"Nanochemistry and biotechnology"

Change in functional state of mesenchymal stromal cells after incubation with silver nanoparticles

N.A. Volkova, E.V. Pavlovich, A.N. Goltsev
Institute for Problems of Cryobiology and Сryomedicine of the Natl. Acad. of Sci. of Ukraine, Pereyaslavskaya str. 23, Kharkov-61015, Ukraine.
E-mail: lenapavlovich@gmail.com

Due to its unique physical and chemical properties nanoscale materials have a dose-dependent effect on biological objects. Finding the presence / absence of cytotoxic action of silver nanoparticles (AgNPs) is an important direction of nanobiotechnological research. The effect of AgNPs on functional state of bone marrow mesenchymal stromal cells (MSCs) was investigated.
The AgNPs (Sigma-Aldrich, USA) were obtained by citrate synthesis with an initial metal concentration of 20 μg/mL. The average size of AgNPs was 40 nm. The cells were incubated in nutritive medium supplemented with AgNPs in concentrations of 2, 4, 6, 10 μg/mL (1 hour at 37 ºC). For immunophenotypic analysis the MSCs were stained with CD45-FITC, CD44-FITC, CD73-FITC, CD90-FITC, CD105-PE monoclonal antibodies (BD Biosciences, USA). The apoptotic/necrotic processes (Annexin V/7AAD, BD Biosciences, USA) and mitochondrial state (JC-1, BD Biosciences, USA) in the cells were investigated with FACS Calibur. The group of comparison (control) was MSCs without AgNPs. The results were processed with Student's t-test using Excel software.
Adding AgNPs of the concentrations studied did not lead to any significant alteration in the level of expression of CD 45, CD 29, CD 105. In contrast, the AgNPs concentration of 10 μg/mL resulted in a reduction of CD 44 and CD 90 cells by 4.92% and 8.19% respectively. The results obtained by flow cytometry suggested that AgNPs at concentrations of 2 and 4 μg/mL on the 1 hour of incubating did not cause the development of necrosis/apoptosis processes and alteration of mitochondrial activity in MSCs. The use of AgNPs at concentration of 6 and 10 μg/mL a decrease of mitochondrial activity, percentage of Аnnexin V-/7AAD- cells and an increase of the number of apoptotic cells were observed compared with the control MSCs samples. The obtained results are related to the field of applied nanotechnology, which extends to clinical medicine, especially in development of addressed drug delivery to target cells or organs.
This work has been supported by project No 70/18-Н NAS of Ukraine "Biosafety and efficiency of interaction of inert metal nanoparticles with mesenchymal stem cells of bone marrow".