An efficient method for cell sheet bioengineering from rBMSCs on thermo-responsive PCL-PEG-PCL copolymer.

Utilizing both medium enrichment and a thermos-responsive substrate to maintain the cell-to-cell junctions and extracellular matrix (ECM) intact, cell sheet technology has emerged as a ground-breaking approach. Investigating the possibility of using sodium selenite (as medium supplementation) and PCL-PEG-PCL (as vessel coating substrate) in the formation of the sheets from rat bone marrow-derived mesenchymal stem cells (rBMSCs) was the main goal of the present study. To this end, first, Polycaprolactone-co-Poly (ethylene glycol)-co-Polycaprolactone triblock copolymer (PCEC) was prepared by ring-opening copolymerization method and characterized by FTIR, 1 H NMR, and GPC. The sol-gel-sol phase transition temperature of the PCEC aqueous solutions with various concentrations was either measured. Next, rBMSCs were cultured on the PCEC, and let be expanded in five different media containing vitamin C (50 µg/ml), sodium selenite (0.1 µM), vitamin C and sodium selenite (50 µg/ml + 0.1 µM), Trolox, and routine medium. The proliferation of the cells exposed to each material was evaluated. Produced cell sheets were harvested from the polymer surface by temperature reduction and phenotypically analyzed via an inverted microscope, hematoxylin and eosin (H&E) staining, and field emission scanning electron microscopy (FESEM). Through the molecular level, the expression of the stemness-related genes (Sox2, Oct-4, Nanog), selenium-dependent enzymes (TRX, GPX-1), and aging regulator gene (Sirt1) were measured by q RT-PCR. Senescence in cell sheets was checked by beta-galactosidase assay. The results declared the improved ability of the rBMSCs for osteogenesis and adipogenesis in the presence of antioxidants vitamin C, sodium selenite, and Trolox in growth media. The data indicated that in the presence of vitamin C and sodium selenite, the quality of the cell sheet was risen by reducing the number of senescent cells and high transcription of the stemness genes. Monolayers produced by sodium selenite was in higher-quality than the ones produced by vitamin C.

Synthesis, characterization, and evaluation of curcumin-loaded endodontic reparative material.

Curcumin (CUR) is an ancient therapeutic agent with remarkable antimicrobial and anti-inflammatory properties. The purpose of the current study was to synthesize and evaluate a curcumin-based reparative endodontic material to reduce infection and inflammation besides the induction of mineralization during the healing of the dentin-pulp complex. Poly-ɛ-caprolactone (PCL)/gelatin (Gel)/CUR scaffold was synthesized and assessed by scanning electron microscopy, Fourier transform infrared spectroscopy, and thermo-gravimetric analysis (TGA). Agar diffusion test was performed against E. coli, A. baumannii, P. aeruginosa, S. aureus, E. faecalis, and S. mutans. Moreover, proliferative, antioxidative, anti-inflammatory, and calcification properties of these scaffolds on human dental pulp stem cells (hDPSCs) were evaluated. The results showed that PCL/Gel/CUR scaffold had antibacterial effects. Also, these CUR-based scaffolds had significant inhibitory effects on the expression of tumor necrosis factor α and DCF from inflamed hDPSCs (p < 0.05). Moreover, the induction of mineralization in hDPSCs significantly increased after seeding on CUR-based scaffolds (p < 0.05). Based on these findings, the investigated CUR-loaded material was fabricated successfully and provided an appropriate structure for the attachment and proliferation of hDPSCs. It was found that these scaffolds had antimicrobial, antioxidant, and anti-inflammatory characteristics and could induce mineralization in hDPSCs, which is essential for healing and repairing the injured dentin-pulp complex.

Facile fabricating of rGO and Au/rGO nanocomposites using Brassica oleracea var. gongylodes biomass for non-invasive approach in cancer therapy.

In this study, we report a facile green-synthesis route for the fabrication of reduced graphene oxide (rGO) using biomass of Brassica oleracea var. gongylodes (B. oleracea). In addition, we have attempted to provide a green synthesis approach to prepare Gold nanoparticles (Au NPs) on the surface of rGO by using stem extract of B. oleracea. The synthesized Au/rGO nanocomposite was evaluated using UV-visible and FTIR spectroscopy, XRD, Raman, FE-SEM, EDX, AFM and DLS techniques. The obtained results demonstrated that the synthesized Au NPs on the surface of rGO was spherical with sizes ranging about 12-18 nm. The Au/rGO NC was, also, developed as photo-synthesizer system for the photothermal therapy (PTT) of MCF7 breast cancer cells. The near-infrared (NIR) photothermal properties of Au/rGO NCs was evaluated using a continuous laser at 808 nm with power densities of 1 W.cm-2. Their photothermal efficacy on MCF7 breast cancer cells after optimizing the proper concentration of the NCs were evaluated by MTT assay, Cell cycle and DAPI staining. In addition, the potential of the synthesized Au/rGO NCs on reactive oxygen species generating and antioxidant activity were assessed by DPPH. Au/rGO NCs possess high capacity to light-to-heat conversion for absorption in range NIR light, and it is able to therapeutic effects on MCF7 cells at a low concentration. The maximum amount of cell death is 40.12% which was observed in treatment groups that received a combination of Au/rGO NCs and laser irradiation. The results demonstrate that the nanomaterials synthesized by green approach lead to efficient destruction of cancer cell and might thus serve as an excellent theranostic agent in Photothermal therapy applications.

Green and facile synthesis of gold/perlite nanocomposite using Allium Fistulosum L. for photothermal application.

Photothermal therapy (PTT) procedure is anticipated as a new generation of cancer therapy techniques. With this in mind, in this work, an effective drug-free approach was developed to kill MCF7 breast cancer cells using PTT. A novel biocompatible nanocomposite as a PTT transducer was prepared from the in situ phytosynthesis of gold nanoparticles (Au NPs) in the presence of perlite as a platform and extract of Allium Fistulosum L. as a stabilizing and reducing agent (Au/perlite NC). The common characterization techniques such as Fourier transform infrared (FT-IR), zeta potential, dynamic light scattering (DLS), X-ray diffraction (XRD), ultraviolet-visible (UV-vis), scanning electron microscopy (SEM), and energy-dispersive X-ray (EDX) were used to approve the synthesis of Au/perlite NC. The potential of the synthesized NCs on ROS generating and antioxidant activity was assessed by DPPH. In the following, the PTT efficacy of the Au/perlite NC on the destruction of MCF-7 breast cancer cells was assessed in vitro via the cell cycle, cell viability, and DAPI staining assays. The DPPH assay results showed that Au/perlite NC had a radicals scavenging capacity of about 41.47 % in 30 min. Cellular uptake results indicated a significant cell uptake after 1.5 h exposure with Au/perlite NC. Interestingly, cell death was increased dramatically by increasing irradiation time from 6 to 10 min. Cell viability assay revealed that the maximum number of cell death is around 50 % which was observed in the presence of Au/perlite NC by irradiation time of 10 min. Cell cycle results showed that the maximum amount of apoptotic cells (85 %) was observed in Au/perlite NC treatment group received laser irradiation for 10 min. The outcomes demonstrated that the Au/perlite NC can be used as a new drug-free and efficient agent for PTT of breast cancer cells without any concern cytotoxicity.

A Lysine-Functionalized Graphene Oxide-Based Nanoplatform for Delivery of Fluorouracil to A549 Human Lung Cancer Cells: A Comparative Study.

Background: Today, increasing attention is being paid to the application of biocompatible polymers as drug carriers with low cytotoxicity in drug delivery systems to enhance the therapeutic effects of anticancer agents. Materials and Methods: In this study, a biocompatible synthetic polymer (grafted on graphene oxide), composed of N-isopropylacrylamide and 1-vinyl-2-pyrrolidone with L-lysine segments (Lys/PNIPAM-PVP/GO), was developed as a nano-vehicle for the drug. This platform was used for the delivery of fluorouracil (FU) to A549 human lung cancer cells. The superior characteristics of the platform included low-cost precursors, easy synthesis, and the presence of many functional groups for loading drugs. To determine and compare the cytotoxic effects of free FU and its formulated form on the A549 cells, MTT assay was performed; the results showed no significant toxicity difference between the two treated groups (free and formulated FU). For further evaluations, cellular uptake assays were performed via fluorescence microscopy and flow cytometry. Results: Both analyses revealed the low internalization of nano-vehicle into the A549 cells, with 4.31% and 8.75% cellular uptakes in the first two and four hours of treatment. Therefore, the low penetration rate reduced the toxicity of drug-loaded nano-vehicle. Conclusion: Finally, DAPI staining and Annexin V-FITC staining were performed as complementary techniques to determine cell apoptosis.

Comparison study on the effect of gold nanoparticles shape in the forms of star, hallow, cage, rods, and Si-Au and Fe-Au core-shell on photothermal cancer treatment.

Gold nanoparticles (GNPs) indicate potential in the development of cancer treatments as vehicles for thermal damage of cancer cells because of their photothermal heating capability. Herein, we aim to investigate the effect of GNPs geometry as photothermal transducers on cellular uptake and photothermal therapy (PTT) efficacy. For this aim, seven different shapes of anisotropic GNPs: stars, hollow, rods, cages, spheres, Fe-Au, and Si-Au core shells were synthesized and investigate the effect of shape on GNPs optical properties. The physic-chemical characterization of prepared GNPs was investigated by UV-vis, DLS-Zeta, and TEM analysis. The effect of GNPs geometry on cellular uptake was investigated by ICP-MS and flow cytometry method. The PTT potential of these GNPs was compared on MCF7 cells in vitro using MTT assay, cell cycle, and Annexin-V apoptosis assay. While all these GNPs could absorb and convert near-infrared light into heat, gold nanostars exhibited the lowest cytotoxicity, highest cellular uptake and highest heat generation compared to other structures. Following photothermal treatment, due to substantial heat production in MCF7 cells, the apoptosis induction rate was greatly increased for all anisotropic gold nanostructures (stars, hollow, rods, and cages) especially gold nanostars. Combined, we can conclude that GNPs geometry affects cellular uptake and heat generation amount as well as cell destruction by apoptosis pathway. The gold nanostar is promising candidates for photothermal destruction.

Novel Chemo-Photothermal Therapy in Breast Cancer Using Methotrexate-Loaded Folic Acid Conjugated Au@SiO2 Nanoparticles.

Low level laser therapy (LLLT) is known as a safe type of phototherapy to target tumor tissue/cells. Besides, using targeted nanoparticles increases the successfulness of cancer therapy. This study was designed for investigating the combined effect of folate (FA)/Methotrexate (MTX) loaded silica coated gold (Au@SiO2) nanoparticles (NPs) and LLLT on the fight against breast cancer.NPs were synthesized and characterized using FTIR, TEM and DLS-Zeta. The NPs had spherical morphology with mean diameter of around 25 nm and positive charge (+13.3 mV) while after conjugation with FA and MTX their net charge reduced to around -19.7 mV.Our findings in cell uptake studies clearly showed enhanced cellular uptake of NPs after FA and MTX loaded NPs in both breast cancer cell lines especially on MDA-MB-231 due to high expression of folate receptors. The results indicated that LLLT had a proliferative effect on both breast cancer cell lines but in the presence of engineered breast cancer targeted nanoparticle, the efficacy of combination chemo-photothermal therapy was significantly increased using MTT assay (p<0.05), DAPI staining, and cell cycle findings. The highest apoptotic effect on breast cancer cell lines was observed in the cells exposed to a combination of MTX-FA loaded Au@SiO2 NP and LLLT proved by DAPI staining and cell cycle(by increasing the cell arrest in subG0/G1). Taken together a combination of chemotherapy and LLLT improves the potential of breast cancer therapy with minimum side effects.

PLA-PCL-PEG-PCL-PLA based micelles for improving the ocular permeability of dexamethasone: development, characterization, and in vitro evaluation.

The aim of the present research was to investigate the feasibility of developing polylactide-polycaprolactone-polyethylene glycol-polycaprolactone-polylactide (PLA-PCL-PEG-PCL-PLA) based micelles to improve ocular permeability of dexamethasone (DEX). PLA-PCL-PEG-PCL-PLA copolymers were synthesized by a ring-opening polymerization method. DEX was loaded into the developed copolymers. The DEX-loaded micelles were characterized using transmission electron microscopy (TEM) and dynamic light scattering (DLS) methods. Cytotoxicity of the micelles obtained was investigated on L929 cell line. Cellular uptake was followed by fluorescence microscopy and flow cytometry analyses. The release behavior of DEX from the micelles as well as the drug release kinetics was studied. Corneal permeability was also evaluated using an ex vivo bovine model. The pentablock copolymers were successfully synthesized. The TEM results verified the formation of spherical micelles, the sizes of which was approximately 65 nm. The micelles exhibited suitable compatibility on L929 cells. The release profile showed an initial burst release phase followed by a sustained release phase, the kinetic of which was close to the Weibull's distribution model. The micelles showed higher corneal permeability in comparison to a marketed DEX eye drop. Taken together, the results indicated that the PLA-PCL-PEG-PCL-PLA micelles could be appropriate candidates for the ocular delivery of DEX, and probably other hydrophobic drugs.

A simple strategy for chemo-photothermal ablation of breast cancer cells by novel smart gold nanoparticles.

Combined therapeutics dependent on the synergistic effect between photothermal therapy (PTT) and chemotherapy have been anticipated to be the next generation of cancer treatment. For this purpose, a novel gold hydrosol was synthesized by a one-pot approach using poly (2-(dimethylamino) ethyl methacrylate-co- N-isopropylacrylamide-co- 2-vinylpyridine) P(DMAEMA-co-NIPAAM-co-VP) terpolymers as a reducing and stabilizing agent. The synthesized gold hydrosol was recoated by a novel carboxylic acid rich poly (N-isopropyl acrylamide-co-maleic anhydride-graft-citric acid) (PNIPAAm-co-PMA-g-CA) copolymer. Then the gold nanoparticles conjugated with Doxorubicin (DOX) and served as a smart photo synthesizer for chemo/photothermal therapy of MCF-7 breast cancer cells. The synthesized nanoparticles had mono-dispersed spherical morphology with a diameter below 30 nm. Our results from cellular uptake indicated that around 100% of the particles were uptake by MCF-7 cells in the first 3 h of exposure time. The temperature of nanocomposites considerably went up to 45 °C with 10 min exposure to near-infrared irradiation. As a striking result, a single round of PTT combined with a sub-therapeutic dose of DOX revealed a synergistic effect with an ability of robust anti-tumor response up to 99.99% (sum of the population of apoptotic and necrotic cells) demonstrated by Annexin-V, cell cycle and DAPI staining techniques. In conclusion, Near-infrared-mediated photothermal conversion exhibited high effectiveness of a combinational chemo-thermotherapy to treat cancer cells.

A novel gold-based stimuli-responsive theranostic nanomedicine for chemo-photothermal therapy of solid tumors.

The chemo-photothermal therapy performance of a novel theranostic nanoparticles that fabricated through the conjugation of HS-poly(ε-caprolactone)-block-poly(N-isopropylacrylamide)-block-poly(acrylic acid) (HS-PCL-b-PNIPAAm-b-PAA) and gold nanoparticles (GNPs) was extensively investigated. The GNPs@polymer conjugate theranostic NPs was loaded with doxorubicin hydrochloride (DOX) as an anticancer drug through electrostatic interactions to afford GNPs@polymer-DOX theranostic nanomedicine. Temperature and pH-triggered in vitro drug release behavior of the developed theranostic nanomedicine were also examined. The biocompatibility of the synthesized GNPs@polymer theranostic NPs was confirmed through the assessing survival rate of breast cancer cell line (MCF7) using MTT assay. In vitro cytotoxic effects of the GNPs@polymer-DOX theranostic nanomedicine was also evaluated against MCF7 cells in both with or without laser irradiation (532 nm, 145 mJ per pulse, 5 min) conditions, and the results obtained were compared with free DOX as the reference. As the results, the developed GNPs@polymer-DOX can be considered as theranostic nanomedicine for chemo-photothermal therapy of solid tumors.

A novel non-invasive strategy for low-level laser-induced cancer therapy by using new Ag/ZnO and Nd/ZnO functionalized reduced graphene oxide nanocomposites.

In the present research, an effective drug-free approach was developed to kill MCF7 breast cancer cells using low-level laser therapy (LLLT) combined with reduced graphene oxide (rGO)-based hybrid nanocomposites (NCs). Here, fruit extract of Rosa canina was used for the first time to obtain the rGO/ZnO, Ag-ZnO/rGO and Nd-ZnO/rGO NCs by green synthesis. Physico/photochemical properties of these NCs were evaluated using FTIR, XRD, Raman, XPS, SEM/EDX, UV-Vis, DLS and AFM. The potential of the as-synthesized NCs on ROS generating and antioxidant activity were assessed by DPPH. After optimizing the proper concentration of the NCs their anti-tumoral efficacy were evaluated by DAPI staining and MTT assay tests for laser therapy on MCF7 breast cancer cells. Interestingly, cell death was increased dramatically by increasing irradiation dose from 8-32 J/cm2 and then decreased by enhancing laser dose. The maximum amount of cell death is 50% which was observed in the presence of ZnO/rGO 20% by irradiation dose of 32 J/cm2. Furthermore, in comparison with 810 nm, 630 nm lasers were more effective in LLLT of MCF7 cells. The results show the potential of using rGO-based NCs in LLLT, which may be combined with other therapeutic approaches to assist our fight against cancer.

Metformin enhances doxorubicin sensitivity via inhibition of doxorubicin efflux in P-gp-overexpressing MCF-7 cells.

Resistance against chemotherapy is still a major problem in successful cancer treatment in the clinic. Therefore, identifying new compounds with lower side-effects and higher efficacy is an important approach to overcome multidrug resistance (MDR). Here, we investigated the activity and possible mechanism of the antidiabetic drug, metformin, in human doxorubicin (DOX)-resistant breast cancer (MCF-7/DOX) cells. The effect of metformin on the cytotoxicity of DOX was evaluated by MTT assay. The P-gp mRNA/protein expression levels following treatment with metformin were determined using real-time polymerase chain reaction and Western blot analysis, respectively. Intracellular rhodamine 123 accumulation assay was performed to evaluate the P-gp function. Cellular ATP content was determined using ATP assay kit. The effect of metformin on DOX-induced apoptosis was evaluated by annexin V/FITC assay. Exposure to metformin considerably enhanced the cytotoxicity of DOX. Metformin had no substantial effect on P-gp expression, while the activity of P-gp and intracellular ATP content decreased with metformin treatment in a dose-dependent manner. Furthermore, metformin significantly increased the DOX-induced apoptosis. These results indicate that metformin could reverse MDR in breast cancer cells by reducing P-gp activity. Therefore, metformin can be suggested as a potent adjuvant in breast cancer chemotherapy.

Recent progress in theranostic applications of hybrid gold nanoparticles.

A significant area of research is theranostic applications of nanoparticles, which involves efforts to improve delivery and reduce side effects. Accordingly, the introduction of a safe, effective, and, most importantly, renewable strategy to target, deliver and image disease cells is important. This state-of-the-art review focuses on studies done from 2013 to 2016 regarding the development of hybrid gold nanoparticles as theranostic agents in the diagnosis and treatment of cancer and infectious disease. Several syntheses (chemical and green) methods of gold nanoparticles and their applications in imaging, targeting, and delivery are reviewed; their photothermal efficiency is discussed as is the toxicity of gold nanoparticles. Owing to the unique characterizations of hybrid gold nanoparticles and their potential to be developed as multifunctional, we predict they will present an undeniable role in clinical studies and provide treatment platforms for various diseases. Thus, their clearance and interactions with extra- and intra-cellular molecules need to be considered in future projects.