Enhanced transdermal lymphatic drug delivery of hyaluronic acid modified transfersome for tumor metastasis therapy.

Materials Sodium form of hyaluronic acid (HA, MW, 10 kDa) was a gift of Kolon Life Science, Korea. EDC.HCl (99%) and N-Hydroxy succinimide (NHS, 99%) were purchased from Shanghai Medeep Co., Ltd. Monostearin (glycerol α-monostearate, GMS), Sodium deoxycholate (DOC) and fluorescein isothiocyanate (FITC) were purchased from Solarbio (Beijing, China). Doxorubicin hydrochloride (DOX) was supplied by Zhejiang Hai zheng Co. Ltd. (China). All other reagents and solvents were of analytical grade. Preparation and characterization of hyaluronic acid modified transfersome (HA-GMS-T) Amphiphilic HA-GMS were synthesized using a previous method. [1] HA-GMS was labeled by FITC or rhodamine for fluorescence identification, respectively. [1,2] Transfersomes were prepared through lipid film method. Briefly, 70 mg lecithin was dissolved in chloroform/ethanol (1:1, v/v) and evaporated to dry to form lipid film under reduced pressure. Then, the transfersomes were produced by hydration of the lipid film with 10 mM PBS (pH 7.4) containing 25 mg DOC at 55 ℃ for 30 min, and size reduction with sonication. HA-GMS was added during hydration with final concentration of 1 mg/mL to give HA modified transfersome (HA-GMS-T). Various parameters including lecition/DOC ratio, hydration temperature, duration and medium, pH condition and solvent were screened out to optimize the preparation (Table S1). Size analysis and zeta potential were determined on a Zetasizer ZEN 3600 Nano Series apparatus (ZEN, UK). The morphology of HA-GMS-T was observed via TEM (JEM-1200EX JEOL Ltd., Japan) and CLSM (Carl-Zeiss, German) at Ex 543 nm. Transmittance at 600 nm of HA-GMS-T was determined to evaluate its stability.