Comparison of two main fragmentation methods of amyloid beta fibrils for establishing an Alzheimer disease model in cell culture
Acta Medica, vol.56, no.2, pp.92-107, 2025 (TRDizin)
- Publication Type: Article / Article
- Volume: 56 Issue: 2
- Publication Date: 2025
- Journal Name: Acta Medica
- Journal Indexes: TR DİZİN (ULAKBİM)
- Page Numbers: pp.92-107
- Hacettepe University Affiliated: Yes
Abstract
Aim: Amyloid beta
fibrils have been
shown to play
a role in
plaque formation and
aggregation in Alzheimer’s
disease. Obtaining these
fibrils using two
main methods and
applying them to
Alzheimer’s modelling is
crucial in understanding
the pathology of
the disease at
the molecular level
and identify in
therapeutic targets. The
aim of our
study is to determine the optimum sonication parameters using probe and
ultrasonic bath sonication laboratory methods and to demonstrate Alzheimer’s
disease modelling at the cellular level. Methods: Lyophilized
human peptide amyloid
beta1-42 fibrils (fAβ1-42)
were subjected to
probe sonication for
1 minute with
1, 3 and
5 second on/off
pulse applications at
varying ambient temperatures
(room temperature, ice
and ice surrounded
by dry ice
[ISDI]) for 20,
40 and 60
cycles, respectively. Then,
ultrasonic bath sonication
was performed in
10 °C water
for 1 hour.
The length of
the fragmented fibrils
was quantified by
transmission electron microscopy
(TEM). fAβ1-42 at
different concentrations was
applied to SH-SY5Y
cell line. The
non-toxic dose and time of fAβ1-42 application were analysed using the
WST-1 assay. Intracellular
and extracellular fibrils
were visualized with
immunofluorescence (IF) labelling. Results: Although,
fragmentation was observed
under all conditions,
it was observed
that fibrillar lengths
decreased as the
on/off pulse times
increased, regardless of
the number of
cycles with ice
and dry ice.
Additionally, decreasing the
temperature increased fibrillar
fragmentation. Conclusion: We anticipate that our study will contribute
to the literature by developing an
effective and economical
sonication method for
fibrillar fragmentation with two main laboratory methods and obtaining
fAβ1-42 that can be used in cells at optimum concentration.