Table 2.
Extended
| Assessment Method |
Main Results |
| Outcome: Histomorphological Changes | |
| Light microscope Evaluated changes: degree and type of inflammation, fibrous tissue formation, necrosis, disruption of odontoblastic layer, vascular dilation and resorption of cementum or dentin Groups: 13–18 y (n = 11)—adolescents, 25–35y (n = 11)—adults | 1. No significant differences of the histologic parameters between the test and control teeth during the 7 d and 1 m test period in adolescents and adults, P > .05 2. Significant differences between adolescents and adults after 1m in type and degree of inflammation in the test teeth, P = .032 3. Odontoblastic aspiration and cementum or dentin resorption were observed in adolescents-group after 7 d and in adult group after 30 days |
| Light microscope Times: 10 d (short-time) and 171 d (long time) Evaluated changes: intrapulpal axons | 1. No significant differences were found in intrapulpal axons (either qualitatively or quantitatively) 2. Alteration in axons myelinated were found in teeth with short time (1.5% to 3.5%) 3. No inflammation was evident |
| Light microscope and sequential video prints Times: 1 and 5 d of culture Evaluated changes: growth of microvessels | 1. Significantly greater growth in 5 d and 10 d in the pulps of orthodontically moved teeth, P < .05 2. Significantly greater growth in the orthodontic group in all areas at 5 d but only in the crown and root areas at 10 d, P < .05 3. Increase in microvessel length after 8 d and thickness was greater after 14 d 4. Vessel degeneration began after 14 to 21 days and after 28 d had occurred in most of pulp |
| Inverted microscope and sequential video prints Times: 5, 10, and 14 d of culture Evaluated changes: growth of microvessels | 1. Significantly greater growth in 5, 10, and 14d in the pulps of orthodontically moved teeth, P < .05 |
| Inverted microscope and sequential video prints Times: 1 and 5 d of culture Evaluated changes: growth of microvessels | 1. No significant difference in the control teeth at 5 or 10 d of culture, P > .05 2. Significant reduction of pulpal microvessels (5 and 10 d) of test teeth group, P < .05 |
| Light microscope, immunohistochemistry and assessment with spectrophotometer Evaluated changes: odontoblastic and predentine layer, blood vessels and general pulp morphology; fibroblasts number; blood vessel analysis | 1. Vacuolation and degeneration of the odontoblast layer 2. The area of blood vessels increased after force 3. Blood vessel congestion and/or hemorrhage were not observed 4. Pulp stones were also observed occasionally 5. Inconsistencies in the pre-dentine layer were observed: with some sections having a regular predentine layer, while the pre-dentinal layer in other sections was highly irregular |
| Immunohistochemistry and assessment with spectrophotometer Evaluated changes: pulpal respiration rate | 1. Between all control and experimental premolars, there was a 27.4% depression in the respiration rate. 2. Between maxillary and mandibular premolars, there was a 31.52% and 23.28% depression in the respiration rate, respectively. |
| Light microscope Times: immediately before placement of the lingual button (T1–baseline), 1 wk after force (T2), 4 wk after force (T3), 8 wk after force (T4), 12 wk after force (T5) | 1. T2: observed moderate vascular congestion and the structure of the cell-free zone was disrupted in 300 g-force group 2. T3: presence of vacuolization in coronal pulp and resorption zones with cementum deposition in 300 g-force group 3. T4: vascular congestion and dilatation were more than T3. Presence of pulp stones 4. T5: presence of odontoblastic and adipose degeneration, vacuole formation in the coronal pulp of 300 g-force group |
| Light microscope Times: 1 and 3m of treatment time Evaluated changes: number of vessels, vessel areas, minimum vessel diameters, maximum vessel diameters, and predentine widths | 1. Vessel area and minimum and maximum vessel diameters showed significant differences among the groups after 1 mo, P < .05 2. Presence of pulp fibrosis zones after 3 mo |
| TEM | 1. Vascular degeneration was the main change in the pulps of experimental group 2. Presence of resorption areas in cementum of experimental group |
| Light microscope Evaluated changes: inflammatory response, soft and hard tissue response, and count of the number of blood vessels and pulp calcifications | 1. A significant increase of fibrous tissue, number of pulpal nodules and vasodilation in the experimental group were observed, P < .05 2. No significant differences in the number of blood vessels between the groups, P > .05 |
| Light microscope Times: before treatment (T0), 3 mo after (T1), 6 mo after (T2) | 1. In T0 and T1 the pulpal tissue did not show any significant morphological alteration 2. In T2, odontoblastic vacuolization was observed |
| Light microscope Times: 1 wk after force (T1), 2 wk after force (T2), 4 wk after force (T3) | 1. Certain characteristic pulpal reactions arise from orthodontic extrusion, these reactions involve: circulatory disturbances with congested and dilated blood vessels, odontoblastic degeneration, vacuolization and oedema of the pulp tissues, and (by the fourth week) manifestation of fibrotic changes |
| Light microscope Times: 3 d after force (T1), 3 wk after force (T2) Evaluated changes: degree and type of inflammation, fibrotic tissue formation, pulp stones and dentin formation, necrosis, disruption and vacuole formation in the odontoblastic layer, aspiration into the dentin tubules, cementum or dentin resorption, and vasodilation | 1. The extrusive and intrusive forces promoted disruption of odontoblastic layer and vacuolization after 3 d and 3 wk in comparison with control group, P < .05 2. The extrusive force promoted more fibrous tissue formation in comparison with control group and intrusive force after 3 wk, P = .01 3. Incomplete necrosis was observed in one tooth in extrusive group after 3 wk |
| Light microscope Times: 10 d after force (T1), 40 d after force (T2); Evaluated changes: inflammatory response, soft and hard tissue responses | 1. None of the teeth in the groups showed any inflammatory reactions or reparative dentin formation at the test periods. |
| Light microscope Times: 3 mo after RME (T1), 6 mo after RME (T2), 18 mo after RME (T3) Evaluated changes: number of vessels, vessel diameter, hemorrhage, vascular congestion, inflammatory cell infiltration, and vacuoles. | 1. Vessel diameter, hemorrhage, congestion and inflammatory cell infiltration varied between groups, and the differences between the control and 3 mo groups, and the 3 mo and 18 mo groups were most significant |
| Light microscope | 1. Deposition of secondary dentin in pulpal floor and down on the inner walls of the root canals after 4 mo 2. Presence of pulp stones in the root canals after 4 mo |
| Light microscope Evaluated changes: blood vessels, odontoblast layer, and presence of disruption or inflammation | 1. In the orthodontic group, there are increase in blood vessel area and vessels appeared to be dilated, congested when compared with the control group, P < .01 |
| Outcome: Tissue Factors Expression | |
| Radioimmunoassay Tissue factor: CGRP | 1. CGRP levels were higher in the severe force-group (0.1380 ± 0.0278 pmol/mg) than in the moderate force group (0.0609 ± 0.0236 pmol/mg), P < .0001 2. Significant statistical differences were found between control group and the severe force group, but not with the moderate force group and control group |
| Radioimmunoassay Tissue factors: Substance P, CGRP, b-Endorphin, and ME | 1. Significant differences in substance P levels between the control-group (83.51 ± 11.35 pmol/mg) and orthodontic-group (131.91 ± 26.32 pmol/mg), P < .05 |
| Immunohistochemical and assessment with spectrophotometer, real-time PCR Times: control (T0), 1 wk after (T1), 4 wk after (T2), 8 wk after (T3), 12 wk after force (T4) Tissue factors: c-Fos and MMP-9 | 1. C-Fos and MMP-9 PCR analysis showed that protein expression increased after 4 wk of treatment, with statistical difference significant between T0 and T2, T3, T4, P < .05 2. There was appositive correlation between c-Fos and MMP-9 expression, P < .001 |
| Immunohistochemical and assessment with spectrophotometer Times: after 7 d (T1), after 14 d (T2), after 3 mo (T3), after 6 mo (T4), after 14 mo (T5) Tissue factors: nNOS and iNOS | 1. The results suggest a close correlation between the duration of the orthodontic force and the expression of NOS. 2. The presence of nNOS in the vessels and parenchymal tissue was observed after 6 mo of treatment, and the presence of iNOS was detected in the vessel walls after 3 mo, in the nerve fibers after 6 mo and in the odontoblasts after 14 mo of treatment. |
| Immunohistochemical and assessment with spectrophotometer Times: before treatment (T1), after 14 mo (T2), after 24 mo (T3) Tissue factors: MMP-2, MMP-9 and iNOS | 1. A reduction of MMP-2 and MMP-9 expression occurred in treated samples after 14 and 24 mo of treatment, P < .05 2. No significant differences were observed in the iNOS levels, P > .05 |
| Immunohistochemical and assessment with spectrophotometer Times: before treatment (T1), after 3 mo (T2), after 6 mo (T3) Tissue factors: HSP60, caspases 3 and 9, and PCNA. | 1. The levels of caspases, PCNA and HSP60 increased after T3, compared to T1 and T2, P < .05 |
| Radioimmunoassay Tissue factors: ME and substance P | 1. The substance P levels were not affected by the application force, P > .05 2. The levels of ME showed response sex-specific: in males the mean concentration decreases (control-group: 20 ± 26.7 pg/mg, ortho group: 12 ±9.4 pg/mg, P < .05) and in females increased (control-group: 26 ± 68.1 pg/mg, ortho group: 139 ± 346.4 pg/mg, P < .05) in response to orthodontic force |
| Immunohistochemical and assessment with spectrophotometer Tissue factors: AST activity | 1. No significant differences in AST levels between the control-group (1.787 ± 1.133 U/mL) and orthodontic group (1.942 ± 1.133), P > .05 |
| Immunohistochemical and assessment with spectrophotometer Tissue factors: AST activity | 1. Significant differences in AST levels between the control-group (3.6 ± 1.4 U/mL) and orthodontic group (6.7 ± 1.9 U/mL), P < .01 |
| Immunohistochemical and assessment with spectrophotometer Tissue factors: AP activity | 1. Significant differences in AP levels between the control-group (142 ± 33 U/mL) and orthodontic-group (89 ± 0.26), P < .01 |
| Immunohistochemical and assessment with spectrophotometer Tissue factors: AST activity | 1. Significant differences in AST levels between the control group (0.35 ± 0.24 U/mL) and orthodontic group (0.57 ± 0.44), P < .01 |
| Immunohistochemical and assessment with spectrophotometer Tissue factors: AST activity | 1. No significant differences in AST levels between T1 (0.27 ± 0.15 U/mL) and T2 (0.21 ± 0.15 U/mL), P = .32 |
| Immunohistochemical and assessment with spectrophotometer Tissue factors: AST activity | 1. No significant differences in AST levels between the control-group (25.29 ± 9.95 U/mL) and orthodontic group (25.54 ± 31.81), P = .312 |
| Reversed phase high-performance liquid chromatography (RP-HPLC), radioreceptor assay (RRA), radioimmunoassay (RIA), and mass spectrometry (MS) Tissue factors: ME | 1. Significant differences in ME levels by RIA analysis between the control-group (43.1 ng/g) and orthodontic-group (20.9 ng/g), P < .05 2. ME concentrations of the teeth extracted decreased in proportion to increasing forces |