interTALENT UNIDEB 2017

interTALENT UNIDEB 2017. február 23-24. Debreceni Egyetem
First sterile implans are incubated with Candida
albicans SC5314 for 24 hours solely in 24 well
plates. Then implants are washed repeatedly with
saline. After drying samples are incubated with
crystal violet solution for 15 minutes. Unbound
crystal violet is washed out with saline. Crystal
violet bounded by the biofi lm is dissolved with 95
v/v% ethanol and 100 μl is taken into 96 well plate.
Absorbance is measured.
MTT CELL VIABILITY TEST
MTT assay is a colorimetric cell viability test.
Only viable cells are able to convert the
originally yellow colored and water soluble
MTT dye (3-(4,5-dimethylthiazol-2-yl))-2,5-
diphenyltetrazolium bromide) to a purple and
insoluble formazan salt by their mitochondrial
dehydrogenase enzymes. The formazan salt can
be dissolved in the mixture of hydrochloric acid
and isopropyl alcohol than absorbance can be
measured.
METHOD
CaCo-2 cell culture is used as the model of human
intestinal endothelium. Cells were seeded in a 96
well plate (1*10^3/well) until monolayer formation.
The samples are sterilized with 70% ethanol and
UV-light.
Then samples are placed to sterile tubes and
submerged in 2 ml of DMEM medium. The control
is 2 ml of medium with no added material. Samples
are incubated on 37 °C in 5% CO2. Cell viability
tests are performed on the 4., 8. and 12. days, 1., 2.
and 3. month.
First medium is added to the cells and incubated
for 3 hours on 37 °C. Then MTT dye is applied
and incubated for 30 minutes on 37 °C. Finally,
absorbance is measured which is in correlation with
the cell viability.
2. Figure: Cell viability % in the 4. day with 13
samples and the control
SUMMARY
Biocompatibility examination is inevitable through
the development of implants because implants
are inserted into the human body so it must be
biologically compatible with the tissues.
Measured results in the MTT cell viability test
indicates us that most of the implants do not
infl uence the CaCo-2 cell viability.
Biofi lm formation results enable us to restrict
the number of applicable implants to further
examinations.
We intend to make examinations on diff erent cell
cultures in MTT assay, in vivo animal tests and
histological research.
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