Comparing the impact of various sucrose concentrations on the performance of various lipid-based nanoformulations for the delivery of trans-resveratrol as a model drug

Abstract

Drug delivery via aerosolization is a non-invasive and ideal method for targeting the pulmonary system, achieving localized effect. This study aims to formulate and compare five different types of lipid-based powder formulations: pro-micelles (F1-F3), pro-niosomes (F4-F6), pro-liposomes (F7-F9), pro-transfersomes (F10-F12), and pro-nanostructured lipid carriers (F13-F15), using three different lipid-to-sucrose carrier ratios (1:05, 1:10, and 1:15 w/w) via a slurry method, using a model anticancer drug, trans-resveratrol (TRES). Aerosolization performance was determined via a two-stage impinger (TSI) paired with two medical nebulizers (i.e., air-jet and vibrating mesh). Post-hydration, vesicles from pro-niosome formulations (F3-F6) demonstrated significantly larger particle size (>1200 nm), while from pro-transfersomes (F11-F13) showed smaller particle size (<125 nm). Entrapment efficiency for all formulations exceeded 90%. An elevated dynamic viscosity was recorded for all formulations when employing a type B Ostwald viscometer as opposed to A, C and D, regardless of formulation type. Upon nebulization, the vibrating mesh nebulizer was associated with an extended nebulization time (23-32 min), short sputtering time (1.57-5.59) and high mass output (79-95%) in compared to air-jet nebulizer (irrespective of formulation type). Both nebulization time and mass output increased significantly with increasing sucrose concentration in formulations (from 1:5 to 1:15 w/w ratio). Upon aerosolization performance, the vibrating mesh showed a significantly higher emitted dose (ED; 72-98%) and fine particle fraction (FPF; 60-81%) than its counterpart nebulizer. The in-vitro release profile depicted zero-order controlled drug release. Moreover, over a period of 24 h, the maximum release of TRES was found at pH 5, rather than at pH 7. Thus, it was concluded that formulations with a higher viscosity (with increasing sucrose concentration) and a vibrating mesh nebulizer are the best combination for peripheral drug deposition.

Introduction

Globally, lung cancer is the second most prevalent malignant cancer with the highest fatality rate of all malignancies [1]. It is broadly categorized histologically into either small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC) (with NSCLC accounting for more than 85% of lung cancer patients globally) [2]. Despite commonly being a first-line treatment, systemic chemotherapy often affects non-cancerous cells, leading to significant side effects and limited effectiveness. In contrast, administering anticancer drugs through inhalation allows high concentrations of chemotherapeutic agents to be delivered directly to the lungs, providing a localized effect that enhances antitumor activity whilst minimizing systemic side effects [3]. Nanoparticles have shown superior performance over traditional dosage forms by improving efficacy, reducing side effects, and increasing stability, thanks to their small size, larger surface area, and precise targeting capabilities [4,5].

Resveratrol (3,5,4-trihydroxystilbene), a natural polyphenol stilbene comprising two phenol rings joined together by ethylene bridges, existing in two isomeric forms, cis- and trans-resveratrol. Antioxidant, anti-inflammatory, cardioprotective, and cancer-fighting actions are some of the benefits associated with the phytoestrogen trans-resveratrol (TRES). The trans-form is more biologically active, but less stable, as it converts to the cis-form when exposed to UV light and at higher pH levels [6]. Additionally, TRES undergoes extensive liver metabolism and has poor water solubility, leading to low bioavailability [7,8]. This compound has acquired considerable attention related to its chemopreventive and anticancer properties. Comprehensive in-vitro and in-vivo research has shown that TRES can disrupt all phases of carcinogenesis, encompassing tumour initiation, development, and advancement, while also inhibiting angiogenesis and metastasis [9]. It exerts its effects by altering many signalling pathways associated with cell proliferation, inflammation, apoptosis, and tumour formation, including the Akt, NF-κB, and MAPK pathways [9]. Furthermore, recent studies have demonstrated its capacity to induce apoptosis, inhibit metastasis, and modulate the tumour microenvironment across multiple cancer types [10]. Although TRES is not a traditional chemotherapeutic agent, the recognised anticancer properties of TRES endorse its application as a model bioactive chemical in cancer research.

Previous studies have shown that transfersomes and liposomes face stability concerns due to issues such as aggregation, fusion, and leaking of drugs from the vesicles, attributed to the phospholipid component [11]. Similar instability has been observed in aqueous suspensions of niosomes and micelles [12]. To address these problems, lipid-based nanoformulations have been developed in powder form using carbohydrate carriers such as sucrose and lactose [[13], [14], [15]]. These carriers act as dispersing agents and emulsifiers, aiding in the breakdown of lipid-based vesicles into smaller, more stable particles by preventing phase separation and sedimentation. Additionally, carbohydrate carriers enhance viscosity and modify the rheological properties of lipid-based formulations. Therefore, pro-lipid-based dry powders are superior to lipid-based suspensions due to their enhanced stability, higher drug loading capacity, controlled and sustained drug release, reduced risk of aggregation, longer shelf life, and ease of reconstitution [16].

Aerosol production suitable for pulmonary delivery is heavily influenced by the interplay between the working mechanism of various nebulizers and the physicochemical parameters of the formulations, such as pH, viscosity, and surface tension [6,17]. In the case of pulmonary disease conditions, nebulized vesicles improve the residence time of drugs in the lungs, potentially improving therapeutic impact, without causing substantial systemic adverse effects [18]. To study the particle deposition, the British Pharmacopeia recommends using an artificial lung model called the two-stage or twin impinger (TSI). The TSI, with upper and lower stages and a cut-off diameter of 6.4 μm between them, simulates the upper and lower respiratory tracts to measure drug deposition in the lungs. Formulation deposition in the lungs occurs through three main mechanisms: inertial impaction, sedimentation, and diffusion, all of which depend on the particle aerodynamic diameter [19].

In this study, several lipid-based formulations (pro-micelles, pro-niosomes, pro-liposomes, pro-transfersomes, and pro-nanostructured lipid carriers (pro-NLCs)) were investigated in order to assess their viability as reconstituted dry powder nanoformulations for pulmonary administration through nebulization. These various formulations correspond to different classes with distinct structural characteristics, including surfactant micelles, non-ionic surfactant vesicles, phospholipid vesicles, and lipid matrix nanoparticles [[20], [21], [22], [23], [24], [25], [26], [27]]. Due to these structural differences, the formulations behave differently during dehydration (i.e., in a rotary evaporator), storage, and rehydration. Therefore, investigating multiple lipid-based pro-powder systems facilitates a systematic evaluation of their ability to form stable dry formulations and reconstituted nanoscale dispersions upon hydration suitable for nebulization [[28], [29], [30]], making them suitable for nebulization. Moreover, excipients for the dry-lipid-based powder formulations were chosen based on their proven and defined functional roles in creating distinct nanocarrier systems. Pro-micelles (F1–F3) were formulated utilizing amphiphilic surfactants (e.g., Tween 80) in order to facilitate micelle formation. Pro-niosomes (F4–F6) consisted of non-ionic surfactants including Tween 80 or Span 60 with cholesterol for bilayer formation. Pro-liposomes (F7–F9) employed phospholipids (e.g., soy or egg lecithin) with or without cholesterol to create stable vesicles. Pro-transfersomes (F10–F12) included phospholipids with edge activators (Span 80 or Tween 80) to enhance membrane flexibility. Pro-nanostructured lipid carriers (F13–F15) were prepared using a mixture of liquid and solid lipids along with an edge activator to enhance drug accommodation. Sucrose, as a carbohydrate sugar, was employed as a hydrophilic carrier due to its high solubility as well as suitability for producing free-flowing powders [31,32]. Various lipid-to-sucrose ratios (1:5, 1:10, and 1:15 w/w) were explored to assess their effect on formulation properties as well as aerosol generation and deposition in the pulmonary system. In this study, formulations were characterized based on particle size, drug entrapment, viscosity, and in-vitro drug release. The performance of nebulizers was assessed via deposition of a drug in the TSI. The aim was to explore how the viscosity of each formulation influenced nebulization performance, specifically nebulization time, sputtering time, and mass output, using two types of nebulizers: air-jet and vibrating mesh. Finally, the deposition of TRES in the nebulizer reservoir and the stages of the TSI were analyzed to determine the most effective nebulizer and formulation type.

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Materials

Soya phosphatidylcholine (SPC; Lipoid S-100) was purchased from Lipoid, Switzerland. Trans-resveratrol (TRES) was obtained from Manchester Organics Ltd., UK. Propylene glycol dicaprylate (PGD; Miglyol 840) was generously gifted by IOI Oleochemicals, Witten, UK. Glycerol dibehenate (GDB; Compritol 888 ATO) was a gift from Gattefosse, France. Tween 80 and cholesterol were acquired from Sigma Aldrich, UK. HPLC grade acetonitrile, analytical grade sucrose, formic acid, acetic acid, absolute ethanol, and tetrahydrofuran were bought from Fischer Scientific, UK. Spectra/Por 7 pre-treated dialysis membrane (MWCO: 3.5 kDa) was procured from Spectrum Laboratories, USA.

Anila Mathew Thevarkattil, Kajanthan Balasundaram, Nozad Hussein, Huner Omer, Sakib Yousaf, Chahinez Houacine, Adeeb Shehzad, Ruba Bnyan, Abdelbary Elhissi, Iftikhar Khan, Comparing the impact of various sucrose concentrations on the performance of various lipid-based nanoformulations for the delivery of trans-resveratrol as a model drug, Journal of Drug Delivery Science and Technology, Volume 125, 2026, 108671, ISSN 1773-2247, https://doi.org/10.1016/j.jddst.2026.108671.