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1.
Molecules ; 28(17)2023 Aug 25.
Artigo em Inglês | MEDLINE | ID: mdl-37687081

RESUMO

Graphene oxide (GO) is an oxidized derivative of graphene. So far, GO has mostly been studied as a drug delivery method rather than a standalone drug for treating cancers like glioblastoma or cervical cancer. However, we propose a promising new approach-using GO as a sensitizer for cisplatin chemotherapy. Here, we analyze the effects of triple GO pretreatment, followed by cisplatin treatment, on cancerous cell lines U87 and HeLa, as well as the noncancerous cell line HS-5, through morphology analysis, viability assay, flow cytometry, and LDH release assay. The viability assay results showed that GO treatment made U87 and HeLa cells more responsive to cisplatin, leading to a significant reduction in cell viability to 40% and 72%, respectively, without affecting HS-5 cells viability, while the Annexin V/Propidium iodine assay showed that GO pretreatment did not cause a change in live cells in all three examined cell lines, while GO-pretreated HeLa cells treated with cisplatin showed significant decrease around two times compared to cells treated with cisplatin standalone. The U87 cell line showed a significant increase in LDH release, approximately 2.5 times higher than non-GO-pretreated cells. However, GO pretreatment did not result in LDH release in noncancerous HS-5 cells. It appears that this phenomenon underlays GO's ability to puncture the cell membrane of cancerous cells depending on its surface properties without harming noncancerous cells.


Assuntos
Antineoplásicos , Glioblastoma , Grafite , Neoplasias do Colo do Útero , Humanos , Feminino , Cisplatino/farmacologia , Grafite/farmacologia , Neoplasias do Colo do Útero/tratamento farmacológico , Glioblastoma/tratamento farmacológico , Células HeLa , Antineoplásicos/farmacologia
2.
Small ; 18(2): e2104971, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-34802179

RESUMO

The use of injectable biomaterials for cell delivery is a rapidly expanding field which may revolutionize the medical treatments by making them less invasive. However, creating desirable cell carriers poses significant challenges to the clinical implementation of cell-based therapeutics. At the same time, no method has been developed to produce injectable microscaffolds (MSs) from electrospun materials. Here the fabrication of injectable electrospun nanofibers is reported on, which retain their fibrous structure to mimic the extracellular matrix. The laser-assisted micro-scaffold fabrication has produced tens of thousands of MSs in a short time. An efficient attachment of cells to the surface and their proliferation is observed, creating cell-populated MSs. The cytocompatibility assays proved their biocompatibility, safety, and potential as cell carriers. Ex vivo results with the use of bone and cartilage tissues proved that NaOH hydrolyzed and chitosan functionalized MSs are compatible with living tissues and readily populated with cells. Injectability studies of MSs showed a high injectability rate, while at the same time, the force needed to eject the load is no higher than 25 N. In the future, the produced MSs may be studied more in-depth as cell carriers in minimally invasive cell therapies and 3D bioprinting applications.


Assuntos
Nanofibras , Materiais Biocompatíveis/química , Matriz Extracelular/química , Lasers , Nanofibras/química , Engenharia Tecidual/métodos , Alicerces Teciduais/química
3.
Int J Mol Sci ; 22(2)2021 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-33419226

RESUMO

The development of nanotechnology based on graphene and its derivatives has aroused great scientific interest because of their unusual properties. Graphene (GN) and its derivatives, such as reduced graphene oxide (rGO), exhibit antitumor effects on glioblastoma multiforme (GBM) cells in vitro. The antitumor activity of rGO with different contents of oxygen-containing functional groups and GN was compared. Using FTIR (fourier transform infrared) analysis, the content of individual functional groups (GN/exfoliation (ExF), rGO/thermal (Term), rGO/ammonium thiosulphate (ATS), and rGO/ thiourea dioxide (TUD)) was determined. Cell membrane damage, as well as changes in the cell membrane potential, was analyzed. Additionally, the gene expression of voltage-dependent ion channels (clcn3, clcn6, cacna1b, cacna1d, nalcn, kcne4, kcnj10, and kcnb1) and extracellular receptors was determined. A reduction in the potential of the U87 glioma cell membrane was observed after treatment with rGO/ATS and rGO/TUD flakes. Moreover, it was also demonstrated that major changes in the expression of voltage-dependent ion channel genes were observed in clcn3, nalcn, and kcne4 after treatment with rGO/ATS and rGO/TUD flakes. Furthermore, the GN/ExF, rGO/ATS, and rGO/TUD flakes significantly reduced the expression of extracellular receptors (uPar, CD105) in U87 glioblastoma cells. In conclusion, the cytotoxic mechanism of rGO flakes may depend on the presence and types of oxygen-containing functional groups, which are more abundant in rGO compared to GN.


Assuntos
Canais de Cloreto/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Glioblastoma/genética , Grafite/farmacologia , Canais Iônicos/genética , Proteínas de Membrana/genética , Canais de Potássio de Abertura Dependente da Tensão da Membrana/genética , Receptores de Superfície Celular/genética , Linhagem Celular Tumoral , Células , Canais de Cloreto/metabolismo , Glioblastoma/metabolismo , Glioblastoma/patologia , Grafite/química , Humanos , Canais Iônicos/metabolismo , Potenciais da Membrana/efeitos dos fármacos , Proteínas de Membrana/metabolismo , Microscopia Eletrônica de Varredura , Oxirredução , Canais de Potássio de Abertura Dependente da Tensão da Membrana/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores de Superfície Celular/metabolismo , Espectroscopia de Infravermelho com Transformada de Fourier
4.
Medicina (Kaunas) ; 57(1)2020 Dec 31.
Artigo em Inglês | MEDLINE | ID: mdl-33396344

RESUMO

Revision hip arthroplasty procedures have been extensively discussed in the literature. At the same time, discussions of the management of acetabular component protrusion into the pelvic cavity, and, more specifically, the subperitoneal space, necessitating an additional abdominal approach for the revision arthroplasty, have only been published as case reports and descriptions of transperitoneal approaches have been even rarer. This paper presents the case of a 63-year-old female patient in whom a peritoneal approach was necessary to access a migrated acetabular component. The outcome of the treatment, which represented a complex orthopedic and general surgical problem, was good. We believe that the complexity of revision hip arthroplasty in patients with protrusion of the acetabular component together with the head and proximal part of the stem of the implant into subperitoneal space calls for a careful re-analysis of the category of Type III bony acetabulum defects according to Paprosky, where the recognition of two subtypes would facilitate analysis of such cases.


Assuntos
Artroplastia de Quadril , Prótese de Quadril , Acetábulo/diagnóstico por imagem , Acetábulo/cirurgia , Feminino , Seguimentos , Humanos , Pessoa de Meia-Idade , Falha de Prótese , Reoperação , Estudos Retrospectivos , Resultado do Tratamento
5.
BMC Mol Biol ; 20(1): 2, 2019 01 03.
Artigo em Inglês | MEDLINE | ID: mdl-30602369

RESUMO

BACKGROUND: Recently different forms of nanographene were proposed as the material with high anticancer potential. However, the mechanism of the suppressive activity of the graphene on cancer development remains unclear. We examined the effect of oxygenated, reduced and pristine graphene on the gene expression in glioblastoma U87 cell line. RESULTS: Conducting microarrays and RT-qPCR analysis we explored that graphene oxide (rather than reduced graphene oxide and pristine graphene) down-regulates the mRNA expression of mitochondrial oxidative phosphorylation (OXPHOS) nuclear genes of complexes I, III, IV and V. The presented results provide first evidence for the hypothesis that the suppressed growth of GBM can be the consequence of down-regulation of OXPHOS protein expression and decreased ATP level. CONCLUSIONS: We suggest that changes in the expression of OXPHOS genes identified in our study may mediate the anti-proliferative and anti-migratory effects of graphene oxide in glioblastoma cells. However, further investigations with different cell lines, regarding expression, regulation and activity of OXPHOS genes identified in our study is necessary to elucidate the mechanism mediating the anti-proliferative and anti-migratory effects of graphene oxide in glioblastoma cells.


Assuntos
Antineoplásicos/farmacologia , Expressão Gênica/efeitos dos fármacos , Glioblastoma/genética , Grafite/farmacologia , Nanopartículas , Linhagem Celular Tumoral , Regulação para Baixo , Humanos , Proteínas de Transporte da Membrana Mitocondrial/genética , Fosforilação Oxidativa/efeitos dos fármacos , RNA Mensageiro/metabolismo
6.
BMC Microbiol ; 19(1): 92, 2019 05 10.
Artigo em Inglês | MEDLINE | ID: mdl-31077130

RESUMO

BACKGROUND: Cathepsins are a group of endosomal proteases present in many cells including dendritic cells (DCs). The activity of cathepsins is regulated by their endogenous inhibitors - cystatins. Cathepsins are crucial to antigen processing during viral and bacterial infections, and as such are a prerequisite to antigen presentation in the context of major histocompatibility complex class I and II molecules. Due to the involvement of DCs in both innate and adaptive immune responses, and the quest to understand the impact of poxvirus infection on host cells, we investigated the influence of ectromelia virus (ECTV) infection on cathepsin and cystatin levels in murine conventional DCs (cDCs). ECTV is a poxvirus that has evolved many mechanisms to avoid host immune response and is able to replicate productively in DCs. RESULTS: Our results showed that ECTV-infection of JAWS II DCs and primary murine GM-CSF-derived bone marrow cells down-regulated both mRNA and protein of cathepsin B, L and S, and cystatin B and C, particularly during the later stages of infection. Moreover, the activity of cathepsin B, L and S was confirmed to be diminished especially at later stages of infection in JAWS II cells. Consequently, ECTV-infected DCs had diminished ability to endocytose and process a soluble antigen. Close examination of cellular protein distribution showed that beginning from early stages of infection, the remnants of cathepsin L and cystatin B co-localized and partially co-localized with viral replication centers (viral factories), respectively. Moreover, viral yield increased in cDCs treated with siRNA against cathepsin B, L or S and subsequently infected with ECTV. CONCLUSIONS: Taken together, our results indicate that infection of cDCs with ECTV suppresses cathepsins and cystatins, and alters their cellular distribution which impairs the cDC function. We propose this as an additional viral strategy to escape immune responses, enabling the virus to replicate effectively in infected cells.


Assuntos
Catepsinas/genética , Cistatinas/genética , Células Dendríticas/virologia , Vírus da Ectromelia/fisiologia , Animais , Células Dendríticas/imunologia , Regulação para Baixo , Endossomos/imunologia , Endossomos/virologia , Técnicas de Silenciamento de Genes , Masculino , Camundongos , Camundongos Endogâmicos C57BL , RNA Interferente Pequeno , Replicação Viral
7.
Int J Mol Sci ; 20(18)2019 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-31547375

RESUMO

Colorectal cancer ranks 3rd in terms of cancer incidence. Growth and development of colon cancer cells may be affected by juice and extracts from Saposhnikovia divaricata root. The objective of the research was to analyze the effect of S. divaricata juice and extracts on the viability, membrane integrity and types of cell death of Caco-2 cells. Juice and extracts were analyzed using Ultra-High Performance Liquid Chromatography-Mass Spectrometry (UHPLC-MS) and in respect of the presence of antioxidants, total carbohydrates, protein, fat and polyphenols. The contents of cimifugin ß-D-glucopyranoside, cimifugin, 4'-O-glucopyranosyl-5-O-methylvisamminol, imperatorin and protein were the highest in juice. 50% Hydroethanolic extract had the greatest antioxidant potential, concentration of polyphenols and fat. Water extract was characterized by the highest content of glutathione. Juice and 75% hydroethanolic extract contained the most carbohydrates. After the application of juice, 50% extract and the juice fraction containing the molecules with molecular weights >50 kDa, a decrease of the cell viability was noted. Juice and this extract exhibited the protective properties in relation to the cell membranes and they induced apoptosis. The knowledge of further mechanisms of anticancer activity of the examined products will allow to consider their use as part of combination therapy.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Apiaceae/química , Neoplasias do Colo/tratamento farmacológico , Extratos Vegetais/farmacologia , Animais , Antineoplásicos Fitogênicos/química , Apoptose/efeitos dos fármacos , Células CACO-2 , Humanos , Extratos Vegetais/química , Raízes de Plantas/química
8.
Int J Mol Sci ; 20(3)2019 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-30717385

RESUMO

Due to the development of nanotechnologies, graphene and graphene-based nanomaterials have attracted immense scientific interest owing to their extraordinary properties. Graphene can be used in many fields, including biomedicine. To date, little is known about the impact graphene may have on human health in the case of intentional exposure. The present study was carried out on U87 glioma cells and non-cancer HS-5 cell lines as in vitro model and U87 tumors cultured on chicken embryo chorioallantoic membrane as in vivo model, on which the effects of pristine graphene platelets (GPs) were evaluated. The investigation consisted of structural analysis of GPs using transmission electron microscopy, Fourier transmission infrared measurements, zeta potential measurements, evaluation of cell morphology, assessment of cell viability, investigation of reactive oxygen species production, and investigation of mitochondrial membrane potential. The toxicity of U87 glioma tumors was evaluated by calculating the weight and volume of tumors and performing analyses of the ultrastructure, histology, and protein expression. The in vitro results indicate that GPs have dose-dependent cytotoxicity via ROS overproduction and depletion of the mitochondrial membrane potential. The mass and volume of tumors were reduced in vivo after injection of GPs. Additionally, the level of apoptotic and necrotic markers increased in GPs-treated tumors.


Assuntos
Fulerenos/farmacologia , Grafite/farmacologia , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Apoptose/efeitos dos fármacos , Biomarcadores , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Fulerenos/química , Glioblastoma/metabolismo , Glioblastoma/patologia , Glioblastoma/ultraestrutura , Grafite/química , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Espectroscopia de Infravermelho com Transformada de Fourier
9.
Molecules ; 24(8)2019 Apr 19.
Artigo em Inglês | MEDLINE | ID: mdl-31010146

RESUMO

Our previous studies have shown that diamond nanoparticles (NDs) exhibited antiangiogenic and proapoptotic properties in vitro in glioblastoma multiforme (GBM) cells and in tumors in vivo. Moreover, NDs inhibited adhesion, leading to the suppression of migration and invasion of GBM. In the present study, we hypothesized that the NDs might also inhibit proliferation and cell cycle in glioma cells. Experiments were performed in vitro with the U87 and U118 lines of GBM cells, and for comparison, the Hs5 line of stromal cells (normal cells) after 24 h and 72 h of treatment. The analyses included cell morphology, cell death, viability, and cell cycle analysis, double timing assay, and gene expression (Rb, E2F1, CycA, CycB, CycD, CycE, PTEN, Ki-67). After 72 h of ND treatment, the expression level of Rb, CycD, and CycE in the U118 cells, and E2F1, CycD, and CycE in the U87 cells were significantly lower in comparison to those in the control group. We observed that decreased expression of cyclins inhibited the G1/S phase transition, arresting the cell cycle in the G0/G1 phase in glioma cells. The NDs did not affect the cell cycle as well as PTEN and Ki-67 expression in normal cells (Hs5), although it can be assumed that the NDs reduced proliferation and altered the cell cycle in fast dividing cells.


Assuntos
Diamante/química , Diamante/farmacologia , Glioblastoma/metabolismo , Glioma/metabolismo , Nanopartículas/química , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Ciclina B/metabolismo , Ciclina D/metabolismo , Ciclina E/metabolismo , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos
10.
Parasitol Res ; 117(11): 3519-3525, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30112674

RESUMO

Free-living amoebae belonging to Acanthamoeba genus are widely distributed protozoans which are able to cause infection in humans and other animals such as keratitis and encephalitis. Acanthamoeba keratitis is a vision-threatening corneal infection with currently no available fully effective treatment. Moreover, the available therapeutic options are insufficient and are very toxic to the eye. Therefore, there is an urgent need for the development of more effective anti-amoebic agents. Nanotechnology approaches have been recently reported to be useful for the elucidation antimicrobial, antiviral, antifungal and antiprotozoal activities and thus, they could be a good approach for the development of anti-Acanthamoeba agents. Therefore, this study was aimed to explore the activity and cytotoxicity of tannic acid-modified silver nanoparticles, pure silver nanoparticles and pure gold nanoparticles against clinical strains of Acanthamoeba spp. The obtained results showed a significant anti-amoebic effect of the tannic acid-modified silver nanoparticles which also presented low cytotoxicity. Moreover, tannic acid-modified silver nanoparticles were well absorbed by the trophozoites and did not induce encystation. On the other hand, pure silver nanoparticles were only slightly active against the trophozoite stage and pure gold nanoparticles did not show any activity. In conclusion and based on the observed results, silver nanoparticle conjugation with tannic acid may be considered as potential agent against Acanthamoeba spp.


Assuntos
Ceratite por Acanthamoeba/tratamento farmacológico , Acanthamoeba/efeitos dos fármacos , Amebíase/tratamento farmacológico , Antiprotozoários/uso terapêutico , Infecções Protozoárias do Sistema Nervoso Central/tratamento farmacológico , Encefalite Infecciosa/tratamento farmacológico , Nanopartículas Metálicas/uso terapêutico , Taninos/uso terapêutico , Ceratite por Acanthamoeba/parasitologia , Amebíase/parasitologia , Amoeba/efeitos dos fármacos , Animais , Infecções Protozoárias do Sistema Nervoso Central/parasitologia , Ouro/farmacologia , Humanos , Encefalite Infecciosa/parasitologia , Prata/farmacologia , Trofozoítos/efeitos dos fármacos
11.
Int J Mol Sci ; 19(4)2018 Apr 03.
Artigo em Inglês | MEDLINE | ID: mdl-29614018

RESUMO

Colorectal cancer is the third most widely diagnosed cancer. Extracts from snails may modulate growth and development of colorectal cancer cells. The objective of this study was to determine the chemical composition of tissues derived from Helix aspersa Müller and red-ox properties of tissue extracts. Then, the influence of extracts and their fractions of different molecular weights on viability of Caco-2 cells was examined. Tissue lyophilisates contained antioxidants that could be important in the prevention of colorectal cancer. Moreover, we confirmed the presence of a wide array of compounds that might be used in treatment of this disease. The decrease of cell viability after the application of extracts from lyophilized mucus and foot tissues was affirmed. The effect of extract from mucus could be related to the content of some proteins and peptides, proper essential amino acids (EAA)/non-essential amino acids (NEAA) ratio, Met restriction and the presence of Cu, Ca, Zn, Se. The influence of the extract from foot tissues could be assigned additionally to the presence of eicosapentaenoic, α-linolenic, linoleic and γ-linolenic acids. The opposite effect was demonstrated by extract from lyophilized shells which increased cell viability. Further studies are needed to know whether dietary supplying of H. aspersa Müller tissues can be used as an approach in colorectal cancer management.


Assuntos
Antineoplásicos/toxicidade , Caramujos/química , Extratos de Tecidos/toxicidade , Animais , Células CACO-2 , Sobrevivência Celular/efeitos dos fármacos , Humanos
12.
Int J Mol Sci ; 19(12)2018 Dec 07.
Artigo em Inglês | MEDLINE | ID: mdl-30544611

RESUMO

Graphene (GN) and its derivatives (rGOs) show anticancer properties in glioblastoma multiforme (GBM) cells in vitro and in tumors in vivo. We compared the anti-tumor effects of rGOs with different oxygen contents with those of GN, and determined the characteristics of rGOs useful in anti-glioblastoma therapy using the U87 glioblastoma line. GN/ExF, rGO/Term, rGO/ATS, and rGO/TUD were structurally analysed via transmission electron microscopy, Raman spectroscopy, FTIR, and AFM. Zeta potential, oxygen content, and electrical resistance were determined. We analyzed the viability, metabolic activity, apoptosis, mitochondrial membrane potential, and cell cycle. Caspase- and mitochondrial-dependent apoptotic pathways were investigated by analyzing gene expression. rGO/TUD induced the greatest decrease in the metabolic activity of U87 cells. rGO/Term induced the highest level of apoptosis compared with that induced by GN/ExF. rGO/ATS induced a greater decrease in mitochondrial membrane potential than GN/ExF. No significant changes were observed in the cytometric study of the cell cycle. The effectiveness of these graphene derivatives was related to the presence of oxygen-containing functional groups and electron clouds. Their cytotoxicity mechanism may involve electron clouds, which are smaller in rGOs, decreasing their cytotoxic effect. Overall, cytotoxic activity involved depolarization of the mitochondrial membrane potential and the induction of apoptosis in U87 glioblastoma cells.


Assuntos
Antineoplásicos/farmacologia , Grafite/química , Óxidos/farmacologia , Antineoplásicos/química , Apoptose/efeitos dos fármacos , Ciclo Celular , Linhagem Celular Tumoral , Glioblastoma/patologia , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Óxidos/química , Espécies Reativas de Oxigênio/metabolismo
13.
Int J Mol Sci ; 16(10): 25214-33, 2015 Oct 23.
Artigo em Inglês | MEDLINE | ID: mdl-26512645

RESUMO

Our previous studies revealed that graphene had anticancer properties in experiments in vitro with glioblastoma multiforme (GBM) cells and in tumors cultured in vivo. We hypothesized that the addition of arginine or proline to graphene solutions might counteract graphene agglomeration and increase the activity of graphene. Experiments were performed in vitro with GBM U87 cells and in vivo with GBM tumors cultured on chicken embryo chorioallantoic membranes. The measurements included cell morphology, mortality, viability, tumor morphology, histology, and gene expression. The cells and tumors were treated with reduced graphene oxide (rGO) and rGO functionalized with arginine (rGO + Arg) or proline (rGO + Pro). The results confirmed the anticancer effect of graphene on GBM cells and tumor tissue. After functionalization with amino acids, nanoparticles were distributed more specifically, and the flakes of graphene were less agglomerated. The molecule of rGO + Arg did not increase the expression of TP53 in comparison to rGO, but did not increase the expression of MDM2 or the MDM2/TP53 ratio in the tumor, suggesting that arginine may block MDM2 expression. The expression of NQO1, known to be a strong protector of p53 protein in tumor tissue, was greatly increased. The results indicate that the complex of rGO + Arg has potential in GBM therapy.


Assuntos
Antineoplásicos/farmacologia , Glioblastoma/metabolismo , Grafite/farmacologia , Animais , Antineoplásicos/química , Arginina/química , Linhagem Celular Tumoral , Proliferação de Células , Sobrevivência Celular , Embrião de Galinha , Grafite/química , NAD(P)H Desidrogenase (Quinona)/genética , NAD(P)H Desidrogenase (Quinona)/metabolismo , Neurônios/efeitos dos fármacos , Óxidos/química , Proteínas Proto-Oncogênicas c-mdm2/genética , Proteínas Proto-Oncogênicas c-mdm2/metabolismo , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo
14.
Arch Anim Nutr ; 69(1): 57-68, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25530495

RESUMO

It has been considered that concentrations of certain amino acids in the egg are not sufficient to fully support embryonic development of modern broilers. In this study we evaluated embryo growth and development with particular emphasis on one of the major components of connective tissue, collagen. Experiments were performed on Ross 308 chicken embryos from 160 fertilised eggs. Experimental solutions of silver nanoparticles (Ag), hydroxyproline solution (Hyp) and a complex of silver nanoparticles with hydroxyproline (AgHyp) were injected into albumen, and embryos were incubated until day 20. An assessment of the mass of embryo and selected organs was carried out followed by measurements of the expression of the key signalling factors' fibroblast growth factor-2 (FGF-2) and vascular endothelial growth factor-A (VEGF-A). Finally, an evaluation of collagen microstructure using scanning electron microscopy was performed. Our results clearly indicate that Hyp, Ag and AgHyp administered in ovo to chicken embryos did not harm embryos. Comparing to the control group, Hyp, Ag and the AgHyp complex significantly upregulated expression of the FGF-2 at the mRNA and protein levels. Moreover, Hyp, Ag and, in particular, the complex of AgHyp significantly increased blood vessel size, cartilage collagen fibre lattice size and bundle thickness. The general conclusion from this study is that AgHyp treatment may help to build a stronger and longer lasting form of collagen fibres.


Assuntos
Embrião de Galinha/efeitos dos fármacos , Hidroxiprolina/farmacologia , Nanopartículas Metálicas/química , Osteocondrodisplasias/metabolismo , Prata/farmacologia , Animais , Embrião de Galinha/irrigação sanguínea , Embrião de Galinha/metabolismo , Vasos Coronários/embriologia , Vasos Coronários/ultraestrutura , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Hidroxiprolina/administração & dosagem , Microscopia Eletrônica de Varredura , Prata/química
15.
ACS Appl Mater Interfaces ; 16(25): 32128-32146, 2024 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-38872576

RESUMO

Atopic dermatitis (AD) is a chronic inflammatory skin disease with a complex etiology that lacks effective treatment. The therapeutic goals include alleviating symptoms, such as moisturizing and applying antibacterial and anti-inflammatory medications. Hence, there is an urgent need to develop a patch that effectively alleviates most of the AD symptoms. In this study, we employed a "green" cross-linking approach of poly(vinyl alcohol) (PVA) using glycerol, and we combined it with polyacrylonitrile (PAN) to fabricate core-shell (CS) nanofibers through electrospinning. Our designed structure offers multiple benefits as the core ensures controlled drug release and increases the strength of the patch, while the shell provides skin moisturization and exudate absorption. The efficient PVA cross-linking method facilitates the inclusion of sensitive molecules such as fermented oils. In vitro studies demonstrate the patches' exceptional biocompatibility and efficacy in minimizing cell ingrowth into the CS structure containing argan oil, a property highly desirable for easy removal of the patch. Histological examinations conducted on an ex vivo model showed the nonirritant properties of developed patches. Furthermore, the eradication of Staphylococcus aureus bacteria confirms the potential use of CS nanofibers loaded with argan oil or norfloxacin, separately, as an antibacterial patch for infected AD wounds. In vivo patch application studies on patients, including one with AD, demonstrated ideal patches' moisturizing effect. This innovative approach shows significant promise in enhancing life quality for AD sufferers by improving skin hydration and avoiding infections.


Assuntos
Antibacterianos , Dermatite Atópica , Staphylococcus aureus , Dermatite Atópica/tratamento farmacológico , Dermatite Atópica/patologia , Antibacterianos/química , Antibacterianos/farmacologia , Humanos , Staphylococcus aureus/efeitos dos fármacos , Nanofibras/química , Adesivo Transdérmico , Adesivos/química , Adesivos/farmacologia , Nanoestruturas/química , Animais , Pele/efeitos dos fármacos , Pele/patologia
16.
Biomed Pharmacother ; 170: 116090, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-38169187

RESUMO

PURPOSE: The aim of the study was to evaluate the effect of silver nanoparticles hydrocolloids (AgNPs) on human corneal epithelial cells. Epithelial cells form the outermost and the most vulnerable to environmental stimuli layer of the cornea in the eye. Mechanical stress, UV radiation, and pathogens such as bacteria, viruses, and parasites challenge the fragile homeostasis of the eye. To help combat stress, infection, and inflammation wide portfolio of interventions is available. One of the oldest treatments is colloidal silver. Silver nanoparticle suspension in water is known for its anti-bacterial anti-viral and antiprotozoal action. However, AgNPs interact also with host cells, and the character of the interplay between corneal cells and silver seeks investigation. METHODS: The human epithelial corneal cell line (HCE-2) was cultured in vitro, treated with AgNPs, and subjected to UV. The cell's viability, migration, calcium concentration, and expression/protein level of selected proteins were investigated by appropriate methods including cytotoxicity tests, "wound healing" assay, Fluo8/Fura2 AM staining, qRT-PCR, and western blot. RESULTS: Incubation of human corneal cells (HCE-2) with AgNP did not affect cells viability but limited cells migration and resulted in altered calcium homeostasis, decreased the presence of ATP-activated P2X7, P2Y2 receptors, and enhanced the expression of PACAP. Furthermore, AgNPs pretreatment helped restrain some of the deleterious effects of UV irradiation. Interestingly, AgNPs had no impact on the protein level of ACE2, which is important in light of potential SARS-CoV-2 entrance through the cornea. CONCLUSIONS: Silver nanoparticles are safe for corneal epithelial cells in vitro.


Assuntos
Nanopartículas Metálicas , Prata , Humanos , Prata/metabolismo , Cálcio/metabolismo , Nanopartículas Metálicas/toxicidade , Receptores Purinérgicos P2Y2/metabolismo , Córnea , Células Epiteliais
17.
Curr Microbiol ; 67(6): 696-702, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23884593

RESUMO

Gene-nominated cj0183 was identified in Campylobacter jejuni NCTC 11168 and in two human isolates 81116 and 81-176. It encodes a protein which shows partial homology to TlyC of Brachyspira hyodysenteriae. The aim of this work was to determine the mechanisms of gene regulation by cloning DNA fragments lying upstream of the cj0183 gene. The ß-galactosidase activity determined for the strain harboring the plasmid with the fragment upstream of cj0183 indicated the presence of a promoter in this DNA region. Mutations in cj0183 -10 region, -16 region, and -35 region resulted in changes in gene transcription.


Assuntos
Campylobacter jejuni/genética , Análise Mutacional de DNA , Regiões Promotoras Genéticas , Fusão Gênica Artificial , Sequência de Bases , Expressão Gênica , Genes Reporter , Dados de Sequência Molecular , beta-Galactosidase/análise , beta-Galactosidase/genética
18.
Int J Mol Sci ; 14(11): 23033-44, 2013 Nov 20.
Artigo em Inglês | MEDLINE | ID: mdl-24264045

RESUMO

It has been demonstrated that the content of certain amino acids in eggs is not sufficient to fully support embryonic development. One possibility to supply the embryo with extra nutrients and energy is in ovo administration of nutrients. Nanoparticles of diamond are highly biocompatible non-toxic carbonic structures, and we hypothesized that bio-complexes of diamond nanoparticles with L-glutamine may affect molecular responses in breast muscle. The objective of the investigation was to evaluate the effect of diamond nanoparticle (ND) and L-glutamine (Gln) on expression of growth and differentiation factors of chicken embryo pectoral muscles. ND, Gln, and Gln/ND solutions (50 mg/L) were injected into fertilized broiler chicken eggs at the beginning of embryogenesis. Muscle tissue was dissected at day 20 of incubation and analysed for gene expression of FGF2, VEGF-A, and MyoD1. ND and especially Gln/ND up-regulated expression of genes related to muscle cell proliferation (FGF2) and differentiation (MyoD1). Furthermore, the ratio between FGF2 and MyoD1 was highest in the Gln/ND group. At the end of embryogenesis, Gln/ND enhanced both proliferation and differentiation of pectoral muscle cells and differentiation dominated over proliferation. These preliminary results suggest that the bio-complex of glutamine and diamond nanoparticles may accelerate growth and maturation of muscle cells.


Assuntos
Diamante/administração & dosagem , Desenvolvimento Embrionário , Nanopartículas/administração & dosagem , Músculos Peitorais/efeitos dos fármacos , Animais , Embrião de Galinha/efeitos dos fármacos , Embrião de Galinha/crescimento & desenvolvimento , Galinhas/crescimento & desenvolvimento , Glutamina/administração & dosagem , Avaliação Nutricional , Músculos Peitorais/crescimento & desenvolvimento
19.
Materials (Basel) ; 16(16)2023 Aug 12.
Artigo em Inglês | MEDLINE | ID: mdl-37629892

RESUMO

Diamond nanoparticles, also known as nanodiamonds (NDs), exhibit remarkable, awe-inspiring properties that make them suitable for various applications in the field of skin care products. However, a comprehensive assessment of their compatibility with human skin, according to the irritation criteria established by the Organization for Economic Cooperation and Development (OECD), has not yet been conducted. The purpose of this study was to evaluate if diamond nanoparticles at a concentration of 25 µg/mL, incubated with reconstituted human epidermis (EpiDermTM) for 18 h, conform to the OECD TG439 standard used to classify chemical irritants. For this purpose, a cell viability test (MTT assay), histological assessment, and analysis of pro-inflammatory cytokine expression were performed. The results indicated that NDs had no toxic effect at the tested concentration. They also did not adversely affect tissue structure and did not lead to a simultaneous increase in protein and mRNA expression of the analyzed cytokines. These results confirm the safety and biocompatibility of NDs for application in skincare products, thereby creating a wide range of possibilities to exert an impact on the advancement of contemporary cosmetology in the future.

20.
Nanotoxicology ; 17(4): 310-337, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-37262345

RESUMO

Reports on the cytotoxicity of diamond nanoparticles (ND) are ambiguous and depend on the physicochemical properties of the material and the tested cell lines. Thus, the aim of this research was to evaluate the influence of thirteen types of diamond nanoparticles, differing in production method, size, and surface functional groups, on their cytotoxicity against four tumor cell lines (T98G, U-118 MG, MCF-7, and Hep G2) and one non-tumor cell line (HFF-1). In order to understand the dependence of diamond nanoparticles on physicochemical properties, the following parameters were analyzed: viability, cell membrane damage, morphology, and the level of intracellular general ROS and mitochondrial superoxide. The performed analyses revealed that all diamond nanoparticles showed no toxicity to MCF-7, Hep G2, and HFF-1 cells. In contrast, the same nanomaterials were moderately toxic for the glioblastoma T98G and U-118 MG cell lines. In general, the effect of the production method did not influence ND toxicity. Some changes in cell response after treatment with modified nanomaterials were observed, with the presence of carboxyl groups having a more detrimental effect than the presence of other functional groups. Although nanoparticles of different sizes caused similar toxicity, nanomaterials with bigger particles caused a more pronounced effect.


Assuntos
Neoplasias da Mama , Carcinoma Hepatocelular , Glioblastoma , Neoplasias Hepáticas , Nanopartículas , Humanos , Feminino , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patologia , Glioblastoma/patologia , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patologia , Nanopartículas/toxicidade , Nanopartículas/química , Linhagem Celular Tumoral , Sobrevivência Celular
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