Small Warriors of Nature: Novel Red Emissive Chlorophyllin Carbon Dots Harnessing Fenton-Fueled Ferroptosis for In Vitro and In Vivo Cancer Treatment.

The appeal of carbon dots (CDs) has grown recently, due to their established biocompatibility, adjustable photoluminescence properties, and excellent water solubility. For the first time in the literature, copper chlorophyllin-based carbon dots (Chl-D CDs) are successfully synthesized. Chl-D CDs exhibit unique spectroscopic traits and are found to induce a Fenton-like reaction, augmenting photodynamic therapy (PDT) efficacies via ferroptotic and apoptotic pathways. To bolster the therapeutic impact of Chl-D CDs, a widely used cancer drug, temozolomide, is linked to their surface, yielding a synergistic effect with PDT and chemotherapy. Chl-D CDs' biocompatibility in immune cells and in vivo models showed great clinical potential.Proteomic analysis was conducted to understand Chl-D CDs' underlying cancer treatment mechanism. The study underscores the role of reactive oxygen species formation and pointed toward various oxidative stress modulators like aldolase A (ALDOA), aldolase C (ALDOC), aldehyde dehydrogenase 1B1 (ALDH1B1), transaldolase 1 (TALDO1), and transketolase (TKT), offering a deeper understanding of the Chl-D CDs' anticancer activity. Notably, the Chl-D CDs' capacity to trigger a Fenton-like reaction leads to enhanced PDT efficiencies through ferroptotic and apoptotic pathways. Hence, it is firmly believed that the inherent attributes of Chl-CDs can lead to a secure and efficient combined cancer therapy.

DFMO Carbon Dots for Treatment of Neuroblastoma and Bioimaging.

Neuroblastoma (NB) is a pediatric malignancy affecting the peripheral nervous system. Despite recent advancements in treatment, many children affected with NB continue to submit to this illness, and new therapeutic strategies are desperately needed. In recent years, studies of carbon dots (CDs) as nanocarriers have mostly focused on the delivery of anticancer agents because of their biocompatibility, good aqueous dissolution, and photostability. Their fluorescence properties, surface functionalities, and surface charges differ on the basis of the type of precursors used and the synthetic approach implemented. At present, most CDs are used as nanocarriers by directly linking them either covalently or electrostatically to drug molecules. Though most modern CDs are synthesized from large carbon macromolecules and conjugated to anticancerous drugs, constructing CDs from the anticancerous drugs and precursors themselves to increase antitumoral activity requires further investigation. Herein, CDs were synthesized using difluoromethylornithine (DFMO), an irreversible ornithine decarboxylase inhibitor commonly used in high-risk neuroblastoma treatment regiments. In this study, NB cell lines, SMS-KCNR and SK-N-AS, were treated with DFMO, the newly synthesized DFMO CDs, and conventional DFMO conjugated to black carbon dots. Bioimaging was done to determine the cellular localization of a fluorescent drug over time. The mobility of DNA mixed with DFMO CDs was evaluated by gel electrophoresis. DFMO CDs were effectively synthesized from DFMO precursor and characterized using spectroscopic methods. The DFMO CDs effectively reduced cell viability with increasing dose. The effects were dramatic in the N-MYC-amplified line SMS-KCNR at 500 µM, which is comparable to high doses of conventional DFMO at a 60-fold lower concentration. In vitro bioimaging as well as DNA electrophoresis showed that synthesized DFMO CDs were able to enter the nucleus of neuroblastoma cells and neuronal cells and interact with DNA. Our new DFMO CDs exhibit a robust advantage over conventional DFMO because they induce comparable reductions in viability at a dramatically lower concentration.

Structure-Activity Relationship of Carbon Nitride Dots in Inhibiting Tau Aggregation.

Due to the numerous failed clinical trials of anti-amyloid drugs, microtubule associated protein tau (MAPT) now stands out as one of the most promising targets for AD therapy. In this study, we report for the first time the structure-dependent MAPT aggregation inhibition of carbon nitride dots (CNDs). CNDs have exhibited great promise as a potential treatment of Alzheimer's disease (AD) by inhibiting the aggregation of MAPT. In order to elucidate its structure-activity relationship, CNDs were separated via column chromatography and five fractions with different structures were obtained that were characterized by multiple spectroscopy methods. The increase of surface hydrophilic functional groups is consistent with the increase of polarity from fraction 1 to 5. Particle sizes (1-2 nm) and zeta potentials (~-20 mV) are similar among five fractions. With the increase of polarity from fraction 1 to 5, their MAPT aggregation inhibition capacity was weakened. This suggests hydrophobic interactions between CNDs and MAPT, validated via molecular dynamics simulations. With a zebrafish blood-brain barrier (BBB) model, CNDs were observed to cross the BBB through passive diffusion. CNDs were also found to inhibit the generation of multiple reactive oxygen species, which is an important contributor to AD pathogenesis.

Drug delivery of memantine with carbon dots for Alzheimer's disease: blood-brain barrier penetration and inhibition of tau aggregation.

Neurofibrillary tangle, composed of aggregated tau protein, is a pathological hallmark of Alzheimer's disease (AD). The inhibition of tau aggregation is therefore an important direction for AD drug discovery. In this work, we explored the efficacy of two types of carbon dots in targeting tau aggregation, as versatile nano-carriers for the development of carbon dots (CDs)-based AD therapy. We carried out synthesis, biophysical and biochemical characterizations of two types of CDs, namely, carbon nitride dots (CNDs) and black carbon dots (B-CDs). CDs, which are biocompatible and non-toxic, were successfully conjugated with memantine hydrochloride (MH) through EDC/NHS mediated amidation reactions followed by systematic characterizations using various biophysical techniques including UV-vis spectroscopy (UV-vis), photoluminescence (PL), Fourier-transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), mass spectrometry (MS), Transmission electron microscopy (TEM) and atomic force microscopy (AFM). The surface diversity along with small particle sizes of CDs allowed facile delivery of MH across the blood-brain barrier (BBB), as demonstrated using a zebrafish in vivo model. The tau aggregation inhibition experiments were conducted using the thioflavin-T (ThT) assay to identify the most effective inhibitor. The kinetics and magnitude of tau aggregation were measured in the presence of CDs, which demonstrates that both B-CDs-MH and B-CDs alone are the most effective inhibitors of tau aggregation with IC50 values of 1.5 ± 0.3 and 1.6 ± 1.5 µg/mL, respectively. Taken together, our findings hold therapeutic significance to enhance the efficient delivery of MH to target AD pathology in the brain for improved efficacy.

Rose Bengal and Riboflavin Mediated Photodynamic Antimicrobial Therapy Against Selected South Florida Nocardia Keratitis Isolates.

Purpose: To examine and compare the efficacy of in vitro growth inhibition using rose bengal and riboflavin photodynamic antimicrobial therapy (PDAT) for Nocardia keratitis isolates. Methods: Nocardia asteroides complex, Nocardia amikacinitolerans, and Nocardia farcinica species were isolated from patients with confirmed Nocardia keratitis. Isolates were tested against three experimental groups: (1) no photosensitizer/no irradiation, (2) photosensitizer/no irradiation, and (3) photosensitizer/irradiation. Each isolate was prepared in suspension to a concentration of 1.5 × 108 CFU/mL. Bacterial suspensions were mixed with water or prepared 0.1% photosensitizer solution for a final bacterial concentration of 1.5 × 107 CFU/mL. Aliquots of 1 mL were plated on 5% sheep blood agar. Rose bengal and riboflavin PDAT plates were irradiated for 15 minutes with a 525- or 375-nm custom 6-mW/cm2 powered light source for a total fluence of 5.4 J/cm2. All experimental groups were repeated in triplicate. Plates were incubated in a 35°C non-CO2 incubator for 96 hours and photographed. Percent inhibition was evaluated using LabVIEW-based software. Results: All strains of Nocardia tested with 0.1% rose bengal and irradiated for 15 minutes demonstrated statistically significant inhibition of growth (P < 0.05). No other experimental groups displayed any bacterial inhibition. Conclusions: Rose bengal is superior to riboflavin PDAT against selected Nocardia isolates. In vivo testing is warranted to investigate the utility of rose bengal PDAT for severe Nocardia keratitis. Translational Relevance: In vitro results for three clinical strains of Nocardia support the possible use of rose bengal PDAT as a complementary treatment of Nocardia keratitis.

Development of Red-Emissive Carbon Dots for Bioimaging through a Building Block Approach: Fundamental and Applied Studies.

In recent years, many researchers have struggled to obtain carbon dots (CDs) that possess strong photoluminescence in the red region of light. Success in this area has been limited, although the past few years have brought several promising reports on this topic. The most successful efforts in this area still seem to struggle from a lack of dispersibility/reduced emission in water. This work endeavors to understand the formation process of CDs that do not possess strong performance in an aqueous environment and to improve their capabilities in bioimaging. o-Phenylenediamine (o-PDA) is used along with various precursors in several different solvents (varying acidic and oxidative strengths) to understand the formation process behind the structure leading to red emission that is sensitive to water. These results showed that the combination of acid properties and oxidation is essential for this process, and the important reactions are oligomerization of o-PDA and the crosslinking of these oligomers to form aromatic structural segments of CDs. These CDs are shown to be capable of quantitatively detecting water in organic solvents. Additionally, we have shown that conjugation with transferrin remarkably enhances the biocompatibility of these CDs. Transferrin-conjugated CDs with better biocompatibility were applied to bioimaging studies of neuroblastoma cell lines with N-myc and non-N-myc gene amplification, for the first time. Furthermore, CDs showed versatile bioimaging capability toward a highly aggressive neuroblastoma subgroup of tumors. The importance of creating red-emissive CDs has been well established, and this work is an important step toward understanding their formation and realizing their use in biological systems.

The use of nanotechnology to combat liver cancer: Progress and perspectives.

Liver cancer is one of the most common cancers worldwide and is also one of the most difficult cancers to treat, resulting in almost one million deaths per year, and the danger of this cancer is compounded when the tumor is nonresectable. Hepatocellular carcinoma (HCC) is the most common type of liver cancer and has the third highest mortality rate worldwide. Considering the morbid statistics surrounding this cancer it is a popular research topic to target for better therapy practices. This review summarizes the role of nanotechnology in these endeavors. Nanoparticles (NPs) are a very broad class of material and many different kinds have been used to potentially combat liver cancer. Gold, silver, platinum, metal oxide, calcium, and selenium NPs as well as less common materials are all inorganic NPs that have been used as a therapeutic, carrier, or imaging agent in drug delivery systems (DDS) and these efforts are described. Carbon-based NPs, including polymeric, polysaccharide, and lipid NPs as well as carbon dots, have also been widely studied for this purpose and the role they play in DDS for the treatment of liver cancer is illustrated in this review. The multifunctional nature of many NPs described herein, allows these systems to display high anticancer activity in vitro and in vivo and highlights the advantage of and need for combinatorial therapy in treating this difficult cancer. These works are summarized, and future directions are presented for this promising field.

Carbon Dots: A Future Blood-Brain Barrier Penetrating Nanomedicine and Drug Nanocarrier.

Drug delivery across the blood-brain barrier (BBB) is one of the biggest challenges in modern medicine due to the BBB's highly semipermeable property that limits most therapeutic agents of brain diseases to enter the central nervous system (CNS). In recent years, nanoparticles, especially carbon dots (CDs), exhibit many unprecedented applications for drug delivery. Several types of CDs and CD-ligand conjugates have been reported successfully penetrating the BBB, which shows a promising progress in the application of CD-based drug delivery system (DDS) for the treatment of CNS diseases. In this review, our discussion of CDs includes their classification, preparations, structures, properties, and applications for the treatment of neurodegenerative diseases, especially Alzheimer's disease (AD) and brain tumor. Moreover, abundant functional groups on the surface, especially amine and carboxyl groups, allow CDs to conjugate with diverse drugs as versatile drug nanocarriers. In addition, structure of the BBB is briefly described, and mechanisms for transporting various molecules across the BBB and other biological barriers are elucidated. Most importantly, recent developments in drug delivery with CDs as BBB-penetrating nanodrugs and drug nanocarriers to target CNS diseases especially Alzheimer's disease and brain tumor are summarized. Eventually, future prospects of the CD-based DDS are discussed in combination with the development of artificial intelligence and nanorobots.

Surface Chemistry Studies on the Formation of Mixed Stearic Acid/Phenylalanine Dehydrogenase Langmuir and Langmuir-Blodgett Films.

This work investigates the physicochemical properties of mixed stearic acid (HSt)/phenylalanine dehydrogenase enzyme (PheDH) Langmuir films and their immobilization onto solid supports as Langmuir-Blodgett (LB) films. PheDH from the aqueous subphase enters the surfactant matrix up to an exclusion surface pressure of 25.3 mN/m, leading to the formation of stable and highly condensed mixed Langmuir monolayers. Hydrophobic interactions between the enzyme and HSt nonpolar groups tuned the secondary structure of PheDH, evidenced by the presence of ß-sheet structures as demonstrated by infrared and circular dichroism spectra. The floating monolayers were successfully transferred to solid quartz supports, yielding Y-type LB films, and then characterized employing fluorescence, circular dichroism, and microscopic techniques, which indicated that PheDH was co-immobilized with HSt proportionally to the number of transferred layers. The enzyme fluidized the HSt monolayers, reducing their maximum dipoles when condensed to their maximum, and disorganized the alkyl chains of the fatty acid, as detected with infrared spectroscopy. The stability of the mixed floating monolayers enabled their transfer to solid supports as LB films, which is important for producing optical and electrochemical sensors for phenylalanine whose molecular architecture can be controlled with precision.

Rose Bengal Photodynamic Antimicrobial Therapy: A Pilot Safety Study.

PURPOSE: To evaluate the in vivo corneal changes after Rose Bengal photodynamic antimicrobial therapy (RB-PDAT) treatment in New Zealand White rabbits. METHODS: Sixteen rabbits were divided into 5 groups. All groups underwent deepithelialization of an 8 mm diameter area in the central cornea. Group 1: balanced salt solution drops only, group 2: 0.2% RB only, group 3: green light exposure (525 nm, 5.4 J/cm2) only, group 4: 0.1% RB-PDAT, and group 5: 0.1% RB-PDAT. All rabbits were followed clinically. Group 5 rabbits were followed using anterior segment optical coherence tomography (AS-OCT) and clinically. On day 35 after initial treatment, 1 rabbit from group 5 was re-exposed to green light (5.4 J/cm2) to evaluate reactivation of the remaining RB dye, and terminal deoxynucleotyl transferase-mediated UTP-biotin-nick-end labeling assay was performed on corneal cryosections. RESULTS: Complete reepithelization was observed, and corneas remained clear after treatment in all groups. In group 5, AS-OCT revealed a cross-linking demarcation line. AS-OCT showed RB fluorescence and collagen cross-linking in all treated eyes of group 5 animals after 5 weeks of treatment. Photobleached RB retention in the corneal stroma was corroborated by fluorescence confocal microscopy on frozen sections. There was no evidence of a sustained cytotoxic effect through terminal deoxynucleotyl transferase-mediated UTP-biotin-nick-end labeling at 5 weeks. CONCLUSIONS: RB-PDAT with 0.1% RB is a safe procedure. There was no difference clinically and on histopathology compared with control groups. In eyes where RB dye is retained in the corneal stroma after 1 month of treatment, oxidative stress is not evidenced at long term.

Gel-like carbon dots: A high-performance future photocatalyst.

To protect water resources, halt waterborne diseases, and prevent future water crises, photocatalytic degradation of water pollutants arouse worldwide interest. However, considering the low degradation efficiency and risk of secondary pollution displayed by most metal-based photocatalysts, highly efficient and environmentally friendly photocatalysts with appropriate band gap, such as carbon dots (CDs), are in urgent demand. In this study, the photocatalytic activity of gel-like CDs (G-CDs) was studied using diverse water pollution models for photocatalytic degradation. The degradation rate constants demonstrated a remarkably enhanced photocatalytic activity of G-CDs compared with most known CD species and comparability to graphitic carbon nitride (g-C3N4). In addition, the rate constant was further improved by 1.4 times through the embedment of g-C3N4 in G-CDs to obtain CD-C3N4. Significantly, the rate constant was also higher than that of g-C3N4 alone, revealing a synergistic effect. Moreover, the use of diverse radical scavengers suggested that the main contributors to the photocatalytic degradation with G-CDs alone were superoxide radicals (O2-) and holes that were, however, substituted by O2- and hydroxyl radicals (OH) due to the addition of g-C3N4. Furthermore, the photocatalytic stabilities of G-CDs and CD-C3N4 turned out to be excellent after four cycles of dye degradation were performed continuously. Eventually, the nontoxicity and environmental friendliness of G-CDs and CD-C3N4 were displayed with sea urchin cytotoxicity tests. Hence, through various characterizations, photocatalytic degradation and cytotoxicity tests, G-CDs proved to be an environmentally friendly and highly efficient future photocatalyst.

Metformin derived carbon dots: Highly biocompatible fluorescent nanomaterials as mitochondrial targeting and blood-brain barrier penetrating biomarkers.

Carbon dots (CDs) have been intensively studied since their discovery in 2004 because of their unique properties such as low toxicity, excellent biocompatibility, high photoluminescence (PL) and good water dispersibility. In this study metformin derived carbon dots (Met-CDs) were synthesized using a microwave assisted method. Met-CDs were meticulously characterized using ultra-violet spectroscopy (UV-vis), photoluminescence (PL), Fourier Transform Infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), atomic force (AFM) and transmission electron (TEM) microscopies. According to results of cytotoxicity studies, Met-CDs possess low-toxicity and excellent biocompatibility towards both non-tumor and tumor cell lines indicating that Met-CDs are outstanding candidates for living cell bioimaging studies. Furthermore, bioimaging studies have displayed that Met-CDs can penetrate the cell membrane and disperse throughout the cell structure including the nucleus and mitochondria. More specifically, Met-CDs tend to start localizing selectively inside the mitochondria of cancer cells, but not of non-tumor cells after 1 h of incubation. Finally, a zebrafish study confirmed that Met-CDs cross the blood-brain barrier (BBB) without the need of any other ligands. In summary, this study presents synthesis of Met-CDs which feature abilities such as mitochondrial and nucleus localizations along with BBB penetration.

Detection of singlet oxygen luminescence for experimental corneal rose bengal photodynamic antimicrobial therapy.

Rose bengal photodynamic antimicrobial therapy (RB-PDAT) treats corneal infection by activating rose bengal (RB) with green light to produce singlet oxygen (1O2). Singlet oxygen dosimetry can help optimize treatment parameters. We present a 1O2 dosimeter for detection of 1O2 generated during experimental RB-PDAT. The system uses a 520 nm laser and an InGaAs photoreceiver with bandpass filters to detect 1O2 luminescence during irradiation. The system was validated in RB solutions and ex vivo in human donor eyes. The results demonstrate the feasibility of 1O2 dosimetry in an experimental model of RB-PDAT in the cornea.

Facile Synthesis of "Boron-Doped" Carbon Dots and Their Application in Visible-Light-Driven Photocatalytic Degradation of Organic Dyes.

Carbon dots (C-dots) were facilely fabricated via a hydrothermal method and fully characterized. Our study shows that the as-synthesized C-dots are nontoxic, negatively charged spherical particles (average diameter 4.7 nm) with excellent water dispersion ability. Furthermore, the C-dots have a rich presence of surface functionalities such as hydroxyls and carboxyls as well as amines. The significance of the C-dots as highly efficient photocatalysts for rhodamine B (RhB) and methylene blue (MB) degradation was explored. The C-dots demonstrate excellent photocatalytic activity, achieving 100% of RhB and MB degradation within 170 min. The degradation rate constants for RhB and MB were 1.8 × 10-2 and 2.4 × 10-2 min-1, respectively. The photocatalytic degradation performances of the C-dots are comparable to those metal-based photocatalysts and generally better than previously reported C-dots photocatalysts. Collectively considering the excellent photocatalytic activity toward organic dye degradation, as well as the fact that they are facilely synthesized with no need of further doping, compositing, and tedious purification and separation, the C-dots fabricated in this work are demonstrated to be a promising alternative for pollutant degradation and environment protection.

Direct conjugation of distinct carbon dots as Lego-like building blocks for the assembly of versatile drug nanocarriers.

As a promising drug nanocarrier, carbon dots (CDs) have exhibited many excellent properties. However, some properties such as bone targeting and crossing the blood-brain barrier (BBB) only apply to a certain CD preparation with limited drug loading capacity. Therefore, it is significant to conjugate distinct CDs to centralize many unique properties on the novel drug nanocarrier. Considering that CDs have abundant and tunable surface functionalities, in this study, a direct conjugation was initiated between two distinct CD models, black CDs (B-CDs) and gel-like CDs (G-CDs) via an amidation reaction. As a result of conjugation at a mass ratio of 5:3 (B-CDs to G-CDs) and a two-step purification process, the conjugate, black-gel CDs (B-G CDs) (5:3) inherited functionalities from both CDs and obtained an enhanced thermostability, aqueous stability, red-shifted photoluminescence (PL) emission, and a figure-eight shape with a width and length of 3 and 6 nm, respectively. In addition, the necessity of high surface primary amine (NH2) content in the CD conjugation was highlighted by replacing G-CDs with other CDs with lower surface NH2 content. Meanwhile, the carboxyl groups (COOH) on G-CDs were not enough to trigger self-conjugation between G-CDs. Moreover, the drug loading capacity was enhanced by 54.5% from B-CDs to B-G CDs (5:3). Furthermore, when the mass ratio of B-CDs to G-CDs was decreased from 5:30, 5:100 to 5:300, the obtained nanostructures revealed a great potential of CDs as Lego-like building blocks. Also, bioimaging of zebrafish demonstrated that various B-G CDs exhibited properties of both bone targeting and crossing the BBB, which are specific properties of B-CDs and G-CDs, respectively.

Tyrosinase enzyme Langmuir monolayer: Surface chemistry and spectroscopic study.

This study investigates the surface chemistry properties of the tyrosinase enzyme Langmuir monolayer at air-aqueous interface using sodium chloride in the subphase to induce the surface activity of the enzyme. Investigation of surface packing and stability of the tyrosinase Langmuir monolayer were performed using surface chemistry experiments while spectroscopic analysis was done to study enzyme conformation. It was found that the tyrosinase enzyme forms a fluid film at air-aqueous interface with good stability as shown by compression-decompression cycles experiments and stability measurements at various surface pressures. UV-vis absorption and fluorescence measurements at different surface pressures revealed that the Langmuir monolayer has good homogeneity with no evidence of aggregates during compression. To gain insight on the conformation of tyrosinase Langmuir monolayer p-polarized infrared-reflection absorption spectroscopy was used. It was found that at high surface pressures the predominant secondary structures were ß-sheets while at lower surface pressure both α -helices and ß-sheets were present. The circular dichroism spectra were obtained by transferring the Langmuir monolayer at 10 mN.m-1 to a solid quartz support (Langmuir-Blodgett film, LB film), which showed that the major conformation present were α-helices. Images from the immobilized LB films were obtained using atomic force microscopy which showed homogenous and regular deposition with a mean thickness ranging from 3 to 4 nm.

Surface chemistry and spectroscopic studies of the native phenylalanine dehydrogenase Langmuir monolayer at the air/aqueous NaCl interface.

This study investigates the main aspects of the surface behavior of the native phenylalanine dehydrogenase (PheDH) enzyme at the air/aqueous interface employing a saline subphase to induce the enzyme surface activity. Surface chemistry experiments were performed in order to determine the surface packing and stability of the formed layer, while spectroscopic experiments provided information regarding its secondary structure conformation. It was found that the PheDH enzyme forms a fluid film, which is quite homogeneous throughout its entire compression, being stable for long periods of time with no significant evidence of aggregates or irreversible domains during interfacial compression/decompression processes. The main secondary structures of the interfacial PheDH film were accessed via in situ reflectance-absorbance infrared spectroscopy, indicating a majority presence of α-helices, which were maintained after the film transfer to solid muscovite supports. The immobilized films presented a homogeneous and regular deposition, with controlled roughness and a mean thickness in the range of 8-10 nm.

Carbon Dots: Diverse Preparation, Application, and Perspective in Surface Chemistry.

Carbon dots (CDs) are a novel class of nanoparticles with excellent properties. The development of CDs involves versatile synthesis, characterization, and various applications. However, the importance of surface chemistry of CDs, especially in applications, is often underestimated. In fact, the study of the surface chemistry of CDs is of great significance in the explanation of the unique properties of CDs as well as the pursuit of potential applications. In this feature article, we do not only introduce the development of CDs in our group but also highlight their applications where surface chemistry plays a critical role.

Close-Packed Langmuir Monolayers of Saccharide-Based Carbon Dots at the Air-Subphase Interface.

Carbon dots (CDs) are zero-dimensional carbon-based spherical nanoparticles with diameters less than 10 nm. Here, we report for the first time CDs forming stable Langmuir monolayers at the air-subphase interface. Langmuir monolayers are of great interest both fundamentally to study the interactions at the interfaces and for many applications such as the development of sensors. However, CDs usually do not form Langmuir monolayers because of their highly hydrophilic nature. In this study, amphiphilic CDs were prepared through hydrothermal carbonization using saccharides as the precursors. The surface chemistry behavior and optical properties of CDs at the air-subphase interface were studied. CDs derived from saccharides consistently formed stable Langmuir monolayers which show all essential phases, namely, gas, liquid-expanded, liquid-condensed, and solid phases. The compression-decompression cycle method showed minimum hysteresis (4.3%), confirming the retaining capacity of the CDs as a monolayer. Limiting CD areas from surface pressure-area isotherm at the air-subphase interface were used to calculate the average diameter of the CDs at the air-subphase interface. UV/vis absorption spectra of CDs dispersed in water and in Langmuir monolayers had the same bands in the UV region. The intensity of the UV/vis absorption increases with increasing surface pressure at the air-subphase interface. Interestingly, photoluminescence (PL) of the Langmuir monolayer of CDs was excitation-independent, whereas the same CDs had excitation-dependent PL when dispersed in water.

Triple conjugated carbon dots as a nano-drug delivery model for glioblastoma brain tumors.

Most of the dual nano drug delivery systems fail to enter malignant brain tumors due to a lack of proper targeting systems and the size increase of the nanoparticles after drug conjugation. Therefore, a triple conjugated system was developed with carbon dots (C-dots), which have an average particle size of 1.5-1.7 nm. C-dots were conjugated with transferrin (the targeted ligand) and two anti-cancer drugs, epirubicin and temozolomide, to build the triple conjugated system in which the average particle size was increased only up to 3.5 nm. In vitro studies were performed with glioblastoma brain tumor cell lines SJGBM2, CHLA266, CHLA200 (pediatric) and U87 (adult). The efficacy of the triple conjugated system (dual drug conjugation along with transferrin) was compared to those of dual conjugated systems (single drug conjugation along with transferrin), non-transferrin C-dots-drugs, and free drug combinations. Transferrin conjugated samples displayed the lowest cell viability even at a lower concentration. Among the transferrin conjugated samples, the triple conjugated system (C-dots-trans-temo-epi (C-DT)) was more strongly cytotoxic to brain tumor cell lines than dual conjugated systems (C-dots-trans-temo (C-TT) and C-dots-trans-epi (C-ET)). C-DT increased the cytotoxicity to 86% in SJGBM2 at 0.01 µM while C-ET and C-TT reduced it only to 33 and 8%, respectively. Not only did triple conjugated C-DT increase the cytotoxicity, but also the two-drug combination in C-DT displayed a synergistic effect.