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Biochemical Indicators of Implantation Success of Tissue-Engineered Oral Mucosa.
[severe combined immunodeficiency]
Real-time
(
RT
)
determination
of
the
health
of
in
vitro
tissue-engineered
constructs
prior
to
grafting
is
essential
for
prediction
of
success
of
the
implanted
tissue-engineered
graft
.
In
addition
,
the
US
Food
and
Drug
Administration
requires
specific
release
criteria
in
RT
prior
to
the
release
of
tissue-engineered
devices
for
human
use
.
In
principle
,
assessing
the
viability
and
functionality
of
the
cellular
component
can
be
achieved
by
quantifying
the
secretion
of
growth
factors
and
chemokines
of
tissue-engineered
constructs
.
Ex
vivo-produced
oral
mucosa
equivalents
(
EVPOMEs
)
were
fabricated
under
thermally
stressed
conditions
at
43
°
C
for
24
h
to
create
a
functionally
compromised
EVPOME
.
We
used
microchannel
enzyme-linked
immunosorbent
assay
to
evaluate
the
functionality
of
the
cellular
component
,
oral
keratinocytes
,
of
stressed
and
unstressed
EVPOMEs
by
measuring
the
release
of
vascular
endothelial
growth
factor
(
VEGF
)
,
interleukin-
8
(
IL
-
8
)
,
human
β-defensin
1
(
hBD-
1
)
,
and
tissue
inhibitor
of
metalloproteinase
1
and
2
(
TIMP-
1
and
-
2
)
into
the
spent
medium
,
which
was
collected
on
the
same
day
prior
to
graft
implantation
into
severe
combined
immunodeficiency
mice
.
Implanted
EVPOMEs
'
histology
on
the
seventh
postimplantation
day
was
used
to
correlate
outcomes
of
grafting
to
secreted
amounts
of
IL
-
8
,
hBD-
1
,
VEGF
,
TIMP-
1
,
and
TIMP-
2
from
corresponding
EVPOMEs
.
Our
findings
showed
that
significantly
higher
levels
of
IL
-
8
,
hBD-
1
,
and
TIMP-
2
were
secreted
from
controls
than
from
thermally
stressed
EVPOMEs
.
We
also
found
a
direct
correlation
between
secreted
VEGF
and
IL
-
8
and
blood
vessel
counts
of
implanted
EVPOMEs
.
We
concluded
that
measuring
the
constitutive
release
of
these
factors
can
be
used
as
noninvasive
predictors
of
healthy
tissue-engineered
EVPOMEs
in
RT
,
prior
to
their
implantation
.
Diseases
Validation
Diseases presenting
"blood vessel counts"
symptom
severe combined immunodeficiency
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