Analysis of Electrospun Nanofibrous Scaffolds as Matrices for Gingival Reconstruction

Objective: Sub- epithelial connective tissue grafting remains the gold standard for treatment of gingival recession, but it involves harvesting of tissue from a second surgical site and the amount of graft tissue is limited. In recent years, attention has turned to the field of regenerative medicine for alternate strategies for generation of gingival grafts. We outline here our initial in vitro experiments in which we assessed human gingival fibroblasts (HGFs) growth and differentiation on polycaprolactone (PCL) electrospun fibres loaded with basic fibroblast growth factor (bFGF) microspheres.

Method: The scaffold was fabricated by simultaneously electrospinning PCL fibers and electrospraying bFGF-containing microspheres. HGFs were seeded onto scaffolds and grown for 24 h, 1 and 2 weeks. Cell attachment and spreading was assessed using scanning electron microscopy. Proliferation was assessed using CyQuant assays. Cell differentiation was quantified using Taqman Realtime PCR with primers specific to collagen type 1 and a-smooth muscle actin (a-SMA).

Result: HGFs attached and spread on all scaffolds with 24 hrs and by 7 days had formed multilayered cell sheets on the scaffolds. HGFs could be seen to penetrate the outlayer of the scaffold, with significant ingrowth to space between the fibers evident. HGFs had significantly higher mRNA levels of both collagen type I and a-SMA compared with control scaffolds. 

Conclusion: PCL electrospun fibrous scaffolds containing bFGF-containing microspheres enhance cell attachment, proliferation and differentiation of HGFs. We conclude that these scaffolds could serve as a matrix to enhance gingival regeneration.