A new nanocomposite scaffold based on polyurethane and clay nanoplates for osteogenic differentiation of human mesenchymal stem cells in vitro

Abstract

Bone tissue engineering as an alternative strategy, provides a great opportunity for regeneration of large bone tissue lesions. The use of biodegradable porous scaffolds along with stem cells, cytokines and growth factors improves cell survival, adhesion, proliferation and differentiation. In the present study, clay nanoplates (CNPs) were surface-modified (MCNPs) using phosphoric acid and calcium hydroxide, then porous polyurethane (PU) scaffolds and PU-MCNPs nanocomposite scaffolds were synthesized using solvent evaporation-dissolution technique. Physicochemical and morphological properties of scaffolds and MCNPs were studied by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and scanning electron microscopy (SEM). Moreover, thermal behavior of scaffolds was assessed by differential scanning calorimetry (DSC). Degradability, water uptake and mechanical behaviors of scaffolds were evaluated and hydrophilicity properties of them were obtained by contact angle technique. MTT assay and Acridine Orange/Ethidium Bromide (AO/EB) staining were used to assess the biocompatibility of MCNPs and PU scaffolds regarding cell attachment and proliferation support. Osteogenic differentiation of cultured human adipose derived mesenchymal stem cells (hADSCs) on MCNPs, PU and PU-MCNPs scaffolds was evaluated using common osteogenic markers such as alkaline phosphatase (ALP) activity, calcium content assay, Alizarin Red staining, immunocytochemical analysis (ICC) and quantitative real-time PCR (qPCR). According to the results, the surface modification of CNPs and their presence into the PU scaffolds significantly enhanced proliferation and osteogenic differentiation of hADSCs. These results were obtained by higher ALP enzyme activity, biomineralization and expression of osteogenic related proteins and genes in differentiated hADSCs on PU-MCNPs scaffolds. In conclusion, our results revealed that these biocompatible nanocomposites porous scaffolds with proper cell adhesion and proliferation as well as effective osteogenic differentiation and which are able to provide a new and useful matrix for bone tissue engineering purposes.