壳聚糖成膜剂联合EGF在兔深Ⅱ度烫伤创面愈合的实验研究

发布时间：2018-05-03 21:05

本文选题：壳聚糖 + 表皮生长因子　；参考：《泸州医学院》2013年硕士论文

【摘要】：目的：本实验通过离体细胞水平和建立兔深Ⅱ度烫伤自身对照模型，研究壳聚糖成膜喷剂联合表皮细胞生长因子喷雾剂（EGF）对兔深Ⅱ度烫伤创面的早中期的治疗效果及其创面愈合的相关影响机制。通过观察创面愈合的大体肉眼观、创面愈合率的计算、痂下细菌计数、创面病理学观察、创面组织羟脯氨酸(Hyp）含量变化、ki67在创面的分布和表达水平，探讨联合用药在烫伤早中期治疗的协同优势及其应用前景。方法：选取健康清洁级新西兰大白兔24只，以兔脊柱为中间线在背部两侧用水烫法建立6个对称的兔深Ⅱ度烫伤创面，上端两个创面为实验组（A组：壳聚糖成膜剂联合EGF组），中间两个创面为对照组1（B组：EGF组），下端两个创面为对照组2（C组：壳聚糖组），A组致伤后，先将EGF喷涂于创面，至药液不外流为止，1分钟后再将壳聚糖喷涂于创面，待其自然成膜后，，包扎固定。B组致伤后,仅在创面喷涂EGF至药液不外流为止，包扎固定。C组致伤后，仅在创面喷涂壳聚糖，待其自然成膜后，包扎固定。每24h创面再次给药，更换外敷料，肉眼观察三组创面大体形态变化及愈合情况。选取3d、7d、10d、14d四个观察时相点，每个时相点随机抽取6只兔子，将抽取的6只兔子空气栓塞处死，用称量纸描绘计重得出创面面积并计算创面愈合率。在严格无菌操作下，切取烫伤创面焦痂及痂下组织块1.0g，处理后行培养，计算每克创面组织的细菌群落总数。再取兔子背部创面100mg组织，处理后行组织羟脯氨酸（Hyp）含量测定。剩余的标本组织一部分切片HE染色后观测病理学指标，一部分标本行ki67特异性抗体染色后观察免疫组化指标。结果：创面大体观察，A组创面较B、C两组创面，创周炎性反应轻，痂壳松软，创面缩小速度快。创面愈合率的比较，A组明显高于B、C两组，B、C两组之间的差异则不明显。创面组织病理学标本镜下观察见，A组创面较B、C两组创面，坏死组织与创面附着松散，炎性细胞趋化聚集速度快，内皮细胞增生明显，胶原纤维增生活跃，新生上皮排列整齐。痂下细菌菌落数（CFU/g）阳性检出率A组阳性检出率低于B组，B组高于C组，A、B两组差异不明显。组织羟脯氨酸（Hyp）含量检测，A组Hyp含量增长速度快于B组，C组优于B组，A、C两组之间差异不大。ki67免疫组化分析，A组创面较B、C两组创面，阳性细胞聚集紧密，量多，着色深，排列整齐。ki67指数阳性率比较，A、B两组之间差异不明显，A、B两组与C组的差异均较明显。结论：两种药物联合使用能够加强兔深Ⅱ度烫伤创面的愈合能力，并能弥补单一用药的缺陷。能全面发挥其抑制创面细菌生长，加快上皮细胞增殖生长，促进胶原蛋白的合成的作用。
[Abstract]:Objective: to establish an autocontrol model of deep second degree scalding in rabbits at the level of in vitro cells. To study the effect of chitosan film forming spray combined with epidermal growth factor spray (EGF) on rabbit deep second degree scald wound in early and middle stage and the related mechanism of wound healing. By observing the gross naked view of wound healing, the calculation of wound healing rate, the count of subeschar bacteria, the pathological observation of wound, the content of hydroxyproline in wound tissue, the distribution and expression level of Yiki67 in the wound were observed. To explore the synergistic advantage and application prospect of combined medication in early and mid-term scalding. Methods: 24 healthy clean grade New Zealand white rabbits were used to establish six symmetrical deep second degree scald wounds with spinal cord as the middle line on both sides of the back. The upper two wounds were treated as experimental group A: chitosan film forming agent combined with EGF group, the middle two wounds were the control group 1 B: 1% EGF group, and the lower two wounds were the control group 2 C group: after the injury in chitosan group A group, EGF was sprayed on the wound surface. Chitosan was sprayed on the wound one minute after the solution did not flow out. After the wound was naturally formed, the wound in group B was only sprayed with EGF, and only in group C, only chitosan was sprayed on the wound. After its natural film formation, bandage and fixation. The wounds were treated again every 24 hours and the external dressing was changed. The gross morphology and healing of the wounds in the three groups were observed with naked eyes. Six rabbits were randomly selected for 3 days and 7 days and 10 days for 14 days. Six rabbits were killed by air embolization. The wound area was obtained by weighing paper and the wound healing rate was calculated. Under strict aseptic operation, the scab and tissue mass of scab and subeschar of scald wound were cut and cultured, and the total bacterial community per gram of wound tissue was calculated. The 100mg tissue of rabbit back wound was taken and the content of hydroxyproline was determined after treatment. The pathological indexes were observed after HE staining in some sections of the remaining specimens, and immunohistochemical indexes were observed after ki67 specific antibody staining in some specimens. Results: the wound in group A was less inflammatory than that in group B and C, the scab shell was soft and the wound shrank quickly. The rate of wound healing was significantly higher in group A than that in group B and C, but there was no significant difference between group A and group C. The histopathological specimens of the wound showed that the necrotic tissue attached loosely to the wound surface, the inflammatory cell chemotactic aggregation rate was faster, the endothelial cell proliferation was obvious, the collagen fiber proliferation was active, and the neonate epithelium was arranged neatly in group A than in group B and C. The positive rate of CFU / g in group A was lower than that in group B and group B was higher than that in group C and there was no significant difference between the two groups. The content of Hyp in group A was faster than that in group B (P < 0.05). Ki67 immunohistochemical analysis showed that the positive cells in group A were more compact, abundant and stained than those in group B. Ki67 immunohistochemical analysis showed that the positive cells in group A were more than those in group B and C, and the positive cells in group A were significantly higher than those in group B. There was no significant difference between the two groups (P < 0.05). The difference between group A and group C was significant. Conclusion: the combined use of the two drugs can enhance the healing ability of deep second degree scald in rabbits and make up for the defects of single drug. It can inhibit the growth of wound bacteria, accelerate the proliferation and growth of epithelial cells, and promote the synthesis of collagen.
【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R644

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