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Introduction: The main aim of tissue engineering is to design biological substitutes in order to enhance and restore damaged tissues or organs, and biomaterials for fabricating the scaffolds play a crucial role. Collagen (COL), the main part of the ECM and a group of structural proteins which contributes to the extracellular scaffolding, was considered for scaffolds preparation. Studies have shown that incorporation of functionalized multi-walled carbon nanotubes (MWCNTs) into various polymeric scaffolds, including COL, can enhance their mechanical, physicochemical, and biological properties.. Several pre-clinical, and clinical studies have shown that curcumin (CUR) has antioxidant, anti-inflammatory, and anti-tumor effects. However, poor bioavailability of CUR owing to decreased serum content, reduced tissue absorption, elevated metabolic rate, and fast removal from the body hampered its practical use. Therefore, incorporation of CUR into scaffolds can enhance its bioavailability at the implantation site. In this study, a novel biomimetic tri-dimensional (3D) scaffolds containing COL (extracted from rat tail), MWCNTs, and CUR were prepared by freeze-drying technique. It was aimed to increase the mechanical properties of the scaffolds by MWCNTs and to promote the anti-inflammatory, bioactivity, and biocompatibility properties of those scaffolds by incorporation of CUR.
Materials and Methods: 0.5 to 1.5% MWCNTs and 5 to 15% CUR were added to the pure COL solutions and freeze-dried. Mechanical characterization of the scaffolds was evaluated by a universal testing machine. Physical and chemical characterization of the scaffolds were also assessed by scanning electron microscopy (SEM) and Fourier-transform infrared spectroscopy (FTIR). Biological characterization of the scaffolds was also evaluated by in-vitro bioactivity for four weeks, in-vitro biodegradability and the rate of CUR release for 8 weeks, in-vitro biocompatibility using rat synovial-derived mesenchymal stem cells (SMSCs) for seven days, and in-vivo biocompatibility for six weeks.
Results: SEM revealed highly interconnected porosity of the obtained 3D structures with porosity from 75% to 85%.. The addition of up to 1% MWCNTs and 10% CUR has enhanced the mechanical properties of the scaffolds. The presence of MWCNTs and CUR in the COL scaffolds was confirmed by FTIR. The developed COL-MWCNTs composite scaffolds containing 10% CUR revealed excellent in-vitro cytotoxicity and cell attachment using SMSCs. After 8 weeks of examination, the rate of CUR release into the PBS buffer was 20%. Importantly, scaffolds modified with CUR showed reduced inflammatory reactions in the rat model after six weeks of implantation.
Conclusion: CUR incorporated into COL-MWCNTs scaffolds has let to higher mechanical properties, has enhanced the formation of HA crystals on the surface of the scaffold, and has promoted in vitro and in vivo biocompatibility of the scaffolds by making better interaction between the scaffolds and SMCSs and also by reducing the inflammatory reaction. Overall, the newly developed COL-MWCNTs-CUR 10% freeze-dried scaffolds have demonstrated their high potential for tissue engineering applications.
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