Preparation of cell penetrating peptide TAT-modified liposomes loaded with salvianolic acid B and its effect on proliferation and migration of human skin fibroblasts / 中草药

ABSTRACT

Objective To prepare the liposomes of salvianolic acid B modified with cell penetrating peptide TAT (SAB-TAT-LIP), of which has effects on preventing and treating hypertrophic scars (HS), and establish the method of quality evaluation, as well as preliminarily investigate the effect on the proliferation and migration of human skin fibroblasts (HSF). Methods Liposomes were prepared by pH gradient reverse-phase evaporation method, and the entrapment efficiency was measured by ultrafiltration. Box-Behnken design was performed to optimize the formulation of liposomes by using encapsulation rate as evaluating index. The physicochemical properties of liposomes including morphology, entrapment efficiency, particle size, zeta potential, in vitro release and transdermal absorption, and stability were studied. In addition, the effect of liposomes on proliferation of HSF was examined by MTT assay, and the effect of liposomes on migration of HSF was investigated by scratching method and Transwell assay. Results Based on the optimal formulation of SAB-TAT-LIP, the entrapment efficiency of salvianolic acid B was (86.70 ± 0.85)%, the average particle size was (219.90 ± 5.09) nm, and the zeta potential was (-9.25 ± 0.92). The in vitro 24 h cumulative release was 62.49% of the total drug with no burst effect. The in vitro 32 h cumulated skin penetration rate was 17.21%, the permeance rate was (28.33 ± 4.9) μg/(cm2∙h), and the retention volume of dermis was (44.39 ± 6.87) μg/cm2. The stability was good when placed at 4 ℃ for 10 d. The in vitro cell studies showed that SAB-TAT-LIP can significantly inhibit the proliferation, migration and invasion of human skin fibroblasts, compared with the control group (P < 0.01). Conclusion The optimized SAB-TAT-LIP have higher encapsulation efficiency, smaller particle size, good sustained release effect, and good dermal retention effect which all satisfy the in vitro release and transdermal regulation of local transdermal preparation, and it can significantly inhibit the proliferation, migration and invasion of human skin fibroblasts in vitro.