THE EFFECT OF CHITOSAN ON OSTEOCLAST VIABILITY, BONE RESORPTION AND RADICAL OXYGEN PRODUCTION OF PRIMARY OSTEOCLAST CULTURE OF MOUSE BONE
Abstract
Objective: The purpose of this investigation was to determine the effect of chitosan on osteoclast cells by observing cell viability, bone resorption,
and radical oxygen species (ROS) production.
Methods: Osteoclast cells were obtained from the primary culture of bone marrow mouse. The osteoclast cells were identified by tartrate-resistant
acid phosphatase (TRAP) marker both on the cells and the culture medium. The osteoclast cell viability was observed with (3-(4,5-dimethylthiazol2-yl)-2,5-diphenyltetrazolium
bromide)
tetrazolium
assay
and Bradford
assay
for
total
protein
medium
culture,
while
ROS
production
was
measured
with
malondialdehyde
(MDA)
assay.
Slices
of cow
cortical
bone
were
used as a substrate
for
osteoclastic
resorption
and concentrated
hydrochloric
acid
were
used to
activate
resorption
and pit formation
by
any
osteoclasts.
Results: Osteoclast cells were identified by TRAP marker and chitosan treated group cells showed lower optical density value compared to control
(p<0.05) on TRAP assay medium culture. Cell viability indicated lower on chitosan group than control (p<0.05). There was a qualitative difference of
the pits formed on the bone surface between the control and the chitosan group. There was a significant difference in MDA (mmol/ml) between the
control and the chitosan group with (p<0.05).
Conclusion: Based on this research, we conclude that chitosan inhibits the viability of osteoclast cells, decreases ROS production and bone resorption.
Keywords: Chitosan, Osteoclast proliferation, Radical oxygen species, Bone resorption.
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