Biomedical Engineering Reference
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On TCPS, the number of adherent cells was clearly greater than
that on PMB30. The cells that adhered on the PMBs were round in
shape, but thoseon TCPS had a spreading morphology.
The expression of IL-1
β
mRNA in the cells cultured on the
MPC polymers was then evaluated using reverse transcriptase-
polymerase chain reaction (RT-PCR) analysis.
29
HL-60 cells were
selected because they can differentiate into macrophage-like cells.
Inaddition,weassessedthecelladhesionandproteinadsorptionin
order to consider the relationship to the expression of IL-1
β
mRNA.
The procedure was based on that reported by Kishida
et al
.
24
,
25
The products of the PCR reaction were confirmed to be those
of the gene transcripts by detection of an 821-bp band (
β
-actin
mRNA) and a 566-bp band (IL-1
β
mRNA). The expression of the IL-
1
β
mRNA was determined as the relative value to that obtained for
β
-actin. In general, amplified
β
-actin mRNA was used as an internal
standard for the semi-quantification.
30
,
31
RT-PCR analysis is a highly sensitive method and can detect var-
ious mRNAs from a small number of cells in contact with polymeric
materials. As shown in Fig. 19.6, adherent HL-60 cells on the PMBs
showed a lower expression of IL-1
β
mRNA than did those on other
reference polymers. Kishida
et al
. reported the expression of IL-1
β
mRNAfromHL-60cellsthatadheredonvariouspolymersurfaces.
32
β
Figure 19.6.
ExpressionofIL-1
mRNAtranscriptsinHL-60cellsonpoly-
β
mer surfaces as a standard of
-actin by RT-PCR after 24 h incubation.
Abbreviation
: N.D., not detected.
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