本文選題：液氮淺低溫冷凍 + 黑色素細胞��； 參考：《濱州醫(yī)學院》2010年碩士論文

【摘要】：目的: 觀察液氮淺低溫冷凍對家兔皮膚黑色素細胞及黑色素的組織學和超微結構的影響,以期為治療白癜風等色素脫失性疾病提供基礎理論依據。 方法: 1、實驗組采用棉簽蘸液氮法在家兔背部及臀部剃毛標記區(qū)反復涂擦三次(約1s),使局部結一層薄霜,待其自然融化后,再凍融2次為一周期,間隔一周再冷凍一周期,共3個周期。對照組和自身對照區(qū)同樣剪毛標記,不予冷凍。 2、冷凍后每天肉眼觀察皮膚組織的變化情況,包括色素的變化及毛發(fā)的生長情況等。 3、第3周和第5周取冷凍組和對照組標記區(qū)的皮膚組織,行HE染色和Masson-Fontana銀染色。利用Mias病理工作站4.0拍片觀察分析表皮和毛囊黑色素細胞數目及形態(tài)學變化、計數毛囊及真皮毛細血管數目;同時制備電鏡切片:觀察黑色素細胞及黑色素的超微結構變化。 4、統(tǒng)計學分析:應用SPSS13.0統(tǒng)計軟件,分別對第3周和第5周的數據進行t檢驗。 結果: 1、肉眼觀察:實驗區(qū)自第2周起部分區(qū)域出現片狀黑褐色色素沉著,并有灰黑色兔毛長出,較正常兔毛粗而黑,到第5周時色素沉著漸遍布整個冷凍標記區(qū),同時新生毛發(fā)長至正常長度。對照組:除2處標記區(qū)出現小片狀色素沉著斑及少許細軟、色淡的毳毛外,余標記區(qū)皮膚均未出現色素及新生兔毛。 2、HE和Masson-Fontana銀染色:實驗組表皮基底層和毛囊外根鞘處黑色素細胞數目和黑色素顆粒均明顯增加(P0.01),棘細胞層至顆粒層亦有黑色素顆粒沉積,真皮毛細血管數目增多(P0.01),管徑擴張顯著;第3周和第5周實驗組各相應項目指標行成組t檢驗,兩組四項相應指標的比較均無明顯統(tǒng)計學差異(P0.01)。 3、電鏡切片:實驗組:黑色素細胞胞質內線粒體較密集、粗面內質網和游離核糖體明顯增多,核切跡較少,核周有較多處于不同成熟階段的黑素小體,以Ⅱ和Ⅲ期為主,見少量Ⅳ期的黑素體,表皮細胞內含有大量的黑色素顆粒,以基底細胞及棘細胞為顯著。對照組:黑色素細胞胞核切跡較多,胞質內散在分布的內質網、核糖體、高爾基體及線粒體,胞核周圍有稀疏的處于不同成熟階段的黑色素體,以Ⅱ期為主;表皮各層中的黑色素顆粒較少,部分靠近棘層黑色素細胞的胞質內有成熟的黑色素顆粒。 結論: 1、液氮淺低溫冷凍能刺激表皮和毛囊的黑色素細胞使之數目增多、功能增強及促進多巴陰性的黑色素細胞合成黑色素,因而合成大量的黑色素;同時冷凍后真皮毛細血管數目增多、管徑擴張,促進了局部的血液循環(huán)和物質代謝,為新生毛發(fā)提供了營養(yǎng)基礎,因而新生毛發(fā)黑而粗,生長旺盛。 2、本實驗為治療白癜風等色素脫失性疾病提供了理論依據,而且液氮淺低溫冷凍療法有效率高,副作用少,值得臨床上廣泛應用。
[Abstract]:Objective: to observe the effect of liquid nitrogen cryopreservation on the histology and ultrastructure of melanocyte and melanin in rabbit skin in order to provide basic theoretical basis for the treatment of pigment loss diseases such as vitiligo. Methods: 1. In the experimental group, three times (about 1 s) were repeatedly applied to the rabbit's back and buttocks shaving marked area by using the cotton swabs dipping liquid nitrogen method, so that a thin layer of frost was formed locally, and then frozen and thawed twice for a period after the natural thawing. Once a week and then one cycle, a total of 3 cycles. The control group and the self-control area were also marked with haircut, without freezing. 2. After freezing, the changes of skin tissue were observed with the naked eye every day after freezing. The skin tissues of the frozen group and the control group were collected at the 3rd and 5th week and stained with HE and Masson-Fontana silver staining. The number and morphology of melanocytes in epidermis and hair follicle were observed and analyzed by Mias pathological workstation 4.0. The number of hair follicles and dermal capillaries were counted. The ultrastructural changes of melanocyte and melanin were observed. 4. Statistical analysis: using SPSS 13.0 statistical software, the data of the 3rd week and the 5th week were tested by t test. Results: 1. Naked eye observation: from the second week, some areas of the experimental area appeared patch dark brown pigmentation, and the gray and black hair grew out, which was thicker and darker than the normal rabbit hair. At the 5th week, the pigmentation gradually spread over the whole freezing labeling area. At the same time, the new hair grows to normal length. In the control group, except for 2 marked areas with small pigmented spots and a few soft, pale vellus hair, The number of melanocytes and melanin granules in the basal layer of epidermis and the outer root sheath of hair follicles in the experimental group were significantly increased (P0.01), and the layer of spinous cells to granules. The layer is also deposited with melanin particles, The number of dermal capillaries increased (P0.01). There was no significant difference in the four corresponding indexes between the two groups (P0.01). 3. Electron microscope section: in experimental group: mitochondria in melanocyte cytoplasm were denser, rough endoplasmic reticulum and free ribosome were increased obviously, and nuclear notch was less. There were many melanosomes in different maturation stages around the nucleus, mainly in stage 鈪，

本文編號：2083705