The contribution of collagen fibers to the mechanical compressive properties of the temporomandibular joint disc.

OBJECTIVE: The Temporomandibular Joint (TMJ) disc is a fibrocartilaginous structure located between the mandibular condyle and the temporal bone, facilitating smooth movements of the jaw. The load-bearing properties of its anisotropic collagenous network have been well characterized under tensile loading conditions. However, recently it has also been speculated that the collagen fibers may contribute dominantly in reinforcing the disc under compression. Therefore, in this study, the structural-functional role of collagen fibers in mechanical compressive properties of TMJ disc was investigated. DESIGN: Intact porcine TMJ discs were enzymatically digested with collagenase to disrupt the collagenous network of the cartilage. The digested and non-digested articular discs were analyzed mechanically, biochemically and histologically in five various regions. These tests included: (1) cyclic compression tests, (2) biochemical quantification of collagen and glycosaminoglycan (GAG) content and (3) visualization of collagen fibers' alignment by polarized light microscopy (PLM). RESULTS: The instantaneous compressive moduli of the articular discs were reduced by as much as 50-90% depending on the region after the collagenase treatment. The energy dissipation properties of the digested discs showed a similar tendency. Biochemical analysis of the digested samples demonstrated an average of 14% and 35% loss in collagen and GAG, respectively. Despite the low reduction of collagen content the PLM images showed considerable perturbation of the collagenous network of the TMJ disc. CONCLUSIONS: The results indicated that even mild disruption of collagen fibers can lead to substantial mechanical softening of TMJ disc undermining its reinforcement and mechanical stability under compression.

Comparative effects of zinc-nano complexes, zinc-sulphate and zinc-methionine on performance in broiler chickens.

Micronutrients, especially zinc, have an important role in normal metabolism and growth of broilers. Using novel technologies helps to synthesise novel zinc complexes to deliver this micronutrient more efficiently. In the present study, the effects of different zinc complexes and nano complexes on broiler performance were compared. Broilers in 6 groups were given basal diet (without zinc) and basal diet supplemented with zinc-sulphate, zinc-methionine, zinc-nano-sulphate, zinc-nano-methionine and zinc-nano-max (that was synthesised based on nanochelating technology) at a concentration of 80 mg/kg of diet. At 1-42 d of age, dietary zinc-nano-sulphate supplementation decreased weight gain and feed intake. However, feed conversion ratio was not influenced by treatments. Carcass yield (%) of birds in the zinc-nano-sulphate and control groups were dramatically reduced at 42 d of age and abdominal fat (%) increased in these groups. Relative to the control group, the antibody titre, spleen and bursa of Fabricius (%) were significantly higher in groups supplemented with zinc. Heterophil (%) was also significantly higher in the zinc-nano-methionine group in blood on d 42 compared to the control, zinc-sulphate and zinc-nano-sulphate. Compared to the controls, the mean malondialdehyde content in thigh tissue was significantly reduced in groups supplemented with zinc at the time 0, 50, 100 and 150 min after oxidation. Tibia zinc concentration in nanoparticle zinc samples was significantly higher relative to the control and zinc-sulphate groups. Taken together, our data indicate that delivery of zinc in the structure of zinc-nano-methionine and zinc-nano-max at concentrations of 80 mg/kg of diet improves growth performance. However, dietary zinc-nano-sulphate decreased growth performance in broilers.

A comparative analysis of the collagen architecture in the carotid artery: second harmonic generation versus diffusion tensor imaging.

Collagen is the main load-bearing component of the artery. The 3D arrangement of the collagen fibers is crucial to understand the mechanical behavior of such tissues. We compared collagen fiber alignment obtained by second harmonic generation (SHG) microscopy with the alignment obtained by diffusion tensor imaging (DTI) throughout the wall of a porcine carotid artery to check the feasibility of using DTI as a fast and non-destructive method instead of SHG. The middle part of the artery was cut into two segments: one for DTI and one for the SHG measurements. The tissue for SHG measurements was cut into 30µm tangential sections. After scanning all sections, they were registered together and the fiber orientation was quantified by an in-house algorithm. The tissue for DTI measurement was embedded in type VII agarose and scanned with an MRI-scanner. Fiber tractography was performed on the DTI images. Both methods showed a layered structure of the wall. The fibers were mainly oriented circumferentially in the outer adventitia and media. DTI revealed the predominant layers of the arterial wall. This study showed the feasibility of using DTI for evaluating the collagen orientation in native artery as a fast and non-destructive method.

Evaluation of adiponectin gene expression in the abdominal adipose tissue of broiler chickens: feed restriction, dietary energy, and protein influences adiponectin messenger ribonucleic acid expression.

We studied the effects of feed restriction and different energy and protein contents of the diet on BW, abdominal adipose tissue percentage, and adiponectin gene expression in abdominal adipose tissue in broiler chickens. Two experiments were conducted to determine whether feed restriction and dietary energy and protein levels alter adiponectin mRNA abundance in broiler chicks. Body weight and abdominal adipose tissue percentage were recorded and abdominal adipose tissue samples were collected at sampling days. Adiponectin mRNA expression in abdominal adipose tissue was quantitated using real-time quantitative PCR. We found that BW, abdominal adipose tissue percentage, and adiponectin gene expression were decreased in restricted chicks compared with those fed ad libitum at 32 d of age, but feed restriction had no effect on abdominal adipose tissue percentage and adiponectin gene expression at 49 d of age (experiment 1). Body weight was increased significantly in broilers fed on low-energy diets compared with those fed high-energy diets in experiment 1. Also, increasing levels of dietary protein increased BW in broiler chicks in experiment 2. A positive response in adiponectin gene expression and abdominal adipose tissue percentage was achieved by decreasing protein level in the diet of chicks at 32 d of age, but dietary protein had no effect on adiponectin gene expression at 49 d of age (experiment 1). Also, in experiment 2, abdominal adipose tissue percentage increased as dietary protein with fixed level of energy decreased in broiler chickens at 42 and 56 d of age. In experiment 1, increasing dietary energy levels increased adiponectin gene expression and abdominal adipose tissue percentage in chicks at 32 d of age. Also, we showed that dietary energy had an effect on abdominal adipose tissue percentage at 49 d of age. In experiment 2, a high-energy diet with a fixed level of protein increased abdominal adipose tissue percentage in chicks at 42 and 56 d of age. Dietary energy and protein levels had no significant effect on adiponectin gene expression in abdominal adipose tissue in broilers.

Modulation of collagen fiber orientation by strain-controlled enzymatic degradation.

Collagen fiber anisotropy has a significant influence on the function and mechanical properties of cardiovascular tissues. We investigated if strain-dependent collagen degradation can explain collagen orientation in response to uniaxial and biaxial mechanical loads. First, decellularized pericardial samples were stretched to a fixed uniaxial strain and after adding a collagen degrading enzyme (collagenase), force relaxation was measured to calculate the degradation rate. This data was used to identify the strain-dependent degradation rate. A minimum was observed in the degradation rate curve. It was then demonstrated, for the first time, that biaxial strain in combination with collagenase alters the collagen fiber alignment from an initially isotropic distribution to an anisotropic distribution with a mean alignment corresponding with the strain at the minimum degradation rate, which may be in between the principal strain directions. When both strains were smaller than the minimum degradation point, fibers tended to align in the direction of the larger strain and when both strains were larger than the minimum degradation, fibers mainly aligned in the direction of the smaller strain. However, when one strain was larger and one was smaller than the minimum degradation point, the observed fiber alignment was in between the principal strain directions. In the absence of collagenase, uniaxial and biaxial strains only had a slight effect on the collagen (re)orientation of the decellularized samples. STATEMENT OF SIGNIFICANCE: Collagen fiber orientation is a significant determinant of the mechanical properties of native tissues. To mimic the native-like collagen alignment in vitro, we need to understand the underlying mechanisms that direct this alignment. In the current study, we aimed to control collagen fiber orientation by applying biaxial strains in the presence of collagenase. We hypothesized that strain-dependent collagen degradation can describe specific collagen orientation when biaxial mechanical strains are applied. Based on this hypothesis, collagen fibers align in the direction where the degradation is minimal. Pericardial tissues, as isotropic collagen matrices, were decellularized and subjected to a fixed uniaxial strain. Then, collagenase was added to initiate the collagen degradation and the relaxation of force was measured to indicate the degradation rate. The V-shaped relationship between degradation rate and strain was obtained to identify the minimum degradation rate point. It was then demonstrated, for the first time, that biaxial strain in combination with collagenase alters the collagen fiber alignment from almost isotropic to a direction corresponding with the strain at the minimum degradation rate.

Analysis and frequency of bovine lymphocyte antigen (BoLA-DRB3) alleles in Iranian Holstein cattle.

The bovine lymphocyte antigen (BoLA-DRB3) gene encodes cell surface glycoproteins that initiate immune response by presenting processed antigenic peptides to CD4 T helper cells. DRB3 is the most polymorphic bovine MHC class II gene which encodes the peptide-binding groove. DRB3 gene has been extensively evaluated as a candidate marker for association with various bovine diseases and immunological traits. This study describes genetic variability in the BoLA-DRB3 in Iranian Holstein cattle. This is the first study of the DNA polymorphism of the BoLA-DRB3 gene in Iranian Holstein cattle. Hemi-nested PCR-RFLP method is used for identification the frequency of BoLA-DRB3 alleles. The BoLA-DRB3 locus is highly polymorphic in the studied herd (26 alleles). Almost 67% of the alleles were accounted for four alleles (BoLA-DRB3.2*8, *24, *11 and *16) in Iranian Holstein cattle. The DRB3.2*8 allele frequency (26.6%) was higher than the others. The frequencies of the DRB3.2*54, *37, *36, *28, *25, *14, *13, *10, *1 alleles were lower than 1%. Significant distinctions have been found between Iranian Holstein cattle and other cattle breeds studied. In Iranian Holstein cattle the alleles (BoLA-DRB3.2*22, *2 and *16) associated with a lower risk of cystic ovarian disease in Holstein cattle are found. The alleles associated with the resistance to mastitis and to bovine leukemia virus infection BoLA-DRB3.2*11 and *23 are detected with the frequencies 10.4% and 4.4%, respectively. Thus in the Iranian Holstein cows studied are found alleles which are associated with resistance to various diseases. The method of DNA-typing of animals can be used in agricultural practice for BoLA-DRB3 allele genotyping of cattle in order to reduce spreading of alleles providing susceptibility to mastitis or leukemia in cattle herds.

Effect of cyanocobalamin (vitamin B12) in the induction and expression of morphine tolerance and dependence in mice.

The antinociceptive effect of cyanocobalamin (Vit B12) has been reported in animal models and human studies. Our previous study showed the effect of Vit B12 on morphine tolerance. The dependence and tolerance were induced in male mice using subcutaneous morphine injections, 3 times a day (50, 50 and 75 mg/kg/day) for 3 days. Mice also received Vit B12 (100, 250 and 500 µg/kg), clonidine, memantine and saline intraperitoneally before morphine administration. On fourth day mice received only 7 mg /kg morphine just before tail-flick test. To determine the expression of morphine dependence and tolerance, all compounds were injected once intraperitoneally on the day of experiment. The tolerance was evaluated by the tail-flick test. The effect of Vit B12 and other agents on dependence were evaluated by counting the number of jumps (induced by naloxone 5 mg/kg). Co-administration of Vit B12 (100-500 µg/kg) and morphine in 3 days reduced the development of tolerance to morphine analgesic effect (8.2±0.5 and 7.83±0.5 s. vs. normal saline, 3.57±0.3 s). Repeated administration of Vit B12, also, diminished the reduced naloxane withdrawal signs of naloxone withdrawal test (100-500 µg/kg: 5±1.9 and 1.2±0.8 jumps vs. normal saline 72.6±12.2). However, Vit B12 had no effect on the expression of morphine tolerance and physical dependence. It is concluded that co-administration of Vit B12 and morphine could reduce tolerance to analgesic effect of morphine chronic administration and also reduce its withdrawal symptoms.