[Comparative study of the effect of different cryoprotectants on desmoglein 1 in human skin].
OBJECTIVE
To compare the effects of two different cryoprotectants on human desmoglein 1 (Dsg 1), and to provide experimental basis for the optimization of cryoprotectant.


METHODS
Five donated thin split-thickness skin grafts were used, and the experiment was conducted within 4 hours after skin grafts harvest. The skin grafts were divided into 3 groups: group A (n = 2) in which skin grafts were immersed in 0.5 mol/L trehalose/DMSO; group B (n = 2) in which skin grafts were immersed in DMSO/propanediol; group C (n = 1) in which fresh skin graft received no further treatment. Groups A and B were stored in -196 degree C liquid nitrogen for 7 and 21 days, respectively, and then underwent experiment. Immunohistochemistry staining observation was performed on each group, RT-PCR method was used to detect the expression of Dsg 1 in skin.


RESULTS
The immunohistochemistry staining showed that the protein in groups A and B was stained brown-yellow and distributed evenly 7 days after cryopreservation; the expression signal of epidermal basal cell was similar to that of group C; absorbance (A) value of groups A, B and C was 0.285 +/- 0.006, 0.284 +/- 0.004 and 0.287 +/- 0.008, respectively, suggesting there was no significant difference between groups A and B and group C (P > 0.05). At 21 days after cryopreservation, the expression of positive cells in group B decreased; no obvious decrease was observed in group A, A value of groups A and B was 0.282 +/- 0.004 and 0.275 +/- 0.005, respectively, indicating there was a significant difference between group B and groups A and C (P < 0.05). RT-PCR detection showed that A value of groups A and B at 7 days after cryopreservation was 0.810 +/- 0.012 and 0.803 +/- 0.008, respectively; A value of groups A and B at 21 days after cryopreservation was 0.806 +/- 0.008 and 0.782 +/- 0.013, respectively; and the A value of group C was 0.814 +/- 0.012, indicating there was significant difference between group B and groups A and C at 21 days after cryopreservation (p < 0.05), and no significant differences among groups were noted at other time points (P > 0.05).


CONCLUSIONS
Trehalose/DMSO is better than traditional cryoprotectant DMSO/propanediol in protecting Dsg 1 of human skin.
Jia
Xiaoming
X
Burns Institute, the First Affiliated Hospital, General Hospital of PLA, Beijing, 100048, PR China. jxm304@126.com
Li
Dongjie
D
Ma
Caihong
C
D003451:Cryoprotective Agents / Q000494:pharmacology*; D051183:Desmoglein 1 / Q000096:biosynthesis*; D006085:Graft Survival; D006801:Humans; D012867:Skin / Q000187:drug effects* / Q000378:metabolism*; D013524:Surgical Flaps; D046509:Tissue Culture Techniques; D014199:Trehalose / Q000494:pharmacology
D003451:Cryoprotective Agents; D051183:Desmoglein 1; D014199:Trehalose