The subject of the invention is superparamagnetic nanoparticle probes
based on iron oxides, to advantage magnetite or maghemite, with modified
surface, coated with mono-, di- or polysaccharides from the group
including D-arabinose, D-glucose, D-galactose, D-mannose, lactose,
maltose, dextrans and dextrins, or with amino acids or poly(amino acid)s
from the group including alanine, glycine, glutamine, asparagine,
histidine, arginine, L-lysine, aspartic and glutamic acid or with
synthetic polymers based on (meth)acrylic acid and their derivatives
selected from the group containing poly(N,N-dimethylacrylamide),
poly(N,N-dimethylmethacrylamide), poly(N,N-diethylacrylamide),
poly(N,N-diethylmethacrylamide), poly(N-isopropylacrylamide),
poly(N-isopropylmethacrylamide), which form a colloid consisting of
particles with narrow distribution with polydispersity index smaller than
1.3, the average size of which amounts to 0.5-30 nm, to advantage 1-10
nm, the iron content is 70-99.9 wt. %, to advantage 90 wt. %, the
modification agent content 0.1-30 wt. %, to advantage 10 wt. %.
The particles of size smaller than 2 nm with polydispersity index smaller
than 1.1 can be obtained by a modified method of preparation.
Superparamagnetic nanoparticle probes according to the invention are
prepared by pre-precipitation of colloidal Fe(OH).sub.3 by the treatment
of aqueous 0.1-0.2M solution of Fe(III) salt, to advantage FeCl.sub.3,
with less than an equimolar amount of NH.sub.4OH, at 21.degree. C., under
sonication, to which a solution of a Fe(II) salt, to advantage
FeCl.sub.2, is added in the mole ratio Fe(III)/Fe(II)=2 under sonication
and the mixture is poured into five- to tenfold, to advantage eightfold,
molar excess of 0.5M NH.sub.4OH. The mixture is left aging for 0-30 min,
to advantage 15 min, and then the precipitate is repeatedly, to advantage
7-10 times, magnetically separated and washed with deionized water. Then
1-3 fold amount, to advantage 1.5 fold amount, relative to the amount of
magnetite, of 0.1 M aqueous solution of sodium citrate is added and then,
dropwise, 1-3 fold amount, to advantage 1.5 fold amount, relative to the
amount of magnetite, of 0.7 M aqueous solution of sodium hypochlorite.
The precipitate is repeatedly, to advantage 7-10 times, washed with
deionized water under the formation of colloidal maghemite to which,
after dilution, is added dropwise, to advantage under 5-min sonication,
an aqueous solution of a modification agent, in the weight ratio
modification agent/iron oxide=0.1-10, to advantage 0.2 for amino acids
and poly(amino acid)s and 5 for saccharides.
The particles smaller than 2 nm with polydispersity index smaller than 1.1
are prepared by mixing at 21.degree. C. 1 volume part of 10-60 wt. %, to
advantage 50 wt. %, of an aqueous solution of a saccharide, disaccharide
or polysaccharide, such as D-arabinose, D-glucose, D-galactose,
D-mannose, lactose, maltose, dextran and dextrins, and 1 volume part of
aqueous solution of a Fe(II) and Fe(III) salt, to advantage FeCl.sub.2
and FeCl.sub.3, where the molar ratio Fe(III)/Fe(II)=2. A 5-15%, to
advantage 7.5%, solution of NH.sub.4OH is added until pH 12 is attained
and the mixture is heated at 60.degree. C. for 15 min. The mixture is
then sonicated at 350 W for 5 min and then washed for 24 h by dialysis in
water using a membrane with molecular weight cut-off 14,000 until pH 7 is
reached. The volume of solution is reduced by evaporation so that the
final dry matter content is 50-100 mg/ml, to advantage 80 mg per 1 ml.
Superparamagnetic nanoparticle probes according to the invention can be
used for labelling cells used in magnetic resonance imaging for
monitoring their movement, localization, survival and differentiation
especially in detection of pathologies with cell dysfunction and of
tissue regeneration and also for labelling and monitoring cells
administered for cell therapy purposes, in particular embryonal stem
cells, fetal stem cells, stem cells of an adult human including bone
marrow stem cells, olfactory glial cells, fat tissue cells, in the
recipient organism by magnetic resonance.
The preparation of labelled cells proceeds by adding to the complete
culture medium 5-20 .mu.l, to advantage 10 .mu.l, of a colloid containing
0.05-45 mg iron oxide per ml, to advantage 1-5 mg iron oxide per ml of
the medium, and culturing the cells for a period of 1-7 days, to
advantage for 1-3 days, at 37.degree. C. and 5% of CO.sub.2.