Evaluation of the localization and biological effects of PAMAM G3 dendrimer-biotin/pyridoxal conjugate as HaCaT keratinocyte targeted nanocarrier

Magdalena Szuster; Łukasz Stanisław Uram; Aleksandra Filipowicz-Rachwał; Stanisław Wołowiec; Elżbieta Wałajtys-Rode

doi:10.18388/abp.2018_2767

Magdalena Szuster Faculty of Chemistry, Rzeszow University of Technology, Rzeszów, Poland
Łukasz Stanisław Uram Faculty of Chemistry, Rzeszow University of Technology, Rzeszów, Poland
Aleksandra Filipowicz-Rachwał Faculty of Medical Sciences, Rzeszow University of Information Technology and Management, Rzeszów, Poland
Stanisław Wołowiec Centre for Innovative Research in Medical and Natural Sciences, Faculty of Medicine, University of Rzeszow, Rzeszów, Poland
Elżbieta Wałajtys-Rode Department of Drug Technology and Biotechnology, Faculty of Chemistry, Warsaw University of Technology, Warsaw, Poland

DOI: https://doi.org/10.18388/abp.2018_2767

Abstract

Recognition of the molecular mechanisms of keratinocyte participation in normal skin homeostasis and in pathogenesis may lead to creation of more effective tools for topical application of cosmetics, cosmeceutics and drugs to a particular location within the skin for prevention and therapy of many skin disorders and diseases. For this purpose, the PAMAM G3 dendrimer with amide linkages of 9 biotin molecules and 10 molecules of pyridoxal phosphate (BC-PAMAM) was constructed, and its biological properties and cellular uptake and localization were investigated in the HaCaT keratinocytes. BC-PAMAM is nontoxic for HaCaT cells, as estimated by two assays (Neutral Red and tetrazolium salt reduction, XTT), and revealed low apoptosis induction at up to 50 µM concentration. Fluorescent labeled BC-PAMAM accumulates in HaCaT cells with high efficiency in a concentration–dependent manner. Its mitochondrial localization, estimated as Mander’s colocalization coefficient, is substantially lower than the native PAMAM, and that correlates with its cytotoxicity. The only undesirable, but significant inhibitory effect on cell mobility, evaluated by the wound healing test, was observed at 10 µM BC-PAMAM. The important anti-inflammatory action of BC-PAMAM was clearly documented by decreased production of total IL-1α, assayed with an ELISA test with unstimulated and stimulated by bacterial antigens (LPS and GroEL) HaCaT cells. Thus, it is expected that the biotin pyridoxal phosphate conjugated PAMAM may be considered as a potential carrier for safe delivery of vitamins and drugs into the epidermis.