本文選題：殼聚糖 + 表皮生長(zhǎng)因子��； 參考：《瀘州醫(yī)學(xué)院》2013年碩士論文

【摘要】：目的：本實(shí)驗(yàn)通過離體細(xì)胞水平和建立兔深Ⅱ度燙傷自身對(duì)照模型，研究殼聚糖成膜噴劑聯(lián)合表皮細(xì)胞生長(zhǎng)因子噴霧劑（EGF）對(duì)兔深Ⅱ度燙傷創(chuàng)面的早中期的治療效果及其創(chuàng)面愈合的相關(guān)影響機(jī)制。通過觀察創(chuàng)面愈合的大體肉眼觀、創(chuàng)面愈合率的計(jì)算、痂下細(xì)菌計(jì)數(shù)、創(chuàng)面病理學(xué)觀察、創(chuàng)面組織羥脯氨酸(Hyp）含量變化、ki67在創(chuàng)面的分布和表達(dá)水平，探討聯(lián)合用藥在燙傷早中期治療的協(xié)同優(yōu)勢(shì)及其應(yīng)用前景。 方法：選取健康清潔級(jí)新西蘭大白兔24只，以兔脊柱為中間線在背部?jī)蓚?cè)用水燙法建立6個(gè)對(duì)稱的兔深Ⅱ度燙傷創(chuàng)面，上端兩個(gè)創(chuàng)面為實(shí)驗(yàn)組（A組：殼聚糖成膜劑聯(lián)合EGF組），中間兩個(gè)創(chuàng)面為對(duì)照組1（B組：EGF組），下端兩個(gè)創(chuàng)面為對(duì)照組2（C組：殼聚糖組），A組致傷后，先將EGF噴涂于創(chuàng)面，至藥液不外流為止，1分鐘后再將殼聚糖噴涂于創(chuàng)面，待其自然成膜后，，包扎固定。B組致傷后,僅在創(chuàng)面噴涂EGF至藥液不外流為止，包扎固定。C組致傷后，僅在創(chuàng)面噴涂殼聚糖，待其自然成膜后，包扎固定。每24h創(chuàng)面再次給藥，更換外敷料，肉眼觀察三組創(chuàng)面大體形態(tài)變化及愈合情況。選取3d、7d、10d、14d四個(gè)觀察時(shí)相點(diǎn)，每個(gè)時(shí)相點(diǎn)隨機(jī)抽取6只兔子，將抽取的6只兔子空氣栓塞處死，用稱量紙描繪計(jì)重得出創(chuàng)面面積并計(jì)算創(chuàng)面愈合率。在嚴(yán)格無菌操作下，切取燙傷創(chuàng)面焦痂及痂下組織塊1.0g，處理后行培養(yǎng)，計(jì)算每克創(chuàng)面組織的細(xì)菌群落總數(shù)。再取兔子背部創(chuàng)面100mg組織，處理后行組織羥脯氨酸（Hyp）含量測(cè)定。剩余的標(biāo)本組織一部分切片HE染色后觀測(cè)病理學(xué)指標(biāo)，一部分標(biāo)本行ki67特異性抗體染色后觀察免疫組化指標(biāo)。結(jié)果：創(chuàng)面大體觀察，A組創(chuàng)面較B、C兩組創(chuàng)面，創(chuàng)周炎性反應(yīng)輕，痂殼松軟，創(chuàng)面縮小速度快。創(chuàng)面愈合率的比較，A組明顯高于B、C兩組，B、C兩組之間的差異則不明顯。創(chuàng)面組織病理學(xué)標(biāo)本鏡下觀察見，A組創(chuàng)面較B、C兩組創(chuàng)面，壞死組織與創(chuàng)面附著松散，炎性細(xì)胞趨化聚集速度快，內(nèi)皮細(xì)胞增生明顯，膠原纖維增生活躍，新生上皮排列整齊。痂下細(xì)菌菌落數(shù)（CFU/g）陽性檢出率A組陽性檢出率低于B組，B組高于C組，A、B兩組差異不明顯。組織羥脯氨酸（Hyp）含量檢測(cè)，A組Hyp含量增長(zhǎng)速度快于B組，C組優(yōu)于B組，A、C兩組之間差異不大。ki67免疫組化分析，A組創(chuàng)面較B、C兩組創(chuàng)面，陽性細(xì)胞聚集緊密，量多，著色深，排列整齊。ki67指數(shù)陽性率比較，A、B兩組之間差異不明顯，A、B兩組與C組的差異均較明顯。結(jié)論：兩種藥物聯(lián)合使用能夠加強(qiáng)兔深Ⅱ度燙傷創(chuàng)面的愈合能力，并能彌補(bǔ)單一用藥的缺陷。能全面發(fā)揮其抑制創(chuàng)面細(xì)菌生長(zhǎng)，加快上皮細(xì)胞增殖生長(zhǎng)，促進(jìn)膠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.
【學(xué)位授予單位】：瀘州醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R644

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