Multifunctional Covalent Organic Framework-Based Microneedle Patch for Melanoma Treatment.

Melanoma is resistant to conventional chemotherapy and radiotherapy. Therefore, it is essential to develop a targeted, low-toxic, and minimally invasive treatment. Here, DTIC/ICG-Fe3O4@TpBD BSP/HA microneedles (MNs) were designed and fabricated, which can enhance targeting to melanoma and perform photothermal therapy (PTT) and chemotherapy simultaneously to synergistically exert anticancer effects. The system consisted of magnetic nanoparticles (DTIC/ICG-Fe3O4@TpBD), dissoluble matrix (Bletilla polysaccharide (BSP)/hyaluronic acid (HA)), and a polyvinyl alcohol backing layer. Due to the good magnetic responsiveness of Fe3O4@TpBD, dacarbazine (DTIC) and indocyanine green (ICG) can be better targeted to the tumor tissue and improve the therapeutic effect. BSP and HA have good biocompatibility and transdermal ability, so that the MNs can completely penetrate the tumor tissue, be dissolved by the interstitial fluid, and release DTIC and ICG. Under near-infrared (NIR) light irradiation, ICG converts light energy into thermal energy and induces ablation of B16-OVA melanoma cells. In vivo results showed that DTIC/ICG-Fe3O4@TpBD BSP/HA MNs combined with chemotherapy and PTT could effectively inhibit the growth of melanoma without tumor recurrence or significant weight loss in mice. Therefore, DTIC/ICG-Fe3O4@TpBD BSP/HA MNs are expected to provide new ideas and therapeutic approaches for the clinical treatment of melanoma.

Improved pharmacodynamics of epidermal growth factor via microneedles-based self-powered transcutaneous electrical stimulation.

Epidermal growth factor is an excellent drug for promoting wound healing; however, its conventional administration strategies are associated with pharmacodynamic challenges, such as low transdermal permeability, reduction, and receptor desensitization. Here, we develop a microneedle-based self-powered transcutaneous electrical stimulation system (mn-STESS) by integrating a sliding free-standing triboelectric nanogenerator with a microneedle patch to achieve improved epidermal growth factor pharmacodynamics. We show that the mn-STESS facilitates drug penetration and utilization by using microneedles to pierce the stratum corneum. More importantly, we find that it converts the mechanical energy of finger sliding into electricity and mediates transcutaneous electrical stimulation through microneedles. We demonstrate that the electrical stimulation applied by mn-STESS acts as an "adjuvant" that suppresses the reduction of epidermal growth factor by glutathione and upregulates its receptor expression in keratinocyte cells, successfully compensating for receptor desensitization. Collectively, this work highlights the promise of self-powered electrical adjuvants in improving drug pharmacodynamics, creating combinatorial therapeutic strategies for traditional drugs.

UHPLC-ESI-MS/MS Quantitative Analyses of Multicomponent Hu Gan Tablets.

Nowadays, the analysis of the multi-components in Chinese patent medicine prescriptions is being paid more attention. Therefore, in this study for the first time, a simple, rapid ultrahigh performance liquid chromatography-electrospray ionization tandem mass spectrometry (UHPLC-ESI-MS/MS) method was established for simultaneous determination of 18 active compounds in a Chinese patent medicine of Hu Gan tablets (HGT) from different pharmaceutical factories in China. This task has met great emerging challenges from not only structural complexities and similarities but also co-occurrence of water-soluble and fat-soluble components in HGT. UPLC-ESI-MS/MS was put forward to solve the problems. It was operated in both positive and negative mode using multiple reaction monitoring (MRM). The mobile phase was 0.1% formic acid in water (A) -0.1% formic acid in acetonitrile (B) with linear gradient elution at a flow rate of 0.2 mL/min, run for a total of 12.0 min. The optimized method used provided short analysis time and good linearity (R2 > 0.99), and intra- and inter-day precision (relative standard deviation (RSD) < 4.00%) with good accuracy (94.89-110.03%) and recovery (70.00-126.09%). The results indicate the method could be practically used for quality guarantee of HGT and might also be useful for further studies.

Rapid screening and characterization of triterpene saponins in Acanthopanax senticosus leaves via untargeted MSAll and SWATH techniques on a quadrupole time of flight mass spectrometry.

This paper focused on untargeted MSAll, also called MSE, and sequential window acquisition of all theoretical fragment-ion spectra (SWATH) fragmentations for comprehensive structural characterization of triterpene saponins (TSs) in leaves of Acanthopanax senticosus through ultra-high performance liquid chromatography electrospray quadrupole time-of-flight mass spectrometry (UPLC-ESI-QTOF). [M+H]+, [M + NH4]+ and [M + Na]+ precursor ions and the corresponding fragment ions were collected simultaneously in energy-resolved MSAll. SWATH fragmentation was applied as a comparable and complementary method for resolving co-eluting species. A workflow based on MSAll and SWATH fragmentations was constructed for comprehensive structural characterization and rapid discovery of TSs in leaves of A. senticosus. As a result, 89 TSs, along with 14 sapogenins, were unambiguously characterized or tentatively identified. Of these, 33 compounds were characterized as potentially new compounds, including the first report of malonyl-saponin in genus Acanthopanax. This study aimed to systematically analyze TSs in leaves of A. senticosus, and the results are significant for the utilization of A. senticosus leaves.

Self-implanted tiny needles as alternative to traditional parenteral administrations for controlled transdermal drug delivery.

Controlled drug-delivery systems have potential as substitutes for traditional medication systems due to the advantages in safety, efficacy, and patient compliance that these long-acting dosage forms provide. In this context, the present study focus on the development of self-implanted hyaluronic acid (HA) tiny needles that encapsulate ivermectin (IVM)-poly (lactic-co-glycolic acid) (PLGA) microparticles for controlled transdermal IVM release to treat parasitic diseases. The fabricated tiny needles involved matching portable applicator have potentially able for self-administration by patients without intense pain or complexity of current controlled-release devices. The biodegradable IVM-loaded PLGA microparticles were prepared and encapsulated within the tip of dissolving HA tiny needles to achieve high delivery efficiency. The drug loading of tiny needles might be controlled by varying the repeat time of filling or pressing processes. In-vitro tests showed that the tiny needles have sufficient mechanical strength to be inserted into skin within seconds and, next rapidly dissolved to release the loaded drug carriers into subcutaneous tissues for intradermal sustained IVM release. With the in-vivo test in rats, the insertion site recovered barrier property within 3 h. In comparison to traditional hypodermic injection or implantation of controlled-release systems, the proposed polymer tiny needles can be considered as a promising device for controlled transdermal drug delivery.

A generic strategy based on gas phase decomposition of protonated and ammoninted precursors producing predictable MRM-MS ion pairs and collision energies for direct analysis of plant triterpene glycosides.

Optimization of multiple reaction monitoring mass spectrometry (MRM-MS) parameters of triterpene glycosides (TGs) using traditional infusion methods remains to be labor-intensive. However, it was found that mild gas phase decompositions of protonated and ammoninted precursors (DPAP) of TGs could produce a series of abundant dehydrated product ions of aglycones ([A+H-nH2O]+ (n = 0, 1, 2, 3…)) with high efficiency and stability. Based on these considerations and findings, an innovative ESI+-MRM-DPAP-MS strategy was devised on a QTRAP 4000 instrument allowing for rapid the qualitative and quantitative analysis of plant TGs. A detailed study of 85 model compounds from 20 herbal medicines was implemented for validation and evaluation of the ESI+-MRM-DPAP-MS strategy proposed. The central composition design confirmed that collision energy (CE) played more significant roles than declustering potentials (DP) for the formation of these Q1/Q3 ion pairs based on MRM-DPAP-MS. It is also noted that Q1 and Mw were the most important factors for the prediction of CE values by a partial least square regression model. Here, we demonstrated this generic workflow and its merits in: (1) early prediction and selection of MRM ion pairs, no matter which type of TGs, employing a new-found Q1/Q3 calculation formula (Q1=[M+H/NH4]+ and Q3= [A+H-nH2O]+ (n = 0, 1, 2, 3…)); (2) direct determination of practicable CE values using TGs-specific CE-estimating linear equations; (3) appearances of excellent sensitivity, stability and repeatability through real application in Aralia elata, Panax notoginseng and Caulophyllum robustum; (4) seamless application of optimal CE parameters in other triple quadrupole MS instruments such as Thermo TSQ Quantum Ultra. The ESI+-MRM- DPAP-MS may service as an effective and feasible approach for analytical characterization of biological TGs from herbal medicines.

Quality Analysis of American Ginseng Cultivated in Heilongjiang Using UPLC-ESI--MRM-MS with Chemometric Methods.

American ginseng (Panax quinquefolium) has long been cultivated in China for the function food and medicine. Here, ultra-high performance liquid chromatography was coupled with electrospray ionization and triple quadrupole mass spectrometry (UPLC-ESI--TQ-MS) for simultaneous detection of 22 ginsenosides in American ginseng cultivated in Mudanjiang district of Heilongjiang. The extraction conditions also were optimized by a Box Behnken design experiment. The optimized result was 31.8 mL/g as ratio of liquid to raw materials, 20.3 min of extraction time, and 235.0 W of extraction powers. The quantitative MS parameters for these 22 compounds were rapidly optimized by single factor experiments employing UPLC-ESI--multiple reaction monitoring or multiple ion monitoring (MRM/MIM) scans. Furthermore, the established UPLC-ESI--MRM-MS method showed good linear relationships (R² > 0.99), repeatability (RSD < 3.86%), precision (RSD < 2.74%), and recovery (94⁻104%). This method determined 22 bioactive ginsenosides in different parts of the plant (main roots, hairy roots, rhizomes, leaves, and stems) and growth years (one year to four years) of P. quinquefolium. The highest total content of the 22 analytes was in the hairy roots (1.3 × 105 µg/g) followed by rhizomes (7.1 × 104 µg/g), main roots (6.5 × 104 µg/g), leaves (4.2 × 104 µg/g), and stems (2.4 × 104 µg/g). Finally, chemometric methods, hierarchical clustering analysis (HCA) and partial least squares discrimination analysis (PLS-DA), were successfully used to classify and differentiate American ginseng attributed to different growth years. The proposed UPLC-ESI--MRM-MS coupled with HCA and PLS-DA methods was elucidated to be a simple and reliable method for quality evaluation of American ginseng.

In vitro and in vivo assessment of polymer microneedles for controlled transdermal drug delivery.

Microneedles (MNs) system for transdermal drug delivery has the potential to improve therapeutic efficacy, proving an approach that is more convenient and acceptable than traditional medication systems. This study systematically researched dissolving polymer MNs fabricated from various common FDA-approved biocompatible materials, including gelatine, chitosan, hyaluronic acid (HA) and polyvinyl alcohol (PVA). Upon application of MN patches to the porcine cadaver skin, the MNs effectively perforated the skin and delivered drugs to subcutaneous tissue on contact with the interstitial fluid. Both the in vitro and in vivo drug release tests showed the similar trends but different release rates among the prepared MNs. Interestingly, the drug-release kinetics of PVA MNs were able to be altered by changing the molecular weight. To evaluate the feasibility using the proposed MNs for treating diabetes, an in vivo insulin absorption study in diabetic mice was performed. The results showed different insulin release properties of MNs fabricated from various kinds of polymer, leading to different decrease in blood glucose levels. We made a systematic and comprehensive study of some drug-loaded polymer MNs, and anticipated that dissolving polymer MNs have potential to improve therapeutic efficacy through controlled drug release.

UPLC-QTOF-MSE -based diagnostic product ion filtering to unveil unstable C6 -C2 glucoside conjugates in Forsythia suspensa.

Forsythia suspensa contains C6 -C2 glucoside conjugates (CCGCs) that are chemically unstable, thereby hindering their isolation and purification. In the present study, ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF) was utilized to screen and identify unstable CCGCs in the fruits and leaves of F. suspensa without any tedious isolation and purified process based on independent information acquisition (also called MSE ) and individual MS/MS experiments. Diagnostic product ion filtering (DPIF) was further applied to mine unknown analogs in MSE high energy levels based on characteristic m/z of key substructures. A modified nomenclature for CCGCs is hereby proposed to facilitate discussions. Possible fragmentation pathways of major types of known CCGCs were proposed and used for deducing their structures. A total of 8 potentially new CCGCs were discovered and initially identified. The accuracy of their identification was further verified by structural elucidation of 3 unstable CCGCs isolated from the fruits of F. suspensa using 1D and 2D-NMR spectroscopy. The established UPLC-QTOF-MSE -based DPIF technique facilitates the rapid discovery and direct identification of unstable CCGCs in fruits and leaves of F. suspensa.