112
ISSN 1990-519X, Cell and Tissue Biology, 2018, Vol. 12, No. 2, pp. 112–119. © Pleiades Publishing, Ltd., 2018.
Original Russian Text © L.S. Litvinova, V.V. Shupletsova, O.G. Khaziakhmatova, K.A. Yurova, V.V. Malashchenko, E.S. Melashchenko, N.M. Todosenko, M.Yu. Khlusova,
Yu.P. Sharkeev, E.G. Komarova, M.B. Sedelnikova, E.O. Shunkin, I.A. Khlusov, 2017, published in Tsitologiya, 2017, Vol. 59, No. 12, pp. 858–866.
Behavioral Changes of Multipotent Mesenchymal Stromal Cells
in Contact with Synthetic Calcium Phosphates in vitro
L. S. Litvinova
a,
*, V. V. Shupletsova
a
, O. G. Khaziakhmatova
a
, K. A. Yurova
a
, V. V. Malashchenko
a
,
E. S. Melashchenko
a
, N. M. Todosenko
a
, M. Yu. Khlusova
b
, Yu. P. Sharkeev
c, d
, E. G. Komarova
d
,
M. B. Sedelnikova
d
, E. O. Shunkin
a
, and I. A. Khlusov
a, e
a
Laboratory of Immunology and Cell Biotechnology, Immanuel Kant Baltic Federal University, Kaliningrad, 236041 Russia
b
Pathophysiology Department, Siberian State Medical University, Tomsk, 634050 Russia
c
Experimental Physics Department, Tomsk Polytechnic University, Tomsk, 634050 Russia
d
Institute of Strength Physics and Materials Science of the Siberian Branch of the Russian Academy of Sciences,
Tomsk, 634055 Russia
e
Morphology and General Pathology Department, Siberian State Medical University, Tomsk, 634050 Russia
*e-mail: larisalitvinova@yandex.ru
Received July 7, 2017
Abstract—Migration, proliferation, and osteogenic differentiation of human adipose-derived (AD) multipo-
tent mesenchymal stromal cells (MMSCs) during in vitro modeling of indirect contact with calcium phos-
phate (CP) or nanoparticles of synthetic hydroxyapatite (HA) have been studied. The results were registered
with electrode (real-time cell analysis, RTCA) or visual (Cell-IQ) systems of long-term observation of
cell cultures. Bulk specimens were use in a Cell-IQ® v2 MLF device as pure titanium substrates (10 × 10 ×
1 mm
3
) covered by a CP relief (roughness index R
a
= 2.4–4.4 μm) bilateral coating that was prepared by the
micr-arc method from an aqueous solution of orthophosphoric acid (20 wt %), calcium carbonate (9 wt %),
and synthetic HA (6 wt %). HA crystallites (1 mg/mL) were fabricated by mechanochemical synthesis and
served as an irritant in RTCA investigation. The Cell-IQ system identified a 3.5- to 10-fold decrease in cell
number at the interface with CP coatings with differing roughness during 14-day cell culturing. After 21 days,
it was accompanied by a weak reduction of MMSC antigen expression (CD73, CD90, and CD105) as
opposed to an increase in MMSC osteogenic differentiation and intercellular-matrix mineralization. In turn,
HA nanodispersion reduced the speed of MMSC migration by 1.5 times (P < 0.001) during 25-h RTCA
recording, which simulated cell invasion through the microporous membrane (8-μm diameter). Inhibition of
migration and cell division with increased osteogenic differentiation of MMSCs has been suggested to be a
possible effect of biodegradation products of synthetic CP materials.
Keywords: stromal cells of human adipose tissue, motility, trafficking, membrane markers, osteogenic differ-
entiation, hydroxylapatite, calcium phosphate coating, microtopography
DOI: 10.1134/S1990519X18020062
INTRODUCTION
Real-time cell analysis (Atienza et al., 2006) and
Cell-IQ (Meindl et al., 2013) are contemporary tools
for noninvasive investigation and continuous cell
monitoring. They allow real-time determination of the
cell morphology and behavior (viability, migration
adhesion, proliferation, differentiation, maturation)
under normal conditions and after treatment with var-
ious stimuli (pharmacological agents, biomolecules,
viruses, etc.) (Teng et al., 2013).
Multipotent mesenchymal stromal cells
(MMSCs), which are able to migrate into inflamma-
tion, regeneration, and tumor-growth foci and transfer
pharmaceutical drugs, biological molecules, and
nanoparticles (Lepik et al., 2016), have been widely
used in different studies. However, only isolated works
have been devoted so far to modeling and determining
the behavior of MMSCs in real time (Moniri et al.,
2015). The applicability of survey tools to cell interac-
tion with extracellular-matrix components and artifi-
cial-implant materials remains largely untapped (e.g.,
Ozturk et al., 2014).
The bone–implant boundary is a critical link for
reparative bone regeneration. A major approach to
promoting osseointegration of implanted materials is
the use of calcium-phosphate (CP) coating. Depend-
Abbreviations: HA—hydroxyapatite, CP—calcium phosphate,
MI—cell-migration index, AT—adipose tissue, MMSC—multi-
potent mesenchymal stromal cell, EDTA—ethylenedi-
aminetetraacetic acid, RTCA—real-time cell analysis.