Proteomic Analysis of Gingival Crevicular Fluid During Tooth Eruption

Year 2023, , 863 - 870, 29.12.2023

Selin Yıldırım Albat , Serap Akyüz , Ayşen Yarat , Fikrettin Şahin , Hüseyin Çimen

https://doi.org/10.33808/clinexphealthsci.1214974

Abstract

Objective: Gingival crevicular fluid (GCF) is a biological fluid that has the unique capacity to reflect changes in periodontium to its protein composition, making it ideal for potential biomarkers. There is limited information about the mechanism of tooth eruption, for which GCF might provide valuable knowledge. This study aimed to provide a proteomic approach to investigate the composition of GCF obtained from two different supraosseous tooth eruption stages of permanent molars changes.
Methods: GCF samples were taken from a total of 26 healthy children, whose permanent molar just emerged from the gingiva (5-8 years old) and the occlusal equilibrium stage (9-13 years old). Proteins were extracted with Bio-Rad Rehydration Buffer followed by ZebaTM Spin Desalting Column. GCF samples were separated with two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) followed by mass spectrometry- based protein identification.
Results: A new optimized protocol with enriched protein extraction from GCF samples was developed. Several proteins related to tooth eruption were detected. It was determined that keratin type II cytoskeletal 4 and keratin type I cytoskeletal 9 decreased and albumin increased in GCF protein content in erupting teeth compared to those in the occlusal equilibrium phase.
Conclusion: This methodology, which we have applied for 2D-PAGE of GCF, can also be a source for other studies. There is huge diagnostic potential in mass spectrometry technologies, this study can be carried forward by using other approaches.

Keywords

Tooth eruption, Gingival crevicular fluid, Proteomic analysis, Two-dimensional gel electrophoresis, Mass spectrometry

Supporting Institution

Marmara University Scientific Research Projects Commission

Project Number

SAG-C-DRP-080519-0181

Thanks

This study was funded by Marmara University Scientific Research Projects Commission (Project-No: SAG-C-DRP-080519-0181).

References

• Massler M. Studies in tooth development: Theories of eruption. Am J Orthod. 1941;(27):552-576.
• Steedle JR, Proffit WR. The pattern and control of eruptive tooth movements. Am J Orthod. 1985;87(1):56-66. DOI: 10.1016/0002-9416(85)90174-5.
• Craddock HL, Youngson CC. Eruptive tooth movement-the current state of knowledge. Br Dent J. 2004;197(7):385-391. DOI: 10.1038/sj.bdj.4811712.
• Carneiro LG, Venuleo C, Oppenheim FG, Salih E. Proteome data set of human gingival crevicular fluid from healthy periodontium sites by multidimensional protein separation and mass spectrometry. J Periodontal Res. 2012;47(2):248-262. DOI: 10.1111/j.1600-0765.2011.01429.x.
• Carneiro LG, Nouh H, Salih E. Quantitative gingival crevicular fluid proteome in health and periodontal disease using stable isotope chemistries and mass spectrometry. J Clin Periodontol. 2014;41(8):733-747. DOI: 10.1111/jcpe.12262.
• Vernerová A, Krčmová LK, Heneberk O, Radochová V, Švec F. Liquid chromatography method with tandem mass spectrometry and fluorescence detection for determination of inflammatory biomarkers in gingival crevicular fluid as a tool for diagnosis of periodontal disease. J Pharm Biomed Anal. 2022;212:114644. DOI: 10.1016/j.jpba.2022.114644.
• Bostanci N, Heywood W, Mills K, Parkar M, Nibali L, Donos N. Application of label-free absolute quantitative proteomics in human gingival crevicular fluid by LC/MS E (gingival exudatome). J Proteome Res. 2010;9(5):2191-2199. DOI: 10.1021/pr900941z.
• Silva-Boghossian CM, Colombo AP, Tanaka M, Rayo C, Xiao Y, Siqueira WL. Quantitative proteomic analysis of gingival crevicular fluid in different periodontal conditions. PLoS One. 2013;8(10):e75898. DOI: 10.1371/journal.pone.0075898.
• Iavarone F, Olianas A, Patini R, Gallenzi P, Di Tonno L, Desiderio C, Cabras T, Manconi B, Vincenzoni F, Cordaro M, Messana I, Urbani A, Castagnola M. Top down proteomic analysis of gingival crevicular fluid in deciduous, exfoliating and permanent teeth in children. J Proteomics 2020;226:103890. DOI: 10.1016/j.jprot.2020.103890.
• Ozkavaf A, Aras H, Huri CB, Yamalik N, Kilinc A, Kilinc K, Caglayan F. Analysis of factors that may affect the enzymatic profile of gingival crevicular fluid: Sampling technique, sequential sampling and mode of data presentation. Journal of Oral Science 2001;43(1):41-48. DOI: 10.2334/josnusd.43.41.
• Bio-Rad. PDQuest™. User Guide for Version 7.4.0. Published [2005] Accessed [30.11.2022] https://www.bio-rad.com/webroot/web/pdf/lsr/literature/10002941.pdf
• Kjær I. Mechanism of human tooth eruption: Review article including a new theory for future studies on the eruption process. Scientifica. 2014;2014:341905. DOI: 10.1155/2014/341905.
• Cahill DR, Marks SC Jr. Tooth eruption: Evidence for the central role of the dental follicle. J Oral Pathol. 1980; 9(4):189-200. DOI: 10.1111/j.1600-0714.1980.tb00377.x.
• Marks Jr SC, Cahill DR. Experimental study in the dog of the non-active role of the tooth in the eruptive process. Arch Oral Bio. 1984; l29(4): 311-322. DOI:10.1016/0003-9969(84)90105-5
• Marks SC, Cahill DR. Regional control by the dental follicle of alterations in alveolar bone metabolism during tooth eruption. Journal of Oral Pathology. 1987;16(4):164–169. DOI: 10.1111/j.1600-0714.1987.tb02060.x.
• Lin F, Fan W, Wise GE. Granule proteins of the dental follicle and stellate reticulum inhibit tooth eruption and eyelid opening in postnatal rats. Arch Oral Biol. 1992;37(10):841-847. DOI: 10.1016/0003-9969(92)90118-r.
• Wise GE, Frazier-Bowers S, D'Souza RN. Cellular, molecular, and genetic determinants of tooth eruption. Crit Rev Oral Biol Med. 2002;13(4):323-334. DOI: 10.1177/154411130201300403.
• Wise GE, Yao S. Regional differences of expression of bone morphogenetic protein-2 and RANKL in the rat dental follicle. Eur J Oral Sci. 2006;114(6):512-516. DOI: 10.1111/j.1600-0722.2006.00406.x.
• Wise GE, King GJ. Mechanisms of tooth eruption and orthodontic tooth movement. J Dent Res. 2008;87(5):414-434. DOI: 10.1177/154405910808700509.
• Nishijima Y, Yamaguchi M, Kojima T, Aihara N, Nakajima R, Kasai K. Levels of RANKL and OPG in gingival crevicular fluid during orthodontic tooth movement and effect of compression force on releases from periodontal ligament cells in vitro. Orthod Craniofac Res. 2006;9(2):63-70. DOI: 10.1111/j.1601-6343.2006.00340.x.
• Shetty A, Jain M, Sneha K, Shetty V, Rao S, Shetty A. Evaluation of interleukin 6 levels in gingival crevicular fluid and periodontal ligament on application of orthodontic forces. World. 2022;13(1): 17. DOI:10.5005/jp-journals-10015-1891.
• Baliban RC, Sakellari D, Li Z, DiMaggio PA, Garcia BA, Floudas CA. Novel protein identification methods for biomarker discovery via a proteomic analysis of periodontally healthy and diseased gingival crevicular fluid samples. J Clin Periodontol. 2012;39(3): 203-212. DOI: 10.1111/j.1600-051X.2011.01805.x.
• Ngo LH, Veith PD, Chen YY, Chen D, Darby IB, Reynolds EC. Mass spectrometric analyses of peptides and proteins in human gingival crevicular fluid. J Proteome Res. 2010;9(4):1683-1693. DOI: 10.1021/pr900775s.
• Grant MM, Creese AJ, Barr G, Ling MR, Scott AE, Matthews JB, Griffiths HR, Cooper HJ, Chapple IL. Proteomic analysis of a noninvasive human model of acute inflammation and its resolution: the twenty-one day gingivitis model. J Proteome Res. 2010;9(9):4732-4744. DOI: 10.1021/pr100446f.
• Tsuchida S, Satoh M, Umemura H, Sogawa K, Kawashima Y, Kado S, Nomura F. Proteomic analysis of gingival crevicular fluid for discovery of novel periodontal disease markers. Proteomics 2012; 12(13):2190-2202. DOI: 10.1002/pmic.201100655.
• Yi J, Shen Y, Yang Y, Shen C, Liu B, Qiao L, Wang, Y. Direct MALDI-TOF profiling of gingival crevicular fluid sediments for periodontitis diagnosis. Talanta 2021; 225:121956. DOI: 10.1016/j.talanta.2020.121956.
• Rody Jr WJ, Holliday LS, McHugh KP, Wallet SM, Spicer V, Krokhin O. Mass spectrometry analysis of gingival crevicular fluid in the presence of external root resorption. Am J Orthod Dentofacial Orthop. 2014; 145(6):787-798. DOI: 10.1016/j.ajodo.2014.03.013.
• Moriya Y, Obama T, Aiuchi T, Sugiyama T, Endo Y, Koide Y, Noguchi E, Ishizuka M, Inoue M, Itabe H, Yamamoto M. Quantitative proteomic analysis of gingival crevicular fluids from deciduous and permanent teeth. J Clin Periodontol. 2017;44(4):353-362. DOI: 10.1111/jcpe.12696.
• Wen X, Franchi L, Chen F, Gu Y. Proteomic analysis of gingival crevicular fluid for novel biomarkers of pubertal growth peak. Eur J Orthod. 2018; 40(4):414-422. DOI: 10.1093/ejo/cjx082.
• McLaughlin WS, Kirkham J, Kowolik MJ, Robinson C. Human gingival crevicular fluid keratin at healthy, chronic gingivitis and chronic adult periodontitis sites. J Clin Periodontol. 1996; 23(4):331-335. DOI: 10.1111/j.1600-051x.1996.tb00554.x.
• Huynh AH, Veith PD, McGregor NR, Adams GG, Chen D, Reynolds EC, Ngo LH, Darby IB. Gingival crevicular fluid proteomes in health, gingivitis and chronic periodontitis. J Periodontal Res. 2015; 50(5):637-649. DOI: 10.1111/jre.12244.
• Odanaka H, Obama T, Sawada N, Sugano M, Itabe H, Yamamoto M. Comparison of protein profiles of the pellicle, gingival crevicular fluid, and saliva: Possible origin of pellicle proteins. Biol Res. 2020; 53(1):3. DOI: 10.1186/s40659-020-0271-2.
• Bickel M, Cimasoni G, Andersen E. Flow and albumin content of early (pre-inflammatory) gingival crevicular fluid from human subjects. Arch Oral Biol. 1985;30(8):599-602. DOI: 10.1016/0003-9969(85)90079-2.
• Griffiths R, Barbour S. Lipoproteins and lipoprotein metabolism in periodontal disease. Clin Lipidol. 2010; 5(3):397-411. DOI: 10.2217/clp.10.27.
• Takemaru M, Sawada N, Aiuchi T, Itabe H, Yamamoto M. Comparison of protein profiles of gingival crevicular fluids collected from incisors, canines, and molars. Showa Univ J Med Sci. 2019; 31(4), 365-372. DOI:10.15369/sujms.31.365.