Effects of Nutritional Deficiency of Boron on the Bones of the Appendicular Skeleton of Mice.

Scientific evidence has shown the nutritional importance of boron (B) in the remodeling and repair of cancellous bone tissue. However, the effects of the nutritional deficiency of B on the cortical bone tissue of the appendicular skeleton have not yet been described. Thus, a study was performed to histomorphometrically evaluate the density of osteocyte lacunae of cortical bone of mouse femora under conditions of nutritional deficiency of B and to analyze the effects of the deficiency on the biomechanical properties of mouse tibiae. Weaning, 21-day-old male Swiss mice were assigned to the following two groups: controls (B+; n = 10) and experimental (B-; n = 10). Control mice were fed a basal diet containing 3 mg B/kg, whereas experimental mice were fed a B-deficient diet containing 0.07 mg B/kg for 9 weeks. The histological and histomorphometric evaluations of the mice fed a B-deficient diet showed a decrease in the density of osteocyte lacunae in the femoral cortical bone tissue and the evaluation of biomechanical properties showed lower bone rigidity in the tibia.

Evaluation of the antibacterial effects of vancomycin hydrochloride released from agar-gelatin-bioactive glass composites.

The aim of this work was to evaluate the perfomance of agar-gelatin (AG) composites and AG-containing 45S5 bioactive glass (BG) microparticles (AGBG) in relation to their water uptake capacity, sustained release of a drug over time, and antibacterial effects. The composites were fabricated by the gel-casting method. To impart the local drug release capacity, vancomycin hydrochloride (VC) was loaded in the composites in concentrations of 0.5 and 1 mg ml(-1). VC release was assessed in distilled water at 37 °C up to 72 h and quantified spectrophotometrically. The antibacterial activity of composites was evaluated by the inhibition zone test and the plate count method. The experiments were performed in vitro up to 48 h on three staphylococcus strains: Staphylococcus aureus ATCC29213, S. aureus ATCC6538 and Staphylococcus epidermidis ATCC12228. The results showed that the addition of BG to AG composites did not affect the degree of water uptake. The release of VC was significantly affected by the presence of BG. VC release was higher from AGBGVC films than from AGVC ones over prolonged incubation times. Bacterial inhibition zones were found around the composites. The halos were larger when the cells were put in contact with AGVC composites than when they were put in contact with AGBGVC ones. Nevertheless, the viable count method demonstrated that the composites inhibited Staphylococcus cell growth with no statistical differences. In conclusion, the addition of BG did not reflect an improvement in the parameters studied. On the other hand, composites loaded with VC would have a role in prophylaxis against bacterial infection.

Effect of nano-sized bioactive glass particles on the angiogenic properties of collagen based composites.

Angiogenesis is essential for tissue regeneration and repair. A growing body of evidence shows that the use of bioactive glasses (BG) in biomaterial-based tissue engineering (TE) strategies may improve angiogenesis and induce increased vascularization in TE constructs. This work investigated the effect of adding nano-sized BG particles (n-BG) on the angiogenic properties of bovine type I collagen/n-BG composites. Nano-sized (20-30 nm) BG particles of nominally 45S5 Bioglass® composition were used to prepare composite films, which were characterized by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The in vivo angiogenic response was evaluated using the quail chorioallantoic membrane (CAM) as an model of angiogenesis. At 24 h post-implantation, 10 wt% n-BG containing collagen films stimulated angiogenesis by increasing by 41 % the number of blood vessels branch points. In contrast, composite films containing 20 wt% n-BG were found to inhibit angiogenesis. This experimental study provides the first evidence that addition of a limited concentration of n-BG (10 wt%) to collagen films induces an early angiogenic response making selected collagen/n-BG composites attractive matrices for tissue engineering and regenerative medicine.

Histologic and histomorphometric study of bone repair under acute Trypanosoma cruzi infection in rats.

Trypanosoma cruzi (T cruzi) is an intracellular protozoan pathogen that causes American trypanosomiasis (Chagas disease). The aim of this study was to evaluate the histopathological effects of acute infection by T. cruzi on bone repair, Wistar rats were used throughout. The animals were assigned to two groups: Control Group (CG n =20) and Experimental Group (EG n = 20). All the animals were anesthetized, at to the first lower right molar was extracted. The EG animals were inoculated subcutaneously at to with 0.1 mL of 10 trypomastigotes of the virulent strain Tulahuen of T. cruzi. The CG animals were administered an equivalent volume ofsaline solution subcutaneously. The animals in both groups were euthanized at 15 days post-infection and tooth extraction. The mandibles were resected, fixed informalin solution, radiographed, decalcified and embedded in paraffin. Bucco-lingually oriented sections were obtained at the level of the mesial tooth socket of the first lower molar and stained with hematoxylin-eosin. Total alveolar volume (TV) and bone volume (TBV/TV) in the apical third of the tooth socket were evaluated histomorphometrically. The histological analysis revealed an alteration in post-extraction bone tissue repair in animals infected by T. cruzi. A reduction in osteogenic activity was observed concomitant with a rise in quiescent and eroded bone surfaces. Histomorphometric evaluation revealed a significant reduction (19%) in total alveolar volume (TV) and bone volume (TBV/TV) (24%) in the apical third of the tooth socket in animals infected with T. cruzi in comparison to non-infected animals (p<0.05). The results obtained using this experimental model showed decreased osteogenesis in bone tissue repair under acute Trypanosoma cruzi infection in rats.

Histologic and histomorphometric study of bone repair under acute Trypanosoma cruzi infection in rats

Trypanosoma cruzi (T. cruzi) is an intracellular protozoan pathogen that causes American trypanosomiasis (Chagas disease). The aim of this study was to evaluate the histopathological effects of acute infection by T. cruzi on bone repair. Wistar rats were used throughout. The animals were assigned to two groups: Control Group (CG n =20) and Experimental Group (EG n =20). All the animals were anesthetized, at t0 the first lower right molar was extracted. The EG animals were inoculated subcutaneously at t0 with 0.1 mL of 105 trypomastigotes of the virulent strain Tulahuen of T. cruzi. The CG animals were administered an equivalent volume of saline solution subcutaneously. The animals in both groups were euthanized at 15 days post-infection and tooth extraction. The mandibles were resected, fixed in formalin solution, radiographed, decalcified and embedded in paraffin. Bucco-lingually oriented sections were obtained at the level of the mesial tooth socket of the first lower molar, and stained with hematoxylin-eosin. Total alveolar volume (TV) and bone volume (TBV/TV) in the apical third of the tooth socket were evaluated histomorphometrically. The histological analysis revealed an alteration in post-extraction bone tissue repair in animals infected by T. cruzi. A reduction in osteogenic activity was observed concomitant with a rise in quiescent and eroded bone surfaces. Histomorphometric evaluation revealed a significant reduction (19%) in total alveolar volume (TV) and bone volume (TBV/TV) (24%) in the apical third of the tooth socket in animals infected with T. cruzi in comparison to non-infected animals (p<0.05). The results obtained using this experimental model showed decreased osteogenesis in bone tissue repair under acute Trypanosoma cruzi infection in rats.

El Trypanosoma cruzi (T. cruzi) es un protozoario intracelular que causa Trypanosomoniasis Americana (Enfermedad de Chagas). El objetivo del presente trabajo fue el estudio histopatologico del efecto de la infeccion aguda por Trypanosoma cruzi sobre la reparacion del tejido oseo. Se utilizaron ratas Wistar macho que fueron asignadas a dos grupos: Grupo Control (GC n =20) y Grupo Experimental (GE n =20). Los animales de ambos grupos, bajo anestesia general intraperitoneal, fueron sometidos a t0, a exodoncia del primer molar inferior derecho, en el GE fueron inoculados,a t0 por via subcutanea en la region inguinal izquierda con 0.1 mL de 105 tripomastigotes de la cepa virulenta Tulahuen de Trypanosoma cruzi. A los animales del GC se les administro el volumen equivalente de solucion salina por via subcutanea. A los animales de ambos grupos se les practico la eutanasia a los 15 dias. Se resecaron las mandibulas, se fijaron en solucion de formol al 10%, se radiografiaron, se descalcificaron y se incluyeron en parafina. Se obtuvieron cortes orientados en sentido vestibulo-lingual a nivel del alveolo mesial del primer molar inferior derecho y se colorearon con hematoxilina-eosina para su posterior estudio histologico e histomorfometrico. Histologicamente se observo una menor actividad osteogenica a expensas de un incremento de las superficies quiescentes y de las superficies erosivas en el GE. En la evaluacion histomorfometrica se detecto disminucion estadiasticamente significativa del volumen oseo total (19%) y del volumen trabecular en el tercio apical del alveolo (24%) en el GE con respecto al GC (p<0.05). Los resultados obtenidos en este modelo experimental evidencian una disminucion de la osteogenesis en la reparacion osea en ratas con infeccion aguda por Trypanosoma cruzi.

Imaging resin-cast osteocyte lacuno-canalicular system at bone-bioactive glass interface by scanning electron microscopy.

The morphology of the osteocyte lacuno-canalicular system at the bone-biomaterial implant-interface has not been fully investigated. In this study, the resin-cast scanning electron microscopy technique was used, for the first time, to image the lacuno-canalicular network within neoformed bone around bioactive glass (BG) particles implanted in rat tibia bone marrow. The most salient finding was that the osteocyte canaliculi pass through the calcium-phosphorus layer formed at the bone-BG interface and reach the silica-rich layer of the reacted BG.

Histomorphometric and microchemical characterization of maturing dental enamel in rats fed a boron-deficient diet.

Few reports are available in the literature on enamel formation under nutritional deficiencies. Thus, we performed a study to determine the effects of boron (B) deficiency on the maturing dental enamel, employing the rat continuously erupting incisor as the experimental model. Male Wistar rats, 21 days old, were used throughout. They were divided into two groups, each containing ten animals: +B (adequate; 3-mg B/kg diet) and -B (boron deficient; 0.07-mg B/kg diet). The animals were maintained on their respective diets for 14 days and then euthanized. The mandibles were resected, fixed, and processed for embedding in paraffin and/or methyl methacrylate. Oriented histological sections of the continuously erupting incisor were obtained at the level of the mesial root of the first molar, allowing access to the maturation zone of the developing enamel. Dietary treatment did not affect food intake and body weight. Histomorphometric evaluation using undecalcified sections showed a reduction in enamel thickness (hypoplasia), whereas microchemical characterization by energy-dispersive X-ray spectrometry did not reveal alterations in enamel mineralization.

Effect of bioactive glasses on angiogenesis: a review of in vitro and in vivo evidences.

The incorporation of bioactive glass into bone tissue-engineered scaffolds can be widely beneficial based on emerging evidence in the literature about the angiogenic potential of this material, particularly 45S5 Bioglass((R)). This article reviews the literature discussing in vitro studies which have demonstrated that increases in angiogenic indicators have been achieved through both direct and indirect contact of relevant cells with 45S5 Bioglass((R)) particles or with their dissolution products. A few available in vivo studies confirming the ability of bioactive glass, incorporated into scaffolds, to stimulate neovascularization are also discussed. Suggestions for further research are given, highlighting the need for specific investigations designed to assess the effect of particular ion dissolution products from bioactive glasses and their relative concentration on angiogenesis both in vitro and in vivo.

Osteoconductivity of strontium-doped bioactive glass particles: a histomorphometric study in rats.

There is accumulating evidence that strontium (Sr)-containing bioceramics have positive effects on bone tissue repair. The aims of the present study were to evaluate the osteoconductivity of Sr-doped bioactive glass (BG) particles implanted in rat tibia bone marrow, and characterize the neoformed bone tissue by SEM-energy-dispersive X-ray microanalysis. Melt-derived BGs were prepared from a base 45S5 BG. Sr-doped glass (45S5.6Sr) was prepared using 6 wt % SrO as a substitute for the CaO. Histological analysis using undecalcified sections showed that new lamellar bone had formed along the surface of both 45S5 and 45S5.6Sr BG particles within 4 weeks. To evaluate osteoconductivity, affinity indices were calculated. At 30 days after implantation, 45S5 and 45S5.6Sr BGs had almost identical affinity indices (88% +/- 7% and 87% +/- 9%; p > 0.05). Strontium was not detected in the neoformed bone tissue surrounding 45S5.6Sr BG particles. These results indicate that 45S5.6Sr BG particles are osteoconductive when implanted inside the intramedullary canal of rat tibiae, and no alterations in bone mineralization, in terms of Ca/P ratio, were observed in the neoformed bone tissue around 45S5.6Sr BG particles.

Histomorphometric study of alveolar bone healing in rats fed a boron-deficient diet.

Bone healing after tooth extraction in rats is a suitable experimental model to study bone formation. Thus, we performed a study to determine the effects of boron (B) deficiency on bone healing by using this model. The first lower right molar of weanling Wistar rats was extracted under anesthesia. The animals were divided into two groups: +B (adequate; 3 mg B/kg diet), and -B (boron-deficient; 0.07 mg/kg diet). The animals in both groups were killed in groups of 10 at 7 and 14 days after surgery. The guidelines of the NIH for the care and use of laboratory animals were observed. The mandibles were resected, fixed, decalcified, and embedded in paraffin. Buccolingually oriented sections were obtained at the level of the mesial alveolus and used for histometric evaluations. Total alveolar volume (TAV) and trabecular bone volume per total volume (BV/TV) in the apical third of the alveolus were determined. Percentages of osteoblast surface (ObS), eroded surface (ES), and quiescent surface (QS) were determined. No statistical significant differences in food intake and body weight were observed. Histomorphometric evaluation found -B rats had 36% and 63% reductions in BV/TV at 7 and 14 days, respectively. When compared with +B rats, -B rats had significant reductions (57% and 87%) in ObS concomitantly with increases (120% and 126%) in QS at 7 and 14 days, respectively. The findings show that boron deficiency results in altered bone healing because of a marked reduction in osteogenesis.

A histomorphometric study of alveolar bone modelling and remodelling in mice fed a boron-deficient diet.

OBJECTIVE: Emerging evidence indicates that boron (B) plays a role in bone formation and maintenance. Thus, a study was performed to determine whether dietary B-deficiency affects periodontal alveolar bone modelling and remodelling. DESIGN: Weanling Swiss mice (n=30) were divided into three groups: control diet (GI, 3mg B/kg); B-deficient diet (GII, 0.07 mg B/kg); and pair-fed with GII (GIII). The animals were maintained on their respective diets for 9 weeks and then sacrificed. The guidelines of the NIH for the care and use of laboratory animals were observed. The mandibles were resected, fixed, decalcified in 10% EDTA and embedded in paraffin. Buccolingually oriented sections were obtained at the level of the mesial root of the first lower molar and stained with H-E. Histomorphometric studies were performed separately on the buccal and lingual sides of the periodontal alveolar bone. Percentages of osteoblast surfaces (ObSs), eroded surfaces (ESs), and quiescent surfaces (QSs) were determined. RESULTS: No statistically significant differences in food intake and body weight were observed between the groups. When compared with GI and GIII mice, GII mice (B-deficient) had 63% and 48% reductions in ObS and 58% and 73% increases in QS in buccal and lingual plates, respectively. ES were not affected by B nutriture. CONCLUSION: The results are evidence that dietary boron deprivation in mice alters periodontal alveolar bone modelling and remodelling by inhibiting bone formation.

Effects of intraosseous implantation of silica-based bioactive glass particles on rat kidney under experimental renal failure.

The aim of the present study is to evaluate the effects of intraosseous implantation of silica-based bioactive glass (BG) particles on rat kidney under experimental renal failure. The animals are assigned to one of the two groups: renal failure (RF) and renal failure + bioactive glass (RF + BG). Particles of melt-derived 45S5 BG are implanted in the marrow of one tibia of each animal in the RF + BG group. The animals are killed 24 h and 14 days postimplantation. The RF + BG group exhibits a statistically significant increase in serum urea 24 h postimplantation. The tibiae of the RF + BG group are resected and embedded in methyl-methacrylate resin. Ground sections are analyzed by light microscopy and energy-dispersive X-ray (EDX) analysis. The presence of silicon, calcium, and phosphorus is evaluated in the BG particles. A 55% reduction in silicon content is observed at 14 days postimplantation as compared with that at 24 h.Light microscopy analysis reveals lesions in kidney parenchyma. Hyperplasia associated with nuclear vacuolization in the tubules and a marked thickening of the basal membrane are observed in the renal cortex of the RF + BG animals killed at 24 h postimplantation, but not in those at 14 days. The present results demonstrate reversible renal cell injury in rats exposed to intraosseous implantation of silica-based BG particles under experimental RF.

A novel methodology for imaging new bone formation around bioceramic bone substitutes.

In this study, Frost's bulk-staining in combination with confocal laser scanning microscopy (CLSM) was used to image and characterize ground sections of undecalcified rat bone tissue with in situ bioceramic implants. This was addressed by bulk staining specimens in alcohol-soluble basic fuchsin dye. The ground sections were imaged using CLSM in the confocal fluorescence mode. Confocal images revealed that the newly formed bone could be clearly distinguished from bone marrow and cortical bone, as well as from the implant material.

A histomorphometric study of alveolar bone modeling and remodeling under experimental anaemia and polycythaemia in rats.

UNLABELLED: Alveolar bone is the least stable of the periodontal tissues, because it is subjected to continuous modeling and remodeling. OBJECTIVE: To perform a histological and histomorphometric evaluation of bone modeling and remodeling of periodontal alveolar bone under experimental anaemia and polycythaemia. METHODS: Thirty Wistar rats were divided into three groups: control (C), animals were i.p. injected with 0.5 mL of saline solution; anaemia (A), animals were injected with 6 mg/100 b.w. of phenylhydrazine every 48 h; polycythaemia (P), animals were transfused with 2.5 mL/100 b.w. of 80% suspension of homologous erythrocytes. All the animals were sacrificed 14 days after the onset of the experiment. The mandibles were resected, fixed in formalin, radiographed, processed and embedded in paraffin. Bucco-lingually oriented sections were obtained at the level of the mesial root of the first lower molar, and stained with hematoxylin-eosin. Histological and histomorphometric studies were performed on the buccal and lingual plates of periodontal alveolar bone. RESULTS: Histological and histomorphometric studies showed a statistically significant decrease in bone formation both in buccal and lingual plates in group A (anaemia) as compared to group C (control). An increase in active bone formation was found in the lingual plate in group P (polycythaemia) as compared to group C (control). CONCLUSION: The results obtained using this experimental model evidenced alterations in bone modeling and remodeling under conditions of anaemia and polycythaemia and/or associated factors.

Biological performance of boron-modified bioactive glass particles implanted in rat tibia bone marrow.

The aim of the present study was to characterize the neoformed bone tissue around boron-modified bioactive glass particles implanted in rat tibia bone marrow by histologic, histomorphometric and microchemical evaluation. Melt-derived glasses were prepared from a base 45S5 bioactive glass of nominal composition (45% SiO(2), 24.5% CaO, 24.5% Na(2)O and 6% P(2)O(5) in wt%). The glass composition was modified by adding 2% wt of boron oxide (45S5.2B). Histological and histomorphometric analyses using undecalcified sections showed that at 15 days post-implantation the area of neoformed bone tissue around the 45S5.2B particles was significantly higher than control 45S5 glass. No statistically significant differences were observed at 30 days post-implantation. The thickness of osseointegrated tissue on 45S5.2B BG particles was significantly greater than on the control at all experimental time-points evaluated. A statistically significant increase in the Ca:P ratio was observed in the neoformed bone around 45S5.2B particles 15 days post-implantation. The results of the present study provide evidence that particles of boron-modified 45S5 BG (45S5.2B) enhance bone formation more than 45S5 glass when implanted into the intramedullary canal of rat tibiae.

Mandibular bone remodeling under a choline-deficient diet: a histomorphometric study in rats.

BACKGROUND: A deficiency of lipotropic factors in the rat induces renal, hepatic, and/or hematic damage. The aim of the present study was to evaluate the effect of a choline-deficient diet and refeeding on mandibular bone remodeling. METHODS: Fifty Wistar rats were divided into 5 groups: group 1 (G1): control diet for 15 days; group 2 (G2): choline-deficient diet for 15 days; group 3 (G3): control diet for 30 days; group 4 (G4): choline-deficient diet for 30 days; and group 5 (G5): choline-deficient diet for 15 days and control diet for 15 days. All animals were sacrificed by ether overdose. The mandibles were resected, radiographed, decalcified, processed, and embedded in paraffin. Bucco-lingually oriented sections were obtained at the level of the interradicular bone of the medial roots of the left first molar, and stained with hematoxylin and eosin (H & E). Bone tissue density and bone remodeling were determined histomorphometrically. Body weight, food intake, hematocrit, and hemoglobinemia were also recorded. RESULTS: Microscopic observation revealed that osteogenesis was lower in rats fed a choline-deficient diet, at both 15 and 30 days, and that this decrease did not revert with a control diet. Histomorphometric evaluation showed 37% and 27% reduction in bone tissue density at 15 and 30 days, respectively, and a 30% decrease in bone formation at 30 days, compared to controls. CONCLUSION: In this experimental model, a choline-deficient diet led to altered bone remodeling as observed by a marked reduction in osteogenesis.