| Summary: | 博士 === 南台科技大學 === 電機工程系 === 102 === According to the latest annual statistics report of the Ministry of Interior, as of 2013, there is an estimated 382,000 people with physical disability. This figure belongs to the largest percentage (34%) of the disabled population. Among them, about 40,000 people are patients with spinal cord injury (SCI), and annually, a continuous increase of around a thousand people is expected. In general, regardless of the field of experimental medical research, most studies are done through experiments on animals. The analysis of the motor behavior among animals is also often used in comparing experimental studies. Studies related to spinal injuries mostly use male rats of the Sprague-Dawley strains (SD) as the object of animal experiments. The Basso-Beattie-Bresnahan (BBB) open field locomotor scale test is used as the criterion to evaluate hind limb motor function. In investigating the relationship between cellular and molecular motor functions, this study firstly attempted to assess and compare the protective effect on hind limb functional recovery after human umbilical cord blood cells (HUCBC) or DMEM medium (without cell culture, as a vehicle) which is given through purified hematopoietic stem cells (CD34+), secreting cells or substances called conditioned medium (CM) cultured from human umbilical cord blood-derived CD34+ cells, combined CM and 17β-estradiol medium were given intravenously immediately after spinal cord injury. In this study, the following conditions for recovery after spinal cord injury were designed: (1) the increase in number and activity of apoptotic protease (caspase-3) of DNA terminal deoxynucleotidyl transferase-mediated dUTP DNA labeled cells (TUNEL - positive cell is evidence) is used as evidence in determining apoptosis; (2) the increase in performance of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α as support) and the activity of myeloperoxidase (MPO) are evidences of activated inflammatory response; (3) the increase in nitric oxide (NO), 2,3-dihydroxybenzoic acid, and malondialdehyde (MDA as evidence) and the reduction of superoxide dismutase (SOD), glutathione peroxidase (GPx), and glutathione reductase (GR) are evidences of increased oxidative stress; (4) the amount of glial fibrillary acidic protein-positive cells (GFAP-positive cells) shows evidence of increased astrogliosis. The entire study found out that rats given conditioned medium, 17β-estradiol, and combined CM and 17β-estradiol medium immediately after SCI can effectively improve their situation on the 7th day of injury. Also, injured spinal cord is neuroprotected, and this effectively improves hind limb motor function. Subsequently, in treating SCI, combined CM and 17β-estradiol medium is better than a single CM or 17β-estradiol. These results illustrate that 17β-estradiol can strengthen the cultivation of CD34+ stem cell-conditioned medium derived from human umbilical cord blood cells, increasing the therapeutic effect on injured spinal cord of experimental rats.
In coordination with the above-mentioned work, this study also attempted to establish a set of computer-assisted quantitative assessment system for animal behavior analysis on the basis of imaging technology in order to test the hind limb behavior of observed rats after spinal cord injury. The system mainly conducted image segmentation and coding of animal behavior through images taken using a high-speed camera. Parameters such as variation of joint angles, variances of gait cycle, stride length, and shift in center of gravity when walking, etc. were quantized through a pre-designed image analysis software program. Afterwards, the correlation between quantization parameters and traditional BBB scores was studied. This study also found that the variation of joint angle and the variance of the shift in center of gravity when walking have a good correlation with traditional assessment methods of BBB scores. Lastly, it is anticipated that the quantitative assessment system developed could provide an objective and reliable secondary assessment basis for future BBB score-related tests.
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