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Cell Journal، جلد ۱۵، شماره Suppl ۱، صفحات ۳۴-۳۴
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عنوان انگلیسی Ps-32: Survey Specifics of Nucleus Pulposus Cells of Human Intervertebral Disc in Chitosan-Gelatin and Alginate Scaffolds
چکیده انگلیسی مقاله Objective: Low back pain is a major social problem nowadays. Intervertebral disc herniation and central degeneration of disc are two major reasons of low back pain that occur because of structural impairment of discs. Intervertebral disc includes the annulus fibrosus, transitional region, and nucleus pulposus (NP). NP forms the central nucleus of the disc. Reduction of cell count and extracellular matrix, especially in NP, causes disc degeneration.have been used for tissue repairing and regeneration of intervertebral disc in tissue engineering. Most scaffolds have biodegradable and biocompatible characteristics and also prepare a fine condition for proliferation and migration of cells. Beacause degeneration is result of decrease ECM and aggrecan is important proteoglycan in NP tissue of IVD thus goal of this study was survey and comparison rate of secreted aggrecan,morphology of NP cells and proliferation and viability of NP cells in alginate and chitosan-gelatin scaffolds. Materials and Methods: NP cells were derived by enzymatic hydrolysis of collagenase from NP tissue of patients undergoing open surgery for discectomy in Alzahra Hospital (Isfahan, Iran). Chitosan was blended with gelatin and glutaraldehyde was used for cross linking of the two polymers. Then, alginate scaffold was prepared. After approving the NP cells by flow cytometry of cytokeratin 18 marker, a cellular suspension with 4 × 105 cells was transferred to each scaffold and cultured for 21 days. Cell viability and proliferation were investigated by trypan blue and methyl thiazolyl tetrazolium (MTT) assay. A scanning electron microscope (SEM) was used to assert the porosity and to survey the structures of the scaffolds. ElISA assay used for determination of aggrecan secretion by NP cells seeded in chitosan-gelatin and alginate scaffolds. Results: We can use flow cytometry of cytokeratin 18 markers for recognition of NP cells. 80% of NP cells express CK18. MTT assay demonstrated that cell viability on the third day had significant difference with the first day in both scaffolds. There was also a significant reduction in cellular viability from day 3 to day 21. ELISA assay showed had significant difference with the first day in both scaffolds. There was also a significant increase in produced of aggrecan from day 3 to day 21 (p< 0.001). Alginate scaffold had higher secretion of aggrecan from day 3 ot day21. Results of cell count showed that mean difference between cell counts in alginate scaffold was significantly more than chitosan-gelatin scaffold (p< 0.001). Conclusion: Flowcytometry of cytokeratin 18 can be used as a method for recognition of NP cells. Compared to chitosan-gelatin scaffold, alginate scaffold prepared a better condition for proliferation of NP cells, maintain of morphology of cells and produce of aggrecan. The results of this study suggested that alginate scaffold could be useful in in-vivo studies and treatment. Different scaffolds (natural and synthetic)
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نشانی اینترنتی http://celljournal.org/journal/article/abstract/837
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