Int J Stomatol ›› 2024, Vol. 51 ›› Issue (3): 288-295.doi: 10.7518/gjkq.2024044

• Original Articles • Previous Articles     Next Articles

Correlation study of plasma cell-free extra-mitochondrial mitochondria DNA and periodontitis clinical parameters

Zaimu Yang1(),Pei Cao1,Zhenhua Liu2,Qingxian Luan1()   

  1. 1.Dept. of Periodontology, Peking University School and Hospital of Stomatology & National Center for Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Laboratory for Digital and Material Technology of Stomatology & Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China
    2.Dept. of Stomatology, Beijing Zhongguancun Hospital, Beijing 100190, China
  • Received:2023-08-16 Revised:2024-02-01 Online:2024-05-01 Published:2024-05-06
  • Contact: Qingxian Luan;


Objective Plasma cell-free extra-cellular mitochondrial DNA (cf-exmtDNA) shows pro-inflammatory potential. This study aims to investigate the correlation between cf-exmtDNA and baseline periodontitis clinical parameters in patients in general health. Methods A total of 78 participants aged 18-45 years were enrolled: 11 periodontal healthy volunteers, 11 patients with gingivitis, and 56 patients with periodontitis. Baseline periodontal index, age, gender, body mass index (BMI), and baseline fasting blood glucose (FBG) were examined and recorded. Anticoagulated venous blood (4 mL) was collected, and cf-mtDNA was extracted by secondary centrifugation. cf-exmtDNA concentration was measured through real-time quantitative polymerase chain reaction. Different states of periodontal inflammation were compared in terms of plasma cf-exmtDNA copy number, and the relationship among plasma cf-exmtDNA, mean probing depth (mPD), mean clinical attachment level (mCAL), mean bleeding index (mBI), mean plaque index, age, FBG, and BMI were examined through correlation and multiple linear regression analyses. Results Patients with periodontitis had significantly higher plasma cf-exmtDNA levels than the healthy volunteers (P=0.042). The overall plasma cf-exmtDNA was significantly positively correlated with age (P=0.023), mPD (P<0.001), mCAL (P=0.006), and mBI (P=0.026). Multiple regression analysis showed that plasma cf-exmtDNA level was significantly dependent on mPD. Conclusion In the general population aged 18–45, patients with periodontitis had significantly higher plasma cf-exmt-DNA level than the healthy volunteers, and plasma cf-exmtDNA concentration was significantly positively correlated with age, mPD, mCAL, and mBI.

Key words: periodontitis, cell-free DNA, mitochondrial DNA, cross-sectional study, periodontal clinical parameters

CLC Number: 

  • R781.4


Tab 1

qPCR program setting"

保持期502 min
预变性9510 min
PCR期(40 个循环)变性9510 s
退火6015 s
延伸a7230 s

Tab 2

Primer sequences"


Tab 3

Baseline characteristics of periodontal healthy, gingivitis, periodontitis participants n=78"


Fig 1

The relationship between plasma cf-exmtDNA and periodontal inflammation status and clinical indicators"

Tab 4

Correlations between clinical parameters and plasma cf-exmtDNA"

BMI /(kg/m20.2060.071

Tab 5

Multiple linear regression of plasma cf-exmtDNA and periodontal clinical parameters"

多重线性回归模型模型参数B95% CIβtP
血浆cf-exmtDNA常量-1 257.091(-2 090.559,-423.622)-3.0040.004

Fig 2

Multiple variables bubble plot (left) and multiple linear regression model (right) of mPD"

1 Hajishengallis G, Chavakis T, Lambris JD. Current understanding of periodontal disease pathogenesis and targets for host-modulation therapy[J]. Perio-dontol 2000, 2020, 84(1): 14-34.
2 Bae JH, Jo SI, Kim SJ, et al. Circulating cell-free mtDNA contributes to AIM2 inflammasome-mediated chronic inflammation in patients with type 2 diabetes[J]. Cells, 2019, 8(4): 328.
3 Duvvuri B, Lood C. Cell-free DNA as a biomarker in autoimmune rheumatic diseases[J]. Front Immunol, 2019, 10: 502.
4 Aswani A, Manson J, Itagaki K, et al. Scavenging circulating mitochondrial DNA as a potential therapeutic option for multiple organ dysfunction in trauma hemorrhage[J]. Front Immunol, 2018, 9: 891.
5 Zhang Q, Raoof M, Chen Y, et al. Circulating mitochondrial DAMPs cause inflammatory responses to injury[J]. Nature, 2010, 464(7285): 104-107.
6 Nasi M, Bianchini E, de Biasi S, et al. Increased plasma levels of mitochondrial DNA and pro-inflammatory cytokines in patients with progressive multiple sclerosis[J]. J Neuroimmunol, 2020, 338: 577107.
7 Singel KL, Grzankowski KS, ANMNHKhan, et al. Mitochondrial DNA in the tumour microenvironment activates neutrophils and is associated with worse outcomes in patients with advanced epithelial ovarian cancer[J]. Br J Cancer, 2019, 120(2): 207-217.
8 Ward GA, McGraw KL, Abbas-Aghababazadeh F, et al. Oxidized mitochondrial DNA released after inflammasome activation is a disease biomarker for myelodysplastic syndromes[J]. Blood Adv, 2021, 5(8): 2216-2228.
9 Pinti M, Cevenini E, Nasi M, et al. Circulating mitochondrial DNA increases with age and is a familiar trait: Implications for “inflamm-aging”[J]. Eur J Immunol, 2014, 44(5): 1552-1562.
10 Trumpff C, Marsland AL, Basualto-Alarcón C, et al. Acute psychological stress increases serum circulating cell-free mitochondrial DNA[J]. Psychoneuroendocrinology, 2019, 106: 268-276.
11 Trumpff C, Michelson J, Lagranha CJ, et al. Stress and circulating cell-free mitochondrial DNA: a systematic review of human studies, physiological considerations, and technical recommendations[J]. Mitochondrion, 2021, 59: 225-245.
12 Trumpff C, Rausser S, Haahr R, et al. Dynamic behavior of cell-free mitochondrial DNA in human saliva[J]. Psychoneuroendocrinology, 2022, 143: 105852.
13 Al Amir Dache Z, Otandault A, Tanos R, et al. Blood contains circulating cell-free respiratory competent mitochondria[J]. FASEB J, 2020, 34(3): 3616-3630.
14 Roch B, Pisareva E, Sanchez C, et al. Plasma derived cell-free mitochondrial DNA originates mainly from circulating cell-free mitochondria[J]. bio Rxiv, 2021. doi: 10.1101/2021.09.03.458846 .
doi: 10.1101/2021.09.03.458846
15 Stephens OR, Grant D, Frimel M, et al. Characteri-zation and origins of cell-free mitochondria in healthy murine and human blood[J]. Mitochondrion, 2020, 54: 102-112.
16 Lázaro-Ibáñez E, Lässer C, Shelke GV, et al. DNA analysis of low- and high-density fractions defines heterogeneous subpopulations of small extracellular vesicles based on their DNA cargo and topology[J]. J Extracell Vesicles, 2019, 8(1): 1656993.
17 Szilágyi M, Pös O, Márton É, et al. Circulating cell-free nucleic acids: Main characteristics and clinical application[J]. Int J Mol Sci, 2020, 21(18): 6827.
18 Pérez-Treviño P, Velásquez M, García N. Mechanisms of mitochondrial DNA escape and its relationship with different metabolic diseases[J]. Biochim Biophys Acta Mol Basis Dis, 2020, 1866(6): 165761.
19 Liu DL, Gao YS, Liu J, et al. Intercellular mitochondrial transfer as a means of tissue revitalization[J]. Signal Transduct Target Ther, 2021, 6(1): 65.
20 Tsilioni I, Theoharides TC. Extracellular vesicles are increased in the serum of children with autism spectrum disorder, contain mitochondrial DNA, and stimulate human microglia to secrete IL-1β[J]. J Neuroinflammation, 2018, 15(1): 239.
21 de Gaetano A, Solodka K, Zanini G, et al. Molecular mechanisms of mtDNA-mediated inflammation[J]. Cells, 2021, 10(11): 2898.
22 Liu J, Wang YF, Shi Q, et al. Mitochondrial DNA efflux maintained in gingival fibroblasts of patients with periodontitis through ROS/mPTP pathway[J]. Oxid Med Cell Longev, 2022, 2022: 1000213.
23 Masi S, Orlandi M, Parkar M, et al. Mitochondrial oxidative stress, endothelial function and metabolic control in patients with type Ⅱ diabetes and perio-dontitis: a randomised controlled clinical trial[J]. Int J Cardiol, 2018, 271: 263-268.
24 Sun XY, Mao YX, Dai PP, et al. Mitochondrial dysfunction is involved in the aggravation of periodontitis by diabetes[J]. J Clin Periodontol, 2017, 44(5): 463-471.
25 Konečná B, Gaál Kovalčíková A, Pančíková A, et al. Salivary extracellular DNA and DNase activity in periodontitis[J]. Appl Sci, 2020, 10(21): 7490.
26 吴圣贤, 王成祥. 临床研究样本含量估算基础[M]. 北京: 人民卫生出版社, 2008: 24-26.
Wu SX, Wang CX. Sample size calculation basics for clinical research[M]. Beijing: People’s Medical Publishing House, 2008: 24-26.
27 Papapanou PN, Sanz M, Buduneli N, et al. Perio-dontitis: consensus report of workgroup 2 of the 2017 World Workshop on the Classification of Perio-dontal and Peri-Implant Diseases and Conditions[J]. J Periodontol, 2018, 89(): S173-S182.
28 Lang NP, Bartold PM. Periodontal health[J]. J Perio-dontol, 2018, 89(): S9-S16.
29 Hummel EM, Hessas E, Müller S, et al. Cell-free DNA release under psychosocial and physical stress conditions[J]. Transl Psychiatry, 2018, 8(1): 236.
30 Bustin SA, Benes V, Garson JA, et al. The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments[J]. Clin Chem, 2009, 55(4): 611-622.
31 Lumley T, Diehr P, Emerson S, et al. The importance of the normality assumption in large public health data sets[J]. Annu Rev Public Health, 2002, 23: 151-169.
32 Brinkmann V. Neutrophil extracellular traps in the second decade[J]. J Innate Immun, 2018, 10(5/6): 414-421.
33 Cataño Cañizales YG, Uresti Rivera EE, García Jacobo RE, et al. Increased levels of AIM2 and circulating mitochondrial DNA in type 2 diabetes[J]. Iran J Immunol, 2018, 15(2): 142-155.
34 Silzer T, Barber R, Sun J, et al. Circulating mitochondrial DNA: new indices of type 2 diabetes-related cognitive impairment in Mexican Americans[J]. PLoS One, 2019, 14(3): e0213527.
35 Fatima T, Khurshid Z, Rehman A, et al. Gingival crevicular fluid (GCF): a diagnostic tool for the detection of periodontal health and diseases[J]. Molecules, 2021, 26(5): 1208.
[1] Yuhong Ma,Lei Zhao. Process and progress in the clinical research of minimally invasive non-operative periodontal therapy technology [J]. Int J Stomatol, 2024, 51(2): 227-232.
[2] Fu Yu, He Wei, Huang Lan. Ferroptosis and its implication in oral diseases [J]. Int J Stomatol, 2024, 51(1): 36-44.
[3] Luo Xiaojie,Wang Dexu,Chen Xiaotao. Relationship between periodontitis and ferroptosis based on bioinformatics analysis [J]. Int J Stomatol, 2023, 50(6): 661-668.
[4] Huang Yuanhong,Peng Xian,Zhou Xuedong.. Progress in research into the effect of Rhizoma Drynariae on the treatment of bone-related diseases in the oral cavity [J]. Int J Stomatol, 2023, 50(6): 679-685.
[5] Hu Jia,Wang Xiuqing,Lu Guoying,Huang Xiaojing.. Regenerative endodontic procedures for permanent tooth with immature apices in adult patients [J]. Int J Stomatol, 2023, 50(6): 686-695.
[6] Gong Meiling,Cheng Xingqun,Wu Hongkun.. Research progress on the correlation between Parkinson’s disease and periodontitis [J]. Int J Stomatol, 2023, 50(5): 587-593.
[7] Xu Zhibo,Meng Xiuping.. Research progress on mechanism of Enterococcus faecalis escaping host immune defense [J]. Int J Stomatol, 2023, 50(5): 613-617.
[8] Sun Jia,Han Ye,Hou Jianxia. Research progress on the role of interleukin-6-hepcidin signal axis in regulating the pathogenesis of periodontitis-associated anemia [J]. Int J Stomatol, 2023, 50(3): 329-334.
[9] Liang Zhiying,Zhao Yuanxi,Zhu Jiani,Su Qin.. Retrospective analysis of clinical data of 288 cases of endodontic microsurgery on anterior teeth [J]. Int J Stomatol, 2023, 50(2): 166-171.
[10] Liu Tiqian,Liang Xing,Liu Weiqing,Li Xiaohong,Zhu Rui.. Research progress on the role and mechanism of occlusal trauma in the development of periodontitis [J]. Int J Stomatol, 2023, 50(1): 19-24.
[11] Li Qiong,Yu Weixian. Research progress on resveratrol for the treatment of periodontitis and its bioavailability [J]. Int J Stomatol, 2023, 50(1): 25-31.
[12] Huang Weikun,Xu Qiuyan,Zhou Ting.. Role of baicalin and mechanisms through which baicalin attenuates oxidative stress injury induced by lipopolysaccharide on macrophages [J]. Int J Stomatol, 2022, 49(5): 521-528.
[13] Zhou Jianpeng,Xie Xudong,Zhao Lei,Wang Jun.. Research progress on the roles and mechanisms of T-helper 17 cells and interleukin-17 in periodontitis [J]. Int J Stomatol, 2022, 49(5): 586-592.
[14] Chen Huiyu,Bai Mingru,Ye Ling.. Progress in understanding the correlations between semaphorin 3A and common oral diseases [J]. Int J Stomatol, 2022, 49(5): 593-599.
[15] Zhou Jiajia,Zhao Lei,Xu Xin. Research progress on the genetic polymorphism of periodontitis [J]. Int J Stomatol, 2022, 49(4): 432-440.
Full text



[1] . [J]. Foreign Med Sci: Stomatol, 1999, 26(06): .
[2] . [J]. Foreign Med Sci: Stomatol, 1999, 26(06): .
[3] . [J]. Foreign Med Sci: Stomatol, 1999, 26(06): .
[4] . [J]. Foreign Med Sci: Stomatol, 1999, 26(06): .
[5] . [J]. Foreign Med Sci: Stomatol, 1999, 26(06): .
[6] . [J]. Foreign Med Sci: Stomatol, 1999, 26(05): .
[7] . [J]. Foreign Med Sci: Stomatol, 1999, 26(04): .
[8] . [J]. Foreign Med Sci: Stomatol, 1999, 26(04): .
[9] . [J]. Foreign Med Sci: Stomatol, 2004, 31(02): 146 -148 .
[10] . [J]. Foreign Med Sci: Stomatol, 2005, 32(06): 461 -462 .