Nanoscale implant anchorage aided by cement line deposition into titanium dioxide nanotubes.

The success of titanium dental implants relies on osseointegration, the load-bearing connection between bone tissue and the device that, in contact osteogenesis, comprises the deposition of bony cement line matrix onto the implant surface. Titanium dioxide nanotubes (NTs) are considered a promising surface for improved osseointegration, yet the mechanisms of cement line integration with such features remains elusive. Herein, we illustrate cement line deposition into NTs on the surface of titanium implants with two underlaying microstructures: a machined surface or a blasted/acid etched surface placed in the tibiae of Wistar rats. After retrieval, scanning electron microscopy of tissue reflected from the implant surface indicated minimal incursion of the cement line matrix into the NTs. To investigate this further, focused ion beam was utilized to prepare cross-sectional samples that could be characterized using scanning transmission electron microscopy. The cement line matrix covered NTs regardless of underlying microstructure, which was further confirmed by elemental analysis. In some instances, cement line infiltration into the NTs was noted, which reveals a mechanism of nanoscale anchorage. This study is the first to demonstrate cement line deposition into titanium NTs, suggesting nano-anchorage as a mechanism for the success of the NT modified surfaces in vivo.

Relative contributions of implant hydrophilicity and nanotopography to implant anchorage in bone at Early Time Points.

OBJECTIVE: To compare the contributions of implant hydrophilicity and nanotopography on anchorage in bone. The effect of elevated calcium surface chemistry on bone anchorage was also investigated. MATERIALS AND METHODS: A full factorial study design was implemented to evaluate the effects of ultraviolet (UV) light and/or sodium lactate (SL) and discrete crystalline deposition of nanocrystals (DCD) treatments on the osseointegration of dual acid-etched (AE) titanium alloy (Ti6Al4V) and grit blasted and AE (BAE) commercially pure titanium (CpTi) implants. Sodium hydroxide (NaOH)-treated CpTi implants were immersed in simulated body fluid (SBF) to increase calcium surface chemistry. Implants were placed in the femora of Wistar rats and tested using pull-out testing (BAE implants: 5, 9, 14 days) or tensile testing (AE implants: 9 days, NaOH implants: 28 days). RESULTS: Ti6Al4V-AE implants with DCD- and UV-treated surfaces significantly increased bone anchorage compared with untreated Ti6Al4V-AE alloy implants. Pull-out testing of BAE-CpTi implants with the DCD treatment showed increased disruption force values compared with surfaces without the DCD treatment at 5, 9 and 14 days by 4.1N, 13.9N and 15.5N, respectively, and UV-treated implants showed an increase at 14 days by 8.4N. No difference was found between NaOH + SBF and NaOH + H2 O groups. CONCLUSIONS: Bone anchorage of implants was found to be improved by UV-treating implants or nanotopographically complex surfaces. However, implant nanotopography was found to have a greater contribution to the overall bone anchorage and is more consistent compared with the time-dependent nature of the UV treatment.

Synergistic effect of polyelectrolyte multilayers and osteogenic growth medium on differentiation of human mesenchymal stem cells.

Layer-by-layer assembly of biogenic polyelectrolytes (PEL) was carried out on the surface of poly (L-lactide) to generate polyelectrolyte multilayers (PEM) that foster osteogenic differentiation of human mesenchymal stem cell (hMSC). Gelatin (GEL), hyaluronic acid (HA) and heparin (HEP) were chosen as polyanions, while chitosan (CHI) was employed as polycation. Multilayer formation was monitored by surface plasmon resonance and water contact angle measurements showing that layer formation process and surface wetting properties depended on the type of polyanions. While HEP as strong PEL led to thicker and more hydrophilic PEM, layer mass was lower for weak polyanions GEL and HA. Short-term adhesion studies with hMSC showed strong adhesion and spreading of cells on PEM composed of GEL/CHI and low spreading, motile phenotype and aggregation of hMSC on HEP/CHI or HA/CHI. Long term studies over three weeks were carried out to follow growth and differentiation of hMSC on the PEM. Weak osteogenic differentiation of hMSC was observed on GEL/CHI if cells were cultured in normal medium while no osteogenic phenotypes were observed on HEP/CHI or HA/CHI. When cells were cultured in osteogenic differentiation medium, however, PEM composed of HEP/CHI or HA/CHI promoted differentiation of hMSC towards osteoblasts, while PEM composed of GEL/CHI failed to do so. Overall, the composition of PEMs can be used as additional tool to control osteogenic differentiation of hMSC.

Biocompatibility of poly(L-lactide) films modified with poly(ethylene imine) and polyelectrolyte multilayers.

Poly(L-lactide) (PLLA) films were modified with poly(ethylene imine) (PEI) either by adsorption or covalent binding to prepare the material for immobilization of polyelectrolyte multilayers (PEM). Two different PEI, low- and high-molecular-weight (LMW or HMW, respectively) PEI, were used. The PEI modification efficiency was monitored via surface amino group density, water contact angle and X-ray photoelectron spectroscopy (XPS) measurements. Covalent binding of HMW PEI by a two-step-activation method produced the highest amino group density and the lowest water contact angle. On the other hand, the adsorption method resulted in moderate amounts of immobilized PEI on the surface. Subsequently sulphated hyaluronan and chitosan were used to form PEM on PLLA that was covalently modified with HMW PEI. Regular formation of PEM was achieved, which was demonstrated by change of water contact angles and mass increase measured with quartz crystal microbalance. An osteoblast-like cell line, MG 63, was used to test the effects of modifications on biocompatibility. Contrarily to earlier reports showing that particularly HMW PEI had certain cytotoxicity, it was found that all modifications including PEM resulted in a better biocompatibility than plain PLLA indicated by a more spread phenotype of cells, their increased growth and metabolic activity.

Immobilization of poly (ethylene imine) on poly (L-lactide) promotes MG63 cell proliferation and function.

Poly (ethylene imine) (PEI) is a polycation widely used for DNA transfection to cells but also applied as primary polycation for layer-by-layer (LBL) assembly of polyelectrolytes. The aim of the present study was to investigate the effect of modification with PEI on the biocompatibility of poly (L-lactide) (PLLA) films. PEI with different molecular weight was immobilized on PLLA by either adsorption or covalent binding. Cell morphologies, immuno-fluorescence staining, cell proliferation by lactate dehydrogenase assay and cell differentiation by alkaline phosphatase assay were utilized to assess the biocompatibility of the modified PLLA using osteoblast cell line MG63. Results revealed that PEI modification remarkably improved cell adhesion, viability, proliferation and function compared with plain PLLA. Hence, PEI-modified PLLA is acceptable as transfection vehicle for engineering of bone and other tissues, or as primary layer to allow LBL assembly to generate biomimetic surface coatings.

Recognition mechanism of theophylline-imprinted polymers: two-dimensional infrared analysis and density functional theory study.

Molecular imprinting polymers (MIPs) are synthetic materials having specific cavities tailored for a target molecule. Thoroughly understanding the molecular recognition mechanism is favorable for the rational design, preparation, and application of MIPs. In this work, theophylline (THO)-imprinted poly(acrylonitrile-co-acrylic acid) (PANCAA) films with acrylic acid (AA) as the functional monomer were fabricated and a set of concentration-dependent Fourier transform infrared (FT-IR) spectra were collected. Two-dimensional (2D) correlation analysis of the spectra and density functional theory (DFT) calculation were conducted to evaluate the molecular recognition mechanism. DFT at the B3LYP/6-31+G(d,p) level is efficacious to calculate the binding energies (DeltaE) and the theoretical vibration frequencies for the possible configurations of THO_AA complexes. An optimized cyclic hydrogen-bonded configuration (complex THO_AA1) has the highest binding energy (-16.63 kcal mol(-1)) that is more stable than others. In addition, the experimental vibrations of the carbonyl groups in the FT-IR spectra were assigned on the basis of the DFT results. Moreover, methylacrylic acid (MAA) and caffeine (CAF) as compared analogues were also investigated. The DFT-based theoretical predictions are coincident with the reported results.

Chitosan-modified poly(acrylonitrile-co-acrylic acid) nanofibrous membranes for the immobilization of concanavalin A.

Lectin affinity membranes have been receiving much attention for the separation and detection of various glycoconjugates. In this work, we present a simple and efficient method for the preparation of lectin affinity nanofibrous membranes. Chitosan-modified poly(acrylonitrile-co-acrylic acid) (PANCAA) nanofibrous membranes were first prepared by a coupling reaction between the primary amino groups of chitosan and the carboxyl groups of PANCAA electrospun membranes. Surface characterizations by attenuated total reflectance Fourier transform infrared spectroscopy (FT-IR/ATR), X-ray photoelectron spectroscopy (XPS) and field-emission scanning electron microscopy (FESEM) confirm the chemical and morphological changes of the studied nanofibrous membranes. Fluorescence-labeled concanavalin A (FL-Con A) was then immobilized on these membranes via noncovalent binding. Analyses by fluorescence spectrophotometer (FS) and confocal laser scanning microscopy (CLSM) reveal that the immobilization of Con A onto the modified nanofibrous membranes has been successfully achieved on the basis of the electrostatic interaction and the specific recognition between Con A and chitosan. The results show that the amount of adsorbed FL-Con A increases dramatically with the increasing coupling degree of chitosan (CDC) on the nanofibrous membrane. Moreover, Con A immobilized on the chitosan-modified nanofibrous membranes (CMNMs) can remain relatively stable at pH 5.3. Therefore, it is believed that this work may provide a new kind of material for affinity application.