[Investigation on the effect of trehalose on alpha-actinin in cryopreserved human skin].
OBJECTIVE
To compare the effect of trehalose with that of different traditional cryoprotectants on human skin and to detect the new protection mechanism of trehalose in hypothermia.


METHODS
The skins to be cryopreserved were first treated with DMSO/Propyleneglycol (D/P group), trehalose/DMSO (T/D group), DMSO/serumfree keratinocyte medium (D/K group), DMEM (DMEM group), respectively, so as to be compared with fresh skin (control grouop). Then the histological structure of skin of different groups was observed and analyzed by pathological technology (SP immunohistochemistry, DAB staining). Furthermore, the influence of trehalose on alpha-actinin at gene level with RT-PCR was investigated. The viability of skin in 5 respective groups was evaluated by using succinate dehydrogenase (SDH). The experiments were carried out 14 days after cryopreservation.


RESULTS
The results of immunohistochemistry showed that A values of control group, T/D group, D/P group, D/K group and DMEM group were 27.50 +/- 7.92, 18.40 +/- 5.81, 13.10 +/- 5.11, 11.50 +/- 4.54 and 5.30 +/- 2.14, respectively. There was no significant difference between control group and T/D group (P > 0.05), but control group was significantly different from the other groups (P < 0.05). The results of PCR studies showed that A values of control group, T/D group, D/P group, D/K group and DMEM group were 0.816 +/- 0.134, 0.723 +/- 0.245, 0.564 +/- 0.265, 0.245 +/- 0.071 and 0.148 +/- 0.048, respectively. Control group was not significantly different from T/D group and D/P group (P > 0.05), but was significantly different from D/K group and DMEM group (P < 0.05). The results of SDH showed that A valuse of control group, T/D group, D/P group, D/K group and DMEM group were 18.2 +/- 3.7, 12.3 +/- 3.6, 10.2 +/- 2.4, 7.3 +/- 2.1 and 5.7 +/- 1.5, respectively. There was no significant difference between control group and T/D group (P > 0.05), while control group was significantly different from the other groups (P < 0.05).


CONCLUSIONS
The results suggest that cryopreservation protocol-trehalose/DMSO is better than the traditional cryoprotectant for cryopreservation on alpha-actinin of human skin.
Burns Institute, First Affiliated Hospital to PLA General Hospital, Beijing, 100037, P. R. China. jxm304@126.com
Xiaoming
X
Jia
Yu
Y
Yu
Dongjie
D
Li
D000185:Actinin / Q000096:biosynthesis*; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000494:pharmacology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D006801:Humans; D007150:Immunohistochemistry; D019946:Propylene Glycol / Q000494:pharmacology; D012867:Skin / Q000187:drug effects / Q000378:metabolism*; D014021:Tissue Preservation / Q000379:methods*; D014199:Trehalose / Q000008:administration & dosage / Q000494:pharmacology*
D003451:Cryoprotective Agents; D000185:Actinin; D019946:Propylene Glycol; D014199:Trehalose; D004121:Dimethyl Sulfoxide