Effect of seeding using an avidin-biotin binding system on the attachment of periodontal ligament fibroblasts to nanohydroxyapatite scaffolds: three-dimensional culture
장용주, 정임희, 박정철, 최성호, 정의원, 김창성, 이용근, 김종관,
소속 상세정보
장용주 ( Jang Yong-Ju ) - Yonsei University College of Dentistry Department of Periodontology
정임희 ( Jung Im-Hee ) - Yonsei University College of Dentistry Department of Periodontology
박정철 ( Park Jung-Chul ) - Yonsei University College of Dentistry Department of Periodontology
최성호 ( Choi Seong-Ho ) - Yonsei University College of Dentistry Department of Periodontology
정의원 ( Jung Ui-Won ) - Yonsei University College of Dentistry Department of Periodontology
김창성 ( Kim Chang-Sung ) - Yonsei University College of Dentistry Department of Periodontology
이용근 ( Lee Yong-Keun ) - Yonsei University College of Dentistry Department of Dental Biomaterials and Bioengineering
김종관 ( Kim Chong-Kwan ) - Yonsei University College of Dentistry Department of Periodontology
KMID : 0363020110410020073
Abstract
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.
키워드
Cell adhesion; Fibroblasts; Periodontal ligament; Tissue engineering
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