几丁聚糖纳米微粒分散体系应用于烧伤创面的实验

Application of chitosan nanoparticles dispersion system in skin burn

目的:制备以几丁聚糖为主要原料纳米微粒分散体系,并通过动物烧伤实验进行医学应用效果观察。方法:实验于2003-05/2004-05在浙江省医学科学院医学生物工程研究所及浙江省动物实验中心完成。首先运用物理和化学综合手段,制备几丁聚糖纳米微粒分散体系,用透射电镜观察法进行纳米微粒尺寸测定评估;其次采用制备的几丁聚糖纳米微粒分散体系进行动物烧伤实验,①烧伤动物模型建立:选择体质量2.5～3.0kg健康兔子8只,雌雄对半,脱毛并麻醉后,在实验兔子背部,以脊椎为对称轴,用盛装100℃开水的人工烧伤器(质量500g),紧贴兔子皮肤3s,制成浅Ⅱ度烧伤创面24个,5s制成深Ⅱ度创面24个,每个创面平均面积16cm2,左侧创面为实验组,右侧为对照组。②治疗方法:左侧实验组创面涂敷几丁聚糖纳米体系,右侧创面涂敷凡士林,2次/d,上下午各1次,连续3d后停止涂敷,每日观测记录创面的渗出液、创面大小、结痂时间、脱痂愈合时间等临床指标。结果:①几丁聚糖纳米微粒分散体系颗粒粒径平均为(36.40±5.14)nm。②治疗动物烧伤实验第7天的烧伤创面平均面积试验组小于对照组[浅Ⅱ度:(2.29±0.51)和(4.28±0.45)cm2;深Ⅱ度:(2.15±0.34)和(4.39±0.52)cm2,P<0.01]。③烧伤创面结痂平均时间试验组短于对照组[浅Ⅱ度:(6.45±0.58)和(10.5±0.75)d;深Ⅱ度:(7.5±0.66)和(10.6±0.78)d,P<0.01]。④几丁聚糖纳米体系对烧伤创面脱痂愈合的平均时间试验组短于对照组[浅Ⅱ度:(14.36±0.58)和(16.27±0.75)d;深Ⅱ度:(22.0±1.33)和(29.88±1.86)d,P<0.01]。结论:几丁聚糖纳米微粒分散体系对动物Ⅱ度烧伤具有明显的缩小创面、提早结痂、促进愈合之疗效,提示具有临床功能敷料的开发应用前景。

AIM： To produce a nanoparticles dispersion system with chitosan as the main component, and observe the curative effect on animal skin burn. METHODS： The experiment was carried out in the Institute of Medical Bioengineering, Zhejiang Academy of Medical Science and Zhejiang Animal Experimental Center between May 2003 and May 2004. At first, chitosan nanoparticle system was produced by physical and chemical methods, the diameter of nanoparticle was measured by transmission electron microscopy; Secondly, the clinical curative effect was evaluated through curing animal skin burn experiment. ① Establishment of animal model of burn： Eight （4 males and 4 females） healthy rabbits of 2.5-3.0 kg） were selected. Furs of rabbits back were shaved completely. After anesthesia, a special tool of burn that contained boiling water （100℃, weighing 500 g） tight contact the back skin of rabbits for 3 seconds （to make superficial second degree burn, n=24） and 5 seconds （to make deep second degree burn, n=24）. The average area of wound was 16 cm^2. The left side was taken as the experimental group and the right side as the control group. ② Therapeutic methods： The wounds of left side on back skin were dressed with chitosan nanoparticles system and those of right side were dressed with vaseline, twice a day in the morning and afternoon respectively for 3 continuous days, and the clinical indexes of wound exudates, size of wound, scabbing time and time for decrustation and healing were observed and measured every day. RESULTS： ① The chitosan nanoparticles＇ average diameter was （36.40 ±5.143） nm. ② The average areas of burn at 7 days were smaller in the experimental group than in the control group [superficial second degree burn： （2.29±0.51）, （4.28±0.45） cm^2; deep second degree burn： （2.15±0.34）, （4.39±0.52） cm^2, P 〈 0.01]. ③ The average scabbing times of wound were shorter in the experimental group than in the control group [superficial second degree burn： （6.45±0.58）, （10.5±0.75） days; deep second degree burn： （7.5±0.66）, （10.6±0.78） days, P 〈 0.01]. ④ The average time for decrustation and healings of wound were shorter in the experimental group than in the control group [superficial second degree burn： （14.36±0.58）, （16.27±0.75） days; deep second degree burn： （22.0±1.33）, （29.88±1.86） days, P 〈 0.01]. CONCLUSION： The chitosan nanoparticles dispersion system has evident curative effects for minifying the wound area, the scabbing time and the time for decrustation and healing, which suggests that the system can be developed a new medical prospect.