Bone mineral density of tibae and femura of broiler breeders: growth, development and production

The aim of this study was to follow-up the physiological variations in the development of the bone tissue, associating them with the egg production curve. This study was carried out in the facilities of the Faculdade de Medicina Veterinária e Zootecnia of the UNESP, Botucatu, Brazil. Twenty-three families of Ross broiler breeders were used, each family consisting of 13 females and 1 male, distributed in 23 pens of 5.0m² each. The management was that recommended by the genetic company manual (Agroceres Ross, 2003), with daily feeding until 6th week of age; and birds were fed according to a 5:2 schedule (5 days fed, 2 days of fasting) between 7 and 17 weeks of age, returning to daily feeding starting at 18 weeks of age. Birds did not receive afternoon calcium supplementation. on the fourth week of rearing, 84 females were removed for bone analyses of the right tibia and femur, using optical densitometry in radiographic images technique. These analyses were sequentially carried out in 4, 8, 12, 15, 20, 24, 30, 35, 42, 47, and 52 week-old birds. The egg production curve of the birds was followed-up and associated to bone mineral density results. For bone mineral density evaluation (BMD) birds were divided by weight categories as light, intermediate, or heavy within each data age. BMD values of the tibias were not influenced by weight range, but by the age at collection. on the other hand, interactions were found among femur BMD values and weight and age categories. There was no correlation between eggshell quality and femur BMD. A negative correlation (-0.15) was observed between tibia BMD and eggshell percentage. It was possible to conclude that the egg production has little influence on bone mineral density of the birds probably because there was no need of bone mineral mobilization during the production period, since the observed egg production was below that observed under commercial conditions.

Use of radiograph optical densitometry to monitor bone quality in broiler breeders

This study was carried out at Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual Paulista, Botucatu, SP, Brazil, and evaluated bone quality in broiler breeders. Twenty-three families of Ross broiler breeders were housed in 5.0-m² pens. The families were comprised of 13 females and one male at the onset of the experimental period. The mean number of females per family was 9.34 at the end of the trial. The feeding program and management followed strain guidelines (Agroceres Ross, 2003). Bone analyses were performed in the right tibia and femur using optical radiographic densitometry at 4, 8, 12, 15, 20, 24, 30, 35, 42, 47 and 52 weeks of rearing. Trap nests were used to collect eggs from the breeders two weeks before and after the evaluation weeks. At each evaluation day, five birds were sacrificed after radiographs were taken and the tibias and femurs were collected to perform the following analyses: fatfree dry matter, ash percentage, bone resistance and Seedor index. Therefore, it was possible to establish correlations between bone quality and eggshell quality. Characteristics of bone quality were highly correlated to each other; on the other hand, there were no correlations between bone quality and external egg quality. In conclusion, there was no effect of egg production on egg quality, possibly because there was no reabsorption of bone minerals.

Tibial dyschondroplasia and bone mineral density

This study was carried out at Faculdade de Medicina Veterinária e Zootecnia, UNESP, Botucatu-SP, Brazil. The aim was to establish the normal values of bone mineral density (BMD) expressed in millimeters of aluminum in the tibia of broiler chickens using optical densitometry of radiographs. Four hundred Cobb male chicks were reared from 1 to 40 days of age, when 40 of them were selected and the right femur-tibia articulation was radiographed. Radiographs were taken with the X-ray equipment calibrated for 45 kvp and 3.2 mAs and a focus-to-film distance of 90 cm. An aluminum phantom ASTM-6063 consisting of 20 ladder steps with graduate density was placed parallel to the area to be radiographed and used as a densitometry reference standard. Radiograph images were analyzed using the software ATHENA - SIA. The proximal growth plate of the right tibia epiphysis was used as the standard reading region. The inclination axis of the reading window was 0 and the window was 10 mm high and 40-45 mm wide, depending on the bone size. Optical densitometry values of the radiographs ranged from 1.46 to 1.77 mmAl, and the coefficient of variation was 9.93%. It was concluded that densitometry values beyond the range established in the present study might indicate the presence of bone alteration in the tibia of broilers.

Follow-up of the Development of Femoral Degeneration Lesions in Broilers

A study was carried out in the experimental facilities of FMVZ/UNESP-Botucatu, with the aim of following-up the development and the incidence of femoral degeneration (FD). A total of 305 one-day-old male broilers were housed in six pens of 5m(2) each. Histological analyses of femur head collected when broilers were 0, 7, 14, 21, 28, 35, and 42 days of age were carried out. At 42 days of age, 30 birds were taken to the experimental processing plant of FMVZ for leg gross examination. Ten legs per FD score where selected, and histologically analyzed to determine the most probable age at the beginning of the lesions, and to standardize femoral degeneration lesion scores. The histological results showed that cell architecture started to disorganize at 21 days of age in the resting and proliferation zones, and that angiogenesis increased, invading the joint cartilage, The gross lesion indexes due to femoral degeneration were 22.5%, 42.5%, and 65% at 28, 35, and 42 days of age, respectively.

A Modified Technique of Rat Tail Suspension for Longer Periods of Observation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Radiographic assessment of bone density around integrated titanium implants after ovariectomy in rats

Objectives:This study evaluated the influence of ovariectomy 8 weeks after implant placement on bone integrated to titanium implants.Materials and methods:Thirty-eight female rats were submitted to a titanium implant at the tibiae proximal methaphysis. After a healing period of 8 weeks the animals were randomly divided into three groups: control (CTL-10 animals), sham-operated (SHAM-14 animals) and ovariectomy (OVX-14 animals). The CTL group was then sacrificed in order to confirm integration of the implant. The SHAM group was submitted to fictitious surgery and the OVX group was submitted to bilateral ovariectomy. After 12 weeks post-implant placement, the SHAM and OVX groups were sacrificed. In order to confirm the systemic osteopenia in rats, a dual-energy X-ray absorptiometry (DXA) was performed. For the evaluation of bone density, digital radiographs were taken. The grey level of the bone adjacent to implant was measured using image software and the bone density was calculated at six points on both sides of the implant.Results:Densitometry measurements of the femur confirmed systemic bone mass loss in the OVX group. Individualized bone analyses of different regions surrounding the implant showed a significantly lower radiographic bone density (P < 0.05) in the cancellous region of the OVX group (77.48 +/- 23.39 grey levels) when compared with the CTL and SHAM groups (91.61 +/- 32.10 and 102.57 +/- 32.50 grey levels, respectively).Conclusions:The present study showed a decrease of the radiographic bone density in the cancellous region of bone around titanium implants placed 8 weeks before ovariectomy in rats.

The influence of loss of bone mass on induced periodontal disease: A radiographic and densitometric study of female rats

Background: Changes in mineral density in the mandibular and femoral bones (BMD) after estrogen deficiency caused by ovariectomy (OVX) and the influence of these changes on induced periodontal disease were evaluated in female rats.Methods: Forty-eight female Holtzman rats (90 days old) were randomly divided into five groups: 0: control (N = 9); 1: SHAM without induced periodontal disease (N = 11); 2: SHAM with induced disease (N = 10); 3: OVX without induced disease (N = 9); and 4: OVX with induced disease (N = 9). In groups 2 and 4, the first lower molars were tied with ligatures for 30 days 120 days after surgery. After 5 months the animals were sacrificed to measure global mineral density (BMD) and that of the sub-regions of the mandible and femur by dual energy x-ray absorptiometry (DXA). The extent of vertical bone loss was evaluated with digital radiography by measuring the distance from the bone crest to the cemento-enamel junction at the mesial of the first lower molar.Results: Results of the femur (Kruskal-Wallis test) showed a significant difference (P < 0.001) between the groups SHAM and OVX in bone density values for all regions. Comparison between the groups in relation to the BMD of the mandible, both in the sub-regions and global revealed no differences (P < 0.05). The vertical bone loss measured for the groups with induced disease was similar (P= 0.713).Conclusions: Differences between the groups were found in the bone mineral density BMD of the femur but not of the mandible. OVX had no influence on induced periodontal disease.

Effect of Severe Dietary Magnesium Deficiency on Systemic Bone Density and Removal Torque of Osseointegrated Implants

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The effect of oestrogen and alendronate therapies on postmenopausal bone loss around osseointegrated titanium implants

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Histological and three-dimensional evaluation of osseointegration to nanostructured calcium phosphate-coated implants

Nanostructures on implant surfaces have been shown to enhance osseointegration; however, commonly used evaluation techniques are probably not sufficiently sensitive to fully determine the effects of this process. This study aimed to observe the osseointegration properties of nanostructured calcium phosphate (CaP)-coated implants, by using a combination of three-dimensional imaging and conventional histology. Titanium implants were coated with stable CaP nanoparticles using an immersion technique followed by heat treatment. Uncoated implants were used as the control. After topographical and chemical characterizations, implants were inserted into the rabbit femur. After 2 and 4 weeks, the samples were retrieved for micro-computed tomography and histomorphometric evaluation. Scanning electron microscopy evaluation indicated that the implant surface was modified at the nanoscale by CaP to obtain surface textured with rod-shaped structures. Relative to the control, the bone-to-implant contact for the CaP-coated implant was significantly higher at 4 weeks after the implant surgery. Further, corresponding 3-D images showed active bone formation surrounding the implant. 3-D quantification and 2-D histology demonstrated statistical correlation; moreover, 3-D quantification indicated a statistical decrease in bone density in the non-coated control implant group between 2 and 4 weeks after the surgery. The application of 3-D evaluation further clarified the temporal characteristics and biological reaction of implants in bone. (C) 2011 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Low-level laser on femoral growth plate in rats

OBJETIVO: Determinar a influência do Laser Terapêutico de Baixa Potência sobre a placa de crescimento de ratos. MÉTODOS: Trinta ratos Wistar machos com 40 dias de idade foram divididos em dois grupos, G1 e G2. O grupo G1 foi submetido à irradiação com laser GaAlAs 830 nm, potência de saída de 40 mW, e densidade de energia de 10 J/cm2. A irradiação foi aplicada diariamente por um período máximo de 21 dias. O mesmo procedimento foi realizado no grupo G2, com a probe desativada. Cinco animais em cada grupo foram sacrificados nos dias 7, 14 e 21 e submetidas à análise histomorfométrica. RESULTADOS: em ambos os grupos, o disco fisário esteve radiograficamente visível em todos os momentos nas incidências craniocaudal e médio-lateral. No 21º dia a porcentagem de comprimento longitudinal do fêmur foi maior em G1 que em G2 em relação ao valor basal, e o número de condrócitos da zona hipertrófica foi maior em G1 que em G2. A zona de cartilagem calcificada estava maior em G1 em relação a G2 em todos os momentos de avaliação. A angiogênese foi maior em G1 que em G2 nos 14º e 21º dias. CONCLUSÃO: A terapia com laser terapêutico de baixa potência influenciou negativamente o disco fisário distal do fêmur de ratos.

Ectopic mineralization of articular cartilage in the bullfrog Rana catesbeiana and its possible involvement in bone closure

Mineralization of the articular cartilage is a pathological condition associated with age and certain joint diseases in humans and other mammals. In this work, we describe a physiological process of articular cartilage mineralization in bullfrogs. Articular cartilage of the proximal and distal ends of the femur and of the proximal end of the tibia-fibula was studied in animals of different ages. Mineralization of the articular cartilage was detected in animals at 1 month post-transformation. This mineralization, which appeared before the hypertrophic cartilage showed any calcium deposition, began at a restricted site in the lateral expansion of the cartilage and then progressed to other areas of the epiphyseal cartilage. Mineralized structures were identified by von Kossa's staining and by in vivo incorporation of calcein green. Element analysis showed that calcium crystals consisted of poorly crystalline hydroxyapatite. Mineralized matrix was initially spherical structures that generally coalesced after a certain size to occupy larger areas of the cartilage. Alkaline phosphatase activity was detected at the plasma membrane of nearby chondrocytes and in extracellular matrix. Apoptosis was detected by the TUNEL (TDT-mediated dUTP-biotin nick end-labeling) reaction in some articular chondrocytes from mineralized areas. The area occupied by calcium crystals increased significantly in older animals, especially in areas under compression. Ultrastructural analyses showed clusters of needle-like crystals in the extracellular matrix around the chondrocytes and large blocks of mineralized matrix. In 4-year-old animals, some lamellar bone (containing bone marrow) occurred in the same area as articular cartilage mineralization. These results show that the articular cartilage of R. catesbeiana undergoes precocious and progressive mineralization that is apparently stimulated by compressive forces. We suggest that this mineralization is involved in the closure of bone extremities, since mineralization appears to precede the formation of a rudimentary secondary center of ossification in older animals.

Efeitos do treinamento físico sobre a resistência mecânica do terço proximal do fêmur de ratos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Developmental exposure to diuron causes splenotoxicity in male Sprague-Dawley rat pups

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Characterization of mineral contents on androgen deficient and chronic ethanol consuming rats by biochemistry and non-destructive techniques

Ethanol can compromise the body mineral composition and affect bone, and when associated to hypogonadism is considered an important risk factor for osteoporosis in man. The aim of this study was to investigate the effect of androgen deficient and chronic ethanol consuming on mineral contents by biochemistry and non-destructive techniques. Wistar rat (n=54) were divided in orchiectomy (ORQ) or SHAM-operated and subdivided by diet. They were daily fed with a Lieber DeCarli diet model for 8 weeks long. The controls groups were free-diet and pair-fed. Ca and P were analyzed by biochemistry test in the blood and by nX-ray fluorescence and FT-Raman on the femur area. Serum analysis revealed hypocalcaemia and hypeiphosphataemia in ethanol groups more than pair-fed and free-diet. In similarity, spectroscopy indicated a decrease in bone Ca content in ORQ groups, mainly for ethanol groups. Phosphorus content and Ca/P molar ratio, otherwise, doesn't diverge in all 6 groups. Ethanol consumption impaired Ca and P homeostasis in ORQ rat more than SHAM. The relationships among ethanol consume and androgen deficit support the hypothesis that ethanol affects the mineral-regulating hormones and may mediate some effects on bone. These findings demonstrate that ethanol seemed to interfere with the normal compensatory response to these Ca and P levels and is more significant M androgen deficiency rats.