Nowadays, despite significant advances in the field of biomaterials for tissue engineering applications, novel bone substituents still need refinement so they can be successfully implemented into the medical treatment of bone fractures. Generally, a scaffold made of synthetic polymer blended with nanofillers was proven to be a very promising biomaterial for tissue engineering, however the choice of components for the said scaffold remains questionable. The objects of the presented study were novel composites consisting of poly(ε-caprolactone) (PCL) and two types of graphene materials: graphene oxide (GO) and partially reduced graphene oxide (rGO). The technique of choice, that was used to characterize the obtained composites, was Raman micro-spectroscopy. It revealed that the composite PCL/GO differs substantially from the PCL/rGO composite. The incorporation of the GO particles into the polymer influenced the structure organisation of the polymeric matrix more significantly than rGO. The crystallinity parameters confirmed that the level of crystallinity is generally higher in the PCL/GO membrane in comparison to PCL/rGO (and even in raw PCL) that leads to the conclusion that the GO acts as a nucleation agent enhancing the crystallization of PCL. Interestingly, the characteristics of the studied nanofillers, for example: the level of the organisation (D/G ratio) and the in-plane size of the nano-crystallites (La) almost do not differ. However, they have an ability to influence polymeric matrix differently.
Modern medical treatment can be improved by nanotechnology methods for preparing nanocomposites with novel physical, chemical and biological properties. The materials studied and analysed as membranes were produced from poly(ε-caprolactone) (PCL), which contained identical amounts of nano-additives, either montmorillonite (MMT) or functionalized multi-walled carbon nanotubes (MWCNT-f), while the reference membranes were obtained from unmodified PCL. In addition to the conventional methods used in the study of materials for medical purposes such as DSC, contact angle measurements, surface topography, Raman spectroscopy was also applied. Raman microspectroscopy can decode the phenomenon that occurs in the polymer in contact with the nanoparticles. Besides identifying the vibrations of certain functional groups, the calculation of crystallinity parameters is also possible, by which the most intense interactions within the nanocomposites can be analysed. The Raman studies indicate that each of the nano-additives reacts differently with the polymer matrix, which results in material properties that influence its biological properties. MWCNT-f interacts preferentially with the oxygen-containing groups, and particularly with the backbone regions in the vicinity of the single CO bond. The human osteoblast-like MG-63 cells, cultured on the PCL/MWCNT-f membrane for three days, show almost 100% viability.
Poly(ε-caprolactone) (PCL) is a biocompatible resorbable material, but its use is limited due to the fact that it is characterized by the lack of cell adhesion to its surface. Various chemical and physical methods are described in the literature, as well as modifications with various nanoparticles aimed at giving it such surface properties that would positively affect cell adhesion. Nanomaterials, in the form of membranes, were obtained by the introduction of multi-walled carbon nanotubes (MWCNTs and functionalized nanotubes, MWCNTs-f) as well as electro-spun carbon nanofibers (ESCNFs, and functionalized nanofibers, ESCNFs-f) into a PCL matrix. Their properties were compared with that of reference, unmodified PCL membrane. Human osteoblast-like cell line, U-2 OS (expressing green fluorescent protein, GFP) was seeded on the evaluated nanomaterial membranes at relatively low confluency and cultured in the standard cell culture conditions. The attachment and the growth of the cell populations on the polymer and nanocomposite samples were monitored throughout the first week of culture with fluorescence microscopy. Simultaneously, Raman microspectroscopy was also used to track the dependence of U-2 OS cell development on the type of nanomaterial, and it has proven to be the best method for the early detection of nanomaterial/cell interactions. The differentiation of interactions depending on the type of nanoadditive is indicated by the ν(COC) vibration range, which indicates the interaction with PCL membranes with carbon nanotubes, while it is irrelevant for PCL with carbon nanofibers, for which no changes are observed. The vibration range ω(CH2) indicates the interaction for PCL with carbon nanofibers with seeded cells. The crystallinity of the area ν(C=O) increases for PCL/MWCNTs and for PCL/MWCNTs-f, while it decreases for PCL/ESCNFs and for PCL/ESCNFs-f with seeded cells. The crystallinity of the membranes, which is determined by Raman microspectroscopy, allows for the assessment of polymer structure changes and their degradability caused by the secretion of cell products into the ECM and the differentiation of interactions depending on the carbon nanostructure. The obtained nanocomposite membranes are promising bioactive materials.
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