塗佈細胞外間質蛋白質在乙烯-乙烯醇共聚物上對於誘導 真皮乳頭微組織形成的影響

Abstract

Biomaterials are a very important field in tissue engineering. It has been shown that biomaterials can affect the morphology of cells. For certain cells, specific cellular function is determined by its intercellular organization. To be specific, the hair follicle (HF) induction activity of dermal papilla (DP) cells can only be preserved when they are kept in an aggregated state or microtissues. We have demonstrated that DP cells can spontaneously grow into spheroidal microtissues on poly(ethylene-co-vinyl-alcohol) (EVAL) and these microtissues are able to induce HF neogenesis. However, DP cells have a compromised attachment on EVAL, and this may lead to cell loss during culture. In this work, we try to improve the cell attachment on EVAL by coating various extracellular matrix proteins to enhance cell attachment with the aim at more efficient production of DP microtissues on EVAL. The purpose of this research is to investigate the attachment, migration and aggregation of DP cells into microtissue on EVAL which is coated with various extracellular matrix proteins including fibronectin, collagen I, collagen IV and laminin. We found that cell attachment is enhanced by the coating of all the proteins tested. However, DP microtissue formation is only observed on the fibronectin-coated surface. We compare the cell mobility on the extracellualr matrix protein-coated surface and found that cell migration rate is reduced by the coating of collagen I, collagen IV and laminin while cells are maintained in a highly motile state on fibronectin-coated surface. In the case of the fibronectin-coated surface, there is a 66% increase of DP microtissue formation compared with that of non-coated EVAL. The increase in cell attachment and DP microtissue formation on the fibronectin-coated surface can be abolished by the treatment of fibronectin antibody. The result indicates that, in addition to cell-substrate adhesivity, cell migration is also a major determinant of microtissue formation. Each extracellualr matrix protein has different effects on DP cells attachment and migration on biomaterials. Cells can only grow into DP microtissues when the extracellualr matrix proteins can keep them in a more motile state. Overall, fibronectin-coating can enhance DP microtissue formation on EVAL by increasing cell-substrate adhesivity without compromising cell motility. Fibronectin-coating can improve the efficiency of DP microtissue production and can be applied to tissue engineering for HF regeneration.