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KMID : 0363020110410020073
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Journal of Korean Academy of Periodontology
2011 Volume.41 No. 2 p.73 ~ p.78
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Effect of seeding using an avidin-biotin binding system on the attachment of periodontal ligament fibroblasts to nanohydroxyapatite scaffolds: three-dimensional culture
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Jang Yong-Ju
Jung Im-Hee
Park Jung-Chul
Choi Seong-Ho
Jung Ui-Won
Kim Chang-Sung
Lee Yong-Keun
Kim Chong-Kwan
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Abstract
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Purpose: For periodontal tissue engineering, it is a primary requisite and a challenge to select the optimum types of cells, properties of scaffold, and growth factor combination to reconstruct a specific tissue in its natural form and with the appropriate function. Owing to fundamental disadvantages associated with using a two-dimensional substrate, several methods of seeding cells into three-dimensional scaffolds have been reported and the authors have asserted its usefulness and effectiveness. In this study, we explore the cell attachment of periodontal ligament fibroblasts on nanohydroxyapatite (n-HA) scaffold using avidin biotin binding system (ABBS).
Methods: Human periodontal ligament fibroblasts were isolated from the health tooth extracted for the purpose of orthodontic procedure. HA nanoparticles were prepared and Ca(NO3)2-4H2O and (OC2H5)3P were selected as precursors of HA sol. The final scaffold was 8 mm in diameter and 3 mm in height disk with porosity value of 81.55%. 1×105 periodontal ligament fibroblasts were applied to each scaffold. The cells were seeded into scaffolds by static, agitating and ABBS seeding method.
Results: The number of periodontal ligament fibroblasts attached was greater for ABBS seeding method than for static or agitating method (P<0.05). No meaningful difference has been observed among seeding methods with scanning electron microscopy images. However, increased strength of cell attachment of ABBS could be deduced from the high affinity between avidin and biotin (Kd=10-15 M).
Conclusions: The high-affinity ABBS enhances the ability of periodontal ligament fibroblasts to attach to three-dimensionally constructed n-HA scaffolds.
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KEYWORD
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Cell adhesion, Fibroblasts, Periodontal ligament, Tissue engineering
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