Mamie Mizusaki Iyomasa

Quercetin in w/o microemulsion: In vitro and in vivo skin penetration and efficacy against UVB-induced skin damages evaluated in vivo

The present study evaluated the potential of a w/o microemulsion as a topical carrier system for delivery of the antioxidant quercetin. Topical and transdermal delivery of quercetin were evaluated in vitro using porcine ear skin mounted on a Franz diffusion cell and in vivo on hairless-skin mice. Skin irritation by topical application of the microemulsion containing quercetin, and the protective effect of the formulation on UVB-induced decrease of endogenous reduced glutathione levels and increase of cutaneous proteinase secretion/activity were also investigated. The w/o microemulsion increased the penetration of quercetin into the stratum corneum and epidermis plus dermis at 3, 6, 9 and 12h post-application in vitro and in vivo at 6h post-application. No transdermal delivery of quercetin occurred. By evaluating established endpoints of skin irritation (erythema formation, epidermis thickening and infiltration of inflammatory cells), the study demonstrated that the daily application of the w/o microemulsion for up to 2 days did not cause skin irritation. W/o microemulsion containing quercetin significantly prevented the UVB irradiation-induced GSH depletion and secretion/activity of metalloproteinases.

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.

Evaluation of Protective Effect of a Water-In-Oil Microemulsion Incorporating Quercetin Against UVB-Induced Damage in Hairless Mice Skin

PURPOSE In the present study, histological aspects were considered in order to evaluate the in vivo photoprotective effect of a w/o microemulsion containing quercetin against UVB irradiation-induced dermal damages. The toxicity in cell culture and the potential skin irritation resulting from topical application of this formulation were also investigated. METHODS Mouse dorsal surfaces were treated topically with 300 mg of the unloaded and quercetin-loaded (0.3%, w/w) microemulsions before and after exposure to UVB (2.87 J/cm2) irradiation. The untreated control groups irradiated and non-irradiated were also evaluated. UVB-induced histopathological changes as well as the photoprotective effect of this formulation were evaluated considering the parameters of infiltration of inflammatory cells, epidermis thickening (basale and spinosum layers) and collagen and elastic fiber contents. The cytotoxicity of the reported formulation was evaluated in L929 mice fibroblasts by MTT assay and the skin irritation was investigated after topical application of both unloaded and quercetin-loaded microemulsions once a day for 15 days. RESULTS The results demonstrated that the w/o microemulsion containing quercetin reduced the incidence of histological skin alterations, mainly the connective-tissue damage, induced by exposure to UVB irradiation, this allows the suggestion that protective effects of this formulation against UV-induced responses are not secondary to the interference of UV transmission (i.e., blocking the UVB radiation from being absorbed by the skin), as is usually done with UVB absorbers and sunscreens, but is instead due to different biological effects of this flavonoid. Furthermore, by evaluating the cytotoxic effect on L929 cells and histological aspects such as infiltration of inflammatory cells and epidermis thickness of hairless mice, the present study also demonstrated no toxicity of the proposed system. CONCLUSIONS Therefore, based on these mouse models, a detailed characterization of the w/o microemulsion incorporating quercetin effects as a photochemoprotective agent on human skin is thus indicated.

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.

Self-assembling gelling formulation based on a crystalline-phase liquid as a non-viral vector for siRNA delivery

Liquid crystalline systems (LCSs) form interesting drug delivery systems. These include in situ gelling delivery systems, which present several advantages for use as self-assembling systems for local drug delivery. The aim of this study was to develop and characterize in situ gelling delivery systems for local siRNA delivery. The influence of the components that form the systems was investigated, and the systems were characterized by polarized light microscopy, Small Angle X-ray Scattering (SAXS), swelling studies, assays of their ability to form a complex with genes and of the stability of the genes in the system, as well as assays of in situ gelling formation and local toxicity using an animal model. The system containing a mixture of monoglycerides (MO), oleylamine (OAM), propylene glycol (PG) and tris buffer (8.16:0.34:76.5:15, w/w/w/w) was considered the most appropriate for local siRNA delivery purposes. The molecular structure was characterized as hexagonal phase; the swelling studies followed a second order kinetic model and the water absorption was a fast process reaching equilibrium at 2 h. The system formed a complex with siRNA and remained in a stable form. The gel was formed in vivo after subcutaneous administration of a precursor fluid formulation in mice and was biodegradable in 30 days. The inflammatory process that took place was considered normal. Therefore, the developed liquid crystalline delivery system shows the appropriate characteristics for use as a local siRNA delivery method for gene 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.

Initial pulp changes during orthodontic movement: histomorphological evaluation

This study evaluated pulp changes in molars of rats submitted to tooth movement by application of a 0.4 N force. Twenty-five adult male Wistar rats (Rattus norvegicus, albinus) were randomly assigned to 5 groups (n=5), being one control group not submitted to force application, and four study groups of 6, 12, 24 and 72 h of force application. The study groups received a 5-mm long nickel-titanium closed coil spring, placed from the right maxillary first molar to the maxillary incisors of each animal. The coil spring was used for mesial inclination of the first molar. After the specific period of tooth movement of each study group, the animals were sacrificed and specimens containing the teeth submitted to movement were processed and stained with hematoxylin and eosin for histological analysis under light microscopy. The results demonstrated alteration of the odontoblastic layer, with hypertrophy of odontoblasts especially at the mesial area of the coronal pulp, edema of the pulp connective tissue in the central area of the pulp, and vascular alteration with accumulation of erythrocytes and leukocytes inside the vessels, especially at the mesial root of the moved teeth. These changes were less remarkable for the 72-h period. Thus, it may be concluded that tooth movement yielded pulpal tissue alterations compatible with an inflammatory process, which are reversible if the aggression is not more intense than the physiological limit of tissue tolerance.

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