Potent Melanin Production Enhancement of Human Tyrosinase Gene by Tat and an Entrapment in Elastic Cationic Niosomes: Potential Application in Vitiligo Gene Therapy

Authors

  • Jiradej Manosroi,

    Corresponding author
    1. Faculty of Pharmacy, Chiang Mai University, Chiang Mai 50200, Thailand
    2. Natural Products Research and Development Center (NPRDC), Science and Technology Research Institute (STRI)/Faculty of Pharmacy, Chiang Mai University, Chiang Mai 50200, Thailand
    Search for more papers by this author
  • Narinthorn Khositsuntiwong,

    1. Faculty of Pharmacy, Chiang Mai University, Chiang Mai 50200, Thailand
    Search for more papers by this author
  • Friedrich Götz,

    1. Department of Microbial Genetics, Faculty of Biology, University of Tübingen, Tübingen, Germany
    Search for more papers by this author
  • Rolf G. Werner,

    1. Boehringer Ingelheim Company, Ingelheim am Rhein, Germany
    Search for more papers by this author
  • Worapaka Manosroi,

    1. Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand
    Search for more papers by this author
  • Aranya Manosroi

    1. Faculty of Pharmacy, Chiang Mai University, Chiang Mai 50200, Thailand
    2. Natural Products Research and Development Center (NPRDC), Science and Technology Research Institute (STRI)/Faculty of Pharmacy, Chiang Mai University, Chiang Mai 50200, Thailand
    Search for more papers by this author

Corresponding author: Jiradej Manosroi, jiradej.manosroi8@gmail.com, pmpti006@chiangmai.ac.th

Abstract

Potent melanin production enhancement of human tyrosinase plasmid (pAH7/Tyr, P) in mouse melanoma cells (B16F10) by Tat peptide (T) and an entrapment in elastic cationic niosomes (E) was described. The E composed of Tween 61/cholesterol/dodecyl dimethyl ammonium bromide at 1:1:0.5 molar ratio was prepared by freeze-dried emptying liposomes method. PE at P/E ratio of 1:160 w/w and TPE at T/P/E ratio of 0.125:1:160, 0.25:1:160, and 0.5:1:160 w/w/w were prepared. The final concentration of the plasmid in the study was 4 ng/μL. By sulforhodamine B assay, PE and TPE complexes showed slight or no cytotoxic effect. The cells transfected with TPE (0.5:1:160) exhibited the highest enhancement of tyrosinase enzyme activity of 11.82-, 7.67-, 5.07-, and 6.29-folds of control, P, PE, and TP (0.5:1) and melanin production of 13.03-, 8.46-, 5.36-, and 6.58-folds of control, P, PE, and TP (0.5:1), respectively. The elastic cationic niosomes demonstrated an increase in thermal stability of P at 4 ± 2, 25 ± 2, and 45 ± 2 °C. The vesicular size and the zeta potential values of PE and TPE complexes were slightly increased but still in the range of stable dispersion (out of ±30 mV). These results indicated the high potential application of the TPE complexes for further investigation for vitiligo gene therapy.

Ancillary