Multifunctional CaCO3@Cur@QTX125@HA nanoparticles for effectively inhibiting growth of colorectal cancer cells.

Colorectal cancer (CRC) is a major cause of cancer-related deaths in humans, and effective treatments are still needed in clinical practice. Despite significant developments in anticancer drugs and inhibitors, their poor stability, water solubility, and cellular membrane permeability limit their therapeutic efficacy. To address these issues, multifunctional CaCO3 nanoparticles loaded with Curcumin (Cur) and protein deacetylase (HDAC) inhibitor QTX125, and coated with hyaluronic acid (HA) (CaCO3@Cur@QTX125@HA), were prepared through a one-step gas diffusion strategy. Dynamic light scattering (DLS), transmission electron microscopy (TEM), and scanning electron microscopy (SEM) showed that CaCO3@Cur@QTX125@HA nanoparticles have uniform spherical morphology and elemental distribution, with diameters around 450 nm and a Zeta potential of - 8.11 mV. The controlled release of Cur from the nanoparticles was observed over time periods of 48 h. Cellular uptake showed that CaCO3@Cur@QTX125@HA nanoparticles were efficiently taken up by cancer cells and significantly inhibited their growth. Importantly, CaCO3@Cur@QTX125@HA nanoparticles showed specific inhibitory effects on CRC cell growth. Encouragingly, CaCO3@Cur@QTX125@HA nanoparticles successfully internalized into CRC patient-derived organoid (PDO) models and induced apoptosis of tumor cells. The multifunctional CaCO3@Cur@QTX125@HA nanoparticles hold promise for the treatment of CRC.

Tumor-targeted paclitaxel-loaded folate conjugated poly(ethylene glycol)-poly(L-lactide) microparticles produced by supercritical fluid technology.

The new biodegradable diblock copolymers poly(ethylene glycol)-poly(L-lactide) (PEG-PLLA) were synthesized and were chemically conjugated with folate (FA) in the PEG terminal ends to form FA-PEG-PLLA. Then the hydrophobic drug paclitaxel (PTX) loaded microparticles (PTX/FA-PEG-PLLA) were produced via solution enhanced dispersion by supercritical fluids (SEDS). These microparticles exhibited sphere-like shape by scanning electron microscopy observation and showed narrow hydrodynamic size distributions by dynamic light scattering measurement. Drug loading of PTX loaded microparticles was about 7-9% and the encapsulation efficiency of PTX loaded microparticles was about 18-23%. Flow cytometry and confocal laser scanning microscope analyses revealed that fluorescein isothiocyanate labeled FA conjugated microparticles presented significantly higher cellular uptake than FA-free group due to the FA-receptor-mediated endocytosis. In vitro cytotoxicity evaluation indicated that FA-PEG-PLLA expressed negligible cytotoxicity to mouse fibroblasts L929 cells. Moreover, PTX/FA-PEG-PLLA microparticles exhibited much higher anti-cancer efficacy than PTX/PEG-PLLA microparticles against human ovarian cancer SKOV3 cells. Nude mice xenografted with SKOV3 cells were used in biodistribution studies, the results indicated that an increased amount of PTX was accumulated in the tumor tissue deal with PTX/FA-PEG-PLLA microparticles. These results collectively suggested that PTX/FA-PEG-PLLA microparticles prepared by SEDS would have potential in anti-tumor applications as a tumor-targeted drug delivery formulation.

Tumor microenvironment targeting for glioblastoma multiforme treatment via hybrid cell membrane coating supramolecular micelles.

Glioblastoma multiforme (GBM) is one of the most common primary intracranial tumors in the central nervous system with poor prognosis, high invasiveness, risk of recurrence and low survival rate. Thus, it is urgent and vital to develop drug effective delivery systems that efficiently to traverse the blood-brain barrier and targeted transport therapeutic agents into the GBM tumor site for the treatment of brain tumors. Recently, amphiphilic cucurbit[7]uril-polyethylene glycol-hydrophobic Chlorin e6 (CB[7]-PEG-Ce6) polymer was designed, prepared, and self-assembled into micells (CPC) in an aqueous solution, and chemo drug methyl-triazeno-imidazole-carboxamide (MTIC), loaded into the cavity of CB[7] was subsequently coated with hybrid membrane mUMH (HMC3 membrane: macrophage membrane: U87MG membrane = 1:1:2) to afford mUMH@CPC@MTIC. The surface hybrid membrane mUMH potentially enhance the targeted delivery of CPC@MTIC to GBM tissue. Bioactive MTIC was released from the cavity of CB[7] in response to the high spermine level in GBM tumor microenvironments for effective tumor chemotherapy. The biomimetic mUMH@CPC@MTIC exhibited superior antitumor efficacy against GBM in mice. These findings provide new strategies for the design of biomimetic nanoparticle-based drug delivery systems and promising therapy of GBM.

Macrophage Membrane-Encapsulated Dopamine-Modified Poly Cyclodextrin Multifunctional Biomimetic Nanoparticles for Atherosclerosis Therapy.

Atherosclerotic plaques exhibit high cholesterol deposition and oxidative stress resulting from high reactive oxygen species (ROS). These are the major components in plaques and the main pro-inflammatory factor. Therefore, it is crucial to develop an effective therapeutic strategy that can simultaneously address the multiple pro-inflammatory factors via removing cholesterol and inhibiting the overaccumulated ROS. In this study, we constructed macrophage membrane-encapsulated biomimetic nanoparticles (MM@DA-pCD@MTX), which not only alleviate cholesterol deposition at the plaque lesion via reverse cholesterol transport but also scavenge the overaccumulated ROS. ß-Cyclodextrin (ß-CD) and the loaded methotrexate (MTX) act synergistically to induce cholesterol efflux for inhibiting the formation of foam cells. Among them, MTX up-regulated the expression of ABCA1, CYP27A1, and SR-B1. ß-CD increased the solubility of cholesterol crystals. In addition, the ROS scavenging property of dopamine (DA) was perfectly preserved in MM@DA-pCD@MTX, which could scavenge the overaccumulated ROS to alleviate the oxidative stress at the plaque lesion. Last but not least, MM-functionalized "homing" targeting of atherosclerotic plaques not only enables the targeted drug delivery but also prolongs in vivo circulation time and drug half-life. In summary, MM@DA-pCD@MTX emerges as a potent, multifunctional therapeutic platform for AS treatment, offering a high degree of biosafety and efficacy in addressing the complex pathophysiology of atherosclerosis.

Engineered Cell Membrane-Coated Nanoparticles: New Strategies in Glioma Targeted Therapy and Immune Modulation.

Gliomas, the most prevalent primary brain tumors, pose considerable challenges due to their heterogeneity, intricate tumor microenvironment (TME), and blood-brain barrier (BBB), which restrict the effectiveness of traditional treatments like surgery and chemotherapy. This review provides an overview of engineered cell membrane technologies in glioma therapy, with a specific emphasis on targeted drug delivery and modulation of the immune microenvironment. This study investigates the progress in engineered cell membranes, encompassing physical, chemical, and genetic alterations, to improve drug delivery across the BBB and effectively target gliomas. The examination focuses on the interaction of engineered cell membrane-coated nanoparticles (ECM-NPs) with the TME in gliomas, emphasizing their potential to modulate glioma cell behavior and TME to enhance therapeutic efficacy. The review further explores the involvement of ECM-NPs in immunomodulation techniques, highlighting their impact on immune reactions. While facing obstacles related to membrane stability and manufacturing scalability, the review outlines forthcoming research directions focused on enhancing membrane performance. This review underscores the promise of ECM-NPs in surpassing conventional therapeutic constraints, proposing novel approaches for efficacious glioma treatment.

[Determination of chlorpropham residues in animal-derived foods by solid phase extraction and ultra-high performance liquid chromatography-tandem mass spectrometry].

Chlorpropham is a plant growth regulator and a herbicide. It is commonly used in the post-harvest treatment of potato to inhibit germination. It can also be used for flower thinning and fruit thinning of fruit trees, and for controlling annual gramineous weeds and a few broad-leaved weeds. Improper or excessive use of chlorpropham in crop cultivation will affect the safety of animal-derived food and impair human health through the food chain and water cycle. Therefore, accurate quantification of chlorpropham is imperative for risk assessment and mitigating risks to food safety. A method based on solid phase extraction and ultra-high performance liquid chromatography-tandem mass spectrometry (SPE-UHPLC-MS/MS) was established for the determination of chlorpropham in animal-derived food. First, the pretreatment conditions were optimized. To purify the samples and remove impurities, SPE column cartridges with different packing materials such as PXC, PXA, Florisil, and PLS were investigated. Based on the retention of chlorpropham, the ProElut PLS SPE column was selected as the pretreatment purification column. The washing solution and eluents were then optimized. When water was used as the washing solution, chlorpropham remained adsorbed on the SPE column and was not eluted along with other water-soluble substances. When the proportion of acetonitrile exceeded 40%, chlorpropham adsorbed on the filler of the SPE column could be gradually washed down. Acetonitrile-water solution(30∶70, v/v) was used for washing the SPE column. The elution ability of seven eluents for chlorpropham on the SPE column was then investigated. Among them, pure methanol, pure acetonitrile, and 1% (v/v) formic acid-methanol showed better elution effect. Considering that acetonitrile was used in the sample extraction, it was chosen as the mobile phase eluent. Subsequently, the chromatographic conditions and MS parameters were optimized. By examining the ionization cracking of chlorpropham, the quasimolecular ions and corresponding fragmentations in the chlorpropham primary MS were determined. The separation effect of three C18 columns was investigated. Based on the retention ability and peak effect of chlorpropham on the column, the Agilent ZORBAX SB-C18 (150 mm×2.1 mm, 5 µm) column was used for chlorpropham separation. The response of chlorpropham in the positive and negative ionization modes was investigated and optimized. The results showed that the response was better in the positive ion mode than that in the negative ion mode. After optimizing the chromatographic conditions and MS parameters, the sensitivity of the method was improved. Finally, the analytes were separated on the Agilent ZORBAX SB-C18 (150 mm×2.1 mm, 5 µm) under a gradient elution program using acetonitrile and 0.2% (v/v) formic acid aqueous solution as the mobile phases. The analytes were detected in the multiple reaction monitoring (MRM) mode under positive electrospray ionization (ESI+) conditions. The standard curve solutions were prepared using the matrix solution and quantified by the external standard method. The results showed a good linear relationship in the range of 0.5-100.0 µg/L, with correlation coefficients (r2) greater than 0.9929. The limit of quantification (LOQ) of this method was 3 µg/kg (S/N ≥ 10). At three spiked levels (0.003, 0.006, and 0.060 mg/kg) in 13 animal-derived foods (pork, milk, beef, chicken, duck, egg, chicken gizzard, duck egg, pork kidney, pork liver, beef liver, mutton, duck gizzard), the average recoveries were in the range of 74.9% to 97.6%, and the RSDs were in the range of 2.9% to 9.5% (n=6). Sixty batches of animal-derived food available on the market were analyzed by the developed method, and chlorpropham was not detected in any of these foods. The developed method is rapid, sensitive, and accurate, and it is suitable for the qualitative and quantitative detection of chlorpropham in a variety of animal-derived foods.

Genetic Engineering of Adipose-Derived Stem Cells Using Biodegradable and Lipid-Like Highly Branched Poly(ß-amino ester)s.

Biodegradable and lipid-like highly branched poly(ß-amino ester)s, HPAESA, were developed to enhance the biological functions of adipose-derived stem cells by gene transfection. Biodegradability reduces the cytotoxicity of HPAESA and enables controlled DNA release. Lipid mimicry enhances cellular uptake and endosomal escape of HPAESA/DNA polyplexes. HPAESA are able to transfect rat adipose-derived stem cells (rADSs) and human ADSCs (hADSCs) with orders of magnitude higher efficiency than commercial gene transfection reagents, with cell viability exceeding 90%. Most importantly, HPAESA can effectively transfer the nerve growth factor (NGF)-encoding plasmid to rADSCs and induce high NGF secretion, which significantly promotes neurite outgrowth of PC12 cells.

Targeted delivery and enhanced uptake of chemo-photodynamic nanomedicine for melanoma treatment.

Nanoparticles (NPs) modified with targeting ligands have often shown great potential in targeted drug delivery for tumor therapy. However, the clearance of NPs by the monocyte-phagocyte system (MPS) and the relatively low cellular uptake by tumor cells have significantly limited the antitumor efficacy of a variety of nanomedicines. Tumor microenvironment-mediated multidrug resistance also reduces the antitumor efficacy of internalized nanomedicines. Herein, we developed an innovative nanomedicine for combined chemo-photodynamic therapy of melanoma through targeted drug delivery and significantly improved the cellular uptake of the nanomedicine through the charge-reversal phenomenon. An amphiphilic platinum (IV)-polyethylenimine-chlorin e6 (Pt(IV)-PEI-Ce6) polymer was designed, prepared, and self-assembled into NPs (PPC) in an aqueous solution, and these NPs were subsequently coated with hyaluronic acid (HA) to afford PPC@HA. The surface-coated HA provided PPC with a negatively charged surface potential to reduce the clearance by the MPS during systemic circulation and enhanced the targeted delivery of PPC to CD44-overexpressing melanoma cells. Upon accumulation in the tumor site, hyaluronidase overexpressed in the tumor induced HA degradation to release the positively charged PPC, resulting in an increased internalization of PPC into tumor cells. Bioactive Pt(II) was released in response to high glutathione level in the tumor cells for effective tumor chemotherapy. Under 650 nm laser irradiation, Ce6 produced reactive oxygen species (ROS), thus driving photodynamic therapy. Finally, PPC@HA exhibited combined photodynamic-chemotherapeutic antitumor efficacy against the melanoma cells in mice. STATEMENT OF SIGNIFICANCE: Tumors are one of the greatest threats to human health, and chemotherapy has been one of the most common therapeutic modalities for treating tumors; however, many challenges related to chemotherapy remain, such as low delivery efficiency, side effects, and unsatisfactory therapeutic efficacy. Nanomedicines modified with targeting ligands have often shown great potential in improving targeted drug delivery for tumor therapy; however, the clearance of nanomaterials by the monocyte-phagocyte system and the relatively low cellular uptake by tumor cells have significantly limited the antitumor efficacy of a variety of nanomedicines. Herein, we developed a novel charge-reversal-based, hyaluronic acid-coated, Pt(IV) prodrug and chlorin e6-based nanomedicine to improve systemic circulation and targeted accumulation of the nanomedicine in the tumor tissue and to enhance its intracellular uptake. This nanomedicine may provide a potential new platform to improve the drug content inside tumor cells and to effectively inhibit tumor growth through combined chemotherapy and photodynamic therapy.

Cyclic poly(ß-amino ester)s with enhanced gene transfection activity synthesized through intra-molecular cyclization.

Topological structure plays a critical role in gene delivery of cationic polymers. Cyclic poly(ß-amino ester)s (CPAEs) are successfully synthesized via sequential Michael addition and free radical initiating ring-closure reaction. The CPAEs exhibit superior gene transfection efficiency and safety profile compared to their linear counterparts.

Supramolecular micelles as multifunctional theranostic agents for synergistic photodynamic therapy and hypoxia-activated chemotherapy.

Photodynamic therapy (PDT), where a photosensitizer (under light irradiation) converts molecular oxygen to singlet oxygen to elicit programmed cell death, is a promising cancer treatment modality with a high temporal and spatial resolution. However, only limited cancer treatment efficacy has been achieved in clinical PDT due to the hypoxic conditions of solid tumor microenvironment that limits the generation of singlet oxygen, and PDT process often leads to even more hypoxic microenvironment due to the consumption of oxygens during therapy. Herein, we designed novel supramolecular micelles to co-deliver photosensitizer and hypoxia-responsive prodrug to improve the overall therapeutic efficacy. The supramolecular micelles (CPC) were derived from a polyethylene glycol (PEG) system dually tagged with hydrophilic cucurbit[7]uril (CB[7]) and hydrophobic Chlorin e6 (Ce6), respectively on each end, for synergistic antitumor therapy via PDT of Ce6 and chemotherapy of a hypoxia-responsive prodrug, banoxantrone (AQ4N), loaded into the cavity of CB[7]. In addition, CPC was further modularly functionalized by folate (FA) via strong host-guest interaction between folate-amantadine (FA-ADA) and CB[7] to produce a novel nanoplatform, AQ4N@CPC-FA, for targeted delivery. AQ4N@CPC-FA exhibited enhanced cellular uptake, negligible cytotoxicity and good biocompatibility, and improved intracellular reactive oxygen species (ROS) generation efficiency. More importantly, in vivo evaluation of AQ4N@CPC-FA revealed a synergistic antitumor efficacy between PDT of Ce6 and hypoxia-activated chemotherapy of AQ4N (that can be converted to chemotherapeutic AQ4 for tumor chemotherapy in response to the strengthened hypoxic tumor microenvironment during PDT treatment). This study not only provides a new nanoplatform for synergistic photodynamic-chemotherapeutic treatment, but also offers important new insights to design and development of multifunctional supramolecular drug delivery system. STATEMENT OF SIGNIFICANCE: Photodynamic therapy (PDT) has exhibited a variety of advantages for cancer phototherapy as compared to traditional chemotherapy. However, the unsatisfactory therapeutic efficacy by PDT alone as a result of the enhanced tumor hypoxia during PDT has limited its clinical application. Herein, we designed multifunctional supramolecular micelles to co-deliver photosensitizer and hypoxia-responsive prodrug to improve the overall therapeutic efficacy. The supramolecular micelles are biocompatible and possess strong red absorption, controlled drug release profile, and ultimately enhanced therapeutic outcome via PDT-chemotherapy. This study not only provides a new nanoplatform for synergistic photodynamic-chemotherapeutic treatment of cancer, but also offers important new insights to design and development of multifunctional supramolecular drug delivery tool for multi-modality cancer therapy.

[Determination of a new sweetener advantame in food by ultra-high performance liquid chromatography-tandem mass spectrometry].

A new ultra-high performance liquid chromatography-tandem mass spectrometry with triple quadrupole method was developed, and validated for screening and confirmation of the new sweetener advantame in food. The sample was extracted with methanol-water (50:50, v/v), followed by centrifugation. The supernatant was then filtered through a membrane filter. The determinant was performed on Agilent SB-C18 (150 mm×2.1 mm, 5 µm) column with gradient elution. The target compound was carried out by positive electrospray ionization (ESI+) under the multiple reaction monitoring (MRM) mode. While the qualitative screening was achieved with retention time and qualitative ion, the confirmation analysis was achieved with peak area and quantitative ion. The results showed that the limit of detection (LOD, S/N ≥ 3) and the limit of quantification (LOQ, S/N ≥ 10)were 0.03 and 0.10 mg/kg, respectively. The average spiked recoveries showed a variation from 80.3 to 98.0% for the three fortification levels. The results showed good linear relationships in the range of 0.01-1.0 mg/L of advantame with correlation coefficients above 0.997. This procedure was noticeably rapid, simple, sensitive, and accurate. Therefore, this method is suitable for batch determination of the new sweetener advantame in beverages, yogurt, and jelly.

Thermosensitive star polymer pompons with a core-arm structure as thermo-responsive controlled release drug carriers.

In contrast with traditional chemotherapy, controlled drug delivery systems provide many advantages. Herein, a thermosensitive star polymer pompon with a core-arm structure was synthesized using a grafting-on method as a thermo-responsive controlled release drug carrier. Single-chain cyclized/knotted poly tetra(ethylene glycol) diacrylate (polyTEGDA) was used as the hydrophobic core, and thermosensitive linear poly(N-isopropylacrylamide-co-N-methylolacrylamide) (poly(NIPAM-co-NMA)) was selected as the hydrophilic arm. Below or above its lower critical solution temperature (LCST), the linear poly(NIPAM-co-NMA) grafted onto the polyTEGDA core adopted a stretched or curled status, respectively, then the drug could be loaded in or extruded out. The LCST of star polyTEGDA-b-poly(NIPAM-co-NMA) was adjusted to slightly above body temperature (37 °C). The antitumor drug doxorubicin (DOX) was successfully loaded into the pompons with a high loading capacity of 19.45%. The cumulative release of DOX from loaded pompons in vitro for 72 hours was 71% and 20.7% at 42 °C and 37 °C, respectively, indicating that the excellent temperature-controlled release characteristics result from the unique thermo-responsive extrusion effect. Moreover, DOX loaded polyTEGDA-b-poly(NIPAM-co-NMA) pompons achieved better antitumor ability against ovarian carcinoma SKOV3 cells at 42 °C compared with that at 37 °C. These results suggest that star polyTEGDA-b-poly(NIPAM-co-NMA) pompons have considerable promise as thermo-responsive controlled drug delivery carriers.

Thermo- and pH-Responsive, Coacervate-Forming Hyperbranched Poly(ß-amino ester)s for Selective Cell Binding.

We report a new type of thermo- and pH-responsive, coacervate-forming highly degradable polymer-hyperbranched poly(ß-amino esters) (HPAEs) and its selective cell binding behaviors. The HPAEs were synthesized from 5-amino-1-pentanol (S5) and trimethylolpropane ethoxylate triacrylate (TMPETA) via an A2+B3 type Michael addition. The existence of multiple hydrogen bond pairs as well as tertiary amines makes the S5-TMPETA polymers manifest temperature- and pH-dependent phase transition. By varying the length of the ethylene glycol (EG) spacers in the TMPETA, polymer molecular weight, concentration, and pH value, the phase transition of the S5-TMPETA can be easily tuned in aqueous and buffer solutions, as evidenced by UV-vis spectroscopy and DLS measurements. Especially, the S5-TMPETA prepared from S5 and trimethylolpropane ethoxylate triacrylate 692 (S5-TMPETA692) shows a lower critical solution temperature (LCST) around 33 °C, above which the S5-TMPTEA can form coacervate particles able to encapsulate functional molecules effectively. Importantly, when incubation with HeLa cells, the S5-TMPTETA692 exhibits a temperature- and pH-responsive selective cell binding behaviors. In addition, the S5-TMPETA are highly hydrolyzable and elicit negligible cytotoxicity. This new type of "smart" polymer should find use in a variety of biomedical applications.

Star Poly(ß-amino esters) Obtained from the Combination of Linear Poly(ß-amino esters) and Polyethylenimine.

Composed of a three-dimensional structure with a central core and multiple radiating linear "arms", star polymers represent a significant type of branched macromolecular architectures. Due to the spatially defined core-shell-periphery architecture, star polymers have demonstrated their superiority in a variety of biomedical applications such as drug/gene delivery, molecular imaging, antibacterial agents, and so on. In this paper, we report the successful synthesis of a new type of star-shape poly(ß-amino esters) with low molecular weight PEI as core and linear PAE (LPAE) as arms. This new star-PAE exhibits low cytotoxicity and high gene transfection efficacy. Star-PAE achieved between 264-fold and 14781-fold higher gene transfection efficiency of primary rat adipose derived mesenchymal stem cells in comparison with studies performed with the individual PEI and LPAE, respectively. The results suggest that star-PAE is a promising nonviral gene delivery vector.

Development of Branched Poly(5-Amino-1-pentanol-co-1,4-butanediol Diacrylate) with High Gene Transfection Potency Across Diverse Cell Types.

One of the most significant challenges in the development of polymer materials for gene delivery is to understand how topological structure influences their transfection properties. Poly(5-amino-1-pentanol-co-1,4-butanediol diacrylate) (C32) has proven to be the top-performing gene delivery vector developed to date. Here, we report the development of branched poly(5-amino-1-pentanol-co-1,4-butanediol diacrylate) (HC32) as a novel gene vector and elucidate how the topological structure affects gene delivery properties. We found that the branched structure has a big impact on gene transfection efficiency resulting in a superior transfection efficiency of HC32 in comparison to C32 with a linear structure. Mechanistic investigations illustrated that the branched structure enhanced DNA binding, leading to the formation of toroidal polyplexes with smaller size and higher cationic charge. Importantly, the branched structure offers HC32 a larger chemical space for terminal functionalization (e.g., guanidinylation) to further enhance the transfection. Moreover, the optimized HC32 is capable of transfecting a diverse range of cell types including cells that are known to be difficult to transfect such as stem cells and astrocytes with high efficiency. Our study provides a new insight into the rational design of poly(ß-amino ester) (PAE) based polymers for gene delivery.