Chlorin e6 Conjugated Methoxy-Poly(Ethylene Glycol)-Poly(D,L-Lactide) Glutathione Sensitive Micelles for Photodynamic Therapy.

PURPOSE: In this study, we developed a polymeric micellar system for glutathione-mediated intracellular delivery of a photosensitizer, chlorin e6 (Ce6) by synthesizing an amphiphilic polymer, methoxy-poly(ethylene glycol)-poly(D,L-lactide)-disulfide-Ce6 (mPEG-PLA-S-S-Ce6), which self-assembled in aqueous environment to form micelles. METHODS: The polymer-drug conjugate was characterized by NMR. The singlet oxygen (2O1) generation and in vitro release of Ce6 micelles were evaluated. Further, glutathione-mediated intracellular drug delivery was assessed in human alveolar adenocarcinoma cells (A549), mouse mammary carcinoma cells (4 T1) and 3D A549 spheroids. RESULTS: The micellar system protected Ce6 from aggregation leading to improved 2O1 generation compared to free Ce6. Due to the availability of glutathione, the disulfide bonds in the micelles were cleaved resulting in rapid release of Ce6 evident from the in vitro study. The Ce6 micelles displayed quicker drug release in presence of glutathione monoester (GSH-OEt) pre-treated A549 and 4 T1 cells compared to without pre-treated cells. In vitro phototoxicity of micelles displayed enhanced toxicity in 10 mM GSH-OEt pre-treated A549 and 4 T1 cells compared to untreated cells. As anticipated, Ce6 micelles showed lower drug release in presence of 0.1 mM of buthionine sulfoximine (BSO) pretreated A549 and 4 T1 cells exhibiting lower phototoxicity. Further, A549 3D spheroids treated with Ce6 micelles showed significant inhibition in growth, enhanced phototoxicity, and cellular apoptosis in comparison to free Ce6. CONCLUSION: The above results showed that the developed strategy could be effective in improving the PDT efficacy of Ce6, and the developed polymeric micellar system could be utilized as a PDT regimen for cancer.

Transferrin-Modified Vitamin-E/Lipid Based Polymeric Micelles for Improved Tumor Targeting and Anticancer Effect of Curcumin.

PURPOSE: Transferrin receptor (TfR) is up-regulated in various malignant tumors not only to meet the iron requirement, but also to increase the cell survival via participation in various cellular signaling pathways. Here we explored transferrin as ligand for Poly(ethylene Glycol) (PEG)-ylated vitamin-E/lipid (PE) core micelles (VPM). METHODS: Transferrin modified polymer was synthesized and drug loaded micelles were evaluated in 2D Hela and HepG2 cancer cells for cellular uptake and cytotoxicity and in 3D Hela spheroids for growth inhibition, uptake and penetration studies. RESULTS: Targeted (Tf-VPM) and non-targeted (VPM) micelles showed mean hydrodynamic diameter of 114.2 ± 0.64 nm and 117.4 ± 0.72 nm and zeta potential was -22.8 ± 0.62 and -14.8 ± 1.74 mV, respectively. Cellular uptake study indicated that the Tf-CVPM were taken up by cancer cells (Hela and HepG2) with higher efficiency. Enhanced cytotoxicity was demonstrated for Tf-VPM compared to CVPM. Marked spheroid growth inhibition following treatment with Tf-CVPM was observed compared to the treatment with non-targeted CVPM. CONCLUSIONS: The developed transferrin-modified micelles have improved ability to solubilize the loaded drugs and could actively target solid tumors by its interaction with over-expressed transferrin receptors. Therefore, the nano-micelles could be further explored for its potential utilization in cancer therapy.

Isoproterenol-induced cardiac ischemia and fibrosis: Plant-based approaches for intervention.

Heart is the most active and incumbent organ of the body, which maintains blood flow, but due to various pathological reasons, several acute and chronic cardiac complications arise out of which myocardial infarction is one of the teething problems. Isoproterenol (ISP)-induced myocardial ischemia is a classical model to screen the cardioprotective effects of various pharmacological interventions. Phytochemicals present a novel option for treating various human maladies including those of the heart. A large number of plant products and their active ingredients have been screened for efficacy in ameliorating ISP-induced myocardial ischemia including coriander, curcumin, Momordica, quercetin, and Withania somnifera. These phytochemicals constituents may play key role in preventing disease and help in cardiac remodeling. Reactive oxygen species scavenging, antiinflammatory, and modulation of various molecular pathways such as Nrf2, NFкB, p-21 activated kinase 1 (PAK1), and p-smad2/3 signaling modulation have been implicated behind the claimed protection. In this review, we have provided a focused overview on the utility of ISP-induced cardiotoxicity, myocardial ischemia, and cardiac fibrosis for preclinical research. In addition, we have also surveyed molecular mechanism of various plant-based interventions screened for cardioprotective effect in ISP-induced cardiotoxicity, and their probable mechanistic profile is summarized.

Evaluation of Anti-Tumor Efficacy of Vorinostat Encapsulated Self-Assembled Polymeric Micelles in Solid Tumors.

Vorinostat (VOR), a potent HDAC inhibitor, suffers from low solubility and poor absorption, which hinders its successful application in therapy, especially in the treatment of solid tumors. In this study, an effort to improve the physicochemical characteristics of VOR was made by encapsulating it in PEG-PLGA copolymeric micelles. VOR-loaded PEG-PLGA micelles (VOR-PEG-PLGA) were produced by thin-film hydration and physicochemically characterized. The PEG-PLGA micelles had an average size of 124.06 ± 2.6 nm, polydispersity index of 0.27 ± 0.1, and entrapment efficiency of 90 ± 2.1%. Micelles were characterized by TEM, DSC, and drug release studies. The drug release occurred in a sustained manner up to 72 h from PEG-PLGA micelles. In the in vitro cell-based studies using human breast cancer (MDA MB 231) and murine melanoma (B16F10) cell lines, VOR-PEG-PLGA micelles exhibited superior cellular internalization, enhanced cytotoxic activity, and greater apoptosis compared to free drug. Percent cell killing of 54.9% for VOR-PEG-PLGA-treated cells was observed after 24 h compared to 36% for free VOR in MDA MB 231 cell line. Further, significant tumor suppression was witnessed in B16F10 tumor-bearing mice treated with VOR-PEG-PLGA micelles with a 1.78-fold reduction in tumor volume compared to free VOR-treated animals. Overall, the VOR-PEG-PLGA micelles improved the biopharmaceutical properties of VOR, which resulted in enhanced anti-tumor efficacy. Therefore, the newly developed nano-formulation of VOR could be considered as an effective treatment option in solid tumors.

Polylactide-Based Block Copolymeric Micelles Loaded with Chlorin e6 for Photodynamic Therapy: In Vitro Evaluation in Monolayer and 3D Spheroid Models.

Recently, photodynamic therapy (PDT) has found wide application as a noninvasive treatment modality for several cancers. However, the suboptimal delivery of photosensitizers (PSs) to the tumor site is a drawback, which inhibits the effectiveness of PDT. Hydrophobicity, strong oxygen and light dependence, and limited tissue penetrability of photosensitizers represent the major barriers to the clinical application of PDT. In order to improve biopharmaceutical properties of a clinically approved photosensitizer chlorin e6 (Ce6), we developed a nanoformulation encapsulating Ce6 in methoxy-poly(ethylene glycol)-poly(d,l-lactide) (mPEG-PLA) copolymeric micelles. The physicochemical properties, including particle size, zeta potential, encapsulation efficiency, drug loading, generation of reactive oxygen species following near-infrared light illumination (633 nm), and in vitro drug release, were determined. The therapeutic efficacy of Ce6-mPEG-PLA micelles following illumination were evaluated in vitro in both two- and three-dimensional cell culture systems by using human uterine cervix carcinoma (HeLa) and human alveolar adenocarcinoma (A549) cells in monolayers and in A549 spheroids, respectively. The mPEG-PLA micelles were stable with a particle size of 189.6 ± 14.32 nm and loaded Ce6 efficiently (encapsulation efficiency ∼75%). The Ce6-loaded micelles generated singlet oxygen at a higher concentration compared to free Ce6 in aqueous media. Ce6-mPEG-PLA micelle mediated PDT showed improved cellular internalization in both of the cell lines, resulting in enhanced cytotoxicity compared to free Ce6. In contrast, the Ce6-loaded micelles did not show any cytotoxicity in the absence of irradiation. The Ce6-loaded micelles exhibited deep penetration in the spheroids leading to phototoxicity and cellular apoptosis in the A549 spheroidal model. Results from this study indicated that the newly developed nanoformulation of Ce6 could be utilized in PDT as an effective treatment modality for solid tumors.

Curcumin Delivery by Poly(Lactide)-Based Co-Polymeric Micelles: An In Vitro Anticancer Study.

PURPOSE: This work describes the synthesis of block co-polymeric micelles, methoxy-poly(ethylene glycol)-poly(D,L-lactide) (mPEG-PLA) to encapsulate Curcumin (CUR), thereby improving the dispersibility and chemical stability of curcumin, prolonging its cellular uptake and enhancing its bioavailability. METHODS: CUR-mPEG-PLA micelles, was prepared using the thin-film hydration method and evaluated in vitro. The preparation process was optimized with a central composite design (CCD). Micelles were characterized by size, transmission electron microscopy, loading capacity, and critical micelle concentration (CMC). The cytotoxicity of CUR-mPEG-PLA micelles was investigated against murine melanoma cells, B16F10 and human breast cancer cells, MDA-MB-231. RESULTS: The average size of the CUR-mPEG-PLA micelles was 110 ± 5 nm with polydispersity index in the range of 0.15-0.31, and the encapsulating efficiency for CUR was 91.89 ± 1.2, and 11.06 ± 0.8% for drug-loading. Sustained release of CUR from micelles was observed with 9.73% CUR release from micelles compared to 64.24% release of free curcumin in first 6 h under sink condition. The CUR-mPEG-PLA was efficiently taken up by the cancer cells, B16F10 and MDA-MB-231. Following 24 h incubation, CUR-mPEG-PLA induced higher cytotoxicity compared to free CUR in MDA-MB-231 cell lines indicating exposure of higher dose of free CUR to cells lead to up-regulation of drug efflux mechanisms leading to decreased cell death in case of free CUR administration. CONCLUSION: Our results indicate that the proposed micellar system has the potential to serve as an efficient carrier for CUR by effectively solubilizing, stabilizing and delivering the drug in a controlled manner to the cancer cells.

Revolutionizing Dentistry: The Applications of Artificial Intelligence in Dental Health Care.

Artificial intelligence (AI) is transforming the landscape of health care, and dentistry is no exception. This article explores the various applications of AI in dentistry, showcasing how this advanced technology is revolutionizing diagnosis, treatment, and patient care. From enabling early detection of oral diseases to enhancing the precision of dental procedures, AI is driving the industry toward more efficient and effective dental healthcare services. This article delves into the specific ways in which AI is being integrated into dental practices, highlighting its potential to improve patient outcomes and advance the field of dentistry.

Lipid and poly (ethylene glycol)-conjugated bi-functionalized chlorine e6 micelles for NIR-light induced photodynamic therapy.

BACKGROUND: To develop a photosensitizer, chlorin e6 (Ce6)-based amphiphilic polymer, DP-Ce6, where DOPE and PEG are conjugated to Ce6, which would self-assemble to form polymeric micelles (DP-Ce6-M) in aqueous environment. METHODS: DP-Ce6-M were characterized for particle size, zeta potential, and singlet oxygen (1O2) generation. Cellular internalization, phototoxicity were investigated against monolayer and 3D spheroids of human lung adenocarcinoma cells (A549). RESULTS AND CONCLUSIONS: DP-Ce6-M formed stable micelles with particles size of 58.2 ±â€¯1.6 nm. Solubility of Ce6 was improved. Photoactivity of DP-Ce6-M was sustained in regard to 1O2 generation compared to free Ce6. The DP-Ce6-M showed enhanced internalization and growth inhibition in monolayer and spheroidal cells. Overall, DP-Ce6-M demonstrated the potential for further exploration as PDT agent for cancer treatment.

Biotin functionalized PEGylated poly(amidoamine) dendrimer conjugate for active targeting of paclitaxel in cancer.

In the current study, we employed poly(amidoamine) (PAMAM) dendrimers of generation 4 (G4) to deliver paclitaxel (PTX), a poorly soluble anti-cancer agent precisely to cancer cells via its conjugation on dendrimer surface. Further, G4 PAMAM has been PEGylated (PEG) and tagged with Biotin, an essential micronutrient for cellular functions, receptors of which are overexpressed in certain cancers. The synthesized multifunctional conjugates were characterized by 1H NMR and zeta potential analysis techniques. In addition, the conjugates were evaluated in vitro in cell monolayers and 3D spheroids of biotin receptor over-expressed A549 cell line (human non-small cell lung cancer). G4 PTX PEG-Biotin conjugate penetrated at significantly higher extent in monolayers as well as spheroids as studied by flow cytometry and confocal microscopy by visualizing the cells at varied depth. The G4 PTX PEG-Biotin conjugate demonstrated higher cytotoxicity compared to free PTX and G4 PTX PEG conjugate as assessed by MTT assay in monolayers and Presto Blue assay in detached spheroidal cells. G4 PTX PEG-Biotin demonstrated significant inhibition of growth of tumor spheroids. Therefore, the newly synthesized biotin anchored PTX-conjugated dendrimer system is promising and could be further explored for efficiently delivering PTX to biotin receptor overexpressed cancers.

Development of chlorin e6-conjugated poly(ethylene glycol)-poly(d,l-lactide) nanoparticles for photodynamic therapy.

AIM: In this study, we developed a chlorin e6-conjugated methoxy-poly(ethylene glycol)-poly(d,l-lactide) (mPEG-PLA-Ce6) amphiphilic polymer, which self-assembled to form stable nanoparticles. MATERIALS & METHODS: The nanoparticles were characterized for particle size, ζ-potential and singlet oxygen (1O2) generation. Cellular internalization and phototoxicity were investigated against monolayer and 3D spheroids of human lung adenocarcinoma cells (A549). RESULTS & CONCLUSION: mPEG-PLA-Ce6 exhibited a size of 149.72 ± 3.51 nm and ζ-potential of -24.82 ± 2.94 mV. The 1O2 generation by mPEG-PLA-Ce6 in water was considerably higher than free chlorin e6. The nanoparticles showed enhanced cellular internalization and phototoxicity in monolayer and 3D spheroids. The developed mPEG-PLA-Ce6 has potential application as a nanocarrier of chlorin e6 for photodynamic therapy of solid tumors.

Transferrin-anchored poly(lactide) based micelles to improve anticancer activity of curcumin in hepatic and cervical cancer cell monolayers and 3D spheroids.

In recent years, actively targeted drug delivery systems have been utilized in pre-clinical studies for site-specific delivery of drugs, which reduces toxicities associated with chemotherapy. This study reports the preparation of the tumor homing ligand, transferrin (Tf) anchored methoxy-polyethylene glycol-poly(d,l-Lactide) polymeric micelles (Tf-PP). Curcumin which possess wide anti-cancer activity was loaded into the micelles. Tf-PPC with average particle size of 132.16 ±â€¯1.37 nm and encapsulation efficiency of 88.27 ±â€¯2.53% showed a sustained drug release. The efficacy of Tf-PPC was studied in vitro in Tf-overexpressing human cervical carcinoma (HeLa) and human hepatoma (HepG2) cells. The mouse embryo fibroblast (NIH-3T3) cells were used as control cells. Tf-PPC showed higher internalization compared to non-targeted micelles (PPC). The curcumin-mediated cytotoxicity increased significantly following Tf-PPC treatment in both the tested cell lines. In NIH-3T3 cells, Tf conjugation did not differ in comparison to the non-targeted micelles. Further, the efficiency of Tf-PPC was studied in three-dimensional (3D) HeLa tumor spheroids. The Tf-PPC was efficiently internalized by the spheroidal structures, causing higher cytotoxicity and apoptosis compared to PPC. These results reveal that the newly developed, Tf-PPC could be employed as an effective chemotherapy in the treatment of Tf- overexpressing cancers.

Cholesterol and vitamin E-conjugated PEGylated polymeric micelles for efficient delivery and enhanced anticancer activity of curcumin: evaluation in 2D monolayers and 3D spheroids.

A newly synthesized PEGylated cholesterol/α-tocopheryl succinate (α-TOS) linked polymer (CV) was self-assembled and loaded with curcumin to form a micellar system (C-CVM). The tri-functionalized amphiphilic polymer was constituted of hydrophobic cholesterol and α-TOS connected to hydrophilic PEG via a lysine linker. The synthesized polymer and the micelles were characterized by 1H NMR, DLS, zeta potentiometer, TEM, CMC determination and hemolysis studies. CVM displayed low CMC value of 15 µM with extent of hemolysis as less than 4%. The stable C-CVM with optimum % drug loading (14.2 ± 0.24) displayed Z average of 175.8 ± 0.68 nm with PDI (0.248 ± 0.075) and released curcumin in sustained manner in the in vitro drug release study. C-CVM demonstrated dose-dependent cellular uptake and cytotoxicity in murine melanoma, B16F10 and human breast cancer, MDA-MB-231 cell lines. CV exhibited marked reversal of drug resistance as indicated by significantly higher retention of P-glycoprotein substrate, rhodamine-123 in the resistant B16F10 cell line compared to standard P-glycoprotein inhibitor, verapamil. C-CVM demonstrated significantly higher spheroidal growth inhibition compared to C-PPM. The results provide strong evidence for CVM as promising drug delivery system and confirm the potential of C-CVM as chemotherapy in cancer.

d-α-Tocopheryl Succinate/Phosphatidyl Ethanolamine Conjugated Amphiphilic Polymer-Based Nanomicellar System for the Efficient Delivery of Curcumin and To Overcome Multiple Drug Resistance in Cancer.

Nanomedicines have emerged as a promising treatment strategy for cancer. Multiple drug resistance due to overexpression of various drug efflux transporters and upregulation of apoptotic inhibitory pathways in cancer cells are major barriers that limit the success of chemotherapy. Here, we developed a d-α-tocopherol (α-TOS)/lipid-based copolymeric nanomicellar system (VPM) by conjugating phosphatidyl ethanolamine (PE) and α-TOS with poly(ethylene glycol) (PEG) via an amino acid linkage. The synthesized polymers were characterized by Fourier transform IR, gas-phase chromatography, and 1H and 13C NMR spectroscopy. VPM exhibited mean hydrodynamic diameter of 141.0 ± 0.94 nm with low critical micelles concentrations (CMC) of 15 µM compared to plain PEG-PE micelles (PPM) with size of 23.9 ± 0.34 nm and CMC 20 µM. The bigger hydrophobic compartment in VPM resulted in improved loading of a potent chemotherapeutic drug, curcumin (Cur), and increased encapsulation efficiency (EE) (% drug loading 98.3 ± 1.92, and 85.3 ± 3.29; EE 14.8 ± 0.16 and 12.8 ± 0.09 for VPM and PPM, respectively). Curcumin loaded Vitamin E based micelles exhibited higher cytotoxicity compared to Curcumin loaded PEG-PE micelles in tested cancer cell lines. C-VPM demonstrated ∼3.2 and ∼2.7-fold higher ability to reverse multiple drug resistance compared to PPM and verapamil (concentration used 30 µM), respectively. In the in vivo study by using B16F10 implanted C57Bl6/J mice, C-VPM reduced the tumor volume and weight more efficiently than C-PPM by inducing apoptosis as analyzed by TUNEL assay on tumor cryosections. The newly developed polymeric micelles, VPM with improved drug loadability and ability to reverse the drug resistance could successfully be utilized as a nanocarrier system for hydrophobic chemotherapeutic agents for the treatment of drug-resistant solid tumors.

Polymeric micelles of suberoylanilide hydroxamic acid to enhance the anticancer potential in vitro and in vivo.

AIM: To improve the bioavailability and anticancer potential of suberoylanilide hydroxamic acid (SAHA) by developing a drug-loaded polymeric nanomicellar system. METHODS: SAHA-loaded Poly(ethylene glycol)-block-poly(caprolactone) (PEG-PCL) micelles were developed, and physico-chemically characterized. In vitro cellular uptake, viability and apoptosis-inducing ability of the SAHA-PEG-PCL micelles were investigated. In vivo anticancer activity was evaluated in C57BL/6 mice-bearing tumor. RESULTS: The SAHA-PEG-PCL micelles had optimum size (∼130 nm) with an entrapment efficiency of approximately 67%. The SAHA-PEG-PCL induced stronger cell cycle arrest in G2/M phase leading to higher rate of apoptosis compared to free SAHA. SAHA-PEG-PCL demonstrated significant tumor suppression compared to free SAHA in vivo. CONCLUSION: The physicochemical properties and the antitumor efficacy of SAHA were improved by encapsulating in polymeric micelles.

Cholesterol-conjugated poly(D, L-lactide)-based micelles as a nanocarrier system for effective delivery of curcumin in cancer therapy.

Polymeric micelles have been widely explored preclinically as suitable delivery systems for poorly soluble chemotherapeutic drugs in cancer therapy. The present study reported the development of cholesterol (Ch)-conjugated poly(D,L-Lactide) (PLA)-based polymeric micelles (mPEG-PLA-Ch) for effective encapsulation and delivery of curcumin (CUR) at the tumor site. Cholesterol conjugation dramatically affected the particle size and improved drug loading (DL) and encapsulation efficiency (EE). mPEG-PLA-Ch-CUR showed bigger hydrodynamic diameter (104.6 ± 2.1 nm, and 169.3 ± 1.52 nm for mPEG-PLA and mPEG-PLA-Ch, respectively) due to increased size of the hydrophobic core. The newly developed polymer exhibited low critical micelles concentration (CMC) (25 µg/mL) which is close to lipid-based polymer, PEG-phosphatidyl ethanolamine (12.5 µg/mL) compared to mPEG-PLA (50 µg/mL). mPEG-PLA-Ch micelles exhibited relatively higher EE (93.74 ± 1.6%) and DL (11.86 ± 0.8%) compared to mPEG-PLA micelles (EE 91.89 ± 1.2% and DL 11.06 ± 0.8%). mPEG-PLA-Ch micelles were internalized by the cancer cells effectively and exhibited higher cytotoxicity compared to free CUR in both, murine melanoma (B16F10) and human breast cancer (MDA-MB-231) cells. mPEG-PLA-Ch exhibited satisfactory hemocompatibility indicating their potential for systemic application. Further, mPEG-PLA-Ch-CUR demonstrated higher rate of reduction of tumor volume in B16F10-xenografted tumor-bearing mice compared to free CUR. At the end of 22 days, the tumor reduced to 1.87-fold (627.72 ± 0.9 mm3 versus 1174.68 ± 1.64 mm3) compared to the treatment with free CUR. In conclusion, the experimental data in vitro and in vivo indicated that the newly developed CUR-mPEG-PLA-Ch micelles may have promising applications in solid tumors.

Recent advances in polymeric micelles for anti-cancer drug delivery.

Block co-polymeric micelles receive increased attention due to their ability to load therapeutics, deliver the cargo to the site of action, improve the pharmacokinetic of the loaded drug and reduce off-target cytotoxicity. While polymeric micelles can be developed with improved drug loading capabilities by modulating hydrophobicity and hydrophilicity of the micelle forming block co-polymers, they can also be successfully cancer targeted by surface modifying with tumor-homing ligands. However, maintenance of the integrity of the self-assembled system in the circulation and disassembly for drug release at the site of drug action remain a challenge. Therefore, stimuli-responsive polymeric micelles for on demand drug delivery with minimal off-target effect has been developed and extensively investigated to assess their sensitivity. This review focuses on discussing various polymeric micelles currently utilized for the delivery of chemotherapeutic drugs. Designs of various stimuli-sensitive micelles that are able to control drug release in response to specific stimuli, either endogenous or exogenous have been delineated.