RESUMO
Tissue engineering leads to the development of biomaterial scaffolds where its biocompatibility and bioactivity are often improved after performing physical or chemical surface modification treatments. Micropatterning, soft lithography, and biofabrication are also approaches that provide a biomimetic microenvironment but have proven very costly and time consuming. In this concern, an appropriate substrate with suitable sites for cell attachment represents a major factor in cell behavior and biological functions. For this reason, our strategy was to fabricate a standard fibrous biomaterial with reproducible surface topography, incorporating microbeads and nanofeatures, and show the positive outcomes of the new substrate reflected on cell functions of bone cells. The electrospun polycaprolactone (PCL) beads-on-string membranes were obtained by adjusting the spinning solution at different concentrations until continuous beads were formed. Cell adhesion and proliferation, on the PCL scaffold, were analyzed the subsequent 2 days after initial culture. Complementary studies of cytoskeleton spreading and differentiation were analyzed after 7 and 14 days of the initial incubation. The scanning electron microscopy (SEM) images showed evidence of the formation of beads-on-string nanofibers and suggested that as-formed microstructures worked as attachment sites for osteoblasts. We investigated cell proliferation using anti-BrdU fluorescence assay, and results show a similar proliferation rate of cells cultured between PCL scaffolds and control. Finally, Phalloidin TRITC and antisialoprotein antibody were used to analyze cell spreading and differentiation after 7 and 14 days, respectively. This work shows a low-cost fabrication method to produce a biodegradable scaffold with micro/nanostructured characteristics that favor cell adhesion, proliferation, maturation, and subsequent differentiation of osteoblasts. According to the results, the biocompatibility of PCL beads-on-string could be comparable to other complex biomaterials, and we conclude that our scaffold is optimal for applications in bone tissue regeneration.
RESUMO
In this work, we explore the use of electrochemical methods (i.e., impedance) along with the arsenic-specific aptamer (ArsSApt) to fabricate and study the interfacial properties of an arsenic (As(III)) sensor. The ArsSApt layer was self-assembled on a gold substrate, and upon binding of As(III), a detectable change in the impedimetric signal was recorded because of conformational changes at the interfacial layer. These interfacial changes are linearly correlated with the concentration of arsenic present in the system. This target-induced signal was utilized for the selective detection of As(III) with a linear dynamic range of 0.05-10 ppm and minimum detectable concentrations of ca. 0.8 µM. The proposed system proved to be successful mainly because of the combination of a highly sensitive electrochemical platform and the recognized specificity of the ArsSApt toward its target molecule. Also, the interaction between the ArsSApt and the target molecule (i.e., arsenic) was explored in depth. The obtained results in this work are aimed at proving the development of a simple and environmentally benign sensor for the detection of As(III) as well as in elucidating the possible interactions between the ArsSApt and arsenic molecules.
RESUMO
Interfacial surface properties, both physical and chemical, are known to play a critical role in achieving long-term stability of cell-biomaterial interactions. Novel bone tissue engineering technologies, which provide a suitable interface between cells and biomaterials and mitigate aseptic osteolysis, are sought and can be developed via the incorporation of nanostructured materials. In this sense, engineered nanobased constructs provide an effective interface and suitable topography for direct interaction with cells, promoting faster osseointegration and anchoring. Therefore, herein we have investigated the surface functionalization, biocompatibility, and effect of cellulose-nanodiamond conjugates on osteoblast proliferation and differentiation. Cellulose nanocrystals (CNC) were aminated through a 3-aminopropyltriethyoxysilane (APTES) silylation, while nanodiamonds (ND) were treated with a strong acid oxidation reflux, as to produce carboxyl groups on the surface. Thereafter, the two products were covalently joined through an amide linkage, using a common bioconjugation reaction. Human fetal osteoblastic cells (hFOB) were seeded for 7 days to investigate the in vitro performance of the cellulose-nanodiamond conjugates. By employing immunocytochemistry, the bone matrix expression of osteocalcin (OC) and bone sialoprotein (BSP) was analyzed, demonstrating the viability and capacity of osteoblasts to proliferate and differentiate on the developed composite. These results suggest that cellulose-nanodiamond composites, which we call oxidized biocompatible interfacial nanocomposites (oBINC), have the potential to serve as a biointerface material for cell adhesion, proliferationand differentiation because of their osteoconductive properties and biocompatibility; furthermore, they show promising applications for bone tissue regeneration.
