This study introduces and statistically optimizes novel Tetra-Synergistic Bilosomes (TSB), an advanced nano-platform for topical delivery of voriconazole (VOR) in otomycosis therapy. The term “Tetra-Synergistic” reflects the incorporation of four complementary components: Cremophor® EL, sodium deoxycholate, Brij® 58, and Brij® S2, working in synergy to enhance therapeutic efficacy. TSB was fabricated using the thin-film hydration method and optimized through a 2 ³ D-optimal design. Three formulation factors were investigated: Cremophor® EL: VOR ratio, sodium deoxycholate: cholesterol ratio, and Brij® 58: Brij® S2 ratio, with the optimization goal of maximizing EE%, minimizing vesicular size, and maintaining ZP within the acceptable stability range. The model demonstrated excellent predictability, achieving a desirability value of 0.888. The optimized TSB exhibited high EE% (91.6%), a suitably small VS (217.9nm), and stable ZP (− 24.1mV). FTIR confirmed VOR entrapment, while TEM revealed nano-sized spherical vesicles. Moreover, TSB displayed a biphasic In-vitro release profile and remained stable for three months under refrigeration. Importantly, microbiological evaluation confirmed the superior antifungal performance of TSB. Compared with VOR suspension, TSB showed a larger inhibition zone, a lower minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC), and a faster fungicidal effect, eradicating fungal growth in 33% less time. Ex-vivo permeation studies revealed a 2.18-fold enhancement ratio, supported by confocal laser scanning microscopy confirming deeper tissue penetration. Histopathological analysis further validated the safety of topical application without any tissue damage. Collectively, these results establish TSB as a pioneering, microbiologically potent, and clinically promising nano-vesicular system for the effective management of otomycosis.

This work aimed to prepare Terconazole loaded proniosomes (TCZ-PNS) utilizing modified coacervation technique for the management of fungal keratitis. Terconazole (TCZ) is a potent antifungal with poor aqueous solubility posing intricacies in its incorporation in ocular formulations. A 2³ factorial design was adopted to probe independent formulation variables including A: Lecithin: cholesterol ratio, B: Surfactant: cholesterol ratio and C: Span® 80 contribution (% of total SAA). The formulae, generated by the design, were prepared and scrutinized regarding entrapment efficiency (%EE), particle size (PS), polydispersity index (PDI) and zeta potential (ZP). Numerical desirability algorithms selected an optimum TCZ-PNS which boasted plausible %EE (89.51 % ± 0.94 %), nanoscale vesicles consistent with TEM measurements (247.9 ± 0.42 nm), a sufficiently high ZP (-43.42 ± 0.85 mV), and an in-vitro biphasic release profile that remained stable even after Gamma irradiation and short-term storage. The transcorneal ex-vivo permeation of TCZ-PNS was higher than that of TCZ suspension (≈ 2-fold). The formulation was further evaluated for pH, corneal hydration threshold, and histopathological safety, confirming its suitability for ocular application. Confocal laser microscopy revealed substantial corneal uptake (approximately twice as deep as of TCZ suspension). Additionally, microbiological assessments of the optimal TCZ-PNS compared to TCZ suspension demonstrated an inhibition zone nearly 50 % larger, a significantly lower MIC and MFC (64-fold reduction), and enhanced biofilm inhibition activity across most tested concentrations. These findings suggest that TCZ-PNS could be a propitious treatment choice to deeply deliver antifungal therapy for the eradication of deeply rooted and inaccessible fungal keratitis.

Herein, we investigated the preparation and characterization of Terconazole loaded edge-activated hybrid elastosome (TCN-EHE) adopting thin film hydration technique for the treatment of ocular mycosis. Terconazole (TCN) is a broad spectrum antimycotic agent suffering from sparse aqueous solubility impeding its use in ophthalmic preparations. The scrutinized formulation variables namely X₁: Surfactant: Edge activator ratio (SAA: EA), X₂: Pluronic® L121 contribution (% of total SAA) and X₃: EA concentration (%w/v) were optimized adopting D-optimal design. Ten runs were prepared and characterized regarding their entrapment efficiency, particle size, polydispersity index and zeta potential. An optimized formula was generated, with high desirability, exhibited satisfactory entrapment efficiency, nanoscaled particle size aligning with TEM, plausible zeta potential and bi-phasic release pattern which were not altered after short-term storage. The optimized TCN-EHE displayed 1.94-fold enhanced ex-vivo corneal permeation flux. Safety was ratified through measured corneal hydration level, pH and histopathological evaluation. In-vivo corneal uptake visualized by confocal laser microscopy demonstrated 2.7-fold deeper penetration. Moreover, Superior antifungal activity has been demonstrated displaying 37 % bigger zone of inhibition, 8-fold lower minimum inhibitory and minimum fungal concentration alongside significantly higher biofilm inhibition activity at all tested concentrations for the optimized TCN-EHE compared to TCN suspension. Conclusively, we could prospect that TCN-EHE might be a revamped therapeutic alternative for the delivery of poorly soluble antimycotic agents for the combat of ocular mycosis.

Purpose: Development of hyaluronic acid conjugated metformin-phospholipid sonocomplexes (HA-MPS), a biphasic complexation product compiled for enhancing both the lipophilicity and targeting potential of Metformin (MET) to CD44 receptors on pancreatic cancer. Methods: MET was chemically conjugated to hyaluronic acid (HA) via amide coupling reaction. Then, the HA conjugated MET was physically conjugated to Lipoid™S100 via ultrasound irradiation. A combined D-optimal design was implemented to statistically optimize formulation variables. The HA-MPS were characterized through solubility studies, partition coefficient, drug content uniformity, particle size and zeta potential. The optimized HA-MPS was tested via proton nuclear magnetic resonance, infrared spectroscopy to elucidate the nature of physicochemical interactions in the complex which was further scrutinized on molecular level via molecular docking and dynamic simulation. Results: The solubility and partition studies showed a lipophilicity enhancement up to 67 folds as they adopted inverted micelles configuration based on the packing parameter hypothesis. The optimized HA-MPS showed 11.5 folds lower IC₅₀, extra 25% reduction in oxygen consumption rate, better reduction in hypoxia-inducible factor and reactive oxygen species in MiaPaCa-2 cells. Conclusion: These results proved better internalization of MET which was reflected by abolishing hypoxic tumour microenvironment, a mainstay toward a normoxic and less resistant pancreatic cancer.