The impact of storage duration of fresh human amniotic membrane with the level of EGF, TGF, and BFGF growth factors

Background: Human amnion membrane has been used in many regenerative medicine. It has been reported that fibroblast and other epithelial cells lost its viability after going through cryopreservation. The objective of this study is to prove the effect of storage duration of the fresh amnion membrane to the levels of EGF, TGF-b and bFGF growth factors. Methods: Eighteen fresh human amnion samples stored at Dr. Soetomo General Academic Hospital’s Centre of Biomaterial and Tissue Bank was involved in this study. Each group of 6 is selected from different storage times at -20° C (1 week, 3 months, and 6 months). The levels of EGF, TGF-b and bFGF growth factors at different storage times were analysed using ELISA. Result: From ELISA analysis, EGF mean value at 1 week storage duration was 8.43±5.13 pg/mL, 3 months storage was at 16.59±22.20 pg/mL and 9 month storage was 4.52±2.04 pg/mL. From TGF-b analysis it was 140.41±25.77 pg/mL at 1 week storage time, 3 months storage time was 140.58±50.62 pg/mL, and at 9 months storage it was 115.75±35.91 pg/mL. From the EGF and TGF-b analysis, it shows that the difference of the growth factor levels are negligible between the 3 storage duration groups, (p= 0.462) and (p=0.331) respectively. Whereas in bFGF , the mean value at 1 week storage duration was 57.60± 26.90 pg/mL, 3 months was 37.94±13.11 pg/mL, and at 9 months storage time it was 147.66±89.93 pg/mL. In comparison to 1 week group against 9 months, and 3 months against 9 months, results showed significant changes at p=0.028 and p=0.010 respectively (p<0,05). Whereas at 1 week against 3 months storage duration, the result was insignificant at p= 0.273 (p>0,05). Conclusion: There were no significant changes in the value of growth factor between different storage times at 1 week, 3 months, and 9 months.