Mesenchymal stem cells (MSCs) and osteoblasts respond to the surface electrical charge and topography of biomaterials. This work focuses on the connection between the roughness of calcium phosphate (CP) surfaces and their electrical potential (EP) at the micro- and nanoscales and the possible role of these parameters in jointly affecting human MSC osteogenic differentiation and maturation in vitro. A microarc CP coating was deposited on titanium substrates and characterized at the micro- and nanoscale. Human adult adipose-derived MSCs (hAMSCs) or prenatal stromal cells from the human lung (HLPSCs) were cultured on the CP surface to estimate MSC behavior. The roughness, nonuniform charge polarity, and EP of CP microarc coatings on a titanium substrate were shown to affect the osteogenic differentiation and maturation of hAMSCs and HLPSCs in vitro. The surface EP induced by the negative charge increased with increasing surface roughness at the microscale. The surface relief at the nanoscale had an impact on the sign of the EP. Negative electrical charges were mainly located within the micro- and nanosockets of the coating surface, whereas positive charges were detected predominantly at the nanorelief peaks. HLPSCs located in the sockets of the CP surface expressed the osteoblastic markers osteocalcin and alkaline phosphatase. The CP multilevel topography induced charge polarity and an EP and overall promoted the osteoblast phenotype of HLPSCs. The negative sign of the EP and its magnitude at the micro- and nanosockets might be sensitive factors that can trigger osteoblastic differentiation and maturation of human stromal cells.
Fourier transform infrared microscopy was used to investigate human cortical bone samples before and after treatment with increasing doses of X-ray radiation. Especially the spectral region of the v1 and v3 phosphate vibrations of hydroxyapatite, the main mineral component of bone, and the region of the amide I and amide II vibrational bands due to the collagen extracellular matrix were examined. Major spectral changes in the phosphate region between 1250–1000 cm–1 occur after irradiation doses between 1 and 4 Gray. These findings are explained by a decrease in size of mineral crystallites and by variances of the stoichiometric/non-stoichiometric apatite composition. The Ca2+/PO4 3-/HPO4 2- composition in the biological apatite is altered near the bone surface. The secondary structure of the collagen matrix is not affected by cumulative irradiation up to doses of 15 Gray as indicated by the unchanged frequency maximum and contour shape of the amide I band between 1600–1700 cm–1. However, side chain carboxylate groups of the collagen matrix that are involved in coordination with apatite bound calcium ions are partially removed by decarboxylation upon irradiation. Concomitantly, a loss of acidic phosphate groups due to a formation of phosphate groups with bound calcium is observed. These changes on a molecular level can be correlated with alterations in the mechanical properties of the bone samples, e.g. with an increased embrittlement as deduced from experiments with a scanning acoustic microscope (1).
This article is focusing on electrical functionalization of biomaterial’s surface to enhance its biocompatibility. It is an overview of previously unpublished results from a series of experiments concerning the effects surface electrical functionalization can have on biological systems. Saccharomyces cerevisiae cells were used for biological experiments. The hydroxyapatite (HAp) specimens were used to investigate influence of structural point defects on the surface electrical charge. Threshold photoelectron emission spectroscopy was used to measure the electron work function of HAp and biologic samples. The density functional theory and its different approximations were used for the calculation of HAp structures with defects. It was shown that the electrical charge deposition on the semiconductor or dielectric substrate can be delivered because of production of the point defects in HAp structure. The spatial arrangements of various atoms of the HAp lattice, i.e., PO4 and OH groups, oxygen vacancies, interstitial H atoms, etc., give the instruments to deposit the electrical charge on the substrate. Immobilization of the microorganisms can be achieved on the even surface of the substrate, characterized with a couple of nanometer roughness. This cells attachment can be controlled because of the surface electrical functionalization (deposition of the electrical charge). A protein layer as a shield for the accumulated surface charge was considered, and it was shown that the protein layer having a thickness below 1 µm is not crucial to shield the electrical charge deposited on the substrate surface. Moreover, the influence of surface charge on the attachment of microorganisms, when the surface roughness is excluded, and the influence of controlled surface roughness on the attachment of microorganisms, when surface charge is constant, were also considered.
The electronic structure and self-polarization of P(VDF-TrFE) Langmuir-Blodgett nanofilms were analyzed under temperature-driven phase transitions, according to their thickness, composition, and structural conformation. Both thermo-stimulated exoelectron emission (TSEE) spectroscopy and computational simulation, including quantum-chemical calculations from first principles, were carried out. PVDF and composite P(VDF-TrFE) (70:30) molecular chains as Trans and Gauche conformers, as well as crystal cells, were modeled for these TSEE analyses. The quantum-chemical calculations and the computational simulation were based on the density functional theory (DFT) as well as semi-empirical (PM3) methods. It was demonstrated that the energy of electron states, as well as the total energies of the studied P(VDF-TrFE) molecular clusters during phase transformation, is influenced by electron work function and electron affinity. Analysis was performed by combining TSEE experimental data with the computational data of the molecular models, demonstrating the effectiveness of this joint approach. For the first time, TSEE was used for contactless measurements of nanofilm polarization, and characterization of the phase transition. The proposed new method can be widely applied in nanobiomedicine, particularly in development of new bone bio-implants, including built-in sensors (new smart nanotechnology).
Advances in nanotechnology and nanomaterials have facilitated the development of silicon dioxide, or Silica, particles as a promising immunological adjuvant for the generation of novel prophylactic and therapeutic vaccines. In the present study, we have compared the adjuvanting potential of commercially available Silica nanoparticles (initial particles size of 10–20 nm) with that of aluminium hydroxide, or Alum, as well as that of complete and incomplete Freund's adjuvants for the immunisation of BALB/c mice with virus-like particles (VLPs) formed by recombinant full-length Hepatitis B virus core (HBc) protein. The induction of B-cell and T-cell responses was studied after immunisation. Silica nanoparticles were able to adsorb maximally 40% of the added HBc, whereas the adsorption capacity of Alum exceeded 90% at the same VLPs/adjuvant ratio. Both Silica and Alum formed large complexes with HBc VLPs that sedimented rapidly after formulation, as detected by dynamic light scattering, spectrophotometry, and electron microscopy. Both Silica and Alum augmented the humoral response against HBc VLPs to the high anti-HBc level in the case of intraperitoneal immunisation, whereas in subcutaneous immunisation, the Silica-adjuvanted anti-HBc level even exceeded the level adjuvanted by Alum. The adjuvanting of HBc VLPs by Silica resulted in the same typical IgG2a/IgG1 ratios as in the case of the adjuvanting by Alum. The combination of Silica with monophosphoryl lipid A (MPL) led to the same enhancement of the HBc-specific T-cell induction as in the case of the Alum and MPL combination. These findings demonstrate that Silica is not a weaker putative adjuvant than Alum for induction of B-cell and T-cell responses against recombinant HBc VLPs. This finding may have an essential impact on the development of the set of Silica-adjuvanted vaccines based on a long list of HBc-derived virus-like particles as the biological component.
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