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1.
J Reconstr Microsurg ; 39(4): 311-319, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-35817403

RESUMO

BACKGROUND: Lymphedema is a chronic condition characterized by progressive edema with complicated treatment. Recently, new treatment strategies inducing lymphangiogenesis were proposed. The aim of our study was to examine the effect of vascular endothelial growth factor C (VEGF-C) and adipose-derived stem cells (ADSCs) on lymphatic regeneration and drainage re-establishment in vascularized lymph node transfer (VLNT) model using a pedicled vascularized lymph node (VLN) groin flap. METHODS: Female Lewis rats with groin VLN flaps were utilized as a lymphedema model. Group A served as the control. Group B received VEGF-C. Group C received both VEGF-C and ADSCs. Group D received ADSCs only. Lymphatic drainage re-establishment was evaluated by ultrasound-photoacoustic imaging (US-PAI) after indocyanine green (ICG) injection. RESULTS: The fastest regeneration of elevated flaps was observed in Groups B and C in all monitored periods. After the first month, ICG positivity was detected in 14.3% of animals in Group A, 71.43% of animals in Group B (odds ratio [OR] = 15; p = 0.048), and 83.33% in Group C (OR = 30; p = 0.027). On the contrary, the difference between control group and Group D (16.67%; p = 0.905) was statistically insignificant. Administration of VEGF-C, ADSC + VEGF-C, and ADSC led to full flap regeneration after 6 months. The control group had the lowest percentage of ICG positivity at all monitored time points. CONCLUSION: We found that the fastest regeneration occurred with the combination of the VLN flap and VEGF-C. The addition of ADSC had an insignificant effect in our study. Furthermore, we proved the feasibility of PAI as an assessment tool of the lymphatic drainage recovery in a VLNT model.


Assuntos
Linfedema , Fator C de Crescimento do Endotélio Vascular , Ratos , Feminino , Animais , Ratos Endogâmicos Lew , Linfonodos/irrigação sanguínea , Linfedema/cirurgia , Linfedema/etiologia , Verde de Indocianina , Regeneração , Células-Tronco
2.
Mol Biol Rep ; 46(4): 4483-4500, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31183678

RESUMO

One of the main aims of bone tissue engineering, regenerative medicine and cell therapy is development of an optimal artificial environment (scaffold) that can trigger a favorable response within the host tissue, it is well colonized by resident cells of organism and ideally, it can be in vitro pre-colonized by cells of interest to intensify the process of tissue regeneration. The aim of this study was to develop an effective tool for regenerative medicine, which combines the optimal bone-like scaffold and colonization technique suitable for cell application. Accordingly, this study includes material (physical, chemical and structural) and in vitro biological evaluation of scaffolds prior to in vivo study. Thus, porosity, permeability or elasticity of two types of bone-like scaffolds differing in the ratio of collagen type I and natural calcium phosphate nanoparticles (bCaP) were determined, then analyzes of scaffold interaction with mesenchymal stem cells (MSCs) were performed. Simultaneously, dynamic seeding using a perfusion bioreactor followed by static cultivation was compared with standard static cultivation for the whole period of cultivation. In summary, cell colonization ability was estimated by determination of cell distribution within the scaffold (number, depth and homogeneity), matrix metalloproteinase activity and gene expression analysis of signaling molecules and differentiation markers. Results showed, the used dynamic colonization technique together with the newly-developed collagen-based scaffold with high content of bCaP to be an effective combined tool for producing bone grafts for bone implantology and regenerative medicine.


Assuntos
Fosfatos de Cálcio/metabolismo , Células-Tronco Mesenquimais/metabolismo , Engenharia Tecidual/métodos , Animais , Osso e Ossos/química , Diferenciação Celular , Células Cultivadas , Colágeno/química , Feminino , Transplante de Células-Tronco Mesenquimais/métodos , Nanopartículas , Osteogênese/efeitos dos fármacos , Medicina Regenerativa , Suínos , Alicerces Teciduais/química
3.
J Mater Sci Mater Med ; 29(3): 20, 2018 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-29392427

RESUMO

Collagen composite scaffolds have been used for a number of studies in tissue engineering. The hydration of such highly porous and hydrophilic structures may influence mechanical behaviour and porosity due to swelling. The differences in physical properties following hydration would represent a significant limiting factor for the seeding, growth and differentiation of cells in vitro and the overall applicability of such hydrophilic materials in vivo. Scaffolds based on collagen matrix, poly(DL-lactide) nanofibers, calcium phosphate particles and sodium hyaluronate with 8 different material compositions were characterised in the dry and hydrated states using X-ray microcomputed tomography, compression tests, hydraulic permeability measurement, degradation tests and infrared spectrometry. Hydration, simulating the conditions of cell seeding and cultivation up to 48 h and 576 h, was found to exert a minor effect on the morphological parameters and permeability. Conversely, hydration had a major statistically significant effect on the mechanical behaviour of all the tested scaffolds. The elastic modulus and compressive strength of all the scaffolds decreased by ~95%. The quantitative results provided confirm the importance of analysing scaffolds in the hydrated rather than the dry state since the former more precisely simulates the real environment for which such materials are designed.


Assuntos
Colágeno/química , Dessecação , Alicerces Teciduais/química , Água/química , Materiais Biocompatíveis/química , Fosfatos de Cálcio/química , Força Compressiva , Módulo de Elasticidade , Ácido Hialurônico/química , Teste de Materiais , Fenômenos Mecânicos , Poliésteres/química , Porosidade , Engenharia Tecidual/métodos , Microtomografia por Raio-X
4.
Histochem Cell Biol ; 148(3): 273-288, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28432431

RESUMO

With the development of a wide range of new biomaterials for the sensing of different cell behaviour, it is important to consider whether the cells tested in vitro are in direct contact with the material or whether cell-biomaterial contact is mediated by an interfacial layer of proteins originating from the culture medium or from the cells themselves. Thus, this study describes the differences between the cell adhesion mediated by proteins originating from foetal bovine serum and without the presence of such proteins 2 h following cell seeding exemplarily with different cell types (an osteoblastic cell line, primary fibroblasts, and mesenchymal stem cells). Three of the examined cell types were found to react differently to differing conditions in terms of cell shape, area, and number. Nevertheless, the expression and localization of the various proteins involved in cell adhesion and signalling (CD44, vinculin, talin, actin, focal adhesion kinase, Rho-GTPases and extracellular signal-regulated kinases 1 and 2) were, in general, similar with respect to all the cell types tested, albeit varying according to the presence or absence of serum. Moreover, no classical focal adhesions were formed during cell adhesion without serum proteins, while different signalling pathways were involved in this process. The study systematically describes and discusses the cell adhesion of three different human cell types to a well-known substrate without the presence of external proteins and it is hoped that this knowledge will be subsequently applied in biomaterial applications in which the presence of external proteins is undesirable (e.g. for biosensing purposes).


Assuntos
Proteínas Sanguíneas/farmacologia , Adesão Celular/efeitos dos fármacos , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Proteínas Sanguíneas/química , Células Cultivadas , Humanos
5.
Small ; 12(18): 2499-509, 2016 05.
Artigo em Inglês | MEDLINE | ID: mdl-27000766

RESUMO

Two profoundly different carbon allotropes - nanocrystalline diamond and graphene - are of considerable interest from the viewpoint of a wide range of biomedical applications including implant coating, drug and gene delivery, cancer therapy, and biosensing. Osteoblast adhesion and proliferation on nanocrystalline diamond and graphene are compared under various conditions such as differences in wettability, topography, and the presence or absence of protein interlayers between cells and the substrate. The materials are characterized in detail by means of scanning electron microscopy, atomic force microscopy, photoelectron spectroscopy, Raman spectroscopy, and contact angle measurements. In vitro experiments have revealed a significantly higher degree of cell proliferation on graphene than on nanocrystalline diamond and a tissue culture polystyrene control material. Proliferation is promoted, in particular, by hydrophobic graphene with a large number of nanoscale wrinkles independent of the presence of a protein interlayer, i.e., substrate fouling is not a problematic issue in this respect. Nanowrinkled hydrophobic graphene, thus, exhibits superior characteristics for those biomedical applications where high cell proliferation is required under differing conditions.


Assuntos
Proliferação de Células/efeitos dos fármacos , Diamante/farmacologia , Grafite/farmacologia , Nanopartículas , Células-Tronco/efeitos dos fármacos , Células Cultivadas , Humanos , Microscopia de Força Atômica , Microscopia Eletrônica de Varredura , Espectroscopia Fotoeletrônica
6.
Heliyon ; 10(9): e30601, 2024 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-38742054

RESUMO

Stability and cytotoxicity of PEGylated Au NPs is crucial for biomedical application. In this study, we have focused on thermal stability of PEGylated Au NPs at 4 and 37 °C and after sterilization in autoclave. Gold nanoparticles were prepared by direct sputtering of gold into PEG and PEG-NH2. Transmission electron microscopy revealed that NPs exhibit a spherical shape with average dimensions 3.8 nm for both AuNP_PEG and AuNP_PEG-NH2. The single LSPR band at wavelength of 509 nm also confirmed presence of spherical Au NPs in both cases. Moreover, according to UV-Vis spectra, the Au NPs were overall stable during aging or thermal stressing and even after sterilization in autoclave. Based on gel electrophoresis results, the higher density of functionalizing ligands and the higher stability is assumed on AuNP_PEG-NH2. Changes in concentration of gold did not occur after thermal stress or with aging. pH values have to be adjusted to be suitable for bioapplications - original pH values are either too alkaline (AuNP_PEG-NH2, pH 10) or too acidic (AuNP_PEG, pH 5). Cytotoxicity was tested on human osteoblasts and fibroblasts. Overall, both Au NPs have shown good cytocompatibility either freshly prepared or even after Au NPs' sterilization in the autoclave. Prepared Au NP dispersions were also examined for their antiviral activity, however no significant effect was observed. We have synthesized highly stable, non-cytotoxic PEGylated Au NPs, which are ready for preclinical testing.

7.
Am J Hum Genet ; 85(2): 204-13, 2009 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-19664745

RESUMO

Through linkage analysis and candidate gene sequencing, we identified three unrelated families with the autosomal-dominant inheritance of early onset anemia, hypouricosuric hyperuricemia, progressive kidney failure, and mutations resulting either in the deletion (p.Leu16del) or the amino acid exchange (p.Leu16Arg) of a single leucine residue in the signal sequence of renin. Both mutations decrease signal sequence hydrophobicity and are predicted by bioinformatic analyses to damage targeting and cotranslational translocation of preprorenin into the endoplasmic reticulum (ER). Transfection and in vitro studies confirmed that both mutations affect ER translocation and processing of nascent preprorenin, resulting either in reduced (p.Leu16del) or abolished (p.Leu16Arg) prorenin and renin biosynthesis and secretion. Expression of renin and other components of the renin-angiotensin system was decreased accordingly in kidney biopsy specimens from affected individuals. Cells stably expressing the p.Leu16del protein showed activated ER stress, unfolded protein response, and reduced growth rate. It is likely that expression of the mutant proteins has a dominant toxic effect gradually reducing the viability of renin-expressing cells. This alters the intrarenal renin-angiotensin system and the juxtaglomerular apparatus functionality and leads to nephron dropout and progressive kidney failure. Our findings provide insight into the functionality of renin-angiotensin system and stress the importance of renin analysis in families and individuals with early onset hyperuricemia, anemia, and progressive kidney failure.


Assuntos
Anemia/genética , Genes Dominantes , Hiperuricemia/genética , Falência Renal Crônica/genética , Renina/genética , Adolescente , Adulto , Idade de Início , Anemia/metabolismo , Linhagem Celular , Criança , Pré-Escolar , Simulação por Computador , Feminino , Ligação Genética , Humanos , Hiperuricemia/metabolismo , Rim/citologia , Rim/ultraestrutura , Falência Renal Crônica/metabolismo , Masculino , Pessoa de Meia-Idade , Mutação , Linhagem , Renina/metabolismo , Análise de Sequência de DNA , Adulto Jovem
8.
Nanotoxicology ; 16(5): 610-628, 2022 06.
Artigo em Inglês | MEDLINE | ID: mdl-36170236

RESUMO

The study concerns the influence of graphene monolayer, as a 2 D platform, on cell viability, cytoskeleton, adhesions sites andmorphology of mitochondria of keratinocytes (HaCaT) under static conditions. Based on quantitative and immunofluorescent analysis, it could be stated that graphene substrate does not cause any damage to membrane or disruption of other monitored parameters. Spindle poles and cytokinesis bridges indicating proliferation of cells on this graphene substrate were detected. Moreover, the keratinocyte migration rate on the graphene substrate was comparable to control glass substrate when the created wound was completely closed after 38 hours. HaCaT morphology and viability were also assessed under dynamic conditions (lab on a chip - micro scale). For this purpose, microfluidic graphene system was designed and constructed. No differences as well as no anomalies were observed during cultivation of these cells on the graphene or glass substrates in relation to cultivation conditions: static (macro scale) and dynamic (micro scale). Only natural percentage of dead cells was determined using different methods, which proved that the graphene as the 2 D platform is cytocompatible with keratinocytes. The obtained results encourage the use of the designed lab on a chip system in toxicity testing of graphene also on other cells and further research on the use of graphene monolayers to produce bio-bandages for skin wounds in animal tests.


Assuntos
Grafite , Animais , Humanos , Grafite/toxicidade , Células HaCaT , Queratinócitos/metabolismo , Movimento Celular , Sobrevivência Celular , Proliferação de Células
9.
Int J Pharm ; 611: 121308, 2022 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-34800617

RESUMO

The utilization of nanoparticles for the intracellular delivery of theranostic agents faces one substantial limitation. Sequestration in intracellular vesicles prevents them from reaching the desired location in the cytoplasm or nucleus to deliver their cargo. We investigated whether three different cell-penetrating peptides (CPPs), namely, octa-arginine R8, polyhistidine KH27K and histidine-rich LAH4, could promote cytosolic and/or nuclear transfer of unique model nanoparticles-pseudovirions derived from murine polyomavirus. Two types of CPP-modified pseudovirions that carry the luciferase reporter gene were created: VirPorters-IN with CPPs genetically attached to the capsid interior and VirPorters-EX with CPPs noncovalently associated with the capsid exterior. We tested their transduction ability by luciferase assay and monitored their presence in subcellular fractions. Our results confirmed the overall effect of CPPs on the intracellular destination of the particles and suggested that KH27K has the potential to improve the cytosolic release of pseudovirions. None of the VirPorters caused endomembrane damage detectable by the Galectin-3 assay. Remarkably, a noncovalent modification was required to promote high transduction of the reporter gene and cytosolic delivery of pseudovirions mediated by LAH4. Together, CPPs in different arrangements have demonstrated their potential to improve pseudovirion invasion into cells, and these findings could be useful for the development of other nanoparticle-based delivery systems.


Assuntos
Peptídeos Penetradores de Células , Animais , Bioensaio , Cátions , Citosol , Histidina , Camundongos
10.
Stem Cells Int ; 2021: 6659244, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33727935

RESUMO

In vitro and in vivo analyses are closely connected, and the reciprocal relationship between the two comprises a key assumption with concern to the conducting of meaningful research. The primary purpose of in vitro analysis is to provide a solid background for in vivo and clinical study purposes. The fields of cell therapy, tissue engineering, and regenerative medicine depend upon the high quality and appropriate degree of the expansion of mesenchymal stromal cells (MSCs) under low-risk and well-defined conditions. Hence, it is necessary to determine suitable alternatives to fetal bovine serum (FBS-the laboratory gold standard) that comply with all the relevant clinical requirements and that provide the appropriate quantity of high-quality cells while preserving the required properties. Human serum (autologous and allogeneic) and blood platelet lysates and releasates are currently considered to offer promising and relatively well-accessible MSC cultivation alternatives. Our study compared the effect of heat-inactivated FBS on MSC metabolism as compared to its native form (both are used as the standard in laboratory practice) and to potential alternatives with concern to clinical application-human serum (allogeneic and autologous) or platelet releasate (PR-SRGF). The influence of the origin of the serum (fetal versus adult) was also determined. The results revealed the key impact of the heat inactivation of FBS on MSCs and the effectiveness of human sera and platelet releasates with respect to MSC behaviour (metabolic activity, proliferation, morphology, and cytokine production).

11.
Materials (Basel) ; 14(20)2021 Oct 14.
Artigo em Inglês | MEDLINE | ID: mdl-34683656

RESUMO

The paper examines the surface functionalization of a new type of Ti-graphite composite, a dental biomaterial prepared by vacuum low-temperature extrusion of hydrogenated-dehydrogenated titanium powder mixed with graphite flakes. Two experimental surfaces were prepared by laser micromachining applying different levels of incident energy of the fiber nanosecond laser working at 1064 nm wavelength. The surface integrity of the machined surfaces was evaluated, including surface roughness parameters measurement by contact profilometry and confocal laser scanning microscopy. The chemical and phase composition were comprehensively evaluated by scanning electron microscopy, energy-dispersive X-ray spectroscopy and X-ray diffraction analyses. Finally, the in vitro tests using human mesenchymal stem cells were conducted to compare the influence of the laser processing parameters used on the cell's cultivation and osteo-differentiation. The bioactivity results confirmed that the surface profile with positive kurtosis, platykurtic distribution curve and higher value of peaks spacing exhibited better bioactivity compared to the surface profile with negative kurtosis coefficient, leptokurtic distribution curve and lower peaks spacing.

12.
J Mater Chem B ; 9(36): 7386-7400, 2021 09 22.
Artigo em Inglês | MEDLINE | ID: mdl-34551046

RESUMO

Cerium oxide nanoparticles (CeNPs) possess multiple redox enzyme mimetic activities in scavenging reactive oxygen species (ROS) as a potential biomedicine. These enzymatic activities of CeNPs are closely related to their surface oxidation state. Here we have reported a synthetic method to modify CeNPs' surface oxidation state by changing the conformation of the poly(acrylic acid) (PAA) polymers adsorbed onto the CeNP surface. The synthesized PAA-CeNPs exhibited the same core size, morphology, crystal structure, and colloidal stability, with the only variation being their surface oxidation state (Ce3+ percentage). The modification mechanism can be attributed to the polymers chemisorbed onto the metal oxide surface forming a metal complexation structure. Such adsorption further modified CeNPs' surface oxidation state in a temperature-dependent manner. The series of PAA-CeNPs exhibited multiple redox enzyme mimetic activities (superoxide dismutase, catalase, peroxidase, and oxidase) directly related to their surface oxidation state. In vitro experiments showed no cytotoxic effect of these PAA-CeNPs on the osteoblastic cell line SAOS-2 at high loadings. Microscopic images confirmed the internalization of PAA-CeNPs in the cells. All tested PAA-CeNPs can reduce the basal and hydrogen peroxide-induced intracellular ROS level in the cells, indicating their effective intracellular ROS scavenging effect. However, we did not observe a positive correlation between the CeNP surface oxidation state and their capacities to reduce the intracellular ROS levels. We propose that CeNPs can maintain a dynamic state of Ce3+/Ce4+ during their catalytic activities, exhibiting a non-linear correlation between the CeNP surface oxidation state and their effect on regulating the intracellular ROS level.


Assuntos
Resinas Acrílicas/química , Cério/química , Nanopartículas Metálicas/química , Espécies Reativas de Oxigênio/metabolismo , Antioxidantes/química , Catálise , Linhagem Celular Tumoral , Humanos , Peróxido de Hidrogênio/farmacologia , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , Oxirredução , Tamanho da Partícula , Espécies Reativas de Oxigênio/química
13.
J Mater Chem B ; 9(40): 8530, 2021 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-34622917

RESUMO

Correction for 'Poly(acrylic acid)-mediated synthesis of cerium oxide nanoparticles with variable oxidation states and their effect on regulating the intracellular ROS level' by Xiaohui Ju et al., J. Mater. Chem. B, 2021, 9, 7386-7400, DOI: 10.1039/D1TB00706H.

14.
Materials (Basel) ; 14(3)2021 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-33573304

RESUMO

This study investigates the effect of graphene scaffold on morphology, viability, cytoskeleton, focal contacts, mitochondrial network morphology and activity in BALB/3T3 fibroblasts and provides new data on biocompatibility of the "graphene-family nanomaterials". We used graphene monolayer applied onto glass cover slide by electrochemical delamination method and regular glass cover slide, as a reference. The morphology of fibroblasts growing on graphene was unaltered, and the cell viability was 95% compared to control cells on non-coated glass slide. There was no significant difference in the cell size (spreading) between both groups studied. Graphene platform significantly increased BALB/3T3 cell mitochondrial activity (WST-8 test) compared to glass substrate. To demonstrate the variability in focal contacts pattern, the effect of graphene on vinculin was examined, which revealed a significant increase in focal contact size comparing to control-glass slide. There was no disruption in mitochondrial network morphology, which was branched and well connected in relation to the control group. Evaluation of the JC-1 red/green fluorescence intensity ratio revealed similar levels of mitochondrial membrane potential in cells growing on graphene-coated and uncoated slides. These results indicate that graphene monolayer scaffold is cytocompatible with connective tissue cells examined and could be beneficial for tissue engineering therapy.

15.
Clin Oral Implants Res ; 21(2): 174-81, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19709059

RESUMO

OBJECTIVES: Zirconia is a suitable biomaterial for use in medicine (stomatology, orthopaedics) due to its good biocompatibility and outstanding mechanical properties. This study compares the effect of (i) zirconia to the widely used titanium and (ii) zirconia with two different surface topographies (sandblasted and sandblasted/etched) on the adhesion, proliferation and differentiation of SAOS-2 osteoblasts. METHODS: SAOS-2 cells were cultured on either sandblasted or sandblasted/etched zirconia and compared with sandblasted/etched titanium. 2 and 24 h after plating, cell morphology was investigated by scanning electron microscope (SEM) and fluorescence imaging. At 24 and 48 h, cell number-relevant parameters were determined. Alkaline phosphatase (ALP) activity and mineral accumulation were measured at days 8, 11, 15 and day 22 of culture, respectively. RESULTS: SEM and fluorescence images revealed a faster spreading as well as higher number of adherent cells after 24 h incubation on zirconia compared with titanium. Also, the cellular metabolic activity after 24 h and the proliferation rate after 48 h is higher with zirconia compared with titanium. Zirconia had a more pronounced effect compared with titanium on the differentiation of SAOS-2 cells: ALP activity, an early differentiation marker increased earlier and mineralization, a late differentiation marker was increased. Only minor differences were found between zirconia with two different surface topographies; etched zirconia promoted slightly greater the differentiation of SAOS-2 cells. CONCLUSIONS: These data indicate that zirconia mediates a pronounced stronger effect on the adhesion, proliferation and differentiation compared with titanium; and that topographical differences of zirconia have minor effects on osteoblast biology.


Assuntos
Cerâmica/farmacologia , Osteoblastos/efeitos dos fármacos , Zircônio/farmacologia , Condicionamento Ácido do Dente , Abrasão Dental por Ar , Fosfatase Alcalina/metabolismo , Análise de Variância , Materiais Biocompatíveis/farmacologia , Adesão Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Microscopia Eletrônica de Varredura , Osteoblastos/citologia , Osteoblastos/enzimologia , Propriedades de Superfície , Titânio/farmacologia
16.
RSC Adv ; 10(65): 39373-39384, 2020 Oct 27.
Artigo em Inglês | MEDLINE | ID: mdl-35515371

RESUMO

One of the biggest challenges for the biomedical applications of cerium oxide nanoparticles (CeNPs) is to maintain their colloidal stability and catalytic activity as enzyme mimetics after nanoparticles enter the human cellular environment. This work examines the influences of CeNP surface properties on their colloidal stability and catalytic activity in cell culture media (CCM). Near-spherical CeNPs stabilized via different hydrophilic polymers were prepared through a wet-chemical precipitation method. CeNPs were stabilized via either electrostatic forces, steric forces, or a combination of both, generated by surface functionalization. CeNPs with electrostatic stabilization adsorb more proteins compared to CeNPs with only steric stabilization. The protein coverage further improves CeNPs colloidal stability in CCM. CeNPs with steric polymer stabilizations exhibited better resistance against agglomeration caused by the high ionic strength in CCM. These results suggest a strong correlation between CeNPs intrinsic surface properties and the extrinsic influences of the environment. The most stabilized sample in CCM is poly(acrylic acid) coated CeNPs (PAA-CeNPs), with a combination of both electrostatic and steric forces on the surface. It shows a hydrodynamic diameter of 15 nm while preserving 90% of its antioxidant activity in CCM. PAA-CeNPs are non-toxic to the osteoblastic cell line SAOS-2 and exhibit promising potential as a therapeutic alternative.

17.
Nanomaterials (Basel) ; 10(3)2020 Mar 22.
Artigo em Inglês | MEDLINE | ID: mdl-32235697

RESUMO

Ultra-small nanoparticles with sizes comparable to those of pores in the cellular membrane possess significant potential for application in the field of biomedicine. Silicon carbide ultra-small nanoparticles with varying surface termination were tested for the biological system represented by different human cells (using a human osteoblastic cell line as the reference system and a monocyte/macrophage cell line as immune cells). The three tested nanoparticle surface terminations resulted in the observation of different effects on cell metabolic activity. These effects were mostly noticeable in cases of monocytic cells, where each type of particle caused a completely different response ('as-prepared' particles, i.e., were highly cytotoxic, -OH terminated particles slightly increased the metabolic activity, while -NH2 terminated particles caused an almost doubled metabolic activity) after 24 h of incubation. Subsequently, the release of cytokines from such treated monocytes and their differentiation into activated cells was determined. The results revealed the potential modulation of immune cell behavior following stimulation with particular ultra-small nanoparticles, thus opening up new fields for novel silicon carbide nanoparticle biomedical applications.

18.
Int J Pharm ; 576: 119008, 2020 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-31901358

RESUMO

Viral nanoparticles represent potential natural versatile platforms for targeted gene and drug delivery. Improving the efficiency of gene transfer mediated by viral vectors could not only enhance their therapeutic potential, but also contribute to understanding the limitations in interactions of nanoparticles with cells and the development of new therapeutic approaches. In this study, four cell-penetrating peptides (CPPs), cationic octaarginine (R8), histidine-rich peptides (LAH4 and KH27K) and fusogenic peptide (FUSO), are investigated for their effect on infection by mouse polyomavirus (MPyV) or on transduction of reporter genes delivered by MPyV or related viral vectors. Peptides noncovalently associated with viral particles enhance gene transfer (with the exception of FUSO). Removal of cellular heparan sulfates by the heparinase does not significantly change the enhancing potential of CPPs. Instead, CPPs influences the physical state of viral particles: R8 slightly destabilizes the intact virus, KH27K induces its aggregation and LAH4 promotes disassembly and aggregation of the particles that massively and rapidly associate with cells. The findings indicate that peptides acting as transduction-enhancing agents of polyomavirus-based nanoparticles modulate their physical state, which can be an important prerequisite for sensitization of cells and determination of the further fate of viral particles inside cells.


Assuntos
Peptídeos Penetradores de Células/metabolismo , Vetores Genéticos , Polyomavirus/metabolismo , Transdução Genética , Vírion/metabolismo , Animais , Capsídeo/metabolismo , Capsídeo/ultraestrutura , Peptídeos Penetradores de Células/química , Células HEK293 , Humanos , Camundongos , Oligopeptídeos/química , Oligopeptídeos/metabolismo , Polyomavirus/genética , Polyomavirus/ultraestrutura , Vírion/genética , Vírion/ultraestrutura
19.
Sensors (Basel) ; 9(5): 3549-62, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-22412325

RESUMO

Microscopic chemical patterning of diamond surfaces by hydrogen and oxygen surface atoms is used for self-assembly of human osteoblastic cells into micro-arrays. The cell adhesion and assembly is further controlled by concentration of cells (2,500-10,000 cells/cm(2)) and fetal bovine serum (0-15%). The cells are characterized by fluorescence microscopy of actin fibers and nuclei. The serum protein adsorption is studied by atomic force microscopy (AFM). The cells are arranged selectively on O-terminated patterns into 30-200 µm wide arrays. Higher cell concentrations allow colonization of unfavorable H-terminated regions due to mutual cell communication. There is no cell selectivity without the proteins in the medium. Based on the AFM, the proteins are present on both H- and O-terminated surfaces. Pronounced differences in their thickness, surface roughness, morphology, and phase images indicate different conformation of the proteins and explain the cell selectivity.

20.
Materials (Basel) ; 12(17)2019 Aug 22.
Artigo em Inglês | MEDLINE | ID: mdl-31443361

RESUMO

Viral particles (VPs) have evolved so as to efficiently enter target cells and to deliver their genetic material. The current state of knowledge allows us to use VPs in the field of biomedicine as nanoparticles that are safe, easy to manipulate, inherently biocompatible, biodegradable, and capable of transporting various cargoes into specific cells. Despite the fact that these virus-based nanoparticles constitute the most common vectors used in clinical practice, the need remains for further improvement in this area. The aim of this review is to discuss the potential for enhancing the efficiency and versatility of VPs via their functionalization with cell-penetrating peptides (CPPs), short peptides that are able to translocate across cellular membranes and to transport various substances with them. The review provides and describes various examples of and means of exploitation of CPPs in order to enhance the delivery of VPs into permissive cells and/or to allow them to enter a broad range of cell types. Moreover, it is possible that CPPs are capable of changing the immunogenic properties of VPs, which could lead to an improvement in their clinical application. The review also discusses strategies aimed at the modification of VPs by CPPs so as to create a useful cargo delivery tool.

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