碩士 / 國立臺灣科技大學 / 化學工程系 / 102 / The studies of treatments for articular cartilage defects have received lots of attentions due to the limitations of self-repair abilities of articular cartilage.
Cyanophycin, a non-ribosomal synthesis protein, has good biocompatibility and biodegradability, and its structure is positively charged which is helpful for cell adhesion. Dextran, a kind of natural polysaccharides, also has good biocompatibility and biodegradability in addition to good water solubility and good capability for water retention. However, it has no charges for cells to adhere onto. In the previous studies regarding tissue regeneration, dextran has been used as a substitute for glycosaminoglycan, and combined with other cell-adhesion polymers to mimic the structure of extracellular matrix for cell growth. This study is based on “biomimetic”, to combine the above two biomaterials, and focused on assessing the growth of porcine chondrocytes on the films of cyanophycin-dextran conjugate.
The grafting ratios of cyanophycin-dextran conjugate were determined by colorimetry, gel permeation chromatography (GPC) and elemental analysis (EA), and Fourier transform infrared spectroscopy (FT-IR) was used to analyze the change in the functional groups of the conjugates. The films of cyanophycin-dextran conjugate were cross-linked with glutaraldehyde, and the cross-linking results were observed under a fluorescence microscopy after fluorescence staining. The swelling ratios were tested at different time intervals. CHO cells were grown on the films as a primary test for cell viability, and then two different passages of porcine chondrocytes were further grown on the films. The cell viability was monitored by MTT assay, Bradford protein assay and DNA quantitation. The glycosaminoglycan contents were examined by Blyscan assay, and the specific mRNA expressions were measured by qRT-PCR.
The results showed that the grafting of dextran enhanced the water solubility of cyanophycin, and the swelling ratios of the films increased with dextran grafting. On the other hand, the grafting of cyanophycin improved cell-adhesion on the polysaccharides, one of the major limitations of cartilage self-repair. Meanwhile, moderate contents of cyanophycin could maintain the advantages of dextran which provided a growth condition for chondrocytes similar to an in vivo one. Moreover, the films could be helpful for aging chondrocytes.
In conclusion, the films of cyanophycin-dextran conjugate could maintain cell viability and delay chondrocyte dedifferentiation, and could help aging chondrocytes regenerate.
| Identifer | oai:union.ndltd.org:TW/102NTUS5342087 |
| Date | January 2014 |
| Creators | Sheng-Yang, Chen, 陳陞陽 |
| Contributors | Wen-Chi Tseng, 曾文祺 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 214 |
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