碩士 / 國立臺北科技大學 / 化學工程與生物科技系化學工程碩士班 / 107 / At present, most new pathogens have anti-drug factors, which cause antibiotics to fail to achieve the expected antibacterial effect, so phage therapy is gradually gaining attention. In recent years, many studies have covered wounds with wound dressing, temporarily replacing the function of epidermal or endothelial tissue protection, and promoting wound healing and recovery of damaged tissues. In order to protect the affected part from bacterial infection, the phage was grafted onto the sodium alginate film with calcium ions to prepare a lyophilized film carrying phage as a bacteriostatic dressing, which is expected to have a bacteriostatic effect on the wound dressing.
In this experiment, a phage capable of lysing Escherichia coli (DH5α) was isolated from seawater, a sodium alginate film was prepared by freeze-drying method, and an aqueous calcium chloride solution and a phage were mixed as a crosslinking agent to graft the phage onto the sodium alginate film. To investigate the activity and bacteriostatic ability of phage on the membrane. The experimental part was used to quantify and bacteriostatic test on the phage on the film, and the distribution and appearance of E. coli on the surface of the film were taken by scanning electron microscope. In addition, fibroblasts (L929) were inoculated on the surface of the phage and phage-free membranes. The DNA content and the appearance of the cell surface on the surface of the film were taken by scanning electron microscopy.
The results showed that each membrane was grafted (9.07±1.22)×104 PFU phage. The antibacterial test showed that the number of E. coli bacteria on the phage-containing samples was significantly less than that of the phage-free samples (p<0.05), and was observed under a scanning electron microscope. Escherichia coli on the phage membrane was ruptured, and the number of bacteria was less than that of the phage-free membrane. In terms of cytotoxicity, the DNA content of the cells in the phage-containing group was significantly higher than that in the phage-free group (p<0.01). The tensile test results showed that there was no significant difference between the phage-containing group and the non-phage group (p>0.05). The degradation test results also showed that the phage did not affect the film degradation rate (p>0.05).
In this experiment, the phage was successfully grafted onto the sodium alginate film by calcium ions, and its activity and antibacterial ability were maintained. The grafted phage did not change the mechanical strength and degradation rate of the film, and did not cause toxicity to the cells. Taken together, the above results show that the phage can be combined with the substrate as a bacteriostatic dressing.
| Identifer | oai:union.ndltd.org:TW/107TIT0006A086 |
| Date | January 2019 |
| Creators | HONG, JIA-SHUN, 洪嘉勛 |
| Contributors | LIN, HSIN-YI, 林忻怡 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 86 |
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