Corneal Stromal Tissue Engineering Using Keratocyte Spheroids Fabricated on High Molecular Weight Hyaluronic Acid Coating

• Main Authors: I Hao Tu, 涂益豪
• Other Authors: J. Y. Lai
• Format: Others
• Published: 2011
• Online Access: http://ndltd.ncl.edu.tw/handle/24180121198404268757

碩士 === 長庚大學 === 生化與生醫工程研究所 === 99 === Hyaluronic acid for the extracellular matrix (extracellular matrix, ECM) of the important biological components and high biocompatibility. Therefore, the biomedical material has been widely used in ophthalmology. Purpose of this study is to explore the corneal stromal cells cultured in different molecular weight hyaluronic acid coating of the behavior. The results showed that the molecular weight of hyaluronic acid with a more negative charge is increased, high surface roughness and the more hydrophilic properties. Light microscopic observations showed that the keratocyte adhesion decreased with increasing surface roughness of HA-modified substrates. The weak cell-matrix interaction and strong cell-cell interaction also contributed to the formation of multicellular spheroids on these nanostructured surfaces. Light microscopic observations showed that the keratocyte adhesion decreased with increasing surface roughness of HA-modified substrates. The weak cell-matrix interaction and strong cell-cell interaction also contributed to the formation of multicellular spheroids on these nanostructured surfaces. Cell proliferation and ECM production studies indicated that the keratocytes with a spheroid configuration were mitotically quiescent and exhibited a better biosynthetic capacity. Irrespective of surface nanotopographies of HA coating, the cells have good viability, suggesting that the keratocyte spheroids with a suitable size did not develop hypoxia-mediated cytotoxicity. Quantitative real-time reverse transcription polymerase chain reaction assays demonstrated that the keratocytes within spheroids showed enhanced phenotypic expression. Experimental evaluation of cell suspensions and cell spherical applied to corneal tissue engineering. Corneal thickness and intraocular pressure analysis showed that the cell spheroids to 14 days after treatment returned to normal cornea. As the cell spheroids secrete large amounts of extracellular matrix and then fill the damage of the stroma. Therefore, slit lamp showed that the cell spheroids mode of treatment can repair the scar tissue and it can be sustained in the continuous treatment of corneal stroma by Analysis of tissue sections stained. On the other hand, Analysis of the corneal light transmittance, the results revealed that cell spheroids can effectively treat corneal damage to the cornea returned to normal light penetration after 14 days. Our findings suggest that the corneal keratocytes cultured on HA coatings can sense nanotopographic ECM cues. Bioengineering of multicellular spheroid may provide a new strategy for potential corneal stromal regenerative medicine.