羧甲基壳聚糖银对牙周炎的治疗作用
|
摘要
目的主要是:1、研究羧甲基壳聚糖银(CMCT-Ag+)对牙龈卟啉单胞菌(Pg)的抑制作用,为研制高效无毒的抗牙周病药物,提供实验依据。2、研究羧甲基壳聚糖银(Carboxymethyl Chitosan-Ag+, CMCT-Ag+)对实验性大鼠牙周炎的治疗作用。
方法:1、通过碱化、氯乙酸修饰和离子交换制备CMCT-Ag+。2、采用琼脂扩散和固体平板二倍稀释法测定CMCT-Ag+对Pg的抑制作用。3、以钢丝结扎大鼠右侧上颌第一磨牙、涂牙龈卟啉菌和饲喂高糖软饲料的方法构建实验性大鼠牙周炎模型。将12.5 mg,50 mg和200 mg/kg CMCT-Ag+涂于牙周病变部位;通过测量牙菌斑指数、牙龈出血指数和牙槽骨吸收值,研究CMCT-Ag+对实验性大鼠牙周炎的治疗作用。
结果:1、获得了粉红色的CMCT-Ag+粉末,其中银离子含量为10.21%。CMCT-Ag+的抑菌范围为0.3125-40 mg/ml,最低抑菌浓度为1.25 mg/ml。2、牙周炎组的出血指数(BI)、菌斑指数(PLI)及牙槽骨吸收水平(ABL)的均值与健康组比较有显著性差异(P<0.05);病理检查发现结合上皮向根方迁移,牙周袋形成,牙槽骨吸收,炎症细胞浸润,证明了牙周动物模型是成功的;羧甲基壳聚糖银组的出血指数、菌斑指数及牙槽骨吸收水平的均值与牙周炎组比较有显著性差异(P< 0.05)说明CMCT-Ag+对大鼠牙周炎有明显治疗作用,其疗效与替硝唑相似,但对于牙槽骨破坏的修复无明显直接改善作用。
结论:1、CMCT-Ag+对Pg的生长有明显抑制作用,可用于进一步研制治疗牙周炎药物。2、CMCT-Ag+对实验性大鼠牙周炎有一定治疗作用,其作用机制主要是抑制牙周组织中细菌的发生。
Periodontal disease was one of prevalence and liability oral diseases. Gingivitis and periodontitis were chiefly two kinds of periodontal diseases. Periodontal disease was a group of ailments that occurred in periodontal tissue. Its results were junction epithelium migration to the dental root, periodontal pocket formation, tooth mobility or displace. Serious Periodontal diseases can cause loss of tooth. Now, in the word, there isn't better method on treating the diseases.
Bacteria infection, especially the anaerobic bacteria, was the initiating factor of periodontal diseases, so how to efficiently control the infection was the mainly objective of treatment and prevention periodontal diseases in clinic. At present, there are two methods to eliminate or decrease pathogenic bacteria. One is to remove all the dental plaque and calculus adhered to the teeth surface by using mechanical tools in order to decrease the amount of bacteria around teeth. However, the working rationale and character are different from all ultrasonic scaler and their effect and advantages or disadvantages are seldom compared and estimated in our country by special clinical research. These results will bring some difficulties on the selection of different scaler for clinical use. The other is to control the periodontal anaerobic bacteria by using drugs such as metronidazole and tinidazole, the most widely used drugs particularly anti-anaerobic bacteria drugs. Because oral application drugs must be used higher dosage, it may bring some side effects such as nausea, liver damage, anorectic reaction, etc. Local drugs can also overcome these shortcomings,so local sustained-delivery drugs will be widely applied. It have become great focus in the recent research.
Chitosan, the disacetylizad derivative of chitin, was the only alkaline polysaccharide in nature. Special chemical structure brought it interesting bioactivities, such as antimicrobial, anti-tumor, immune enhancing effect and tissue regenerative activities. It was widely used in medical field due to its additional biocompatibilily and biodegradability. Chitosan could be easily made various dosage form to be applied in various drug delivery pathways. In addition, its special bio-adhesive property help to prevent the rapid elimination caused by the flushing action of saliva. As an important derivative of Chitosan, Silver Carboxymethyl Chitosan (CMCT-Ag+) owned sterilizing ability besides of the characteristic bioactivities and biocompatibility of Chitosan. It was an ideal sustained-delivery drug. CMCT-Ag+ had great application potential in preventing and treating periodontal.
In this study, we synthesized CMCT-Ag+ by alkalization, modification of chloracetic acid and ion exchange. Then, we observed the effect of CMCT-Ag+ against Porphyromonas gingivalis(Pg) by the agar diffusion and plate serial dilution test. We set up periodontitis model in rats. Then, the CMCT-Ag+ therapeutic effects to periodontitis rats were detected by the PLI, BI and ABL. The results indicated that CMCT-Ag+ was pink powder, and the content of Ag+ was 10.21 % in the obtained CMCT-Ag+. The inhibitory concentration of CMCT-Ag+ ranged from 0.31 mg/ml to 40 mg/ml, and the minimal inhibitory concentration (MIC) was 1.25 mg/ml. In the CMCT-Ag+ group, PLI and BI were significantly improved compared with the Periodontitis group (P<0.05). But the repair of alveolar bone loss was not obviously. By pathological observation, we discovered that Periodontitis group had some changes of inflammatory cell infiltration, alveolar bone loss and periodontal pocket formation. In CMCT-Ag+ group, inflammatory cell infiltration was obviously release, periodontal pocket transformed superficial and dento-soft tissue was obviously repaired, but alveolar bone loss still exist. These results showed that CMCT-Ag+ could significantly treat rats periodontitis, but the destruction of alveolar bone was not recovered in short time.
Our results showed that the CMCT-Ag+ could inhibit the growth of Pg, treat rats periodontitis by inhibition periodontal bacteria. This study provided theoretical and experimental foundation for prepaing local drug of sustained-delivery systems and developing higher efficiency and reducing side-effect drug to treat periodontal disease.
方法:1、通过碱化、氯乙酸修饰和离子交换制备CMCT-Ag+。2、采用琼脂扩散和固体平板二倍稀释法测定CMCT-Ag+对Pg的抑制作用。3、以钢丝结扎大鼠右侧上颌第一磨牙、涂牙龈卟啉菌和饲喂高糖软饲料的方法构建实验性大鼠牙周炎模型。将12.5 mg,50 mg和200 mg/kg CMCT-Ag+涂于牙周病变部位;通过测量牙菌斑指数、牙龈出血指数和牙槽骨吸收值,研究CMCT-Ag+对实验性大鼠牙周炎的治疗作用。
结果:1、获得了粉红色的CMCT-Ag+粉末,其中银离子含量为10.21%。CMCT-Ag+的抑菌范围为0.3125-40 mg/ml,最低抑菌浓度为1.25 mg/ml。2、牙周炎组的出血指数(BI)、菌斑指数(PLI)及牙槽骨吸收水平(ABL)的均值与健康组比较有显著性差异(P<0.05);病理检查发现结合上皮向根方迁移,牙周袋形成,牙槽骨吸收,炎症细胞浸润,证明了牙周动物模型是成功的;羧甲基壳聚糖银组的出血指数、菌斑指数及牙槽骨吸收水平的均值与牙周炎组比较有显著性差异(P< 0.05)说明CMCT-Ag+对大鼠牙周炎有明显治疗作用,其疗效与替硝唑相似,但对于牙槽骨破坏的修复无明显直接改善作用。
结论:1、CMCT-Ag+对Pg的生长有明显抑制作用,可用于进一步研制治疗牙周炎药物。2、CMCT-Ag+对实验性大鼠牙周炎有一定治疗作用,其作用机制主要是抑制牙周组织中细菌的发生。
Periodontal disease was one of prevalence and liability oral diseases. Gingivitis and periodontitis were chiefly two kinds of periodontal diseases. Periodontal disease was a group of ailments that occurred in periodontal tissue. Its results were junction epithelium migration to the dental root, periodontal pocket formation, tooth mobility or displace. Serious Periodontal diseases can cause loss of tooth. Now, in the word, there isn't better method on treating the diseases.
Bacteria infection, especially the anaerobic bacteria, was the initiating factor of periodontal diseases, so how to efficiently control the infection was the mainly objective of treatment and prevention periodontal diseases in clinic. At present, there are two methods to eliminate or decrease pathogenic bacteria. One is to remove all the dental plaque and calculus adhered to the teeth surface by using mechanical tools in order to decrease the amount of bacteria around teeth. However, the working rationale and character are different from all ultrasonic scaler and their effect and advantages or disadvantages are seldom compared and estimated in our country by special clinical research. These results will bring some difficulties on the selection of different scaler for clinical use. The other is to control the periodontal anaerobic bacteria by using drugs such as metronidazole and tinidazole, the most widely used drugs particularly anti-anaerobic bacteria drugs. Because oral application drugs must be used higher dosage, it may bring some side effects such as nausea, liver damage, anorectic reaction, etc. Local drugs can also overcome these shortcomings,so local sustained-delivery drugs will be widely applied. It have become great focus in the recent research.
Chitosan, the disacetylizad derivative of chitin, was the only alkaline polysaccharide in nature. Special chemical structure brought it interesting bioactivities, such as antimicrobial, anti-tumor, immune enhancing effect and tissue regenerative activities. It was widely used in medical field due to its additional biocompatibilily and biodegradability. Chitosan could be easily made various dosage form to be applied in various drug delivery pathways. In addition, its special bio-adhesive property help to prevent the rapid elimination caused by the flushing action of saliva. As an important derivative of Chitosan, Silver Carboxymethyl Chitosan (CMCT-Ag+) owned sterilizing ability besides of the characteristic bioactivities and biocompatibility of Chitosan. It was an ideal sustained-delivery drug. CMCT-Ag+ had great application potential in preventing and treating periodontal.
In this study, we synthesized CMCT-Ag+ by alkalization, modification of chloracetic acid and ion exchange. Then, we observed the effect of CMCT-Ag+ against Porphyromonas gingivalis(Pg) by the agar diffusion and plate serial dilution test. We set up periodontitis model in rats. Then, the CMCT-Ag+ therapeutic effects to periodontitis rats were detected by the PLI, BI and ABL. The results indicated that CMCT-Ag+ was pink powder, and the content of Ag+ was 10.21 % in the obtained CMCT-Ag+. The inhibitory concentration of CMCT-Ag+ ranged from 0.31 mg/ml to 40 mg/ml, and the minimal inhibitory concentration (MIC) was 1.25 mg/ml. In the CMCT-Ag+ group, PLI and BI were significantly improved compared with the Periodontitis group (P<0.05). But the repair of alveolar bone loss was not obviously. By pathological observation, we discovered that Periodontitis group had some changes of inflammatory cell infiltration, alveolar bone loss and periodontal pocket formation. In CMCT-Ag+ group, inflammatory cell infiltration was obviously release, periodontal pocket transformed superficial and dento-soft tissue was obviously repaired, but alveolar bone loss still exist. These results showed that CMCT-Ag+ could significantly treat rats periodontitis, but the destruction of alveolar bone was not recovered in short time.
Our results showed that the CMCT-Ag+ could inhibit the growth of Pg, treat rats periodontitis by inhibition periodontal bacteria. This study provided theoretical and experimental foundation for prepaing local drug of sustained-delivery systems and developing higher efficiency and reducing side-effect drug to treat periodontal disease.
引文
[1] 张翼. 对牙周疾病认识的辩证思维[J]. 医学与社会, 2006,19(7): 27-29.
[2] 黎 炜 . 牙 周 炎 的 难 治 性 原 因 [J]. 临 床 和 实 验 医 学 杂 志 , 2006,5(10):1603.
[3] 郭永华,吴亚菲,刘天佳. 牙龈卟啉单胞菌侵入牙龈上皮细胞的分子机制[J]. 国外医学口腔医学分册,2005,32(2): 90-92.
[4] 葛少华. 牙周病的病因及发病机制[J]. 山东医药, 2001,41(9): 49-51.
[5] 韦名浪,何克新. 牙周病药物治疗现状[J]. 右江医学, 2004,32(5): 493-495.
[6] 顾晶晶,胡纯贞,王蓓. 替硝唑治疗牙周炎的临床疗效观察[J]. 口腔材料器械杂志, 1998,7(3)∶142-143.
[7] Kinane DF, Radvar MA. Six-month comparison of three periodontal local antimicrobial therapies in persistent periodontal pockets. J Periodontol, 1999,70(1):1-7.
[8] Park YJ, Lee YM, Park SN et al. Enhance guide bone regeneration by controlled tetracycline release from poly(L-lactied) barrier membrans. J Biomed Mater Res,2000,51(3): 391-397.
[9] Feres M, Haffajee AD, Goncalves C et al. Systemic doxycycline adminstration in the treatment of periodontal infections(Ⅱ) effect on antibiotic resistance of subgingival species. J Clin Periogontol ,1999,26(5): 784-792.
[10] Steenberghe D, Rosling B, Soder P et al. A 15-month evalution of the effects of repeated subgingival minocycline in chronic adult periodontitis. J Periodontol, 1999,70(6): 657-667.
[11] 邓嘉胤,彭诚. 洁治术后服替硝唑和乙酰螺旋霉素治疗成人牙周炎疗效观察[J]. 天津医科大学学报, 2000,6 (1)∶97-98.
[12] 吴亚菲,赵蕾. 牙周病药物治疗及进展[J]. 中国医药刊, 2006,8(4): 285-288.
[13] Buduneli N,Vardar S, Atilla G et al. Gingival crevicular fluid matrix metalloproteinase-8 levels following adjunctive use of meloxicam and initial phase of periodontal therapy. J Periodontol, 2002,73(1): 103-109.
[14] 徐叔云,熊祖应,丁长海. 抗炎免疫药理学研究进展[J]. 药理学新进展, 2000: 37-46.
[15] 徐叔云. 抗炎免疫药物学研究十年进展. 中国药理学通报, 1996,12(1): 1-6.
[16] Cavanaugh Jr PF, Meredith MP, Buchanan W et al. Coordinate production of PGE2 and IL-1β in the gingival crevicular fluid of adults with periodontitis : Its relationship to alveolar bone loss and disruption by twice daily treatment with ketorolac tromethamine oral rinse. J Periodont Res , 1998,33(1): 75-82.
[17] Lima V, Bezerra V, Alencar DM et al. Effects of chlorpromazine on alveolar bone loss in experimental periodontal disease in rats. Eur J Oral Sci, 2000,108 (2): 123-129.
[18] 张举之. 固齿丸治疗牙周病机理探讨[J]. 华西口腔医学杂志, 1990,8(3): 198-200.
[19] 宋红, 赵瑞芳, 周以钧. 固齿膏对牙周炎龈沟液中 IL-8 的影响[J]. 中华口腔医学杂志, 1997,32(6): 373- 374.
[20] 吴敏,陈少武. 三种龈下冲洗液对成人牙周炎辅助治疗的疗效比较[J]. 口腔医学研究, 2003,19(4): 316- 317.
[21] 葛久禹,李振汉,平蕾,等. 西吡氯铵漱口液对牙龈炎、牙周炎疗效的临床评价[J]. 牙体牙髓牙周病学杂志, 2002,12(7)∶ 373-377.
[22] 孙冀平. 壳聚糖及其应用[J]. 中国食品添加剂, 2005, (5): 83-86.
[23] 刘琨,侯本祥,杨圣辉,等. 壳聚糖体外抑菌实验研究[J] . 现代口腔医学杂志, 2007,21(3): 281-283.
[24] 蒋玉燕, 毕忆群. 275 株病原菌对壳聚糖的敏感性试验[J]. 浙江省医学科学院学报, 1995,16(2): 7-9.
[25] 吕键,刘立春,池善女,袁勇,崔胜云. 甲壳素、壳聚糖及其衍生物在医药领域的应用[J]. 延边大学学报(自然科学版), 2003,29(4): 261-267.
[26] 郑连英, 朱江峰, 孙昆山.壳聚糖的抗菌性能研究[J]. 材料科学与工程, 2000,18(2):22-24.
[27] Muzzarelli R, Jeuaiaux C, Gooday GW. Chitin in nature and Technology[M]. New York: Plenum Press, 1985,210.
[28] Sano H, Shibasaki K, Matsukubo T, et al. Effect of chitosan rinsing on reduction of dental p laque formation[J]. Bull Tokyo Dent Coll, 2003,44(1): 9-16.
[29] 陈薇,杨圣辉,果梅英. 甲壳胺复合物影响变形链球菌生长、代谢的实验室研究[J]. 现代口腔医学杂志, 2004, 18(5): 292-295.
[30] Ikinci G, Senel S , Akincibay H et al. Effect of chitosan on aperiodontal pathogen Porphyromonas gingivalis. Int J. Pharm. 2002,235(122): 121.
[31] Tarsi R, Corbin B, Pruzzo C, et a1. Effect of low molecular weight chitosans on the adhesive p roperties of oral strep tococci[J]. Oral Microbial Immunol, l998,13(4): 217-224.
[32] Tarsi R, Muzzarelli RA, Guzman CA, et a1. Inhibition of Strep tococcus mutans adsorp tion to hydroxyapatite by low molecular weight chitosans[J]. J Den Res, 1997,76(2): 665–672.
[33] 付田霞,刘晓宇,刘志恒. 壳聚糖的抑菌性能应用研究进展[J]. 辽宁化工, 2005,34(12): 541-543.
[34] 刘瑾,唐梓进. 羧甲基壳聚糖对口腔变形链球菌和口腔乳酸杆菌的抑制作用[J]. 南京师范大学学报(自然科学版), 1999,22(4): 731.
[35] 唐涛,薛毅,信玉华,等. 羧甲基壳聚糖对口腔重要厌氧菌的抑菌性能评价[J]. 中国微生态学杂志, 2003,15(6): 337-339.
[36] 赵丽瑞,孙多先,刘满英. 羧甲基壳聚糖的性能及其在 生物医学领域的应用[J]. 高分子通报, 2007,8: 43-47.
[37] 刘瑾,唐梓进,杨佐宪. 唾液腺素、羧甲基壳聚糖、低分子壳聚糖对豚鼠齿龈炎的疗效[J]. 南京师大学报(自然科学版),2000,23(2):103-104.
[38] 夏金兰,王春,刘新星. 抗菌剂及其抗菌机理[J]. 中南大学学报(自然科学版), 2004,35(1): 31-38.
[39] 湛学军,熊远珍,谢大泽. 羧甲基壳聚糖银的合成及抑菌实验的研究[J]. 中国生化药物杂志, 2001,22(3): 142-144.
[40] 李霞,程珏,孙克勤,等. 结扎丝法构建大鼠实验性牙周炎模型的研究[J]. 首都医药,2007,7:34-35.
[41] 党诚学,陈松旺,郭先予,等. 凝集素受体、ras P21的表达,AgNOR和性激素受体在乳腺肿瘤诊断和预后中的意义[J]. 现代肿瘤临床杂志,1993,1(1):16-17
[42] 李克安,曾伟,童沈阳. 络合物溶液中各物种平衡浓度的计算[J]. 计算机与应用化学,1989,6(1):35-40.
[43] 张文钲. 银的杀菌功能[J]. 金属世界,2002,3:20-21.
[44] 汤绚丽,姚根有,陈丽荣,等. 乳腺癌及癌前病变3号染色体短臂杂合性缺失的研究[J]. 中华外科杂志,2006,44(19):1314-1317.
[45] 湛学军,熊远珍,柳哲,等. 羧甲基壳聚糖银的合成及抑菌实验的研究[J], 中国生化药物杂志,2001,22(3):142-144.
[46] 赵玉清,刘宝全,李慧, 等. 壳聚糖-Ag(I)的配位与抑菌性研究[J]. 北华大学学报(自然科学版),2000,1(5): 384-386.
[47] 曹采方主编. 牙周病学(第4版)[M]. 人民卫生出版社,2000,38-47.
[48] Clickman. Clinical periodontology,fourth edition. Saudders WB. Company. USA.1999, 291-415 .
[49] Achong R, Nishimura I, Ramachandran H, et al. Membrane type(MT) 1 - matrix metallop roteinase (MMP) and MMP - 2 expression in ligature - induced periodontitis in the rat[J]. J Periodontol, 2003, 74 (4) : 494 – 500.
[50] Schreiner HC, Sinatra K, Kap lan JB, et al. Tight– adherence genes of Actinobacillus actinomycetem comitans are required for virulence in a rat model[J]. Proc N atl Acad Sci USA, 2003, 100(12) : 7295 - 7300
[51] Fischer RG, King B. Clinical and histological evalluation of ligture-induced periodontitis in the donestic ferret. J Clin Perionont, 1994,21: 230 .
[52] 陈铁楼,等. 高压氧对豚鼠实验性牙周炎作用的组织病理学研究 [J]. 口腔医学杂志,2002,18(5): 418-420.
[53] 施化莲,等. 可降解缓释药膜对大鼠牙周炎的治疗作用[J]. 生物医学工程与临床, 1998,2(2): 88-89 .
[54] 王琳辉,季晖,任萍,等. 替硝唑口腔贴膜对实验性大鼠牙周炎的治疗作用及其对口腔粘膜刺激性研究[J]. 中国临床药理学与治疗学, 2004,9(1): 39-43.
[2] 黎 炜 . 牙 周 炎 的 难 治 性 原 因 [J]. 临 床 和 实 验 医 学 杂 志 , 2006,5(10):1603.
[3] 郭永华,吴亚菲,刘天佳. 牙龈卟啉单胞菌侵入牙龈上皮细胞的分子机制[J]. 国外医学口腔医学分册,2005,32(2): 90-92.
[4] 葛少华. 牙周病的病因及发病机制[J]. 山东医药, 2001,41(9): 49-51.
[5] 韦名浪,何克新. 牙周病药物治疗现状[J]. 右江医学, 2004,32(5): 493-495.
[6] 顾晶晶,胡纯贞,王蓓. 替硝唑治疗牙周炎的临床疗效观察[J]. 口腔材料器械杂志, 1998,7(3)∶142-143.
[7] Kinane DF, Radvar MA. Six-month comparison of three periodontal local antimicrobial therapies in persistent periodontal pockets. J Periodontol, 1999,70(1):1-7.
[8] Park YJ, Lee YM, Park SN et al. Enhance guide bone regeneration by controlled tetracycline release from poly(L-lactied) barrier membrans. J Biomed Mater Res,2000,51(3): 391-397.
[9] Feres M, Haffajee AD, Goncalves C et al. Systemic doxycycline adminstration in the treatment of periodontal infections(Ⅱ) effect on antibiotic resistance of subgingival species. J Clin Periogontol ,1999,26(5): 784-792.
[10] Steenberghe D, Rosling B, Soder P et al. A 15-month evalution of the effects of repeated subgingival minocycline in chronic adult periodontitis. J Periodontol, 1999,70(6): 657-667.
[11] 邓嘉胤,彭诚. 洁治术后服替硝唑和乙酰螺旋霉素治疗成人牙周炎疗效观察[J]. 天津医科大学学报, 2000,6 (1)∶97-98.
[12] 吴亚菲,赵蕾. 牙周病药物治疗及进展[J]. 中国医药刊, 2006,8(4): 285-288.
[13] Buduneli N,Vardar S, Atilla G et al. Gingival crevicular fluid matrix metalloproteinase-8 levels following adjunctive use of meloxicam and initial phase of periodontal therapy. J Periodontol, 2002,73(1): 103-109.
[14] 徐叔云,熊祖应,丁长海. 抗炎免疫药理学研究进展[J]. 药理学新进展, 2000: 37-46.
[15] 徐叔云. 抗炎免疫药物学研究十年进展. 中国药理学通报, 1996,12(1): 1-6.
[16] Cavanaugh Jr PF, Meredith MP, Buchanan W et al. Coordinate production of PGE2 and IL-1β in the gingival crevicular fluid of adults with periodontitis : Its relationship to alveolar bone loss and disruption by twice daily treatment with ketorolac tromethamine oral rinse. J Periodont Res , 1998,33(1): 75-82.
[17] Lima V, Bezerra V, Alencar DM et al. Effects of chlorpromazine on alveolar bone loss in experimental periodontal disease in rats. Eur J Oral Sci, 2000,108 (2): 123-129.
[18] 张举之. 固齿丸治疗牙周病机理探讨[J]. 华西口腔医学杂志, 1990,8(3): 198-200.
[19] 宋红, 赵瑞芳, 周以钧. 固齿膏对牙周炎龈沟液中 IL-8 的影响[J]. 中华口腔医学杂志, 1997,32(6): 373- 374.
[20] 吴敏,陈少武. 三种龈下冲洗液对成人牙周炎辅助治疗的疗效比较[J]. 口腔医学研究, 2003,19(4): 316- 317.
[21] 葛久禹,李振汉,平蕾,等. 西吡氯铵漱口液对牙龈炎、牙周炎疗效的临床评价[J]. 牙体牙髓牙周病学杂志, 2002,12(7)∶ 373-377.
[22] 孙冀平. 壳聚糖及其应用[J]. 中国食品添加剂, 2005, (5): 83-86.
[23] 刘琨,侯本祥,杨圣辉,等. 壳聚糖体外抑菌实验研究[J] . 现代口腔医学杂志, 2007,21(3): 281-283.
[24] 蒋玉燕, 毕忆群. 275 株病原菌对壳聚糖的敏感性试验[J]. 浙江省医学科学院学报, 1995,16(2): 7-9.
[25] 吕键,刘立春,池善女,袁勇,崔胜云. 甲壳素、壳聚糖及其衍生物在医药领域的应用[J]. 延边大学学报(自然科学版), 2003,29(4): 261-267.
[26] 郑连英, 朱江峰, 孙昆山.壳聚糖的抗菌性能研究[J]. 材料科学与工程, 2000,18(2):22-24.
[27] Muzzarelli R, Jeuaiaux C, Gooday GW. Chitin in nature and Technology[M]. New York: Plenum Press, 1985,210.
[28] Sano H, Shibasaki K, Matsukubo T, et al. Effect of chitosan rinsing on reduction of dental p laque formation[J]. Bull Tokyo Dent Coll, 2003,44(1): 9-16.
[29] 陈薇,杨圣辉,果梅英. 甲壳胺复合物影响变形链球菌生长、代谢的实验室研究[J]. 现代口腔医学杂志, 2004, 18(5): 292-295.
[30] Ikinci G, Senel S , Akincibay H et al. Effect of chitosan on aperiodontal pathogen Porphyromonas gingivalis. Int J. Pharm. 2002,235(122): 121.
[31] Tarsi R, Corbin B, Pruzzo C, et a1. Effect of low molecular weight chitosans on the adhesive p roperties of oral strep tococci[J]. Oral Microbial Immunol, l998,13(4): 217-224.
[32] Tarsi R, Muzzarelli RA, Guzman CA, et a1. Inhibition of Strep tococcus mutans adsorp tion to hydroxyapatite by low molecular weight chitosans[J]. J Den Res, 1997,76(2): 665–672.
[33] 付田霞,刘晓宇,刘志恒. 壳聚糖的抑菌性能应用研究进展[J]. 辽宁化工, 2005,34(12): 541-543.
[34] 刘瑾,唐梓进. 羧甲基壳聚糖对口腔变形链球菌和口腔乳酸杆菌的抑制作用[J]. 南京师范大学学报(自然科学版), 1999,22(4): 731.
[35] 唐涛,薛毅,信玉华,等. 羧甲基壳聚糖对口腔重要厌氧菌的抑菌性能评价[J]. 中国微生态学杂志, 2003,15(6): 337-339.
[36] 赵丽瑞,孙多先,刘满英. 羧甲基壳聚糖的性能及其在 生物医学领域的应用[J]. 高分子通报, 2007,8: 43-47.
[37] 刘瑾,唐梓进,杨佐宪. 唾液腺素、羧甲基壳聚糖、低分子壳聚糖对豚鼠齿龈炎的疗效[J]. 南京师大学报(自然科学版),2000,23(2):103-104.
[38] 夏金兰,王春,刘新星. 抗菌剂及其抗菌机理[J]. 中南大学学报(自然科学版), 2004,35(1): 31-38.
[39] 湛学军,熊远珍,谢大泽. 羧甲基壳聚糖银的合成及抑菌实验的研究[J]. 中国生化药物杂志, 2001,22(3): 142-144.
[40] 李霞,程珏,孙克勤,等. 结扎丝法构建大鼠实验性牙周炎模型的研究[J]. 首都医药,2007,7:34-35.
[41] 党诚学,陈松旺,郭先予,等. 凝集素受体、ras P21的表达,AgNOR和性激素受体在乳腺肿瘤诊断和预后中的意义[J]. 现代肿瘤临床杂志,1993,1(1):16-17
[42] 李克安,曾伟,童沈阳. 络合物溶液中各物种平衡浓度的计算[J]. 计算机与应用化学,1989,6(1):35-40.
[43] 张文钲. 银的杀菌功能[J]. 金属世界,2002,3:20-21.
[44] 汤绚丽,姚根有,陈丽荣,等. 乳腺癌及癌前病变3号染色体短臂杂合性缺失的研究[J]. 中华外科杂志,2006,44(19):1314-1317.
[45] 湛学军,熊远珍,柳哲,等. 羧甲基壳聚糖银的合成及抑菌实验的研究[J], 中国生化药物杂志,2001,22(3):142-144.
[46] 赵玉清,刘宝全,李慧, 等. 壳聚糖-Ag(I)的配位与抑菌性研究[J]. 北华大学学报(自然科学版),2000,1(5): 384-386.
[47] 曹采方主编. 牙周病学(第4版)[M]. 人民卫生出版社,2000,38-47.
[48] Clickman. Clinical periodontology,fourth edition. Saudders WB. Company. USA.1999, 291-415 .
[49] Achong R, Nishimura I, Ramachandran H, et al. Membrane type(MT) 1 - matrix metallop roteinase (MMP) and MMP - 2 expression in ligature - induced periodontitis in the rat[J]. J Periodontol, 2003, 74 (4) : 494 – 500.
[50] Schreiner HC, Sinatra K, Kap lan JB, et al. Tight– adherence genes of Actinobacillus actinomycetem comitans are required for virulence in a rat model[J]. Proc N atl Acad Sci USA, 2003, 100(12) : 7295 - 7300
[51] Fischer RG, King B. Clinical and histological evalluation of ligture-induced periodontitis in the donestic ferret. J Clin Perionont, 1994,21: 230 .
[52] 陈铁楼,等. 高压氧对豚鼠实验性牙周炎作用的组织病理学研究 [J]. 口腔医学杂志,2002,18(5): 418-420.
[53] 施化莲,等. 可降解缓释药膜对大鼠牙周炎的治疗作用[J]. 生物医学工程与临床, 1998,2(2): 88-89 .
[54] 王琳辉,季晖,任萍,等. 替硝唑口腔贴膜对实验性大鼠牙周炎的治疗作用及其对口腔粘膜刺激性研究[J]. 中国临床药理学与治疗学, 2004,9(1): 39-43.