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Nowadays, human dental pulp stem cells (hDPSCs) became more attractive for therapeutic purposes because of their high proliferation and differentiation potential. Thus, coupling the desired cellular characteristics of hDPSCs with good biomaterial properties of the chitosan scaffolds provide an interesting approach for tissue engineering applications. On the other hand, scaffold surface modification is also needed to promote stem cell adhesion since chitosan lacks adhesion motifs to support direct cell anchorage. In this study, hDPSCs were isolated from third molars of healthy female individuals (aged 16-25) with enzymatic digestion. For cell culture studies, the chitosan scaffolds which have approximately 9 mm diameter and 2 mm thickness with interconnected structure were prepared by freeze-drying. To support cellular attachment the scaffolds were covalently immobilized with either RGD (arginine-glycine-aspartic acid) or fibronectin (Fn) molecules. Cells were seeded on chitosan scaffolds with or without immobilized RGD and fibronectin. Cell attachment, spreading, adhesion behaviors and proliferation capacity were examined by scanning electron microscopy, immunofluorescence staining and PrestoBlue assays, respectively. In addition, differentiation potential of hDPSCs on Fn immobilized chitosan scaffolds was determined with real time reverse transcriptase polymerase chain reaction analysis. The results showed that chitosan scaffolds were not able to support stem cell attachment. hDPSCs on chitosan scaffolds formed spheroids more quickly and the size of spheroids were smaller than on chitosan-RGD while Fn-immobilized chitosan scaffolds strongly supported cellular attachment but not odontogenic differentiation. The results suggest that the Fn-immobilized chitosan scaffolds may serve as good three-dimensional substrates for dental pulp stem cell attachment and proliferation. In the case of dental regeneration, they must be supported by appropriate biosignals to induce odontogenic differentiation.

The aim of the present study is to eliminate the bioinertness of polyetheretherketone (PEEK) material and to increase its osteogenic activity by applying a number of surface modifications in order to discover the most effective method. First, the surface of the bare PEEK (B-PEEK) was mechanically modified by sandblasting (S-PEEK). As a second method, physical modification was provided by etching of B-PEEK in 10 M sodium hydroxide (NaOH) solution at 60 °C for 48 h (N-PEEK). Following the sandblasting, S-PEEK samples were also etched with NaOH to obtain SN-PEEK samples. In order to increase osteogenic activity of the mechanically and/or physically modified PEEK samples, they were coated with boron-doped nanohydroxyapatite (B-nHAp) in the presence of microwave energy. Thus, B-PEEK/B-nHAp, S-PEEK/B-nHAp, N-PEEK/B-nHAp and SN-PEEK/B-nHAp samples were obtained. While water contact angles and surface roughness of the B-PEEK samples increased after modification, hydrophilicity increased with B-nHAp coating. Cell culture results demonstrated that high proliferation and differentiation capacities of MC3T3-E1 cells were obtained on the B-nHAP-coated surfaces compared to uncoated specimens. However, channeled texture of both the N-PEEK and N-PEEK/B-nHAp samples enhanced Col1a1 mRNA expression and collagen secretion in addition to increased alkaline phosphatase (ALP) activity when compared to other groups. As a result, it was determined that the bioactivity increased on the modified PEEK surfaces, but the most effective osteogenic activity was provided with B-nHAp coatings. The synergetic effect of NaOH etching and B-nHAp coatings might be used as a promising surface modification method to use PEEK material in orthopedic and dental applications with increased osteogenic activity.

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