Table 2.

Evidence Related to Study of Various Biomarkers and Proteins in Various Biofluids Associated to EARR.

Authors Experimental Subjects/Teeth (No./Age/Sex) Biomarkers Studied Condition Analyzed Detection of Root Resorption Controls/Teeth (No./Age/Sex) Medium Studied Technique Outcomes Conclusions
2004 Mah et al.13 Grp 1: Mx central incisor (n=20) with 1–3 mm RR; 13F, 7M; 12–16 y
Grp 2: 10 second molars (n=20); 15F, 5M; 9–12 y
DPP Orthodontic Tx (not specified) Radiographs (not specified) Mx central incisors (n=20) of untreated pts, 12F, 8M, 12–16 y GCF Periopaper, ELISA Levels of DPP: greatest in resorbing 10 molar (11.7±4.1 μg/mg) followed by orthodontically treated tooth (9.3±4.7 μg/mg) and least in controls (5.4±4.1 μg/mg); NS between resorption Grps DPP can be detected in exfoliating primary teeth and orthodontic root resorption
2007 Balducci et al.7 20 pts with mild RR (≤2 mm) (11F, 9M, 14–40 y), 20 pts with severe RR (>2 mm) (15F, 5M, 15–44 y) DMP1, PP, DSP RR in orthodontic pts IOPA 20 pts (13F, 7M, 12–34 y); no RR/orthodontic Tx GCF Periopaper (mesial and distal of Mx central and lateral incisors), SDS–PAGE, stained western blot, ELISA Molecular weight 77, 66, 55, 50, and 26 kDa proteins identified, NS between control and study Grps in immunoblot; ELISA showed Sig. ↑ of DMP1, PP, DSP in RR vs control Grps and of PP and DSP in severe RR vs mild RR Grps DMP1, DSP, and PP in GCF proved a biomarker for RR in orthodontic Tx
2008 Kereshanan et al.5 Grp 1: 50 second 10 molars (9–14 y) (advanced coronal RR [n=33] and apical minimal RR Grp [n=17])
Grp 2: 20 pts (11–15 y), T0=pre-fixed Tx, T1=12 mo post start of Tx
DSP Physiological RR and OTM Orthopantomogram Control: 20 pts (10–15 y) erupted second premolars with no RR GCF Micropipettes, slot blot immunoassay, DSP in dentin of 10 molars by western blot DSP levels: greater in physiological RR than non-resorbing teeth, DSP levels NS between coronal RR and apical RR; DSP levels Sig. higher in T1 compared to T0 DSP in GCF proved a biomarker of root resorption
2009 George et al.19 Grp 1: mild RR of 2 mm (20 pts, Tx 1 y)
Grp 2: severe RR >2 mm (20 pts)
OPN, OPG, RANKL Orthodontic Tx (not specified) Radiographs (not specified) 20 pts: no Tx, no RR GCF Periopaper (mesial/distal of Mx central and lateral incisors), SDS-PAGE, western blot Proteins conc greater in severe RR (0.89 μg/μL ±0.32 μg) than mild RR (0.77 μg/μL ±0.21 μg) and least in controls (0.22 μg/μL ±0.05 μg); ELISA showed Sig. higher RANKL antibodies in RR Grps than control Grp; RANKL/OPG ratio in severe RR Sig. greater than in control Grp Presence of OPN, OPG, and RANKL in root resorption
2013 Kunii et al.9 5 pts with severe RR (5F, mean age 28.9±6.1 y; mean orthodontic Tx duration of 27.8±3.3 mo) IL-6 All 4 extr orthodontic Tx Radiographs (not specified) 15 pts without RR (13F, 2M, mean age, 28.0±5.3 y; mean orthodontic Tx duration of 26.4±3.1 mo) GCF Periopaper (mesial/distal of Mx central and lateral incisors), ELISA IL-6 protein levels Sig. ↑ in RR than non-RR Grp IL-6 in GCF proved a root resorption biomarker in orthodontic Tx
2013 Wahab et al.15 12F (Mx canines as test teeth), 100 g/150 g force to either side, split mouth design ALP Class II div 1 malocc; upper 4/4 extr with retraction by NiTi coil spring IOPA Mand canine as control GCF, collection weekly for 6 wk Periopaper (mesial/distal of Mx canine, Mand canine), spectrophotometry at 405 nm ALP at mesial sites peak at wk 1 showing Sig. diff with 100 g force; no RR for test/control teeth in 150/100 g force Canine movement greater with 150 g than 100 g force and higher ALP at mesial sites with no RR
2014 Sha et al.2 20 pts (12F, 8M, 13–24 y), 8–12 mo of orthodontic Tx DSPP Orthodontic Tx (not specified) Radiographs (not specified) Same pts for both methods (ELISA with spectrophotometry and electrochemical detection) GCF Filter paper strip (mesial/distal sites of left and right Mx central incisors), ELISA DSPP detection with spectrophotometric ELISA 10 times greater than with electrochemical detection. DSPP conc range NS between methods DSPP can be sensitively and accurately detected in root resorption
2014 Vieira20 Total 60 pts (38F, 22M, 15–30 y with orthodontic Tx of 6 mo); Grp 2: 30 pts, mild to moderate RR Proteins Orthodontic Tx (not specified) IOPA Grp 1: 30 pts, no RR GCF Sterile absorbent paper cones, 2-DE gels, SDS-PAGE with isoelectric focusing Greatest sharpness to detect protein bands with Milli-Q ultrapure ice-cold water, without GCF protein extraction Protein extraction protocols tested for accuracy
2014 Rody et al.21 11 pts (7F, 4M, 10–11 y) second 10 molars with RR in one quadrant; split mouth design Proteins No orthodontic Tx Radiograph (not specified) 11 pts (7F, 4M, 10–11 y), permanent 1st molar on contralateral side with no RR GCF Periopaper (lingual side of 10 and permanent molars), LC-MS, nano-flow LC system coupled to triple TOF 5600 MS Total 37 RR proteins upregulated and 59 RR proteins downregulated RR proteins upregulation and downregulation identified in RR
2015 Wahab et al.22 19 (13F, 6M), split mouth design, either 100 g or 150 g force ALP, TRAP, AST All 4 extr and retr IOPA Internal control (baseline) GCF, Baseline (0 wk), 1–5 wk Periopaper (mesial/distal of Mx right and left canine), spectrophotometry 100 g Grp: TRAP Sig. ↑ from baseline to 3–5 wk and slight rise of ALP, AST from baseline; 150 g Grp: ALP, TRAP activities ↑ slightly from baseline, AST Sig. ↑ in 5 wk 150 g force and 100 g force show NS difference in AST, ALP, or TRAP levels
2016 Lombardo et al.23 6 pts (5F, 1M), average age 14 y, 12 wks orthodontic Tx DSP Orthodontic treatment with Damon appliances Radiographs (not specified) Same pts for both methods (conventional ELISA vs DSP antibody-coated magnetic micro-beads prior to ELISA) GCF Mesial and Ds sites of Mx central and lateral incisors, sterile paper strips Sig. diff between standard ELISA and micro-beads for DSP evaluation in early RR evaluation; results of micro-bead approach are more uniform and highly sensitive Modified micro-bead approach is more reliable for early detection of RR for DSP evaluation
2016 Rody et al.16 11 pts (7F, 4M, 10–11 y), second 10 molars with RR in one quadrant IL-1β, IL-1RA, MMP-8, DSP, RANKL, OPG No orthodontic Tx Radiograph (not specified) Permanent 1st molar on contralateral side with no RR GCF Lingual side of 10 and permanent molars, Periopaper, immunoassay NS in IL-1β, OPG, or MMP-9 between exp and control Grp; RANKL data unreliable; IL-1RA Sig. downregulated in RR IL-1RA down-regulation in GCF from 10 molars with root resorption
2017 Yashin et al.12 9 pts (mean age 23±2.9 y), moderate to severe RR Cytokine profile in saliva Finished orthodontic Tx within 2 y Orthopantomograms Pts with no RR Blood and saliva 10 mL unstimulated saliva collected by expectoration, ELISA Saliva: moderate to severe RR show Sig. ↑ in IL-7, IL-10, IL-12p70, and IFN-γ, Sig. ↓ in IL-4; blood: control group has higher osteocalcin and P1NP than RR Saliva can be used for cytokine assessment in root resorption
2017 Kaczor-Urbanowicz et al.24 48 pts with RR (31F, 17M)
Grp 1: moderate to severe RR young pts (11F, 6M); Grp 2: moderate to severe RR adult pts (7F, 4M); Grp 3: mild RR young pts (7F, 4M); Grp 4: mild RR adult pts (6F, 3M)
Proteins Not specified IOPA (Mx central and lateral incisors) at T0 (before bonding), T9 (9 mo after bonding) 24 pts without RR (13F, 11M)
Grp 5: control young pts (7F, 6M); Grp 6: control adult pts (6F, 5M)
Saliva Unstimulated whole saliva, 2D gel electrophoresis, quantitative mass spectrometry, western blot 772 proteins identified by qMS, 244 highly increased expression profile, Sig. ↑ in moderate to severe young RR Grp compared to controls and 58 proteins in the adult Grp Salivary proteins associated with root resorption identified
2017 Thalanany et al.25 20 pts, 13–22 y; exp Grp: 10 pts undergoing orthodontic Tx DSPP Simultaneous intrusion and retr arch Radiograph (not specified) Control Grp: no orthodontic Tx GCF, T0= before intrusion, T1=2 mo after intrusion Mx right and left central and lateral incisors; microcapillary tubes, ELISA Sig. ↑ in DSPP at T1 compared to T0 DSPP may be marker for root resorption
2017 Ahuja et al.26 8 (2F, 6M, age range 13.9–22.9 y)
Split mouth design: test vs control sides
IL-1β, 2, 4, 5, 6, 7, 8, 10, 12, 13, INF-γ, TNF-α, GM-CSF 225 g buccal tipping force for 28 d on test side Micro-CT Contralateral teeth (control side) GCF, Time points: 0 h (prior to force), 3 h, 1 d, 3 d, 7 d, 28 d Periopaper, multiplex bead immunoassay IL-1β: Sig. ↑ peak at days 1 and 7 but NS between test and control side; IL-4: ↑, peak days 1–3; IFN-γ: peak at 72 h; TNF-α: ↑ at 3 h, 28 d; IL-7 peak at 28 d; GM-CSF: immediate ↓, ↑ at 7 d, peak at 28 d; Comparison between low and high RR: GM-CSF show Sig. ↑ in low RR; Micro-CT: mesial, distal surface, and middle 3rd showed sig. ↑ RR on test side teeth Pro-resorptive cytokines (IL-7, TNF-α) ↑ in high orthodontic forces, anti- resorptive cytokines (GM-CSF) ↓ initially
2018 Zhou et al.27 8F pts with RR, mean age 22.25 y, Tx duration 22.37 mo Metabolites Both extr and non-extr Tx Orthopantomograms 11F controls, mean age 24.27 y, Tx duration 21 mo Saliva Unstimulated saliva collected from occlusal space of right Mand molars without chewing for ~3 min 187 metabolites identified, including butyrate, propane-1,2-diol, α-linolenic acid (Ala), α-glucose, urea, fumarate, formate, guanosine, and purine Difference in metabolites in saliva of RR pts can be detected by 1 HNMR-based metabolomics method
2020 Mohd Nasri et al.28 10 pts Protein abundance Mx and Mand fixed appliances IOPA at T0 and T6 of Mx central incisors None GCF at T0 (pre-Tx), T1 (1 mo), T3 (3 mo), T6 (6 mo) Mesial and Ds of Mx central incisors Periopaper, liquid chromatography-tandem mass spectrometry Increased protein abundance of S100A9, immunoglobulin J chain; heat shock protein 1A, immunoglobulin heavy variable 4–34 and vitronectin at T1; protein abundance ↑ of thymidine phosphorylase at T3 Early RR protein markers identified
2021 Mandour et al.29 74 pts (3 Grps: 2 Tx, 1 control)

Grp 1: orthodontic pts (1–3 mm RR); Grp 2: pediatric pts (lower second 10 molars, physiologic RR)
IL-1RA, DSPP Not specified Radiograph (not specified) Grp 3: control (no RR, no orthodontic Tx) GCF Endodontic paper points, ELISA IL-1RA levels in controls greater than orthodontic pts, and least in pediatric Grp; DSPP levels in pediatric group higher than in orthodontic pts, and least in controls; IL-1RA cut-off for OIRR (≤432.6 pg/mL) and DSPP (≥7.33 pg/mL); DSPP reliability (100%) vs IL-1RA (80%) IL-1RA, DSPP biomarkers for OIRR
2020 Zain et al.30 7 orthodontic pts, 2 samples taken at 3 and 6 mo into orthodontic Tx and 3 samples at 12 mo of orthodontic Tx DSPP Fixed orthodontic Tx Not specified 3 non-orthodontic control samples GCF Mx central incisors, Periopaper, absorption spectroscopy Control sample showed lower peak in absorption spectrum than exp sample (3, 6, 12 mo); spectrum proportional to Tx duration, 0.91 accuracy Higher absorption spectrum of DSPP indicates higher resorption

↑, greater/increase; ↓, lower/decrease; 1 HNMR, hydrogen-1 nuclear magnetic resonance; 10, primary; ALP, alkaline phosphatase; AST, aspartate aminotransferase; conc, concentration; CT, computed tomography; d, day(s); diff, difference; DMP1, dentin matrix protein 1; DPP, dentin phosphophoryn; Ds, distal; DSP, dentin sialoprotein; DSPP, dentin sialophosphoprotein; EARR, external apical root resorption; ELISA, enzyme-linked Immunosorbent assay; exp, experimental; extr, extractions; F, female(s); GCF, gingival crevicular fluid; GM-CSF, granulocyte-macrophage colony-stimulating factor; Grp, group(s); h, hour(s); IFN-γ, interferon gamma; IL, interleukin; IL-RA, interleukin-1 receptor antagonist; IOPA, intraoral periapical radiograph; LC, liquid chromatography; LC-MS, liquid chromatography-mass spectrometry; M, male(s); malocc, malocclusion; Mand, mandibular; min, minute(s); MMP, matrix metalloproteinase; mo, month(s); MS, mass spectrometry; Mx, maxillary; NS, no statistically significant difference; OIRR, orthodontically induced root resorption; OPG, osteoprotegerin; OPN, osteopontin; OTM, orthodontic tooth movement; P1NP, procollagen type 1 N-terminal propeptide; PP, dentin phosphophoryn (alternate abbreviation in the literature); pts, patients; qMS, quadrupole mass analyzer; RANKL, receptor activator of nuclear kappa B ligand; retr, retraction; RR, root resorption; SDS-PAGE, Sodium dodecyl-sulfate polyacrylamide gel electrophoresis; Sig., significant; TNF-α, tumor necrosis factor-α; TOF, time of flight; TRAP, tartrate-resistant acid phosphatase; Tx, treatment; vs, versus; wk, week(s); y, year(s).

RMMJ Rambam Maimonides Medical Journal Rambam Health Care Campus 2022; 13(4): e0027. ISSN: 2076-9172
Published online 2022 October 27. doi: 10.5041/RMMJ.10482