João Paulo Mardegan Issa

Sustained Release Carriers Used to Delivery Bone Morphogenetic Proteins in the Bone Healing Process

Bone morphogenetic proteins (BMPs) are multi‐functional growth factors belonging to the transforming growth factor β superfamily, especially BMP‐2, induce bone formation in vivo, and clinical application in repair of bone fractures and defects is expected. However, appropriate systems to delivery BMPs for practical use need to be developed with the objective to heal cartilage and bone‐related diseases in medical, dental and veterinary practice. Thus, the aim of this article was to present an overview of the principals carriers used to delivery BMPs and alternative delivery systems for these proteins.

Ultrastructural analysis of the low level laser therapy effects on the lesioned anterior tibial muscle in the Gerbil

Low level laser therapy (LLLT) is known for its positive results but studies on the biological and biomodulator characteristics of the effects produced in the skeletal muscle are still lacking. In this study the effects of two laser dosages, 5 or 10 J/cm(2), on the lesioned tibial muscle were compared. Gerbils previously lesioned by 100 g load impact were divided into three groups: GI (n=5) controls, lesion non-irradiated; GII (n=5), lesion irradiated with 5 J/cm(2) and GIII (n=5), lesion irradiated with 10 J/cm(2), and treated for 7 consecutive days with a laser He-Ne (lambda=633 nm). After intracardiac perfusion, the muscles were dissected and reduced to small fragments, post-fixed in 1% osmium tetroxide, dehydrated in increasing alcohol concentrations, treated with propylene oxide and embedded in Spurr resin at 60 degrees C. Ultrafine cuts examined on a transmission electron microscope (Jeol 1010) revealed in the control GI group a large number of altered muscle fibers with degenerating mitochondria, intercellular substance containing degenerating cell fragments and budding blood capillaries with underdeveloped endothelial cells. However, groups GII and GIII showed muscle fibers with few altered myofibrils, regularly contoured mitochondria, ample intermembrane spaces and dilated mitochondrial crests. The clean intercellular substance showed numerous collagen fibers and capillaries with multiple abluminal processes, intraluminal protrusions and several pinocytic vesicles in endothelial cells. It was concluded that laser dosages of 5 or 10 J/cm(2) delivered by laser He-Ne (lambda=633 nm) during 7 consecutive days increase mitochondrial activity in muscular fibers, activate fibroblasts and macrophages and stimulate angiogenesis, thus suggesting effectivity of laser therapy under these experimental conditions.

Effects of low-level laser irradiation in ultrastructural morphology, and immunoexpression of VEGF and VEGFR-2 of rat masseter muscle

The present study evaluates by ultrastructural and immunohistochemical methods, the possible changes on muscular tissue affected by LLLI during a treatment, for example, in cases of temporomandibular joint disorders. Sixty male Wistar rats divided into 6 groups (n=10) received ten laser irradiations, with different energy densities (groups I-0; II-0.5; III-1.0; IV-2.5; V-5.0; and VI-20 J/cm(2)). Muscles were removed and processed for transmission electron microscopic and immunohistochemical (VEGF and VEGFR-2) analyses. Captured photomicrographs of immunohistochemistry and transmission electron microscopy were evaluated. It was observed in the irradiated muscles, mitochondria of different shapes and sizes, with increased plasticity evidenced by organelles in fusion, division and the presence of elongated structures with characteristics of mitochondria, proximity with the dilated sarcoplasmatic reticulum, suggesting organelles with large amounts of energy, and the presence of cytoplasmic protrusions in the capillaries with high dosages. All studied groups showed immunostainings for both markers (VEGF and VEGFR-2), but in general those who received higher doses also showed the markings more pronounced, suggesting dose-dependent biomodulation. It was concluded that the LLLI was able to modify the ultrastructural characteristics and immunohistochemical pattern of VEGF and VEGFR-2 in the masseter muscle of rats.

Use of checkerboard DNA-DNA hybridization to evaluate the internal contamination of dental implants and comparison of bacterial leakage with cast or pre-machined abutments.

AIMS To evaluate the checkerboard DNA-DNA hybridization method for detection and quantitation of bacteria from the internal parts of dental implants and to compare bacterial leakage from implants connected either to cast or to pre-machined abutments. MATERIALS AND METHODS Nine plastic abutments cast in a Ni-Cr alloy and nine pre-machined Co-Cr alloy abutments with plastic sleeves cast in Ni-Cr were connected to Branemark-compatible implants. A group of nine implants was used as control. The implants were inoculated with 3 microl of a solution containing 10(8) cells/ml of Streptococcus sobrinus. Bacterial samples were immediately collected from the control implants while assemblies were completely immersed in 5 ml of sterile Tripty Soy Broth (TSB) medium. After 14 days of anaerobic incubation, occurrence of leakage at the implant-abutment interface was evaluated by assessing contamination of the TSB medium. Internal contamination of the implants was evaluated with the checkerboard DNA-DNA hybridization method. RESULTS DNA-DNA hybridization was sensitive enough to detect and quantify the microorganism from the internal parts of the implants. No differences in leakage and in internal contamination were found between cast and pre-machined abutments. Bacterial scores in the control group were significantly higher than in the other groups (P<0.05). CONCLUSION Bacterial leakage through the implant-abutment interface does not significantly differ when cast or pre-machined abutments are used. The checkerboard DNA-DNA hybridization technique is suitable for the evaluation of the internal contamination of dental implants although further studies are necessary to validate the use of computational methods for the improvement of the test accuracy.

Evaluation of rhBMP-2 and Natural Latex as Potential Osteogenic Proteins in Critical Size Defects by Histomorphometric Methods

This in vivo study evaluated the osteogenic potential of two proteins, recombinant human bone morphogenetic protein‐2 (rhBMP‐2) and a protein extracted from natural latex (Hevea brasiliensis, P‐1), and compared their effects on bone defects when combined with a carrier or a collagen gelatin. Eighty‐four (84) Wistar rats were divided into two groups, with and without the use of collagen gelatin, and each of these were divided into six treatment groups of seven animals each. The treatment groups were: (1) 5 μg of pure rhBMP‐2; (2) 5 μg of rhBMP‐2/monoolein gel; (3) pure monoolein gel; (4) 5 μg of pure P‐1; (5) 5 μg of P‐1/monoolein gel; (6) critical bone defect control. The animals were anesthetized and a 6 mm diameter critical bone defect was made in the left posterior region of the parietal bone. Animals were submitted to intracardiac perfusion after 4 weeks and the calvaria tissue was removed for histomorphometric analysis. In this experimental study, it was concluded that rhBMP‐2 allowed greater new bone formation than P‐1 protein and this process was more effective when the bone defect was covered with collagen gelatin (P < 0.05). Anat Rec, 2010.

Bacterial Culture and DNA Checkerboard for the Detection of Internal Contamination in Dental Implants

PURPOSE The aim of this in vitro study was to evaluate the bacterial leakage along the implant-abutment interface by the conventional bacterial culture and DNA Checkerboard hybridization method. MATERIALS AND METHODS Twenty Branemark-compatible implants with a 3.75-mm diameter and external hexagonal platform were randomly placed in two groups of ten implant-abutment assemblies each. One group was used to analyze bacterial counts by DNA Checkerboard hybridization and the other by a conventional bacterial culture. Suspensions of Fusobacterium nucleatum (3 microl) were injected into the grooved internal cylinders of each implant assembly, and the abutment was connected by a 32 Ncm torque. The combined implant-abutments were individually placed in tubes containing the CaSaB culture medium and incubated in a bacteriological constant temperature oven for 14 days. The samples were observed daily as to the presence of turbidity, and after the designated time the microorganisms were collected from the implant interiors and analyzed by the two methods. RESULTS After 14 days, six implant-abutment assemblies showed turbidity. Both methods indicated reduced microorganism counts in samples from the interior of the implant-abutment assemblies after incubation in the culture medium; however, the number of counts of F. nucleatum was higher by the DNA Checkerboard method when compared to the group analyzed by conventional bacterial cultures (p < 0.05). CONCLUSION The DNA Checkerboard method was shown to be more sensitive than conventional cultures in the detection of microorganisms.

Matrix metalloproteinases are involved in cardiovascular diseases.

This MiniReview describes the essential biochemical and molecular aspects of matrix metalloproteinases (MMPs) and briefly discusses how they engage in different diseases, with particular emphasis on cardiovascular diseases. There is compelling scientific evidence that many MMPs, especially MMP-2, play important roles in the development of cardiovascular diseases; inhibition of these enzymes is beneficial to many cardiovascular conditions, sometimes precluding or postponing end-organ damage and fatal outcomes. Conducting comprehensive discussions and further studies on how MMPs participate in cardiovascular diseases is important, because inhibition of these enzymes may be an alternative or an adjuvant for current cardiovascular disease therapy.

Effect of recombinant human bone morphogenetic protein‐2 on bone formation in the acute distraction osteogenesis of rat mandibles

BACKGROUND Distraction osteogenesis (DO) is a method of producing new bone directly from the osteotomy site by gradual traction of the divided bone fragments. AIM The purpose of the present study was to evaluate histomorphometrically whether acute DO would constitute a viable alternative to the conventional continuous distraction treatment and also to verify the capacity of a recombinant human BMP (rhBMP-2) associated with monoolein gel to stimulate bone formation in the acute distraction process. MATERIALS AND METHODS Forty-eight Wistar rats were assigned to three groups: Group 1, treated at a conventional continuous distraction rate (0.5 mm/day), Group 2, treated with acute distraction of 2.5 mm at the time of the surgical procedure, and Group 3, subjected to acute distraction associated with rhBMP-2. The animals from each experimental group were killed at the end of the second or fourth post-operative weeks and the volume fraction of newly formed bone trabeculae was estimated in histological images by a differential point-counting method. RESULTS The results showed that after 2 and 4 weeks, bone volumes in the rhBMP-2 group were significantly higher than in the other groups (P<0.05), but no significant difference was observed in the volume fraction of newly formed bone between the continuous and acute DO groups. CONCLUSION In conclusion, the study indicates that rhBMP-2 can enhance the bone formation at acute DO, which may potentially reduce the treatment period and complications related to the distraction procedure.

Biomaterials Applied to the Bone Healing Process

Una innovacion reciente en medicina es la utilizacion de biomatenales en la reparacion de defectos oseos. Este trabajo tiene como objetivo presentar una actualizacion de los usos y aplicaciones de biomateriales y de sus diversos constituyentes mas empleados en la reparacion de los huesos

Morphodigital Study of the Mandibular Trabecular Bone in Panoramic Radiographs

El objetivo de este estudio fue correlacionar las caracteristicas morfodigitales del trabeculado oseo mandibular para de esta manera, verificar indicios de senales de osteoporosis, utilizando para esto, radiografias panoramicas de pacientes de ambos sexos. Los resultados del estudio mostraron diferencias entre los grupos masculino y femenino, en la cuantificacion de hueso trabecular mandibular (p 0,05). El grupo masculino presento un padron oseo trabecular mas compleo y fuerte que el grupo femenino. Este hallazgo puede ser un importante indicador en cuanto a los riesgos de osteoporosis, por ejemplo, siendo posible afirmar que los huesos de los individuos del grupo masculino estan menos sujetos a riesgo de fractura por osteoporosis