In Vitro TyRP-1 Knockdown Based on siRNA Carried by Liquid Crystalline Nanodispersions: an Alternative Approach for Topical Treatment of Vitiligo
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Vitiligo is a skin disease characterized by depigmentation and the presence of white patches that are associated with the loss of melanocytes. The most common explanation for the cause of this condition is that it is an autoimmune condition. TyRP-1 is involved in melanin pigment synthesis but can also function as a melanocyte differentiation antigen. This protein plays a role in the autoimmune destruction of melanocytes, which results in the depigmentation, characteristic of this disease. In this study, we evaluated liquid crystalline nanodispersions as non-viral vectors to deliver siRNA-TyRP-1 as an alternative for topical treatment of vitiligo.
Liquid crystalline nanodispersions were obtained and characterized with respect to their physical-chemical parameters including size, PdI and zeta potential, as well as Small Angle X-ray Scattering and complexing to siRNA. The effects of the liquid crystalline nanodispersions on the cellular viability, cell uptake and levels of the knockdown target TyRP-1 were evaluated in melan-A cells after 24 h of treatment.
The liquid crystalline nanodispersions demonstrated adequate physical-chemical parameters including nanometer size and a PdI below 0.38. These systems promoted a high rate of cell uptake and an impressive TyRP-1 target knockdown (> 80%) associated with suitable loading of TyRp-1 siRNA.
We demonstrated that the liquid crystalline nanodispersions showed promising alternative for the topical treatment of vitiligo due to their physical parameters and ability in knockdown the target protein involved with autoimmune destruction of melanocytes.
KEY WORDSin vitro knockdown liquid crystalline nanodispersions siRNA TyRP-1 vitiligo
ACKNOWLEDGMENTS AND DISCLOSURES
We thank José Orestes Del Ciampo for technical assistance in the DLS analyses. We also thank Camila Cristina de Oliveira Menezes Bonaldo, Laboratory of Flow Cytometry, Blood Regional Center-FMRP-USP, Ribeirão Preto, São Paulo, Brazil, for the assistance with the flow cytometry. Thanks are due to LNLS, Brazil for the use of the D02A-SAXS2 beamline. The melan-A non-tumorigenic melanocyte cell line was kindly provided by Dr. Marcelo Dias Baruffi from School of Pharmaceutical Science of Ribeirao Preto, University Sao Paulo, Sao Paulo, Brazil. Part of this work was developed within the framework of National Institute of Science and Technology of Pharmaceutical Nanotechnology (INCT-Nanofarma), which is supported by “Fundação de Amparo à Pesquisa do Estado de São Paulo” (Fapesp, Brazil, grant #2014/50928-2) and “Conselho Nacional de Pesquisa” (CNPQ, Brazil, grant #465687/2014-8). L.B. Tofani was the recipient of a CNPq fellowship (grant 133839/2013-3) and a FAPESP fellowship (grant # 2013/06559-0).
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