Cellular Uptake Studies Research Articles

Scalable Manufacturing Method for Model Protein-Loaded PLGA Nanoparticles: Biocompatibility, Trafficking and Release Properties

Background and Objectives: Drug delivery systems (DDSs) offer efficient treatment solutions to challenging diseases such as central nervous system (CNS) diseases by bypassing biological barriers such as the blood–brain barrier (BBB). Among DDSs, polymeric nanoparticles (NPs), particularly poly(lactic-co-glycolic acid) (PLGA) NPs, hold an outstanding position due to their biocompatible and biodegradable qualities. Despite their potential, the translation of PLGA NPs from laboratory-scale production to clinical applications remains a significant challenge. This study aims to address these limitations by developing scalable PLGA NPs and evaluating their potential biological applications. Methods: We prepared blank and model-protein-loaded (albumin–FITC and wheat germ agglutinin-488 (WGA-488)) fluorescent PLGA NPs using the traditional double-emulsion method combined with the micro-spray-reactor system, a novel approach that enables fine particle production enabling scale-up applications. We tested the biocompatibility of the NPs in living RPMI 2650 and neuroblastoma cell lines, as well as their trafficking and uptake. Release kinetics of the encapsulated proteins were investigated through confocal microscopy and in vitro release studies, providing insights into the stability and functionality of the released proteins. Results: The formulation demonstrated sustained and prolonged protein release profiles. Importantly, cellular uptake studies revealed that the NPs were not internalized. Furthermore, encapsulated WGA-488 protein retained its functional activity after release, validating the integrity of the encapsulation and release processes. Conclusions: The proof-of-concept study on NP manufacturing and an innovative drug trafficking and release approach can bring new perspectives on scalable preparations of PLGA NPs and their biological applications.

Modulation of Donor in Purely Organic Triplet Harvesting AIE-TADF Photosensitizer for Image-guided Photodynamic Therapy.

Image-guided photodynamic therapy is acknowledged as one of the most demonstrative therapeutic modalities for cancer treatment because of its high precision, non-invasiveness, and improved imaging ability. A series of purely organic photosensitizers denoted as BTMCz, BTMPTZ, and BTMPXZ, have been designed and synthesized and are found to exhibit both thermally activated delayed fluorescence and aggregation-induced emission simultaneously. Experimental and theoretical studies are combined to reveal that modulation of the donor of the photosensitizer enables distinct thermally activated delayed fluorescence via a second-order spin-orbit perturbation mechanism involving lowest singlet charge-transfer and higher-lying triplet locally excited states, respectively. Further, different donor strengths and unique aggregations (H-, J- and X-type packings) greatly influence their color-tunable up-converted luminescence and endow them with superb dispersibility in water. The confocal microscopy-based cellular uptake study confirms the successful internalization of the nano-probes, while BTMCz enables the generation of reactive oxygen species (singlet oxygen) under white-light irradiation, enabling the efficient killing of cancer cells.

PEGylated solid lipid nanoparticles for the intranasal delivery of combination antiretroviral therapy composed of Atazanavir and Elvitegravir to treat NeuroAIDS.

Intranasal drug administration offers a promising strategy for delivering combination antiretroviral therapy (cART) directly to the central nervous system to treat NeuroAIDS, leveraging the nose-to-brain route to bypass the blood-brain barrier. However, challenges such as enzymatic degradation in the nasal mucosa, low permeability, and mucociliary clearance within the nasal cavity must first be addressed to make this route feasible. To overcome these barriers, this study developed solid lipid nanoparticles (SLNs) with varying PEGylation levels (0%, 5%, 10%, and 15% w/w of PEGylated lipid), co-encapsulated with Elvitegravir (EVG) and Atazanavir (ATZ) as an integrase and protease inhibitor, respectively. Pre-formulation studies confirmed the compatibility of the drugs with the excipients. Characterization showed that PEGylation reduces SLN size by approximately up to 12% while maintaining monodispersity and a high encapsulation efficiency of over 99% for both EVG and ATZ in their amorphous forms. Incubation of the formulations in artificial nasal mucus revealed that increased PEGylation consistently reduces nanoparticle aggregation and mean aggregate size, suggesting improved SLN stability in the mucus. Importantly, higher PEGylation levels significantly enhanced model drug permeability across the nasal mucus barrier by up to 10-fold. Lastly, cellular uptake studies using the RPMI 2650 nasal epithelial cell line indicated that PEGylation does not reduce nanoparticle uptake rates. These findings highlight the potential of PEGylated SLNs as an effective vehicle for enhancing the intranasal delivery of cART to treat NeuroAIDS. However, further in vivo studies are needed to confirm the brain targeting potential of this formulation.

Enhanced luminescence and stability of TFMDSA nanoparticles via polymer-induced aggregation for bioimaging.

In recent years, fluorescence imaging has occupied a very important position in the life science and biomedical fields. However, achieving nanomaterials for bioimaging with both high fluorescence quantum efficiency and high stability remains a significant challenge. Herein, we synthesized mPEG5K-PCL10K@TFMDSA and mPEG5K-PLLA10K@TFMDSA nanoparticles using polymer-induced aggregation. This method significantly enhanced the luminescence efficiency of TFMDSA nanoparticles in solution, attributed to improved intermolecular interactions and restricted molecular vibrations. The resulting nanoparticles exhibited exceptional optical stability over a period of seven days and demonstrated low cytotoxicity towards HeLa cells, making them highly suitable for bioimaging applications. Cellular uptake studies indicated that these nanoparticles were more efficiently internalized by HeLa cells compared to their amorphous counterparts, likely due to their unique square morphology. Our findings highlight the potential of polymer-induced aggregation in enhancing the optical properties and stability of TFMDSA nanomaterials, suggesting their promise as biofluorescent probes for cancer diagnosis and other biomedical applications.

Hyaluronic Acid-Based Hybrid Nanoparticles as Promising Carriers for the Intranasal Administration of Dimethyl Fumarate.

Dimethyl fumarate (DMF), the first-line oral therapy for relapsing-remitting multiple sclerosis, is rapidly metabolized into monomethyl fumarate. The DMF oral administration provokes gastrointestinal discomfort causing treatment withdrawal. The present study aimed to develop an innovative formulation for DMF nasal administration. Lipid-polymer hybrid nanoparticles (LPNs) were developed to improve DMF stability, limiting gastrointestinal side effects and increasing brain bioavailability by nose-to-brain targeting application. DMF-loaded and unloaded LPNs with or without hyaluronic acid (HA) were prepared using the nanoprecipitation via magnetic/mechanical stirring technique. Particle morphology and surface properties were evaluated; drug content, viscosity, and mucoadhesion were determined. Physico-chemical stability of LPNs and DMF in the LPNs was also explored. In vitro DMF permeation experiments were performed utilizing the PermeaPad®. The cytotoxicity and cellular uptake studies were performed using RPMI 2650 and SK-N-BE2 cell lines. DMF nose-to-brain delivery was evaluated by intranasally administering DMF-loaded LPNs to rats. LPNs with average sizes of 120-250 nm and a negative zeta potential -17.3 to -43 mV were obtained, primarily influenced by the presence of HA. HA assured drug stability up to 60 days and promoting the in vitro permeation of DMF compared to the free-DMF. HA greatly improved the viscosity and mucoadhesive properties. LPNs with and without HA did not exhibit any cytotoxicity and showed a rapid cell uptake starting from 15min to 2h with a best internalization after 1h of treatment in both epithelial and neuronal cell lines. Nasal administration of DMF-loaded LPNs allowed to quantify up to about 12μg/mL of DMF in the rat cerebrospinal fluid. The results highlight the role of HA in improving LPNs properties and performance as carrier of DMF for nasal administration. In particular, LPNs appear able to enter neurons and monolayers of epithelial cells, allowing to promote the nose-to-brain DMF delivery.

Impact of the hydrophilic-lipophilic balance of free-base and Zn(II) tricationic pyridiniumporphyrins and irradiation wavelength in PDT against the melanoma cell lines.

The amphiphilic and asymmetric structure of porphyrins, when used as photosensitizers (PSs) for photodynamic therapy (PDT), has been shown through numerous previous studies to be a very important property that facilitates their entry into cells, which improves their efficiency in PDT. In this work, two groups of cationic AB3 pyridiniumporphyrins, free-base and chelated with Zn(II), both substituted with alkyl chains of various lengths, were studied in PDT on melanoma cell lines. The aim was to investigate the impact of hydrophilic-lipophilic balance and Zn(II) chelation, and the importance of matching the irradiation wavelength to the optical properties of the PS on in vitro PDT efficiency. Therefore, spectroscopic studies, singlet oxygen production and lipophilicity as well as cellular uptake, localization and cytotoxicity studies of the two series of porphyrins were performed. In both series of porphyrins, the longest alkyl chain (17C-atoms long) enables the greatest internalization of the PS. Chelation with Zn(II) resulted in better physicochemical properties, but slower cellular internalization. As expected, free-base porphyrins were more PDT efficient than their Zn(II) complexes after 30-min photoactivation by low-fluence (2mW/cm2) red light (643nm). However the use of orange light (606nm) with the same fluence rate was more suitable for Zn(II) porphyrins and resulted in similar overall toxicity to their free-base analogues with similar lipophilicity. Although the highest phototoxicity was achieved with the PSs carrying the longest alkyl chain, TMPyP3-C13H27 and Zn(II)-TMPyP3-C13H27 proved to be the most promising candidates for use in PDT as they exhibit high phototoxicity, but also greater selectivity towards melanoma cell lines (MeWo and A375) compared to fibroblasts (HDF).

Engineering of redox-triggered polymeric lipid hybrid nanocarriers for selective drug delivery to cancer cells.

Tunable redox-sensitive polymeric-lipid hybrid nanocarriers (RS-PLHNCs) were fabricated using homogenization and nanoprecipitation methods. These nanocarriers were composed of novel redox-cholesterol with disulfide linkages and synthesized by conjugating cholesterol with dithiodipropionic acid via esterification. Berberine (BBR) was loaded into the fabricated nanocarriers to investigate the selective uptake of BBR by cancer cells as well as its release and enhanced cytotoxicity. The optimized BBR nanocarriers BBR NP-17 and -18 exhibited a spherical shape and uniform distribution, with a particle size of 124.7 ± 1.2 nm and 185.2 ± 1.6 nm and a zeta potential of -5.9 ± 2.5 mV and -20.3 ± 1.1 mV, respectively. These NCs released >80% BBR in a simulated intracellular tumor microenvironment (TME), while only 30%-45% was released under normal physiological conditions. The accelerated drug release in the TME was due to disulfide bond cleavage and ester bond hydrolysis in the presence of GSH and acidic pH, whereas under normal conditions, the NCs remained stable/undissociated. Cellular uptake studies confirmed enhanced BBR uptake in GSH-rich cancer cells (H1975) compared with normal cells (BEAS-2B and HEK293A). Following uptake, compared with the free form of the drug, the optimized nanocarriers displayed significant selective cytotoxicity and apoptosis in cancer cells by notably downregulating anti-oxidant (NFE2L2, HO-1, NQO1, and TXRND1) and anti-apoptotic (MCL-1) genes while upregulating pro-apoptotic genes (PUMA and NOXA). This resulted in increased oxidative stress, thereby inducing selective apoptosis in the GSH-rich lung cancer cells. These results suggest that the synthesized novel NCs hold great potential for specifically delivering drugs to cancer cells (with a reduced environment) while sparing normal cells, thus ensuring safe and efficient cancer therapy.

In Vitro and In-Silico Assessment of Gaussian Curvature-driven Internalization Kinetics of Nanoparticles.

Nanoparticles have been of significant interest in various biomedical domains such as drug delivery, gene delivery, cytotoxicity analysis, and imaging. Despite the synthesis of a variety of nanoparticles, their cellular uptake efficiency remains a substantial obstacle, with only a small fraction of delivered nanoparticles (NPs) have been reported to traverse the cell membrane within 24 h. Consequently, higher doses are often necessitated, leading to increased toxicity concerns. In this investigation, we illustrate that nanoparticles having negative Gaussian curvature demonstrate rapid and efficient internalization into cells by lowering the energy barrier for membrane bending. Specifically, three types of gold nanoparticles; gold nanorods (GNR), gold nanodogbones at pH 4 (GDB4), and gold nanodogbones at pH 6 (GDB6) were synthesized, with Gaussian curvatures of 0, -166.91, and -376.62, respectively. Cellular uptake studies conducted via ICP-OES analysis reveal that GDB6 is taken up 140% more in A549 cells and 77% more in NIH3T3 cells compared to GNR. Confocal microscopy-based uptake studies further confirm the higher uptake of GDB6 compared to GNR. Additionally, molecular simulations indicate that GDB nanoparticles exhibit a significantly larger free energy change during translocation compared to GNR, emphasizing the impact of nanoparticle shape on uptake and translocation through the membrane and validating the efficacy of negative Gaussian curvature in enhancing cellular uptake, consistent with experimental observations. Overall, our findings emphasize the importance of nanoparticle curvature modulation in maximizing cellular uptake efficiency for improved biomedical applications, providing valuable insights into the design of nanomaterials for drug delivery purposes.

Facile Approach to Develop Peptide-Stabilized CdS/CdSe Quantum Dots for Cellular Imaging.

This study presents a mild, one-pot synthetic approach for the synthesis of multicolor, water soluble, photo luminescent CdS and CdSe quantum dots (QDs). To achieve this goal, cyclic peptides containing cysteine residues are rationally designed and synthesized. Among the peptides tested, those containing two cysteine residues exhibit superior stabilizing properties, ensuring the solubility and long-term stability of the QDs in aqueous solutions for several months. The newly synthesized QDs exhibit unique excitation-dependent multicolor photoluminescence with a quantum yield of 20.55% and 45.50% for CdS and CdSe, respectively, providing versatility for imaging applications. Cellular uptake studies using HCT 116 cells reveal effective internalization of the QDs into both the cytoplasm and nucleus, highlighting their potential applications in bioimaging and drug delivery. This green synthesis approach underscores the crucial role of peptide chemistry in nanoparticle stabilization, paving the way for the development of functional nanomaterials tailored for specific uses in bioimaging and nanomedicine.

Tuned Manganese-Impregnated Mesoporous Silica Nanoparticles as a pH-Responsive Dual Imaging Probe.

The limitations of individual imaging modalities have led to significant interest in hybrid imaging methods that combine the advantages of multiple techniques. The development of diverse dual imaging agents, which offer the exceptional sensitivity of single-photon emission computed tomography (SPECT) and the high spatial resolution of magnetic resonance imaging (MRI), has been addressing the demand for more advanced diagnostic pharmaceuticals. In this study, 99mTc-labeled manganese oxide-loaded mesoporous silica nanoparticles (MSNs), conjugated with folic acid as the targeting moiety and the chelating agent H2pentapa-en-NH2 (99mTc-MnOx-MSN-FA-pa), were developed for targeted SPECT-MRI dual imaging. The toxicity of the nanoparticles was confirmed through an MTT assay, showing >90% viability in HEK-293 and MDA-MB-231 cells at concentrations up to 200 μg/mL, indicating nonsignificant toxicity. Cellular uptake studies showed that folic acid functionalization effectively accentuated tumor-specific intracellular uptake of nanoparticles in MDA-MB-231 cells through folate receptor-mediated endocytosis. Additionally, the radiolabeling yield of 99mTc-MnOx-MSN-FA-pa was found to be 99.6 ± 0.8% (n = 3), and the pH-responsive release of paramagnetic manganese ions increased the r1 relaxivity of the nanoprobe to 11.37 mM-1 s-1. In vivo SPECT imaging demonstrated rapid tracer accumulation in MDA-MB-231 xenografts, with a tumor-to-muscle ratio of 6.01 ± 0.51 at 2 h, and minimal uptake in nontargeted organs. In vivo MRI studies indicated the strongest tumor contrast at 2 h postinjection. Given its desirable contrast enhancement in T1 MRI and SPECT imaging, along with low toxicity, MnOx-MSN-FA-pa shows potential as an effective multifunctional nanoprobe for precise tumor imaging.

Bufonis venenum extract loaded novel cholesterol-free liposome for the treatment of hepatocellular carcinoma.

This study aims to improve the solubility and the toxicity of Bufonis venenum, and finally enhance the therapeutic outcomes of hepatocellular carcinoma (HCC). The cholesterol-free liposomes simultaneously encapsulate bufadienolides and indolealkylamines (Non-Cholesterol-Bufonis Venenum Extract-Liposome, Non-Chol-BVE-LP) was prepared by the thin-film evaporation technique. In vitro, the cytotoxicity, cell apoptosis study, cellular uptake and hemolysis studies were evaluated in HepG2 cells. In vivo, the biodistribution and anti-tumor activity studies were conducted in BALB/C mice with HepG2 cells. The liposomes showed good size distribution, encapsulation efficiency drug loading capacity and slower drug release. Non-Chol-BVE-LP had higher cytotoxicity on HepG2 cells and induced more apoptosis on HepG2 Cells compared with BVE. In addition, the liposomes could accumulate in tumor by passive targeting, thus facilitating the anti-tumor effects. In vivo, Non-Chol-BVE-LP showed equivalent anti-tumor efficacy to the first-line anti-HCC drug sorafenib. The study provided new ideas for the development and clinical application of Bufonis venenum related formulation and offered new drug for the treatment of HCC.

Elucidating the uptake and trafficking of nanostructured lipid carriers as delivery systems for miRNA

Cationic nanostructured lipid carriers (cNLCs) represent promising non-viral carriers for nucleic acids, such as miRNAs, forming stable self-assembled miRNA complexes due to electrostatic interactions. Prepared by high-pressure homogenization, cNLC formulations, both with and without Nile Red dye demonstrated stable particle sizes in the range of 100–120 nm and positive surface charges (>30 mV), which are necessary for effective cellular uptake. The miRNA complexes formed at mass ratios of 1:2.5 and 1:5 showed similar stability and size, with positive zeta potentials, as well as high cell viability (> 80 %) in 3T3-L1 and MCF-7 cell lines. The cellular uptake studies of miRNA:cNLC complexes in both cell lines revealed that uptake was time- and concentration-dependent, with rapid initial uptake in 30 min and a zig-zag pattern over 24 h. To elucidate the endocytosis mechanism of miRNA:cNLC complexes, 3T3-L1 and MCF-7 cells were incubated with different inhibitors (chlorpromazine, 5-[N-ethyl-N-isopropyl] amiloride, dynasore, nystatin, or sodium azide with 2-deoxy-d-glucose). Results showed significant inhibition of uptake at low temperatures and with ATP depletion, suggesting endocytosis, particularly macropinocytosis, as the main uptake mechanism in 3T3-L1 cells. In MCF-7 cells, the uptake was less inhibited by the substances, indicating the need for more specific methods to fully decipher the endocytic mechanisms involved. Confocal laser scanning microscopy images revealed that the complexes are internalized in vesicles, and are primarily localized in the juxtanuclear region, suggesting trafficking through the endolysosomal system. Colocalization study with LysoTracker™ Green DND-26 showed significant colocalization of miRNA:cNLC complexes with lysosomes in 3T3-L1 cells, indicating trafficking through the endolysosomal system. In MCF-7 cells, colocalization was lower, suggesting macropinocytosis as the primary uptake mechanism. Additional studies showed partial colocalization between labeled NLCs and miRNA, indicating that about 50 % of miRNA is released from NLCs within 30 min post-transfection.

Bio-Inspired Polymeric Solid Lipid Nanoparticles for siRNA Delivery: Cytotoxicity and Cellular Uptake In Vitro.

Nanomedicine has introduced strategies that provide precise diagnosis and treatment with fewer side effects than traditional therapies. Treatments for neurodegenerative disorders, including Parkinson's disease, are palliative, necessitating an innovative delivery system with a curative function. This study investigated a solid lipid nanoparticle (SLNP) system's ability to bind and safely deliver siRNA in vitro. SLNPS were formulated using sphingomyelin and cholesterol, with Ginkgo biloba leaf extract (GBE) incorporated to enhance biocompatibility and neuroprotection. Poly-L-lysine (PLL) functionalization ensured successful siRNA binding, safe transport, and protection from nuclease degradation. SLNPs were physicochemically characterized, with binding and protection of siRNA assessed using agarose gels. Cytotoxicity, apoptotic induction, and cellular uptake studies were undertaken in the human neuroblastoma (SH-SY5Y) and embryonic kidney (HEK293) cells. The GBE-PLL-SLNPs had an average size of 93.2 nm and demonstrated enhanced binding and protection of the siRNA from enzyme digestion, with minimal cytotoxicity in HEK293 (<10%) and SH-SY5Y cells (<15%). Caspase 3/7 activity was significantly reduced in both cells, while efficient cellular uptake was noted. The present study provided a solid basis as a proof of principle study for future applications of the potential therapeutic in vitro, promising to address the unmet medical needs of patients with neurological disorders.

Sustainable Carbon Dots Loaded into Carboxymethylcellulose Based Hydrogels for Uterine Cancer Bioimaging.

Background/Objectives: The development of innovative materials for disease diagnostics and therapeutics is a fast-growing area of scientific research. In this work, we report the development of innovative hydrogels incorporating carbon dots (Cdots) for bioimaging purposes. Methods: The Cdots were prepared using a sustainable and low-cost process, starting with an underused fiber from the Brazilian semiarid region. Spectroscopy analysis (Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, UV-visible spectroscopy), X-ray diffraction, photoluminescence, zeta potential, scanning electron microscopy, and transmission electron microscopy were used to characterize these hydrogels. In addition, biocompatibility using the resazurin assay and cellular uptake by confocal microscopy were evaluated. Results: Our results showed that the Cdots changed the structure and crystallinity of hydrogels, mainly due to heat treatment. In addition, hydrogels' chemical groups suffer red and blue shifts following the Cdots incorporation. Moreover, the Cdots were homogeneously incorporated into the hydrogel matrix. Importantly, the cytotoxicity levels were maintained above 90% (p < 0.01), and cellular uptake studies using HeLa cells demonstrated intracellular fluorescence of both the Cdots and hydrogels after incubation. Additionally, the concentration of Cdots within hydrogels significantly affected fluorescence intensity, even compared with pure Cdots. Conclusions: These results showcase the potential for these hydrogels to be further developed as biomarkers and therapeutic biomaterials for women's health.

Formulation and Characterization of β-Cyclodextrins-Nitazoxanide Inclusion Complexes: Enhanced Solubility, In Vitro Drug Release, and Antiviral Activity in Vero Cells.

Background/Objectives: Nitazoxanide (NTX) exhibits promising therapeutic potential; its effectiveness is constrained by its low oral bioavailability due to its poor water solubility and limited permeability. Methods: This study focused on developing a complex of NTX with β-cyclodextrins (β-CDs), specifically β-CD and hydroxypropyl-β-cyclodextrin (Hβ-CD), to enhance the solubility and antiviral activity of NTX. Results: The formation of the β-CD:NTX in an aqueous solution was verified using UV-visible spectroscopy, confirming a 1:1 inclusion complex. Characterization of the solid β-CD:NTX complexes was confirmed via FTIR, X-ray diffraction (XRD), scanning electron microscopy (SEM), and DSC-TGA analyses. Molecular docking studies revealed that the NTX thiazole ring with the nitro group was positioned within the β-CDs cavity, while the benzene ring remained outside. Phase solubility tests showed that β-CD:NTX complexes were formed with high stability constants, demonstrating a linear increase in NTX solubility as the β-CD concentration increased. Dissolution tests revealed rapid and nearly complete NTX release within 90 min for β-CD:NTX and Hβ-CD:NTX complexes. The β-CD:NTX complexes were tested for their antiviral activity against Herpes simplex virus (HSV-1) cultures. Results showed that the Hβ-CD:NTX complex had significantly higher antiviral efficacy than β-CD:NTX and free NTX alone. Moreover, cytotoxicity and cellular uptake studies on Vero cells indicated that the Hβ-CD:NTX complex demonstrated lower cytotoxicity and had the highest IC50 value, followed by β-CD:NTX and free NTX. Conclusions: These findings suggest that Hβ-CD:NTX inclusion complexes may serve as effective carriers for delivering NTX in HSV-1 treatments using Vero cell models.

Preparation, Characterization, and Cytotoxicity Study of Nitrogen-Doped Graphene Quantum Dots Functionalized Hyaluronic Acid Loaded with Docetaxel-Catalyzed Nanoparticles for Breast Cancer Imaging and Targeting In Vitro

Many women worldwide are impacted by breast cancer that also claims many lives every year. Docetaxel (DOC) is the natural chemotherapeutic agent used most commonly for the treatment of breast cancer. However, it has some drawbacks including low water solubility, a long half-life, an unregulated rate of discharge from the target site, etc. The objective of our present research reported here was to formulate DOC-loaded hyaluronic acid (HA)-functionalized nitrogen-doped graphene quantum dots (N-GQDs) nanoconjugate (DOC@HA-SDH-NGQDs) by using the adhesive properties of succinic acid dihydrazide (SDH) as a hopeful nanocarrier system for potential breast cancer imaging and targeting. The prepared DOC@HA-SDH-NGQDs nanoparticles (NPs) demonstrated 87.58 % drug loading, 97 % drug release, and −14.5 zeta potential indicating excellent stability. The outcomes of our MTT assay showed the full biocompatibility of the produced nanoconjugate and the significant reduction in MCF7 breast cancer cell viability observed in samples of DOC-loaded HA-SDH-NGQDs NP when we compared to other formulations. The MCF7 cancer cell line was found to respond better to the DOC@HA-SDH-NGQDs nanocomposite than to the standard drug, according to the cellular uptake studies. Conclusively, our described study, we believe, is the first to describe HA-NGQDs nanotherapeutics (NTCs) with the added adhesive qualities of SDH for breast cancer imaging and treatment via pH-triggered targeted anticancer drug delivery.

Ct-DNA compaction by nanoparticles formed by silica and gemini surfactants having hydroxyl group substituted spacers: In vitro, in vivo, and ex vivo gene uptake to cancer cells

Hybrid nanoparticles formed by Silica (SiO2) coated with cationic gemini surfactants with variable hydroxyl group substituted spacers, 12-4(OH)-12,2Br− and 12-4(OH)2-12,2Br− have shown a great extent of compaction of calf thymus DNA (ct-DNA) compared to conventional counterpart cationic surfactant, dodecyl trimethylammonium bromide (DTAB). Study shows not only the hydrophobicity of the spacer but also the hydrogen bonding interactions between the hydroxyl group substituted spacer and DNA have a great role in DNA compaction. 12-4(OH)2-12,2Br− is more efficient in compacting ct-DNA compared to 12-4(OH)-12,2Br− due to the stronger binding of the former with ct-DNA than the latter. While 12-4(OH)-12,2Br− makes 50 % ct-DNA compaction at its 0.63 μM concentration in the presence of SiO2 nanoparticles, the same % of compaction can be achieved at a concentration as low as 0.25 μM of 12-4(OH)2-12,2Br−. However, DTAB makes 50 % ct-DNA compaction at a concentration as high as 7.00 μM under the same condition. Therefore, the present systems address the very common challenge, i.e., cytotoxicity due to cationic surfactants. The system of 12-4(OH)2-12,2Br− coated SiO2 nanoparticles displays the maximum cell viability (≥90 %), causing the least cell death in the mouse fibroblast cells (NIH3T3) cell lines compared to the cell viability of ≤80 % for DTAB. 12-4(OH)2-12,2Br− coated SiO2 nanoparticles system has presented excellent in vitro cellular uptake of genes on mouse mammary gland adenocarcinoma (4T1) cells after incubating for 3 h and 6 h. In vivo study shows that 12-4(OH)2-12,2Br− coated SiO2 nanoparticles system takes the highest amount of ct-DNA in cells and tumors in a time-dependent manner. The ex vivo studies using different organs of the mice demonstrate that the tumor sites in the breast of the mice are most affected by these formulations. Cytotoxicity assays and cellular uptake studies suggest that the present systems can be used for potential applications for gene delivery and oncological therapies.

Novel synthesized seleno-glycoconjugates as cosmeceutical ingredients: Antioxidant activity and in vitro skin permeation

Following recent successful results in the search for innovative semi-synthetic cosmeceutical Se-glycoconjugates, the work reported herein explores the development and evaluation of second-generation selenosugar-linked hydroxycinnamic acids as new potential cosmeceutical ingredients. Utilizing a Pummerer-like rearrangement for C1-acetylation, these novel compounds were synthesized and characterized using NMR spectroscopy and HRMS, confirming their structures and purity. The biological evaluation focused on their radical scavenging preliminary screening, and cytotoxic and antioxidant activities in human immortalized keratinocytes, revealing significant potential for use in skin care formulations aimed at counteracting oxidative stress and promoting skin health. Cellular uptake studies conducted on HaCaT keratinocytes using UHPLC-QqTOF-MS metabolomics demonstrated effective internalization of these compounds, which is crucial for their efficacy as topical agents. Furthermore, percutaneous absorption tests using the Franz diffusion cell method and subsequent HPLC-DAD analysis provided insights into the compound skin permeation capabilities, a critical factor for their practical application in cosmeceuticals.

In vitro cellular uptake and insulin secretion studies on INS-1E cells of exendin-4-loaded self-nanoemulsifying drug delivery systems

Exendin-4 (ex-4) is a peptide molecule that regulates blood glucose levels without causing hypoglycemia by providing insulin secretion from beta cells in the pancreas. Self-nanoemulsifying drug delivery systems (SNEDDS) attract attention for oral administration of therapeutic peptide/proteins because they protect therapeutic peptide/proteins from the gastric environment, reduce changes due to food effects, are easy to prepare and scale-up. Ex-4 has no commercial form that can be administered orally. In this study, the cytotoxicity, cellular uptake, and insulin secretion of ex-4 and ex-4/chymostatin (chym) SNEDDS were investigated on INS-1E rat pancreatic beta cells. The effect of ex-4 and ex-4/chym SNEDDS on cell viability in INS-1E cells increased when the dilution ratio higher. Ex-4 and ex-4/chym SNEDDS increased insulin levels in 2.8 mM (low-dose) glucose-induced INS-1E cells 2.21-fold and 2.17-fold compared to control, respectively. Ex-4 and ex-4/chym SNEDDS increased insulin levels in 16.7 mM (high dose) glucose-induced INS-1E cells compared to control, respectively. In cellular uptake studies, coumarin-6 solution penetrated the apical membrane of INS-1E cells and remained in the cytoplasm, while coumarin-6 loaded SNEDDS were visualized in the nuclei of the cell. These findings will likely be useful in the development of new formulations for the oral administration of peptides/proteins.

Chrysin-loaded Soluplus-TPGS mixed micelles: Optimization, characterization and anticancer activity against hepatocellular carcinoma cell line

Chrysin is a natural flavonoid found in various plants, and it has shown potential therapeutic benefits such as anti-inflammatory, antioxidant, and anticancer properties. However, its clinical application is limited due to poor solubility and low bioavailability. Mixed micelles can help overcome these problems. This study focuses on preparation of Chrysin-loaded mixed micelles with the help of solvent diffusion evaporation method. Soluplus and TPGS were the main components of the formulation, and their proportions were optimized with the help of the central composite design matrix. Thirteen batches were constructed to optimize the mixed micelle formulation based on different Soluplus: Chrysin ratios and TPGS percentages. Response surface methodology and various models were employed to analyze micellar size, size distribution, encapsulation efficiency, and drug loading. The optimized formulation, CHR-MM1, exhibited a particle size of 132.2 ± 7.9 nm, encapsulation efficiency of 98.01 ± 2.27 %, and a zeta potential of −2.10 mV. TEM images revealed that the micelles were spherical in shape. Characterization studies, including FTIR and X-ray diffraction, confirmed the successful encapsulation of Chrysin in an amorphous form within the mixed micelles. The in vitro release studies demonstrated a sustained release behavior in both acidic and neutral pH conditions. The Korsmeyer-Peppas model best described the drug release kinetics. Cellular uptake studies using fluorescence microscopy revealed enhanced internalization of the mixed micelles into Hep G2 cells. Moreover, MTT assays indicated a concentration- and time-dependent cytotoxicity of Chrysin-loaded mixed micelles against Hep G2 cells, outperforming free Chrysin. The flow cytometry analysis results suggested an enhanced apoptotic activity of Chrysin in the mixed micelles as compared to free drug. This study provides a promising strategy for improving the solubility, cellular uptake, and cytotoxicity of Chrysin. Therefore, our results point to the potential of Chrysin-loaded mixed micelles to serve as a therapeutic option for hepatocellular carcinoma.