ROS scavenging activity of polydopamine nanoparticle-loaded supramolecular gelatin-based hydrogel promoted cardiomyocyte proliferation.

Reactive oxygen species (ROS) play essential roles in cellular functions, but maintaining ROS balance is crucial for effective therapeutic interventions, especially during cell therapy. In this study, we synthesized an injectable gelatin-based hydrogel, in which polydopamine nanoparticles were entrapped using supramolecular interactions. The surfaces of the nanoparticles were modified using adamantane, enabling their interactions with ß-cyclodextrin-conjugated with gelatin. We evaluated the cytotoxicity and antioxidant properties of the hydrogel on neonatal rat cardiomyocytes (NRCM), where it demonstrated the ability to increase the metabolic activity of NRCMs exposed to hydrogen peroxide (H2O2) after 5 days. Hydrogel-entrapped nanoparticle exhibited a high scavenging capability against hydroxyl radical, 1'-diphenyl-2-picrylhydrazyl radicals, and H2O2, surpassing the effectiveness of ascorbic acid solution. Notably, the presence of polydopamine nanoparticles within the hydrogel promoted the proliferation activity of NRCMs, even in the absence of excessive ROS due to H2O2 treatment. Additionally, when the hydrogel with nanoparticles was injected into an air pouch model, it reduced inflammation and infiltration of immune cells. Notably, the levels of anti-inflammatory factors, IL-10 and IL-4, were significantly increased, while the pro-inflammatory factor TNF-α was suppressed. Therefore, this novel ROS-scavenging hydrogel holds promise for both efficient cell delivery into inflamed tissue and promoting tissue repair.

Conditioned media from human pluripotent stem cell-derived cardiomyocytes inhibit the growth and migration of lung cancer cells.

Conditioned media (CM) from various cell types contain significant levels of paracrine factors. Recently, therapeutic properties of CM derived from stem cells have been revealed. Based on the fact that heart cancer is extremely rarely, we hypothesized that the CM obtained from human pluripotent stem cell-derived cardiomyocytes might inhibit cancer cell growth and survival. To this end, lung cancer cell line A549 along with human foreskin fibroblasts (HFF) were treated with serial concentrations of cardiomyocyte CM (CCM) or fibroblast CM (FCM). We found that CCM markedly reduced the viability of lung cancer cells, while FCM did not compromise the viability of neither cancer cells nor HFF cells. Furthermore, we determined an optimized CCM concentration, 30 mg/mL, at which the growth, clonogenicity, and migration of A549 and Calu6 lung cancer cell lines were substantially impaired, whereas FCM did not influence these properties. Moreover, lung cancer cells exhibited cell cycle regulation upon treatment with CCM and the rate of apoptosis was markedly increased by cardiomyocyte CM in both lung cancer cell lines tested. Finally, in response to CCM treatment, A549 and Calu6 cells expressed lower levels of antiapoptotic and stemness genes, but higher levels of proapoptotic genes. In conclusion, this study provides cellular and molecular evidence for the antitumor ability of secretome obtained from stem cell-derived cardiomyocytes.

PD-L1-Targeted Co-Delivery of Two Chemotherapeutics for Efficient Suppression of Skin Cancer Growth.

To overcome the severe side effects of cancer chemotherapy, it is vital to develop targeting chemotherapeutic delivery systems with the potent inhibition of tumour growth, angiogenesis, invasion and migration at low drug dosages. For this purpose, we co-loaded a conventional antiworm drug, albendazole (ABZ), and a TOPK inhibitor, OTS964, into lipid-coated calcium phosphate (LCP) nanoparticles for skin cancer treatment. OTS- and ABZ-loaded LCP (OTS-ABZ-LCP) showed a synergistic cytotoxicity against skin cancer cells through their specific cancerous pathways, without obvious toxicity to healthy cell lines. Moreover, dual-targeting the programmed death ligand-1 (PD-L1) and folate receptor overexpressed on the surface of skin cancer cells completely suppressed the skin tumour growth at low doses of ABZ and OTS. In summary, ABZ and OTS co-loaded dual-targeting LCP NPs represent a promising platform with high potentials against complicated cancers where PD-L1/FA dual targeting appears as an effective approach for efficient and selective cancer therapy.

The conserved long non-coding RNA CARMA regulates cardiomyocyte differentiation.

AIMS: Production of functional cardiomyocytes from pluripotent stem cells requires tight control of the differentiation process. Long non-coding RNAs (lncRNAs) exert critical regulatory functions in cell specification during development. In this study, we designed an integrated approach to identify lncRNAs implicated in cardiogenesis in differentiating human embryonic stem cells (ESCs). METHODS AND RESULTS: We identified CARMA (CARdiomyocyte Maturation-Associated lncRNA), a conserved lncRNA controlling cardiomyocyte differentiation and maturation in human ESCs. CARMA is located adjacent to MIR-1-1HG, the host gene for two cardiogenic miRNAs: MIR1-1 and MIR-133a2, and transcribed in an antisense orientation. The expression of CARMA and the miRNAs are negatively correlated, and CARMA knockdown increases MIR1-1 and MIR-133a2 expression. In addition, CARMA possesses MIR-133a2 binding sites, suggesting the lncRNA could be also a target of miRNA action. Upon CARMA down-regulation, MIR-133a2 target protein-coding genes are coordinately down-regulated. Among those, we found RBPJ, the gene encoding the effector of the NOTCH pathway. NOTCH has been shown to control a binary cell fate decision between the mesoderm and the neuroectoderm lineages, and NOTCH inhibition leads to enhanced cardiomyocyte differentiation at the expense of neuroectodermal derivatives. Interestingly, two lncRNAs, linc1230 and linc1335, which are known repressors of neuroectodermal specification, were found up-regulated upon Notch1 silencing in ESCs. Forced expression of either linc1230 or linc1335 improved ESC-derived cardiomyocyte production. These two lncRNAs were also found up-regulated following CARMA knockdown in ESCs. CONCLUSIONS: Altogether, these data suggest the existence of a network, implicating three newly identified lncRNAs, the two myomirs MIR1-1 and MIR-133a2 and the NOTCH signalling pathway, for the coordinated regulation of cardiogenic differentiation in ESCs.

ATP stabilised and sensitised calcium phosphate nanoparticles as effective adjuvants for a DNA vaccine against cancer.

Cancer vaccines based on DNA encoding oncogenes have shown great potential in preclinical studies. However, the efficacy of DNA vaccines is limited by their weak immunogenicity because of low cellular internalisation and insufficient activation of dendritic cells (DCs). Calcium phosphate (CP) nanoparticles (NPs) are biodegradable vehicles with low toxicity and high loading capacity of DNA but suffer from stability issues. Here we employed adenosine triphosphate (ATP) as a dual functional agent, i.e. stabiliser for CP and immunological adjuvant, and applied the ATP-modified CP (ACP) NPs to the DNA vaccine. ACP NP-enhanced cellular uptake and improved transfection efficiency of DNA vaccine, and further showed the ability to activate DCs that are critical for them to prime T cells in cancer immunotherapy. As a result, a higher level of antigen-specific antibody with stronger tumour growth inhibition was achieved in mice immunised with the ACP-DNA vaccine. Overall, this one-step synthesised ACP NPs are an efficient nano-delivery system and nano-adjuvant for cancer DNA vaccines.

Mannose-Functionalized Biodegradable Nanoparticles Efficiently Deliver DNA Vaccine and Promote Anti-tumor Immunity.

Cancer vaccines have attracted increasing attention for their application in tumor immunotherapy. DNA vaccines are one of them that have been proven very promising with the advantages of safety, rapid design, and low cost. However, the low stability, ineffective cell internalization, and low immunostimulation hinder their wide application. Thus, developing targeted and safe systems to effectively deliver DNA vaccines becomes a vital step. In this study, we report the development of mannose- and bisphosphonate (BP)-modified calcium phosphate (CP) nanoparticles (NPs) as efficient vaccine delivery vehicles by targeting C-type lectin receptors (CLRs) on antigen-presenting cells (APCs). Using a model antigen ovalbumin (OVA)-encoded plasmid DNA (pOVA) as a model vaccine, we demonstrate that mannose-modified and BP-stabilized CP (MBCP) nanoparticles are mono-dispersed for enhanced uptake by APCs and subsequently induce OVA antigen presentation and immunostimulation. Mice immunized with MBCP-pOVA nanovaccines show a significantly stronger anti-OVA antibody response with a quicker IgG1 and IgG2a antibody production than unmodified NPs. Moreover, MBCP-pOVA immunization significantly inhibits the growth of OVA-expressing E.G7 tumor cells in C57BL/6J mice. Our data collectively suggest that the modifications to enhance the stability and targeting ability of MBCP NPs are essential for effective delivery of DNA vaccines and promote robust anti-tumor immunity.

Enhanced Prevention of Breast Tumor Metastasis by Nanoparticle-Delivered Vitamin E in Combination with Interferon-Gamma.

Preventing cancer metastasis is one of the remaining challenges in cancer therapy. As an efficient natural product, alpha-tocopheryl succinate (α-TOS), the most effective form of vitamin E, holds great anticancer potential. To improve its efficacy and bioavailability, lipid-coated calcium carbonate/phosphate (LCCP) nanoparticles (NPs) with folic acid and PEG modification are synthesized for efficient delivery of α-TOS to 4T1 cancer cells. The optimized LCCP-FA NPs (NP-TOS15) show an α-TOS loading efficiency of around 60%, and enhanced uptake by 4T1 metastatic cancer cells. Consequently, NP-TOS15 significantly enhance the anticancer effect in combination with interferon-gamma (IFN-γ) in terms of apoptosis facilitation and migration inhibition. Importantly, NP-TOS15 upregulate the anticancer immunity via downregulating program death ligand 1 (PD-L1) expression that is initially induced by IFN-γ, and remarkably prevent the lung metastasis, particularly in combination with IFN-γ. Further investigation reveals that this combination therapy also modulates the cytotoxic lymphocyte infiltration into the tumor microenvironment for tumor elimination. Taken together, the NP delivery of α-TOS in combination with IFN-γ provides an applicable strategy for cancer therapy.

Nanostructuring a Widely Used Antiworm Drug into the Lipid-Coated Calcium Phosphate Matrix for Enhanced Skin Tumor Treatment.

Recently, a widely used antiworm drug albendazole (ABZ) has been recognized as an anticancer drug, while being safe for healthy cells. However, its clinical application is limited due to low water solubility. In this work, we incorporated ABZ into pH-responsive lipid-coated calcium phosphate (LCP) nanoparticles to enhance anticancer efficacy. LCP-ABZ nanoparticles doubled the ABZ solubility with a much quicker release rate in the slightly acidic environment. LCP-ABZ significantly improved cytotoxicity to B16F0 cells and reduced their migration probably through enhanced apoptosis induction via reactive oxygen species overproduction. In a B16F0-bearing mouse models, LCP-ABZ nanoparticles reduced the tumor size by 50-60% at the total dose of only 15 mg (ABZ)/kg, more effective than commercial albendazole and other ABZ nanoparticles. Overall, this research demonstrated LCP nanoparticle as an effective platform to deliver ABZ for cancer therapy, with safety to healthy tissues and the potential to inhibit metastasis.

Nanoformulations of albendazole as effective anticancer and antiparasite agents.

Initially emerging as a widely used clinical antiparasitic drug, albendazole (ABZ) has been increasingly recognized as an effective anticancer agent due to its outstanding advantage, in other words, low toxicity to normal cells but high effectiveness against parasites and some tumors. The major challenge is its poor water solubility and subsequently low bioavailability. This article thus first reviews the brief achievements in using ABZ to treat parasites and cancers, and summarizes the basic mechanisms of action of ABZ. Then this article critically reviews recent nanotechnological strategies, in other words, formulating/conjugating it with carriers into nanoformulations, in practices of improving aqueous solubility and efficacy in treatment of tumors and parasites. Our expert opinions in this field are provided for more effective delivery of ABZ to treat tumors and parasites in vivo.

Near-infrared light-sensitive liposomes for enhanced plasmid DNA transfection.

Near-infrared (NIR) light-responsive liposomes are attractive carriers for targeted and controlled drug delivery to the superficial organ or tissue (e.g., skin). This work describes the development of NIR-responsive liposomes by incorporating gold nanostars within liposomes composed of Phospholipon 90 g and cholesterol. Following cellular delivery, photothermal effect around the gold nanostar upon NIR stimulation induces microcavitation and liposome phase transition which consequently triggers the release of encapsulated molecules. Taking GFP plasmid as an example, we demonstrate enhanced gene transfection into fibroblasts following NIR treatment.

Stimuli-responsive liposomes for the delivery of nucleic acid therapeutics.

Nucleic acid therapeutics (NATs) are valuable tools in the modulation of gene expression in a highly specific manner. So far, NATs have been actively pursued in both pre-clinical and clinical studies to treat diseases such as cancer, infectious and inflammatory diseases. However, the clinical application of NATs remains a considerable challenge owing to their limited cellular uptake, low biological stability, off-target effect, and unfavorable pharmacokinetics. One concept to address these issues is to deliver NATs within stimuli-responsive liposomes, which release their contents of NATs upon encountering environmental changes such as temperature, pH, and ion strength. In this case, before reaching the targeted tissue/organ, NATs are protected from degradation by enzymes and immune system. Once at the area of interest, localized and targeted delivery can be achieved with minimal influence to other parts of the body. Here, we discuss the latest developments and existing challenges in this field. FROM THE CLINICAL EDITOR: Nucleic acid therapeutics have been shown to enhance or eliminate specific gene expression in experimental research. Unfortunately, clinical applications have so far not been realized due to problems of easy degradation and possible toxicity. The use of nanosized carriers such as liposomes to deliver nucleic acids is one solution to overcome these problems. In this review article the authors describe and discuss the potentials of various trigger-responsive "smart" liposomes, with a view to help other researchers to design better liposomal nucleic acid delivery systems.

Effect of gold nanoparticles on properties of nanoliposomal hydroxyurea: an in vitro study.

New hopes in cancer treatment have been emerged using functional nanoparticles. In this work, we tried to synthesize gold nanoparticles and gold nanoparticles conjugated with DNA extracted from human breast cancer cells. After synthesizing, gold nanoparticles were mixed with nanoliposomal hydroxyurea and corresponding compounds were formed. They were described by UV-Visible spectrophotometry and Zeta sizer. Amount of drug loading into liposomes was determined by spectrophotometry and cytotoxicity effect on MCF-7 cells was measure by MTT assay. Drug loading was determined to be 70 %. Size, size distribution and Zeta potential of particles were 473 nm, 0.46 and -21 mV for control nanoliposomal ones and 351 nm, 0.38 and -25 mV for nanoliposomal particles containing hydroxyurea. This was 29 nm, 0.23 and -30 mV for gold nanoparticles and 502 nm, 0.41 and -38 mV for nanoliposomes containing drug loaded by gold nanoparticles conjugated with DNA. It was found that nano conjugated complex in concentrations less than 20 µM of hydroxyurea can improve efficiency compared with liposomal drug. In maximum concentration of drug (2,500 µM), cytotoxicity was equal to 95 %. In minimum concentration of drug (5 µM), cytotoxicity of liposomal drug and conjugated complex were 70 and 81 %, respectively which probably comes from increased drug entry into cells due to the presence of gold nanoparticles. Free drug resulted in toxicity of 32 % in 5 µM and 88 % in 2,500 µM. Results demonstrated higher drug efficiency in nanoparticle form compared with free form which can be used in in vivo studies.

Polybutylcyanoacrylate nanoparticles and drugs of the platinum family: last status.

Cisplatinum and carboplatinum drugs from platinum-containing family are anti-cancer drugs. Using these drugs causes side effects. Targeted and selective prescription decreases side effects of the drugs. This can be achieved using nanotechnology. In this study, cisplatinum and carboplatinum drugs were loaded on polybutylcyanoacrylate nanoparticles using emulsion polymerization method. To determine amount of loaded drug onto nanoparticle, spectrophotometry method was used. Evaluation of cytotoxicity of such nanoparticles was performed on MCF-7 cell line using MTT assay. Loading percentage of cisplatinum and carboplatinum drugs on nanoparticles were estimated 4 and 6 %, respectively. Cytotoxicity survival rate for cisplatinum and nanoparticle containing cisplatinum at the lowest concentration (p < 0.01) (20 µM) were estimated 64 ± 1 and 67 ± 0.5 %, respectively. These values at the highest concentration (p < 0.01) (160 µM) were measured 28 ± 0.7 and 31 ± 0.4 %. Additionally for carboplatinum and nanoparticles containing carboplatinum at the concentration (p < 0.01) (20 µM) amounts were estimated to be 80 ± 0.6 and 84 ± 0.6 %, while at the concentration (p < 0.01) (160 µM) were identified to be 44 ± 0.5 and 51 ± 0.2 %, respectively. Probably, due to low level of loading, cytotoxicity of both drugs at nano particle status was decreased in comparison with their standard form.

Release modeling and comparison of nanoarchaeosomal, nanoliposomal and pegylated nanoliposomal carriers for paclitaxel.

Breast cancer is the most prevalent cancer among women. Recently, delivering by nanocarriers has resulted in a remarkable evolution in treatment of numerous cancers. Lipid nanocarriers are important ones while liposomes and archaeosomes are common lipid nanocarriers. In this work, paclitaxel was used and characterized in nanoliposomal and nanoarchaeosomal form to improve efficiency. To increase stability, efficiency and solubility, polyethylene glycol 2000 (PEG 2000) was added to some samples. MTT assay confirmed effectiveness of nanocarriers on MCF-7 cell line and size measuring validated nano-scale of particles. Nanoarchaeosomal carriers demonstrated highest encapsulation efficiency and lowest release rate. On the other hand, pegylated nanoliposomal carrier showed higher loading efficiency and less release compared with nanoliposomal carrier which verifies effect of PEG on improvement of stability and efficiency. Additionally, release pattern was modeled using artificial neural network (ANN) and genetic algorithm (GA). Using ANN modeling for release prediction, resulted in R values of 0.976, 0.989 and 0.999 for nanoliposomal, pegylated nanoliposomal and nanoarchaeosomal paclitaxel and GA modeling led to values of 0.954, 0.951 and 0.976, respectively. ANN modeling was more successful in predicting release compared with the GA strategy.

Archaeosome: as new drug carrier for delivery of Paclitaxel to breast cancer.

In the present study, paclitaxel was archaeosomed to reduce side effects and improve its therapeutic index. Carriers have made a big evolution in treatment of many diseases in recent years. Lipid carriers are of special importance among carriers. Archaeosome is one of the lipid carriers. Paclitaxel is one of the drugs used to treat breast cancer which has some unwanted side effects despite its therapeutic effects. Archaeosomes were extracted from methanogenic archi bacteria and synthesized with a certain ratio of paclitaxel in PBS. The mean diameter of archaeosomal paclitaxel was measured by Zeta sizer instrument, Drug releasing of archaeosomal paclitaxel was examined within 26 h which results showed that the most drug releasing occurs during first 3 h. The cytotoxicity effect of archaeosomal paclitaxel on breast cancer's cell line was evaluated by MTT assay which results showed that the cytotoxicity effect of archaeosomal paclitaxel on breast cancer's cell line is more than that of the standard paclitaxel formulation. The results indicated that new drug delivery of paclitaxel using archaeosome, increases the therapeutic index of the drug.

Effect of sodium chloride on efficiency of cisplatinum dissolved in dimethyl sulfoxide: an in vitro study.

Cisplatinum (Cispt) is an anti-cancer drug with a low level of solubility. One of Cispt's solvents is dimethyl sulfoxide (DMSO) which can be substituted with chlorine of drug as Cispt's solvent. Applying such a solvent in biological studies is impossible due to intense reduction in activity. On the other hand, it is specified that Cispt's stability is increased in aqueous media by increasing sodium chloride (NaCl) concentration up to 0.9 %. Consequently, we intended to study the effect of DMSO on cytotoxicity of Cispt in presence of sodium. MTT assay was employed to study cytotoxicity effect of Cispt + NaCl + DMSO and Cispt + DMSO on G-292 cell line. Cytotoxicity in dilutions of 300 and 9 (p < 0.01) of Cispt in Cispt + NaCl + DMSO formulation was equal to 78 and 7 %. These values were estimated 79 and 18 % for Cispt + DMSO formulation and 79 and 24 % for free drug. IC50 values demonstrated reduction of 45 % in cytotoxicity of Cispt in Cispt + DMSO formulation. Studying chemical structure of Cispt and Cispt dissolved in DMSO showed that NaCl cannot inhibit inactivating effect of DMSO on Cispt and effect of this solvent on Cispt is independent from presence of NaCl. Results represented that using NaCl does not result in stability and keeping cytotoxicity properties of Cispt in DMSO. Findings suggest more studies for using DMSO as a solvent of Cispt.

Efficacy of Cisplatin-loaded polybutyl cyanoacrylate nanoparticles on the glioblastoma.

Glioblastoma is known as one of the most aggressive human cancers. To gain access of the brain, therapeutic agents must overcome blood-brain barrier (BBB). In this study, Cisplatin (Cispt)-loaded polybutylcyanoacrylate (PBCA) nanoparticles (NPs) were prepared through miniemulsion polymerization technique. They were coated with polysorbate 80 to cross the BBB of glioblastoma-bearing rats. Prepared NPs were characterized with respect to their size, size distribution, zeta potential, drug loading and encapsulation efficiency, cytotoxicity effects, drug release, and stability pattern. Size and zeta potential of nanodrug were found to be 489 nm and -20 mV, while drug loading and encapsulation efficiency were determined to be 5% and 25%, respectively. Release studies demonstrated high retention capability of nanodrug in that 3.18% of Cispt was released from NPs in a period of 51 h. NPs presented acceptable stability after 2 months and lyophilization. Mean survival time in nanodrug receivers was 19.6 days, while it was 17.5 days for free drug receivers. Histological studies demonstrated efficacy of PBCA NPs in reducing side effects. Finally, such preparation can be considered as a promising nanocarrier for other types of tumor.

Drug delivery of hydroxyurea to breast cancer using liposomes.

It is clear that cancer is one of the most mortal diseases in the world and the most prevalent among women is breast cancer. As hydroxyurea (HU)-a drug which is used in chemotherapy-has many adverse effects in long-term despite of its therapeutic properties, we made use of nano drug delivery technology in order to reduce adverse effects and increase therapeutic index. Thus, liposomation is a novel way in drug delivery systems. In this study a mixture of phosphatidylcholine and cholesterol was mixed and HU was added to the resultant mixture. The mean diameter of the nanoliposomal HU measured with the Zeta Sizer device (equal to 402.5 nm) and its encapsulation efficiency was 70.8 %. Besides, using dialysis, the pattern of drug release from nanoliposomes has been studied and the results showed that the drug release of nanoliposomal drug within 28 h was equal to 25.85 %. This study showed that the cytotoxicity effect of nanoliposomal drug is more than that of the standard drug.

Cytotoxicity of liposomal Paclitaxel in breast cancer cell line mcf-7.

Regarding that the breast cancer is the most prevalent disease among women, paclitaxel, an anti-cancer drug, could be used in treatment of this disease. As paclitaxel has adverse effects, it was used of nanoliposome drug delivery technology in order to reduce adverse effects and improve drug efficacy. Certain ratios of phosphatidylcholine, cholesterol and paclitaxel were synthesized to prepare nanoliposomal paclitaxel. Using Zeta sizer device, the mean diameter of nanoliposomal paclitaxel was obtained 421.4 nm and its encapsulation efficiency was 91.3 %. By dialysis, drug release in nanoliposome paclitaxel formulation within 28 h was studied which was 5.53 %. This study showed that cytotoxicity effect of nanoliposomal paclitaxel is more than that of the standard form.