In this article the mechanical properties of contracted collagen gels were investigated thoroughly by means of uniaxial tensile test. Large type I collagen-Dulbecco's Modified Eagle Medium (DMEM) gels (each was 26 ml in volume, 1.67 mg/ml collagen concentration), each populated with about 2.5 x 106 human fibroblasts, were made in 100 mm diameter plastic dishes precoated with albumin for floating the gels in DMEM. Such identically treated gels were divided into three groups for the mechanical measurements at different culture periods (2, 4, and 10 weeks). Rapid contraction occurred within the first 3 days and then the contraction went slowly in the rest period until it reached about 13% of its original size. The stress-strain curve of the contracted collagen gels demonstrated an exponential behavior at low stress region, followed by linear region, a point of yielding, and finally an ultimate stress point at which the maximum stress was reached. The mechanical strength increased in the first few weeks and then decreased as the culture went on. It is obvious that the collagen fibrils formed and were forced to orientate to the tensile direction after the test. The stress relaxation and cyclic creep phenomena were observed. Based on the morphological analysis of transmission electron microscopy (TEM) of the gels, a nonlinear visco-elastic-plastic constitutive formula was proposed, which was able to reproduce the rheological phenomena of the gels. This experiment shows that the human fibroblasts significantly contracted collagen gels so as to achieve certain mechanical strength, which makes it possible to be a scaffold for tissue engineering. However, a further method to reinforce the mechanical strength by several folds must be considered. Meanwhile, the rheological phenomena should be taken into account in the fabrication and application of the structure.
Immunohistochemical distribution and cellular localization of neurotrophins was investigated in adult monkey brains using antisera against nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and neurotrophin-4 (NT-4). Western blot analysis showed that each antibody specifically recognized appropriate bands of approximately 14.7 kDa, 14.2 kDa, 13.6 kDa, and 14.5 kDa, for NGF, BDNF, NT-3, and NT-4, respectively. These positions coincided with the molecular masses of the neurotrophins studied. Furthermore, sections exposed to primary antiserum preadsorbed with full-length NGF, BDNF, NT-3, and NT-4 exhibited no detectable immunoreactivity, demonstrating specificities of the antibodies against the tissues prepared from rhesus monkeys. The study provided a systematic report on the distribution of NGF, BDNF, NT-3, and NT-4 in the monkey brain. Varying intensity of immunostaining was observed in the somata and processes of a wide variety of neurons and glial cells in the cerebrum, cerebellum, hippocampus, and other regions of the brain. Neurons in some regions such as the cerebral cortex and the hippocampus, which stained for neurotrophins, also expressed neurotrophic factor mRNA. In some other brain regions, there was discrepancy of protein distribution and mRNA expression reported previously, indicating a retrograde or anterograde action mode of neurotrophins. Results of this study provide a morphological basis for the elucidation of the roles of NGF, BDNF, NT-3, and NT-4 in adult primate brains.
The role of sodium channel voltage-gated beta 2 (SCN2B) in brain aging is largely unknown. The present study was therefore designed to determine the role of SCN2B in brain aging by using the senescence-accelerated mice prone 8 (SAMP8), a brain senescence-accelerated animal model, together with the SCN2B transgenic mice. The results showed that SAMP8 exhibited impaired learning and memory functions, assessed by the Morris water maze test, as early as 8 months of age. The messenger RNA (mRNA) and protein expressions of SCN2B were also upregulated in the prefrontal cortex at this age. Treatment with traditional Chinese anti-aging medicine Xueshuangtong (Panax notoginseng saponins, PNS) significantly reversed the SCN2B expressions in the prefrontal cortex, resulting in improved learning and memory. Moreover, SCN2B knockdown transgenic mice were generated and bred to determine the roles of SCN2B in brain senescence. A reduction in the SCN2B level by 60.68% resulted in improvement in the hippocampus-dependent spatial recognition memory and long-term potential (LTP) slope of field excitatory postsynaptic potential (fEPSP), followed by an upregulation of COX5A mRNA levels and downregulation of fibroblast growth factor-2 (FGF-2) mRNA expression. Together, the present findings indicated that SCN2B could play an important role in the aging-related cognitive deterioration, which is associated with the regulations of COX5A and FGF-2. These findings could provide the potential strategy of candidate target to develop antisenescence drugs for the treatment of brain aging.
In this paper the mechanical properties of contracted collagen gels populated with rat fibroblasts or cardiomyocytes were investigated by means of uniaxial tensile testing. Rat type I collagen-Dulbecco's modified Eagle's medium (DMEM) gels (each 2 ml in volume, 0.5 mg/ml collagen concentration) populated with different numbers of rat fibroblasts or cardiomyocytes were made in 31 x 17-mm wells cut in silicone rubber located in a 100-mm diameter plastic dish. Identically treated gels were incubated for 4 days floating in DMEM and then were subjected to uniaxial tensile testing. Rapid contraction occurred within the first 3 days for both the fibroblast and cardiomyocyte gels, but the cardiomyocyte gels consistently contracted to smaller sizes than the fibroblast gels for each number of cells used. The tension-strain curve of the contracted collagen gels demonstrated exponential behavior in the low stress region, followed by a linear section, and finally a maximum tension point, giving the ultimate strength of the gel tested. The cardiomyocyte gels had higher tension-strain curves than the fibroblast gels for each number of cells used. The tension relaxation and cyclic creep phenomena were observed in both kinds of gels, and these phenomena coincide with prior observations in collagen gels contracted by human fibroblasts. This experiment shows that type I collagen gels can be significantly contracted by rat fibroblasts or cardiomyocytes so as to achieve a certain mechanical strength. The contracted collagen structures made in these experiments have potential for developing tissue-engineered structures for cardiac muscle studies.
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