CATEGORY 84 AUSTRALASIAN DENTIST CLINICAL the site of tension and resorbed at the site of compression. A longitudinal study was performed to assess the level of alkaline phosphatase within GCF in a patient undergoing orthodontic treatment26. It was noted that the enzyme level was elevated at the site of tension, as compared to the site of compression. Hence, these enzymes represent future novel markers and are associated with gingival in ammation caused during forces applied by orthodontic appliances. Orthodontics demands accurate timings for the management of most skeletal discrepancies; di erent stages of bone growth and its maturation in the subject must be known. is is usually achieved most commonly by identifying and comparing chronological age and dental age, and by CVM staging and handwrist radiograph. e former methods are relatively unreliable, and the latter methods involve radiation exposure27. Hence, any method that is noninvasive with minimum side e ects and that can help the clinician to identify the growth spurt is in highly demand. Research that showed variable peptides in the GCF of both prepubertal and post-pubertal subjects has been conducted recently28. Further studies are encouraged to nd more information about these novel biomarkers. 3. GCF Proteomic Analysis After collection, GCF samples will go through di erent steps of proteomics analysis, as illustrated in Figure 4. A variety of proteolytic enzymes has been identi ed in GCF, such as collagenase, elastase, and cathepsin B, D, H, and L29. ese proteolytic enzymes are reported as the destructor of periodontal tissues and have the capability of degrading type-I collagen and glycoproteins30. e most commonly reported identi ed proteins from GCF are actin, keratins, histones, annexins, proteins S100-A9, apolipoprotein A-1, albumin, salivary gland antimicrobial peptides (histatins, HNP-1, -2 & -3, LL-37, statherin), and cystatin B [31,32]. Some immunerelated proteins present in GCF include Ig gamma-1 chain C region, Ig gamma-3 chain C region, lactoferroxin-C, leukocyte elastase inhibitor, alpha 1 antitrypsin, heat shock protein beta-1, and coronin-1A33. Table 2 presents a detailed pro ling of GCF proteins, proteomic tools used, and some of the proteins identi ed. 4. Conclusions Gingival crevicular uid is a serum exudate that originates from the periodontal sulcus or pocket and is regarded as a promising biological uid for the detection of periodontal disease. Its composition resembles normal serum, but its volume uctuates in certain conditions such as those of gingivitis, caries, external root resorption, and chronic periodontitis, as well as during orthodontic tooth movement. GCF is composed of variable substances that include immunoglobulin, enzymes, local mediators, toxin cells, protein peptides, tissue breakdown products, and microorganisms. e level of this substance when uctuating in the above-mentioned conditions, as reported in many papers, will mark as a future diagnostic tool in their non-invasive analysis. Due to limitations in its collection, which includes volume size and contamination, collecting methods need further work, and a way to improve the ease for clinicians must be found; such development would help us to better demonstrate the pathogenesis of such diseases and to determine better strategies for treatment and early prevention. u Acknowledgments: We are thankful to Pakistan Human Saliva Research Group (PakSRG) for guiding us in the compilation of this whole manuscript. Author Contributions: All authors work equally. Con icts of Interest: e authors declare no con ict of interest. Contact gapmagazines@gmail.com for a complete list of references. Dent. J. 2017, 5, 12 6of 8 Table 2. Description of reported studies on GCF biomarkers analysis. Author& Year Sample Collection Sites Collection Method Proteomic Tool Number of Identified Proteins Outcome of Study Ref. Baliban et al., 2012 Collected from pre-selected sites with probing depth >6 mm and <8 mm in periodontitis patients and for periodontal health from mesio-buccal sites of first molar Filter strips (Periopapers®, Interstate Drug Exchange, Amityville, NY, USA) Protein digest with trypsin, HPLC, fragmented analysis with tandem mass spectrometry (MS/MS) 432 human proteins identified (120 new) Studied identified novel biomarkers from GCF of periodontal healthy and chronic periodontitis patients [31] S.Tsuchida et al., 2012 Labial side of maxillary incisors without crown and restoration Absorbent paper points (ZIPPERER®,Munich, Germany) 2DE, SDS-PAGE, Western Blot analysis, HPLC with LTQ-XL, HPLC with LTQ-Orbitrap XL, LC-MS/MS 327 proteins identified SOD1 and DCD were significantly↑ in GCF of periodontal patients [14] Carneiro et al., 2012 Healthy gingival sulcus of the second and third molar teeth Periopapers®,USA Trypsin digested followed by nano-flow liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) analysis and enzyme-linked immunosorbent essay (ELISA) for human albumin analysis 199 proteins identified Provide protein analysis of healthy periodontium and explore GCF composition with new groups of proteins specific to GCF microenvironment [18] Ngo et al., 2013 Five deepest sites and molar sites except mesial surface were excluded Microcaps (glass micocapillary tubes); Drummed Scientific, Brookmall, PA, USA Matrix-assisted laser desposition/ionization time-of-flight (MALDI-TOF) mass spectrometry - GCF mass spectra could be best for analyzing attachment loss and diagnosis of periodontal diseases [32] CarinaM., et al. 2013 Chronic Periodontitis patients sample were taken from different sites (5 deep sites, 5 shallow sites with gingivitis, and 4 without bleeding on probing sites) Periopaper strip (ProFlow Inc., Amityville, NY,USA) Reversed-phase LC-ESi-MS/MS andELISA 230 proteins identified Concluded marked differences in GCF proteomics in different disease profiles [33] Carneiro et al., 2014 The pre-selected specific sites with moderate and severe chronic periodontal disease were defined by pocket depth of 5–7 mm (24 patients) and >7 mm (16 patients) Periopaper strips (Oraflow, Plainview, NY,USA) SDS-PAGE, Isotope-Coded-Affinity-Tag (ICAT) labeling, mTRAQ labeling, Nano-LC-ESI-MS/MS, Human Albumin ELISA Kit, and S100-A9 protein quantification by ELISA 199 proteins Identified Innovative approach concluded the novel changes in host and microbial derived GCF proteome of periodontal patients [19] Rody, Jr. et al., 2014 Collected from a deciduous second molar with radiographic evidence of root resorption on 1 quadrant (experimental site) and from the permanent first molar on the contralateral quadrant (control site) in the same jaw. Periopaper strips (Oraflow, Plainview, NY,USA) One dimensional LC-MS and Two dimensional LC-MS 2789 proteins in control group and 2421 proteins in root resorption group Mass spectrometry is a useful tool for analyzing external root resorption [23] Kinney et al., 2014 Collection from the mesio-buccal aspect of each site (tooth) for up-to 28 teeth per patient. Methylcellulose strip (Pro Flow, Inc., Amityville, NY,USA) ELISA and Quantibody Human Cytokine Array - This method offers improved patient monitoring and disease control [21] Huynh et al., 2015 Collection were chosen based on how well they represented the healthy, gingivitis, and chronic periodontitis inclusion criteria Glass-microcapillary tube (Drummond Scientific, Brookmall, PA, USA) One-dimensional Gel Electrophoresis and Nano-LC-ESI-MS 121 proteins identified Concluded various biomarkers which differentiate between healthy periodontium, gingivitis, and chronic periodontitis [22] Figure 4. A flow diagram representing the proteomic analysis of healthy and diseased conditions via gingival crevicular fluid (GCF), adapted from Khurshid et al. [16]. Table 2. Description of reported studies on GCF biomarkers analysis.
RkJQdWJsaXNoZXIy MTc3NDk3Mw==