Longitudinal Growth of Leg and Wing Bones of Chickens Reared in Cages and on the Floor

The histomorphometry of proximal growth plate and rate of longitudinal growth was estimated in tibia and humerus of chickens reared in the cages and on the floor, during 0, 2, 4, 6 and 8 weeks of age. The floor vs. the cage area was 3:1. All chicks were fed ad libitum standard broilers mixture: starter (0-4 weeks) with 0.9% Ca; 0.7% P and grower (4-8 weeks) with 0.8% Ca; 0.5% P. The rearing system showed no significant effect on macroscopy estimated tibia and humerus growth in length, just as in case of bone's proximal growth plate microscopy of floor reared chickens. However, our results suggested that wing bones are more affected by the housing system than that of leg bones. The proliferative zone, hypertrophic zone and total growth plate of the humerus in cage reared birds were significantly higher than in those reared on the floor.     

硒对STZ诱发糖尿病大鼠骨密度影响的研究初报

比较了在低硒饲料的基础上补充不同形态硒对糖尿病大鼠骨密度的影响。健康SD大鼠 1 2 8只 ,体重 50～ 60 g,随机分为 4组 ,每组 32只 ,雌雄各半 ,其中Ⅰ组为低硒对照组 (饲料含硒量 0 0 37mg/kg) ;Ⅱ组为补充亚硒酸钠组 (饲料含硒量 0 3mg/kg) ;Ⅲ组为补充富硒麦芽组(饲料含硒量 0 3mg/kg) ;Ⅳ组为补充富硒酵母组 (饲料含硒量 0 3mg/kg)。Ⅱ、Ⅲ、Ⅳ组的饲料均是在Ⅰ组低硒饲料的基础上分别添加适量的亚硒酸钠、富硒麦芽、富硒酵母后配制而成。喂饲 5周后 ,Ⅰ、Ⅱ、Ⅲ、Ⅳ组用链脲佐菌素 (streptozotocin ,STZ)按 60mg/kg腹腔注射诱发糖尿病 ,继续饲喂 6周。用单光子吸收法测定各组大鼠的肱骨、股骨、胫骨、桡骨的骨密度。结果表明 :①STZ致糖尿病大鼠的骨密度随时间的延长有逐渐下降的趋势。Ⅱ、Ⅲ、Ⅳ补硒组大鼠可以在一定时间内延缓STZ致糖尿病大鼠的骨密度下降。在此期间骨密度的下降以肱骨出现最早 ,股骨次之 ,桡骨、胫骨再次之。②饲料补硒组糖尿病大鼠的肱骨、股骨中硒的含量较饲料低硒组糖尿病大鼠的肱骨、股骨中硒的含量显著提高。③补充硒的形态 ,即有机硒 (富硒麦芽、富硒酵母 )或无机硒 (亚硒酸钠 )对糖尿病大鼠肱骨、股骨、胫骨、桡骨的骨密度影响差异不显著。     

Histological disorders related to the focal disappearance of the epiphyseal growth plate in rats induced by high dose of vitamin A

The histological disorders related to the focal disappearance of the epiphyseal growth plate were examined histochemically in the proximal tibia of rats administered a high dose of vitamin A. Animals were given 100,000 IU/100 g body weight/day of vitamin A for 5 days from 4 weeks after birth (VA rats) or given deionized water as control and sacrificed on Day 12 and 19 of the experiment. Tibiae were examined by immunohistochemistry for type I, II and X collagens, lectin-histochemistry for Helix pomatia and backscattered electron imaging. On Day 12, the abnormally developed calcified cartilage matrix was detected within the epiphyseal growth plate in VA rats. The uncalcified cartilage matrix contained type I collagen but lacked type II collagen. In addition, the eroded regions accompanied with numerous osteoclasts and osteoblasts were detected in the epiphyseal growth plate. On day 19, eroded regions penetrated the epiphyseal growth plate to result in its focal disappearances with the eroded surfaces entirely covered with bone tissue in VA rats. These findings suggested that the cartilage matrix of the epiphyseal growth plate was abnormally calcified and showed the phenotypes like bone matrix. The eroded regions of the epiphyseal growth plate seemed to be caused by the invasion of osteoclasts into the altered cartilage matrix and might develop to the focal disappearances by the modeling or remodeling due to action of osteoclasts and osteoblasts.     

Healing of Canine Autogenous Cancellous Bone Graft Donor Sites

Autogenous cancellous bone was procured from the proximal ends of one humerus and the contralateral tibia of eight adult mixed-breed dogs. Greater weights of bone were consistently harvested from the humerus than from the tibia. Restoration of cancellous bone was more rapid and complete in the humerus than in the tibia. The tibia harvest site filled primarily with fibrous tissue rather than with cancellous bone.     

肉鸡胫骨干骺软骨的形态学观察

对3周龄肉鸡胫骨近端干骺软骨进行解剖学观察,发现干骺软骨内除了有骺软骨生长板外,还有次级骨化中心生长板。     

Percutaneous biopsy of the proximal humeral growth plate in dogs

A percutaneous biopsy technique for the study of endochondral bone formation in the dog was developed. With the dogs under general anesthesia or sedated with a combination of a tranquilizer and a local anesthetic, biopsy specimens were obtained from the proximal growth plate of the humerus with the use of a Jamshidi bone biopsy needle. Biopsy specimens were structurally intact, and contained epiphysis, growth plate, and metaphysis. The procedure proved to be a simple, safe technique, which caused minimal discomfort for the patient and did not affect the growth of the proximal end of the humerus, even after multiple biopsies.     

Distribution of cortical bone in bovine limbs

Xenogenic bone grafts have been widely researched because they are not limited in terms of volume and size like autogenous and allogenic grafts, and the favored bone for xenografts is bovine bone. However, the efficacy of cortical bone from bovine limb for xenografts is not clearly known. In this study, the distribution of cortical bone in bovine humerus, radius, femur, and tibia were investigated. Each experimental bone was split longitudinally with a bone saw and bone marrow and cancellous bone were removed. The thicknesses of cortical layers in sample diaphyses were measured at 1cm intervals using a micrometer. The mean lengths of cortical portions were; humerus 14.7 +/- 2.3 cm, radius 19.0 +/- 2.6 cm, femur 19.0 +/- 3.2 cm, and tibia 23.0 +/- 3.1 cm. Thickest cortical bone was found at the distal caudal metaphysis of the humerus, the proximal caudal metaphysis of the radius, the craniolateral and caudomedial midshaft of the femur and the lateral and medial midshaft of tibia. The mean surface areas of cortical bone were humerus 187.4 +/- 15.44 cm2, radius 229.2 +/- 43.31 cm2, femur 295.8 +/- 8.93 cm2, and tibia 290.0 +/- 30.44 cm2. And, mean volumes of cortical bone were humerus 149.42 +/- 15.35 cm3, radius 166.26 +/- 20.02 cm3, in femur 220.45 +/- 22.73 cm3, and tibia 214.89 +/- 20.05 cm3. The results of this study can be used to produce cortical bone-based plates and screws.     

Age-related changes in bone mineral density, cross-sectional area and the strength of long bones in the hind limbs and first lumbar vertebra in female Wistar rats

Age-related changes in bone mineral density (BMD) and cross-sectional area and bone strength index (SSI) of the femur, tibia, humerus, and first lumbar vertebra in female Wistar (WM/MsNrs) rats were examined by a quantitative computed tomography (pQCT) method. One hundred and sixteen virgin female Wistar (WM/MsNrs) rats aged 2-33 months were used. The data indicate that the total BMD values of metaphyses and diaphyses of long bones increased until 12 months, then decreased to a varying degree depending on the bone after 15-24 months, but the values of cortical and trabecular BMD with age were not always similar to the total BMD value. Nevertheless, the values for cross-sectional area and SSI in the long bones increased regardless of the total BMD decrease with age, indicating that this increase might have been due to a characteristic of the modeling pattern in rats. The total and cortical BMD values in the first lumbar vertebra decreased after 18 months, and SSI did after 15 months. The data obtained in this study were compared with those obtained from males in a previous study. In conclusion, it was indicated that in this strain the rats over 12 months with the highest total BMD values in the femur and tibia, and before the onset of various tumors, are useful as a model animal for osteoporosis experiments and observation of senile bone change.     

Bone structure and strength at different ages in laying hens and effects of dietary particulate limestone,vitamin K and ascorbic acid

1. A range of bone structural and strength characteristics was determined in laying hens at 15, 25, 50 and 70 weeks of age. The birds were fed up to 25 weeks on diets supplemented with additional vitamin K (10 mg menadione/kg) or ascorbic acid (250 mg/kg) or up to 70 weeks on diets containing limestone in powder or particulate form. 2. There were important effects of age on all bone characteristics. Between 15 and 25 weeks there was a rapid loss of cancellous bone and a rapid accumulation of medullary bone in the proximal tarsometatarsus (PTM). These changes continued at a slower rate up to 70 weeks. Cancellous bone content of the free thoracic vertebra (FTV) also declined after 15 weeks. 3. Breaking strengths of tibia and humerus did not change between 15 and 25 weeks but decreased later in lay. 4. None of the nutritional treatments affected bone characteristics at 15 weeks of age. 5. Increasing the dietary vitamin K supplement from 2 to 12 mg menadione/kg increased cancellous bone volume in the PTM at 25 weeks. 6. Dietary ascorbic acid did not affect any of the bone characteristics measured up to 25 weeks. 7. Particulate limestone resulted in a smaller loss of cancellous bone between 15 and 25 weeks and increased accumulation of medullary bone in the PTM. Breaking strength of the tibia and radiographic densities of tibia and keel were also improved. 8. It is concluded that patterns of bone loss over the lifetime of laying hens vary, depending upon the bone type. Feeding a particulate source of calcium can help to alleviate some of the characteristics of osteoporosis. Supplementation with extra vitamin K may also be beneficial.     

Differences in composition of avian bone collagen following genetic selection for resistance to osteoporosis

1. Collagen characteristics were compared in the tibiotarsus and humerus from 103 females and 38 males aged 68 to 72 weeks from the G6 generation of lines of laying hen selected for resistance or susceptibility to osteoporosis (high and low bone index (BI) lines). 2. Selection over the latest generation resulted in further divergence in the breaking strengths of humerus (from 12.3 to 21.8%) and tibia (from 22.3 to 37.3%) in hens. Males also showed line differences in bone strengths. 3. Plasma pyridinoline concentration was higher in hens in the low BI line, suggesting a greater rate of bone resorption in this line. 4. There were few differences between the lines in collagen and calcium concentrations in humerus and tibiotarsus cortical bone. 5. There were no differences between the lines in either sex in reduced immature collagen cross-link content of humerus or tibiotarsus. 6. Mature collagen cross-link content was higher in the high BI line in the male humerus but this effect was not apparent in the male tibiotarsus nor in either bone in the females. 7. Pyrrolic cross-link contents were higher in the high BI line in the female humerus and tibiotarsus and in the male tibiotarsus. 8. Over both lines combined, there were positive correlations between humeral and tibiotarsal pyrrole contents and strengths in females and between tibiotarsal pyrrole content and strength in males. 9. It is concluded that an increase in cross-linking, particularly pyrrolic cross-linking, in the collagen matrix contributes in part to the improvement in bone strength in the high BI line.     

Medial Plating for the Repair of Middle and Distal Diaphyseal Fractures of the Humerus in Dogs

Six middle and distal humeral shaft fractures in dogs were repaired by the application of a bone plate along the medial shaft of the humerus. In all animals, clinical and radiographic evidence of normal bone healing was observed. The bone plate and ancillary implants were removed from five dogs by 10 months. In none of the six dogs were any untoward effects of placing the bone plate along the medial shaft of the humerus detected nor were there any difficulties encountered with the medial humeral soft tissues.     

Evaluation of the proximal portion of the femur as an autogenous cancellous bone donor site in dogs.

The proximal portion of the femur was evaluated as a source of autogenous cancellous bone in dogs. Bilateral oval cortical defects were created in the lateral subtrochanteric area of the femur in 16 dogs. Cancellous bone was removed and the weight recorded. Cancellous bone was similarly harvested from the proximal portion of the humerus in 7 of these dogs. Subtrochanteric femoral defects in 11 dogs were randomly assigned to receive cancellous bone graft obtained from the femur (n = 4) or the humerus (n = 7). Subtrochanteric defects in 5 dogs were not grafted. Radiographic assessment of subtrochanteric defects was performed at 4-week intervals, and histologic assessment at 4, 8, 16, and 24 weeks after surgery. Nongrafted donor sites healed by ingrowth of trabecular bone during the first 12 weeks after surgery. By week 24, the lateral cortical wall had reformed, but remodeling was incomplete. Donor sites grafted with cancellous bone healed similarly, but with more rapid healing and more complete remodeling evident by week 24. Although the mean weight of cancellous bone harvested from the proximal portion of the femur (0.82 +/- 0.22 g) was significantly (P less than 0.05) less than that harvested from the proximal portion of the humerus (1.38 +/- 0.29 g), there was no qualitative histologic or radiographic difference in bony healing of grafted defects. We determined that the proximal portion of the femur can be safely used to provide moderate amounts of cancellous bone, and that a second bone graft can be collected from the same subtrochanteric donor site after 12 weeks.     

Micro-computed tomography evaluation and pathological analyses of female rats with collagen-induced arthritis

Imaging techniques have been introduced to assess the efficacy and toxicity of developing pharmaceuticals. The purpose of this study was to perform a comprehensive characterization of collagen-induced arthritis (CIA) in rats using micro-computed tomography (micro-CT) and to compare the results with data from conventional pathological examination. Arthritis was induced by collagen in 24 female Wistar rats. Micro-CT and pathological analyses were performed to assess arthritis progression. Micro-CT analysis showed marked joint destruction occurring in a time-dependent manner following collagen administration. Bone volume was significantly decreased in the tibia at weeks 3 and 4 compared to week 0 (p < 0.05 and p < 0.01, respectively). Additionally, percent bone volume was significantly reduced in the tibia at week 4 compared to week 0 (p < 0.05). In contrast, bone surface/bone volume and trabecular separation were significantly increased in the tibia of the animals at week 4 compared to week 0 (p < 0.05). Severe joint destruction with extensive inflammation, erosion of cartilage and bone, and infiltration of inflammatory cells were observed in the knee joints of the collagen-treated rats. Taken together, micro-CT made it possible to quantify CIA lesions and should be performed with pathological examination in rats.     

Evaluation of biocompatible osteoconductive polymer as an orthopedic implant in dogs.

The biocompatibility and osteoconductive properties of biocompatible osteoconductive polymer (BOP), a synthetic implant, were evaluated. Bilateral oval cortical defects (1 x 2 cm) were made in the lateral subtrochanteric area of the proximal portion of the femur in 16 dogs that later were treated with BOP fiber (n = 16) or autogenous cancellous bone (n = 11), or were not treated (n = 5). The BOP block was attached extraperiosteally to the proximal portion of the humerus in 6 dogs. Radiographic assessment of surgery sites was performed at 4-week intervals, and histologic evaluation was performed at 4, 8, 16, and 24 weeks after surgery. Radiographic signs of bone healing were not observed in defects treated with BOP fiber. Defects treated with cancellous bone or not treated had radiographic signs of progressive bone ingrowth. Radiographic evidence of periosteal new bone formation near control and BOP-treated defects was observed 4 weeks after surgery; increased periosteal reaction was associated with BOP fiber. This new bone had resorbed by week 24, except bone adjacent to BOP fiber, where continued periosteal reaction was apparent. Histologic evidence of bone formation was observed extending to, but not incorporating, BOP fibers. The BOP fibers became surrounded by a fibrous capsule, and fibrovascular connective tissue infiltrated between and into BOP fibers, but minimal bone formation incorporated the BOP material during the follow-up period. During that time, active periosteal new bone formation was evident adjacent to the BOP fibers. Defects treated with cancellous bone or not treated healed by ingrowth of cancellous bone during the first 12 weeks after surgery and by reformation of the lateral cortical wall by week 24. The BOP blocks became surrounded by a fibrous capsule, but connective tissue or bone ingrowth into BOP blocks was not observed. Results indicate that BOP is not osteoconductive within a 6-month time frame when used in subtrochanteric femoral defects or when placed extraperiosteally on the proximal portion of the humerus of clinically normal dogs.     

Immunohistochemical observations on the initial disorders of the epiphyseal growth plate in rats induced by high dose of vitamin A.

The initial disorders of the epiphyseal growth plate cartilage were immunohistochemically examined in the proximal tibia of rats administered a high dose of vitamin A. Male Wistar rats were given 100,000 IU/100 g body weight/day of vitamin A for administration periods of 1 to 5 days (Day 1 to 5) from 4 weeks after birth or were given deionized water and used as control. They were sacrificed after 5-bromo-2'-deoxyuridine (BrdU) injection on Day 1 to Day 5 to remove the tibiae. The tibiae were processed for immunohistochemical examinations using antibodies against type I, II, X collagens and BrdU. BrdU-incorporated chondrocytes and type X collagen-negative area were reduced since Day 2 and type X collagen-positive area was reduced since Day 4. The cartilage matrix partially lost type II collagen and deposited type I collagen in the epiphyseal growth plate near the periosteum on Day 5. These findings suggest that a high dose of vitamin A initially disturbed the differentiation from resting to proliferating chondrocytes, subsequently inhibited the differentiation from proliferating to hypertrophic chondrocytes, caused the chondrocytes to deviate from the process of normal differentiation, and finally resulted in the deformation of the epiphyseal growth plate.     

Use of intramedullary fully-threaded pins in the fixation of feline and canine humeral, femoral and tibial fractures

Intramedullary fully-threaded pins were manufactured from an alloy of titanium, aluminium and vanadium in a fully-threaded style. Pins were produced in various diameters, ranging from 4 mm to 11 mm. Pin lengths varied from 5 cm to 22 cm. The proximal end of the pins was designed to fit into a hexagonal screwdriver, while the distal end was slightly tapered to allow for ease of entry into cancellous bone. Treatment using the fully-threaded intra- medullary pin was carried out in a total of 175 fractures of the humerus, femur, and tibia in 95 cats (bilateral femur in 1 case) and 77 dogs (bilateral femur in 2 cases). Radiographic follow-up for the cases was performed at monthly intervals. Non-union developed in one dog with a femoral fracture in which cerclage wire had also been used. Delayed healing and lameness were observed in two other dogs. Healing with excessive callus formation was observed in 16 dogs. However, there were not any problems noted in these dogs in regards to limb usage. Clinical and radiological results obtained for the remaining cases were found to be very good. Normal, complete fracture healing occurred between four to 14 weeks in dogs, and between four to 12 weeks in cats. Pins were removed upon observation of satisfactory functional and radiographic recovery. Pins could not be removed from 26 cats and 21 dogs as the owners had declined pin removal, or because the owners were lost to follow-up.     

Evaluation of autogeneic and allogeneic cortical chip grafting in a feline tibial nonunion model

A feline tibial nonunion model was used to evaluate the efficacy of fresh autogeneic and deep-frozen allogeneic cortical chip grafts for bone repair. Rigidly fixated tibial ostectomies with a segmental defect length of 1 cm were grafted with 3 mm X 2 mm chips of cortical bone in 12 skeletally mature cats. Six cats were given fresh autogeneic bone (group A) and 6 were given deep-frozen allogeneic bone (group B). Bone healing was evaluated radiographically and histologically over a 12-week period. Consolidating bony callus and palpable stability of the tibia after bone plate removal were evident in all group A cats and 4 of the 6 group B cats by 12 weeks after surgery. Bone remodeling had progressed to intermediate restoration of diaphyseal structure in 2 cats of each group. Fibrous replacement of graft chips, absence of bony callus, and instability of the tibia after bone plate removal were evident in 1 group B cat at 12 weeks. The remaining group B cat was withdrawn from the study at 6 weeks due to loosening of the bone plate and screws.     

Treatment of segmental tibial defects using acute bone shortening followed by gradual lengthening with circular external fixator

The aim of this study was to clinically and radiographically evaluate acute bone shortening followed by gradual lengthening in the treatment of large segmental tibia defects induced in seven clinically normal dogs. A circular external fixator was assembled with one proximal 5/8-circle ring, one middle ring and one distal ring connected with three rods. Thirty per cent of the tibia and fibula were removed in the middle and distal parts of the diaphyses, between the middle and distal rings. Acute bone shortening with compression of proximal and distal segments was performed. A subperiosteal osteotomy was performed between the half-ring and middle ring. Bone distraction started 7 days after surgery; after lengthening, the apparatus was left in place for 14 weeks for consolidation of regenerated bone. The frame was removed at the end of this period, and the dogs observed for four more weeks. Functional results were considered excellent in two, good in three and fair in the other two dogs. Bone regeneration within the distraction gap was obtained 14 weeks after neutral fixation period. We concluded that acute bone shortening followed by gradual lengthening by Ilizarov method can be used to treat extensive tibial defects in dogs, although it presents limb temporary abnormal limb shape and unequal length as early disadvantages.     

APPROACHES FOR ULTRASONOGRAPHIC EVALUATION OF LONG BONES IN THE DOG

The objective of this study was to identify access portals for ultrasonographic evaluation of canine long bones (humerus, radius, ulna, femur, and tibia). A 7.5 MHz linear transducer was used in five medium-sized dogs (range of 25-35 kg body weight). Good approaches could be identified for all long bones. For the proximal humerus, a cranial portal, and the distal humerus, a lateral portal is suggested. For the radius and ulna, a craniolateral approach seemed to be the best. In the hindlimb, a medial approach for the femur and a craniolateral approach for the tibia were the most effective approaches.