RESUMEN
The purposes of this study are to evaluate the effects of photobiomodulation (PBM) with laser and LED on rat calvaria osteoblasts (rGO lineage), cultured in osteogenic (OST) or regular (REG) medium, after induction of a quiescent state and to test if PBM is capable of osteogenic induction and if there is a sum of effects when combining OST medium with PBM. Before irradiation, the cells were put in a quiescent state (1% FBS) 24 h, when red (AlGaInP-660 nm) and infrared laser (GaAlAs-808 nm) and LED (637 ± 15 nm) were applied. The groups were as follows: red laser (RL3-5 J/cm2, 3 s and RL5-8.3 J/cm2, 5 s, 1.66 W/cm2); infrared laser (IrL3-5 J/cm2, 3 s and IrL5-8.3 J/cm2, 5 s); LED (LED3-3 s and LED5-5 s, 0.02 J/cm2, 0.885 W/cm2); positive (C+, 10% FBS) and negative control (C-, 1% FBS). For alkaline phosphatase (ALP) and mineralization assays, the cells were cultured in REG (DMEM 10% FBS) and OST medium (DMEM 10% FBS, 50 µg/mL ascorbic acid, 10 mM ß-glycerophosphate). Statistical analysis was performed using ANOVA and Tukey's tests (p < 0.05). RL5 and LED5 increased proliferation, in vitro wound closure, ALP, and mineralization in rGO cells (p < 0.05). PBM with red laser and LED induced mineralization by itself, without osteogenic medium, not observed for infrared laser (p < 0.05). A sum of effects was observed in osteogenic medium and PBM by infrared, red laser, and LED (5 s). Red laser and LED increased proliferation, migration, and secretory phases in rGO cells in a dose-dependent manner. PBM with red laser and LED promotes osteogenic induction by itself. PBM with infrared laser and osteogenic medium potentializes mineralization.
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Rayos Láser , Terapia por Luz de Baja Intensidad , Osteoblastos/efectos de la radiación , Osteogénesis/efectos de la radiación , Cráneo/efectos de la radiación , Fosfatasa Alcalina/metabolismo , Animales , Calcificación Fisiológica/efectos de la radiación , Diferenciación Celular/efectos de la radiación , Proliferación Celular/efectos de la radiación , Supervivencia Celular/efectos de la radiación , Células Cultivadas , RatasRESUMEN
BACKGROUND: Calvaria skin has a reduced thickness, and its initial damage produced by irradiation was scarcely reported. We aimed to identify the initial effects of x-ray irradiation in the rat calvaria skin. METHODS: After approval by the Animal Ethical Committee, calvaria skin sections of five Wistar rats per time point were evaluated on days 4, 9, 14, and 25 following a single 15-Gy x-ray irradiation of the head. The control group was composed of five rats and evaluated on day 4. Sections were assessed using hematoxylin-eosin and Masson's trichrome staining for morphology, inflammation, and fibrosis. Fibrosis was also evaluated by the collagen maturation index from Picrosirius red staining and by cell proliferation using the immunohistochemistry, after 5-bromo-2-deoxyuridine intraperitoneal injection. RESULTS: In irradiated rats, we observed a reduction in epithelial cell proliferation (p = 0.004) and in matrix metalloproteinase-9 expression (p < 0.001), an increase in the maturation index, and with a predominance in the type I collagen fibers, on days 9 and 14 (1.19 and 1.17, respectively). A progressive disorganization in the morphology of the collagen fibers at all time points and changes in morphology of the sebaceous gland cells and hair follicle were present until day 14. CONCLUSIONS: The initial damage produced by a single 15-Gy x-ray irradiation to the rat calvaria skin was a change in the normal morphology of collagen fibers to an amorphous aspect, a temporary absence of the sebaceous gland and hair follicles, and without a visible inflammatory process, cell proliferation, or fibrosis process in the dermis.
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Traumatismos por Radiación/patología , Piel/efectos de la radiación , Animales , Proliferación Celular , Masculino , Ratas , Ratas Wistar , Cráneo/efectos de la radiación , Coloración y Etiquetado , Rayos XRESUMEN
Bioactive glasses (BG) are known for their ability to bond to bone tissue. However, in critical situations, even the osteogenic properties of BG may be not enough to induce bone consolidation. Thus, the enrichment of BG with polymers such as Poly (D, L-lactic-co-glycolic) acid (PLGA) and associated to photobiomodulation (PBM) may be a promising strategy to promote bone tissue healing. The aim of the present study was to investigate the in vivo performance of PLGA supplemented BG, associated to PBM therapy, using an experimental model of cranial bone defect in rats. Rats were distributed in 4 different groups (Bioglass, Bioglass/PBM, Bioglas/PLGA and BG/PLGA/PBM). After the surgical procedure to induce cranial bone defects, the pre-set samples were implanted and PBM treatment (low-level laser therapy) started (808 nm, 100 mW, 30 J/cm2). After 2 and 6 weeks, animals were euthanized, and the samples were retrieved for the histopathological, histomorphometric, picrosirius red staining and immunohistochemistry analysis. At 2 weeks post-surgery, it was observed granulation tissue and areas of newly formed bone in all experimental groups. At 6 weeks post-surgery, BG/PLGA (with or without PBM) more mature tissue around the biomaterial particles. Furthermore, there was a higher deposition of collagen for BG/PLGA in comparison with BG/PLGA/PBM, at second time-point. Histomorphometric analysis demonstrated higher values of BM.V/TV for BG compared to BG/PLGA (2 weeks post-surgery) and N.Ob/T.Ar for BG/PLGA compared to BG and BG/PBM (6 weeks post-surgery). This current study concluded that the use of BG/PLGA composites, associated or not to PBM, is a promising strategy for bone tissue engineering.
Asunto(s)
Sustitutos de Huesos/uso terapéutico , Cerámica/uso terapéutico , Fracturas Óseas/terapia , Luz , Ácido Poliglicólico/uso terapéutico , Cráneo/lesiones , Cicatrización de Heridas/efectos de los fármacos , Animales , Sustitutos de Huesos/química , Sustitutos de Huesos/efectos de la radiación , Trasplante Óseo/métodos , Cementación/métodos , Cerámica/química , Terapia Combinada , Masculino , Ensayo de Materiales , Osteogénesis/efectos de los fármacos , Osteogénesis/efectos de la radiación , Fototerapia/métodos , Ácido Poliglicólico/química , Ratas , Ratas Wistar , Cráneo/efectos de los fármacos , Cráneo/efectos de la radiación , Ingeniería de TejidosRESUMEN
Objective: To evaluate the effect of the Er,Cr:YSGG laser on healing of critical-sized calvarial defects (CSDs) in rats submitted to inhalation of cigarette smoke. Background: Smoking has been implicated with the delay in the bone healing after osteotomy procedures, then the use of the Er,Cr:YSGG laser for osteotomy in smokers could be an alternative to the conventional drills. Methods: One hundred animals were randomly allocated into four groups: trephine-the CSDs were made with a trephine drill in healthy rats; Er,Cr:YSGG-the CSDs were made with the Er,Cr:YSGG laser in healthy rats; Trephine-S-the CSDs were made with a trephine drill in rats exposed to cigarette smoke; and Er,Cr:YSGG-S-the CSDs were made with the Er,Cr:YSGG laser in rats exposed to cigarette smoke. The inhalation of cigarette smoke started 7 days before the surgical procedure until euthanasia (immediately, 7, 15, 30, or 60 days after the surgical procedure). A histometric analysis and a histological description were performed to evaluate (1) the residual linear lengths and bone formation in the CSDs; (2) the quality of bone healing. Results: The use of Er,Cr:YSGG laser induces more bone formation compared with the trephine in smokers; however, the closure of the CSD was only superior in the Er,Cr:YSGG-S group compared to the Trephine-S group at the 60-day period. Conclusions: The use of the Er,Cr:YSGG laser stimulated the bone repair process after osteotomy procedures in animals submitted to exposure of inhalation of cigarette smoke.
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Regeneración Ósea/efectos de la radiación , Láseres de Estado Sólido , Cráneo/efectos de la radiación , Fumar , Cicatrización de Heridas/efectos de la radiación , Animales , Exposición por Inhalación , Ratas , Cráneo/cirugíaRESUMEN
OBJECTIVE: Considering the global public health problem of smoking, which can negatively influence bone tissue repair, the aim of this study is to analyze the influence of photobiomodulation therapy (PBM) on calvaria defects created surgically in specimens under the effect of cigarette smoke and analyzed with use of histomorphometric and immunohistochemistry techniques. METHODOLOGY: Calvaria defects 4.1 mm in diameter were surgically created in the calvaria of 90-day-old rats (n=60) that were randomly divided into 4 experimental groups containing 15 animals each: control group (C), smoking group (S), laser group (L), and smoke associated with laser group (S+L). The animals were subjected to surgery for calvaria defects and underwent PBM, being evaluated at 21, 45, and 60 days post-surgery. The specimens were then processed for histomorphometric and immunohistochemistry analyses. The area of bone neoformation (ABN), percentage of bone neoformation (PBNF), and the remaining distance between the edges of the defects (D) were analyzed histometrically. Quantitative analysis of the TRAP immunolabeled cells was also performed. The data were subjected to analysis of variance (ANOVA) in conjunction with Tukey's test to verify the statistical differences between groups (p<0.05). RESULTS: The smoking group showed less ABN compared to the other experimental groups in all periods, and it also showed more D at 21 days compared to the remaining groups and at 45 days compared to the laser group. The smoking group showed a lower PNBF compared to the laser group in all experimental periods and compared to smoking combined with LLLT group at 21 days. CONCLUSIONS: PBM acted on bone biomodulation, thus stimulating new bone formation and compensating for the negative factor of smoking, which can be used as a supportive therapy during bone repair processes.
Asunto(s)
Regeneración Ósea/efectos de los fármacos , Fumar Cigarrillos/fisiopatología , Terapia por Luz de Baja Intensidad/métodos , Cráneo/efectos de la radiación , Cicatrización de Heridas/efectos de la radiación , Animales , Fumar Cigarrillos/efectos adversos , Femenino , Inmunohistoquímica , Osteogénesis/efectos de los fármacos , Distribución Aleatoria , Ratas Wistar , Reproducibilidad de los Resultados , Cráneo/patología , Factores de Tiempo , Resultado del TratamientoRESUMEN
Abstract Objective Considering the global public health problem of smoking, which can negatively influence bone tissue repair, the aim of this study is to analyze the influence of photobiomodulation therapy (PBM) on calvaria defects created surgically in specimens under the effect of cigarette smoke and analyzed with use of histomorphometric and immunohistochemistry techniques. Methodology Calvaria defects 4.1 mm in diameter were surgically created in the calvaria of 90-day-old rats (n=60) that were randomly divided into 4 experimental groups containing 15 animals each: control group (C), smoking group (S), laser group (L), and smoke associated with laser group (S+L). The animals were subjected to surgery for calvaria defects and underwent PBM, being evaluated at 21, 45, and 60 days post-surgery. The specimens were then processed for histomorphometric and immunohistochemistry analyses. The area of bone neoformation (ABN), percentage of bone neoformation (PBNF), and the remaining distance between the edges of the defects (D) were analyzed histometrically. Quantitative analysis of the TRAP immunolabeled cells was also performed. The data were subjected to analysis of variance (ANOVA) in conjunction with Tukey's test to verify the statistical differences between groups (p<0.05). Results The smoking group showed less ABN compared to the other experimental groups in all periods, and it also showed more D at 21 days compared to the remaining groups and at 45 days compared to the laser group. The smoking group showed a lower PNBF compared to the laser group in all experimental periods and compared to smoking combined with LLLT group at 21 days. Conclusions PBM acted on bone biomodulation, thus stimulating new bone formation and compensating for the negative factor of smoking, which can be used as a supportive therapy during bone repair processes.
Asunto(s)
Animales , Femenino , Ratas , Cráneo/efectos de la radiación , Cicatrización de Heridas/efectos de la radiación , Regeneración Ósea/efectos de los fármacos , Terapia por Luz de Baja Intensidad/métodos , Fumar Cigarrillos/fisiopatología , Osteogénesis/efectos de los fármacos , Cráneo/patología , Factores de Tiempo , Inmunohistoquímica , Distribución Aleatoria , Reproducibilidad de los Resultados , Resultado del Tratamiento , Ratas Wistar , Fumar Cigarrillos/efectos adversosRESUMEN
Biocompatible membranes are widely used in medicine to stimulate bone repair. Several studies have demonstrated that laser photobiomodulation (PBM) also stimulates osteoblast proliferation and osteogenesis at the fracture site, leading to a greater deposition of bone mass and accelerating the process of bone consolidation. This work assessed the therapeutic effect of 780-nm laser PBM and a polystyrene membrane coated with norbixin and collagen (PSNC) on bone healing in rats with calvarial bone defect. Histological staining, Raman spectroscopy, and scanning electron microscopy (SEM) were used to evaluate the bone repair process. Four experimental treatment groups were compared: C, control; M, membrane only; L, laser PBM only; and ML, membrane + laser PBM. A bone defect was created in the calvaria of each animal, with each group subdivided into two subgroups that underwent euthanasia after 15 and 30 days treatment. The L and ML groups were irradiated (λ = 780 nm, ED = 6 J/cm2, P = 60 mW, t = 4 s) postoperatively on alternate days until they were euthanized. The bone concentration of hydroxyapatite (CHA) showed a clear gradation with increasing phosphate area in the order B (normal cortical bone) > L > M > ML > C for both periods. The PSNC membrane was effective in reducing the inflammatory process and served as a scaffold for bone repair. The laser PBM also showed positive effects on the bone repair process with increased deposition and organization of the newly formed bone. However, laser PBM failed to improve the bioactive properties of the membrane scaffold.
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Huesos/patología , Huesos/efectos de la radiación , Carotenoides/farmacología , Terapia por Luz de Baja Intensidad/métodos , Poliestirenos/química , Cicatrización de Heridas/efectos de los fármacos , Cicatrización de Heridas/efectos de la radiación , Animales , Huesos/efectos de los fármacos , Colágeno/farmacología , Durapatita , Masculino , Microscopía Electrónica de Rastreo , Ratas Wistar , Cráneo/efectos de los fármacos , Cráneo/patología , Cráneo/efectos de la radiación , Espectrometría RamanRESUMEN
The repair of bone defects raises the interest of investigators in several health specialties. Grafting techniques with bone substitutes and laser therapies have been investigated to replace autogenous bone and accelerate the bone healing process. Objective To evaluate the effect of photobiomodulation therapy (PBMT) associated with guided bone regeneration (GBR) in critical size defects. Material and Methods The study was conducted on 80 male rats (Rattus norvegicus albinus, Wistar) submitted to surgical creation of a critical size defect on the calvaria, divided into eight study groups: group C (control - only blood clot); group M (collagen membrane); group PBMT (photobiomodulation therapy); group AB (autogenous bone); group AB+PBMT; group AB+M; group PBMT+M; group AB+PBMT+M. The animals were killed 30 days postoperatively. After tissue processing, bone regeneration was evaluated by histomorphometric analysis and statistical analyses were performed (Tukey test, p<0.05). Results All groups had greater area of newly formed bone compared to group C (9.96±4.49%). The group PBMT+M (achieved the greater quantity of new bone (64.09±7.62%), followed by groups PBMT (47.67±8.66%), M (47.43±15.73%), AB+PBMT (39.15±16.72%) and AB+PBMT+M (35.82±7.68%). After group C, the groups AB (25.10±16.59%) and AB+M (22.72±13.83%) had the smallest quantities of newly formed bone. The area of remaining particles did not have statistically significant difference between groups AB+M (14.93±8.92%) and AB+PBMT+M (14.76±6.58%). Conclusion The PBMT utilization may be effective for bone repair, when associated with bone regeneration techniques.
Asunto(s)
Regeneración Ósea/efectos de la radiación , Regeneración Tisular Dirigida/métodos , Terapia por Luz de Baja Intensidad/métodos , Animales , Autoinjertos , Regeneración Ósea/fisiología , Colágeno/análisis , Masculino , Osteogénesis/fisiología , Osteogénesis/efectos de la radiación , Distribución Aleatoria , Ratas Wistar , Valores de Referencia , Reproducibilidad de los Resultados , Cráneo/fisiología , Cráneo/efectos de la radiación , Cráneo/cirugía , Resultado del Tratamiento , Cicatrización de Heridas/fisiología , Cicatrización de Heridas/efectos de la radiaciónRESUMEN
Abstract The repair of bone defects raises the interest of investigators in several health specialties. Grafting techniques with bone substitutes and laser therapies have been investigated to replace autogenous bone and accelerate the bone healing process. Objective To evaluate the effect of photobiomodulation therapy (PBMT) associated with guided bone regeneration (GBR) in critical size defects. Material and Methods The study was conducted on 80 male rats (Rattus norvegicus albinus, Wistar) submitted to surgical creation of a critical size defect on the calvaria, divided into eight study groups: group C (control - only blood clot); group M (collagen membrane); group PBMT (photobiomodulation therapy); group AB (autogenous bone); group AB+PBMT; group AB+M; group PBMT+M; group AB+PBMT+M. The animals were killed 30 days postoperatively. After tissue processing, bone regeneration was evaluated by histomorphometric analysis and statistical analyses were performed (Tukey test, p<0.05). Results All groups had greater area of newly formed bone compared to group C (9.96±4.49%). The group PBMT+M (achieved the greater quantity of new bone (64.09±7.62%), followed by groups PBMT (47.67±8.66%), M (47.43±15.73%), AB+PBMT (39.15±16.72%) and AB+PBMT+M (35.82±7.68%). After group C, the groups AB (25.10±16.59%) and AB+M (22.72±13.83%) had the smallest quantities of newly formed bone. The area of remaining particles did not have statistically significant difference between groups AB+M (14.93±8.92%) and AB+PBMT+M (14.76±6.58%). Conclusion The PBMT utilization may be effective for bone repair, when associated with bone regeneration techniques.
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Animales , Masculino , Regeneración Ósea/efectos de la radiación , Terapia por Luz de Baja Intensidad/métodos , Regeneración Tisular Dirigida/métodos , Osteogénesis/efectos de los fármacos , Osteogénesis/fisiología , Valores de Referencia , Cráneo/cirugía , Cráneo/efectos de la radiación , Cráneo/fisiología , Cicatrización de Heridas/efectos de la radiación , Cicatrización de Heridas/fisiología , Regeneración Ósea/fisiología , Distribución Aleatoria , Reproducibilidad de los Resultados , Colágeno/análisis , Resultado del Tratamiento , Ratas Wistar , AutoinjertosRESUMEN
The aim of this study was to evaluate the effect of collagen sponge scaffold (CSS) implantation associated with low-level laser therapy (LLLT) on repairing bone defects. A single 5-mm cranial defect was surgically created in forty Wistar rats, which then received one of the following four interventions (n = 10 per group): no treatment (G0); bone defect implanted with collagen sponge scaffold (CSS) alone (G1); defect treated with low-level laser therapy (LLLT) (wavelength 780 nm; total energy density 120 J/cm2 ; power 50 mW) alone (G2); and CSS associated with LLLT treatment (G3). After surgery, animals in each group were euthanized at 21 days and 30 days (n = 5 per euthanasia time group). Bone formation was monitored by X-ray imaging analysis. Biopsies were collected and processed for histological analysis and immunohistochemical evaluation of transforming growth factor-beta (TGF-ß), fibroblast growth factor-2 (FGF-2), osteoprotegerin (OPG) and receptor activator of nuclear factor Æ (RANK). Osteocalcin (OCN) was detected by immunofluorescence analysis. Compared to the G0 group, defects in the 30-day G3 group exhibited increased bone formation, both by increase in radiopaque areas (P < 0.01) and by histomorphometric analysis (P < 0.001). The histopathological analysis showed a decreased number of inflammatory cells (P < 0.001). The combined CCS + LLLT (G3) treatment also resulted in the most intense immunostaining for OPG, RANK, FGF-2 and TGF-ß, and the most intense and diffuse OCN immunofluorescent labelling at 30 days postsurgery (G3 vs. G0 group, P < 0.05). Therefore, the use of CCS associated with LLLT could offer a synergistic advantage in improving the healing of bone fractures.
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Regeneración Ósea/fisiología , Colágeno/uso terapéutico , Terapia por Luz de Baja Intensidad , Osteocalcina/metabolismo , Cráneo/cirugía , Animales , Regeneración Ósea/efectos de la radiación , Factor 2 de Crecimiento de Fibroblastos/metabolismo , Técnica del Anticuerpo Fluorescente , Inmunohistoquímica , Masculino , Microscopía Confocal , Osteocalcina/análisis , Osteoprotegerina/metabolismo , Radiografía , Distribución Aleatoria , Ratas , Ratas Wistar , Método Simple Ciego , Cráneo/diagnóstico por imagen , Cráneo/patología , Cráneo/efectos de la radiación , Factor de Crecimiento Transformador beta/metabolismoRESUMEN
Low-level laser therapy (LLLT) benefits bone metabolism, but its use needs to be standardized. We evaluated the effects of LLLT on bone defects in calvaria of ovariectomized rats. Stereology was used to calculate tissue repair volume (V tr ), density of trabecular bone volume (Vv t ), total volume of newly formed trabecular bone (Vtot), and the area occupied by collagen fibers (A C ). Fifty-four Wistar rats were submitted to bilateral ovariectomy, and bone defects were created in calvaria after 150 days. The animals were divided into nine groups (n = 6), and 24 h after defects, the treatment started with a 780-nm low-intensity GaAlAs laser: G1, G2, and G3 received 3 sessions of 0, 20, and 30 J/cm(2) respectively; G4, G5, and G6 received 6 sessions of 0, 20, and 30 J/cm(2), respectively; and G7, G8, and G9 received 12 sessions of 0, 20, and 30 J/cm(2), respectively. A normal distribution was found for all of the data. The test used to verify the normality was the Kolmogorov-Smirnov (KS, p > 0.05). The one-way ANOVA followed by Tukey's post hoc test was used for data processing. A difference of p < 0.05 was considered statistically significant. Groups G2 and G1 showed significance for V tr , Vv t , Vtot, and (A C ). Results were significant for (Vv t ) and (Vtot) between G3 and G1. There were no significant results between G5 and G4 as well as between G8 and G7. Groups G6 and G4 results showed statistical difference for V tr , Vv t , Vtot, and (A C ). Groups G9 and G7 showed significance for V tr , Vv t , Vtot, and (A C ). In conclusion, there was new bone formation in the groups that received 20 and 30 J/cm(2) when compared to control groups, but over time, the dose of 30 J/cm(2) showed better stereological parameters when compared to 20 J/cm(2).
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Terapia por Luz de Baja Intensidad , Osteogénesis/efectos de la radiación , Osteoporosis/radioterapia , Animales , Femenino , Humanos , Ratas Wistar , Cráneo/patología , Cráneo/fisiopatología , Cráneo/efectos de la radiaciónRESUMEN
OBJECTIVE: The objective of this study was to evaluate the effect of low-level laser (LLL) on bone healing process in surgically created critical size defects in rat calvaria treated with or without autogenous bone graft (AB). MATERIAL AND METHODS: The study was conducted on 40 male rats (Rattusnorvegicus, albinus, Wistar), weighing 250-300 g. For accomplishment of the experimental procedures, the rats were anesthetized with an intramuscular injection of xylazine (0.02 ml/kg) and ketamine hydrochloride (0.4 ml/kg). Acritical size defect with 5-mm diameter was created. The animals were divided into four groups: Group C (Control- filled with blood clot), Group LLL, Group AB (autogenous bone graft), Group AB + LLL (autogenous bone graft and LLL). The animals treated with LLL received applications of LLL at the infrared spectrum wavelength (λ = 810 nm) and energy density of 6 J/cm(2) per point, 60 s per point, adding up to five points on the entire created defect. The animals were euthanized at 30 days postoperatively. After decalcification, each specimen was longitudinally divided into two blocks, exactly along the center of the original surgical defect, processed and embedded in paraffin. Longitudinal serial sections with 6-µm thickness were made, initiating from the center of the original surgical defect. The sections were stained with hematoxylin and eosin (HE) for light microscopy analysis for histomorphometric analysis. RESULTS: Group C presented smaller quantity of new bone formation than Groups LLL (P < 0.01), AB (P < 0.01), and AB + LLL (P < 0.01). CONCLUSIONS: Utilization of LLL favored the healing process in rat calvaria. The quantity of new bone formation with use of the LLL was similar to the autogenous bone graft.
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Trasplante Óseo , Terapia por Luz de Baja Intensidad , Osteogénesis/efectos de la radiación , Cráneo/efectos de la radiación , Cicatrización de Heridas/efectos de la radiación , Animales , Masculino , Ratas , Ratas Wistar , Cráneo/cirugía , Trasplante AutólogoRESUMEN
This study evaluated the osteogenic capacity of a new fibrin sealant (FS) combined with bone graft and laser irradiation in the bone repair. Defects were created in the skull of 30 rats and filled with autogenous graft and FS derived from snake venom. Immediately after implantation, low-power laser was applied on the surgical site. The animals were divided in: control group with autogenous graft (G1), autogenous graft and laser 5 J/cm2 (G2), autogenous graft and laser 7 J/cm2 (G3), autogenous graft and FS (G4), autogenous graft, FS and laser 5 J/cm2 (G5), autogenous graft, FS and laser 7 J/cm2 (G6). The animals were sacrificed 6 weeks after implant. Results showed absence of inflammatory infiltrate in the bone defect. New bone formation occurred in all groups, but it was most intense in G6. Thus, the FS and laser 7 J/cm2 showed osteoconductive capacity and can be an interesting resource to be applied in surgery of bone reconstruction.
Este estudo avaliou a capacidade osteogênica de um novo selante de fibrina (FS) associado com enxerto ósseo e irradiação laser no reparo ósseo. Defeitos foram criados no crânio de 30 ratos e preenchidos com enxerto autógeno e FS derivado do veneno de cobra. Imediatamente após implantação, foi aplicado laser de baixa potência na área cirúrgica. Os animais foram divididos em grupo controle com autógeno (G1), autógeno e laser 5 J/cm2 (G2), autógeno e laser 7J/cm2 (G3), autógeno e FS (G4), autógeno, FS e laser 5J/cm2 (G5), autógeno, FS e laser 7J/cm2 (G6). Os animais foram sacrificados 6 semanas após implante. Resultados mostraram ausência de infiltrado inflamatório no defeito ósseo. Neoformação óssea ocorreu em todos os grupos, entretanto, foi mais intenso em G6. Desta maneira, o FS e laser 7J/cm2 mostraram capacidade osteocondutiva e podem ser um interessante recurso a ser aplicado nas cirurgias de reconstrução óssea.
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Animales , Masculino , Ratas , Adhesivo de Tejido de Fibrina/farmacología , Cráneo/efectos de la radiación , Desarrollo Óseo/efectos de los fármacos , Desarrollo Óseo/efectos de la radiación , Rayos Láser , Ratas Wistar , Cráneo/efectos de los fármacosRESUMEN
This study evaluated the osteogenic capacity of a new fibrin sealant (FS) combined with bone graft and laser irradiation in the bone repair. Defects were created in the skull of 30 rats and filled with autogenous graft and FS derived from snake venom. Immediately after implantation, low-power laser was applied on the surgical site. The animals were divided in: control group with autogenous graft (G1), autogenous graft and laser 5 J/cm2 (G2), autogenous graft and laser 7 J/cm2 (G3), autogenous graft and FS (G4), autogenous graft, FS and laser 5 J/cm2 (G5), autogenous graft, FS and laser 7 J/cm2 (G6). The animals were sacrificed 6 weeks after implant. Results showed absence of inflammatory infiltrate in the bone defect. New bone formation occurred in all groups, but it was most intense in G6. Thus, the FS and laser 7 J/cm2 showed osteoconductive capacity and can be an interesting resource to be applied in surgery of bone reconstruction.
Asunto(s)
Adhesivo de Tejido de Fibrina/farmacología , Cráneo/efectos de la radiación , Animales , Desarrollo Óseo/efectos de los fármacos , Desarrollo Óseo/efectos de la radiación , Rayos Láser , Masculino , Ratas , Ratas Wistar , Cráneo/efectos de los fármacosRESUMEN
The purpose of this study was to analyze histologically the effect of low-level laser therapy (LLLT) in combination with bisphosphonate on bone healing in surgically created critical size defects (CSD) in rat calvaria. One hundred Wistar female rats sham operated (sham) and ovariectomized (Ovx) were maintained untreated for 1 month to allow for the development of osteopenia in the Ovx animals. A CSD was made in the calvarium of each rat, and the animals were divided into five groups according to following treatments: (1) sham rats (control), (2) Ovx rats, (3) Ovx rats treated with LLLT, (4) Ovx rats treated with bisphosphonate, and (5) Ovx rats treated with bisphosphonate and LLLT. Groups 4 and 5 were irrigated with 1 ml of bisphosphonate, and groups 3 and 5 were submitted to LLLT (GaAlAs), 660 nm, 24 J, and 0.4285 W/cm(2) on the CSD. Ten animals of each treatment were killed at 30 and 60 days. Histomorphometric assessments, using image analysis software, and histological analyses were performed. No defect was completely regenerated with the bone. Histometrically, it can be observed that groups 3 (37.49 ± 1.94%, 43.11 ± 2.39%) and 5 (35.05 ± 1.57%, 41.07 ± 1.89%) showed a significant bone neoformation when compared to groups 1 (16.81 ± 1.57%, 27.54 ± 1.49%), 2 (11.68 ± 0.98%, 22.51 ± 1.05%), and 4 (14.62 ± 1.70%, 25.67 ± 1.41%) in all experimental periods (P < 0.05). It was possible to conclude that the LLLT associated or not with bisphosphonate treatment was effective for stimulating bone formation in CSD in the calvaria of rats submitted to ovariectomy.
Asunto(s)
Regeneración Ósea/efectos de los fármacos , Regeneración Ósea/efectos de la radiación , Difosfonatos/farmacología , Terapia por Luz de Baja Intensidad/métodos , Cráneo/efectos de la radiación , Animales , Conservadores de la Densidad Ósea/farmacología , Enfermedades Óseas Metabólicas/cirugía , Interpretación Estadística de Datos , Femenino , Procesamiento de Imagen Asistido por Computador , Ovariectomía , Ratas , Ratas Wistar , Cráneo/efectos de los fármacos , Cráneo/patología , Cráneo/cirugíaRESUMEN
OBJECTIVE: To assess the effect of low-level laser therapy (LLLT) on the incorporation of deep-frozen block allografts in a rabbit model. BACKGROUND DATA: Studies have shown that LLLT has beneficial effects on tissue repair and new bone formation. METHODS: Bone tissue was harvested from two rabbits, processed by deep-freezing and grafted into the calvaria of 12 animals, which were then randomly allocated into two groups: experimental (L) and control (C). Rabbits in group L were irradiated with an aluminum gallium arsenide diode laser (AlGaAs; wavelength 830 nm, 4 J/cm(2)), applied to four sites on the calvaria, for a total dose of 16 J/cm(2) per session. The total treatment dose after eight sessions was 128 J/cm(2). Animals were euthanized at 35 (n = 6) or 70 days (n = 6) postoperatively. RESULTS: Deep-freeze-processed block allografts followed by LLLT showed incorporation at the graft-host interface, moderate bone remodeling, partial filling of osteocyte lacunae, less inflammatory infiltrate in the early postoperative period, and higher collagen deposition than the control group. CONCLUSION: Optical microscopy and scanning electron microscopy showed that allograft bone processed by deep-freezing plus LLLT is suitable as an alternative for the treatment of bone defects. Use of the deep-freezing method for processing of bone grafts preserves the structural and osteoconductive characteristics of bone tissue.
Asunto(s)
Terapia por Luz de Baja Intensidad , Osteogénesis/efectos de la radiación , Trasplante Homólogo , Animales , Trasplante Óseo , Humanos , Masculino , Microscopía Electrónica de Rastreo , Conejos , Cráneo/crecimiento & desarrollo , Cráneo/efectos de la radiaciónRESUMEN
The purpose of this study was to investigate the histological changes that occur in rat soft and hard tissues after Er,Cr:YSGG laser surgery. Each of 20 rats was submitted to four procedures which were randomly distributed to the right and left sides of the animal: procedure 1 dorsal incision with a scalpel; procedure 2 dorsal incision with a 2.0-W Er,Cr:YSGG laser; procedure 3 skull defect created with a diamond bur; procedure 4 skull defect created with a 3.0-W Er,Cr:YSGG laser. The animals were killed 3, 7, 15 and 30 days after surgery, and histological examinations were performed. The histometric analysis of the bone defects was evaluated using an unpaired t-test. Initially, the dorsum showed more histological signs of repair following procedure 1, although similar healing responses following procedures 1 and 2 were seen on day 30 after surgery. By day 30 the bone formation observed following procedure 4 was much more evident than following procedure 3. The unpaired t-test identified significant differences in bone formation on day 30 (p = 0.01), whereas a greater bone percentage was seen following procedure 4 than following procedure 3 (79.96 ± 10.30% and 58.23 ± 9.99%, respectively). Thus, histological repair of the Er,Cr:YSGG laser wounds was similar to that of the scalpel wounds. However, skull defects created with the Er,Cr:YSGG laser showed greater bone formation than defects created with the bur. Within the limitations of this study, we can conclude that the Er,Cr:YSGG laser is a promising surgical instrument in vivo, particularly for bone surgery.
Asunto(s)
Huesos/cirugía , Terapia por Láser/instrumentación , Cráneo/cirugía , Animales , Láseres de Estado Sólido , Masculino , Periodo Posoperatorio , Ratas , Cráneo/efectos de la radiación , Cicatrización de Heridas/efectos de la radiaciónRESUMEN
Low-level laser irradiation (LLLI) and recombinant human bone morphogenetic protein type 2 (rhBMP-2) have been used to stimulate bone formation. LLLI stimulates proliferation of osteoblast precursor cells and cell differentiation and rhBMP-2 recruits osteoprogenitor cells to the bone healing area. This in vivo study evaluated the effects of LLLI and rhBMP-2 on the bone healing process in rats. Critical bone defects were created in the parietal bone in 42 animals, and the animals were divided into six treatment groups: (1) laser, (2) 7 µg of rhBMP-2, (3) laser and 7 µg of rhBMP-2, (4) 7 µg of rhBMP-2/monoolein gel, (5) laser and 7 µg rhBMP-2/monoolein gel, and (6) critical bone defect controls. A gallium-aluminum-arsenide diode laser was used (wavelength 780 nm, output power 60 mW, beam area 0.04 cm(2), irradiation time 80 s, energy density 120 J/cm(2), irradiance 1.5 W/cm(2)). After 15 days, the calvarial tissues were removed for histomorphometric analysis. Group 3 defects showed higher amounts of newly formed bone (37.89%) than the defects of all the other groups (P < 0.05). The amounts of new bone in defects of groups 1 and 4 were not significantly different from each other (24.00% and 24.75%, respectively), but were significantly different from the amounts in the other groups (P < 0.05). The amounts of new bone in the defects of groups 2 and 5 were not significantly different from each other (31.42% and 31.96%, respectively), but were significantly different from the amounts in the other groups (P < 0.05). Group 6 defects had 14.10% new bone formation, and this was significantly different from the amounts in the other groups (P < 0.05). It can be concluded that LLLI administered during surgery effectively accelerated healing of critical bone defects filled with pure rhBMP-2, achieving a better result than LLLI alone or the use of rhBMP-2 alone.
Asunto(s)
Proteína Morfogenética Ósea 2/administración & dosificación , Regeneración Ósea/efectos de los fármacos , Regeneración Ósea/efectos de la radiación , Terapia por Luz de Baja Intensidad , Animales , Femenino , Humanos , Láseres de Semiconductores/uso terapéutico , Ratas , Ratas Wistar , Proteínas Recombinantes/administración & dosificación , Cráneo/efectos de los fármacos , Cráneo/lesiones , Cráneo/patología , Cráneo/efectos de la radiaciónRESUMEN
OBJECTIVE: The aim of this study was to investigate the effects of 670-nm, 780-nm, and 830-nm laser irradiation on cell proliferation of normal primary osteoblast (MC3T3) and malignant osteosarcoma (MG63) cell lines in vitro. BACKGROUND: Some studies have shown that laser phototherapy is able to stimulate the osteogenesis of bone tissue, increasing osteoblast proliferation and accelerating fracture consolidation. It has been suggested that laser light may have a biostimulatory effect on tumor cells. However, the mechanism by which the laser acts on cells is not fully understood. MATERIALS AND METHODS: Neonatal, murine, calvarial, osteoblastic, and human osteosarcoma cell lines were studied. A single laser irradiation was performed at three different wavelengths, at the energies of 0.5, 1, 5, and 10 J/cm(2). Twenty-four hours after laser irradiation, cell proliferation and alkaline phosphatase assays were assessed. RESULTS: Osteoblast proliferation increased significantly after 830-nm laser irradiation (at 10 J/cm(2)) but decreased after 780-nm laser irradiation (at 1, 5, and 10 J/cm(2)). Osteosarcoma cell proliferation increased significantly after 670-nm (at 5 J/cm(2)) and 780-nm laser irradiation (at 1, 5, and 10 J/cm(2)), but not after 830-nm laser irradiation. Alkaline phosphatase (ALP) activity in the osteoblast line was increased after 830-nm laser irradiation at 10 J/cm(2), whereas ALP activity in the osteosarcoma line was not altered, regardless of laser wavelength or intensity. CONCLUSION: Based on the conditions of this study, we conclude that each cell line responds differently to specific wavelength and dose combinations. Further investigations are required to investigate the physiological mechanisms responsible for the contrasting outcomes obtained when using laser irradiation on cultured normal and malignant bone cells.
Asunto(s)
Diferenciación Celular/efectos de la radiación , Proliferación Celular/efectos de la radiación , Terapia por Luz de Baja Intensidad/métodos , Osteoblastos/efectos de la radiación , Osteogénesis/efectos de la radiación , Osteosarcoma/radioterapia , Fosfatasa Alcalina/metabolismo , Análisis de Varianza , Animales , Biomarcadores/metabolismo , Línea Celular , Transformación Celular Neoplásica , Humanos , Técnicas In Vitro , Ratones , Osteoblastos/metabolismo , Osteosarcoma/patología , Cráneo/efectos de la radiación , Células Tumorales CultivadasRESUMEN
La etiología de los meningiomas no está dilucidada. Las radiaciones son uno de los pocos factores etiológicos estudiados que pueden inducir el desarrollo de estos tumores, al provocar alteraciones en el brazo largo del cromosoma 22. Se presenta el caso de una paciente que a raíz de una osteomielitis crónica de la calota, fue irradiada en el cráneo, desarrollando 20 años más tarde múltiples meningiomas de la convexidad, que fueron resecados. Se comentan las características de los meningiomas inducidos por radiaciones, como son la multiplicidad, frecuente localización en la convexidad, tendencia a la malignización y recurrencia precoz, características que se deben tener en cuenta en el tratamiento