[Show abstract][Hide abstract] ABSTRACT: The pathophysiology of periapical lesions (PLs) is under control of pro-inflammatory and anti-inflammatory (mainly immunoregulatory) cytokines. We have recently established mesenchymal stem cells (MSCs) from PLs and showed their suppressive effects on the production of proinflammatory cytokines from PLs inflammatory cells (ICs). In this work we studied the production of interleukin (IL)-10, IL-27 and transforming growth factor (TGF)-β, by PL-ICs in direct or indirect contacts with PL-MSCs. PL-ICs, which were isolated from four different asymptomatic PLs, predominantly composed of lymphocytes, followed by neutrophil granulocytes, macrophages and plasma cells. PLMSCs, expressing typical MSC markers, were co-cultivated with PL-ICs at 1:10 ratio, either in direct contact or in a transwell-system, for 24 hours. The levels of cytokines in cell-culture supernatants were tested by ELISA. The results showed that PL-MSCs up-regulated the production of all three immunoregulatory cytokines by PL-ICs. PL-MSCs stimulated the production of IL-10 and IL-27 via soluble factors, whereas the up-regulation of TGF-β required direct cell-to-cell contacts. In conclusion, our results showed for the first time the involvement of PL-MSCs in restriction of inflammation in PLs by up-regulation of immunoregulatory cytokines.
[Show abstract][Hide abstract] ABSTRACT: Hashimoto thyroiditis (HT) is the most frequent thyroid autoimmune disease, while papillary thyroid cancer (PTC) is one of the most common endocrine malignancies. A few patients with HT also develop PTC. The aim of this study was to analyze cytokine profiles in patients with PTC accompanied with autoimmune HT in comparison with those in patients with PTC alone or HT alone and healthy subjects. Cytokine levels were determined in supernatants obtained from phytohemagglutinin (PHA)-stimulated whole blood cultures in vitro. The concentrations of selected cytokines: Th1-interferon gamma (IFN-γ); Th2-interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 6 (IL-6), interleukin 10 (IL-10) and interleukin 13 (IL-13); Th9-interleukin 9 (IL-9); and Th17-interleukin 17 (IL-17A) were measured using multiplex cytokine detection systems for human Th1/Th2/Th9/Th17/Th22. We found that PTC patients with HT produced significantly higher concentrations of IL-4, IL-6, IL-9, IL-13 and IFN-γ than PTC patients without HT. In conclusion, autoimmune HT affects the cytokine profile of patients with PTC by stimulating secretion of Th1/Th2/Th9 types of cytokines. Th1/Th2 cytokine ratios in PTC patients with associated autoimmune HT indicate a marked shift toward Th2 immunity.
Cancer Immunology and Immunotherapy 05/2015; 64(8). DOI:10.1007/s00262-015-1705-5 · 3.94 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Background & objectives:
Aluminum (Al) toxicity is closely linked to the pathogenesis of Alzheimer's disease (AD). This experimental study was aimed to investigate the active avoidance behaviour of rats after intrahippocampal injection of Al, and biochemical and immunohistochemical changes in three bilateral brain structures namely, forebrain cortex (FBCx), hippocampus and basal forebrain (BF).
Seven days after intra-hippocampal (CA1 sector) injection of AlCl3 into adult male Wistar rats they were subjected to two-way active avoidance (AA) tests over five consecutive days. Control rats were treated with 0.9% w/v saline. The animals were decapitated on the day 12 post-injection. The activities of acetylcholinesterase (AChE) and glucose-6-phosphate dehydrogenase (G6PDH) were measured in the FBCx, hippocampus and BF. Immunohistochemical staining was performed for transferrin receptors, amyloid β and tau protein.
The activities of both AChE and G6PDH were found to be decreased bilaterally in the FBCx, hippocampus and basal forebrain compared to those of control rats. The number of correct AA responses was reduced by AlCl3 treatment. G6PDH administered prior to AlCl3 resulted in a reversal of the effects of AlCl3 on both biochemical and behavioural parameters. Strong immunohistochemical staining of transferrin receptors was found bilaterally in the FBCx and the hippocampus in all three study groups. In addition, very strong amyloid β staining was detected bilaterally in all structures in AlCl3-treated rats but was moderate in G6PDH/AlCl3-treated rats. Strong tau staining was noted bilaterally in AlCl3-treated rats. In contrast, tau staining was only moderate in G6PDH/AlCl3-treated rats.
Interpretation & conclusions:
Our findings indicated that the G6PDH alleviated the signs of behavioural and biochemical effects of AlCl3-treatment suggesting its involvement in the pathogenesis of Al neurotoxicity and its potential therapeutic benefit. The present model could serve as a useful tool in AD investigations.
The Indian Journal of Medical Research 06/2014; 139(6):864-72. · 1.40 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Gold nanoparticles (GNPs) are claimed as outstanding biomedical tools for cancer diagnostics and photo-thermal therapy, but without enough evidence on their potentially adverse immunological effects. Using a model of human dendritic cells (DCs), we showed that 10 nm- and 50 nm-sized GNPs (GNP10 and GNP50, respectively) were internalized predominantly via dynamin-dependent mechanisms, and they both impaired LPS-induced maturation and allostimulatory capacity of DCs, although the effect of GNP10 was more prominent. However, GNP10 inhibited LPS-induced production of IL-12p70 by DCs, and potentiated their Th2 polarization capacity, while GNP50 promoted Th17 polarization. Such effects of GNP10 correlated with a stronger inhibition of LPS-induced changes in Ca2+ oscillations, their higher number per DC, and more frequent extra-endosomal localization, as judged by live-cell imaging, proton, and electron microscopy, respectively. Even when released from heat-killed necrotic HEp-2 cells, GNP10 inhibited the necrotic tumor cell-induced maturation and functions of DCs, potentiated their Th2/Th17 polarization capacity, and thus, impaired the DCs' capacity to induce T cell-mediated anti-tumor cytotoxicity in vitro. Therefore, GNP10 could potentially induce more adverse DC-mediated immunological effects, compared to GNP50.
PLoS ONE 05/2014; 9(5):e96584. DOI:10.1371/journal.pone.0096584 · 3.23 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Royal jelly (RJ) fatty acids have recently been shown to possess various pharmacological and biological activities. In this work, we studied the immunomodulatory effects of 10-hydroxy-trans-2-decenoic acid (10-HDA) and 3,10-dihydroxy-decanoic acid (3,10-DDA), isolated from RJ, using a model of phytohaemagglutinin-activated human peripheral blood mononuclear cells (PBMCs). We showed that higher concentrations (500 μM) of both fatty acids inhibited the proliferation of PBMCs, and the process was followed by a decrease in the production of interleukin-2 (IL-2). 10-HDA at the concentration of 500 μM inhibited the production of IL-1β and tumor necrosis factor-α by stimulated PBMCs, whereas the same dose of 3,10-DDA had no effect on the levels of these cytokines. Regarding T helper (Th) cytokine profile, higher concentration of 10-HDA, in contrast to the lower one (50 μM), inhibited both Th1 and Th2 response, whereas Th17 response was not significantly modulated, as judged by the levels of interferon-γ, IL-5 and IL-17A in culture supernatants, respectively. Lower concentration of 3,10-DDA stimulated Th1 and Th17 responses and inhibited IL-10 production, whereas the higher dose augmented the Th2 response. In conclusion, our results showed a significant, dose-dependent, immunomodulatory effect of RJ fatty acids in vitro, which was also associated with their structure.
European Food Research and Technology 05/2014; 238(5):881-887. DOI:10.1007/s00217-014-2154-7 · 1.56 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Immunoinflammatory mediated demyelination, the main pathological feature of multiple sclerosis (MS), is regularly accompanied by neurodegenerative processes, mostly in the form of axonal degeneration, which could be initiated by glutamate excitotoxicity. In the current study, the relationship between Th17 mediated inflammatory and excitotoxic events was investigated during an active phase of MS. Cerebrospinal fluid (CSF) of MS patients and control subjects was collected and IL-17A and glutamate levels were determined. IL-17A level was significantly higher in MS patients; whereas no statistically significant changes in glutamate concentrations were found. There was a direct correlation between IL-17A and glutamate levels; IL-17A levels were also associated with the neutrophil expansion in CSF and blood brain barrier disruption. However, IL-17A level and the number of neutrophils tended to fall with disease duration. The results suggest that Th17 cells might enhance and use glutamate excitotoxicity as an effector mechanism in the MS pathogenesis. Furthermore, Th17 immune response, as well as neutrophils, could be more important for MS onset rather than further disease development and progression, what could explain why some MS clinical trials, targeting Th17 cells in the later stage of the disease, failed to provide any clinical benefit. This article is protected by copyright. All rights reserved.
Scandinavian Journal of Immunology 01/2014; DOI:10.1111/sji.12147 · 1.74 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The epithelial cell adhesion molecule E-cadherin is important for maintenance normal tissue architecture and for cell-cell communication and immune cell migration. E-cadherin is also present in cholesteatomas. This study determined E-cadherin expression in acquired cholesteatomas and analyzed its expression according to cholesteatoma clinical and histological characteristics.
We investigated E-cadherin expression in 30 samples from operated patients with acquired middle ear cholesteatomas that were classified according to their clinical and histological characteristics. E-cadherin expression in cholesteatoma was determined immunohistochemically. A semi quantitative method was used to determine the index of expression of E-cadherin and t-tests, Mann-Whitney U tests and Spearman correlation analysis were used for statistical analysis.
We found significant expression of E-cadherin on CD1, CD3 total, CD4 (p < 0.05), high expression of E-cadherin on CD8 total and CD19/CD38 lymphocytes (p < 0.01) and very high expression of E-cadherin on mast cells and antigen-presenting cells, including Langerhans cells (p < 0.005). We graduated results as no statistically significant (p>0.05), statistically significant (0.05 > p > 0.01), highly statistically significant (0.01 > p > 0.005) and very highly statistically significant (p < 0.005).
E-cadherin expression was the same in the cholesteatoma matrix in all samples. There were no differences in expression according to the clinical and histological characteristics of the cholesteatomas.
[Show abstract][Hide abstract] ABSTRACT: Nickel-chromium (Ni-Cr) dental alloys have been widely used in prosthodontic practice, but there is a permanent concern about their biocompatibility due to the release of metal ions. This is especially important when Ni-Cr metal microparticles are incorporated into gingival tissue during prosthodontic procedures. Therefore, the aim of this study was to examine and compare the corrosion and cytotoxic properties of compact specimens and microparticles of Ni-Cr dental alloy.
Ni-Cr alloy, Remanium CSe bars (4 mm diameter), were made by the standard casting method and then cut into 0.5-mm-thick disks. Metal particles were obtained by scraping the bars using a diamond instrument for crown preparation. The microstructure was observed by an optical microscope. Quantitative determination and morphological and dimensional characterization of metal particles were carried out by a scanning electron microscope and Leica Application Suite software for image analysis. Corrosion was studied by conditioning the alloy specimens in the RPMI 1640 medium, containing 10% fetal calf serum in an incubator with 5% CO2 for 72 hours at 37°C. Inductively coupled plasma-optical emission spectrometry was used to assess metal ion release. The cytotoxity of conditioning medium (CM) was investigated on L929 cells using an MTT test. One-way ANOVA was used for statistical analysis.
After casting, the microstructure of the Remanium CSe compact specimen composed of Ni, Cr, Mo, Si, Fe, Al, and Co had a typical dendritic structure. Alloy microparticles had an irregular shape with a wide size range: from less than 1 μm to more than 100 μm. The release of metal ions, especially Ni and Mo from microparticles, was significantly higher, compared to the compact alloy specimen. The CM prepared from compact alloy was not cytotoxic at any tested dilutions, whereas CM from alloy microparticles showed dose-dependent cytotoxicity (90% CM and 45% CM versus control; p < 0.005).
Ni-Cr microparticles showed less corrosion resistance and lower biocompatibility than compact alloy. This could affect health on long-term exposure, especially in sensitized individuals.
Journal of Prosthodontics 10/2013; 23(3). DOI:10.1111/jopr.12100 · 1.07 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: OBJECTIVES:
ACST-2 is currently the largest trial ever conducted to compare carotid artery stenting (CAS) with carotid endarterectomy (CEA) in patients with severe asymptomatic carotid stenosis requiring revascularization.
Patients are entered into ACST-2 when revascularization is felt to be clearly indicated, when CEA and CAS are both possible, but where there is substantial uncertainty as to which is most appropriate. Trial surgeons and interventionalists are expected to use their usual techniques and CE-approved devices. We report baseline characteristics and blinded combined interim results for 30-day mortality and major morbidity for 986 patients in the ongoing trial up to September 2012.
A total of 986 patients (687 men, 299 women), mean age 68.7 years (SD ± 8.1) were randomized equally to CEA or CAS. Most (96%) had ipsilateral stenosis of 70-99% (median 80%) with contralateral stenoses of 50-99% in 30% and contralateral occlusion in 8%. Patients were on appropriate medical treatment. For 691 patients undergoing intervention with at least 1-month follow-up and Rankin scoring at 6 months for any stroke, the overall serious cardiovascular event rate of periprocedural (within 30 days) disabling stroke, fatal myocardial infarction, and death at 30 days was 1.0%.
Early ACST-2 results suggest contemporary carotid intervention for asymptomatic stenosis has a low risk of serious morbidity and mortality, on par with other recent trials. The trial continues to recruit, to monitor periprocedural events and all types of stroke, aiming to randomize up to 5,000 patients to determine any differential outcomes between interventions. Clinical trial: ISRCTN21144362.
European Journal of Vascular and Endovascular Surgery 09/2013; 46(5):510. DOI:10.1016/j.ejvs.2013.07.020 · 2.49 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The granulomatous foreign-body reaction (GFBR) is a special type of chronic inflammation characterised by infiltration of inflammatory cells (ICs). Using a model of subcutaneous implantation of polyvinyl sponges in rats, we studied the phenotypic and functional characteristics of these cells, with particular reference to the properties of antigen presenting cells and lymphocytes. ICs were isolated from sponge exudates at days 1-21 after implantation. Dendritic cells (DCs) and lymphocytes were purified using a combination of separation gradients, adherence to plastics, and immunomagnetic sorting. We showed that dominant population of ICs at day 1 and 3 were granulocytes, followed by their decrease thereafter. The number of mononuclear cells (MNCs), macrophages and DCs, progressively increased after day 3, reaching maximal values at day 7. Maximal number of lymphocytes was detected at day 10. In addition, total ICs isolated from day 7 till day 10 exerted significant suppressive activity in co-culture with autologous ConA-stimulated thymocytes and alogeneic lymph node T cells. This finding correlated with the increased level of IL-10 in culture supernatants and the increased proportion MHC class II+ IDO+ DCs and CD3+ CD25+ Foxp3+ lymphocytes at day 7 and day 10, respectively. We did not find any significant difference in the number of these cell populations between regional lymph nodes MNCs and peripheral blood MNCs of experimental animals compared to controls. In conclusion, accumulation of cells with tolerogenic characteristics in GFBR might be relevant for the suppression of inflammation and the prevention of unwanted immune response.
Front. Immunol. Conference Abstract:15th International Congress of Immunology (ICI), Milan, Italy; 08/2013
[Show abstract][Hide abstract] ABSTRACT: Introduction: Granulocytes are essential mediators of the acute inflammatory response and resolution of infection. Numerous studies showed that the function of these cells in children with cystic fibrosis (CF) is affected severely, although the mechanisms are not elucidated fully. Since Toll-like receptors (TLRs) mediate the microbicidal activity and inflammatory pathology of granulocytes, we wondered whether the TLR signaling in granulocytes of CF children is different from that of healthy donors.
Materials and Methods: Granulocytes isolated from peripheral blood by density gradient, were cultivated either with inactivated Aspergillus conidia (a common infectious agent in CF), TLR agonists (TLR2-zymosan, TLR4-LPS, TLR9-CpG) or N-acetyl-cystein (used in CF therapy) for 2h, following the evaluation of myeloperoxidase activity by modified Graham-Knoll procedure. The apoptosis of granulocytes was evaluated after 24h-cultures by analyzing the chromatin condensation, whereas their cytokine production (IL-8 and IL-6) was assessed by ELISA.
Results: Granulocytes of CF patients stimulated with Aspergillus, LPS, or zymosan had significantly lower myeloperoxidase activity compared to controls, and N-acetyl-cystein additionally diminished their myeloperoxidase activity. The apoptosis of granulocytes of CF patients was more inhibited by TLR stimuli and additionally potentiated by Aspergillus, compared to controls. IL-6 and IL-8 levels produced by granulocytes of CF patients was higher than in control cultures, especially in children with exacerbations or advanced stage of the disease (FEV1 and FVC<30%).
Conclusion: By using different stimuli and inhibitors it might be possible to modify functional activity of granulocytes, and thus provide a potential target for modulating the immune response to Aspergillus fumigates and other pathogens in CF.
[Show abstract][Hide abstract] ABSTRACT: In the present study we investigated the cytochrome C oxidase (CO) activity and nitric oxide synthase (NOS) isoenzyme expression after intrahippocampal AlCl3 application in selective vulnerable brain structures. A single dose of AlCl3 was applied in the CA1 sector of rats hippocampus. For biochemical analysis, the animals were killed 10 min and three days after the treatment and forebrain cortex, basal forebrain and hippocampus were removed. Activity of CO was decreased bilaterally in the AlCl3-treated groups in all examined brain structures. We also applied immunohistochemical techniques to identify changes induced by AlCl3 injection after survival periods of 10 min and three days. Both the nNOS and eNOS stains were detected in the hippocampus of controls and AlCl3-treated animals, but iNOS labelling was present in the hippocampus only three days after AlCl3 application. An increased iNOS expression three days post AlCl3 administration could be involved in the mechanism of CO activity depletion.
[Show abstract][Hide abstract] ABSTRACT: Immunoregulatory mechanisms within periapical lesions (PLs) are as yet unexplored. Considering the crucial role of DCs in controlling the immune response within PLs, the immunomodulatory properties of mesenchymal stem cells (MSCs), and the co-localisation of MSCs and DCs in situ, we wondered whether MSCs from PLs modulate the development and functions of DCs. Using a model of monocyte-derived DCs, we showed that PL-MSCs inhibited differentiation of DCs via soluble factors, of which IL-6 had a minor effect, but did not impair their subsequent maturation induced by pro-inflammatory cytokines. However, upon maturation such DCs favoured the production of Th2/Th17 cytokines by allogenic CD4(+) lymphocytes in co-culture, compared with mature DCs differentiated without PL-MSCs. PL-MSC-differentiated DCs, cultivated with pro-inflammatory cytokines and PL-MSCs, although phenotypically mature, exhibited poor allostimulatory activity, induced anergy, Th2 polarisation, differentiation of suppressive CD4(+) CD25(high) CD39(+) Treg-cell subsets via IDO-1-, ILT-3-, and ILT-4-depended mechanisms, and increased production of TGF-β in the co-culture. In contrast, DCs cultivated with PL-MSCs only during maturation stimulated proliferation and Th1 polarisation of CD4(+) T cells in an IL-12-independent manner. In conclusion, PL-MSCs significantly modulate the development and functions of DCs, depending on the phase of DCs development during which the interaction occurs. This article is protected by copyright. All rights reserved.
European Journal of Immunology 07/2013; 43(7). DOI:10.1002/eji.201243010 · 4.03 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The effect of 10-hydroxy-2-decenoic acid (10-HDA), the unique component of royal jelly, on maturation and functions of human monocyte-derived dendritic cells (MoDCs) in culture was investigated. It has been shown that 10-HDA, at concentrations higher than 500 MM, induces apoptosis of MoDCs. A lower dose (50 AM) stimulated T helper (Th)1 and down-regulated Th2 immune responses, as judged by the levels of interferon-gamma (IFN-gamma) and interleukin (IL)-4, respectively, in supernatants of 10-HDA-treated MoDCs cultivated with allogeneic CD4(+)T cells. In contrast, a higher dose of 10-HDA (500 mu M), although non-cytotoxic, inhibited maturation of lipopolysaccharide (LPS)-stimulated MoDCs. Such treated MoDCs produced lower levels of IL-12, IL-18 and tumour necrosis factor-alpha (TNF-alpha) and down-regulated both Th1 and Th2 immune responses. In conclusion, our results suggest that 10-HDA exerts different activity on human MoDCs, depending on applied concentrations, which is important when considering its therapeutic immunomodulatory property.
[Show abstract][Hide abstract] ABSTRACT: Morbidity and mortality of continous ambulatory peritoneal dialysis (CAPD) patients is still very high. The aim of the study was to evaluate the effects of peritoneal dialysis (PD) solutions (standard vs biocompatible) on long-term patients' and the techique survival.
A total of 42 stable patients on CAPD participated in this cross-sectional study. They were prospectively followed-up during the twelve years. Patients with severe anemia (Hb < 10 g/L) and malignant disease ware excluded. Twenty one (50%/0) patients were treated with the standard PD solutions (CAPDP-1) while the other 21 (500/0) were treated with biocompatible PD solutions [(lower level of glucose degradation products, lower concentration of Ca(2+) and neutral pH (CAPDP-2)]. All patients were analyzed for a presence of vascular calcification, nutrition status, and parameters of inflammation after 2.5 +/- 0.6 years of starting CAPD, and these variables considered in the analysis as risk factors.
The patients from the group CAPDP-2 compared to those from the group CAPDP-1 had lower level of high-sensitivity C-reactive protein (hs-CRP) (p = 0.003), and better nutritional status as confirmed by the mid-arm circumference (p = 0.015), and mid-arm muscle circumference (p = 0.002) and subjective global assessment (p = 0.000). Also, they had lower vascular calcifications as confirmed by intima media thickness (IMT) (p = 0.003), degree of carotid narrowing (p = 0.001) and calcified plaques of common carotid arteries (CCA) (p = 0.008). Kaplan-Meier analysis confirmed better survival of patients from the group CAPDP-2 than those from the group CAPDP-1 (1-, 5-, and 10-year patients survival rate was: 100%, 61.9% and 14.3% for the group CAPDP-1, and 100%, 85.7%, and 52.4% for the group CAPDP-2, respectively; p = 0.0345). The 1-, 5-, and 10-year technique survival rate was: 100%, 71.4%, and 38.1% for the group CAPDP-1, and 100%, 85.7%, and 76.2% for the group CAPDP-2, respectively; (p = 0.0719). Duration of dialysis, serum triglyceride and cardiovascular score (quantitative scoring system consisting of: ejection fraction (EF) of left ventricle < 50%; IMT > 1 mm; carotid narrowing degree > 50%, presence of carotid plaques in both common carotide, ischaemic heart disease, cerebrovascular event and peripheral vascular disease with or without amputation) were independent predictors of overall patient survival. Duration of dialysis was only independent predictor of overall technique survival.
Although patients treated with biocompatible solutions showed significantly better survival, the role of biocompatibility of CAPD solutions in patients and technique survival have to be confirmed. Namely, multivariate analysis confirmed that duration of dialysis, serum triglyceride and cardiovascular score significantly predicted overall CAPD patients survival, while only duration of dialysis was found to be independent predictor of overall techique survival.
[Show abstract][Hide abstract] ABSTRACT: Dendritic cells (DCs) are the most important antigen-presenting cells able to sense any change in the environment and to respond by stimulating or suppressing the whole immune system. Such cells are potentially powerful tool for the cell-based anti-tumour therapy. A potential anti-tumour strategy includes the co-delivery of tumour antigens and specific activators into DCs in vivo via gold nanoparticles (GNPs) as carriers. However, many details related to the GNPs’ cytotoxicity, immunomodulatory properties and mechanisms of internalization by DCs remains to be investigated. Using the light microscopy and flow cytometry, we showed that GNPs of different sizes (10nm, 50nm and 90nm) are internalized easily by immature (i)DC, which represent a good target for DC-based vaccines, in contrast to mature (m)DC. State-of-the-art confocal microscopy, focused ion beam/scanning electron microscopy (FIB/SEM) and TEM revealed that GNPs within iDC are distributed predominantly in the endosomal vesicles, but also in the cytoplasm, pointing to the endosomal escape. Additionally, these techniques, and the use of specific inhibitor of dynamin, showed that 10nm GNPs are more prominent to the endosomal escape. Cutting-edge Proton-Induced X-ray Emission Spectroscopy (PIXE)-based quantification by the thin sample approximation (Ogrnic, N et al., ICNMTA 2013, doi: 10.1016/j.nimb.2012.12.060), showed that the mass of GNPs internalized by DC increase with GNPs diameter, but it is inversely proportional to their number within the cells. In conclusion, the choice of GNPs for DCs targeting should be based on both size-dependent internalization, as well as their intracellular behaviour, both of which should maximize the delivery and avoid potentially adverse reactions.