In Vitro Study Of PLGA Nano-fiber Scaffold With Sustained Release Of Plasmid BFGF And BMP-2 In Promoting Periodontal Ligament Cells' Proliferation And Differentiation

Objective: Poly(lactic-co-glycolic acid)(PLGA)nanofiber scaffolds with the encapsulation of plasmid bFGF(basic fibroblast growth factor,bFGF)and BMP-2(bone morphogenetic protein,BMP-2)was prepared by tissue engineering technology,when transfected into hPDLCs these two factors were released slowly.The purpose of this study was to observe the effects of PLGA on the proliferation and differentiation of hPDLCs,and to provide an experimental basis for periodontal ligament regeneration after the replantation of an avulsed tooth.Materials and Methods: The PLGA nanofiber scaffolds and pbFGF/pBMP-2-PLGA composite scaffolds were prepared by electrospinning.The surface morphology of scaffolds was characterized by SEM and TEM.The third passage of periodontal ligament cells was seeded on the surface of the scaffold material.The confocal fluorescence microscopy was used to detect the cell adhesion.MTT were used to detect the cell proliferation.And qRT-PCR were used to detect the osteogenic differentiation related genes(Runx-2,ALP)and fibroblast differentiation related genes(SCX,Col-I).The results were analyzed by SPSS 17.0 software,data of two sample means were analyzed using the Independent-Samples T Test,differences were statistically significant when P < 0.05.Results: Scanning electron microscopy showed that both of PLGA fibrous scaffolds and pbFGF/pBMP-2-PLGA composite nanofiber scaffolds are smooth and uniform,non-bead and spindle structure.The average fiber diameter of the PLGA nanofiber was 417.21 ±70.68 nm,and the pbFGF/pBMP2-PLGA composite nanofiber was 263.96 ± 41.44 nm.The results of gene release showing that: The total release of pbFGF and pBMP-2 at 21 days was 128 ng.Confocal fluorescence microscopy showed that both of two scaffolds favored cell adhesion and growth.MTT results showed that the composite scaffold significantly promoted cell proliferation on day 7(P < 0.05).At the same time,qRT-PCR showed that SCX,Col-I,Runx-2 and ALP gene expression in the composite scaffold group was higher than of PLGA nano-fiber scaffold group(P < 0.05).Conclusions: The pbFGF/pBMP-2-PLGA composite nanofiber scaffolds can promote the proliferation of periodontal ligament cells,and osteogenic/fibroblast differentiation ability is better than PLGA nanofiber scaffold,which has the potential to promoteperiodontal tissue regeneration.