"Nanobiotechnology for health-care"
The influence of Au nanoparticles on metabolic state of cryopreserved mesenchymal stem cells from adipose tissue
N.O. Volkova, M.S. Yukhta, L.V. Sokil, L.V. Stepanyuk, L.G. Chernyshenko, A.M. Goltsev
Institute for Problems of Cryobiology and Cryomedicine, Natl. Acad. of Sci. of Ukraine, Kharkiv
E-mail: volkovana781@gmail.com
The optimum quantity of nanoparticles is defined by the balance between pronounced direct action and low side effects like unwanted cytotoxicity. The influence of Au nanoparticles (AuNPs) on phenotypic characteristics, mitochondrial activity and antioxidant status of cryopreserved mesenchymal stem cells (CrMSCs) from adipose tissue was investigated.
The AuNPs (Sigma-Aldrich, USA) with an initial metal concentration of 45 μg/ml and an average size of 15 nm were used in the study. CrMSCs from adipose tissue were incubated with AuNPs at final concentrations of 4, 6, 10, 20 μg/ml for 1 hour. The control was cells without the influence of AuNPs (0 μg/ml). For phenotypic analysis CrMSCs were stained with CD45-FITC, CD44-FITC, CD73-FITC, CD90-FITC, CD105-PE monoclonal antibodies (BD Biosciences, USA). The functional state of mitochondria was investigated using JC-1 detection kit (BD). Analysis of total antioxidant status (TAS) was performed using Randox test kit (UK). The results were processed with Student's t-test using Excel software.
Adding of AuNPs at studied concentrations did not lead to any significant changes in the level of expression of CD45, CD90, CD73. Using of 10 and 20 µg/ml AuNPs resulted in reduction of CD44+ cells by 12% and 18% respectively compared to the control. Content of CD105+ cells were reduced by 9% in the case of 20 μg/ml AuNPs concentration. The use of 4 and 6 μg/ml AuNPs did not influence mitochondrial activity and TAS in CrMSCs. AuNPs at concentrations of 10 and 20 μg/ml decreased the TAS index (14 and 22% correspondently) and JC-1+ cells content (11% and 18% correspondently) if compare with the control samples. It was found that AuNPs in concentrations of 4 and 6 µg/ml are safe for CrMSCs from adipose tissue, while increase up to 10 µg/ml has a toxic effect manifested by the change of phenotype, mitochondrial and antioxidant activities. 
Thus, the obtained results are related to the field of applied nanotechnology, that extends to clinical medicine, especially in development of addressed drug delivery to target cells or organs.
The work was carried out within the framework of the target research program of the NAS of Ukraine "Genomic, molecular and cellular bases of development of innovative biotechnologies", agreement No. 2.2.6.132/20.