Deterioration of wound healing and intense suppression of MMP-9 mRNA expression after short-term administration of different topical glucocorticoids or NSAIDs in an avian model of corneal lesions.

BACKGROUND: Corneal lesions are considered among important ophthalmic conditions in avian patients. Short-term outcome of using anti-inflammatory agents in corneal lesions of birds are not well-described. AIMS: The study evaluates effects of different anti-inflammatory agents on healing of alkali burn-induced corneal lesions in layer hens as an avian model. METHODS: Adult layers were randomly allocated into 7 groups (n=15) as follows: 1. Negative (normal) control (NC), and 2. Positive control (PC) with an experimentally induced-corneal lesion, 3-7. Birds with corneal lesions that were treated with dexamethasone, fluorometholone, prednisolone, ketorolac, or diclofenac eye-drops every 6 hours (QID) for 5 consecutive days. RESULTS: At the end of the experiment, proper healing was observed in PC group based on lesion area, while treated groups showed statistically larger lesion sizes as compared to PC birds (P<0.05). Although no significant difference was observed among groups, birds treated with ketorolac, diclofenac or fluorometholone had higher histopathological scores for most of the assayed parameters than other groups. Levels of tumor necrosis factor-α (TNF-α) and vascular endothelial growth factor (VEGF) in corneal tissue of different groups were statistically the same. The mRNA expression of matrix metalloproteinase-9 (MMP-9) was increased 2.5 folds in PC group as compared to NC birds. However, birds treated with anti-inflammatory agents showed no detectable expression of MMP-9 mRNA. CONCLUSION: Five days of topical administration of non-steroidal anti-inflammatory agents (NSAIDs) or glucocorticoids (GCs) is associated with suppression of MMP-9 mRNA expression in corneal tissue and detrimental effects on wound healing in layers with alkali burn-induced corneal ulcers.

Regional diversity in the murine cortical vascular network is revealed by synchrotron X-ray tomography and is amplified with age.

Cortical bone is permeated by a system of pores, occupied by the blood supply and osteocytes. With ageing, bone mass reduction and disruption of the microstructure are associated with reduced vascular supply. Insight into the regulation of the blood supply to the bone could enhance the understanding of bone strength determinants and fracture healing. Using synchrotron radiation-based computed tomography, the distribution of vascular canals and osteocyte lacunae was assessed in murine cortical bone and the influence of age on these parameters was investigated. The tibiofibular junction from 15-week- and 10-month-old female C57BL/6J mice were imaged post-mortem. Vascular canals and three-dimensional spatial relationships between osteocyte lacunae and bone surfaces were computed for both age groups. At 15 weeks, the posterior region of the tibiofibular junction had a higher vascular canal volume density than the anterior, lateral and medial regions. Intracortical vascular networks in anterior and posterior regions were also different, with connectedness in the posterior higher than the anterior at 15 weeks. By 10 months, cortices were thinner, with cortical area fraction and vascular density reduced, but only in the posterior cortex. This provided the first evidence of age-related effects on murine bone porosity due to the location of the intracortical vasculature. Targeting the vasculature to modulate bone porosity could provide an effective way to treat degenerative bone diseases, such as osteoporosis.

Sexually dimorphic tibia shape is linked to natural osteoarthritis in STR/Ort mice.

OBJECTIVES: Human osteoarthritis (OA) is detected only at late stages. Male STR/Ort mice develop knee OA spontaneously with known longitudinal trajectory, offering scope to identify OA predisposing factors. We exploit the lack of overt OA in female STR/Ort and in both sexes of parental, control CBA mice to explore whether early divergence in tibial bone mass or shape are linked to emergent OA. METHOD: We undertook detailed micro-CT comparisons of trabecular and cortical bone, multiple structural/architectural parameters and finite element modelling (FEM) of the tibia from male and female STR/Ort and CBA mice at 8-10 (pre-OA), 18-20 (OA onset) and 40 + weeks (advanced OA) of age. RESULTS: We found higher trabecular bone mass in female STR/Ort than in either OA-prone male STR/Ort or non-prone CBA mice. Cortical bone, as expected, showed greater cross-sectional area in male than female CBA, which surprisingly was reversed in STR/Ort mice. STR/Ort also exhibited higher cortical bone mass than CBA mice. Our analyses revealed similar tibial ellipticity, yet greater predicted resistance to torsion in male than female CBA mice. In contrast, male STR/Ort exhibited greater ellipticity than both female STR/Ort and CBA mice at specific cortical sites. Longitudinal analysis revealed greater tibia curvature and shape deviations in male STR/Ort mice that coincided with onset and were more pronounced in late OA. CONCLUSION: Generalised higher bone mass in STR/Ort mice is more marked in non OA-prone females, but pre-OA divergence in bone shape is restricted to male STR/Ort mice in which OA develops spontaneously.

Crack driving force in twisted plywood structures.

Twisted plywood architectures can be observed in many biological materials with high fracture toughness, such as in arthropod cuticles or in lamellar bone. Main purpose of this paper is to analyze the influence of the progressive rotation of the fiber direction on the spatial variation of the crack driving force and, thus, on the fracture toughness of plywood-like structures. The theory of fiber composites is used to describe the stiffness matrix of a twisted plywood structure in a specimen-fixed coordinate system. The driving force acting on a crack propagating orthogonally to the fiber-rotation plane is studied by methods of computational mechanics, coupled with the concept of configurational forces. The analysis unfolds a spatial variation of the crack driving force with minima that are beneficial for the fracture toughness of the material. It is shown that the estimation of the crack driving force can be simplified by replacing the complicated anisotropic twisted plywood structure by an isotropic material with appropriate periodic variations of Young's modulus, which can be constructed based either on the local stiffness or local strain energy density variations. As practical example, the concepts are discussed for a specimen with a stiffness anisotropy similar to lamellar bone. STATEMENT OF SIGNIFICANCE: Twisted plywood-like structures exist in many natural fiber composites, such as bone or insect carapaces, and are known to be very fracture resistant. The crack driving force in such materials is analyzed quantitatively for the first time, using the concept of configurational forces. This tool, well established in the mechanics of materials, is introduced to the modeling of biological material systems with inhomogeneous and anisotropic material behavior. Based on this analysis, it is shown that the system can be approximated by an appropriately chosen inhomogeneous but isotropic material for the calculation of the crack driving force. The spatial variation of the crack driving force and, especially, its local minima are essential to describe the fracture properties of twisted plywood structures.

Does sequence polymorphism of FLC paralogues underlie flowering time QTL in Brassica oleracea?

Previous locations of flowering time (FT) QTL in several Brassica species, coupled with Arabidopsis synteny, suggest that orthologues of the genes FLC, FY or CONSTANS might be the candidates. We focused on FLC, and cloned paralogous copies in Brassica oleracea, obtained their genomic DNA sequences, and confirmed their locations relative to those of known FT-QTL by genetical mapping. They varied in total length mainly due to the variable size of the first and last introns. A high level of identity was observed among Brassica FLC genes at the amino acid level but non-synonymous differences were present. Comparative analysis of the promoter and intragenic regions of BoFLC paralogues with Arabidopsis FLC revealed extensive differences in overall structure and organisation but showed high conservation within those segments known to be essential in regulating FLC expression. Four B. oleracea FLC copies (BoFLC1, BoFLC3, BoFLC4 and BoFLC5) were located to their respective linkage groups based on allelic sequence variation in lines from a doubled haploid population. All except BoFLC4 were within the confidence intervals of known FT-QTL. Sequence data indicated that relevant non-synonymous polymorphisms were present between parents A12DHd and GDDH33 for BoFLC genes. However, BoFLC alleles segregated independently of FT in backcrosses while the study provided evidence that BoFLC4 and BoFLC5 contain premature stop codons and so could not contribute to flowering time variation. Therefore, there is strong evidence against any of the 4 BoFLC being FT-QTL candidates in this population.