Attachment ability of periodontal ligament fibroblast on avidin-biotin binding system : 3-dimensional culture

Abstract

[한글]

[영문]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 function. Limited contact frequency between cells and scaffolds and inadequate time for effective cell adhesion have been observed using conventional 2-dimensional cell seeding onto 3-dimensionally reconstructed scaffolds. Some methods have been reported of seeding cells to 3-dimensional scaffold and the authors have asserted its usefulness and effectiveness. One of the examples is avidin and biotin molecular pair. Avidin-biotin binding system (ABBS) is distinguished for its extraordinary high affinity and has been extensively applied to biotechnology. The aim of this study is to explore the cell attachment of periodontal ligament (PDL) fibroblasts on nano hydroxyapatite (n-HA) scaffold using ABBS.Human periodontal ligament fibrobasts 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 3mm in height disk with high (81.55%) porosity. 1 x 105 PDL fbroblasts were applied to each scaffold. The cells were seeded into scaffolds by static, agitating and ABBS seeding method. The number of PLD fibroblasts attached was greater for ABBS seeding method than for static or agtiting method (p<0.05). Typical examples of SEM images show the morphology of PDL fibroblasts adhered to the scaffolds. No meaningful difference has been observed among seeding methods. However, increased strength of cell attachment of ABBS could be deduced from the high affinity between avidin and biotin (Kd = 10-15 M). In conclusion, we demonstrated that avidin modified n-HA scaffold could enhance the attachment of biotinylated PDL fibroblasts.