BMP-2 immobilized PCL/P3ANA nanofibers for bone tissue engineering

Abstract

Poly (e-caprolactone)/poly (m-anthranilic acid) (PCL/P3ANA) nanofibers were fabricated by electrospinning. Nanofibers were biofuctionalized with Bone morphogenic protein-2 (BMP-2) with 1-ethyl-3-(dimethyl-aminopropyl) carbodiimide hydrochloride (EDC)/N-hydroxyl succinimide (NHS) surface activation of carboxyl groups (-COOH) on PCL/P3ANA scaffold. The structure of nanofibers was investigated by FTIR-ATR spectroscopy. The amount of bound protein was determined by BCA protein assay. In vitro cell culture studies were conducted by using human osteocarsinoma cell line (SAOS-2) and protein immobilized PCL/P3ANA nanofibers as tissue engineering scaffolds. Cell viability and proliferation assays were performed in order to understand the cell behaviour against PCL/P3ANA nanofibers. Fluorescence staining of F-actin was performed to observe the cell morphology. Alkaline phoshotase (ALP) activity of SAOS-2 cells were investigated. The results confirmed the successful immobilization of BMP-2 on the PCL/P3ANA nanofibers which can be used in bone tissue engineering.