In-situ polymerized polypyrrole nanoparticles immobilized poly(ε-caprolactone) electrospun conductive scaffolds for bone tissue engineering

Abstract

Despite intensive attempts to fabricate polypyrrole nanoparticles (PPy-NPs) incorporated nanofibrous scaffolds, a low-cost facile strategy is still demanded. Herein, we developed a novel strategy- in-situ polymerization of PPy-NPs and immobilized them into the PCL polymeric matrix in a single step. For the in-situ polymerization of PPy-NPs, ferric chloride hexahydrate (FeCl₃.6H₂O) was introduced as an oxidant into the blended solution of PCL and pyrrole monomers. Due to the chemical oxidative polymerization process, the clear solution changed into a black PCL/PPy solution. After electrospinning the solution, PCL/PPy composite nanofibers were fabricated. The immobilization of PPy-NPs into PCL matrix was clearly revealed by Bio-TEM images. The Field emission scanning electron microscopy (FESEM) results exhibited that the PCL/PPy scaffolds showed significantly decreased fiber diameter. The atomic force microscopy (AFM) study showed increased surface roughness in the PCL/PPy scaffolds. The mechanical strength test of PCL/PPy scaffolds showed improved Young's Modulus (YM = 2 to 4-folds) and tensile strength (TS = 3 to 4-folds). As well as the YM and TS were gradually increased with increased concentration of PPy-NPs in composite scaffolds. The conductivity measurement conducted on polymeric solution and electrospun scaffolds showed an increasing trend of conductive property in the PCL/PPy solution and scaffolds too. The surface wettability test exhibited decreased water contact angle measurement from 126° for pure PCL to 93° for the PCL/PPy-200 composite scaffold. The biomineralization test conducted by simulated body fluid (SBF) incubation showed enhanced calcium-phosphate crystal deposition on the PCL/PPy scaffolds. The CCK-8 assay and confocal laser scanning microscopic (CLSM) imaging conducted without and with electrical stimulation (ES) displayed enhanced cell adhesion, growth, and proliferation of MC3T3-E1 cells on the PCL/PPy conductive scaffolds. Furthermore, ALP and ARS staining assays showed significant enhancement of the calcium-phosphate deposition on the PCL/PPy scaffolds after ES treatment. Hence, the current study provides a novel strategy for the fabrication of PCL/PPy conductive scaffolds with enhanced bioactivity, biocompatibility, and osteogenic differentiation under electrical stimulation confirmed its promising application towards bone tissue engineering.

Keywords: Bone tissue engineering; Conductive scaffold; Electrical stimulation; Polycaprolactone; Polypyrrole.