影響義齒樹脂表面對白色念珠菌黏附的因子

Factors Influenced Candida Albicans Adhesion on Denture Surface

洪嘉伶(Chia-Ling Hung)

指導教授 ： 王若松

共同指導教授 ： 林立德

國立臺灣大學/牙醫專業學院/臨床牙醫學研究所/碩士(2009年)

https://doi.org/10.6342/NTU.2009.02778

摘要

口腔中義齒的存在是造成義齒性口炎的最主要因素，而白色念珠菌最常在義齒殘留的牙菌斑中分離出來。義齒的表面粗度在顯微鏡下常可見到細微的裂隙或不均勻孔洞，一般認為樹脂的粗糙度是黏附的重要因子。另外也有學者模擬白色念珠菌在口中的黏附黏膜的機制做探討，但至今結果仍未有定論。在以往的研究上都偏向於物理性質的探討，而關於一些後續的因子並未有相互影響的比較。
本實驗的目的要於評估三種可能會影響白色念珠菌對義齒黏附的因子，分別是製作過程中所塗布的分離劑、拋光的處理以及唾液的影響。
實驗材料與方法: 用臨床上製作標準活動義齒的程序製備樹脂試片148片(一半是經過分離劑處理，一半是沒有經過分離劑處理)，大小為20x50x2mm。這兩大組再以拋光有無以及浸泡唾液有無隨機分為四組(總共八組)。每組皆利用表面粗糙度測定器Surftest4測量表面粗糙度參數(Ra)後，再開始進行黏附實驗。所有試片皆以超音波震盪器洗淨消毒後，在無菌的操作技術下將其浸泡在含有濃度約105 cell/ml 的白色念珠菌(SC5314)的菌液或是唾液加菌液中。一小時後，將所有試片取出在室溫下陰乾，並以固定力量印壓在培養基上(Sabourand dextrose agar)半小時後拿掉。紀錄其培養基培養18小時後所形成的菌落數。所有的資料結果以ANOVA及多變數回歸分析了解各因子的影響力。
結果: 在白色念珠菌黏附與三個影響因子的相關度: 可以發現拋光與塗布分離劑對白色念珠菌附著的影響為正相關，而對表面粗糙度參數Ra為負相關。口水則對白色念珠球菌黏附影響呈現負相關。此外，在白色念珠菌黏附度，分離劑的影響度比對拋光來的大。回歸分析的結果也顯示，塗布分離劑對於白色念珠菌落形成(Colony forming unit, CFU)有達到顯著的增加(P=0.0008)，但拋光動作則沒有達到統計上的顯著。唾液的影響則是會減少白色念珠菌的黏附(P=0.001)。將所有組的Ra與CFU做一個圖表分析，可以發現所有的數值幾乎集中在Ra為1~3μm.之間。而在光顯與電顯的觀察下，似乎可發現在有塗布分離劑組的樹脂表面，有較均勻且成群的白色念珠菌，且常可以發現菌絲的生長，而不管在哪一組，白色念珠球菌皆較會黏附在表面凹凸不平的平面或孔洞內。
結論:我們的實驗結果顯示樹脂片製作過程中，塗布分離劑似乎對於白色念珠球菌黏附的影響力比拋光動作來的大(正向影響)，而唾液似乎會減少白色念珠菌的黏附。

關鍵字

白色念珠菌黏附 ； 口水 ； 義齒表面粗糙度 ； 分離劑

並列摘要

Candida albicans was the most common yeast species found in denture plaque of patients with denture stomatitis. Regarding to Candida albicans adhesion on denture surface, many studies had focused on the influences of physical characteristics of acrylic resin, such as surface roughness, surface free energy or resin materials, etc… The denture fabrication process is complicated which may also play an important role in Candida adhesion. Moreover, denture is soaked in saliva all the time during wearing. Saliva is viscous and contains enzymes which may complicate the adhesion of Candida on denture surface. Seldom studies focused on factors of denture fabrication procedures and saliva together to evaluate the adhesion of Candida albicans on denture surface.
The aim of this in vitro study was to evaluate the relationship between factors that may influence adhesion of Candida albicans on denture surface, such as denture surface roughness, separating fluid used in denture processing and human saliva.
Materials and methods:
148 pieces of heat-cured acrylic resin (Luciton 199), 20x50x2mm in size, were fabricated by standard procedure. Saliva was obtained from 5 different health people. The specimens were randomly separated into 8 groups which were treated with three different factors during resin plate fabrication procedure: with- or without separating fluid, with or without polishment procedure, immerse in saliva or not. Each group has its negative control. The surface roughness parameters (Ra) of all resin plates were measured by Surftest4 before the study. After cleaning and disinfection, all samples were placed into Candida albicans SC5314 broth with concentration of 105 cell/ml. The test tube was incubated for 1 hour at 37oC in an orbital shaker to promote yeast adherence. On hour later, all samples were air dried and imprinted on SDA agar for half hour. Then the resin plates were removed and agar was incubated at 37oC for 18 hours. Colony forming unit (CFU) of Candida Albicans on the agar surface were counted and the results were analyzed by correlation analysis, multiple regression analysis & ANOVA.
Result:
The results showed that surface roughness in this study was not a significant factor influenced the Candida albicans adhesion on denture surface. (P=0.562). Using separating fluid during acrylic resin processing was significant which increased Candida adhesion (P=0.008) Human saliva was also showed to be a significant factor but would decrease the Candida adhesion (P=0.001), There was no interaction among these experiment factors.
Conclusion:
It can be concluded that denture separating fluid and human saliva showed influences on Candida adhesion, the former is positive, and the later is negative.

並列關鍵字

Candida adhesion ； Surface roughness ； saliva ； denture separating fluid

參考文獻

43. Lin CH, Chen TC, Lin LD, Wang TW, Wang JS, The influence of denture surface roughness on the adhesion of Candida albicans. Formosan Journal of Medicine, 2005. 9: p. 332-8.

2. Crumming CG, Weight C, Blackwell CL, Denture stomatitis in the eldly. Oral Microbiology and Immunology, 1990. 5: p. 82-85.

4. Budtz-Jorgensen E, Bertram U, Denture stomatitis vs. candida agglutinins in human serum. Acta Odontologica Scandinavica, 1972. 30: p. 313.

5. Grabowaki, IM , Bertram U, Oral Health status and need of dental treatment in the elderly Spanish population. Community Dentistry and Oral Epidemiology, 1975. 3: p. 108.

6. Adrendorf TM, Walker DM, Denture stomatitis: a review. Journal of Oral Rehabilitation, 1987. 14: p. 217-227.