Correlation between cell free DNA in gingival crevicular fluid and clinical periodontal parameters by using two collection techniques
Article Sidebar
Main Article Content
Abstract
Objectives: The purpose of this study was to evaluate the correlation between cell free DNA (cfDNA) in gingival crevicular fluid (GCF) and clinical periodontal parameters and to compare the efficiency of the two GCF collection techniques.
Materials and Methods: Clinical periodontal parameters including pocket depth, clinical attachment level and bleeding on probing were recorded in forty teeth of twenty patients with moderate to severe periodontal disease. GCF samples were collected from each tooth by two collection techniques. First, it was randomly collected by either modified washing technique or absorbent paper strips. Following one week, the samples were recollected from the same tooth with another technique. The obtained samples were centrifuged to obtain cfDNA in the supernatant, from which DNA extraction was performed by using InstaGene Matrix. The concentration of cfDNA was measured by nanodrop spectrophotometer and processed to polymerase chain reaction with specific primers for 110, 536, and 2,000 bp of human β-globin gene.
Results: There was no significant difference in cfDNA concentration between the two collection techniques.
Furthermore, these cfDNA concentrations were not significantly correlated with any clinical periodontal
parameters. Interestingly, 2,000 bp PCR products showed positive correlation among all clinical periodontal
parameters (p<0.05). The presence of this PCR product was significant to the level of severity of the clinical
parameters.
Conclusion: The cell free DNA collected by absorbent paper strips was as good as by modified washing technique. Therefore, the absorbent paper strips could be used for GCF collection instead of the modified washing technique. The presence of 2,000 bp PCR product, in GCF may be related to the severity of periodontal diseases.
Article Details
References
Genco RJ, Slots J. Host responses in periodontal diseases. J Dent Res 1984; 63: 441-51.
Board of Trustees on American Academy of Periodontology. American Academy of Periodontology Task Force Report on the Update to the 1999 Classification of Periodontal Diseases and Conditions. J Periodontol 2015; 86: 835-8.
Board of Trustees on American Academy of Periodontology. Parameter on comprehensive periodontal examination. J Periodontol 2000; 71: 847-8.
Jahr S, Hentze H, Englisch S, Hardt D, Fackelmayer FO, Hesch RD, et al. DNA fragments in the blood plasma of cancer patients: quantitations and evidence for their origin from apoptotic and necrotic cells. Cancer Res 2001; 61: 1659-65.
Lo YM, Corbetta N, Chamberlain PF, Rai V, Sargent IL, Redman CW, et al. Presence of fetal DNA in maternal plasma and serum. Lancet 1997; 350: 485-7.
Elshimali YI, Khaddour H, Sarkissyan M, Wu Y, Vadgama JV. The clinical utilization of circulating cell free DNA (CCFDNA) in blood of cancer patients. Int J Mol Sci 2013; 14: 18925-58.
Kustanovich A, Schwartz R, Peretz T, Grinshpun A. Life and death of circulating cell-free DNA. Cancer Biol Ther 2019; 20: 1057-67.
Bronkhorst AJ, Ungerer V, Holdenrieder S. The emerging role of cell-free DNA as a molecular marker for cancer management. Available from https://dx. doi.org/ 10.1016/j.bdq.2019.100087. online 18 March 2019.
Remijsen Q, Kuijpers TW, Wirawan E, Lippens S, Vandenabeele P, Vanden Berghe T. Dying for a cause: NETosis, mechanisms behind an antimicrobial cell death modality. Cell Death Differ 2011; 18: 581-8.
Sima C, Glogauer M. Neutrophil Dysfunction and Host Susceptibility to Periodontal Inflammation: Current State of Knowledge. Curr Oral Health Rep 2014; 1: 95-103.
Vitkov L, Klappacher M, Hannig M, Krautgartner WD. Extracellular neutrophil traps in periodontitis. J Periodont Res 2009; 44: 664-72.
Bronkhorst AJ, Wentzel JF, Aucamp J, van Dyk E, du Plessis L, Pretorius PJ. Characterization of the cell-free DNA released by cultured cancer cells. Mol. Cell 2016; 1863: 157-65.
Al-Humood S, Zueriq R, Al-Faris L, Marouf R, Al-Mulla F. Circulating cell-free DNA in sickle cell disease: is it a potentially useful biomarker?. Arch Pathol Lab Med 2014; 138: 678-83.
Antonio F, Akihiko S, Mariko I, Ryu M, Kiyotake I, Takashi O. Total cell-free DNA (β-globin gene) distribution in maternal plasma at the second trimester: a new prospective for preeclampsia screening. Prenat Diagn 2004;24:722-6.
Thaweboon B, Laohapand P, Amornchat C, Matsuyama J, Sato T, Nunez PP, et al. Host betaglobin gene fragments in crevicular fluid as a biomarker in periodontal health and disease. J Periodontal Res 2010; 45: 38-44.
Doan TM, Kriangcherdsak Y, Thaweboon S, Thaweboon B. Evaluation of host beta-globin gene fragment lengths in peri-implant crevicular fluid during the wound healing process: a pilot study. Int J Oral Max Impl 2015; 30: 1295-302.
Nazar Majeed Z, Philip K, Alabsi AM, Pushparajan S, Swaminathan D. Identification of gingival crevicular fluid sampling, analytical Methods, and oral biomarkers for the diagnosis and monitoring of periodontal diseases: a systematic review. Available from https://doi.org/10.1155/2016/1804727. online 15 December 2016.
Armitage GC. Development of a classification system for periodontal diseases and conditions. Ann Periodontol 1999; 4: 1-6.
Trigg RM, Martinson LJ, Parpart-Li S, Shaw JA. Factors that influence quality and yield of circulatingfree DNA: a systematic review of the methodology literature. Available from https://doi.org/10.1016/j. heliyon.2018.e00699. online 25 July 2018.
Sherwood JL, Corcoran C, Brown H, Sharpe AD, Musilova M, Kohlmann A. Optimised pre-analytical methods improve KRAS mutation detection in circulating tumour DNA (ctDNA) from Patients with non-small cell lung cancer (NSCLC). Available from https://doi.org/10.1371/journal.pone.0150197. online 26 February 2016.
Magán-Fernández A, O'Valle F, Abadía-Molina F, Muñoz R, Puga-Guil P, Mesa F. Characterization and
comparison of neutrophil extracellular traps in gingival samples of periodontitis and gingivitis: A pilot study. J Periodontal Res 2019; 54: 218-24.
Brusca SB, Elinoff JM, Jang MK, Demirkale CY, Valantine HA, Solomon MA, et al. Plasma cell-free DNA as a novel marker of disease severity in pulmonary arterial hypertention. Available from https://doi.org/10.1016/S0735-1097(19)32503-3. online 17 March 2019.
Yu D, Tong Y, Guo X, Feng L, Jiang Z, Ying S, et al. Diagnostic value of concentration of circulating cell-free DNA in breast cancer: a meta-analysis. Available from https://dx.doi.org/10.3389/fonc. 2019.00095. online 1 March 2019.
Karlas T, Weise L, Kuhn S, Krenzien F, Mehdorn M, Petroff D, et al. Correlation of cell-free DNA plasma concentration with severity of non-alcoholic fatty liver disease. Available from https://doi.org/10.1186/ s12967-017-1208-6. online 19 May 2017.
Hamaguchi S, Akeda Y, Yamamoto N, Seki M, Yamamoto K, Oishi K, et al. Origin of circulating free DNA in sepsis: analysis of the CLP Mouse Model. Available from https://doi.org/10.1155/2015/614518. online 26 July 2015.
Beck JD, Cusmano L, Green-Helms W, Koch GG, Offenbacher S. A 5-year study of attachment loss in community-dwelling older adults: incidence density. J Periodontal Res 1997; 32 : 506-15.
Mdala I, Olsen I, Haffajee AD, Socransky SS, Thoresen M, de Blasio BF. Comparing clinical attachment level and pocket depth for predicting periodontal disease progression in healthy sites of patients with chronic
periodontitis using multi-state Markov models. J Clin Periodontol 2014; 41: 837-45.