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The Effects on Collagen and Actin of Expanded Skin in Different Expanded Ways

Author: ShiWenBin
Tutor: DongYuYing;DongFuSheng
School: Hebei Medical University
Course: Clinical Stomatology
Keywords: Skin expansion Collagen actin Immumohistochemistry Picrosirius-red
CLC: R622
Type: Master's thesis
Year: 2007
Downloads: 56
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Abstract


Objective: The sirius-red specific staining was applied to observe the change of collagen and its proportionality. Immunochemistry method was used to detect the expression of actin in expanded rabbit skin. This study aimed to understand the effects on collagen and actin of expansion skin in repeatedly rapid expansion method, rapid expansion method and conventional expansion method and to provide an experiment foundation for the suitable expanded method clinically.Methods: A total of 62 male New Zealand white rabbits were collected in this study. All animals were divided into five groups randomly: repeatedly rapid expansion (groupA), rapid expansion (groupB), conventional expansion (groupC), embedding un-expansion (groupD) and normal control (groupE). The rabbits in group A, B, C, D were anesthetized after weighed. Fifty millilitre expanders were embeded between the head skin and cranial bones. Anti-infective therapy was performed after operation.Since the third days after operations, expansion was begun in group A, B, C. Group A: Twenty millilitres physiological saline was injected into expander and maintained for 30 min and then was pumdbacked compeletly. There was a three hours’ interval to the next affusion.This expansion model was repeated for three times. But there was 15ml physiological saline to be pumpbacked last time and 5ml were reserved in the expander. The same expansion process was repeated for 10 days. Fifty millilitre of physiological saline was reserved in the expander at the maintaining period.Group B: Five millilitre of physiological saline was injected into the expander once a day, but the saline was not pumpbacked. The same expansion process was repeated for 10 days and 50ml physiological saline was reserved in the expander at the maintaining period. Group C: Five millilitre of physiological saline was injected into the expander every 3 day, but the saline was not pumpbacked. The same expansion process was repeated for 28 days and 50ml physiological saline was reserved in the expander at the maintaining period. Group D: The expanders were embeded but not affused. Certain of rabbits were selected randomly at the different maintaining period and the specimens were collected from the expanded kin. All specimens were fixed in formaldehyde fluid, embedded by paraffin and 4-μm-thicks sections were cut. The sirius-red specific staining were applied to observe the changes of type I collagen and type III collagen and their proportion while immunochemistry method was used to detect the expression of actin in expanded rabbit skin.All data are presented as Mean±SD. Statistical analysis was performed with SPSS 11.5 software, using ANOVA. Values for P<0.05 were considered significant difference. Results:1 Picrosirius-red specific staining of collagenThere were no significant difference between D and E at the different maintaining periods. Type I collagen was the main content and arrange regularly while type III collagen is the minor part in demal. Type I collagen arranged turbulently and was thicker and type III collagen decreased relatively at earlier maintaining period. After the maintaining period of 4 week, type I collagen arranged regularly and type III collagen was relatively increased.2 Semiquantitative analysis of collagen by specific staining of picrosirius-redComparisons of the area about type I collagen:The area of type I collagen in groupA, B, C were higher than that of group D respectively at the different maintaining period. There were significant difference between groupA,B,C and group D (P<0.05). At the same maintaining period, the area of type I collagen in group C are maximum , secondly A , thirdly B, but there were no significant difference among the group A,B and C(P>0.05). With the extension of maintaining period, the area of type I collagen increased gradually, but there was inclined to stabilize at the fourth week in group A, B. There were no significant difference between the fourth-week and the sixth-week maintaining period (P>0.05). Significant difference was observed among other maintaining period. With the extension of maintaining period, the area of type I collagen was increased gradually, but there was inclined to stabilize about the third week in group C. There were no significant difference between the third-week and the fourth-week maintaining period (P>0.05),but significant difference was observed among other maintaining period.Comparisons of the area about type III collagen:The area of type III collagen in group A, B, C was higher than that of group D respectively. There were no significant difference between the one-week maintaining period and control group(P>0.05), but other periods were significant difference (P<0.05). At the same maintaining period, the comparison of the area about type III collagen in group D is maximum,secondly A,thirdly B.There were no significant difference between group A, B, C at the same maintaining period (P>0.05).With the extension of maintaining period, the area of type III collagen was increased gradually, but there was inclined to stabilize at the fourth week in group A,B.There were no significant difference between the four-week and the six-week maintaining period (P>0.05),but significant difference were observed among other maintaining period. With the extension of maintaining period, the area of type III collagen increased gradually, but there was inclined to stabilize at the third week in group C. There were no significant difference between the third-week and the fourth-week maintaining period (P>0.05) in group C, but significant difference was observed among other maintaining period. Comparison of the proportion of collagen I and collagen III: The proportion of collagen I and collagen III in groupA, B, C are higher than that of group D respectively at the different period. There were significant difference between group A,B and group D at the first-week, second-week, third-week maintaining period (P<0.05), but no significant difference was observed after the fourth week(P>0.05). There were significant difference between group C and control group at the first-week,second-week maintaining period (P<0.05), but no significant difference were observed after three week (P>0.05).3 Immunohistochemistry results of actin:The positive expression rates of actin in group A, B, C were higher than that of group D respectively. There were significant difference between group A, B, C and group D(P<0.05). At the same maintaining period, significant difference were observed among group A, B, C(P<0.05). The positive expression rates of actin in group C were lower than that of group A at the same maintaining period.There were significant difference at the first-week, second-week, third-week maintaining period (P<0.05),but no significant difference was observed after the fourth-week maintaining period (P > 0.05).The positive expression rates of actin in group A were lower than that of group B at the same maintaining period. There were significant difference at the same maintaining period(P<0.05). With the extension of maintaining period, the positive expression rates of actin decreased gradually in groupA, B, C.Conclusion1 Different expanded ways have effects on the type I, III collagen and actin in the expanded skin.2 Quality of expanded skin in repeatly rapid expansion is better than rapid expansion at the same maintaining period.3 The repeatly rapid expansion may shorten the clinical period and acqure the same skin as the conventional expansion.4 The repeated rapid expansion can not only shorten the clinical period but also warrant the quality of skin flap. It is a safe and feasible method clinically.

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