The present study was designed to evaluate the possible effects of the paediatric vaccination schedule in the United States on the central nervous system in a murine model. We compared the impact of treatment with the whole vaccines versus true placebo control. Seventy-six pups were divided into three groups: two vaccinated groups and unvaccinated control. The two vaccinated groups were treated between 7 and 21 post-natal days either with one or three times of the vaccine doses per body weight as used in children between newborn and eighteen months of age. The post-vaccination development, neuromotor behaviours and neurobehavioural abnormalities (NBAs) were evaluated in all mouse groups during the 67 post-natal weeks of mouse age. Mouse body weight was affected only in the vaccinated females compared to males and control. Some NBAs such as decreased sociability, increased anxiety-like behaviours, and alteration of visual-spatial learning and memory were observed in vaccinated male and female mice compared to controls. The present study also shows a slower acquisition of some neonatal reflexes in vaccinated female mice compared to vaccinated males and controls. The observed neurodevelopmental alterations did not show a linear relationship with vaccine dose, suggesting that the single dose gave a saturated response. The outcomes seemed to be sex-dependent and transient with age.

McIntyre Powder (MP) is a finely ground aluminum powder that was used between 1943 and 1979 as a prophylaxis for silicosis. Silicosis is a chronic lung disease caused by the inhalation of crystalline silica dust and was prevalent in the Canadian mining industry during this time period. The McIntyre Research Foundation developed, patented, and produced the MP and distributed it to licensees in Canada, the United States, Mexico, Chile, Belgian Congo, and Western Australia. In the province of Ontario, Canada it is estimated that at least 27,500 miners between 1943 and 1979 were exposed to MP. The present study was undertaken to examine the chemical and physical characteristics of two variations of MP (light grey and black). Chemical analyses (using X-ray Fluorescence and Inductively Coupled Plasma approaches) indicate that the black MP contains significantly higher concentrations of aluminum and metal impurities than the light grey MP (p < 0.001). X-ray diffractometry shows that while aluminum hydroxide dominates the aluminum speciation in both variations, the higher total aluminum content in the black MP is attributable to a greater proportion of elemental aluminum. Physical characterization (using electron microscopy, light microscopy, and dynamic light scattering) indicates that the light grey MP consists of particles ranging from 5 nm to 5 µm in diameter. Atomic Force Microscopy shows that the light grey MP particles in the nanoparticle range (<100 nm) have a mode between 5 and 10 nm. Consequently, it is possible that inhaled smaller MP nanoparticles may be transported via blood and lymph fluid circulation to many different organs including the brain. It is also possible for inhaled larger MP particles to deposit onto lung tissue and for potential health effects to arise from inflammatory responses through immune activation. This MP characterization will provide crucial data to help inform future toxicological, epidemiological, and biological studies of any long-term effects related to the inhalation of aluminum dust and nanomaterials.

Shawqi H Alawdi 1,2 Housam Eidi Marwa M Safar Mosaad A Abdel-Wahhab 6 1Department of Pharmacology, Faculty of Medicine and Health Sciences, Thamar University, Dhamar, Yemen; 2Department of Pharmacy Practice, Faculty of Pharmacy, University of Science and Technology, Sana’a, Yemen; 3Department of Ophthalmology and Visual Sciences, University of British Columbia, Vancouver, Canada; 4Pharmacology and Toxicology Department, Faculty of Pharmacy, Cairo University, Cairo, Egypt; 5Pharmacology and Biochemistry Department, Faculty of Pharmacy, The British University in Egypt, El Shorouk, Egypt; 6Department of Food Toxicology and Contaminants, National Research Center, Cairo, Egypt Background: Diamond nanoparticles (Nanodiamond) are biocompatible drug delivery platforms with outstanding surface properties. Their passage into the brain has been confirmed previously. Thus, nanodiamond could provide a drug delivery system to shuttle several drugs through the blood-brain barrier (BBB) which represents a real challenge for the effective delivery of several drugs into the brain. Amlodipine is a calcium channel blocker that cannot pass through BBB and may elicit neuroprotective effects to reverse calcium-induced excitotoxicity and mitochondrial dysfunction that underlie several neurologic disorders including Alzheimer’s disease and stroke. Aim: The study aimed to investigate the loading of amlodipine on nanodiamond particles. Methods: Nanodiamond particles were oxidized in a strong oxidizing acidic mixture of sulfuric and nitric acids. Adsorption of amlodipine on nanodiamond particles was achieved in alkaline pH using various concentrations of sodium hydroxide. The loaded amlodipine was determined by high-performance liquid chromatography and confirmed by Fourier transform infrared (FTIR) spectroscopy and transmission electron microscopy. Results: The highest percentage (41%) of loaded amlodipine onto nanodiamond particles was achieved in alkaline medium using 2 mM NaOH at a corresponding pH of 8.5. Also, characteristic FTIR bands of amlodipine and nanodiamond were shown obviously in the nanodiamond–amlodipine conjugates. Moreover, the successful loading of amlodipine on diamond nanoparticles was confirmed by transmission electron microscopy. Conclusion: The present study demonstrates the successful loading of amlodipine onto nanodiamond particles. These findings offer a potential for applying diamond nanoparticles as a drug delivery system to shuttle amlodipine into the brain and open the door to deliver other similar drugs into the brain.

Influence of heat or cold stress in sedated animals is unclear and requires further investigations. The present study aimed to evaluate the season's effects on some clinical, hematological parameters and blood cortisol level in sedated Arabian horses with xylazine. Therefore, seven Arabian horses were used to investigate heart and respiratory rates, and capillary refill time and serum cortisol level were recorded before (0) and at 5, 15, 60, and 180 minutes postsedation. Heparinized venous samples were collected before (0) and 3 hours postsedation for analysis of hematological analysis. Arterial blood samples were collected before and 1 hour postsedation for arterial blood gases and electrolytes analysis. Repeated analysis of variance was performed (P < .05). Significant decreases have been observed in heart and respiratory rates at 5, 15, and 60 minutes postsedation in summer and only at 5 minutes postsedation in winter. Arterial oxygen pressure and arterial carbon dioxide pressure showed a significant decrease and increase, respectively at 1 hour postsedation in summer and winter. The serum cortisol levels were significantly higher in summer than in winter at 5, 15, and 60 minutes postsedation. In summer, the postsedation concentrations of cortisol did not change significantly than its values before sedation. However, in winter, the cortisol concentration decreased significantly at 5, 15, and 60 minutes postsedation compared with their value before sedation. The present study suggests that these season's effects on the sedated Arabian horses could take into consideration in xylazine-sedated Arabian horses.

BACKGROUND:Diamond nanoparticles (Nanodiamond) are biocompatible drug delivery platforms with outstanding surface properties. Their passage into the brain has been confirmed previously. Thus, nanodiamond could provide a drug delivery system to shuttle several drugs through the blood-brain barrier (BBB) which represents a real challenge for the effective delivery of several drugs into the brain. Amlodipine is a calcium channel blocker that cannot pass through BBB and may elicit neuroprotective effects to reverse calcium-induced excitotoxicity and mitochondrial dysfunction that underlie several neurologic disorders including Alzheimer's disease and stroke.AIM:The study aimed to investigate the loading of amlodipine on nanodiamond particles.METHODS:Nanodiamond particles were oxidized in a strong oxidizing acidic mixture of sulfuric and nitric acids. Adsorption of amlodipine on nanodiamond particles was achieved in alkaline pH using various concentrations of sodium hydroxide. The loaded amlodipine was determined by high-performance liquid chromatography and confirmed by Fourier transform infrared (FTIR) spectroscopy and transmission electron microscopy.RESULTS:The highest percentage (41%) of loaded amlodipine onto nanodiamond particles was achieved in alkaline medium using 2 mM NaOH at a corresponding pH of 8.5. Also, characteristic FTIR bands of amlodipine and nanodiamond were shown obviously in the nanodiamond-amlodipine conjugates. Moreover, the successful loading of amlodipine on diamond nanoparticles was confirmed by transmission electron microscopy.CONCLUSION:The present study demonstrates the successful loading of amlodipine onto nanodiamond particles. These findings offer a potential for applying diamond nanoparticles as a drug delivery system to shuttle amlodipine into the brain and open the door to deliver other similar drugs into the brain.

Abstract Introduction: Vitamin D (VD) deficiency is a major public health problem worldwide. In spite of its high prevalence, particularly among elderly people, VD deficiency is still underestimated by many physicians. Increasingly, VD deficiency is associated with several known geriatric syndromes. Methods: The study sample consisted of 125 patients, aged 75 years and older, admitted to the acute geriatric unit. The plausible association between the serum 25-hydroxyvitamin D [25(OH)D] level and patient age, sex, body mass index, renal function, cholecystectomy history, and the prescribed drugs had been investigated. The Fisher's exact test was used to conduct the statistical analysis of data. Results: Surprisingly, furosemide treatment was correlated with normal 25(OH)D levels and an increased incidence of secondary hyperparathyroidism. Unlike the other four parameters mentioned above, our data showed that only the patient sex exhibited a significant association with 25(OH)D level as elderly males suffered from a serious VD deficiency as compared to elderly females. Conclusion: Old age is an independent risk factor for VD deficiency. The supplementary dose of VD should be precisely defined to achieve the optimal serum 25(OH)D level in elderly people. The definition of the normal serum 25(OH)D threshold in elderly furosemide-treated patients is worth of further studies.

The use of nanoparticles (NPs) seems to make life easier. In medicine, for instance, nanoparticles are used for targeted treatments, either by surface targeting or by magnetic guidance. This is done by manipulating the size and characteristics of the particles while choosing the matrix constituents. Mitochondrial depolarization can induce lysosomal mitochondria alteration using a process called mitophagy. Mitophagy plays an essential role in mitochondrial homeostasis, regulating their size and quality. Mitophagy eliminates damaged mitochondria, which could be induced under diverse stress conditions including pathogens and biopersistent NPs. NPs can affect autophagy by signaling pathways or by gene/protein expression. Autophagy induction by NPs could be considered a degradation process of foreign or aberrant agents for cells, such as bacteria and virus. A better insight is also needed on how a cellular response could affect the final fate of NPs by providing a different route to transport NPs between distinct types of cell organelles.

Current therapeutic approaches of Alzheimer’s disease (AD) are symptomatic and of modest efficacy, and there is no available effective cure or prevention of AD; hence, the need arise to search for neuroprotective agents to combat AD. The current study aimed at investigating the neuroprotective effect of nanodiamond (ND), adamantine-based nanoparticles, in aluminum-induced cognitive impairment in rats, an experimental model of AD. AD was induced by aluminum chloride (17 mg/kg, p.o. for 6 weeks) and confirmed by Morris water maze and Y-maze behavioral tests. Biochemical and histological analyses of the hippocampus were also performed. Aluminum-treated rats showed behavioral, biochemical, and histological changes similar to those associated with AD. ND improved learning and memory and reversed histological alterations. At the molecular levels, ND mitigated the increase of hippocampal beta-amyloid (Aβ42) and beta-site amyloid precursor protein cleaving enzyme-1 (BACE1) together with down-regulation of phosphorylated tau protein. It also modulated the excitatory glutamate neurotransmitter level. Furthermore, ND boosted the brain-derived neurotrophic factor (BDNF) and mitochondrial transcription factor-A (TFAM), suppressed the proinflammatory cytokine tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), and curbed oxidative stress by hampering of inducible nitric oxide synthase (iNOS). Moreover, ND augmented the hippocampal levels of phosphorylated signal transducer and activator of transcription-3 (p-STAT3) and B cell leukemia/lymphoma-2 (Bcl-2) anti-apoptotic protein while diminished nuclear factor-kappaB (NF-κB) and caspase-3 (casp-3) expression. These findings indicate the protective effect of ND against memory deficits and AD-like pathological aberrations probably via modulating NF-kB and STAT3 signaling, effects mediated likely by modulating N-methyl-D-aspartate (NMDA) receptors.

Aluminium hydroxide (alum) has long been added as an adjuvant of vaccines. It consists of nanoparticles forming aggregates. Unexpectedly long-lasting biopersistence of alum aggregates were found within immune cells of patients with chronic fatigue, cognitive dysfunction, myalgias and dysimmunity [1], [2]. We documented in mice slow translocation of alum aggregates captured by monocyte-lineage cells from the injected muscle to brain [3], [4], [5]. Herein, brain function and aluminium (Al) concentration were examined long after injections. Alhydrogel® was injected in TA muscle in adult female CD1 mice at 3 doses ranging from 133 to 800 μg Al/kg. Eight validated tests were used to evaluate cognitive and motor performances 180 days after injection. Brains were collected for Al level determination and Iba-1 immunohistochemistry. A most unusual neuro-toxicological pattern limited to lower doses of alum was observed. Neurobehavioral changes, including decreased activity levels and altered anxiety-like behaviour, were documented in animals exposed to the two lowest doses (133 and 200 μg Al/kg) but not at the highest dose (800 μg Al/kg), compared to controls. Consistently, cerebral Al levels were increased in animals exposed to the lowest doses. Microglial cell increase was found in amygdala of the 200 μg Al/kg group. Interestingly, the injected suspensions corresponding to the two lowest doses contained much smaller aggregates (1.50–1.75 μm) compared to the highest dose (4.70 μm). Alum particles injected in muscle may induce neurotoxic effects and Al cerebral accumulation six months after injection in mice. Neurotoxic effects are restricted to low concentration suspensions forming small particle aggregates. Such bacteria-sized aggregates are known to be selectively captured by monocyte-lineage cells. This study strongly suggests that, in contrast to “the dose makes the poison” paradigm of classical toxicology, alum toxicology obeys the specific rules of small particle toxicology, thus deserving in depth revaluation. (This study was supported by ANSM).

Aluminium (Al) oxyhydroxide (Alhydrogel®), the main adjuvant licensed for human and animal vaccines, consists of primary nanoparticles that spontaneously agglomerate. Concerns about its safety emerged following recognition of its unexpectedly long-lasting biopersistence within immune cells in some individuals, and reports of chronic fatigue syndrome, cognitive dysfunction, myalgia, dysautonomia and autoimmune/inflammatory features temporally linked to multiple Al-containing vaccine administrations. Mouse experiments have documented its capture and slow transportation by monocyte-lineage cells from the injected muscle to lymphoid organs and eventually the brain. The present study aimed at evaluating mouse brain function and Al concentration 180days after injection of various doses of Alhydrogel® (200, 400 and 800μg Al/kg of body weight) in the tibialis anterior muscle in adult female CD1 mice. Cognitive and motor performances were assessed by 8 validated tests, microglial activation by Iba-1 immunohistochemistry, and Al level by graphite furnace atomic absorption spectroscopy. An unusual neuro-toxicological pattern limited to a low dose of Alhydrogel® was observed. Neurobehavioural changes, including decreased activity levels and altered anxiety-like behaviour, were observed compared to controls in animals exposed to 200μg Al/kg but not at 400 and 800μg Al/kg. Consistently, microglial number appeared increased in the ventral forebrain of the 200μg Al/kg group. Cerebral Al levels were selectively increased in animals exposed to the lowest dose, while muscle granulomas had almost completely disappeared at 6 months in these animals. We conclude that Alhydrogel® injected at low dose in mouse muscle may selectively induce long-term Al cerebral accumulation and neurotoxic effects. To explain this unexpected result, an avenue that could be explored in the future relates to the adjuvant size since the injected suspensions corresponding to the lowest dose, but not to the highest doses, exclusively contained small agglomerates in the bacteria-size range known to favour capture and, presumably, transportation by monocyte-lineage cells. In any event, the view that Alhydrogel® neurotoxicity obeys "the dose makes the poison" rule of classical chemical toxicity appears overly simplistic.

Concerns regarding vaccine safety have emerged following reports of potential adverse events in both humans and animals. In the present study, alum, alum-containing vaccine and alum adjuvant tagged with fluorescent nanodiamonds were used to evaluate i) the persistence time at the injection site, ii) the translocation of alum from the injection site to lymphoid organs, and iii) the behavior of adult CD1 mice following intramuscular injection of alum (400 μg Al/kg). Results showed for the first time a strikingly delayed systemic translocation of adjuvant particles. Alum-induced granuloma remained for a very long time in the injected muscle despite progressive shrinkage from day 45 to day 270. Concomitantly, a markedly delayed translocation of alum to the draining lymph nodes, major at day 270 endpoint, was observed. Translocation to the spleen was similarly delayed (highest number of particles at day 270). In contrast to C57BL/6J mice, no brain translocation of alum was observed by day 270 in CD1 mice. Consistently neither increase of Al cerebral content, nor behavioral changes were observed. On the basis of previous reports showing alum neurotoxic effects in CD1 mice, an additional experiment was done, and showed early brain translocation at day 45 of alum injected subcutaneously at 200 μg Al/kg. This study confirms the striking biopersistence of alum. It points out an unexpectedly delayed diffusion of the adjuvant in lymph nodes and spleen of CD1 mice, and suggests the importance of mouse strain, route of administration, and doses, for future studies focusing on the potential toxic effects of aluminum-based adjuvants.

Aluminum oxyhydroxide (alum) is a crystalline compound widely used as an immunological adjuvant of vaccines. Concerns linked to the use of alum particles emerged following recognition of their causative role in the so-called macrophagic myofasciitis (MMF) lesion detected in patients with myalgic encephalomyelitis/chronic fatigue/syndrome. MMF revealed an unexpectedly long-lasting biopersistence of alum within immune cells in presumably susceptible individuals, stressing the previous fundamental misconception of its biodisposition. We previously showed that poorly biodegradable aluminum-coated particles injected into muscle are promptly phagocytosed in muscle and the draining lymph nodes, and can disseminate within phagocytic cells throughout the body and slowly accumulate in brain. This strongly suggests that long-term adjuvant biopersistence within phagocytic cells is a prerequisite for slow brain translocation and delayed neurotoxicity. The understanding of basic mechanisms of particle biopersistence and brain translocation represents a major health challenge, since it could help to define susceptibility factors to develop chronic neurotoxic damage. Biopersistence of alum may be linked to its lysosome-destabilizing effect, which is likely due to direct crystal-induced rupture of phagolysosomal membranes. Macrophages that continuously perceive foreign particles in their cytosol will likely reiterate, with variable interindividual efficiency, a dedicated form of autophagy (xenophagy) until they dispose of alien materials. Successful compartmentalization of particles within double membrane autophagosomes and subsequent fusion with repaired and re-acidified lysosomes will expose alum to lysosomal acidic pH, the sole factor that can solubilize alum particles. Brain translocation of alum particles is linked to a Trojan horse mechanism previously described for infectious particles (HIV, HCV), that obeys to CCL2, signaling the major inflammatory monocyte chemoattractant.

BACKGROUND:Aluminum oxyhydroxide (alum) is a crystalline compound widely used as an immunologic adjuvant of vaccines. Concerns linked to alum particles have emerged following recognition of their causative role in the so-called macrophagic myofasciitis (MMF) lesion in patients with myalgic encephalomyelitis, revealing an unexpectedly long-lasting biopersistence of alum within immune cells and a fundamental misconception of its biodisposition. Evidence that aluminum-coated particles phagocytozed in the injected muscle and its draining lymph nodes can disseminate within phagocytes throughout the body and slowly accumulate in the brain further suggested that alum safety should be evaluated in the long term. However, lack of specific staining makes difficult the assessment of low quantities of bona fide alum adjuvant particles in tissues.METHODS:We explored the feasibility of using fluorescent functionalized nanodiamonds (mfNDs) as a permanent label of alum (Alhydrogel(®)). mfNDs have a specific and perfectly photostable fluorescence based on the presence within the diamond lattice of nitrogen-vacancy centers (NV centers). As the NV center does not bleach, it allows the microspectrometric detection of mfNDs at very low levels and in the long-term. We thus developed fluorescent nanodiamonds functionalized by hyperbranched polyglycerol (mfNDs) allowing good coupling and stability of alum:mfNDs (AluDia) complexes. Specificities of AluDia complexes were comparable to the whole reference vaccine (anti-hepatitis B vaccine) in terms of particle size and zeta potential.RESULTS:In vivo, AluDia injection was followed by prompt phagocytosis and AluDia particles remained easily detectable by the specific signal of the fND particles in the injected muscle, draining lymph nodes, spleen, liver and brain. In vitro, mfNDs had low toxicity on THP-1 cells and AluDia showed cell toxicity similar to alum alone. Expectedly, AluDia elicited autophagy, and allowed highly specific detection of small amounts of alum in autophagosomes.CONCLUSIONS:The fluorescent nanodiamond technology is able to overcome the limitations of previously used organic fluorophores, thus appearing as a choice methodology for studying distribution, persistence and long-term neurotoxicity of alum adjuvants and beyond of other types of nanoparticles.

The aim of the present work was to functionalize fluorescent nanodiamond by covalent grafting with hyperbranched polyglycerol. Fluorescent nanodiamond, derived from high pressure high temperature micron-sized diamond, was oxidized and then thermally reacted with pure glycidol in the absence of catalyst. The results revealed that thermal polymerization of glycidol was notably faster on the nanodiamond surface as a result of a surface initiation of the isothermal ring opening polymerization. Interestingly, the aqueous dispersion of the resulting nanoparticles appeared stable at high ionic strength. Furthermore, the fluorescent nanodiamond grafted with hyperbranched polyglycerol displayed several hydroxyl end-groups which could be further derivatized by carboxylation or carbamatization and subsequently conjugated with protein linked via an amide bound. Notably, nanodiamonds retain their unique fluorescent characteristics. This work suggests that fluorescent nanodiamond coated with hyperbranched glycidol could be promising in biomedical research where aqueous dispersion of fluorescent nanoparticles stable in physiological medium is in high demand to label, track and quantify biomolecules.

Due to their unique properties, engineered nanoparticles (NPs) have found broad use in industry, technology, and medicine, including as a vehicle for drug delivery. However, the understanding of NPs' interaction with different types of mammalian cells lags significantly behind their increasing adoption in drug delivery. In this study, we show unique responses of human epithelial breast cells when exposed to polymeric Eudragit((R)) RS NPs (ENPs) for 1-3 days. Cells displayed dose-dependent increases in metabolic activity and growth, but lower proliferation rates, than control cells, as evidenced in tetrazolium salt (WST-1) and 5-bromo-2'-deoxyuridine (BrdU) assays, respectively. Those effects did not affect cell death or mitochondrial fragmentation. We attribute the increase in metabolic activity and growth of cells culture with ENPs to three factors: (1) high affinity of proteins present in the serum for ENPs, (2) adhesion of ENPs to cells, and (3) activation of proliferation and growth pathways. The proteins and genes responsible for stimulating cell adhesion and growth were identified by mass spectrometry and Microarray analyses. We demonstrate a novel property of ENPs, which act to increase cell metabolic activity and growth and organize epithelial cells in the epithelium as determined by Microarray analysis.

Gold nanoparticles (AuNP) hold great potential for biomedical applications. This study was aimed at examination of the effect of AuNP coating on the redox status of their environment. Two kinds of AuNP were tested, similar by shape and size, but with different surface coatings: either stabilized with citrate or functionalized with dihydrolipoic acid (Au@DHLA NP). Interestingly, whereas citrate-stabilized AuNP interact in vitro with reduced glutathione (GSH) and S-nitrosoglutathione, Au@DHLA NP do not interfere with both biomolecules. Albumin exhibits higher affinity toward citrate-stabilized AuNP than Au@DHLA NP, increasing their hydrodynamic diameter (8.0- and 1.3-fold, respectively). Furthermore, the AuNP coating affects also their internalization by macrophages (which was two fold higher for citrate-stabilized AuNP), following an exposure to a subtoxic NP concentration (10 nM, 80% viability). Citrate-stabilized AuNP were found to decrease the intracellular GSH level (ca. 20%), with no increase in reactive oxygen species production. Furthermore, these AuNP did not induce apoptosis (as shown by caspase-3 activity and nfkb2 transcription factor), and also did not activate gene expression related to oxidative stress (ncf1) and inflammatory response (tnfα). The present data highlight that the functionalization of AuNP with DHLA decreases their reactivity with biomolecules and cells, resulting in a promising medical platform.

Taxanes are potent antimitotic agents that have demonstrated efficacy in a wide range of malignancies. Due to their poor water-solubility, these cytostatic drugs were first formulated with low molecular weight surfactants, e.g. Cremophor® EL (CrEL) and Tween 80® (polysorbate 80), which are known to exhibit serious adverse effects in humans. In recent years, there has been growing interest in the design of more biocompatible formulations for both paclitaxel and docetaxel. Polymer-based drug carriers represent an attractive venue given the diversity in the array of existing polymers. Most notably, biopolyesters are vastly employed in the field of biomedical research given their biocompatibility and biodegradability. Polyester-based micelles and nanoparticles have been applied to the parenteral delivery of taxanes with varying degrees of success. Block copolymer micelles possess a unique core-shell structure generated through the self-assembly of amphiphilic copolymers in aqueous media. Although these systems have shown greatly enhanced tolerability compared to formulations based on low molecular weight surfactants, in some cases their failure to retain their cargo following parenteral administration has hindered their capacity to target taxanes to solid tumours. While polyester-based nanoparticles possess comparatively greater stability and drug targeting capacity, they frequently display a significant burst effect whereby a major portion of the cargo is immediately discarded from the carrier upon injection. This review focuses on the current application of polyester-based micelles and nanoparticles to the tumour targeting of taxanes. The preparation, loading efficiencies, release kinetics, cytotoxicity and in vivo behaviour of these systems is discussed in detail.

L'utilisation de nanoparticules (NPs) comme vecteur de médicaments est actuellement en plein développement de vectoriser des principes actifs dans l'organisme pour en diminuer les effets secondaires. Cependant peu d'études portent sur l'activité biologique de ces NPs et plus particulièrement sur leur toxicité intrinsèque. Récemment, une formulation orale d'Héparine de Bas Poids Moléculaire à base de NPs composées d'un mélange de deux polymères, polycaprolactone et Eudragit® RS, a été développée au sein de l'EA3452. Nous avons entrepris d'étudier la cytotoxicité de ces NPs vides ainsi que celles chargées avec le médicament par deux méthodes d'encapsulation : la nanopréciptation et la double émulsion. La toxicité observée des NP vides a été étudiée. Elles sont toxiques pour la lignée de macrophages de rat NR8383 d'une manière dose- et temps-dépendante. La microscopie électronique a montré que les NPs pénétraient dans la cellule, de façon unitaire, par endocytose et gagnent les mitochondries déclenchant un phénomène de mitophagie alors qu'aucune image évoquant une apoptose n'a été observée. Ceci est confirmé par l'étude des voies métaboliques par « microchip arrays » et RT-PCR quantitative. Les voies métaboliques conduisant à l'autophagie sont activées (en particulier le gène atg1611) sans que l'apoptose soit mise en jeu. La désorganisation des structures mitochondriales est associée à une répression de l'expression du gène opa1. Ces résultats obtenus doivent être confirmé sur des macrophages humains. Cependant ils tendent à montrer que les NPs seules ont des effets biologiques qui doivent être pris en compte avant toute utilisation chez l'homme

Drug delivery nanosystems are currently used in human therapy. In preliminary studies we have observed that Eudragit® RS nanoparticles, prepared by nanoprecipitation or double emulsion techniques, are cytotoxic for NR8383 rat macrophages. In this study, we expand our previous analysis and suggest that unloaded Eudragit® RS nanoparticles prepared by nanoprecipitation (NP/ERS) may induce important morphological and biochemical cellular modifications leading to cellular death. In NR8383 rat macrophages cell line exposed to doses varying from 15 to 100μg/mL, NP/ERS nanoparticles are internalized inside the cells, reach the mitochondria and alter the structure of these organelles. In addition, the exposure to nanoparticles induces cellular autophagy as demonstrated by electron microscopy analysis, microchip array, qRT-PCR and Western blot assays. Although toxicity of nanoparticles has already been evidenced, it is the first time that results show clearly that the toxicity of polymeric nanovectors may be related to an activation of autophagy.