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纳米珍珠粉对大鼠深Ⅱ°烧伤创面愈合的影响
作者：崔淑芳，郝光荣，汤 球，孙 伟，曹 平
【摘要】 目的 研究纳米珍珠粉对大鼠深Ⅱ°烧伤创面愈合的影响并与微米珍珠粉在烧伤愈合中进行效比较。方法 纳米珍珠粉、微米珍珠粉(0.5g/只)分别与甘油按1∶1配制成混合液。以微米珍珠粉组与自然愈合组作对照，纳米珍珠粉组为实验组。测定大鼠14%深Ⅱ°烫伤后7、15、21天的创面愈合率、创面组织学观察以及皮肤羟脯氨酸的含量。结果 纳米珍珠粉组创面愈合率较微米珍珠粉组、自然愈合组高(P＜0.05或P＜0.01)；烧伤后3天各组皮肤羟脯氨酸含量均低于正常(P＜0.01)，纳米珍珠粉组减少程度小于其他组；伤后5天各组HP含量有所恢复，尤以纳米珍珠粉组显著高于自然愈合组及微米珍珠粉组；10天后各组明显升高，而纳米珍珠粉组HP含量的升高差异有非常显著性(P＜0.01)。结论 纳米珍珠粉可以促进大鼠深Ⅱ°烧伤创面的愈合，其愈合作用明显高于微米珍珠粉(P＜0.05)。
【关键词】 纳米珍珠粉；微米珍珠粉；烧伤；羟脯氨酸
【Abstract】 Objective To study the efficiency comparison between nano and micro pearl powder for treatment of deep Ⅱ° skin scalds in rats.Methods Nano and micro pearl powder (0.5g each rat) are mixed with glycerol (1∶1).With the micro pearl powder group and the normal healing group as the control，nano pearl powder were used as drug on the deep Ⅱ° skin scalds of rats in the experimental groups.Wound healing rates was measured at 7，15，21 days post-injury；and the content of hydroxyproline (HP) were detected also.Results Wound healing rates of the nano pearl powder group were higher than that of the micro pearl powder group and the normal healing group (P＜0.05 or P＜0.01).At the 3th day post-injury，the content of HP in every group were lower than that in the normal group (P＜0.01)，while the decrease in the nano pearl powder group was slight than those in other groups.At the 5th day post-injury，the content of OHP recovered to some extent，with a significantly higher elevation in the nano pearl powder group than those in other groups.At the 10th day post-injury，evident HP increase was found in every group，with a statistical significant higher elevation in the nano pearl powder group (P＜0.01).Conclusion Nano pearl powder could promote the wound healing of deep Ⅱ° skin scalds in rats significantly，it’s healing was more higher than that of micron pearl powder.
【Key words】 nano pearl powder；micro pearl powder；scald；hydroxyproline(HP)
This research is to study the wound healing efficacy of nano pearl powder in treatment of deep Ⅱ°skin scalds in rats and compare the function with those of micron pearl powder.
1 Material and methods
1.1 Pearl powder (Zhejiang Fenix Pharmaceutical Co.，Ltd)，nano and micro pearl powder (0.5g each rat ) are mixed with glycerol (1∶1).Dermal hydroxyproline detection kit (Nanjing Jiancheng biotechnology Institute，Approved Number 20040631).
1.2 Experimental animals Female SD rats (n＝190，body weight 240～260g，Experimental Animal Center，Second Military Medical University).The rats were fedin metabolic cages for 1 week.
Qualified certificate for experimental animal ［SCXK(Hu)2002-0006］；qualified certificate for animal facility ［No.SCXK(Hu)2002-0026］.
1.3 Establishment of deep Ⅱ° skin scalds model in rats The model was made based on the traditional method set up by Yang Taimei［1］.The rats were denuded with BaS and scalded for 15s in 80℃ water the next day to get deep Ⅱ° skin scalds model in rats with a wound accounting for 14% of the body surface.See fig.1.
1.4 Methods Those rats were divided into 3 groups after the burn (n=60)：nano pearl powder group，micro pearl powder and normal healing group.In nano pearl powder and micro pearl powder groups the wound were daubed with a mixture of nano，micro pearl powder and glycerol once a day，while the rats in the normal healing group were fed regularly.The experiment lasted 21 days.
1.5 Observations
1.5.1 Wound healing circumstance The wound development healing percentage and healing circumstance were observed in the whole healing.The healing rates were calculated on day 7、15、21 after scalding.Wound healing rate=healing surface/original skin lesion×100%.Regarding the wound healing as the index，to judge the healing time.
1.5.2 Dermal hydroxyproline(HP)measurement Tissue in the wound were taken from the 3 groups of rats on day 3，5，10，15 and 21 after scalding with normal rats skin as a control.The dorsal skin were denuded and separated from the connective tissue followed by a drying at 80℃，50mg of which were measured by dermal HP reagent kit.HP content was recorded by μg/mg.
1.6 Data statistics and analysis Inputting the data into the computer，to analyze the data with the Excel software，a significance dissimilarity of group data was showed P＜0.05 or P＜0.01，the average data was showed x±s.
2 Results
2.1 Comparison of the healing time and the wound development healing rate The skin wound in the nano pearl powder group is clear with no effusion and infection.In the micro pearl powder group，there is a rat with effusion on the skin surface.In the control group 2 cases of effusion and one infection were found.
The wound healing time of nano pearl powder has a significance dissimilarity more than that of micro pearl powder (P<0.01)，the wound development healing rate is more quickly.(P<0.01，See table 1 for details).
Tab.1 Comparison of wound healing time and development healing rate of the different times (x±s，%)
0Note：compared with control group：*P＜0.05，**P＜0.01；compared with micro pearl powder group：△P＜0.05，△△P＜0.01
2.2 Changement of the scalded skin tissue HP content in different phases The HP content in the normal control group is (78.05±7.79)μg/mg.On day 3 after the scald，the HP content in every group of animals were lower than that in the normal group (P＜0.01)，while the decrease in the nano pearl powder group was slight than those in other groups.On day 5，the HP recovered to some extent，with a significantly higher elevation in the nano pearl powder group than those in other groups.On day 10，evident HP increase was found in every group，with a statistical significant higher elevation in the nano pearl powder group (P<0.01).See table 2.
Tab.2 Changement of the scalded skin tissue HP content in different phases (x±s，n=10)
Normal control value：78.05±7.79；t test：compared with normal control：△P＜0.05，△△P＜0.01；compared with experimental control*P＜0.05，**P＜0.01；compared with micro pearl powder：☆P＜0.05，☆☆P＜0.01
3 Discussion
By the measurement of the wound healing rate，nano pearl powder had the function of promoting obviously the wound healing in earlier period，it’s curative effect is obvious better than that of micron pearl powder by comparison (P＜0.05).The scald healing is a compound process of inflammatory reaction，cell proliferation，re-epithelization and collagen metabolizing［2］.The collagen synthesized and secreted by fibroblasts is the main structure protein of the skin，and HP is a steady component of collagen，which accounts for 13.4% of the collagen proteins.The healing of a burn lesion mainly relies on the collagen fiber formed by the granulation tissue，thus，measurement of the variance of skin HP content may act as a specific and sensitive marker for wound healing in tested animals［3］.
The decrease of HP in the early stage of scald has much to do with the collagen decomposing caused by heat damage and the eschar formed by collagen degeneration［4］.The dermal HP content in the nano peal powder group started to increase on day 5，reached the same value as the normal control group on day 10，and arrived a peak value on day 15.Between day 3 and 21，the dermal HP content in the nano pearl powder group is significantly higher than those in micro pearl powder group and normal healing group，which indicates that nano peal powder has a more potent stimulation effect on collagen synthesis and a higher absorption rate than micro pear powder.So，nano peal powder can promote the healing process of a scalded tissue by relieving the decrease of HP in the affected skin and accelerating the synthesis of collagen fiber.
Fig1a''1b The trauma of deep Ⅱ°degree wound in SD rats(×100)
【参考文献】
1 YANG Tai-mei，FENG Shi-jie，XU Sa-ying.Making the rat scald model.Acta Universitatis Medicinalis Secondae Shanghai，1997，173)：91.
2 Portera CA，Love EJ，Memore L，et al.Effect of macrophagge stimulation on collagen biosynthesis in the healing wound.Am Surg，1997，63(2)：125-131.
3 Edwards CA，Orien JR.Modified assay for determination of hydroxyproline in tissue hydrolyzate.Clin Chem Acta，1980，104：161.
4 Nussbaum MS.Arginine stimulates wound healing and immune function in elderly humen beings.J PEN，1994，18：194.
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