Co-encapsulation of fisetin and cisplatin into liposomes: Stability considerations and in vivo efficacy on lung cancer animal model.

Lung cancer is a highly vascularized tumor for which a combination between an antitumor agent, cisplatin, and an antiangiogenic molecule, fisetin, appears a promising therapeutic approach. In order to deliver both chemotherapies within the tumor, to enhance fisetin solubility and decrease cisplatin toxicity, an encapsulation of both drugs into liposomes was developed. Purification and freeze-drying protocols were optimized to improve both the encapsulation and liposome storage. The cytotoxicity of the encapsulated chemotherapies was evaluated on Lewis lung carcinoma (3LL) cell lines. The antitumor effect of the combination was evaluated in vivo on an ectopic mouse model of Lewis Lung carcinoma. The results showed that fisetin and cisplatin co-loaded liposomes were successfully prepared. Freeze-drying allowed a 30 days storage limiting the release of both drugs. The combination index between liposomal fisetin and liposomal cisplatin on 3LL cell line after 24 h of exposure showed a clear synergism: CI = 0.7 for the co loaded liposomes and CI = 0.9 for the mixture of cisplatin loaded and fisetin loaded liposomes. The co-encapsulating formulation showed in vivo efficacy against an ectopic murine model of Lewis Lung carcinoma with a probable reduction in the toxicity of cisplatin through co-encapsulation with fisetin.

H2 O2 -Induced Persistent Luminescence Signal Enhancement Applied to Biosensing.

Persistent luminescence nanoparticles (PLNPs) are innovative materials able to emit light for a long time after the end of their excitation. Thanks to this property, their detection can be separated in time from the excitation, making it possible to obtain images with a high signal-to-noise ratio. This optical property can be of particular interest for the development of in vitro biosensors. Here, we report the unexpected effect of hydrogen peroxide (H2 O2 ) on the signal intensity of ZnGa2 O4 :Cr3+ (ZGO) nanoparticles. In the presence of H2 O2 , the signal intensity of ZGO can be amplified. This signal amplification can be used to detect and quantify H2 O2 in various media, using non-functionalized ZGO nanoparticles. This small molecule can be produced by several oxidases when they react with their substrate. Indeed, the quantification of glucose, lactic acid, and uric acid is possible. The limit of detection could be lowered by modifying the nanoparticles synthesis route. These optimized nanoparticles can also be used as new biosensor to detect larger molecules such as antigen, using the appropriate antibody. This unique property, i.e., persistent luminescence signal enhancement induced by H2 O2 , represents a new way to detect biomolecules which could lead to a very large number of bioassay applications.

Effect of the Elaboration Method on Structural and Optical Properties of Zn1.33Ga1.335Sn0.33O4:0.5%Cr3+ Persistent Luminescent Nanomaterials.

Near-infrared (NIR) persistent luminescence (PersL) materials have demonstrated promising developments for applications in many advanced fields due to their unique optical properties. Both high-temperature solid-state (SS) or hydrothermal (HT) methods can successfully be used to prepare PersL materials. In this work, Zn1.33Ga1.34Sn0.33O4:0.5%Cr3+ (ZGSO:0.5%Cr3+), a newly proposed nanomaterial for bioimaging, was prepared using SS and HT methods. The results show the crystal structure, morphology and optical properties of the samples that were prepared using both methods. Briefly, the crystallite size of the ZGSO:0.5%Cr3+ prepared using the SS method is ~3 µm, and as expected, is larger than materials prepared using the HT method. However, the growth process used in the hydrothermal environment promotes the formation of ZGSO:0.5%Cr3+ with more uniform shapes and smaller sizes (less than 500 nm). Different diameter ranges of nanoparticles were obtained using HT and ball milling (BM) methods (ranging from 25-50 nm) and by using SS and BM methods (25-200 nm) as well. In addition, the SS-prepared microstructure material has stronger PersL than HT-prepared particles before they go through ball milling to create nanomaterials. On the contrary, after BM treatment, ZGSO:0.5%Cr3+ HT and BM NPs present higher PersL and photoluminescence (PL) properties than ZGSO:0.5%Cr3+ SS and BM NPs, even though both kinds of NPs present worse PersL and PL compared to the original particles before BM. To summarize: preparation methods, whether by SS or HT, with additional grinding as a second step, can have a significant impact on the morphological and luminescent features of ZGSO:0.5%Cr3+ PersL materials.

New Lidocaine-Based Pharmaceutical Cocrystals: Preparation, Characterization, and Influence of the Racemic vs. Enantiopure Coformer on the Physico-Chemical Properties.

This study describes the preparation, characterization, and influence of the enantiopure vs. racemic coformer on the physico-chemical properties of a pharmaceutical cocrystal. For that purpose, two new 1:1 cocrystals, namely lidocaine:dl-menthol and lidocaine:d-menthol, were prepared. The menthol racemate-based cocrystal was evaluated by means of X-ray diffraction, infrared spectroscopy, Raman, thermal analysis, and solubility experiments. The results were exhaustively compared with the first menthol-based pharmaceutical cocrystal, i.e., lidocaine:l-menthol, discovered in our group 12 years ago. Furthermore, the stable lidocaine/dl-menthol phase diagram has been screened, thoroughly evaluated, and compared to the enantiopure phase diagram. Thus, it has been proven that the racemic vs. enantiopure coformer leads to increased solubility and improved dissolution of lidocaine due to the low stable form induced by menthol molecular disorder in the lidocaine:dl-menthol cocrystal. To date, the 1:1 lidocaine:dl-menthol cocrystal is the third menthol-based pharmaceutical cocrystal, after the 1:1 lidocaine:l-menthol and the 1:2 lopinavir:l-menthol cocrystals reported in 2010 and 2022, respectively. Overall, this study shows promising potential for designing new materials with both improved characteristics and functional properties in the fields of pharmaceutical sciences and crystal engineering.

ZGSO Spinel Nanoparticles with Dual Emission of NIR Persistent Luminescence for Anti-Counterfeiting Applications.

The property of persistent luminescence shows great potential for anti-counterfeiting technology and imaging by taking advantage of a background-free signal. Current anti-counterfeiting technologies face the challenge of low security and the inconvenience of being limited to visible light emission, as emitters in the NIR optical windows are required for such applications. Here, we report the preparation of a series of Zn1+xGa2-2xSnxO4 nanoparticles (ZGSO NPs) with persistent luminescence in the first and second near-infrared window to overcome these challenges. ZGSO NPs, doped with transition-metal (Cr3+ and/or Ni2+) and in some cases co-doped with rare-earth (Er3+) ions, were successfully prepared using an improved solid-state method with a subsequent milling process to reach sub-200 nm size particles. X-ray diffraction and absorption spectroscopy were used for the analysis of the structure and local crystal field around the dopant ions at different Sn4+/Ga3+ ratios. The size of the NPs was ~150 nm, measured by DLS. Doped ZGSO NPs exhibited intense photoluminescence in the range from red, NIR-I to NIR-II, and even NIR-III, under UV radiation, and showed persistent luminescence at 700 nm (NIR-I) and 1300 nm (NIR-II) after excitation removal. Hence, these NPs were evaluated for multi-level anti-counterfeiting technology.

Designing fisetin nanocrystals for enhanced in cellulo anti-angiogenic and anticancer efficacy.

We report the formulation, characterization, colloidal stability, and in vitro efficiency of Fisetin nanocrystals stabilized by poloxamer P407. Such nanocrystals present a nanometer scale (148.6 ± 1.1 nm) and a high homogeneity (polydispersity index of 0.17 ± 0.01), with a production yield of 97.0 ± 2.5%. The engineered formulations of nanocrystals suspension (pH of 7.4 ± 0.1), stabilized via steric repulsion, are stable for several days in aqueous environment (Milli Q water, NaCl 10 mM or mannitol 5% w/v), for few days in HEPES buffered saline (HBS) (20 / 150 mM) under sink conditions, and in culture medium. After freeze drying in 5% w/v mannitol, the nanocrystal formulations can be stored at -80 °C for at least 120 days. Drug release experiments displayed a 98.7 ± 5.1% cumulative release over 3 days in HBS. Compared to the free drug, the nanocrystal formulations showed an improved cytotoxicity highlighted by the decrease of the half maximal inhibitory concentration for both murine Lewis lung carcinoma (3LL) and human endothelial (EA.hy926) cell lines. In addition, after incubation with Fisetin nanosuspensions, significant changes in the cell morphology for both cell lines were observed, showing an improved anti-angiogenic effect of nanocrystals formulation compared to the free drug. Overall, Fisetin formulated as nanocrystals showed enhanced biopharmaceutical properties and in vitro activity, offering a wide range of indications for challenging applications in the clinic.

Fate and biological impact of persistent luminescence nanoparticles after injection in mice: a one-year follow-up.

Persistent luminescence nanoparticles (PLNPs) are attracting growing interest for non-invasive optical imaging of tissues with a high signal to noise ratio. PLNPs can emit a persistent luminescence signal through the tissue transparency window for several minutes, after UV light excitation before systemic administration or directly in vivo through visible irradiation, allowing us to get rid of the autofluorescence signal of tissues. PLNPs constitute a promising alternative to the commercially available optical near infrared probes thanks to their versatile functionalization capabilities for improvement of the circulation time in the blood stream. Nevertheless, while biodistribution for a short time is well known, the long-term fate and toxicity of the PLNP's inorganic core after injection have not been dealt with in depth. Here we extend the current knowledge on ZnGa1.995O4Cr0.005 NPs (or ZGO) with a one-year follow-up of their fate after a single systemic administration in mice. We investigated the organ tissue uptake of ZGO with two different coatings and determined their intracellular processing up to one year after injection. The biopersistence of ZGO was assessed, with a long-term retention, quantified by ICP-MS, mostly in the liver and spleen, parallel with a loss of their luminescence properties. The analysis of the toxicity related to combining an animal's weight, key hematological and metabolic markers, histological observations of liver tissues and quantification of the expression of 31 genes linked to different metabolic reactions did not reveal any signs of noxiousness, from the macro scale to the molecular level. Therefore, the ZGO imaging probe has been proven to be a safe and relevant candidate for preclinical studies, allowing its long term use without any in vivo disturbance of the general metabolism.

Persistent X-ray-activated phosphors: mechanisms and applications.

Trivalent lanthanides in wide bandgap fluoride or phosphate hosts can present persistent luminescence between 200 nm and 1.7 µm after charging by X-rays. Mechanisms are reviewed and applications envisioned.

Persistent luminescence nanoparticles functionalized by polymers bearing phosphonic acid anchors: synthesis, characterization, and in vivo behaviour.

Optical in vivo imaging has become a widely used technique and is still under development for clinical diagnostics and treatment applications. For further development of the field, researchers have put much effort into the development of inorganic nanoparticles (NPs) as imaging probes. In this trend, our laboratory developed ZnGa1.995O4Cr0.005 (ZGO) nanoparticles, which can emit a bright persistent luminescence signal through the tissue transparency window for dozens of minutes and can be activated in vivo with visible irradiation. These properties endow them with unique features, allowing us to recover information over a long-time study with in vivo imaging without any background. To target tissues of interest, ZGO must circulate long enough in the blood stream, a phenomenon which is limited by the mononuclear phagocyte system (MPS). Depending on their size, charge and coating, the NPs are sooner or later opsonized and stored into the main organs of the MPS (liver, spleen, and lungs). The NPs therefore have to be coated with a hydrophilic polymer to avoid this limitation. To this end, a new functionalization method using two different polyethylene glycol phosphonic acid polymers (a linear one, later named lpPEG and a branched one, later named pPEG) has been studied in this article. The coating has been optimized and characterized in various aqueous media. The behaviour of the newly functionalized NPs has been investigated in the presence of plasmatic proteins, and an in vivo biodistribution study has been performed. Among them ZGOpPEG exhibits a long circulation time, corresponding to low protein adsorption, while presenting an effective one-step process in aqueous medium with a low hydrodynamic diameter increase. This new method is much more advantageous than another strategy we reported previously that used a two-step PEG silane coating performed in an organic solvent (dimethylformamide) for which the final hydrodynamic diameter was twice the initial diameter.

Electrokinetic elucidation of the interactions between persistent luminescent nanoprobes and the binary apolipoprotein-E/albumin protein system.

The affinity between functional nanoparticles (NPs) and proteins could determine the efficacy of nanoprobes, nanosensors, nanocarriers, and many other devices for biomedical applications. Therefore, it is necessary to develop analytical strategies to accurately evaluate the magnitude of these protein corona interactions in physiological media. In this work, different electrokinetic strategies were implemented to accurately determine the interactions between PEGylated ZnGa1.995Cr0.005O4 persistent luminescent NPs (ZGO-PEG) and two important serum proteins: human serum albumin (HSA), the most abundant serum protein, and apolipoprotein-E (ApoE), associated with the active transport of NPs through the blood-brain barrier. Firstly, the injection of ZGO-PEG in a background electrolyte (BGE) containing individual proteins allowed an affinity study to separately characterize each NP-protein system. Then, the same procedure was applied in a buffer containing a mixture of the two proteins at different molar ratios. Finally, the NPs were pre-incubated with one protein and thereafter electrokinetically separated in a BGE containing the second protein. These analytical strategies revealed the mechanisms (comparative, cooperative or competitive systems) and the magnitude of their interactions, resulting in all cases in notably higher affinity and stability between ZGO-PEG and ApoE (Ka = 1.96 ± 0.25 × 1010 M-M) compared to HSA (Ka = 4.60 ± 0.41 × 106 M-M). For the first time, the inter-protein ApoE/HSA interactions with ZGO-PEG were also demonstrated, highlighting the formation of a ternary ZGO-PEG/ApoE/HSA nanocomplex. These results open the way for a deeper understanding of the protein corona formation, and the development of versatile optical imaging applications for ZGO-PEG and other systemically delivered nanoprobes ideally vectorized to the brain.

Functionalized Multi-Walled Carbon Nanotube-Based Aptasensors for Diclofenac Detection.

Driven by the increasing concern about the risk of diclofenac (DCF) residues as water pollutants in the aqueous environment and the growing need for its trace determination, a simple but sensitive electrochemical aptasensor for the trace detection of DCF was developed. To construct the aptasensor, the amine-terminated DCF aptamer was covalently immobilized on the surface of the carboxylic acid-functionalized multi-walled carbon nanotube (f-MWCNT)-modified glassy carbon electrode (GCE) through EDC/NHS chemistry. The f-MWCNTs provide a reliable matrix for aptamer immobilization with high grafting density, while the aptamer serves as a biorecognition probe for DCF. The obtained aptasensor was incubated with DCF solutions at different concentrations and was then investigated by electrochemical impedance spectroscopy (EIS). It displays two linear ranges of concentration for DCF detection, from 250 fM to 1pM and from 1 pM to 500 nM with an extremely low detection limit of 162 fM. Also, the developed biosensor shows great reproducibility, acceptable stability, and reliable selectivity. Therefore, it offers a simple but effective aptasensor construction strategy for trace detection of DCF and is anticipated to show great potential for environmental applications.

Coating Persistent Luminescence Nanoparticles With Hydrophilic Polymers for in vivo Imaging.

Persistent luminescence nanoparticles (PLNPs) are innovative nanomaterials highly useful for bioimaging applications. Indeed, due to their particular optical properties, i.e., the ability to store the excitation energy before slowly releasing it for a prolonged period of time, they allow in vivo imaging without auto-fluorescence and with a high target to background ratio. However, as for most nanoparticles (NPs), without any special surface coating, they are rapidly opsonized and captured by the liver after systemic injection into small animals. To overcome this issue and prolong nanoparticle circulation in the bloodstream, a new stealth strategy was developed by covering their surface with poly(N-2-hydroxypropyl)methacrylamide (pHPMA), a highly hydrophilic polymer widely used in nanomedicine. Preliminary in vivo imaging results demonstrated the possibility of pHPMA as an alternative strategy to cover ZnGa2O4:Cr NPs to delay their capture by the liver, thereby providing a new perspective for the formulation of stealth NPs.

Degradation of ZnGa2O4:Cr3+ luminescent nanoparticles in lysosomal-like medium.

The ultimate goal of in vivo imaging is to provide safe tools to probe the inside of a body in order to obtain pathological information, monitor activities, and examine disease progression or regression. In this context zinc gallate doped with chromium III (ZGO) nanoparticles with persistent luminescence properties have been previously developed, and their biodistribution as well as in vitro toxicity were evaluated. However, to date, nothing is known about their potential transformations in biological media, which may hinder their biomedical applications. In order to know if these nanoparticles could degrade, the present work consists of studying their fate over time depending on both their coating and the aqueous media in which they are dispersed. ZGO nanoparticles have been dispersed in three different aqueous solutions for up to 90 days and characterized by numerous techniques. Among the evaluated dispersion media, Artificial Lysosomal Fluid (ALF) mimicking the intracellular lysosome environment elicited significant degradation of ZGO nanoparticles. The chelating agents present in ALF have proved to play a major role in the degradation of the ZGO, by stabilizing the nanoparticles and increasing the contact. An important time decrease of the luminescence properties has also been observed, which correlated with the release of ions from ZGO nanoparticles as well as their decreasing size. This information is valuable since it indicates, for the first time, the long-term degradation of persistent luminescent nanoprobes in an in vivo like model medium. Therefore, possible elimination of the imaging probes after in vivo preclinical applications could be foreseen.

Mucoadhesive thermosensitive hydrogel for the intra-tumoral delivery of immunomodulatory agents, in vivo evidence of adhesion by means of non-invasive imaging techniques.

Intratumoral injection of biocompatible gels is increasingly used for the sustained delivery of drugs and vaccines to enhance the anti-cancer immune response. Granulocyte-macrophage colony stimulating factor (GM-CSF) has become an attractive adjuvant thanks to its ability to boost the antitumor immune response by inducing proliferation, maturation and migration of the dendritic-cells (DCs) and the differentiation of lymphocytes. Killed Mycobacteria, such as Heat-killed Mycobacterium tuberculosis (HKMT) have been used in several studies as TLR-2 agonist to increase maturation of DCs. In this study, we designed a mucoadhesive thermosensitive formulation for the local delivery of GM-CSF and HKMT in order to enhance DCs activation and improve the local antitumor immune response. This formulation was selected based on its elastic and mucoadhesive properties obtained thanks to rheological studies. More importantly, intratumoral residence time of the labelled gel and protein were evidenced by means of MRI and non invasive in vivo optical imaging. Then, the efficacy of the combination of immunomodulators loaded thermogel was demonstated in vitro and in vivo. The selected thermogel exhibits rheological properties which confer a good elasticity and increased residence time of the immunostimulatory agents in the tumor, thus increasing the recruitment of DCs and T cytotoxic CD8+ lymphocytes.

Imaging and therapeutic applications of persistent luminescence nanomaterials.

The development of probes for biomolecular imaging and diagnostics is a very active research area. Among the different imaging modalities, optics emerged since it is a noninvasive and cheap imaging technique allowing real time imaging. In vitro, this technique is very useful however in vivo, fluorescence suffers from low signal-to-noise ratio due to tissue autofluorescence under constant excitation. To address this limitation, novel types of optical nanoprobes are actually being developed and among them, persistent luminescence nanoparticles (PLNPs), with long lasting near-infrared (NIR) luminescence capability, allows doing optical imaging without constant excitation and so without autofluorescence. This review will begin by introducing the physical phenomenon associated to the long luminescence decay of such nanoprobes, from minutes to hours after ceasing the excitation. Then we will show how this property can be used to develop in vivo imaging probes and also more recently nanotheranostic agents. Finally, preliminary data on their biocompatibility will be mentioned and we will conclude by envisioning on the future applications and improvements of such nanomaterials.

Novel Perfluorinated Triblock Amphiphilic Copolymers for Lipid-Shelled Microbubble Stabilization.

Amphiphilic triblock (Atri) copolymers made of perfluorinated alkyl chain linked to hydrocarbon chain and methoxy-poly(ethylene glycol) of three different molecular weights were synthesized. In vitro evaluation demonstrated that these new compounds were noncytotoxic. Characterization and interaction of each triblock copolymer with a branched polyamine myristoyl lipid (2-{3[bis-(3-amino-propyl)-amino]-propylamino}- N-ditetradecyl carbamoyl methyl-acetamide, DMAPAP) were studied by the Langmuir film method and thermal analysis. The triblock copolymer/cationic lipids (1:10, w/w) were mixed with perfluorobutane gas to form microbubbles (MBs). The latter were characterized by optical microscopy to get the microbubble size and concentration by densimetry to determine the amount of encapsulated gas and by ultrasound to assess oscillation properties. Atri with the lowest and intermediate weights were shown to interact with the cationic lipid DMAPAP and stabilize the Langmuir film. In that case, monodisperse microbubbles ranging from 2.3 ± 0.1 to 2.8 ± 0.1 µm were obtained. The proportion of encapsulated gas within the MB shell increased up to 3 times after the incorporation of the copolymer with the lowest and intermediate weights. Moreover, the acoustic response of the microbubbles was maintained in the presence of the copolymers.

Nanohybrids with Magnetic and Persistent Luminescence Properties for Cell Labeling, Tracking, In Vivo Real-Time Imaging, and Magnetic Vectorization.

Once injected into a living organism, cells diffuse or migrate around the initial injection point and become impossible to be visualized and tracked in vivo. The present work concerns the development of a new technique for therapeutic cell labeling and subsequent in vivo visualization and magnetic retention. It is hypothesized and subsequently demonstrated that nanohybrids made of persistent luminescence nanoparticles and ultrasmall superparamagnetic iron oxide nanoparticles incorporated into a silica matrix can be used as an effective nanoplatform to label therapeutic cells in a nontoxic way in order to dynamically track them in real-time in vitro and in living mice. As a proof-of-concept, it is shown that once injected, these labeled cells can be visualized and attracted in vivo using a magnet. This first step suggests that these nanohybrids represent efficient multifunctional nanoprobes for further imaging guided cell therapies development.

Gadolinium and Platinum in Tandem: Real-time Multi-Modal Monitoring of Drug Delivery by MRI and Fluorescence Imaging.

A novel dual-imaging cisplatin-carrying molecular cargo capable of performing simultaneous optical and MR imaging is reported herein. This long-lasting MRI contrast agent (r1 relaxivity of 23.4 mM-1s-1 at 3T, 25 oC) is a photo-activated cisplatin prodrug (PtGdL) which enables real-time monitoring of anti-cancer efficacy. PtGdL is a model for monitoring the drug delivery and anti-cancer efficacy by MRI with a much longer retention time (24 hours) in several organs such as renal cortex and spleen than GdDOTA and its motif control GdL. Upon complete release of cisplatin, all PtGdL is converted to GdL enabling subsequent MRI analyses of therapy efficacy within its reasonably short clearance time of 4 hours. There is also responsive fluorescence enhancement for monitoring by photon-excitation.

Electrokinetic Hummel-Dreyer characterization of nanoparticle-plasma protein corona: The non-specific interactions between PEG-modified persistent luminescence nanoparticles and albumin.

Nanoparticles (NPs) play an increasingly important role in the development of new biosensors, contrast agents for biomedical imaging and targeted therapy vectors thanks to their unique properties as well as their good detection sensitivity. However, a current challenge in developing such NPs is to ensure their biocompatibility, biodistribution, bioreactivity and in vivo stability. In the biomedical field, the adsorption of plasmatic proteins on the surface of NPs impacts on their circulation time in blood, degradation, biodistribution, accessibility, the efficiency of possible targeting agents on their surface, and their cellular uptake. NP surface passivation is therefore a very crucial challenge in biomedicine. We developed herein for the first time an electrokinetic Hummel-Dreyer method to quantitatively characterize the formation of protein corona on the surface of NPs. This strategy was designed and optimized to evaluate the non specific binding of bovine serum albumin with the recently discovered PEG-functionalized ZnGa1.995Cr0.005O4 persistent luminescence NPs developed for in vivo biological imaging. The binding strength and the number of binding sites were determined at different ionic strengths. This methodology opens the way to an easy, low sample- and low time-consuming evaluation of the impact of NP surface modification on protein-corona formation and therefore on their potential for various bio-medical applications.

Photo-stimulation of persistent luminescence nanoparticles enhances cancer cells death.

Persistent luminescence nanoparticles made of ZnGa1.995Cr0.005O4 (ZGO-NPs) are innovative nanomaterials that emit photons during long periods of time after the end of the excitation, allowing their use as diagnosis probes for in vivo optical imaging. During the excitation process, a part of the energy is stored in traps to further emit photons over long time. However, we observed in this study that some of the energy reduces molecular oxygen to produce reactive oxygen species (ROS). Following this observation, theoxidative stress induction and cytotoxic effects of these NPs were investigated on human breast cancer cells. The results indicate that ROS production was stimulated by exposition of the hydroxylated ZGO-NPs to UV or visible light, and the oxidative stress induced in cells after internalization can be directly correlated to their dose-dependent inhibition of cell viability. On the contrary, PEGylated ZGONPs were not uptaken by cells and have no effect on the production of ROS. Thus, the cell viability was not altered by these nanoparticles. This study reveals the importance of considering light irradiation and surface coating of luminescent nanoparticles toxicity which open new perspectives for their use in photodynamic therapy.