Mat Yatim, Rusidah
(2015)
The effect of gamma irradiation on selected growth factors and receptors mRNA in glycerol cryopreserved human amniotic membrane.
Masters thesis, Universiti Sains Malaysia.
Abstract
Human amniotic membrane (HAM), due to its high biocompatibility, low
immunogenicity, anti-microbial, anti-viral properties as well as the presence of its
growth factors, has been used in various clinical applications. These growth factors
are key factors in regulating many cellular processes such as cellular growth,
proliferation and cellular differentiation. The current study aimed to explore the
effect of glycerol cryopreservation and gamma irradiation on the selected growth
factors and receptors mRNA present in HAM. Eight growth factors, namely, EGF,
HGF, KGF, TGF-α, TGF-β1, TGF-β2, TGF-β3 and bFGF and two growth factor
receptors, HGFR and KGFR were evaluated in this study. The total RNA was
extracted and converted to complimentary DNA using commercial kits.
Subsequently, the mRNA expressions of these growth factors were evaluated using
quantitative PCR and the results were statistically analyzed using REST-MCS
software. This study indicated the presence of these growth factors and receptors
mRNA in fresh, glycerol cryopreserved and irradiated glycerol cryopreserved HAM.
In glycerol cryopreserved HAM, the mRNA expression showed up-regulation of
HGF and bFGF and down-regulation of the rest of 8 genes which were EGF, HGFR,
KGF, KGFR, TGF-α, TGF-β1, TGF-β2 and TGF-β3. Interestingly, the glycerol
cryopreserved HAM radiated with 15 kGy showed up-regulation in the mRNA
expression of 7 genes, namely, EGF, HGF, KGF, KGFR, TGF-β1, TGF-β2 and
TGF-β3 and down-regulated mRNA expression of HGFR, TGF-α and bFGF.
However, these mRNA expressions did not show a statistically significant difference
compared to control groups. Thus, it can be concluded that the glycerol
cryopreservation did not have an effect on the growth factors’ and receptors’ mRNA
expression levels in HAM. Similarly, 15 kGy gamma irradiation did not have an
effect on the growth factors’ and receptors’ mRNA expression in glycerol
cryopreserved HAM. This finding provides a useful information to clinicians and
surgeons to choose the best method for HAM preservation that could benefit patients
in their treatment.
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