Poly(Aspartic Acid) Promotes Odontoblast-like Cell Differentiation in Rat Molars with Exposed Pulp.

In recent years, alternative pulpal therapies targeting dentinogenesis signaling pathways using different peptides have been investigated. The aim of this study was to verify the effectiveness of poly(aspartic acid), pAsp, in dentin regeneration using an animal model. METHODS: Mechanical pulp exposure was performed in the upper molars of 56 Wistar rats, randomly divided as follows (n = 14): control (no treatment); MTA group-pulp capping with mineral trioxide aggregate (MTA Angelus); pAsp group-application of 20 µL of pAsp solution (25 mg·mL-1); MTA+pAsp group-application of MTA mixed with pAsp (5:1 by mass). Animals were euthanized after 7 or 21 days. Histological sections were submitted to hematoxylin-eosin and Brown and Brenn staining and immunohistochemical analysis for osteopontin (OPN) and dentin matrix protein 1 (DMP 1). RESULTS: At 7 days, an acute inflammatory infiltrate and the presence of disorganized mineralized tissue were observed in all groups. At 21 days, the quality and thickness of the reparative dentin in treated groups were superior to the control, and bacterial contamination was observed in two MTA-pAsp specimens. While all treated groups showed intense immunostaining for OPN at 21 days, only the pAsp group expressed DMP 1, indicating the presence of fully differentiated odontoblast-like cells. CONCLUSION: Poly(aspartic) acid promoted dentin regeneration in rat molars in the absence of an additional calcium source and may be an alternative to MTA as a pulp-capping agent.

Temporo-spatial distribution of stem cell markers CD146 and p75NTR during odontogenesis in mice.

Mesenchymal and epithelial stem cells were identified in dental tissues; however, knowledge about the odontogenic stem cells is limited, and there are some questions regarding their temporo-spatial dynamics in tooth development. OBJECTIVE: Our study aimed to analyze the expression of the stem cell markers CD146 and p75NTR during the different stages of odontogenesis. METHODOLOGY: The groups consisted of 13.5, 15.5, 17.5 days old embryos, and 14 days postnatal BALB/c mice. The expression of CD146 and p75NTR was evaluated by immunohistochemistry. RESULTS: Our results showed that positive cells for both markers were present in all stages of tooth development, and the number of positive cells increased with the progression of this process. Cells of epithelial and ectomesenchymal origin were positive for CD146, and the expression of p75NTR was mainly detected in the dental papilla and dental follicle. In the postnatal group, dental pulp cells were positive for CD146, and the reduced enamel epithelium and the oral mucosa epithelium showed immunostaining for p75NTR. CONCLUSIONS: These results suggest that the staining pattern of CD146 and p75NTR underwent temporal and spatial changes during odontogenesis and both markers were expressed by epithelial and mesenchymal cell types, which is relevant due to the significance of the epithelial-ectomesenchymal interactions in tooth development.

Effect of Er:YAG Laser and Association of Protocols on the Demineralized Enamel Microhardness.

Objective: The aim of this study was to analyze the microhardness of demineralized enamel following different treatments (fluoride varnish, Er:YAG laser, and Er:YAG laser associated with fluoride varnish). Methods: Forty-eight enamel blocks (4 × 4 × 7 mm) were divided into six groups (n = 8): (S) Sound; (DE) Demineralized; (DED) DE + Duraphat® 5% (fluoride varnish); (DEL20) DE + Er:YAG laser (20 mJ pulse mode; 0.20 W; 10 Hz; 60 sec; 1.18 J/cm2; 11.83 W/cm2); (DEL50) DE + Er:YAG laser (50 mJ pulse mode; 0.50 W; 10 Hz; 60 sec; 2.95 J/cm2; 29.58 W/cm2); (DEL20D) DE + Er:YAG laser (20 mJ) + Duraphat 5%. The irradiation was performed at 1 mm distance from the surface using a tip (AS7066X, L-14 mm, D-1.3 mm in diameter) in water/air spray refrigeration (level 6). The enamel blocks were submitted to pH cycling (4 h into DES solution +20 h into RE solution for 8 days and the solutions were changed every day). Knoop microhardness was measured (50 g/15 sec, six readings per sample) and data were analyzed by Kruskal-Wallis test at 5% significance. Results: After treatments, DF group showed higher microhardness values than all the groups. Also, DEL20D group showed similar results with H group according to the microhardness analysis (p < 0.05). Conclusions: It could be concluded that Duraphat 5% treatment showed better results when compared with all tested groups, however, the association of Er:YAG Laser 20 with Duraphat 5% also showed promising results.

Development and characterization of a new chitosan-based scaffold associated with gelatin, microparticulate dentin and genipin for endodontic regeneration.

OBJECTIVE: An ideal scaffold for endodontic regeneration should allow the predictableness of the new tissue organization and limit the negative impact of residual bacteria. Therefore, composition and functionalization of the scaffold play an important role in tissue bioengineering. The objective of this study was to assess the morphological, physicochemical, biological and antimicrobial properties of a new solid chitosan-based scaffold associated with gelatin, microparticulate dentin and genipin. METHODS: Scaffolds based on chitosan (Ch); chitosan associated with gelatin and genipin (ChGG); and chitosan associated with gelatin, microparticulate dentin and genipin (ChGDG) were prepared by using the freeze-drying method. The morphology of the scaffolds was analyzed by scanning electron microscopy (SEM). The physicochemical properties were assessed for biodegradation, swelling and total released proteins. The biological aspects of the scaffolds were assessed using human cells from the apical papilla (hCAPs). Cell morphology and adhesion to the scaffolds were evaluated by SEM, cytotoxicity and cell proliferation by MTT reduction-assay. Cell differentiation in scaffolds was assessed by using alizarin red assay. The antimicrobial effect of the scaffolds was evaluated by using the bacterial culture method, and bacterial adhesion to the scaffolds was observed by SEM. RESULTS: All the scaffolds presented porous structures. The ChCDG had more protein release, adhesion, proliferation and differentiation of hCAPs, and bacteriostatic effect on Enterococcus faecalis than Ch and ChGG (p < 0.05). SIGNIFICANCE: The chitosan associated with gelatin, microparticulate dentin and genipin has morphological, physicochemical, biological and antibacterial characteristics suitable for their potential use as scaffold in regenerative endodontics.

In vitro evaluation of hydroxyapatite, chitosan, and carbon nanotube composite biomaterial to support bone healing / Avaliação in vitro do biomaterial compósito à base de hidroxiapatita, quitosana e nanotubo de carbono como adjuvante no reparo ósseo

Hydroxyapatite, chitosan, and carbon nanotube composite biomaterial were developed to improve bone healing. Previous studies suggested that a combination of biomaterials and mesenchymal stem cells (MSCs) can potentially help promote bone regeneration. In the present study, we first developed hydroxyapatite, chitosan, and carbon nanotube composite biomaterial. Then, the effect of different concentrations of the extract on the viability of Vero cells (ATCC CCL-81) and MSCs obtained from sheep bone marrow using methylthiazol tetrazolium (MTT) and propidium iodide (PI) assays were evaluated. The biomaterial group demonstrated an absence of cytotoxicity, similar to the control group. Samples with 50% and 10% biomaterial extract concentrations showed higher cell viability compared to samples from the control group (MTT assay). These results suggest that the presence of this composite biomaterial can be used with MSCs. This study also concluded that hydroxyapatite, chitosan, and carbon nanotube composite biomaterial were not cytotoxic. Therefore, these could be used for performing in vivo tests.(AU)

O compósito à base de hidroxiapatita, quitosana e nanotubo de carbono foi desenvolvido com o intuito de auxiliar na consolidação óssea. Estudos anteriores sugerem que a combinação de substitutos ósseos e células-tronco mesenquimais (CTM) podem auxiliar a potencializar e promover a regeneração óssea. No presente estudo, o biomaterial foi desenvolvido e a viabilidade e a citotoxicidade de células Vero (ATCC CCL-81) e CTM obtidas de medula óssea provenientes de ovinos utilizando ensaios metil-tiazol-tetrazólio, MTT e iodeto de propídeo (PI) foram avaliadas em diferentes concentrações de extrato desse compósito. O compósito demonstrou ausência de citotoxicidade com comportamento semelhante ao grupo controle. Amostras com 50% e 10% de concentração de extrato do compósito mostraram resultados maiores comparados ao grupo controle (ensaio MTT). Esses resultados também sugerem que a presença do biomaterial pode ser utilizada em associação a CTM. Assim, esse estudo conclui que o compósito apresentado de hidroxiapatita, quitosana e nanotubo de cabono não foi considerado citotóxico e pode ser utilizado em teste in vivo.(AU)

Temporo-spatial distribution of stem cell markers CD146 and p75NTR during odontogenesis in mice

Abstract Mesenchymal and epithelial stem cells were identified in dental tissues; however, knowledge about the odontogenic stem cells is limited, and there are some questions regarding their temporo-spatial dynamics in tooth development. Objective Our study aimed to analyze the expression of the stem cell markers CD146 and p75NTR during the different stages of odontogenesis. Methodology The groups consisted of 13.5, 15.5, 17.5 days old embryos, and 14 days postnatal BALB/c mice. The expression of CD146 and p75NTR was evaluated by immunohistochemistry. Results Our results showed that positive cells for both markers were present in all stages of tooth development, and the number of positive cells increased with the progression of this process. Cells of epithelial and ectomesenchymal origin were positive for CD146, and the expression of p75NTR was mainly detected in the dental papilla and dental follicle. In the postnatal group, dental pulp cells were positive for CD146, and the reduced enamel epithelium and the oral mucosa epithelium showed immunostaining for p75NTR. Conclusions These results suggest that the staining pattern of CD146 and p75NTR underwent temporal and spatial changes during odontogenesis and both markers were expressed by epithelial and mesenchymal cell types, which is relevant due to the significance of the epithelial-ectomesenchymal interactions in tooth development.

Mechanical properties and surface roughness of polymer-based materials containing DCPD particles.

The purpose of this study was to synthesize dicalcium phosphate dihydrate (DCPD) particles functionalized with triethylene glycol dimethacrylate (TEGDMA) through different routes by varying the receptor solution: ammonium phosphate (AP groups) or calcium nitrate (CN groups) and the moment in which TEGDMA was incorporated: ab initio (ab) or at the end of dripping the solution (ap). Two syntheses were performed without adding TEGDMA (nf). The particles were characterized by X-ray diffractometry, true density (using a helium pycnometer), surface area, and scanning electron microscopy. A 20 vol% of DCPD particles from the D, E, and F groups was added to the resin matrix to determine the degree of conversion (DC), biaxial flexural strength (BFS), the flexural modulus (FM), and surface roughness after an abrasive challenge (RA). A group with silanized barium glass particles was tested as a control. The data were submitted to ANOVA/Tukey's test (DC, BFS, and RA), and the Kruskal-Wallis test (FM) (alpha = 0.05). BFS values varied between 83 and 142 MPa, and the CN_ab group presented a similar value (123 MPa) to the control group. FM values varied between 3.6 and 8.7 GPa (CN_ab and CN_nf groups, respectively), with a significant difference found only between these groups. RA did not result in significant differences. The use of calcium nitrate solution as a receptor, together with ab initio functionalization formed particles with larger surface areas. Higher BFS values were observed for the material containing DCPD particles with a higher surface area. In general, the DC, FM, and RA values were not affected by the variables studied.

Early effect of laser irradiation in signaling pathways of diabetic rat submandibular salivary glands.

Low-power laser irradiation (LPLI) is clinically used to modulate inflammation, proliferation and apoptosis. However, its molecular mechanisms are still not fully understood. This study aimed to describe the effects of LPLI upon inflammatory, apoptotic and proliferation markers in submandibular salivary glands (SMGs) in an experimental model of chronic disorder, 24h after one time irradiation. Diabetes was induced in rats by the injection of streptozotocin. After 29 days, these animals were treated with LPLI in the SMG area, and euthanized 24h after this irradiation. Treatment with LPLI significantly decreased diabetes-induced high mobility group box 1 (HMGB1) and tumor necrosis factor alpha (TNF-α) expression, while enhancing the activation of the transcriptional factor cAMP response element binding (CREB) protein. LPLI also reduced the expression of bax, a mitochondrial apoptotic marker, favoring the cell survival. These findings suggest that LPLI can hamper the state of chronic inflammation and favor homeostasis in diabetic rats SMGs.

In vitro evaluation of enamel surface roughness and morphology after orthodontic debonding: Traditional cleanup systems versus polymer bur.

INTRODUCTION: The primary aim of this randomized in vitro study was to compare the effectiveness of carbide, fibreglass and polymer burs on resinous remnant removal after bracket debonding, by the evaluation of enamel surface roughness and morphology. The secondary objective was to compare the time dispended on the procedures. METHODS: The buccal surfaces of 28 bovine incisors were analysed by a profilometer to initial roughness measurement (Ra-T1). Brackets were bonded with a light-cured resin and debonded with a debonding plier. The samples were randomly divided into four groups, according to the bur used (n=7): A-Tungsten carbide; B-Fibreglass; C-Polymer; D-Polymer with 75% ethanol pre-treatment. The second roughness measurements were made after resin removal (Ra-T2). Time for removal procedures was also recorded. The third measurements were made after polishing (Ra-T3). Scanning Electronic Microscopy was performed in two samples of each group: after resin removal and after polishing. Results of roughness and time measurements were statically analysed by analysis of variance with post-hoc Bonferroni. RESULTS: After polishing, tungsten carbide (P=0.1555) and fibreglass burs provided final surface roughness statistically similar to the baseline condition (P=1.0000). Yet, polymer burs, associated (P<0.0001) or not to alcohol (P<0.0001), provided surface roughness significantly higher when compared to baseline values. Polymer burs were more time-consuming on resinous remnant removal than tungsten carbide and fibreglass burs (P<0.05). CONCLUSION: Polymer burs were less effective and more time-consuming to remove the remaining resin than tungsten carbide and fibreglass burs. The polishing step created smoother surfaces regardless of the burs used for resin removal.

Improvement of full-thickness rat skin wounds by photobiomodulation therapy (PBMT): A dosimetric study.

Basic dosimetric studies are necessary to support the use of photobiomodulation therapy (PBMT), since the great variety of laser parameters that are reported in the literature have created an obstacle to identifying reproducible results. Thus, the present study evaluates the process of tissue repair after the photobiomodulation therapy, taking into consideration the dose, frequency and the mode of energy delivery used. For this, 6 mm diameter wounds were created on dorsal skin of Wistar rats, and the animals were divided in control and irradiated groups, where L1 and L4 (irradiated with 1 point of 10 J/cm2), L2 and L5 (5 points of 10 J/cm2), L3 and L6 (1 point of 50 J/cm2), respectively for one or multiple days of irradiations. A diode laser, λ 660 nm, 40 mW of power and 0.028 cm2 of spot area was used. Our data showed that the group receiving multiple treatments over the first week post wounding, applied at 10 J/cm2 at each of 5 points on and around the wound (group L5) presented the best improvement of wound closure, higher cytokeratin 10, lower macrophage infiltration, and greater tissue resistance to rupture. We conclude that PBMT improves the skin wound healing process, and the outcomes were directly related to the chosen laser parameters and irradiation mode.

Aquaporin 1, 3, and 5 Patterns in Salivary Gland Mucoepidermoid Carcinoma: Expression in Surgical Specimens and an In Vitro Pilot Study.

Salivary gland aquaporins (AQPs) are essential for the control of saliva production and maintenance of glandular structure. However, little is known of their role in salivary gland neoplasia. Salivary gland tumors comprise a heterogeneous group of lesions, featuring variable histological characteristics and diverse clinical behaviors. Mucoepidermoid carcinoma (MEC) is the most common salivary gland malignancy. The aim of this study was to evaluate the expression of AQP1, AQP3, and AQP5 in 24 MEC samples by immunohistochemistry. AQP1 expression was observed in vascular endothelium throughout the tumor stroma. AQP3 was expressed in epidermoid and mucosal cells and AQP5 was expressed in mucosal cells of MEC. These proteins were expressed in the human MEC cell line UH-HMC-3A. Cellular ultrastructural aspects were analyzed by electron microscopy to certificate the tumor cell phenotype. In summary, our results show that, despite the fact that these molecules are important for salivary gland physiology, they may not play a distinct role in tumorigenesis in MEC. Additionally, the in vitro model may offer new possibilities to further investigate mechanisms of these molecules in tumor biology and their real significance in prognosis and possible target therapies.

Phototherapy With LED as an Effective Treatment for Chemotherapy-Induced Oral Mucositis in Hamsters.

Introduction: Oral mucositis (OM) has been considered one of the most feared collateral effects of oncological treatments. Some therapies have been used, such as light-emitting diode (LED), with promising results, but with no sufficient evidence in the literature. Objective: Our study aimed to evaluate, by clinical and histological analysis, the effect of LED on the treatment of chemotherapy-induced OM (CIOM) in an animal model. Methods: Twenty male hamsters were equally distributed to two groups: control (C), which received anesthesia and CIOM induction; and LED (L), which received anesthesia, CIOM induction, and LED treatment (635 nm, 120 mW, 0.48 J). The clinical analysis was performed through two specific scales for OM analysis on days 5, 7 and 10 of the experiment. In addition, the injured area of all hamsters check pouch mucosa was removed and processed for histological analysis on the last experimental day. Results: After statistical analysis, group L showed less severity of OM when compared with the C group (P <0.05); beyond that, both healed completely on day 10. Conclusion: Our results suggested that the phototherapy with LED had a positive effect on accelerating repair, reducing the severity of CIOM.

Mechanical properties and surface roughness of polymer-based materials containing DCPD particles

Abstract The purpose of this study was to synthesize dicalcium phosphate dihydrate (DCPD) particles functionalized with triethylene glycol dimethacrylate (TEGDMA) through different routes by varying the receptor solution: ammonium phosphate (AP groups) or calcium nitrate (CN groups) and the moment in which TEGDMA was incorporated: ab initio (ab) or at the end of dripping the solution (ap). Two syntheses were performed without adding TEGDMA (nf). The particles were characterized by X-ray diffractometry, true density (using a helium pycnometer), surface area, and scanning electron microscopy. A 20 vol% of DCPD particles from the D, E, and F groups was added to the resin matrix to determine the degree of conversion (DC), biaxial flexural strength (BFS), the flexural modulus (FM), and surface roughness after an abrasive challenge (RA). A group with silanized barium glass particles was tested as a control. The data were submitted to ANOVA/Tukey's test (DC, BFS, and RA), and the Kruskal-Wallis test (FM) (alpha = 0.05). BFS values varied between 83 and 142 MPa, and the CN_ab group presented a similar value (123 MPa) to the control group. FM values varied between 3.6 and 8.7 GPa (CN_ab and CN_nf groups, respectively), with a significant difference found only between these groups. RA did not result in significant differences. The use of calcium nitrate solution as a receptor, together with ab initio functionalization formed particles with larger surface areas. Higher BFS values were observed for the material containing DCPD particles with a higher surface area. In general, the DC, FM, and RA values were not affected by the variables studied.

Effect of the software binning and averaging data during microcomputed tomography image acquisition.

This study describes the effect of the software binning and data averaging during micro CT volume acquisition, on the assessment of root resorption volumes. The mesial roots (n = 9), after orthodontic tooth movement during 14 days, were scanned, using a micro CT system (9 µm/pixel). All roots were reconstructed and the volumes of the resorption lacunae evaluated. The height and width of the pixels vary according to the parameters (A1, A2, A3, A4, A5, A6, A7, A8, A9) used during the scan. In the root #1 the mean volumes of resorption were similar in A4 and A7; in the root #2 there was no similarity in the mean volumes of resorption in any of the parameters; in root #3 only A4 presented mean volume different from zero (3.05 × 10°). In the root #5, the A1 and A7 presented similar mean volumes and in the A6 and A9 presented near mean volumes. In the root #9 the A1, A4, and A7 presented similar mean volumes and A6 and A9 also had similar mean volumes. Significant difference was detected in the volume of resorption among the roots #2, #5 and #9 (p = 0.04). When analyzing delicate structures such as the roots of rats' molars, the variation of such parameters will significantly influence the results.

Infrared light-emitting diode (led) effects on orthodontic tooth movement

Abstract The present study aimed to analyze the effect of LED phototherapy on the presence of hyalinization and root resorption during orthodontic tooth movement (OTM) in rats and to measure the amount of tooth movement. Eighty rats were allocated into two groups: LED and control (CON), where the LED rats were irradiated with infrared LED (850 nm, 30 mW) for 5 min during the first five days of OTM and where controls were not irradiated. Both groups were subdivided into four subgroups (n=10) according to the date of euthanasia (4, 7, 14 and 21 days). Five out of ten LED21 and five of ten CON21 rats were submitted to micro-computed tomography (μCT); μCT scans were taken on days 0, 7, 14 and 21. For histological study, maxillae were processed to light microscopy using Hematoxylin-Eosin (HE) and Tartrate-Resistant Acid Phosphatase (TRAP) histochemistry. The amount of tooth movement did not differ between LED and CON. Hyalinization was observed at the pressure areas in both groups, and it did not show a statistically significant difference between the groups. Root resorption was also observed in both groups after 7 days and it did not represent any differences between the two groups. LED phototherapy was not able to increase the amount of OTM. Similar characteristics of hyalinization and root resorption were observed in both groups.

Resumo O presente estudo tem como objetivo analisar o efeito da fototerapia LED na presença da hialinização e reabsorção radicular durante o movimento dentário ortodôntico (MDO) em ratos, e a mensuração da quantidade de movimento dentário. Oitenta ratos foram alocados em dois grupos: LED e Controle (CON), os ratos foram irradiados com um LED infravermelho (850nm, 30mW) por 5 minutos durante os cinco primeiros dias da MDO; e o grupo controle não foi irradiado. Ambos os grupos foram subdivididos em 4 subgrupos (n=10) de acordo com a data da eutanásia (4, 7, 14 e 21 dias). Cinco dos dez ratos LED21 e cinco dos dez ratos CON21foram submetidos a microtomografia computadorizada (μCT); As μCT foram realizadas nos dias 0, 7, 14 e 21. Para o estudo histológico, as maxilas foram processadas para microscopia de luz, usando hematoxilina-eosina (HE) e Fosfatase ácido Tartrate-Resistente (TRAP) para histoquímica. A quantidade de movimento dentário não diferiu entre o LED e o CON. A hialinização foi observada nas áreas de pressão em ambos os grupos e não mostrou diferença estatisticamente significante. Reabsorção radicular também foi observada em ambos os grupos depois de 7 dias e não houve diferença entre os grupos. A fototerapia LED não aumentou a quantidade de MDO. Características similares de hialinização e reabsorção radicular foram observadas em ambos os grupos

Effect of fluoride application during radiotherapy on enamel demineralization.

OBJECTIVE: Radiation-related caries are one the most undesired reactions manifested during or after head and neck radiotherapy. Fluoride application is an important strategy to reduce demineralization and enhance remineralizaton. To evaluate the effect of the topical application of fluoride during irradiation on dental enamel demineralization. MATERIAL AND METHODS: Thirty molars were randomly divided into three groups: Non-irradiated (NI), Irradiated (I), Irradiated with fluoride (IF). Each group was subdivided according to the presence or absence of pH-cycling (n=5). In the irradiated groups, the teeth received 70 Gy. The enamel's chemical composition was measured using Fourier Transform Infrared Spectrometry (organic matrix/mineral ratio - M/M and relative carbonate content - RCC). Vickers microhardness (VHN) and elastic modulus (E) were evaluated at three depths (surface, middle and deep enamel). Scanning electron microscopy (SEM) was used to assess the enamel's morphology. RESULTS: The FTIR analysis (M/M and RCC) showed significant differences for irradiation, pH-cycling and the interaction between factors (p<0.001). Without pH-cycling, IF had the lowest organic matrix/mineral ratio and relative carbonate content. With pH-cycling, the organic matrix/mineral ratio increased and the relative carbonate content decreased, except for IF. VHN was influenced only by pH-cycling (p<0.001), which generated higher VHN values. ANOVA detected significant differences in E for irradiation (p<0.001), pH-cycling (p<0.001) and for the interaction between irradiation and pH-cycling (p<0.001). Increased E was found for group I without pH-cycling. With pH-cycling, groups I and IF were similar, and showed higher values than NI. The SEM images showed no morphological changes without pH-cycling. With pH-cycling, fluoride helped to maintain the outer enamel's morphology. CONCLUSIONS: Fluoride reduced mineral loss and maintained the outer morphology of irradiated and cycled enamel. However, it was not as effective in preserving the mechanical properties of enamel. Radiotherapy altered the enamel's elastic modulus and its chemical composition.

Pulp analysis of teeth submitted to different types of forces: a histological study in rats.

OBJECTIVE: The purpose of this study was to histologically evaluate pulp and dentin under induced tooth movement (ITM) with different types of forces. MATERIAL AND METHODS: The maxillary right first molars of rats were submitted to movement with continuous (CF), continuous interrupted (CIF) and intermittent (IF) forces during 5, 7 and 9 days with nickel-titanium (NiTi) closed-coil springs exerting 50cN force magnitude. The groups were histologically evaluated as for cellularity pattern, presence of dystrophic, hemodynamic alterations in the pulp as well dentin alterations. The main observed alterations were related to hemodynamic pulp characteristics, such as presence of thrombosis, vascular congestion and hemorrhages. The hemodynamic alterations were statistically evaluated by Shapiro-Wilk normality test and analysis of variance by the Kruskall-Wallis test. RESULTS: There was no significant differences observed between groups in the different types of applied forces and duration of ITM (vascular congestion, p=1.000; hemorrhage, p=0.305; thrombosis, p=1.000). CONCLUSIONS: Pulp tissue alterations resulting from ITM were limited to hemodynamic events, without progressing to irreversible degeneration, regardless of the type of force applied.

Histological and Radiographic Evaluation of Equine Bone Structure after Implantation of Castor Oil Polymer.

OBJECTIVE: The main purpose of this study was to evaluate the characteristics of a vegetal polymeric biomaterial intended for bone substitution in horses and to investigate the responses of the equine third metacarpal bone to biomaterial implantation. MATERIALS AND METHODS: Six horses were submitted to osteotomy on the dorsal aspect of the left and right third metacarpal bones; one bone defect was randomly selected for treatment with biopolymer, while the other was left untreated and served as a control. Bone density was monitored radiographically after surgery and bone biopsy fragments were collected at the end of the 120-day follow-up period. Biopsy fragments were analysed using light and scanning electron microscopy. RESULTS: Mean bone density values (mmAL) were greater in control defects (16.33 ± 1.6) than in polymer-treated defects (14.17 ± 1.7) at 120 days (p = 0.027). Light microscopy revealed greater percentages of new bone formation in control defects (50.15 ± 14.8) than in polymer treated defects (26.94 ± 12.1) at 120 days (p < 0.0001). Scanning electron microscopy analysis suggested a similar quality of pre-existing bone and new bone formed in the presence of biomaterial. CLINICAL SIGNIFICANCE: The absence of adverse reactions supports biomaterial biocompatibility and osteoconducting capacity and suggests the castor oil polymer is a suitable bone substitute for the treatment of bone defects in horses.

Immunohistological study of the effect of vascular Endothelial Growth Factor on the angiogenesis of mature root canals in rat molars.

Tissue bioengineering has been applied to Endodontics to seek a more biological treatment. The presence of blood vessels is crucial for cell nutrition during tissue formation. Objective This study analysed the application of vascular endothelial growth factor (VEGF) in the angiogenesis of mature root canals. Material and methods Upper first molars of twelve 13-week old Wistar male rats were used. The root pulp of the mesiobuccal canal was removed and the root canal instrumented with K-files up to size #25. Periapical bleeding was induced into the root canal by introducing a #15 K-file beyond the apex. The teeth on the right side of the arch were filled up with blood clot (G1), whereas those on the left side were filled up with blood clot plus 50 ng/ml of VEGF (G2). Teeth were sealed with light-curing glass-ionomer cement and the animals were sacrificed after 60 days. The maxilla was dissected and fixed before obtaining serial sections for histological processing with haematoxylin-eosin (HE) and immunohistochemical factor-VIII. Immunohistochemical labelling was evaluated using scores for statistical analysis. Results Immunohistological analysis demonstrated the presence of angiogenesis in both groups, but with higher angiogenic maturation in G2 during the experimental period (p<0.05). HE staining showed connective tissue with absence of odontoblasts in all specimens. Conclusions It can be concluded that it is possible to obtain angiogenesis in mature root canals with or without the use of VEGF, although the latter tends to accelerate blood vessel formation.

Influence of radiotherapy on the dentin properties and bond strength.

OBJECTIVES: The objective of this study was to evaluate the biomechanical properties of dentin and the microtensile bond strength (µTBS) performed before or after radiotherapy (RT). MATERIAL AND METHODS: Dentin chemical composition (infrared spectroscopy-FTIR), SEM images, and mechanical properties (Vickers microhardness-VHN and elastic modulus-E) were evaluated comparing no irradiated and irradiate dentin (n = 5). RT was defined by application of 72 Gy (1.8 Gy daily, 5 days per week, during 8 weeks) with sample immersed in distilled water. µTBS evaluated three groups (n = 10): NI-no irradiated; IB-irradiation before restoration; and IA-irradiation after restoration. Resin-dentin sticks (1.0 mm2) were obtained and submitted to µTBS. Analysis of the bonding interface was made by confocal microscopy. RESULTS: After RT, percentage ratio of FTIR analysis showed increased absorption for all bands. SEM image showed a disorganized dentin structure. Two-way ANOVA showed increased VHN (p = 0.005) and decreased E (p < 0.001). For µTBS, one-way ANOVA and Duncan test showed significant differences among groups (p = 0.018). IB group presented the lowest bond strength values. CONCLUSIONS: RT alters the absorption bands and SEM images showed a disorganization of the dentin structure. Mechanical properties were changed with increased VHN and decreased E. µTBS was affected by the radiotherapy and restoration period (before or after). CLINICAL RELEVANCE: RT causes changes that contribute to increased risk of tooth decay. Restorative treatments can be performed using adhesive procedures, but it is preferable to be performed before of the irradiation protocol, to guarantee better adhesive properties to restoration.