Objectives: The purpose of the present study was to examine the growth and organizational behavior of a potential allergenic graft cell on chitosan membrane that were with or without extracellular proteins (fibronectin or collagen I coating). We view this as an important initial step in developing of salivary gland tissue engineering. Methods: We fabricate chitosan polymer membrane by the method of dry processing. To investigate the biocompatibility of membranes, we used cell culture method and MTT assays were performed for measurement of cell viability. To evaluate the situation of cell attachment, the cells on chitosan membranes were investigated by scanning electron microscopy (SEM). Utilizing indirect immunofluorescence analysis, cells were shown to express cytokeratin, rat parotid acinar cell specific secretory protein. Immunocytochemical staining of the acinar cells involved the exposure to specific antibodies for the cytokeratins. Results: Acinar cells have been difficult to maintain in primary or secondary cultures over extended periods of time. We have developed a long-term culture method for acinar cells growth. The rat parotid acinar cell adhesion was found to increase with collagen I coated chitosan membranes. We also found sialic acid is important for promoting the activity of parotid acinar cells. Conclusion: Salivary gland loss due to trauma, post irradiation damage due to oral cancer or a variety of genetic disorders such as autoimmune exocrinopathy Sjogren's syndrome continues to affect most adults adversely in their lives. We have initiated a program to develop an artificial salivary gland for patients suffered from salivary gland disorder. .A long term culture method for rat parotid acinar cells was developed and cell growth on the chitosan membrane with collagen coating was better than that without coating.

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