Fibrin Network and Platelets Densities in Platelet-Rich Fibrin (PRF) Membranes Produced from Plastic Tubes Without Additives: A New Approach to PRF Clinical Use.

Background/Purpose: The present study aimed to investigate plastic tubes without additives as alternatives to glass and silica-coated plastic tubes, in the production of PRF membranes. Materials and Methods: Nine blood samples were collected from eight volunteers (n = 8) separated into three groups, according to tube material: glass, silica-coated plastic, and plastic without additives. In each group, the samples were centrifuged using different relative centrifugation forces: L-PRF (700 g/12 min), A-PRF (200 g/14 min), and A-PRF + (200 g/8 min). The generated membranes were evaluated by histomorphometry, considering the fibrin network, platelet aggregates, and cellular morphology, by light microscopy. The ultrastructural cellular morphology integrity was evaluated by transmission electron microscopy. Results: The L-PRF (p < 0.019) and A-PRF (p < 0.001) membranes showed a significantly lower fibrin network density in plastic tubes without additives compared to glass and silica-coated plastic tubes. Plastic tubes without additives revealed a significantly higher platelet percentage, regardless of the protocol (p < 0.005). In all groups, TEM analysis showed preserved normal morphological ultrastructure, maintaining the integrity of cellular components. Conclusion: Plastic tubes without additives offer a viable alternative for producing PRF membranes. They exhibited a higher platelet density and demonstrated fibrin network and cellular morphology similar to those of glass and silica-coated plastic tubes, irrespective of the centrifugation protocol.

Insulin Therapy on Bone Macroscopic, Microarchitecture, and Mechanical Properties of Tibia in Diabetic Rats.

BACKGROUND: This study evaluated tibia's macroscopic structure, mechanical properties, and bone microarchitecture in rats with type 1 diabetes mellitus (T1DM). METHODS: Eighteen animals were divided into three groups (n=6): non-diabetic (ND), diabetic (D), and diabetic+insulin (DI). T1DM was induced by streptozotocin; insulin was administered daily (4IU). The animals were euthanized 35 days after induction. The tibiae were removed and analyzed using macroscopic, micro-computed tomography (micro-CT) and three-point bending. The macroscopic analysis measured proximal-distal length (PD), antero-posterior thickness (AP) of proximal (AP-P) and distal (AP-D) epiphysis, and lateral-medial thickness (LM) of proximal (LM-P) and distal (LM-D) epiphysis. Micro-CT analysis closed porosity, tissue mineral density, and cortical thickness. The three-point bending test measured maximum strength, energy, and stiffness. RESULTS: The macroscopic analysis showed that D presented smaller measures of length and thickness (AP and AP-P) than ND and DI. More extensive measurements were observed of LM and AP-D thickness in DI than in D. In micro-CT, DI showed larger cortical thickness than D. Mechanical analysis showed lower strength in D than in other groups. CONCLUSIONS: T1DM reduces bone growth and mechanical strength. Insulin therapy in diabetic rats improved bone growth and fracture resistance, making diabetic bone similar to normoglycemic animals.

Platelet-Rich Fibrin Progressive Protocol: Third Generation of Blood Concentrates.

PURPOSE: Platelet-rich fibrin (PRF) has been used in several fields of dentistry to improve tissue healing. However, PRF from glass tubes results in a limited number of small membranes, increasing clinical difficulty and work time. The aim of this study was to evaluate cell and platelet amounts and biomechanical strength of PRF-giant membranes produced from plastic tubes without additives. MATERIAL AND METHODS: The investigators designed an ex vivo study, to compare 3 different centrifugation protocols for obtaining PRF: 700 × g/12 minutes (leukocyte and PRF [L-PRF]), 350 × g/14 minutes (GM350), and 60-700 × g more than 15 minutes total (progressive PRF [PRO-PRF]). We collected blood samples from 5 volunteers aged 25-54 years, over 3 different time periods (triplicate and paired study). From each venipuncture, 4 mL of blood was collected in vacutainers with ethylenediamine tetraacetic acid (EDTA) and approximately 104 mL in 12 plastic tubes without additives, which were separated into 3 groups, as per the centrifugation protocols (n = 5): L-PRF, GM350, and PRO-PRF. The PRF from the tubes of the same protocol was aspirated and 9 mL were placed in polylactic acid (PLA) forms and 3 mL were placed in a glass receptacle. The membranes from PLA forms were tested for tensile strength and the membranes from glass receptacles were evaluated by histomorphometry, while platelets and leukocytes were counted for those in tubes with EDTA. Statistical analyses were performed using Shapiro-Wilk normality test and then a one-way repeated measures analysis followed by Tukey multiple comparisons test (α < 0.05). RESULTS: In tensile analyses, PRO-PRF (0.85 ± 0.23 N) showed a significantly higher maximum breaking strength than L-PRF (0.61 ± 0.26 N, P = .01) and GM350 (0.58 ± 0.23 N, P < .01). The histomorphometry revealed no significant statistical difference in cell counts between the groups (P = .52). Furthermore, there was no significant difference between the leukocyte (P = .25) and platelet counts (P = .59) in whole blood between the groups. CONCLUSION: The progressive protocol (PRO-PRF) enabled the production of PRF giant membranes with greater tensile strength and adequate cell distribution. Moreover, it allows biomaterial incorporation during production and enables clinical control of membrane thickness and size as per the surgical procedure.

Automated bone healing evaluation: New approach to histomorphometric analysis.

This study aimed to assess different approaches for bone healing evaluation on histological images and to introduce a new automatic evaluation method based on segmentation with distinct thresholds. We evaluated the hyperbaric oxygen therapy (HBO) effects on bone repair in type 1 diabetes mellitus rats. Twelve animals were divided into four groups (n = 3): non-diabetic, non-diabetic + HBO, diabetic, and diabetic + HBO. Diabetes was induced by intravenous administration of streptozotocin (50 mg/kg). Bone defects were created in femurs and HBO was immediately started at one session/day. After 7 days, the animals were euthanized, femurs were removed, demineralized, and embedded in paraffin. Histological sections were stained with hematoxylin and eosin (HE) and Mallory's trichrome (MT), and evaluated using three approaches: (1) conventional histomorphometric analysis (HE images) using a 144-point grid to quantify the bone matrix; (2) a semi-automatic method based on bone matrix segmentation to assess the bone matrix percentage (MT images); and (3) automatic approach, with the creation of a plug-in for ImageJ software. The time required to perform the analysis in each method was measured and subjected to Bland-Altman statistical analysis. All three methods were satisfactory for measuring bone formation and were not statistically different. The automatic approach reduced the working time compared to visual grid and semi-automated method (p < .01). Although histological evaluation of bone healing was performed successfully using all three methods, the novel automatic approach significantly shortened the time required for analysis and had high accuracy.

Type I Diabetes Mellitus and Insulin Therapy on Bone Microarchitecture, Composition and Mechanical Properties.

BACKGROUND: The aim of this study was to evaluate the microarchitecture, composition and mechanical properties of cortical bone of rats with type I diabetes mellitus (TIDM) and submitted to insulin therapy (IT). METHODS: Thirty rats were divided into three groups (n=10): non-diabetic, diabetic and diabetic+insulin. TIDM was induced by intravenous injection of streptozotocin. In diabetic+insulin group, 4IU insulin was administered twice per day (1I U at 7 am and 3I U at 7 pm). The animals were euthanized five weeks after TIDM induction; the tibiae were removed and submitted to microcomputed tomography (micro-CT, 8 µm), fourier transform infrared spectroscopy (FTIR) and dynamic microhardness indentation. RESULTS: Micro-CT analysis showed that diabetic group had lower bone surface/tissue volume ratio (BS/BV) (p=0.018), cortical thickness (Ct.Th) (p<0.001) and degree of anisotropy (Ct.DA) (p=0.034) values compared to non-diabetic group. The diabetic group showed lower Ct.Th than diabetic + insulin group (p=0.018). The non-diabetic group had lower fractal dimension (Ct.FD) values compared to diabetic groups (p<0.001). The ATR-FTIR analyses showed lower values for all measured parameters in the diabetic group than the non-diabetic group (amide I ratio: p=0.046; crystallinity index: p=0.038; matrix:mineral ratios - M:MI: p=0.006; M:MIII: p=0.028). The diabetic+ insulin group showed a lower crystallinity index (p=0.022) and M:MI ratio (p=0.002) than nondiabetic and diabetic groups, respectively. The diabetic group showed lower Vickers hardness values than non-diabetic (p<0.001) and diabetic+insulin (p=0.003) groups. CONCLUSION: TIDM negatively affects bone microarchitecture, collagen maturation, mineralization and bone microhardness. Moreover, insulin minimized the effect of TIDM on cortical thickness and organic/mineral matrix.

Implant site preparation-related bone microdamage: An alternative ex vivo analysis of drilling speed protocols / Microdanos ósseos gerados na preparação do sítio de instalação de implantes: uma alternativa para análise ex vivo de diferentes protocolos de velocidade de perfuração

ABSTRACT Objective: The aim of this study was to evaluate bone microdamage in sites prepared for implant placement by using an ex vivo model with three drilling rotation speeds. Methods: Bovine bone ribs were used for the creation of 18 osteotomy sites at different rotation speeds: 1200 rpm, 800 rpm, and 400 rpm. Specimens were stained with xylenol orange and prepared for histological analysis by using fluorescence and polarized light microscopies. Bone microdamage was evaluated by number and based on total bone area, as follows: microfracture density (Fr.D = n/mm2), microcrack morphology (diffuse or linear), and density (Cr.D = n/mm2), and presence of bone chips. To complement the analysis, linear microcracks were assessed by using confocal microscopy for three-dimensional visualization. Results: Bone microdamage on the osteotomy sites included microcracks, diffuse damages, microfracture, and bone chip formation. There was an association between bone microdamage and cancellous bone (p 0.0016), as well as a positive correlation between Fr.D and Cr.D (p 0.05, r 0.54). BM occurrence was not different between the three rotation speeds. In 3D, the height of the microcrack depth was 60.81 µm. Conclusion: Bone microdamage occurs during osteotomy, and the ex vivo model used was effective for the assessment of these biomechanical parameters. In addition, microdamage was not influenced by the drilling rotation speed in this experimental condition.

RESUMO Objetivo: O objetivo desse estudo foi avaliar os microdanos ósseos em locais preparados para a instalação de implantes utilizando um modelo ex vivo acessando três velocidades de rotação de perfuração. Métodos: Fragmentos ósseos de costela bovina foram utilizados para a criação de 18 sítios de osteotomia em diferentes velocidades de rotação: 1200 rpm, 800 rpm e 400 rpm. As amostras foram coradas com Alaranjado de Xilenol e preparadas para análise microscópica em fluorescência e luz polarizada. Os microdanos ósseos foram avaliados em número e calibrados com base na área total óssea: densidade de microfraturas (Fr.D = n/mm2), morfologia (difusa ou linear) e densidade de microtrincas (Cr.D = n / mm2) e presença de espículas ósseas. Para complementar a análise, microtrincas lineares foram avaliadas por meio de microscopia confocal para visualização tridimensional. Resultados: Os microdanos ósseos incluíram microtrincas, danos difusos, microfraturas e formação de espículas. Houve uma associação entre MO e localização em osso esponjoso (p=0,0016), bem como uma correlação positiva entre Fr.D e Cr.D (p=0,05, r = 0,54). A ocorrência de microdanos ósseos não foi diferente entre as três velocidades de rotação utilizadas. Em 3D, a profundidade maior da microfissura atingiu 60,81 µm. Conclusão: Microdanos ósseos ocorrem durante a osteotomia e podem ser acessados em um modelo ex vivo na condução de experimentos em biomecânica. Sugere-se que a presença de microdanos não é influenciada pela velocidade de rotação durante a perfuração.

Automatic Segmentation of Bone Canals in Histological Images.

The literature provides many works that focused on cell nuclei segmentation in histological images. However, automatic segmentation of bone canals is still a less explored field. In this sense, this paper presents a method for automatic segmentation approach to assist specialists in the analysis of the bone vascular network. We evaluated the method on an image set through sensitivity, specificity and accuracy metrics and the Dice coefficient. We compared the results with other automatic segmentation methods (neighborhood valley emphasis (NVE), valley emphasis (VE) and Otsu). Results show that our approach is proved to be more efficient than comparable methods and a feasible alternative to analyze the bone vascular network.

Effect of ionizing radiation and cariogenic biofilm challenge on root-dentin caries.

OBJECTIVES: To evaluate the effect of ionizing radiation and cariogenic biofilm challenge using two continuous flow models, normal and reduced salivary flow, on the development of initial root-dentin caries lesions. MATERIALS AND METHODS: Microcosm biofilms were grown under two salivary flow rates (0.06 and 0.03 mL min-1) and exposed to 5% sucrose (3 × daily, 0.25 mL min-1, 6 min) dripped over non-irradiated and irradiated root-dentin blocks for up to 7 days. The vibration modes of root dentin, matrix/mineral (M/M), and carbonate/mineral (C/M) ratios were evaluated by FTIR. The mineral density was assessed by micro-CT. RESULTS: With normal salivary flow, FTIR revealed an increase in the organic matrix (amide III) and a decrease in the mineral phase (ν4, ν2 PO43-, AII + ν2 CO32-, C/M) in caries lesions. Irradiated dentin exhibited a reduction in the mineral phase (ν1, ν3 PO43-, ν2 CO32-, C/M). Differences in mineral densities were not significant. With reduced salivary flow, FTIR also revealed increased organic matrix (amide III) for irradiated caries lesions and decrease in mineral phase (v4, v2 PO43-, v2 CO32-, and C/M) in caries lesions. ν1, ν3 PO43- precipitated on the surface of irradiated dentin and a lower mineral density was observed. CONCLUSIONS: Initial caries lesions differed between non-irradiated and irradiated dentin and between normal and reduced salivary flow rates. Significant mineral loss with exposure of the organic matrix and low mineral density were observed for irradiated dentin with a reduced salivary flow rate. CLINICAL RELEVANCE: Ionizing radiation associated with a reduced salivary flow rate enhanced the progression of root-dentin caries.

Biomechanical and morphological changes produced by ionizing radiation on bone tissue surrounding dental implant.

OBJECTIVE: This study analyzed the effect of ionizing radiation on bone microarchitecture and biomechanical properties in the bone tissue surrounding a dental implant. METHODOLOGY: Twenty rabbits received three dental morse taper junction implants: one in the left tibia and two in the right tibia. The animals were randomized into two groups: the nonirradiated group (control group) and the irradiated group, which received 30 Gy in a single dose 2 weeks after the implant procedure. Four weeks after the implant procedure, the animals were sacrificed, and the implant/bone specimens were used for each experiment. The specimens (n=10) of the right tibia were examined by microcomputed tomography to measure the cortical volume (CtV, mm3), cortical thickness (CtTh, mm) and porosity (CtPo, %). The other specimens (n=10) were examined by dynamic indentation to measure the elastic modulus (E, GPa) and Vickers hardness (VHN, N/mm2) in the bone. The specimens of the left tibia (n=10) were subjected to pull-out tests to calculate the failure load (N), displacement (mm) up to the failure point and interface stiffness (N/mm). In the irradiated group, two measurements were performed: close, at 1 mm surrounding the implant surface, and distant, at 2.5 mm from the external limit of the first measurement. Data were analyzed using one-way ANOVA, Tukey's test and Student's t-test (α=0.05). RESULTS: The irradiated bone closer to the implant surface had lower elastic modulus (E), Vickers hardness (VHN), Ct.Th, and Ct.V values and a higher Ct.Po value than the bone distant to the implant (P<0.04). The irradiated bone that was distant from the implant surface had lower E, VHN, and Ct.Th values and a higher Ct.Po value than the nonirradiated bone (P<0.04). The nonirradiated bone had higher failure loads, displacements and stiffness values than the irradiated bone (P<0.02). CONCLUSION: Ionizing radiation in dental implants resulted in negative effects on the microarchitecture and biomechanical properties of bone tissue, mainly near the surface of the implant.

Impact of Childhood Obesity on Bone Metabolism.

Childhood obesity is a considerable worldwide health problem and a major risk factor for several chronic diseases. Fat rich diets result in altered serum levels of lipids, cytokines and hormonal factors, which influence skeletal acquisition and promote microstructural and mechanical behavior changes in bone, especially to bone quality and quantity. However, the possible longterm implications of high-fat diets in childhood are controversial. Despite not fully understood, multiple signaling pathways which support bone tissue homeostasis are altered under hyperlipidic conditions, including RANKL/RANK/OPG, PPAR-γ/Alox5/5-LO, leptin/IGF-I/AGE, ApoE/Lrp-1, Thy-1, IL-6, TNFα, calcium, vitamin D and K metabolism. Moreover, the expression of reactive oxygen species is also modified. Considering the importance of this subject, the aim of this review was to explore the mechanisms of bone formation affected by obesity during childhood during childhood.

The effect of hyperbaric oxygen therapy on bone macroscopy, composition and biomechanical properties after ionizing radiation injury.

BACKGROUND: Radiotherapy used in tumor treatment compromises vascularization of bone tissue. Hyperbaric oxygenation (HBO) increases oxygen availability and improves vascularization, minimizing the deleterious effects of ionizing radiation (IR). Therefore, the aim of this study was to evaluate HBO therapy effect on bone macroscopy, composition and biomechanical properties after IR damage. METHODS: Twenty male Wistar rats weighing 300 ± 20 g (10 weeks of age) were submitted to IR (30 Gy) to the left leg, where the right leg was not irradiated. After 30 days, ten animals were submitted to HBO therapy, which was performed daily for 1 week at 250 kPa for 90-min sessions. All animals were euthanized 37 days after irradiation and the tibia were separated into four groups (n = 10): from animals without HBO - right tibia Non-irradiated (noIRnoHBO) and left tibia Irradiated (IRnoHBO); and from animals with HBO - right tibiae Non-irradiated (noIRHBO) and left tibia Irradiated (IRHBO). The length (proximal-distal) and thickness (anteroposterior and mediolateral) of the tibiae were measured. Biomechanical analysis evaluated flexural strength and stiffness. Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy (ATR-FTIR) was used to calculate the amide I ratio, crystallinity index, and matrix to mineral ratios. RESULTS: In the macroscopic and ATR-FTIR analysis, the IRnoHBO showed lower values of length, thickness and amide I ratio, crystallinity index and matrix to mineral ratios compared to noIRnoHBO (p < 0.03). IRnoHBO showed no statistical difference compared to IRHBO for these analyses (p > 0.05). Biomechanics analysis showed that the IRnoHBO group had lower values of flexural strength and stiffness compared to noIRnoHBO and IRHBO groups (p < 0.04). In addition, the noIRHBO group showed higher value of flexural strength when compared to noIRnoHBO and IRHBO groups (p < 0.02). CONCLUSIONS: The present study concluded that IR arrests bone development, decreases the collagen maturation and mineral deposition process, thus reducing the flexural strength and stiffness bone mechanical parameters. Moreover, HBO therapy minimizes deleterious effects of irradiation on flexural strength and the bone stiffness analysis.

Ionizing radiation and bone quality: time-dependent effects.

BACKGROUND: The aim of this study was to evaluate the ionizing radiation (IR) effects on rat bone 30 and 60 days after irradiation. METHODS: Wistar rats were submitted to IR (30 Gy) on the left leg and were euthanized after 30 and 60 days. The legs were divided into four groups according to the treatment and euthanization time: C30 and C60 (right leg-without IR), IR30 and IR60 (left leg-with IR). RESULTS: CT analysis showed more radiodensity in C60 compared with other groups, and IR60 showed more radiodensity than IR30. In histomorphometric analysis, C30 showed lower bone matrix values compared with IR30 and C60. Lacunarity analyses showed more homogeneous bone channel distribution in C30 than IR30. ATR-FTIR showed decrease in ratio of mature and immature crosslinks in IR30 compared with C30. Crystallinity Index was decrease in IR60 compared with C60. The Amide III + Collagen/HA ratio was increased in C60 compared with C30; however this ratio decreased in IR60 compared with IR30. Biomechanical analysis showed lower values in IR groups in both time. CONCLUSIONS: IR damaged bone quality and decreased stiffness. Moreover, the results suggested that the deleterious effects of IR increased in the late time points.

Effect of ionizing radiation after-therapy interval on bone: histomorphometric and biomechanical characteristics.

OBJECTIVES: This study aimed to evaluate the effects of radiotherapy on biomechanical, histomorphometric, and microstructural characteristics of bone, in diverse periods, compared with intact bone tissue. MATERIALS AND METHODS: Eighteen adult male New Zealand rabbits were treated with a single radiation dose of 30 Gy. The animals were randomly divided into six groups: NoIr, control group, no radiation, and five irradiated groups sacrificed after 24 h (Ir24h), 7 (Ir7d), 14 (Ir14d), 21 (Ir21d), and 28 (Ir28d) days. After these periods, the animals were sacrificed and their tibias (n = 6) evaluated using three-point bending test to calculate the ultimate force, work to failure, and bone stiffness. Dynamic indentation test was used to quantify Vickers hardness and elasticity modulus of bone tissue. Micro-CT was used to analyze the cortical volume (CtV), cortical thickness (CtTh), and porosity (Ct.Po). Histomorphometric assessment was based on the lacunarity of bone tissue. Data were analyzed using one-way ANOVA and Kruskal-Wallis tests followed by Tukey, Dunnet, and Dunn's post-tests (P < 0.05). RESULTS: The ultimate force, work to failure, stiffness, elastic modulus, and Vickers hardness values of irradiated bone were significantly lower that non-irradiated bone. Irradiated bone showed significantly lower CtTh and CtV values and higher CtPo than non-irradiated bone. No significant difference was found for lacunarity between non-irradiated bone and irradiated bone. CONCLUSIONS: Ionizing radiation decreases normal anisotropy on microarchitecture of cortical bone, and increases bone fragility compared with non-irradiated bone. Further, these changes were seen after longer periods (e.g., 14 and 21 days), and not immediately after radiation therapy. CLINICAL RELEVANCE: The radiotherapy reduces bone mechanical properties and the normal structure of organic and inorganic bone matrix. For studying the protocols to protect the radiotherapy effect using rabbit model, the use of the sacrificing period between 14 and 21 days is recommended.

Influence of hyperbaric oxygen on biomechanics and structural bone matrix in type 1 diabetes mellitus rats.

BACKGROUND: The aim of this study was to evaluate the biomechanics and structural bone matrix in diabetic rats subjected to hyperbaric oxygen therapy (HBO). METHODS: Twenty-four male rats were divided into the following groups: Control; Control + HBO; Diabetic, and Diabetic + HBO. Diabetes was induced with streptozotocin (STZ) in the diabetic Groups. After 30 days, HBO was performed every 48h in HBO groups and all animals were euthanized 60 days after diabetic induction. The femur was submitted to a biomechanical (maximum strength, energy-to-failure and stiffness) and Attenuated Total Reflectance Fourier transform infrared (ATR-FTIR) analyses (crosslink ratio, crystallinity index, matrix-to-mineral ratio: Amide I + II/Hydroxyapatite (M:MI) and Amide III + Collagen/HA (M:MIII)). RESULTS: In biomechanical analysis, diabetic animals showed lower values of maximum strength, energy and stiffness than non-diabetic animals. However, structural strength and stiffness were increased in groups with HBO compared with non-HBO. ATR-FTIR analysis showed decreased collagen maturity in the ratio of crosslink peaks in diabetic compared with the other groups. The bone from the diabetic groups showed decreased crystallinity compared with non-diabetic groups. M:MI showed no statistical difference between groups. However, M:MIII showed an increased matrix mineral ratio in diabetic+HBO and control+HBO compared with control and diabetic groups. Correlations between mechanical and ATR-FTIR analyses showed significant positive correlation between collagen maturity and stiffness. CONCLUSIONS: Diabetes decreased collagen maturation and the mineral deposition process, thus reducing biomechanical properties. Moreover, the study showed that HBO improved crosslink maturation and increased maximum strength and stiffness in the femur of T1DM animals.

Hyperbaric Oxygen therapy effects on bone regeneration in Type 1 diabetes mellitus in rats.

PURPOSE: The aim of this study was evaluate the effect of HBO on diabetic rats. MATERIALS AND METHODS: Twenty rats were distributed into four groups (n = 5): Control (C); Control + HBO (CH); Diabetes (D) and Diabetes + HBO (DH). Diabetes was induced by streptozotocin, and bone defects were created in both femurs in all animals. HBO therapy began immediately after surgery and was performed daily in the CH and DH groups. After 7 days, the animals were euthanized. The femurs were removed, demineralized, embedded in paraffin, and histologic images were analyzed. RESULTS: Qualitative histologic analyses showed more advanced stage bone regeneration in control groups (C and CH) compared with diabetic groups (D and DH). Histomorphometric analysis showed significantly increased bone neoformation in CH compared with the other groups (p < 0.001). Diabetic Group (D) showed decreased bone neoformation compared with non-diabetic groups (C and CH) (p < 0.001); however DH did not differ from C Group (p > 0.05). The mast cell population increased in CH compared with the other groups (C, D, and DH) (p < 0.05). The mast cell population did not differ between D and DH Groups. CONCLUSIONS: This study showed that HBO therapy improved early bone regeneration in diabetic rats and increased the mast cell population only in non-diabetic animals. HBO was shown to be important treatment for minimizing deleterious effects of diabetes on bone regeneration.

High doses of ionizing radiation on bone repair: is there effect outside the irradiated site?

Local ionizing radiation causes damage to bone metabolism, it reduces blood supply and cellularity over time. Recent studies indicate that radiation promotes biological response outside the treatment field. The aim of this study was to investigate the effects of ionizing radiation on bone repair outside the irradiated field. Ten healthy male Wistar rats were used; and five animals were submitted to radiotherapy on the left femur. After 4 weeks, in all animals were created bone defects in the right and left femurs. Seven days after surgery, animals were euthanized. The femurs were removed and randomly divided into 3 groups (n=5): Control (C) (right femur of the non-irradiated animals); Local ionizing radiation (IR) (left femur of the irradiated animals); Contralateral ionizing radiation (CIR) (right femur of the irradiated animals). The femurs were processed and embedded in paraffin; and bone histologic sections were evaluated to quantify the bone neoformation. Histomorphometric analysis showed that there was no significant difference between groups C (24.6±7.04) and CIR (25.3±4.31); and IR group not showed bone neoformation. The results suggest that ionizing radiation affects bone repair, but does not interfere in bone repair distant from the primary irradiated site.

The effects of ionizing radiation on the growth plate in rat tibiae.

The deleterious effects of ionizing radiation on the growth plate continue to be cause for concern. This study evaluated the ionizing radiation effects on bone development and growth plate in the tibia of rats. All animals were submitted to ionizing radiation on the left leg. The animals were divided into two groups and euthanized 30 and 60 days after radiation. The tibiae were removed and separated into groups: control 30 days, irradiated 30 days, control 60 days and irradiated 60 days. Animals in each group (n = 7) were used for macroscopic and histological analysis. The irradiated tibiae showed arrested growth, angular deformity and limb length discrepancy when compared with nonirradiated tibiae. There was statistical difference between control and radiation groups in all the parameters analyzed, except in the lateral-medial thickness of the distal epiphysis. Histological analysis showed evident changes in the growth plate, which was thicker in the Groups irradiated for 30 days, and irradiated for 60 days, compared with their respective controls. The growth plate showed wide areas with disorganized zones of chondrocytes and severely reduced calcification zone. It was concluded that ionizing radiation damaged the growth plate, compromised the endochondral ossification process, and resulted in complete arrest of bone development.

The Effect of Local Delivery Doxycycline and Alendronate on Bone Repair.

The aim of the present study was to investigate the local effect of 10% doxycycline and 1% alendronate combined with poly(lactic-co-glycolic acid) (PLGA) on bone repair. Thirty rats were divided into three groups, as follows: control group (CG), drug group (DG), and vehicle-PLGA group (VG). Bone defect was created in the right femur and filled with the following: blood clot (CG); PLGA gel, 10% doxycycline and 1% alendronate (DG); or vehicle-PLGA (VG). The animals were euthanized 7 or 15 days after surgery. Bone density, bone matrix and number of osteoclasts were quantified. At 7 days, the findings showed increased density in DG (177.75 ± 76.5) compared with CG (80.37 ± 27.4), but no difference compared with VG (147.1 ± 41.5); no statistical difference in bone neoformation CG (25.6 ± 4.8), VG (27.8 ± 4), and DG (18.9 ± 7.8); and decrease osteoclasts in DG (4.6 ± 1.9) compared with CG (26.7 ± 7.4) and VG (17.3 ± 2.7). At 15 days, DG (405.1 ± 63.1) presented higher density than CG (213.2 ± 60.9) and VG (283.4 ± 85.8); there was a significant increase in percentage of bone neoformation in DG (31.5 ± 4.2) compared with CG (23 ± 4), but no difference compared with VG (25.1 ± 2.9). There was a decreased number of osteoclasts in DG (20.7 ± 4.7) and VG (29.5 ± 5.4) compared with CG (40 ± 9.4). The results suggest that the association of 10% doxycycline and 1% alendronate with PLGA-accelerated bone repair.