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.

Comparison of techniques for tibial dyschondroplasia assessment in broiler chickens

This study was carried out at Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual Paulista, Botucatu, SP, Brazil, with the objective of evaluating the efficacy of four techniques in the assessment of tibial dyschondroplasia lesions in broiler chickens. Four hundred Cobb male chicks were reared from 1 to 39 days of age. At 39 days, forty birds were selected and tibial dyschondroplasia status was assessed by four different techniques: evaluation using the lixiscope, macroscopic examination, histological examination and bone mineral density assessment using optical radiographic densitometry. The efficacy of each technique to assess dyschondroplasia lesions in the tibial growth plate was determined in comparison to histology, which was considered to be 100% efficient. The correlation results between lixiscope analysis and histology were poor. Macroscopic scores and densitometry readings were highly correlated with histology scores, and it is considered that these techniques reliably reproduce the status of the growth plate.

Evaluation of a radiographic method to detect tibial dyschondroplasia lesions in broilers

This study aimed at developing radiographic techniques for the early detection of dyschondroplastic lesions in the tibia of broilers. The experiment was carried out at the facilities of UNIFOR/MG and Formiga and UNIFENAS/ Alfenas with 420 one-day-old male Cobb broilers. At 20 days of age, all birds were radiographed and identified with an alpha-numeric metal ring in the right leg. At 40 days of age, 42 broilers previously selected as a function of bone mineral density and lesion thickness scores were again radiographed and scored, and then sacrificed. Their right tibia was removed for gross and histological examination of the growth plate. The results showed that radiographic techniques are correlated with gross and histological examination and that there was no significant differences among techniques (P>0.05). it was concluded that the use of radiographic examination to identify tibial dyschondroplasia in broilers precludes the use of bone mineral density to diagnose this condition. The non-parametric statistical Chi-square test at 5% significance level was used to analyze the results.

Evaluation of tibial dyschondroplasia development in broiler chickens fed diets containing 25 hydroxicolecalciferol

This research was carried out to evaluate the development of tibial dyschondroplasia (TD) in broilers from 1 to 21 days. Four hundred forty day-old male chickens, from broiler breeders aged 60-62 weeks, were randomly distributed in a 3×3+2 factorial arrangement, resulting in 11 treatments with four replicates of 10 birds each. The factors evaluated were: bird strains (Ross 308, Cobb 500, and Hybro), levels and sources of vitamin D (1250UI D3/kg without 25-(OH)D3; 1250UI D3/kg with 69mg 25-(OH)D3/ton, and 3000UI D3/kg with 69mg 25-(OH)D3/ton), and two control treatments containing calcium and phosphorus levels with 3000UI D3/kg without 25-(OH)D3 and 3000UI D3/kg with 69mg 25- (OH)D3/ton of feed. Bone characteristics were evaluated. From 1 to 21 days, the results showed that bone calcium and phosphorus concentrations were not influenced by the treatments, however, feed intake was higher for factorial than control treatments. Bone breaking resistance was not influenced by the treatments and there were no typical lesions of TD. In conclusion, it was not observed TD development in broilers from 1 to 21 days according to experimental procedures of this research.

Discondroplasia tibial: mecanismos de lesão e controle

A discondroplasia tibial (DT) é atribuída a uma assincronia no processo de diferenciação dos condrócitos, levando à formação de uma camada de condrócitos pré-hipertróficos e de uma cartilagem na tíbia proximal que não é calcificada, mas é resistente à invasão vascular. Além disso, tem sido proposto que, na discondroplasia tíbial, a etapa final do processo de calcificação não ocorre devido ao fato de que os efetores de alguns genes, relacionados com o mecanismo de calcificação do disco de crescimento podem apresentar algumas de suas propriedades químicas ou biológicas alteradas e/ou não serem expressos. Nesse sentido, a compreensão do mecanismo de ação e o papel das biomoléculas e dos minerais relacionados com a discondroplasia tibial poderão contribuir para o conhecimento de doenças do tecido ósseo e estabelecer estratégias de prevenção e tratamento.

Estudo da atividade das enzimas envolvidas na ossificação de frangos de corte normais e acometidos por discondroplasia tibial

Pós-graduação em Biotecnologia - IQ

Parâmetros biofísicos e morfométricos dos ossos, músculos e cartilagens de frangos de corte saudáveis e com discondroplasia tibial

Pós-graduação em Zootecnia - FCAV

Expressão das proteínas da matriz na discondroplasia da tíbia

A discondroplasia da tíbia (TD) em aves consiste numa anomalia do esqueleto onde existe uma falha nos processos normais da ossificação endocondral. Esta patologia é caracterizada pela formação de uma cartilagem não vascularizada e não mineralizada que se estende até à metáfise. Uma vez que existem várias anomalias do esqueleto em mamíferos com lesões semelhantes às apresentadas pela TD, este trabalho teve como objectivo a caracterização desta patologia em termos das moléculas que podem estar envolvidas no seu desenvolvimento. Assim, foi estudada a expressão das macromoléculas da matriz extracelular, das enzimas degradadoras da matriz (metaloproteinases da matriz: MMPs), bem como das moléculas envolvidas na proliferação e diferenciação celular, na angiogénese e apoptose. A expressão génica foi realizada, por PCR quantitativo em tempo real, em placas de crescimento normais e discondroplásicas obtidas a partir de frangos de carne (broilers) da estirpe Cobb. Os níveis proteicos de algumas MMPs foram analisados por immunoblotting e zimografia de gelatina. No presente estudo não se verificou alteração na expressão dos genes dos colagénios do tipo II, IX, X e XI, bem como do agrecano, nas lesões discondroplásicas. Observou-se uma redução acentuada nos níveis de mRNA da gelatinase-B (MMP-9), da colagenase-3 (MMP-13) e das estromalisinas -2 (MMP-10) e -3 (MMP-11), bem como nos níveis proteicos da gelatinase-A (MMP-2) e da MMP-13. Por outro lado, a MMP-7 aumentou drasticamente a expressão do seu gene. As moléculas envolvidas na proliferação e diferenciação dos condrócitos, tais como a PTHrP, o Ihh, o Cbfa-1 e o Sox-9, mantiveram a sua expressão génica nas lesões. Por outro lado, o TGF-β reduziu a sua expressão. A caspase-3 também dimimuiu a sua expressão na patologia. Em relação aos factores angiogénicos, o FGF manteve a sua expressão e o VEGF aumentou significativamente nas lesões. Este aumento do VEGF juntamente com o aumento da MMP-7 sugere um aumento da hipoxia nas lesões. Os nossos resultados sugerem que a acumulação da cartilagem observada na discondroplasia é devida a uma diminuição da proteólise da matriz, resultado de uma sub-expressão das MMPs, e não de um aumento da produção das macromoléculas da matriz. Desta forma, os nossos resultados sugerem que a falha na expressão e/ou activação das MMPs poderá estar associada ao desenvolvimento da discondroplasia da tíbia em aves. Finalmente, os nossos resultados vêm suportar os resultados anteriores que sugerem uma ligação entre a expressão das MMPs e anomalias no processo de ossificação endocondral.

Selecting appropriate bedding to reduce locomotion problems in broilers

Two experiments were carried out at the Poultry Sector of the School of Agrarian Sciences of the Federal University of Grande Dourados to evaluate the incidence of leg problems in broilers reared on two distinct types of bedding material: rice husks or wood shavings, both new and reused. In both trials, a randomized experimental design was applied in factorial arrangement (2 x 2 x 2) using two genetic strains (Cobb® or Ross®); two sexes (male or female), and two litter materials (rice husks or wood shavings). In each trial 1080 one day pullets were reared equally divided in the treatments. The birds were placed in 4.5 m² boxes at a density of 10 birds m-2. All birds were fed diets with equal nutritional density, and water was offered ad libitum. Feeds were divided in three phases: starter diet (1 - 21 days), grower diet (22 - 35 days), and finisher diet (36 - 45 days). on day 45, fifty birds were randomly selected in each experiment to evaluate flock leg problems. The following parameters were analyzed: gait score, incidence of valgus and varus disorder, footpad dermatitis, femoral degeneration, tibial dyschondroplasia, and spondylolisthesis. Ambient temperature during rearing and litter caking and moisture content were recorded in four boxes per treatment. The analytical hierarchy process was used to organize the data into specific criteria. Several criteria, related to the attributes that were determinant according to the statistical analysis, were chosen in order to provide the best input to the process. Results indicated that new wood-shavings bedding was the most appropriate bedding to prevent locomotion problems, followed by new rice husks, reused wood shavings, and reused rice husks. However, when leg problems were associated to sex and genetic strain, male Ross birds strain presented less problems when reared on new rice husks, followed by new wood shavings

Use of vitamin D to reduce lameness in broilers reared in harsh environments

In tropical poultry-producing countries, poultry houses usually have little environmental control. This study investigated the effect of dietary vitamin D on the incidence of leg abnormalities of a fast-growing broiler strain reared under harsh conditions. In this study, 300 one-day-old male broilers were distributed in two treatments with three replicates of 50 birds each. One group was fed a placebo and the other group was fed 25-hydroxycholecalciferol (25-OH-D3) in a soluble form. The environmental variables were weekly recorded during until birds were 49 days old. Birds were weekly gait scored, and their walking speed and vertical force while walking were assessed. Post-mortem examination was performed and skeletal lesions were assessed. Control birds presented more leg problems than those that ingested 25-OH-D3. These results suggest that dietary vitamin D is effective in decreasing the severity of lameness by reducing tibial dyschondroplasia and other leg abnormalities.

Locomotion problems of broiler chickens and its relationship with the gait score

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

Sodium and chloride requirements of young broiler chickens fed corn-soybean diets (one to twenty-one days of age)

Sodium (Na+) and chloride (Cl-) nutritional requirements, dietary electrolyte balance (DEB), and their effects on acid-base balance, litter moisture, and tibial dyschondroplasia (TD) incidence for young broiler chickens were evaluated in two trials. One-day-old Cobb broilers were distributed in a completely randomized design with six treatments, five replicates, and 50 birds per experimental unit. Treatments used in both experiments were a basal diet with 0.10% Na+ (Experiment 1) or Cl- (Experiment 2) supplemented to result in diets with Na+ or Cl- levels of 0.10, 0.15, 0.20, 0.25 ,0.30, or 0.35%, respectively. In Experiment 1, results indicated an optimum Na+ requirement of 0.26%. Sodium levels caused a linear increase in arterial blood gas parameters, indicating an alkalogenic effect of Na+. The hypertrophic area of growth plate in the proximal tibiotarsi decreased with Na+ levels. The TD incidence decreased with increases in dietary Na+. Litter moisture increased linearly with sodium levels. In Experiment 2, the Cl- requirement was estimated as 0.25%. Chloride levels caused a quadratic effect (P ≤ 0.01) on blood gas parameters, with an estimated equilibrium [blood base excess (BE) = 0] at 0.30% of dietary CT-. No Cl- treatment effects (P ≥ 0.05) were observed on litter moisture or TD incidence. The best DEB for maximum performance was 298 to 315 mEq/kg in Experiment 1 and 246 to 264 mEq/kg in Experiment 2. We concluded that the Na+ and Cl- requirements for optimum performance of young broiler chickens were 0.28 and 0.25%, respectively.

Morfologia do tecido ósseo de tíbias de frangos de corte submetidos à dieta com 25 hidrocolecalciferol

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Establishment of a preclinical ovine model for tibial segmental bone defect repair by applying bone tissue engineering strategies

Currently, well-established clinical therapeutic approaches for bone reconstruction are restricted to the transplantation of autografts and allografts, and the implantation of metal devices or ceramic-based implants to assist bone regeneration. Bone grafts possess osteoconductive and osteoinductive properties, however they are limited in access and availability and associated with donor site morbidity, haemorrhage, risk of infection, insufficient transplant integration, graft devitalisation, and subsequent resorption resulting in decreased mechanical stability. As a result, recent research focuses on the development of alternative therapeutic concepts. The field of tissue engineering has emerged as an important approach to bone regeneration. However, bench to bedside translations are still infrequent as the process towards approval by regulatory bodies is protracted and costly, requiring both comprehensive in vitro and in vivo studies. The subsequent gap between research and clinical translation, hence commercialization, is referred to as the ‘Valley of Death’ and describes a large number of projects and/or ventures that are ceased due to a lack of funding during the transition from product/technology development to regulatory approval and subsequently commercialization. One of the greatest difficulties in bridging the Valley of Death is to develop good manufacturing processes (GMP) and scalable designs and to apply these in pre-clinical studies. In this article, we describe part of the rationale and road map of how our multidisciplinary research team has approached the first steps to translate orthopaedic bone engineering from bench to bedside byestablishing a pre-clinical ovine critical-sized tibial segmental bone defect model and discuss our preliminary data relating to this decisive step.

CYR61 (CCN1) protein expression during fracture healing in an ovine tibial model and its relation to the mechanical fixation stability

The formation of new blood vessels is a prerequisite for bone healing. CYR61 (CCN1), an extracellular matrix-associated signaling protein, is a potent stimulator of angiogenesis and mesenchymal stem cell expansion and differentiation. A recent study showed that CYR61 is expressed during fracture healing and suggested that CYR61 plays a significant role in cartilage and bone formation. The hypothesis of the present study was that decreased fixation stability, which leads to a delay in healing, would lead to reduced CYR61 protein expression in fracture callus. The aim of the study was to quantitatively analyze CYR61 protein expression, vascularization, and tissue differentiation in the osteotomy gap and relate to the mechanical fixation stability during the course of healing. A mid-shaft osteotomy of the tibia was performed in two groups of sheep and stabilized with either a rigid or semirigid external fixator, each allowing different amounts of interfragmentary movement. The sheep were sacrificed at 2, 3, 6, and 9 weeks postoperatively. The tibiae were tested biomechanically and histological sections from the callus were analyzed immunohistochemically with regard to CYR61 protein expression and vascularization. Expression of CYR61 protein was upregulated at the early phase of fracture healing (2 weeks), decreasing over the healing time. Decreased fixation stability was associated with a reduced upregulation of the CYR61 protein expression and a reduced vascularization at 2 weeks leading to a slower healing. The maximum cartilage callus fraction in both groups was reached at 3 weeks. However, the semirigid fixator group showed a significantly lower CYR61 immunoreactivity in cartilage than the rigid fixator group at this time point. The fraction of cartilage in the semirigid fixator group was not replaced by bone as quickly as in the rigid fixator group leading to an inferior histological and mechanical callus quality at 6 weeks and therefore to a slower healing. The results supply further evidence that CYR61 may serve as an important regulator of bone healing.