| The stress urinary incontinence (SUI) is a disease that the urine flow outward without orders when the patients have a sudden pressure to their abdomen such as cough,laugh,cry,sneeze,raising heavy body. SUI mainly relate to the females in which the rate is about 15 percent to 60 percent, which especially give a lot of trouble to the old women. The SUI would be paid attention to with the ageing increasing on our society.The injection therapy has the virtues of safety, simple, convenience, less wound and easily practicing in out-patient service. The medicine or chemical reagents were injected into the mucosa and muscle around the internal os near the urinary bladder to lengthen and lessen the urethra. The resisting force of urinary can be increased and control the stress urinary incontinence successfully. At present the ideal material has not been discovered which have the virtues of better longer effect, lower cost and safety. The idea material should have some characteristics as following: no immunogenicity, no exclusion, lower anaphylaxis,nonvenomous,effctive,the same anatomical structure as the receptor organs, having less inflammation in the site of injecting and no shifting from the injecting site to other organs,easily practicing and reasonable cost. The mesenchymal stem cells (MSCs) not only have all above virtues but also are simple, cheaper, not moving away to other organs. The MSCs derived from the bone marrow can differentiated into osteoblasts, chondrocytes, lipocytes, myoblasts and neurocytes and so on. It is easy to isolate and culture MSCs in vitro. Because of lower immunogenicity to the receptors MSCs have be regarded as a better injection reagent.The main aim of this research is to observe the result of injecting mesenchymal stem cells or it's induced cells by 5-azacytidine into the the mucosa and muscle around the internal os near the urinary bladder of the SD rats which have the stress unitary incontinence and the effection on the SUI by the urine kinetics check and evoking sneeze experiment.The main experimental methods and results are as follows: Part 1: Study on isolating and culturing of the MSCs. (1) Selecting the concentration of FBS: analyzing the proliferation ability of MSCs with MTT method effected by the different concentrations of FBS including 0%, 5%, 10%, 15% and 20%, selecting 10% and 15% as the best experiment's result at last.(2) Selecting the density of the inoculating: analyzing and observing the proliferation state of MSCs derived from the bone marrow with MTT method effected by the different density of the inoculating including 2,4,6,8,10,12,14 and 16×105/ml, selecting 8 and 10×105/ml as the best density of the inoculating .(3) Observing the morphorgeny of MSCs: MSCs are well-distributed. Most of MSCs look like short poles or shuttles, meantime a little of them have anomalo-morphorgeny. There are growing like cloing. (4) Observing the transfer of culture MSCs: The transfer of MSCs pass the residence time, exponential growth time and plateau time.The first to the fouth generation of the MSCs proliferate quickly and their form is uniform which is more fit to be used to study on the induction and injection therapy.Part 2: Study on inducing MSCs to differentiate into myoblasts cells by 5-Aza-CR. (1) Selecting the concentration of 5-Aza-CR: analyzing the proliferation ability of MSCs with MTT method ,induced by the different concentrations of 5-Aza-CR including 0,1,3,5,10 and 20umol/L, selecting 5 and 10μmol/L as the best concentration of 5-Aza-CR.(2) Observing the morphorgeny changes of induced MSCs: MSCs enlarge and look like fat-haulm at the 10th day after induced by 10umol/L 5-Aza-CR.There are cells like myotube and fusing phenomena between two to three cells at the 13th day and myotube structure observed at the 15-17th day.(3) Investigating protein expression of myoblasts: some MSCs begin to express desmin on the 8th day after induced by 10μmol/L 5-Aza-CR; The expression degree enhances on the 15th day.Some MSCs begin to express myosin on the...
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