Nanogels with covalently bound and releasable trehalose for autophagy stimulation in atherosclerosis.

Atherosclerosis, cholesterol-driven plaque formation in arteries, is a complex multicellular disease which is a leading cause of vascular diseases. During the progression of atherosclerosis, the autophagic function is impaired, resulting in lipid accumulation-mediated foam cell formation. The stimulation of autophagy is crucial for the recovery of cellular recycling process. One of the potential autophagy inducers is trehalose, a naturally occurring non-reducing disaccharide. However, trehalose has poor bioavailability due to its hydrophilic nature which results in poor penetration through cell membranes. To enhance its bioavailability, we developed trehalose-releasing nanogels (TNG) for the treatment of atherosclerosis. The nanogels were fabricated through copolymerization of 6-O-acryloyl-trehalose with the selected acrylamide-type monomers affording a high trehalose conjugation (~ 58%, w/w). TNG showed a relatively small hydrodynamic diameter (dH, 67 nm) and a uniform spherical shape and were characterized by negative ζ potential (-18 mV). Thanks to the trehalose-rich content, TNG demonstrated excellent colloidal stability in biological media containing serum and were non-hemolytic to red blood cells. In vitro study confirmed that TNG could stimulate autophagy in foam cells and enhance lipid efflux and in vivo study in ApoE-/- mice indicated a significant reduction in atherosclerotic plaques, while increasing autophagic markers. In conclusion, TNG hold great promise as a trehalose delivery system to restore impaired autophagy-mediated lipid efflux in atherosclerosis and subsequently reduce atherosclerotic plaques.

Trehalose-Bearing Carriers to Target Impaired Autophagy and Protein Aggregation Diseases.

In recent years, trehalose, a natural disaccharide, has attracted growing attention because of the discovery of its potential to induce autophagy. Trehalose has also been demonstrated to preserve the protein's structural integrity and to limit the aggregation of pathologically misfolded proteins. Both of these properties have made trehalose a promising therapeutic candidate to target autophagy-related disorders and protein aggregation diseases. Unfortunately, trehalose has poor bioavailability due to its hydrophilic nature and susceptibility to enzymatic degradation. Recently, trehalose-bearing carriers, in which trehalose is incorporated either by chemical conjugation or physical entrapment, have emerged as an alternative option to free trehalose to improve its efficacy, particularly for the treatment of neurodegenerative diseases, atherosclerosis, nonalcoholic fatty liver disease (NAFLD), and cancers. In the current Perspective, we discuss all existing literature in this emerging field and try to identify key challenges for researchers intending to develop trehalose-bearing carriers to stimulate autophagy or inhibit protein aggregation.

A Simple Synthesis of Reduction-Responsive Acrylamide-Type Nanogels for miRNA Delivery.

MicroRNAs (miRNAs) have great therapeutic potential; however, their delivery still faces huge challenges, especially given the short half-life of naked miRNAs due to rapid hydrolysis or inactivation by abundant nucleases in the systemic circulation. Therefore, the search for reliable miRNA delivery systems is crucial. Nanogels are one of the more effective nanocarriers because they are biocompatible and have a high drug-loading capacity. In this study, acrylamide-based nanogels containing cationic groups and redox-sensitive crosslinkers were developed for cellular delivery of anti-miR21 (a-miR21). To achieve this, post-polymerization loading of a-miR21 oligonucleotides into nanogels was performed by utilizing the electrostatic interaction between positively charged nanogels and negatively charged oligonucleotides. Different molar ratios of the amine groups (N) on the cationic nanogel and phosphate groups (P) on the miRNA were investigated. An N/P ratio of 2 allowed high miRNA loading capacity (MLC, 6.7% w/w) and miRNA loading efficiency (MLE, 99.7% w/w). Successful miRNA loading was confirmed by dynamic light scattering (DLS) and electrophoretic light scattering (ELS) measurements. miRNA-loaded nanogels (NG/miRNA) formed stable dispersions in biological media and showed an enhanced miRNA release profile in the presence of glutathione (GSH). Moreover, the addition of heparin to dissociate the miRNA from the cationic nanogels resulted in the complete release of miRNA. Lastly, a cell uptake study indicated that NG/miRNA could be easily taken up by cancer cells.

White saffron (Curcuma mangga Val.) attenuates diabetes and improves pancreatic ß-cell regeneration in streptozotocin-induced diabetic rats.

Diabetes is a chronic disease caused by an imbalance of insulin release to the bloodstream in response to excessive glucose influx, which causes hyperglycemia. White saffron (Curcuma mangga Val.), an Indonesian aromatic spice, contains essential phytochemicals and has a number of potential health benefits. Here, we examined the effects of oral administration of white saffron powder (WSP): 1.5 and 4.5 g (WSP 1.5 and WSP 4.5) in streptozotocin (STZ)-induced diabetic rats. WSP was administered orally on a daily basis for one month and its antidiabetic, anti-inflammatory, and antioxidative effects were investigated by measuring the concentrations of blood glucose, insulin, interleukin 6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin 8 (IL-8), malondialdehyde (MDA), and superoxide dismutase (SOD) activity. In response to high WSP intervention (WSP 4.5), treated rats showed increased insulin level and SOD activity and reduced blood glucose, IL-6, IL-8, TNF-α, and MDA levels, which were closely related to the positive control (PC) group. In addition, Hematoxylin and Eosin (H&E) staining of the pancreatic tissues showed that WSP 4.5-treated rats had significant improvement in ß-cell regeneration, which taken together reflected the antidiabetic potential of Curcuma mangga Val.

pH and Reduction Dual-Responsive Nanogels as Smart Nanocarriers to Resist Doxorubicin Aggregation.

The use of smart nanocarriers that can modulate therapeutic release aided by biological cues can prevent undesirable cytotoxicity caused by the premature release of cytotoxic drugs during nanocarrier circulation. In this report, degradable nanocarriers based on pH/reduction dual-responsive nanogels were synthesized to encapsulate doxorubicin hydrochloride (DOX) and specifically boost the release of DOX in conditions characteristic of the cancer microenvironment. Nanogels containing anionic monomer 2-carboxyethyl acrylate (CEA) and N,N'-bis(acryloyl)cystamine (CBA) as a degradable crosslinker have been successfully synthesized via photoinitiated free radical polymerization. The loading process was conducted after polymerization by taking advantage of the electrostatic interaction between the negatively charged nanogels and the positively charged DOX. In this case, a high drug loading capacity (DLC) of up to 27.89% was achieved. The entrapment of DOX into a nanogel network could prevent DOX from aggregating in biological media at DOX concentrations up to ~160 µg/mL. Anionic nanogels had an average hydrodynamic diameter (dH) of around 90 nm with a negative zeta (ζ) potential of around -25 mV, making them suitable for targeting cancer tissue via the enhanced permeation effect. DOX-loaded nanogels formed a stable dispersion in different biological media, including serum-enriched cell media. In the presence of glutathione (GSH) and reduced pH, drug release was enhanced, which proves dual responsivity. An in vitro study using the HCT 116 colon cancer cell line demonstrated the enhanced cytotoxic effect of the NG-CBA/DOX-1 nanogel compared to free DOX. Taken together, pH/reduction dual-responsive nanogels show promise as drug delivery systems for anticancer therapy.

Trehalose-releasing nanogels: A step toward a trehalose delivery vehicle for autophagy stimulation.

Trehalose has been widely studied as a treatment for a variety of human disorders due to its ability to stimulate autophagy. Trehalose, however, is poorly adsorbed and is hydrolyzed in the intestinal mucosa, and oral delivery requires relatively high doses to induce autophagy. The parenteral injection of trehalose-releasing nanogels proposed in this study offers an alternative mode of delivery. This study aimed to develop stable colloidal dispersions of trehalose-rich nanogels that could sustainably release trehalose under physiologically relevant conditions. The nanogel design was based on the covalent incorporation of 6-O-acryloyl-trehalose within a polymer network. A series of nine trehalose-rich nanogels with highly conjugated trehalose (up to 59 % w/w) were synthesized and shown to sustainably release trehalose at a rate that is not dose dependent. The nanogels were optimized to keep colloidal stability in serum-enriched cell culture media. The stable nanogels were not cytotoxic to primary HUVECs. Two selected nanogels with opposite surface charges were subjected to extended in vitro characterization that included a cellular uptake study and a hemocompatibility assay. Both nanogels were efficiently taken up by HUVECs during a short incubation. They also proved not to be hemolytic to human RBCs in concentrations up to 2.0 mg/mL. Finally, an in vivo autophagy stimulation study employing transgenic zebrafish and Drosophila larvae demonstrated that prolonged exposure to a cationic trehalose-releasing nanogel can induce autophagic activity in in vivo systems without any detectable toxicity.

Macrophage membrane camouflaged reactive oxygen species responsive nanomedicine for efficiently inhibiting the vascular intimal hyperplasia.

BACKGROUND: Intimal hyperplasia caused by vascular injury is an important pathological process of many vascular diseases, especially occlusive vascular disease. In recent years, Nano-drug delivery system has attracted a wide attention as a novel treatment strategy, but there are still some challenges such as high clearance rate and insufficient targeting. RESULTS: In this study, we report a biomimetic ROS-responsive MM@PCM/RAP nanoparticle coated with macrophage membrane. The macrophage membrane with the innate "homing" capacity can superiorly regulate the recruitment of MM@PCM/RAP to inflammatory lesion to enhance target efficacy, and can also disguise MM@PCM/RAP nanoparticle as the autologous cell to avoid clearance by the immune system. In addition, MM@PCM/RAP can effectively improve the solubility of rapamycin and respond to the high concentration level of ROS accumulated in pathological lesion for controlling local cargo release, thereby increasing drug availability and reducing toxic side effects. CONCLUSIONS: Our findings validate that the rational design, biomimetic nanoparticles MM@PCM/RAP, can effectively inhibit the pathological process of intimal injury with excellent biocompatibility.

ROS-responsive biomimetic nanoparticles for potential application in targeted anti-atherosclerosis.

The development of nanomedicines provides new opportunities for the treatment of atherosclerosis (AS) due to their great advantages such as the improved drug solubility, enhanced bioavailability and reduced side effects. Despite these advantages, nanomedicines are still facing some challenges. The problems remain in the short circulation life, lack of specific targeting and poor drug release controllability. In order to overcome the shortages of conventional nanomedicines, the combination of biomimetic strategy with smart nanoagents has been proposed. In light with the high reactive oxygen species (ROS) level in AS microenvironment and the fact that macrophages play a critical role in the pathogenesis of AS, we fabricated ROS-responsive biomimetic nanoparticles (NPs), which camouflaged macrophage membrane (MM) on ROS-responsive NPs loaded with rapamycin (RNPs) for potential application in AS therapy. The resulting ROS-responsive biomimetic NPs (MM/RNPs) exhibited favorable hydrodynamic size with negative surface charge, retained the functional proteins from MM, and showed ROS-responsive drug release. Because of the biomimetic camouflaging on surface, MM/RNPs could effectively escape from macrophages uptake and target to inflammatory endothelial cells. Meanwhile, MM/RNPs could inhibit the proliferation of macrophages and smooth muscle cells in vitro. Furthermore, the MM-coated NPs were found to be nontoxic in both cytotoxicity assay and in vivo toxicity evaluation. Consequently, these results demonstrated that MM/RNPs could be a potential candidate of drug delivery system for safe and effective anti-AS applications.

Engineered bioresponsive nanotherapeutics: recent advances in the treatment of atherosclerosis and ischemic-related disease.

Biological stimuli that are present during the pathogenesis of disease have gained considerable interest as a critical element for the design of smart drug delivery systems. Recently, the utilization of biological stimuli-responsive (bioresponsive) nanotheranostic agents to treat atherosclerosis and ischemic-related diseases has demonstrated significant outcomes in preclinical studies. Those diseases share similar hallmarks, including high levels of endogenous reactive oxygen species (ROS), low pH, and high enzyme activity. Interestingly, other relevant biological stimuli such as shear stress, cholesterol, and glutathione have recently been explored as internal stimuli to trigger drug release and some particular actions. In addition, a number of strategies can be proposed to enhance their targeting efficiency, diagnostic properties, and efficacy rate. This review discusses recent advancements in the preclinical studies of bioresponsive nanotherapeutics as diagnostic and therapeutic agents against atherosclerosis and ischemic-related diseases as well as some potential strategies to overcome the current limitations.

Macrophage membrane functionalized biomimetic nanoparticles for targeted anti-atherosclerosis applications.

Atherosclerosis (AS), the underlying cause of most cardiovascular events, is one of the most common causes of human morbidity and mortality worldwide due to the lack of an efficient strategy for targeted therapy. In this work, we aimed to develop an ideal biomimetic nanoparticle for targeted AS therapy. Methods: Based on macrophage "homing" into atherosclerotic lesions and cell membrane coating nanotechnology, biomimetic nanoparticles (MM/RAPNPs) were fabricated with a macrophage membrane (MM) coating on the surface of rapamycin-loaded poly (lactic-co-glycolic acid) copolymer (PLGA) nanoparticles (RAPNPs). Subsequently, the physical properties of the MM/RAPNPs were characterized. The biocompatibility and biological functions of MM/RAPNPs were determined in vitro. Finally, in AS mouse models, the targeting characteristics, therapeutic efficacy and safety of the MM/RAPNPs were examined. Results: The advanced MM/RAPNPs demonstrated good biocompatibility. Due to the MM coating, the nanoparticles effectively inhibited the phagocytosis by macrophages and targeted activated endothelial cells in vitro. In addition, MM-coated nanoparticles effectively targeted and accumulated in atherosclerotic lesions in vivo. After a 4-week treatment program, MM/RAPNPs were shown to significantly delay the progression of AS. Furthermore, MM/RAPNPs displayed favorable safety performance after long-term administration. Conclusion: These results demonstrate that MM/RAPNPs could efficiently and safely inhibit the progression of AS. These biomimetic nanoparticles may be potential drug delivery systems for safe and effective anti-AS applications.

Hypoglycemic Activity of Curcuma mangga Val. Extract via Modulation of GLUT4 and PPAR-γ mRNA Expression in 3T3-L1 Adipocytes.

BACKGROUND: There would be over 600 million people living with diabetes by 2040 as predicted by the World Health Organization. Diabetes is characterized by raised blood sugar and insulin resistance. Insulin regulates the influx of glucose into the cell by upregulating the glucose transporter type 4 (GLUT4) expression on the plasma membrane. Besides, PPAR-γ also controls the metabolism of glucose in adipose tissues. Curcuma mangga Val., denoted as C. mangga, is a native Indonesian medicinal plant that has many beneficial effects, including an antidiabetic potential. PURPOSE: In this research, we aimed to disclose the hypoglycemic activity of ethanol extract of C. mangga (EECM) in 3T3-L1 fibroblasts-derived adipocyte cells in regulating glucose uptake as confirmed by the GLUT4 and PPAR-γ gene expression. METHODS: The uptake of glucose was determined using radioactive glucose, while the gene expression of GLUT4, PPAR-γ, and ß-actin was quantified using mRNA segregation and real-time quantitative reverse transcription-polymerase chain reaction (RT-qPCR). RESULTS: We discovered that EECM interventions (200 and 50 µg/mL) increased glucose uptake in lipid-laden 3T3-L1 cells by 14.75 and 8.86 fold compared to the control non-insulin, respectively (p < 0.05). At the same doses, they also increased GLUT4 mRNA expression by 8.41 and 11.18 fold compared to the control non-insulin, respectively (p < 0.05). In contrast, EECM interventions (200 and 50 µg/mL) showed lower levels of PPAR-γ mRNA expression compared to the control metformin, indicating the anti-adipogenic potentials of EECM. CONCLUSION: EECM showed hypoglycemic activity in lipid-laden 3T3-L1 cells by improving glucose ingestion into the cells, which was mediated by increased GLUT4 expression and downregulated PPAR-γ expression.

From bulk to nano-delivery of essential phytochemicals: recent progress and strategies for antibacterial resistance.

Bacterial biofilms caused by antibiotic resistance are a severe cause of infection threatening human health nowadays. The primary causes of this emerging threat are poor penetration of conventional antibiotics and the growing number of varied strains of resistant bacteria. Recently, bulk phytochemical oils have been widely explored for their potential as antibacterial agents. However, due to their poor solubility, low stability, and highly volatile properties, essential oils are not effective for in vitro and in vivo antibacterial applications and require further preparation. In this review, we discuss the recent progress and strategies to overcome the drawbacks of bulk phytochemical oils using nano-delivery, as well as the current challenges and future outlook of these nano-delivery systems against bacterial resistance.

Cosmeceutical potentials of Curcuma mangga Val. extract in human BJ fibroblasts against MMP1, MMP3, and MMP13.

Oxidative stress, the disrupted oxidation-reduction mechanism in our body, is caused by the excessive exposure of free radicals and the impaired antioxidant defenses that can accelerate skin aging. Antioxidants can be obtained from nature, which are available widely in therapeutic-rich plants, such as white saffron (Curcuma mangga Val., denoted as C. mangga). Although many pieces of evidence reveal that C. mangga contains an abundance of phenolic compounds and has antioxidative effects, its cosmeceutical potentials remain unclear. The present study aimed to disclose the unexplored antiaging potentials of C. mangga extract (CME) in oxidative stress-induced human BJ fibroblasts with a focus on collagen protection against pro-inflammatory mediators MMP1, MMP3, and MMP13. The oxidative stress-induced cells were treated with CME and curcumin at different doses. The results showed that treatment using CME (25 µg/mL) could maintain the collagen contents up to 18.45 ± 0.68 µg/mL in H2O2-treated fibroblasts (only ~26.63% reduction in collagen contents), while the figure for the negative control was the lowest (12.79 µg/mL), showing a significant reduction in collagen contents by 49.13%. In addition, the gene expression of pro-inflammatory MMPs arose significantly in BJ fibroblasts after oxidative stress induction using 200 µM H2O2, in which the expression for MMP1, MMP3, and MMP13 increased by 7.10, 38.96, and 2.69 times, respectively. Interestingly, CME treatment (100 µg/mL) could effectively inhibit MMP1, MMP3, and MMP13 gene expression by 3.65, 34.62, and 2.02 times, respectively. In conclusion, CME showed favorable antiaging activities in H2O2-treated human BJ fibroblasts as confirmed by the low levels of gene expression of MPP1, MMP3, and MMP13 after treatment with CME.

Recent advances in micro- and nano-bubbles for atherosclerosis applications.

Atherosclerosis is the most prevalent cause of cardiovascular disease-induced deaths worldwide. Micro- and nano-bubbles (MNBs) have been developed as the vehicles for detection, investigation, and drug delivery, specifically targeting atherosclerotic sites. MNBs have been clinically applied and commercialized as contrast agents because they typically respond to ultrasound for guiding and stimulating imaging. The assembly process involves some specific substrates (proteins, lipids, and polymers) to adjust their characteristics and depends upon rational designs for combined therapeutic-diagnostic (theranostic) applications. Ancillary surface modifications of MNBs enable the unification of MNBs with antibody, inflammatory markers, or genes to more specifically deliver cargos to the oxidized lipid-rich quarry area and release the payloads on demand to the lesion site. This review provides brief information on the process of fabricating MNBs and their applications in bio-nanomedicine for diagnosing and remodeling atherosclerosis.

Orally administered pressure-blanched white saffron (Curcuma mangga Val.) improves antioxidative properties and lipid profiles in vivo.

This research focused on studying the effects of orally administered pressure-blanched white saffron on the antioxidative properties and lipid profiles of wistar rats. White saffron was blanched in autoclave for 2.5, 5, 7.5, and 10 min at 100, 105, 110, 115, and 120 °C, which are equivalent to 14.71, 17.53, 20.79, 24.54, and 28.81 psia, respectively. A total of 30 male wistar rats aged four weeks were fed with a standard diet (N), oxidized peanut oil diet + unblanched white saffron (A), oxidized peanut oil diet + blanched white saffron (B), oxidized peanut oil diet + pressure-blanched white saffron (C), and oxidized peanut oil diet + aquadest (NC), for two weeks after pre-treatment with the standard diet for a week. In vivo study showed treatment with pressure-blanched white saffron could significantly improve SOD, Vitamin E, and HDL levels compared to the negative control (NC); 686.44 U/g Hb, 10.87 µg/mL, and 94.17 mg/dL versus 405.37 U/g Hb, 7.44 µg/mL, and 43.47 mg/dL, respectively. Meanwhile, treatment with pressure-blanched white saffron could significantly reduce MDA, total cholesterol, LDL, and triglyceride levels in the blood compared to the negative control (NC); 1.98 mmol/L, 108.74 mg/dL, 40.99 mg/dL, and 78.06 mg/dL versus 8.54 mmol/L, 232.46 mg/dL, 149.17 mg/dL, and 172.61 mg/dL, respectively. The results showed that pressurized blanching could significantly increase antioxidant levels of white saffron, and its dried form could improve antioxidative properties and lipid profiles in vivo.

Advanced drug-delivery systems: mechanoresponsive nanoplatforms applicable in atherosclerosis management.

Nanoplatforms have been used extensively as advanced carriers to enhance the effectiveness of drug delivery, mostly through passive aggregation provided by the enhanced permeability and retention effect. Mechanical stimuli provide a robust strategy to bolster drug delivery performance by increasing the accumulation of nanoplatforms at the lesion sites, facilitating on-demand cargo release and providing theranostic aims. In this review, we focus on recent advances of mechanoresponsive nanoplatforms that can accomplish targeted drug delivery, and subsequent drug release, under specific stimuli, either endogenous (shear stress) or exogenous (magnetic field and ultrasound), to synergistically combat atherosclerosis at the molecular level.

Transforming stealthy to sticky nanocarriers: a potential application for tumor therapy.

Nanomedicine has shown remarkable progress in preclinical studies of tumor treatment. Over the past decade, scientists have developed various nanocarriers (NCs) for delivering drugs into the tumor area. However, the average amount of accumulated drugs in tumor sites is far from satisfactory. This limitation is strongly related to the corona formation during blood circulation. To overcome this issue, NCs should be designed to become highly stealthy by modifying their surface charge. However, at the same time, stealthy effects not only prevent protein formation but also alleviate the cellular uptake of NCs. Therefore, it is necessary to develop NCs with switchable properties, which are stealthy in the circulation system and sticky when arriving at tumor sites. In this review, we discuss the recent strategies to develop passive and active charge-switchable NCs, known as chameleon-like drug delivery systems, which can reversibly transform their surface from stealthy to sticky and have various designs.

Atherosclerosis Treatment with Stimuli-Responsive Nanoagents: Recent Advances and Future Perspectives.

Atherosclerosis is the root of approximately one-third of global mortalities. Nanotechnology exhibits splendid prospects to combat atherosclerosis at the molecular level by engineering smart nanoagents with versatile functionalizations. Significant advances in nanoengineering enable nanoagents to autonomously navigate in the bloodstream, escape from biological barriers, and assemble with their nanocohort at the targeted lesion. The assembly of nanoagents with endogenous and exogenous stimuli breaks down their shells, facilitates intracellular delivery, releases their cargo to kill the corrupt cells, and gives imaging reports. All these improvements pave the way toward personalized medicine for atherosclerosis. This review systematically summarizes the recent advances in stimuli-responsive nanoagents for atherosclerosis management and its progress in clinical trials.

Targeted polyethylenimine/(p53 plasmid) nanocomplexes for potential antitumor applications.

The development of the tumor-targeting ability of nanocarriers is of paramount importance for gene delivery into tumor lesions as well as to avoid biotoxicity. Here we report the synthesis of the polyethyleneimine-fluorescein isothiocyanate-folic acid (PEI-FITC-FA) polymer, which could condense the tumor suppressor pp53 to form nanocomplexes. These targeted nanocomplexes exhibited favorable physical properties including a small size of <100 nm, exploiting the enhanced permeability and retention effect and tumor-targeting ability by binding to the overexpressed FA receptors on tumor cell surfaces. In addition, once the nanocomplexes are accumulating in the tumor tissue, the target functional ligand, FA, can selectively recognize the over-expressed FA receptor and subsequently remain on the tumor cell surface, which can significantly promote the tumor cell uptake because of the time- and concentration-dependent internalization caused by the enhanced interaction between nanocomplex and tumor cell. Our results indicated that PEI-FITC-FA/pp53 nanocomplexes could be efficiently delivered into tumor cells, and subsequently induce tumor cell apoptosis. Thus, the targeted cationic polymer PEI-FITC-FA could be used as an advanced nanocarrier for gene delivery.