Co-Delivery of Curcumin and Chrysin by Polymeric Nanoparticles Inhibit Synergistically Growth and hTERT Gene Expression in Human Colorectal Cancer Cells.

Nanoparticle (NP)-based combinational chemotherapy has been proposed as a potent approach for improving intracellular drug concentrations and attaining synergistic effects in colorectal cancer therapy. Here, two well-known herbal substances, Curcumin (Cur) and Chrysin (Chr), were co-encapsulated in PEGylated PLGA NPs and investigated their synergistic inhibitory effect against Caco-2 cancer cells. Characterization of nanoformulated drugs was determined using DLS, FTIR, TEM, and SEM. Drug release study was performed using dialysis method. MTT and real-time PCR assays were applied to evaluate the cytotoxic effects of free and nano-encapsulated drugs on expression level of hTERT in Caco-2 cells. The results showed that free drugs and nano-formulations exhibited a dose-dependent cytotoxicity against Caco-2 cells and especially, Cur-Chr-PLGA/PEG NPs had more synergistic antiproliferative effect and significantly arrested the growth of cancer cells than the other groups (P < 0.05). Real-time PCR results revealed that Cur, Chr, and combination of Cur-Chr in free and encapsulated forms inhibited hTERT gene expression. Also, it was found that Cur-Chr-PLGA/PEG NPs than free combination forms could further decline hTERT expression in all concentration (P < 0.05). In summary, our study represents the first report of nano-combinational application of the natural herbal substances with a one-step fabricated codelivery system for effective colorectal cancer combinational chemotherapy.

Development of quantum-dot-encapsulated liposome-based optical nanobiosensor for detection of telomerase activity without target amplification.

Reactivation of telomerase, which is observed in more than 85% of all known human tumours, is considered a promising tumour marker for cancer diagnosis. With respect to the biomedical importance of telomerase, we have developed a simple strategy based on liposomal fluorescent signal amplification for highly sensitive optical detection of telomerase activity using liposome-encapsulated cadmium telluride quantum dots. In this strategy, telomerase extracted from A549 cells elongated the biotinylated telomerase substrate primer, which was then immobilized on streptavidine-coated microplate wells. After the hybridization of the telomerase-elongated product with biotinylated capture probe, streptavidin was added to the assembly. In the next step, biotinylated liposome was conjugated with capture probe through streptavidin. Finally, QD-encapsulated liposomes were disrupted by Triton X-100, and the fluorescence intensity of the released QDs was measured to detect telomerase activity. The results showed that the proposed nanobiosensor was able to detect telomerase activity from as few as 10 A549 cells without the enzymatic amplification of telomerase extension products. In short, this method is not only convenient and sensitive, but also has a simple operating protocol and a wide detection range (10-5000 cells). A linear range was observed between 50 and 800 cells with a correlation coefficient of 0.982 and regression equation of y = 0.0444 x + 17.137. The proposed method is economical, more user-friendly, without error-prone PCR, with a wide detection range and simple operating protocol without the requirement for sophisticated equipment. Graphical Abstract Schematic representation of the QD-encapsulated liposome-based strategy to amplify fluorescence signal for optical detection of telomerase activity.

Development of Emu oil-loaded PCL/collagen bioactive nanofibers for proliferation and stemness preservation of human adipose-derived stem cells: possible application in regenerative medicine.

Adipose tissue-derived stem cells (ASCs) are promising candidate in stem cell therapies, and maintaining their stemness potential is vital to achieve effective treatment. Natural-based scaffolds have been recently attracted increasing attention in nanomedicine and drug delivery. In the present study, a polymeric nanofibrous scaffold was developed based on the polycaprolactone/Collagen (PCL/Coll) containing Emu oil as a bioactive material to induce the proliferation of ASCs, while simultaneously preserving the stemness property of those cells. Fabrication of the electrospun Emu oil-loaded PCL/Coll nanofibers was confirmed by using FE-SEM, FTIR, and tensile test. ASCs were seeded on two types of nanofibers (PCL/Coll and Emu oil-loaded PCL/Coll) and their proliferation, cell cycle progression, and stemness gene expressions were evaluated using MTT, propidium iodide staining, and qPCR during 14 days, respectively. The results indicated that ASCs displayed improved adhesion capacity with the higher rates of bioactivity and proliferation on the Emu oil-loaded nanofibers than the other groups. The proliferation capacity of ASCs on Emu oil-loaded PCL/Coll nanofibers was further confirmed by the cell cycle progression analysis. It was also found that Emu oil-loaded nanofibers significantly up-regulated the expression of stemness markers including sox-2, nanog, oct4, klf4, and c-Myc. The results demonstrated that the nanofibers containing Emu oil can reinforce the cell adhesion and enhance ASCs proliferation while preserving their stemness; therefore, using scaffolds containing natural products may have a great potential to enhance the in vitro expansion capacity of ASCs in the field of stem cell therapy and regenerative medicine.

Antioxidant effects of chrysin-loaded electrospun nanofibrous mats on proliferation and stemness preservation of human adipose-derived stem cells.

An ideal biomaterial in regenerative medicine should be able to regulate the stem cell proliferation without the loss of its pluripotency. Chrysin (Chr) is a naturally occurring flavone with a wide spectrum of biological functions including anti-inflammatory and anti-oxidant properties. The present study describes the influence of Chr-loaded nanofibrous mats on the regulation of proliferation and stemness preservation of adipose-derived stem cells (ADSCs). For this purpose, Chr-loaded poly (ε-caprolactone)/poly (ethylene glycol) (PCL/PEG) nanofibrous mats were produced via electrospinning process and the successful fabrication of these bioactive mats was confirmed by field emission scanning electron microscopy (FE-SEM) and fourier transform infrared spectroscopy. ADSCs were seeded on the nanofibers and their morphology, viability, and stemness expression were analyzed using FE-SEM, MTT, and qPCR assays after 2 weeks of incubation, respectively. The results display that ADSCs exhibit better adhesion and significantly increased viability on the Chr-loaded PCL/PEG nanofibrous mats in relative to the PCL/PEG nanofibers and tissue culture polystyrene. The greater viability of ADSCs on Chr based nanofibers was further confirmed by higher expression levels of stemness markers Sox-2, Nanog, Oct-4, and Rex-1. These findings demonstrate that Chr-loaded PCL/PEG electrospun nanofibrous mats can be applied to improve cell adhesion and proliferation while concurrently preserving the stemness of ADSCs, thus representing a hopeful potential for application in stem cell therapy strategies.

Cyclodextrin based natural nanostructured carbohydrate polymers as effective non-viral siRNA delivery systems for cancer gene therapy.

Short interfering RNAs (siRNAs), as small non-coding RNA fragments, are one of the widely studied RNAi inducers for gene modulations. The reasonably designed siRNA probes provide a novel potential therapeutic strategy for cancer therapy via silencing the specific cancer-promoting gene. The optimization of physicochemical properties of delivery vectors, such as stability, the possibility of surface functionalization, size, charge, biocompatibility, biodegradability, and non-immunogenicity with receptor-mediated targeting ligands, is necessary for effective intracellular siRNA delivery. The present review is focused on the recent progress of the non-viral nanocarriers for siRNA cancer treatment based on synthetic approaches associated with cyclodextrin (CD)-based carbohydrate polymers, i.e. CD-cationic polymers, CD-polyrotaxanes, CD-dendrimers, and CD-modified tumor-specific targeting ligands. Besides, the efficiency of nanocarriers-based stimuli-responsive CDs is described for the simultaneous delivery of siRNAs and chemotherapeutic drugs. Further, theranostic CD compounds are introduced for the specific diagnosis and cargo-targeting delivery to the specific disease sites. In the meantime, the development of the inherent fluorescent CD-based supramolecular biomaterials without formal chromophores will open up a new strategy to design an effective theranostic non-viral carrier system.

Biomedical applications of zeolite-based materials: A review.

Zeolites are crystalline, hydrated aluminosilicates of alkali earth cations, consisting of 3D frameworks of [SiO4]4- and [AlO4]5- tetrahedral, linked through the shared oxygen atoms, which have been widely applied in multifarious technological approaches such as adsorbents, catalysts, ion exchangers, molecular sieves for separation, and sorting the molecules according to their crystalline size dimensions. On the other hand, the unique and outstanding physical and chemical properties of zeolite materials such as porous character, ion exchangeability, water absorption capacity, immunomodulatory and antioxidative effects, biocompatibility and long-term chemical and biological stability, make them increasingly useful in various filed of biomedicine including drug delivery systems, wound healing, scaffolds used in tissue engineering, anti-bacterial and anti-microbial, implant coating, contrast agents, harmful ions removal from the body, gas absorber, hemodialysis, and teeth root filling. Therefore, this review focuses on the more recent advances of the use of zeolites in various biomedical applications feedbacks especially drug delivery, regenerative medicine, and tissue engineering with special emphasis on their biomaterial perspectives.

Macrophage repolarization using emu oil-based electrospun nanofibers: possible application in regenerative medicine.

In the regenerative medicine therapies, the availability of engineered scaffolds that modulate inflammatory states is highly required. The aim of this study was to evaluate the efficiency of electrospun nanofibrous scaffolds containing natural substances with anti-inflammatory properties such as Emu oil (EO) to control inflammation and re-polarization of macrophages toward M2 anti-inflammatory phonotype. For this purpose, bead free and smooth EO-blended PCL/PEG electrospun nanofibrous mats were successfully fabricated and characterized using FE-SEM, FTIR, and Universal Testing Machine. GC/MS findings of pure EO revealed the fatty acids composition. MTT results showed that macrophage viability on EO-PCL/PEG nanofibres was higher than on PCL/PEG nanofibres and control (p ≤ .05). Additionally, the presence of EO into nanofibres was found to influence on macrophage morphologies, using FE-SEM. qPCR results showed a reduction in iNOS-2 and an increase in Arg-1 levels of macrophages seeded on EO-PCL/PEG nanofibres, indicating the successfully polarization of the macrophages to M2 phenotype. The change in macrophage phenotype on EO-based nanofibres could suppress the inflammation in LPS/IFN-γ stimulated macrophages as evidenced by a major reduction in pro-inflammatory cytokine levels TNF-α, IL-1ß, and IL-6. Conclusively, the results demonstrated that EO-based nanofibres efficiently modulated RAW264.7 macrophage polarity toward an anti-inflammatory M2 phenotype.

Nano-encapsulated metformin-curcumin in PLGA/PEG inhibits synergistically growth and hTERT gene expression in human breast cancer cells.

The study was aimed at investigating the synergistic inhibitory effect of unique combinational regimen of nanocapsulated Metformin (Met) and Curcumin (Cur) against T47D breast cancer cells. For this purpose, Met and Cur were co-encapsulated in PEGylated PLGA nanoparticles (NPs) and evaluated for their therapeutic efficacy. The morphology and dynamic light scattering (DLS) analyses were carried out to optimize the nanoformulations. Drug release study was performed using dialysis method and then the cytotoxic and inhibitory effect of individual and combined drugs on expression level of hTERT in T47D breast cell line were evaluated using MTT assay and qPCR, respectively. The results showed that free drugs and formulations exhibited a dose-dependent cytotoxicity against T47D cells and especially, Met-Cur-PLGA/PEG NPs had more synergistic antiproliferative effect and significantly arrested the growth of cancer cells than the other groups (p < .05). Real-time PCR results revealed that Cur, Met and combination of Met-Cur in free and encapsulated forms inhibited hTERT gene expression. It was found that Met-Cur-PLGA/PEG NPs in relative to free combination could further decline hTERT expression in all concentration (p < .05). Taken together, our study demonstrated that Met-Cur-PLGA/PEG NPs based combinational therapy holds promising potential towards the treatment of breast cancer.

Effects of nano-encapsulated curcumin-chrysin on telomerase, MMPs and TIMPs gene expression in mouse B16F10 melanoma tumour model.

Due to the high rate of drug resistance among malignant melanoma cases, it seems necessary to introduce an efficient pharmaceutical approach to melanoma treatment. For this purpose, Curcumin (Cur) and Chrysin (Chr), two natural anti-cancers, were co-encapsulated in PLGA-PEG nanoparticles (NPs), characterized by DLS, FTIR and FE-SEM and investigated for their effects on MMPs, TIMPs and TERT genes expression in C57B16 mice bearing B16F10 melanoma tumours. The results showed that the expression of MMP-9, MMP-2 and TERT genes were significantly decreased in all treated groups compared to the control. This reduction had the highest amount in CurChr NPs group and then CurChr group for each three genes. Likewise, the expression of TIMP-1 and TIMP-2 genes was significantly increased in all treated groups, compared to the control. Combination groups showed the highest rise in expression of these two genes and the observed increase was greater in nano groups. Moreover, the highest melanoma tumour growth inhibition was detected for CurChr NPs, followed by CurChr = Cur NPs > Cur > Chr NP > Chr. Overall, it is speculated that the nano-combination of Cur and Chr into polymeric NPs with a one-step fabricated co-delivery system may be a promising and convenient approach to improve their efficiency in melanoma cancer therapy.

Chrysin-nanoencapsulated PLGA-PEG for macrophage repolarization: Possible application in tissue regeneration.

The purpose of this study was to investigate the efficiency of a natural flavonoid, Chrysin (Chr), encapsulated in PLGA-PEG nanoparticles (NPs) for the modulation of macrophage polarity from the pro-inflammatory M1 to anti-inflammatory M2 phenotype. The synthetized NPs were characterized using FTIR, DLS and FE-SEM. MTT assay was used to assess the toxicity of different concentration of Chr-encapsulated NPs on LPS/IFN-γ stimulated peritoneal exudate macrophages. To investigate the repolarization efficiency of Chr-encapsulated NPs, real-time PCR was applied to measure M1 (iNOS and SOCS3) and M2 (Arg1 and Fizz) markers expression. Also, the relative mRNA and protein expression levels of pro-inflammatory cytokines including IL-6, IL-1ß and TNF-α were investigated in M1 macrophages treated with Chr-encapsulated NPs. Findings revealed that the Chr-encapsulated NPs with spherical shape and an average diameter of 235 nm were considerably less toxic to the macrophages. Additionally, the nano-formulated Chr efficiently showed a reduction in M1 markers and an increase in M2 markers levels than free Chr. Furthermore, macrophage phenotype switching by PLGA-PEG encapsulated Chr NPs significantly suppressed LPS/IFN-γ induced inflammation by a remarkable reduction in pro-inflammatory cytokine levels, TNF-α, IL-1ß, and IL-6. Convincingly, the results revealed that PLGA-PEG encapsulated Chr based drug delivery system might be introduced into biomaterials to fabricate bioactive smart multifunctional nanocomposites with macrophage repolarization activities for regenerative medicine purposes.

An overview on different strategies for the stemness maintenance of MSCs.

Recent evidence suggests that mesenchymal stem cells (MSCs) have promising therapeutic potential for a broad range of diseases. Because the percentage of MSCs obtained from tissues is very low for cell therapy applications, ex vivo expansion of MSCs is necessary, but aging, loss of stemness and undesired differentiation of them during in vitro cultivation reduces their effectiveness. For achieving ideal therapeutic potential of MSCs in tissue regenerative purposes, it is necessary to retain their stemness properties in vitro. This review emphasis on the last updates in preserving the self-renewal capability of stem cells through in vitro expansion with different parameters.

The Effects of Nanoencapsulated Curcumin-Fe3O4 on Proliferation and hTERT Gene Expression in Lung Cancer Cells.

OBJECTIVE: The aim of the study wasto fabricate curcumin-loaded PLGA-PEG-Fe3O4 nanoparticles and comprise the effects of pure curcumin and curcumin-nanomagnetic encapsulated in PLGA-PEG on cell cytotoxicity and hTERT gene expression in A549 lung cancer cell line. BACKGROUND: Lung cancer is the most common cancer in men and one of the four main cancers that occurs in women. Telomerase is active in more than 85% of various cancerous cells such as lung cancer while its activity is very low in normal cells. Strong evidences of antitumor effects of curcumin; such as the activation of apoptosis, inhibition of angiogenesis and prevention of metastasis, have been confirmed. However, extensive clinical application of this relatively efficacious agent in cancer therapy has been limited because of poor aqueous solubility, and consequently, minimal systemic bioavailability. Nanoparticle-based targeted drug delivery approach has the potential for rendering curcumin specifically at the favorite site using an external magnetic field. It can also improve availability and circumvent the pitfalls of poor solubility. METHODS: Curcumin and Fe3O4 were encapsulated inside the PLGA-PEG co-polymer. Then, the curcumin loaded PLGA-PEG-Fe3O4 nanoparticles were characterized using SEM, FTIR and VSM. In the next step, the cytotoxic effect of different concentrations (0-120 µM) of free curcumin and equivalent doses of curcumin-loaded PLGA-PEG-Fe3O4 was assessed using MTT assay at 24-72 hours. Also, gene expression levels of hTERT were measured through Realtime PCR. RESULTS: By encapsulation of curcumin-Fe3O4, cytotoxicity of the drug substantially increased for all concentrations. IC50 of pure curcumin and nano-encapsulated curcumin during 24, 48 and 72 hours was obtained as 50.5, 49.1 and 48.3 µM and 23.7, 13.6 and 7.3 µM, respectively. Moreover, nano-encapsulated curcumin showed time-dependent cytotoxic effect on A549 cell line during 24, 48, 72 hours in comparison to pure curcumin. In addition, the expression level of the hTERT was reduced with increasing concentrations in both pure and nano-encapsulated curcumin. Compared to pure form, nano-encapsulated curcumin caused further decline in the expression levels of the gene. CONCLUSION: Curcumin incorporating with Fe3O4 loaded into PLGA-PEG co-polymer, as an effective targeted carrier, can make a promising horizon in targeted lung cancer therapy.

Silibinin-loaded magnetic nanoparticles inhibit hTERT gene expression and proliferation of lung cancer cells.

Nanoparticle-based targeted drug delivery has the potential for rendering silibinin specifically at the favorite site using an external magnetic field. Also, it can circumvent the pitfalls of poor solubility. For this purpose, silibinin-loaded magnetic nanoparticles are fabricated, characterized and evaluated cytotoxicity and hTERT gene expression in A549 lung cancer cell line. silibinin-loaded PLGA-PEG-Fe3O4 had dose- and time-dependent cytotoxicity than pure silibinin. Additionally, hTERT expression is more efficiently reduced with increasing concentrations of nanosilibinin than pure silibinin. The present study indicates that PLGA-PEG-Fe3O4 nanoparticles, as an effective targeted carrier, can make a promising horizon in targeted lung cancer therapy.