基础治疗对重度牙周炎患者龈沟液内中性粒细胞胞外诱捕网形成的影响研究

doi:10.7518/hxkq.2024.2024162

华西口腔医学杂志 ›› 2025, Vol. 43 ›› Issue (1): 46-52.doi: 10.7518/hxkq.2024.2024162

基础治疗对重度牙周炎患者龈沟液内中性粒细胞胞外诱捕网形成的影响研究

付岚清(), 郝新宇, 钱文博, 孙颖()

南京医科大学附属口腔医院牙周病科口腔疾病研究与防治国家级重点实验室培育建设点江苏省口腔转化医学工程研究中心，南京 210029

收稿日期:2024-04-24 修回日期:2024-08-06 出版日期:2025-02-01 发布日期:2025-01-22
通讯作者: 孙颖 E-mail:2577334820@qq.com;ebolasun@njmu.edu.cn
作者简介:付岚清，住院医师，硕士，E-mail：2577334820@qq.com
基金资助:
江苏省自然科学基金(BK20231268);江苏省科教能力提升工程——江苏省研究型医院(YJXYYJSDW4);江苏省医学创新中心(CXZX202227)

Effects of initial periodontal therapy on the formation of neutrophil extracellular traps in gingival crevicular fluid in patients with severe periodontitis

Fu Lanqing(), Hao Xinyu, Qian Wenbo, Sun Ying()

Dept. of Periodontology, The Affiliated Stomatological Hospital of Nanjing Medical University & State Key Laboratory Cultivation Base of Research, Prevention and Treatment for Oral Diseases & Jiangsu Province Engineering Research Center of Stomatological Translational Medicine, Nanjing 210029, China

Received:2024-04-24 Revised:2024-08-06 Online:2025-02-01 Published:2025-01-22
Contact: Sun Ying E-mail:2577334820@qq.com;ebolasun@njmu.edu.cn
Supported by:
Natural Science Foundation of Jiangsu Province(BK20231268);Jiangsu Province Capability Improvement Project through Science, Technology and Education, Jiangsu Provincial Research Hospital Cultivation Unit(YJX-YYJSDW4);Jiangsu Provincial Medical Innovation Center(CXZX202227);Correspondence: Sun Ying, E-mail: ebolasun@njmu.edu.cn

摘要/Abstract

摘要：

目的探讨基础治疗对重度牙周炎患者龈沟液（GCF）内中性粒细胞胞外诱捕网（NETs）水平的影响，并分析影响NETs形成的因素。方法纳入Ⅲ~Ⅳ期牙周炎患者31例，初诊及基础治疗后6~8周，记录牙周临床指标菌斑指数（PLI）、牙龈指数（GI）、探诊深度（PD）和临床附着丧失（CAL），采用免疫荧光技术检测GCF内NETs水平，实时荧光定量聚合酶链式反应（qPCR）检测龈下非附着菌斑中总菌、牙龈卟啉单胞菌（P. gingivalis）、伴放线菌团聚杆菌（A. actinomycetemcomitans）和中间普雷沃菌（P. intermedia）数量，酶联免疫吸附试验检测GCF中肿瘤坏死因子-α（TNF-α）和白细胞介素-8（IL-8）水平，并分析NETs水平与上述指标的相关性。结果基础治疗后，GCF内NETs水平，PLI、GI、PD、CAL等牙周临床指标，总菌、P. gingivalis、A. actinomycetemcomitans及P. intermedia数量，IL-8及TNF-α水平均显著下降（P<0.05）。GCF内NETs水平与PLI、GI、PD、CAL、TNF-α、IL-8水平，总菌和P. gingivalis数量存在高度正相关性（P<0.05）。结论基础治疗能降低重度牙周炎患者GCF中的NETs水平，后者的形成可能与P. gingivalis数量和GCF中TNF-α、IL-8水平相关。

关键词: 牙周炎, 基础治疗, 龈沟液, 中性粒细胞胞外诱捕网

Abstract:

Objective This study aimed to observe the effects of initial periodontal therapy on the level of neutrophil extracellular traps (NETs) in gingival crevicular fluid (GCF) of patients with severe periodontitis and to analyze the factors related to the formation of NETs. Methods Thirty-one patients with stage Ⅲ-Ⅳ periodontitis were recruited. Clinical periodontal parameters, including plaque index (PLI), gingival index (GI), probing depth (PD), and clinical atta-chment loss (CAL), were recorded before and 6-8 weeks after initial periodontal therapy. Levels of NETs in GCF were detected by immunofluorescence staining. Quantities of total bacteria, Porphyromonas gingivalis (P. gingivalis), Aggregatibacter actinomycetemcomitans (A. actionomycetemcomitans) and Prevotella intermedia (P. intermedia)in unattached subgingival plaque were determined by real-time quantitative PCR, and levels of tumor necrosis factor-α (TNF-α) and interleukin-8 (IL-8) in GCF were explored by enzyme-linked immunosorbent assay. In addition, the correlations between the level of NETs and the above indicators were analyzed. Results After initial periodontal therapy, the level of NETs in GCF, PLI, GI, PD, and CAL; quantities of total bacteria, P. gingivalis, A. actinomycetemcomitans, and P. itermedia; and levels of IL-8 and TNF-α significantly decreased (P<0.05). We observed strong positive correlations between the level of NETs and PLI, GI, PD, CAL, the amount of total bacteria, P. gingivalis, TNF-α, and IL-8 (P<0.05). Conclusion Initial periodontal therapy might decrease the level of NETs in GCF from patients with severe periodontitis, which might be positively correlated with the quantities of P. gingivalis andthe levels of TNF-α and IL-8 in GCF.

Key words: periodontitis, initial periodontal therapy, gingival crevicular fluid, neutrophil extracellular trap

中图分类号:

R781.4

付岚清, 郝新宇, 钱文博, 孙颖. 基础治疗对重度牙周炎患者龈沟液内中性粒细胞胞外诱捕网形成的影响研究[J]. 华西口腔医学杂志, 2025, 43(1): 46-52.

Fu Lanqing, Hao Xinyu, Qian Wenbo, Sun Ying. Effects of initial periodontal therapy on the formation of neutrophil extracellular traps in gingival crevicular fluid in patients with severe periodontitis[J]. West China Journal of Stomatology, 2025, 43(1): 46-52.

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参考文献 27

1	孟焕新. 2018年牙周病和植体周病国际新分类简介[J]. 中华口腔医学杂志, 2019, 54(2): 73-78.
	Meng HX. 2018 world new classification of periodontal and peri-implant diseases and conditions[J]. Chin J Stomatol, 2019, 54(2): 73-78.
2	Hajishengallis G, Chavakis T. Local and systemic mechanisms linking periodontal disease and inflammatory comorbidities[J]. Nat Rev Immunol, 2021, 21(7): 426-440.
3	Brinkmann V, Reichard U, Goosmann C, et al. Neutrophil extracellular traps kill bacteria[J]. Science, 2004, 303(5663): 1532-1535.
4	Jiang QS, Zhao YX, Shui YS, et al. Interactions between neutrophils and periodontal pathogens in late-onset pe-riodontitis[J]. Front Cell Infect Microbiol, 2021, 11: 627328.
5	Vitkov L, Klappacher M, Hannig M, et al. Neutrophil fate in gingival crevicular fluid[J]. Ultrastruct Pathol, 2010, 34(1): 25-30.
6	Magán-Fernández A, O’Valle F, Abadía-Molina F, et al. Characterization and comparison of neutrophil extracellular traps in gingival samples of periodontitis and gingivitis: a pilot study[J]. J Periodontal Res, 2019, 54(3): 218-224.
7	Wigerblad G, Kaplan MJ. Neutrophil extracellular traps in systemic autoimmune and autoinflammatory diseases[J]. Nat Rev Immunol, 2023, 23(5): 274-288.
8	Vitkov L, Klappacher M, Hannig M, et al. Extracellular neutrophil traps in periodontitis[J]. J Periodontal Res, 2009, 44(5): 664-672.
9	Masuda S, Nakazawa D, Shida H, et al. NETosis markers: quest for specific, objective, and quantitative markers[J]. Clin Chim Acta, 2016, 459: 89-93.
10	Al-Bakri SMR, Magan-Fernandez A, Galindo-Moreno P, et al. Detection and comparison of neutrophil extracellular traps in tissue samples of peri-implantitis, periodontitis, and healthy patients: a pilot study[J]. Clin Implant Dent Relat Res, 2024, 26(3): 631-641.
11	Goggs R, Jeffery U, LeVine DN, et al. Neutrophil-extracellular traps, cell-free DNA, and immunothrombosis in companion animals: a review[J]. Vet Pathol, 2020, 57(1): 6-23.
12	Peng Y, Wu X, Zhang S, et al. The potential roles of type Ⅰinterferon activated neutrophils and neutrophil extracellular traps (NETs) in the pathogenesis of primary Sjögren’s syndrome[J]. Arthritis Res Ther, 2022, 24(1): 170.
13	Arends EJ, Dam LSV, Kraaij T, et al. A high-throughput assay to assess and quantify neutrophil extracellular trap formation[J]. J Vis Exp, 2019, 143. doi: 10.3791/59150 .
14	Ghazizadeh M, Sasaki Y, Oguro T, et al. A novel technique for observing the internal ultrastructure of human chromosomes with known karyotype[J]. Microsc Microanal, 2008, 14(4): 357-361.
15	Bryzek D, Ciaston I, Dobosz E, et al. Triggering NETosis via protease-activated receptor (PAR)-2 signaling as a mechanism of hijacking neutrophils function for pathogen benefits[J]. PLoS Pathog, 2019, 15(5): e1007773.
16	Hirschfeld J, Roberts HM, Chapple IL, et al. Effects of Aggregatibacter actinomycetemcomitans leukotoxin on neutrophil migration and extracellular trap formation[J]. J Oral Microbiol, 2016, 8: 33070.
17	Baz AA, Hao HF, Lan SM, et al. Neutrophil extracellular traps in bacterial infections and evasion strategies[J]. Front Immunol, 2024, 15: 1357967.
18	Gonçalves PF, Klepac-Ceraj V, Huang H, et al. Correlation of Aggregatibacter actinomycetemcomitans detection with clinical/immunoinflammatory profile of localized aggressive periodontitis using a 16S rRNA microarray method: a cross-sectional study[J]. PLoS One, 2013, 8(12): e85066.
19	He JY, Huang WJ, Pan ZW, et al. Quantitative analysis of microbiota in saliva, supragingival, and subgingival plaque of Chinese adults with chronic periodontitis[J]. Clin Oral Investig, 2012, 16(6): 1579-1588.
20	Tan KS, Woo CH, Ong G, et al. Prevalence of Actinobacillus actinomycetemcomitans in an ethnic adult Chinese population[J]. J Clin Periodontol, 2001, 28(9): 886-890.
21	Liu GJ, Luan QX, Chen F, et al. Shift in the subgingival microbiome following scaling and root planing in gene-ralized aggressive periodontitis[J]. J Clin Periodontol, 2018, 45(4): 440-452.
22	Kovalčíková AG, Novák B, Roshko O, et al. Extracellular DNA and markers of neutrophil extracellular traps in saliva from patients with periodontitis-a case-control study[J]. J Clin Med, 2024, 13(2): 468.
23	Moonen CG, Buurma KG, Faruque MR, et al. Periodontal therapy increases neutrophil extracellular trap degradation[J]. Innate Immun, 2020, 26(5): 331-340.
24	White P, Sakellari D, Roberts H, et al. Peripheral blood neutrophil extracellular trap production and degradation in chronic periodontitis[J]. J Clin Periodontol, 2016, 43(12): 1041-1049.
25	Moonen CGJ, Hirschfeld J, Cheng LL, et al. Oral neu-trophils characterized: chemotactic, phagocytic, and neutrophil extracellular trap (NET) formation properties[J]. Front Immunol, 2019, 10: 635.
26	Czerwińska J, Kasprowicz-Furmańczyk M, Placek W, et al. Changes in tumor necrosis factor α (TNFα) and peptidyl arginine deiminase 4 (PAD-4) levels in serum of general treated psoriatic patients[J]. Int J Environ Res Public Health, 2022, 19(14): 8723.
27	Shu QX, Zhang N, Liu YY, et al. IL-8 triggers neutrophil extracellular trap formation through an nicotinami-de adenine dinucleotide phosphate oxidase- and mitogen-activated protein kinase pathway-dependent mechanism in uveitis[J]. Invest Ophthalmol Vis Sci, 2023, 64(13): 19.

时间	PLI	GI	PD/mm	CAL/mm
治疗前	2..46±0.25	2.53±0.26	7.33±0.18	5.67±0.22
治疗后	1.17±0.21*	1.18±0.24*	5.02±0.19*	3.15±0.17*

指标	r	P值
PLI	0.608	0.012*
GI	0.822	0.023*
PD	0.886	0.019*
CAL	0.893	0.007**
TNF-α	0.833	0.01*
IL-8	0.800	0.009**
总菌	0.748	0.005**
P. gingivalis	0.718	0.006**
A. actinomycetemcomitans	0.619	0.102
P. intermedia	0.143	0.76

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基础治疗对重度牙周炎患者龈沟液内中性粒细胞胞外诱捕网形成的影响研究

Effects of initial periodontal therapy on the formation of neutrophil extracellular traps in gingival crevicular fluid in patients with severe periodontitis

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