Integrating Network Pharmacology and Molecular Docking to Analyse the Potential Mechanism of action of Macleaya cordata (Willd.) R. Br. in the Treatment of Bovine Hoof Disease

Based on network pharmacological analysis and molecular docking techniques, the main components of M. cordata for the treatment of bovine relevant active compounds in M. cordata were searched for through previous research bases and literature databases, and then screened to identify candidate compounds based on physicochemical properties, pharmacokinetic parameters, bioavailability, and drug-like criteria. Target genes associated with hoof disease were obtained from the GeneCards database. Compound−target, compound−target−pathway−disease visualization networks, and protein−protein interaction (PPI) networks were constructed by Cytoscape. Gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed in R language. Molecular docking analysis was done using AutoDockTools. The visual network analysis showed that four active compounds, sanguinarine, chelerythrine, allocryptopine and protopine, were associated with the 10 target genes/proteins (SRC, MAPK3, MTOR, ESR1, PIK3CA, BCL2L1, JAK2, GSK3B, MAPK1, and AR) obtained from the screen. The enrichment analysis indicated that the cAMP, PI3K-Akt, and ErbB signaling pathways may be key signaling pathways in network pharmacology. The molecular docking results showed that sanguinarine, chelerythrine, allocryptopine, and protopine bound well to MAPK3 and JAK2. A comprehensive bioinformatics-based network topology strategy and molecular docking study has elucidated the multi-component synergistic mechanism of action of M. cordata in the treatment of bovine hoof disease, offering the possibility of developing M. cordata as a new source of drugs for hoof disease treatment.

Improving Transungual Permeation Study Design by Increased Bovine Hoof Membrane Thickness and Subsequent Infection

Ungual formulations are regularly tested using human nails or animal surrogates in Franz diffusion cell experiments. Membranes sometimes less than 100 µm thick are used, disregarding the higher physiological thickness of human nails and possible fungal infection. In this study, bovine hoof membranes, healthy or infected with Trichophyton rubrum, underwent different imaging techniques highlighting that continuous pores traversed the entire membrane and infection resulted in fungal growth, both superficial, as well as in the membrane’s matrix. These membrane characteristics resulted in substantial differences in the permeation of the antifungal model substance bifonazole, depending on the dosage forms. Increasing the thickness of healthy membranes from 100 µm to 400 µm disproportionally reduced the permeated amount of bifonazole from the liquid and semisolid forms and allowed for a more pronounced assessment of the effects by excipients, such as urea as the permeation enhancer. Similarly, an infection of 400-µm membranes drastically increased the permeated amount. Therefore, the thickness and infection statuses of the membranes in the permeation experiments were essential for a differential readout, and standardized formulation-dependent experimental setups would be highly beneficial.

Nanoindentation and Hierarchy Structure of the Bovine Hoof Wall

The bovine hoof wall with an α-keratin structure protects the bovine foot from impact loads when the cattle are running. Reduced modulus, hardness and creep behavior of the bovine hoof wall have been investigated by a nanoindentation technique. The average reduced modulus of the Transverse Direction (TD) specimens from the outside to inside wall is 3.76 and 2.05 GPa, respectively, while the average reduced modulus of the Longitudinal Direction (LD) specimens from the outside to inside wall is 4.54 and 3.22 GPa, respectively. Obviously, the orientation and the position of the bovine hoof wall have a significant influence on its mechanical properties. The use of the generalized Voigt–Kelvin model can make a good prediction of creep stage. Mechanical properties of the LD specimens are stronger than those of the TD specimens. The bovine hoof wall has a layered structure, which can effectively absorb the energy released by the crack propagation and passivate the crack tip. Therefore, a kind of structural model was designed and fabricated by three-dimensional printing technology, which has a 55% performance improvement on fracture toughness. It is believed that the reported results can be useful in the design of new bionic structure materials which may be used in motorcycle helmets and athletes’ protective equipment to achieve light weight and improved strength at the same time.

Optimization of Drug Permeation from 8% Ciclopirox Cyclodextrin/Poloxamer-Soluble Polypseudorotaxane-Based Nail Lacquers

Cyclodextrin/poloxamer-soluble polypseudorotaxane-based nail lacquers have demonstrated significant capacity for promoting the permeation of drugs into the nail plate. Furthermore, previous studies have shown that the use of hydroalcoholic blends as vehicles promotes drug permeation. The work described herein studies the effect of the type of alcohol used in the lacquer preparation, and the composition of the vehicle is optimized to obtain soluble doses of 8% and to promote the diffusion of ciclopirox base and olamine across the nail. Permeation studies on different types of alcohols show that optimum results are achieved with short-chain alcohols, and that results become less satisfactory the higher the number of alcohol carbons. In addition, solubility and penetration studies on the bovine hoof have enabled the composition of the lacquer to be optimized for both forms of ciclopirox. The results suggest that optimized lacquers have better ciclopirox diffusion and penetration properties than the commercial reference lacquer. Lastly, in vivo studies in which optimized ciclopirox olamine lacquer was applied for 45 days to the nails of healthy volunteers showed that it caused no negative effects or changes to the nail surface. These results demonstrate the significant potential of cyclodextrin/poloxamer-soluble polypseudorotaxane-based nail lacquers for the ungual administration of drugs.

Development and Validation of an HPLC–UV Method to Quantify Tavaborole During in Vitro Transungual Permeation Studies

A selective and rapid reversed-phase HPLC–UV method was developed and validated to quantify tavaborole (TAV; AN2690) in biological samples, i.e., in receiving phase and in bovine hoof membrane extract derived from in vitro transungual permeation studies. A simple solid–liquid extraction procedure was used to recover the drug from the bovine hoof slices. TAV chromatographic separation was achieved on a Luna PFP column (150 × 4.6 mm, 5 μm) using a mobile phase consisting of a 70% phosphoric acid solution (10 mM, pH 2.0) with 30% acetonitrile. The detection wavelength was set to 220 nm using a UV detector. The method exhibited good linearity in the calibration ranges, which were 0.5–8.0 and 0.03–2.5 μg/mL for the receiving phase and hoof membranes, respectively. The obtained LOD and LOQ values were 0.023 and 0.069 μg/mL, respectively, for the receiving phase and 0.0024 and 0.007 μg/mL for the bovine hoof membrane extracts. In all cases, the CV for intraday and interday precision was widely below the limit of 2%, demonstrating good precision. The analytical method described was sensitive, precise, linear, and accurate and could be applicable for clinical and bioanalytical studies as an alternative to other analytical methods, which are quite expensive and not always available in research laboratories.