Michael W. Orth

The extent of matrix damage and chondrocyte death in mechanically traumatized articular cartilage explants depends on rate of loading

Mechanical loads can lead to matrix damage and chondrocyte death in articular cartilage. This damage has been implicated in the pathogenesis of secondary osteoarthritis. Studies on cartilage explants with the attachment of underlying bone at high rates of loading have documented cell death adjacent to surface lesions. On the other hand, studies involving explants removed from bone at low rates of loading suggest no clear spatial association between cell death and matrix damage. The current study hypothesized that the observed differences in the distribution of cell death in these studies are attributed to the rate of loading. Ninety bovine cartilage explants were cultured for two days. Sixty explants were loaded in unconfined compression to 40 MPa in either a fast rate of loading experiment (∼900 MPa/s) or a low rate of loading experiment (40 MPa/s). The remaining 30 explants served as a control population. All explants were cultured for four days after loading. Matrix damage was assessed by measuring the total length and average depth of surface lesions and the release of glycosaminoglycans to the culture media. Explants were sectioned and stained with calcein and ethidium bromide homodimer to document the number of live and dead cells. Greater matrix damage was documented in explants subjected to a high rate of loading, compared to explants exposed to a low rate of loading. The high rate of loading experiments resulted in cell death adjacent to fissures, whereas more dead cells were observed in the low rate of loading experiments and a more diffuse distribution of dead cells was observed away from the fissures. In conclusion, this study indicated that the rate of loading can significantly affect the degree of matrix damage, the distribution of dead cells, and the amount of cell death in unconfined compression experiments on explants of articular cartilage.

Effect of rearing environment on bone growth of pullets

Alternative housing systems for laying hens provide mechanical loading and help reduce bone loss. Moreover, achieving greater peak bone mass during pullet phase can be crucial to prevent fractures in the production period. The aim of this study was to determine the housing system effects on bone quality of pullets. Tibiae and humeri of White Leghorn pullets reared in conventional cages (CCs) and a cage-free aviary (AV) system were studied. At 16 wk, 120 birds at random from each housing system were euthanized. Right and left tibiae and humeri were collected and further analyzed. Cortical bone density and thickness were measured using computed tomography. Periosteal and endosteal dimensions were measured at the fracture site during mechanical testing. At 4, 8, 12, and 16 wk, serum concentrations of osteocalcin and hydroxylysyl pyridinoline were analyzed as markers of bone formation and resorption. Cortical bone density was higher (P < 0.05) in humeri of AV pullets, and tibiae were denser (P < 0.05) for AV pullets in the distal section of the bone compared to CC pullets. Ash content was higher (P < 0.05) in AV humeri with no difference in tibiae ash content. Tibiae and humeri of AV pullets had a thicker cortex than the CC pullets (P < 0.05). Additionally, the tibiae and humeri of AV pullets had greater (P < 0.05) second moment of areas than the CC pullets. While some bone material properties between groups were different (P < 0.05), the differences were so small (< 7%) that they likely have no clinical significance. Serum osteocalcin concentrations were not different between the treatments, but hydroxylsyl pyridinoline concentrations were higher in CC pullets at 12 wk compared to the AV pullets and the effect reversed at 16 wk (P < 0.05). These findings indicate that tibiae and humeri respond differently to load bearing activities during growth. The improved load bearing capability and stiffness in bones of AV pullets were related to increased cross-sectional geometry.

Housing conditions alter properties of the tibia and humerus during the laying phase in Lohmann white Leghorn hens

Osteoporosis in caged hens is one driving factor for the United States egg industry to explore options regarding alternative housing systems for laying hens. The aim of our research was to study the influence of housing systems on tibiae and humeri of 77-week-old Lohmann White hens. Pullets raised in an aviary system were either continued in aviary hen systems (AV) or conventional cages (AC) whereas pullets reared in conventional cages continued in conventional hen cages (CC) or enriched colony cages (EN) at 19 weeks. From each group, 120 hens were randomly euthanized and right and left tibae and humeri were excised for structural and mechanical analysis. Volumetric density of the cortical bone was measured using quantitative computed tomography (QCT). Aviary (AV) hens had greater cortical thickness and density but similar outer dimensions to AC hens (P < 0.05). Hens in EN system had humeri with similar cortical thickness and density but wider outer dimensions than the humeri of CC hens (P < 0.05). Cortical geometry of the tibiae was the same for the EN and CC hens, whereas EN hens had denser tibial cortex than CC hens (P < 0.05). Geometrical changes in the humeri suggest that hens in the AV system were better able to protect their structure from endosteal resorption during the laying phase. Humeri of AV and EN hens had increased second moment of area compared to the AC and CC hens; however, the changes were not observed in tibiae. Mechanical property differences were observed, with bones of AV hens having greater failure moment and stiffness than AC hens and the same difference was observed between the EN and CC hens, (P < 0.05). These findings indicate that movement limitation causes loss of bone mass and density whereas provision of moderate movement increases certain bone quality parameters during adulthood in laying hens.

Evaluation of inflammatory responses induced via intra-articular injection of interleukin-1 in horses receiving a dietary nutraceutical and assessment of the clinical effects of long-term nutraceutical administration.

OBJECTIVE To evaluate inflammatory responses induced via intra-articular recombinant human interleukin (IL)-1beta treatment in horses receiving a dietary nutraceutical (DN; composed of mussel, shark cartilage, abalone, and Biota orientalis lipid extract) and assess the clinical effects of long-term DN administration. ANIMALS 22 healthy horses. PROCEDURES 12 horses were fed 0, 15, 45, or 75 mg of DN (3 horses/treatment) daily for 84 days. General health and clinicopathologic variables were monitored at intervals. Ten other horses received 0 or 15 g of DN/d (5 horses/treatment) for 29 days (beginning day -14). One intercarpal joint in each horse was injected twice with IL-1beta (10 and 100 ng on days 0 and 1, respectively), and the contralateral joint was similarly injected with saline (0.9% NaCl) solution. Synovial fluid prostaglandin E(2) (PGE(2)), sulfated glycosaminoglycan (GAG), nitric oxide (NO), and protein concentrations and leukocyte counts were analyzed before and at intervals after injections. RESULTS Administration of the DN (up to 75 g/d) to horses for 84 days did not induce any adverse effects. In the other experiment, synovial fluid PGE(2), GAG, and protein concentrations and leukocyte count increased after intra-articular injections of IL-1beta (compared with effects of saline solution injections) in horses that received no DN; NO concentration was not affected. In horses that were fed the DN, intra-articular IL-1beta injections did not induce significant increases in synovial fluid PGE(2) and GAG concentrations. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that administration of the DN may be useful in preventing inflammation associated with arthritis and degenerative joint disease in horses.

Equine osteoarthritis: a brief review of the disease and its causes.

Degenerative joint diseases, such as osteoarthritis, adversely impact the health of the equine athlete as well as the economics of the equine industry. Our understanding of the aetiology of osteoarthritis, although not nearly exhaustive, has increased substantially in recent years. Molecules, including cytokines, inflammatory mediators, and metalloproteinases, have been identified and associated with the progression of joint disease. Several factors, including trauma to the joint, immobilization, conformation, shoeing, and ageing, have been linked with osteoarthritis. Our continued efforts into elucidating critical biological mediators and risk factors, coupled with better chondroprotective therapies and diagnostic tools, should facilitate our ability to maintain the skeletal health of the equine athlete

Growth and maturational changes in dense fibrous connective tissue following 14 days of rhGH supplementation in the dwarf rat

The purpose of this study was to investigate the impact of recombinant human growth hormone (rhGH) on patella tendon (PT), medial collateral ligament (MCL), and lateral collateral ligament (LCL) on collagen growth and maturational changes in dwarf GH-deficient rats. Twenty male Lewis mutant dwarf rats, 37 days of age, were randomly assigned to Dwarf + rhGH (n = 10) and Dwarf + vehicle (n = 10) groups. The GH group received 1.25 mg rhGH/kg body wt twice daily for 14 days. rhGH administration stimulated dense fibrous connective tissue growth, as demonstrated by significant increases in hydroxyproline specific activity and significant decreases in the non-reducible hydroxylysylpyridinoline (HP) collagen cross-link contents. The increase in the accumulation of newly accreted collagen was 114, 67, and 117% for PT, MCL, and LCL, respectively, in 72 h. These findings suggest that a short course rhGH treatment can affect the rate of new collagen production. However, the maturation of the tendon and ligament tissues decreased 18-25% during the rapid accumulation of de novo collagen. We conclude that acute rhGH administration in a dwarf rat can up-regulate new collagen accretion in dense fibrous connective tissues, while causing a reduction in collagen maturation.

Nonreducible crosslink formation in tibial dyschondroplastic growth plate cartilage from broiler chicks fed homocysteine

In the study of tibial dyschondroplasia, scientists have for a long time thought that an altered extracellular matrix might be involved in the etiology of the disease. The results presented in this paper show that the collagen content was increased in the dyschondroplastic cartilage when compared to normal growth plate and day-old hypertrophic cartilage. Furthermore, nonreducible crosslinks were found only in dyschondroplastic cartilage, with the greatest amounts occurring in the distal region of the lesion, approximately 10-fold higher than that found in the dyschondroplastic growth plate. Thus, intermolecular collagen bonding is altered in the extracellular matrix of dyschondroplastic cartilage. Possible models for the etiology of the disease are discussed.

Increased plasma silicon concentrations and altered bone resorption in response to sodium zeolite a supplementation in yearling horses

Abstract This study examined the effect of supplementation of a bioavailable source of silicon (sodium zeolite A) on altering systemic markers of bone metabolism in horses. Twenty yearlings (ten Quarter Horses and ten Arabians) were randomly grouped as silicon (Si) supplemented (S; n=10), in which yearlings consumed 2% of the total diet as a Si-containing supplement, and a second non-supplemented control group (C; n=10). Blood samples were taken on days 0, 15, 30 and 45. Both plasma and serum were collected; the plasma was analyzed for Si concentrations and serum was analyzed for osteocalcin (OC), carboxy-terminal pyridinoline cross-linked telopeptide region of type I collagen (ICTP), and pyridinoline and deoxypyridinoline crosslinks (PYD). Supplemented yearlings had higher plasma Si concentrations than C yearlings by day 15, and remained higher than C yearlings on days 30 and 45 (P .05); however, ICTP concentrations were lower in S yearlings on day 45 when compared to C yearlings (P = .04). Results indicate that sodium zeolite A supplementation (consumed at 2% of the total diet) increases plasma Si concentrations. Furthermore, results indicate that Si-supplemented yearlings may have decreased bone resorption, which may provide for greater net bone formations, as OC concentrations were not different between groups. Unfortunately, systemic markers give no indication as to the quality of the bone that may be formed, and further research in the area of Si supplementation, bone metabolism and bone strength is required to establish conclusive evidence as to the benefits of supplemental Si to the skeletal system.

Non-reducible Collagen Cross-linking in Cartilage from Broiler Chickens with Tibial Dyschondroplasia

Recent work has shown that hydroxylysylpyridinoline (HP), a non-reducible cross-link that stabilizes the collagen fibril network, is significantly greater (over 10-fold) in dyschondroplastic cartilage than in normal growth-plate cartilage in the tibiotarsi of chickens with homocysteine-induced tibial dyschondroplasia (TD). In the present study, broiler chicks with a genetic disposition to TD, as well as normal broiler chicks on a copper-deficient diet alone or supplemented with copper and thiram, were raised for 3 to 4 weeks (Expts. 1 and 2). Their dyschondroplastic cartilage from the proximal tibiotarsus was collected and analyzed for HP as well as lysylpyridinoline (LP) cross-links. Normal growth plate cartilage was obtained from chicks on the copper-deficient diet supplemented with copper. In a third experiment, another set of broiler chicks was raised on a corn/soybean meal-based diet with or without homocysteine, and their articular and sternal cartilage were isolated for cross-link analysis. In the first two experiments, dyschondroplastic cartilage from all birds with induced TD had higher HP and LP concentrations than growth-plate cartilage from birds without TD, although the ratios of HP to LP varied. In the third experiment, the sternal and articular cartilage from birds with homocysteine-induced TD appeared normal, having similar HP concentrations in the same types of cartilage in birds without TD.

The presence of lysylpyridinoline in the hypertrophic cartilage of newly hatched chicks

The presence of lysylpyridinoline (LP) as a nonreducible cross-link in appreciable quantities has primarily been limited to the mineralized tissues, bone and dentin. However, the results reported here show that LP is not only present in the hypertrophic cartilage of the tibiotarsus isolated from newly hatched broiler chicks, but it is approx. 4-fold as concentrated as hydroxylysylpyridinoline (HP). Bone and articular cartilage surrounding the hypertrophic cartilage do not contain measurable quantities of LP. Purified LP has a fluorescent scan similar to purified HP and literature values, confirming that we indeed were measuring LP. Also, the cartilage lesion produced by immature chondrocytes from birds with tibial dyschondroplasia had LP but the HP:LP ratio was > 1. Thus, the low HP:LP ratio could be a marker for hypertrophic cartilage in avians.