotherapeutic agent, doxorubicin, which is used against breast cancer. Herein, the magnetic properties of hydrophobic iron oxide nanoparticles located exclusively in the lipid bilayer were used to release this drug from the magnetoliposomes. The cytotoxic activity of the nanostructures against the normal and cancer cell lines was determined on the basis of cells viability measurement after incubation with different concentrations of these nanomaterials. In the same way, the effectiveness of killing cancer cells in combination with exposure to magnetic field was also evaluated. These experiments confirmed that the nanostructures composed of liposomes and magnetic nanoparticles are not cytotoxic. However, magnetoliposomes loaded with doxorubicin were effective and selective in reducing the viability of human breast tumor cell lines. In this paper, we demonstrated the promising application of the studied magnetoliposomes as carriers of doxorubicin released under the influence of magnetic field in tumor cells.
Assuntos
Antibióticos Antineoplásicos/administração & dosagem , Neoplasias da Mama/tratamento farmacológico , Preparações de Ação Retardada/química , Doxorrubicina/administração & dosagem , Nanopartículas de Magnetita/química , Antibióticos Antineoplásicos/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Doxorrubicina/farmacologia , Feminino , Humanos , Lipossomos/química , Células MCF-7 , Campos MagnéticosRESUMO
Taste sensing is of great importance in both the pharmaceutical and foodstuff industries, and is currently mainly based on human sensory evaluation. Many approaches based on chemical sensors have been proposed, leading to the development of various electronic tongue systems. However, this approach is limited by the applied recognition methods, which do not consider natural receptors. Biorecognition elements such as taste receptor proteins or whole cells can be involved in the development of taste sensing biosensors usually equipped with various electrochemical transducers. Here, we propose a new approach: intestinal secretin tumor cell line (STC-1) chemosensory cells were applied for taste recognition, and their taste-specific cellular response was decoded from ion chromatographic fingerprints with the use of multivariate data processing by partial least squares discriminant analysis (PLS-DA). This approach could be useful for the development of various non-invasive taste sensing assays, as well as for studying taste transduction mechanisms in vitro.
Assuntos
Técnicas Biossensoriais/métodos , Cromatografia/métodos , Cromatografia por Troca Iônica/métodos , Humanos , Paladar/fisiologiaRESUMO
Photothermal therapy (PTT) has shown significant potential for anti-cancer modality. In this report, according to our best knowledge, we explore for the first time Ti2C-based MXene as a novel, highly efficient and selective agent for photothermal therapy (PTT). Ti2C superficially modified with PEG was obtained from the layered, commercially available Ti2AlC MAX phase in the process of etching aluminum layers using concentrated HF, and characterized by scanning electron microscopy (SEM), high-resolution transmission electron microscopy (HREM) as well as X-ray photoelectron spectroscopy for chemical analysis (ESCA-XPS). The PEG-coated Ti2C flakes showed a satisfactory photothermal conversion efficacy (PTCE) and good biocompatibility in wide range of the tested concentrations. Through in vitro studies, the PEG-modified Ti2C demonstrated notable NIR-induced ability to cancerous cells' ablation with minimal impact on non-malignant cells up to the concentration of 37.5⯵gâ¯mL-1. The applied doses of Ti2C_PEG in our work were even 24 times lower comparing other MXene-based photothermal agents. This work is expected to expand the utility of 2D MXenes to biomedical applications through the development of entirely novel agents for photothermal therapy. This work is expected to expand the utility of 2D MXenes to biomedical applications through the development of entirely novel agents for photothermal therapy.
Assuntos
Hipertermia Induzida , Fototerapia , Titânio/química , Morte Celular , Linhagem Celular Tumoral , Humanos , Espectroscopia Fotoeletrônica , Polietilenoglicóis/química , Espécies Reativas de Oxigênio/metabolismo , TemperaturaRESUMO
This up-to-date review summarizes the design and current fabrication strategies that have been employed in the area of mono- and multifunctional colloidal nanoparticles - nanocarriers well suited for photodynamic therapy (PDT) and diagnostic purposes. Rationally engineered photosensitizer (PS)-loaded nanoparticles may be achieved via either noncovalent (i.e., self-aggregation, interfacial deposition, interfacial polymerization, or core-shell entrapment along with physical adsorption) or covalent (chemical immobilization or conjugation) processes. These PS loading approaches should provide chemical and physical stability to PS payloads. Their hydrophilic surfaces, capable of appreciable surface interactions with biological systems, can be further modified using functional groups (stealth effect) to achieve prolonged circulation in the body after administration and/or grafted by targeting agents (such as ligands, which bind to specific receptors uniquely expressed on the cell surface) or stimuli (e.g., pH, temperature, and light)-responsive moieties to improve their action and targeting efficiency. These attempts may in principle permit efficacious PDT, combination therapies, molecular diagnosis, andâ¯-â¯in the case of nanotheranostics - simultaneous monitoring and treatment. Nanophotosensitizers (nano-PSs) should possess appropriate morphologies, sizes, unimodal distributions and surface processes to be successfully delivered to the place of action after systemic administration and should be accumulated in certain tumors by passive and/or active targeting. Additionally, physically facilitating drug delivery systems emerge as a promising approach to enhancing drug delivery, especially for the non-invasive treatment of deep-seated malignant tissues. Recent advances in nano-PSs are scrutinized, with an emphasis on design principles, via the promising use of colloid chemistry and nanotechnology.
Assuntos
Imagem Molecular , Nanopartículas/química , Fotoquimioterapia , Fármacos Fotossensibilizantes/química , Coloides/química , HumanosRESUMO
The unique physicochemical properties and biocompatibility of zinc oxide nanocrystals (ZnO NCs) are strongly dependent on the nanocrystal/ligand interface, which is largely determined by synthetic procedures. Stable ZnO NCs coated with a densely packed shell of 2-(2-methoxyethoxy)acetate ligands, which act as miniPEG prototypes, with average core size and hydrodynamic diameter of 4-5 and about 12â nm, respectively, were prepared by an organometallic self-supporting approach, fully characterized, and used as a model system for biological studies. The ZnO NCs from the one-pot, self-supporting organometallic procedure exhibit unique physicochemical properties such as relatively high quantum yield (up to 28 %), ultralong photoluminescence decay (up to 2.1â µs), and EPR silence under standard conditions. The cytotoxicity of the resulting ZnO NCs toward normal (MRC-5) and cancer (A549) human lung cell lines was tested by MTT assay, which demonstrated that these brightly luminescent, quantum-sized ZnO NCs have a low negative impact on mammalian cell lines. These results substantiate that the self-supporting organometallic approach is a highly promising method to obtain high-quality, nontoxic, ligand-coated ZnO NCs with prospective biomedical applications.
Assuntos
Nanopartículas Metálicas/química , Óxido de Zinco/toxicidade , Animais , Humanos , Ligantes , Luminescência , Pulmão/citologia , Pulmão/efeitos dos fármacos , Estudos Prospectivos , Óxido de Zinco/químicaRESUMO
In recent years photodynamic therapy (PDT) has received widespread attention in cancer treatment due to its smaller surgical trauma, better selectivity towards tumor cells, reduced side effects and possibility of repeatable treatment. Since cancer is the second cause of death worldwide, scientists constantly seek for new potential therapeutic agents including nanotechnology-based photosensitizers used in PDT. The new-designed nanostructures must be carefully studied and well characterized what require analytically useful and powerful tools that enable real progress in nanoscience development. This review describes the current status of PDT investigations using microfluidic Lab-on-a-Chip systems, including recent developments of nanoparticle-based PDT agents, their combinations with different drugs, designs and examples of in vitro applications. This review mainly lays emphasis on biological evaluation of FDA approved photosensitizing agents as well as newly designed nanophotosensitizers. It also highlights the analytical performances of various microfluidic Lab-on-a-chip systems for PDT efficacy analysis on 3D culture and discusses microsystems designs in detail.
Assuntos
Técnicas Biossensoriais/instrumentação , Dispositivos Lab-On-A-Chip , Neoplasias/tratamento farmacológico , Fotoquimioterapia , Fármacos Fotossensibilizantes/uso terapêutico , Animais , Técnicas Biossensoriais/métodos , Técnicas de Cultura de Células/instrumentação , Técnicas de Cultura de Células/métodos , Desenho de Equipamento , Humanos , Nanoestruturas/química , Nanoestruturas/uso terapêutico , Fotoquimioterapia/instrumentação , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/químicaRESUMO
The purpose of this paper is to present a fully integrated microchip for the evaluation of PDT procedures efficiency on 3D lung spheroid cultures. Human lung carcinoma A549 and non-malignant MRC-5 spheroids were utilized as culture models. Spheroid viability was evaluated 24 h after PDT treatment, in which 5-aminolevulinic acid (ALA) had been used as a precursor of a photosensitizer (protoporphyrin IX - PpIX). Moreover, spheroid viability over a long-term (10-day) culture was also examined. We showed that the proposed PDT treatment was toxic only for cancer spheroids. This could be because of a much-favoured enzymatic conversion of ALA to PpIX in cancer as opposed normal cells. Moreover, we showed that to obtain high effectiveness of ALA-PDT on lung cancer spheroids additional time of spheroid after light exposure was required. It was found that PDT had been effective 5 days after PDT treatment with 3 mM ALA. To the best of our knowledge this has been the first presentation of such research performed on a 3D lung spheroids culture in a microfluidic system.
Assuntos
Ácido Aminolevulínico/química , Dispositivos Lab-On-A-Chip , Fotoquimioterapia , Fármacos Fotossensibilizantes/química , Protoporfirinas/química , Esferoides Celulares/efeitos dos fármacos , Células A549 , Humanos , Neoplasias Pulmonares/tratamento farmacológicoRESUMO
Lab-on-a-chip systems are increasingly used as tools for cultures and investigation of cardiac cells. In this article, we present how the geometry of microsystems and microenvironmental conditions (static and perfusion) influence the proliferation, morphology, and alignment of cardiac cells (rat cardiomyoblasts-H9C2). Additionally, studies of cell growth after incubation with verapamil hydrochloride were performed. For this purpose, poly(dimethylsiloxane) (PDMS)/glass microfluidic systems with three different geometries of microchambers (a circular chamber, a longitudinal channel, and three parallel microchannels separated by two rows of micropillars) were prepared. It was found that static conditions did not enhance the growth of H9C2 cells in the microsystems. On the contrary, perfusion conditions had an influence on division, morphology, and the arrangement of the cells. The highest number of cells, their parallel orientation, and their elongated morphology were obtained in the longitudinal microchannel. It showed that this kind of microsystem can be used to understand processes in heart tissue in detail and to test newly developed compounds applied in the treatment of cardiac diseases.
Assuntos
Proliferação de Células , Dispositivos Lab-On-A-Chip , Microfluídica/métodos , Miócitos Cardíacos/fisiologia , Animais , Técnicas de Cultura de Células/instrumentação , Técnicas de Cultura de Células/métodos , Linhagem Celular , Microfluídica/instrumentação , Miócitos Cardíacos/citologia , RatosRESUMO
This paper presents a comparison and evaluation of cardiac cell proliferation on poly(l-lactic acid) (PLLA) and polyurethane (PU) nanofibrous mats fabricated by solution blow spinning (SBS). Three different cardiac cell lines: rat cardiomyoblasts (H9C2 line), human (HCM) and rat cardiomyocytes (RCM) were used for experiments. Cell morphology, orientation and proliferation were investigated on non-modified and protein-modified (fibronectin, collagen, gelatin, laminin, poly-l-lysine) surfaces of both types of nanofibers. Obtained results of cell culture on nanofibers surfaces were compared to the results of cell culture on polystyrene (PS) surfaces modified in the same way. The results indicated that in most cases polymeric nanofibers (PLLA and PU) are better substrates for cardiac cell culture than PS surfaces. All types of investigated cells, cultured on nanofibers (PLLA and PU), had more elongated shape than cells cultured on PS surfaces. Moreover, cells were arranged in parallel to each other, according to fibers orientation. Additionally, it was shown that the protein modifications of investigated surfaces influenced on cell proliferation. Therefore, we suggest that the cardiac cell culture on nanofibrous mats fabricated by SBS could be more advanced experimental in vitro model for studies on the effect of various cardiac drugs than traditional culture on PS surface.
Assuntos
Técnicas de Cultura de Células/métodos , Mioblastos Cardíacos/metabolismo , Miócitos Cardíacos/metabolismo , Nanofibras/química , Poliésteres/química , Poliuretanos/química , Animais , Linhagem Celular , Camundongos , Mioblastos Cardíacos/citologia , Miócitos Cardíacos/citologia , RatosRESUMO
INTRODUCTION: Soft tissue mesenchymal tumours are a common occurrence in surgical practice with particular predilection for the extremities. Approximately 1 in 100 soft tissue tumours are found to be sarcomas. The main concern is to exclude any evidence of malignancy. Both imaging studies and a detailed histological analysis is required to ensure that a diagnosis of a high-grade tumour is not missed. PRESENTATION OF CASE: Here we present a 38-year-old previously fit and well gentleman with a slowly growing lump in the upper aspect of his abdomen over the previous year being completely asymptomatic from it. He underwent ultrasound, computed tomography and magnetic resonance imaging of the lump. He underwent ultrasound guided biopsy with eventual wide local excision of the lump for a complete histological assessment. This was noted to be a soft tissue mesenchymal tumour. DISCUSSION: We highlight the importance of review of the literature and the use of markers that enable histopathologist reach an eventual diagnosis. Mesenchymal tissue during development differentiates into fat, skeletal muscle, peripheral nerves, blood vessels and fibrous tissue. Thereby any of these components may give rise to a tumour. In the majority of cases, the patient is asymptomatic unless there is invasion of nerve sheath or the effects of mass effect. CONCLUSION: Our case is unique due to location of the tumour and its immunohistochemistry findings which required frequent and extensive discussion at our national sarcoma soft tissue meeting. The importance of surgeons working with histopathologists was also highlighted in our case.