J Periodontal Implant Sci.  2011 Apr;41(2):73-78.

Effect of seeding using an avidin-biotin binding system on the attachment of periodontal ligament fibroblasts to nanohydroxyapatite scaffolds: three-dimensional culture

• Jang YJ
• Jung IH
• Park JC
• Jung UW
• Kim CS
• Lee YK
• Kim CK
• Choi SH

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%. 1x10(5) 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.

Keyword

Cell adhesion; Fibroblasts; Periodontal ligament; Tissue engineering

MeSH Terms

Avidin
Biotin
Cell Adhesion
Dihydroergotamine
Fibroblasts
Humans
Microscopy, Electron, Scanning
Nanoparticles
Periodontal Ligament
Polymethyl Methacrylate
Porosity
Seeds
Tissue Engineering
Tooth
Avidin
Biotin
Dihydroergotamine
Polymethyl Methacrylate

Figure

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  Figure 1 WST-1 values. Each value reflects a ratio of attached living cells. a)Significant difference between the static and avidin biotin binding system (ABBS) groups, P<0.05. b)Significant difference between the agitating and ABBS groups, P<0.05.

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  Figure 2 Nanohydroxyapatite scaffold (×50).

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  Figure 3 Avidin-modified nanohydroxyapatite scaffold (×50).

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  Figure 4 Scanning electron micrographs of periodontal ligament fibroblasts attached to an nanohydroxyapatite scaffold. No meaningful difference was observed among the seeding methods: static cell seeding (A), agitating cell seeding (B), and avidin biotin binding system cell seeding (C, D) (×2,000).

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