牙周可疑致病微生物与种植体周病变的相关性研究
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摘要
目的：
1．检测健康及炎症种植体龈下菌斑中7种牙周可疑致病微生物的定殖情况；
2．探讨牙周可疑致病微生物与种植体周围炎症及骨吸收的关系；
3．探讨种植体周病变的微生物学病因，寻找能早期反映种植体周病变的主要致病微生物；
4．探索早期诊断及早期防治种植体周围炎的方法。
方法：
1．病例收集分组：选择种植修复后12个月以上的88位患者的89颗种植体，分三组①种植体周围炎组29例29颗种植体周围炎患者、②种植体周粘膜炎组29例30颗种植体周粘膜炎患者、③并随机选取30例成功病例30颗健康种植体作对照组。
2．临床检查：记录所有研究对象的种植体探诊深度并拍摄全口曲面体层片。
3．龈下菌斑的采集及处理：由同一名医师从每个种植体的颊(唇)侧的近中至远中用无菌碳纤维刮治器收集龈下菌斑，标本立即置入1ml PBS溶液中、-20℃冻存。
4．龈下菌斑DNA的提取。
5．聚合酶链反应(PCR)检测标本中的微生物：检测临床标本中的7种牙周可疑致病微生物：伴放线放线杆菌(A.a)、中间普氏菌(P.i)、牙龈卟啉单胞菌(P.g)、齿垢密螺旋体(T.d)、变黑普氏菌(P.n)、直肠弯曲菌(C.r)、福赛拟杆菌(B.f)的分布情况。
结果：
1．三组中各种细菌的检出率各不相同
(1)种植体周粘膜炎组T.d、P.i和B.f的检出率高于健康组，种植体周围炎组P.g、B.f、T.d、P.n的检出率高于健康组，种植体周围炎组的P.g、T.d、P.n高于种植体周粘膜炎组，差异均有统计学意义(P＜0.05)。
(2)A.a和C.r的检出率在3组间差异无统计学意义(P＞0.05)。
2．进行logistic逐步回归分析，发现P.g、T.d和P.n三种细菌对种植体周围炎的影响较为显著(OR值分别为5.830、61.037和3.672，均大于1)。
结论：
1．与种植体周围软硬组织健康相关的微生物，主要集中在P.g、P.i、P.n、B.f、T.d，这些菌种的协同作用很可能就是种植体周围炎的致病因素；当P.g、T.d和P.n共同出现时对种植体周围骨吸收影响较为显著；P.i、T.d、B.f增多时，种植体龈沟有加深趋势；种植体周围炎可能是引起种植体周粘膜炎的致病菌进一步繁殖增生造成。
3．采用聚合酶链反应扩增16SrRNA基因(16SrDNA)片段的方法，可以准确地检测种植体周特定的可疑致病微生物，这种方式有望早期发现种植体周围炎，有助于及时防治种植体周围炎的发生。
Objective:
1. To detect the colonization of 7 subgingival suspected pathogens in partialedentulous patients with and without peri-implantitis.
2. To investigate the relationship of periodontal suspected pathogens, peri-implantitisand the bone loss.
3. To investigate the microbiologic pathogeny of peri-implantitis, and to find the mainpathogen of peri-implantitis in the early stage.
4. To find a method to diagnosis peri-implantitis with early prevention and treatment.
Methods:
1. 88 patients with 89 implants were divided into 3 groups: 1) peri-implantitis group,29 peri-implantitis-patients with 29 implants 2) peri-mucositis group, 29peri-mucositis-patients with 30 implants 3) healthy group, 30 healthy patients with30 implants by random.
2. Peri-implant pocket depth (PPD) and oral pan tomography was recorded as clinicalparameters.
3. Buccal subgingival plaque was collected from mesial to distal by same dentist withnon-bacterial carbonous fibred tool.
4. DNA of subgingival plaque was distilled.
5. Colonization of 7 periodontal suspected pathogens was detected by polymerasechain reaction (PCR): A. actinomyceremcomitans (A. a)、P. intermedia (P. i)、P.gingivalis(P. g)、Treponema denticola (T. d)、P nigrescens(P. n)、Campylobacterrectus (C. r)、B. forsythus (B. f).
Results:
1. Prevalence of these 3 groups was different.
(1) Higher detection of B.f、P.i and T.d was found in peri-mucositis group than in healthy group. Higher detection of P.g、B.f、T.d and P.n was found inperi-implantitis group than in healthy group. Higher detection of P.n、P.g and T.dwas found in peri-implantitis group than in peri-mucositis group. There issignificant difference among these 3 groups (P＜0.05).
(2) No significant difference existed among these 3 groups in detections of A.a andC.r (P＞0.05).
2. Logistic analyze: P.g、T.d and P.n played an important role in peri-implantitis. (OR=5.830, 61.037 and3.672).
Conclusion:
1. P.g、P.i、P.n、B.f and T.d are major bacterium that relate with soft and hardtissue around the implant, especially with peri-implantitis. P.g cooperated with P.nand T.d, might cause bone loss around implant. When the detection of P.i、T.dand B.f increase, peri-implant pocket depth has deepened trend; The furtherreproduction of pathogens of peri-mucositis may be the reason for peri-implantitis.
2. Suspected pathogens of peri-implantitis were detected by using 16SrRNA basedPCR, which could be helpful with early diagnosis, prevention and cure ofperi-implantitis.
1．检测健康及炎症种植体龈下菌斑中7种牙周可疑致病微生物的定殖情况；
2．探讨牙周可疑致病微生物与种植体周围炎症及骨吸收的关系；
3．探讨种植体周病变的微生物学病因，寻找能早期反映种植体周病变的主要致病微生物；
4．探索早期诊断及早期防治种植体周围炎的方法。
方法：
1．病例收集分组：选择种植修复后12个月以上的88位患者的89颗种植体，分三组①种植体周围炎组29例29颗种植体周围炎患者、②种植体周粘膜炎组29例30颗种植体周粘膜炎患者、③并随机选取30例成功病例30颗健康种植体作对照组。
2．临床检查：记录所有研究对象的种植体探诊深度并拍摄全口曲面体层片。
3．龈下菌斑的采集及处理：由同一名医师从每个种植体的颊(唇)侧的近中至远中用无菌碳纤维刮治器收集龈下菌斑，标本立即置入1ml PBS溶液中、-20℃冻存。
4．龈下菌斑DNA的提取。
5．聚合酶链反应(PCR)检测标本中的微生物：检测临床标本中的7种牙周可疑致病微生物：伴放线放线杆菌(A.a)、中间普氏菌(P.i)、牙龈卟啉单胞菌(P.g)、齿垢密螺旋体(T.d)、变黑普氏菌(P.n)、直肠弯曲菌(C.r)、福赛拟杆菌(B.f)的分布情况。
结果：
1．三组中各种细菌的检出率各不相同
(1)种植体周粘膜炎组T.d、P.i和B.f的检出率高于健康组，种植体周围炎组P.g、B.f、T.d、P.n的检出率高于健康组，种植体周围炎组的P.g、T.d、P.n高于种植体周粘膜炎组，差异均有统计学意义(P＜0.05)。
(2)A.a和C.r的检出率在3组间差异无统计学意义(P＞0.05)。
2．进行logistic逐步回归分析，发现P.g、T.d和P.n三种细菌对种植体周围炎的影响较为显著(OR值分别为5.830、61.037和3.672，均大于1)。
结论：
1．与种植体周围软硬组织健康相关的微生物，主要集中在P.g、P.i、P.n、B.f、T.d，这些菌种的协同作用很可能就是种植体周围炎的致病因素；当P.g、T.d和P.n共同出现时对种植体周围骨吸收影响较为显著；P.i、T.d、B.f增多时，种植体龈沟有加深趋势；种植体周围炎可能是引起种植体周粘膜炎的致病菌进一步繁殖增生造成。
3．采用聚合酶链反应扩增16SrRNA基因(16SrDNA)片段的方法，可以准确地检测种植体周特定的可疑致病微生物，这种方式有望早期发现种植体周围炎，有助于及时防治种植体周围炎的发生。
Objective:
1. To detect the colonization of 7 subgingival suspected pathogens in partialedentulous patients with and without peri-implantitis.
2. To investigate the relationship of periodontal suspected pathogens, peri-implantitisand the bone loss.
3. To investigate the microbiologic pathogeny of peri-implantitis, and to find the mainpathogen of peri-implantitis in the early stage.
4. To find a method to diagnosis peri-implantitis with early prevention and treatment.
Methods:
1. 88 patients with 89 implants were divided into 3 groups: 1) peri-implantitis group,29 peri-implantitis-patients with 29 implants 2) peri-mucositis group, 29peri-mucositis-patients with 30 implants 3) healthy group, 30 healthy patients with30 implants by random.
2. Peri-implant pocket depth (PPD) and oral pan tomography was recorded as clinicalparameters.
3. Buccal subgingival plaque was collected from mesial to distal by same dentist withnon-bacterial carbonous fibred tool.
4. DNA of subgingival plaque was distilled.
5. Colonization of 7 periodontal suspected pathogens was detected by polymerasechain reaction (PCR): A. actinomyceremcomitans (A. a)、P. intermedia (P. i)、P.gingivalis(P. g)、Treponema denticola (T. d)、P nigrescens(P. n)、Campylobacterrectus (C. r)、B. forsythus (B. f).
Results:
1. Prevalence of these 3 groups was different.
(1) Higher detection of B.f、P.i and T.d was found in peri-mucositis group than in healthy group. Higher detection of P.g、B.f、T.d and P.n was found inperi-implantitis group than in healthy group. Higher detection of P.n、P.g and T.dwas found in peri-implantitis group than in peri-mucositis group. There issignificant difference among these 3 groups (P＜0.05).
(2) No significant difference existed among these 3 groups in detections of A.a andC.r (P＞0.05).
2. Logistic analyze: P.g、T.d and P.n played an important role in peri-implantitis. (OR=5.830, 61.037 and3.672).
Conclusion:
1. P.g、P.i、P.n、B.f and T.d are major bacterium that relate with soft and hardtissue around the implant, especially with peri-implantitis. P.g cooperated with P.nand T.d, might cause bone loss around implant. When the detection of P.i、T.dand B.f increase, peri-implant pocket depth has deepened trend; The furtherreproduction of pathogens of peri-mucositis may be the reason for peri-implantitis.
2. Suspected pathogens of peri-implantitis were detected by using 16SrRNA basedPCR, which could be helpful with early diagnosis, prevention and cure ofperi-implantitis.
引文
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2．宿玉成．现代口腔种植学[M]．北京：人民卫生出版社，2004：383．
3. Hanisch O, Cortella CA, Boskovic MM, et al. Experimental peri-implant tissue breakdown around hydroxyapatite-coated implants [J]. J Periodontol, 1997, 68 (1): 59—66.
4. Augthun M, Conrads G. Microbial findings of deep peri-implant bone defects [J]. J Oral Maxillofac Implants, 1997, 12(1): 106-112.
5. Adell R, Eriksson B, Lekholm U, et al. Long term follow-up study of Osseo integrated implants in the treatment of totally edentulous jaws [J]. J Oral Maxillofac Implants, 1990, 5: 347-359.
6. Higuchi KW, Folmer T, Christina. Implant survival rates in partially edentulous patients: a 3-year prospective multicenter study [J]. J Oral Maxillofac Surg, 1995, 53: 264-268.
7. Schou S, llolmstrup P, Kornman K. Ligature-induced marginal inflammation around osseointegrated implants and ankylosed teeth: stereologic and histologic observations in cynomolgus monkeys [J]. J Periodontol, 1993, 64: 529-537.
8. Berglundh T, Lindhe J, Ericsson I et al. The soft tissue barrier at implants and teeth [J]. J Clin Oral Impl Res, 1991, 2: 81-90.
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