Surface modification of titanium thin film with chitosan via electrostatic self-assembly technique and its influence on osteoblast growth behavior

Chitosan (Chi) and poly (styrene sulfonate) (PSS) were employed to surface modify titanium thin film via electrostatic self-assembly (ESA) technique in order to improve its biocompatibility. The surface chemistry, wettability and surface topography of the coated films with different number of deposited layers were investigated by using X-ray photoelectron spectroscopy (XPS), water contact angle measurement and atomic force microscopy (AFM), respectively. The results indicated that a full surface coverage for the outmost layer was achieved at least after deposition of five layers, i.e., PEI/(PSS/Chi)₂ on the titanium films. The formed multi-layered structure of PEI(PSS/Chi) _x (x ≥ 2) on the titanium film was stable in air at room temperature and in phosphate buffered solution (PBS) for at least 3 weeks. Cell proliferation, cell viability, DNA synthesis as well as differentiation function (alkaline phosphatase) of osteoblasts on chitosan-modified titanium film (PEI/(PSS/Chi)₆) and control sample were investigated, respectively. Osteoblasts cultured on chitosan-modified titanium film displayed a higher proliferation tendency than that of control (p < 0.01). Cell viability, alkaline phosphatase as well as DNA synthesis measurements indicated that osteoblasts on chitosan-modified titanium films were greater (p < 0.01) than those for the control, respectively. These results suggest that surface modification of titanium film was successfully achieved via deposition of PEI/(PSS/Chi) _x layers, which is useful to enhance the biocompatibility of the titanium film.     

Effects of baicalin-modified poly(D,L-lactic acid) surface on the behavior of osteoblasts

In the present study, the functions of rat calvaria osteoblasts on baicalin-modified poly(D,L-lactic acid) (PDLLA) films were investigated in vitro. The surface characteristics of surfaces (both modified and control) were investigated by water contact angle measurement and electron spectroscopy for chemical analysis (ESCA). Cell morphologies on these surfaces were examined by scanning electron microscopy (SEM). Cell adhesion and proliferation were used to assess cell growth on the modified and control surfaces. The MTT assay was used to determine cell viability and alkaline phosphatase (ALP) activity was performed to evaluate differentiated cell function. Compared to control films, cell attachment of osteoblasts on baicalin-modified PDLLA film was significantly higher (P<0.05 and P<0.01) after 6 h and 8 h culture, and cell proliferation was also significantly greater (P<0.05 and P<0.01) at the end of 4th and 7th day, respectively. The MTT assay suggested that the cell viability of osteoblasts cultured on baicalin-modified PDLLA film was significantly higher (P<0.05) than that seeded on the control. Meanwhile, the ALP activity of osteoblasts cultured on modified films was also considerably enhanced (P<0.01) compared to that found on control. These results revealed that the biocompatibility PDLLA could be improved by surface modification with baicalin.     

Modulation of poly (d,l-lactic acid) with amniotic fluid for improvement of the cell affinity

The objective of this study was to investigate the effects of naturally occurring amniotic fluid modified poly(d,l-lactic acid) (PDLLA) film on the culture of rat calvaria osteoblast. The characteristics of surfaces (both modified and control) were examined by contact angle measurement and electron spectroscopy for chemical analysis (XPS). Cell adhesion and proliferation were used to assess the cell behavior on modified films and control one. MTT assay was used to determine cell viability and alkaline phosphatase (ALP) activity was taken to evaluate differentiated cell function. Compared with the untreated films, cell adhesion of osteoblast was significantly higher (P < 0.05) than that found on control, and osteoblast proliferation was also greater than control one (P < 0.01) at the time interval of 4 and 7 days. Moreover, the alkaline phosphatase (ALP) activity exhibited statistic difference (P < 0.05) and cell viability demonstrated significant difference (P < 0.01) between amniotic fluid modified PDLLA films and control one. These results suggested that amniotic fluid was a suitable material when used to modify PDLLA in order to improve its biocompatibility.     

Surface modification of poly-L-lactic acid films by electrostatic self-assembly to promote vascular smooth muscle cells growth

The intention of this study was to surface modify the poly-L-lactic acid (PLLA) film and evaluate the effects of the surfaces on the growth of vascular smooth muscle cells (VSMCs) in vitro. Collagen and hyaluronic acid (HA) were utilized as polycation and polyanion in this study. Layer-by-layer (LBL) self-assembly technique was used to lead to the formation of multilayer moleculer on the poly-L-lactic acid (PLLA) film surfaces. Collagen/HA layers was overcasted coating on the PLLA surface after the activation layers by poly-(ethyleneimine) (PEI). The structure and morphology of the multilayer molecular were examined by attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectrophotometer and atomic force microscope (AFM), respectively. The ATR-FTIR analysis illuminated the presence of collagen on the PLLA surface. The AFM results showed the multilayer appeared on PLLA surface. The VSMCs were adopted to evaluate the cyto-compatibility of the modified PLLA films. It was found that the viability of VSMCs on the modified PLLA films were greater than that on original PLLA films and tissue culture plastic after ten days culture (p < 0.05). Scanning electron microscopy (SEM) and laser scanning confocal microscopy (LSCM) data also confirmed the homogeneous results. These data suggest that collagen/HA coat can be successfully adopted in the surface modification of PLLA film through LBL technique, and also can enhance its cell compatibility.     

Histological study of surface modified three dimensional poly (<Emphasis Type="SmallCaps">d</Emphasis>,<Emphasis Type="SmallCaps"> l</Emphasis>-lactic acid) scaffolds with chitosan in vivo

Biocompatibility and tissue regenerating capacity are essential for biomaterials that used in tissue engineering. The aim of this study was to histologically assess the tissue reactions and bone conductivities of surface modified three dimensional (3-D) poly (d, l-lactic acid) (PDLLA) scaffolds, which were coated with chitosan via a physical entrapment method. The native PDLLA scaffold was prepared via thermally induced phrase separation technique and was characterized by scanning electron microscopy (SEM) and differential scanning calorimetry (DSC). Osteocalcin assay, a method to evaluate the bone formation potential, has shown that the osteocalcin production in chitosan-modified 3-D PDLLA scaffold group was significantly higher (p < 0.05) than that of in control. The tissue reactions and bone conductivities between surface modified PDLLA and native PDLLA scaffolds were evaluated using a rabbit radialis defect model in vivo and compared at different implantation intervals (2, 4, 8 and 12 weeks). The histological results have shown a higher bone formation potential and better biocompatibility of chitosan-modified 3-D PDLLA scaffolds as compared with the control group scaffolds.     

Formation and surface modification of self-assembled films with acrylated hyperbranched poly(ester-amine) as the outmost layer

An acrylated hyperbranched poly(ester-amine) (HPEA) synthesized from piperazine and trimethylolpropanetriacrylate at a molar ratio of 1:1.42 was used as the polycation to form self-assembled films by layer-by-layer dipping with poly(sodium-p-styrenesulfonate) as the polyanion. The surface morphology and hydrophilicity of the films with HPEA as the outmost layer were controlled by adjusting the solution pH of HPEA. Due to the existence of many acrylate groups, the films with HPEA as the outmost layer were further reacted with a series of reagents, including piperazine, piperidine, laurylamine and p-phenylenediamine. The surface reactions of the films depended on both the nature of the reagents and the morphology of the initial surface. In the presence of the strong basic amines, piperazine and piperidine, the films dissolved extensively into the solution. In the case of laurylamine, a modified film with an opaque appearance was obtained due to the crystallization of long alkyl chains. With the weak basic amine p-phenylenediamine as the reactant, the film was effectively modified without any change in the surface morphology.     

Biocompatibility and bioactivity of PDLLA/TiO2 and PDLLA/TiO2/Bioglass® nanocomposites

Biocompatibility and bioactivity of polymer matrix composites containing titanium dioxide (TiO₂) nanoparticles were investigated. The solvent casting method was used to prepare poly (d,l-lactic acid) (PDLLA) films with 0 and 20 wt.% TiO₂ nanoparticles and with 20 wt.% TiO₂ mixed with 5 wt.% micrometre-sized (< 5 μm) Bioglass® particles. The samples were characterized using scanning electron microscopy (SEM), X-ray diffraction (XRD) and energy Dispersive X-ray (EDX) analyses. A Zygo® light interferometer was used to examine the surface roughness of the samples. The bioactivity and the surface reactivity of the materials were determined by investigating the formation of hydroxyapatite (HA) on the surface of samples upon immersion in simulated body fluid (SBF) for up to 28 days. Heterogeneous distributed HA crystals were found on composite films containing TiO₂ after 21 days exposure to SBF. Cell cytotoxicity and viability were determined by using live/dead and MTS assay on osteoblast-like MG-63 cells. The PDLLA films containing different concentrations of TiO₂ and Bioglass® particulate inclusions showed no effect on cell viability in live/dead assay after incubation period of 7 days. All three groups of samples demonstrated significant increase in relative metabolic activity in MTS assay after 7 days incubation (while a slower proliferation rate was obtained for cells on the PDLLA film containing both TiO₂ and Bioglass® compared to the Thermanox® control). The bioactive behaviour of the nanocomposites may make them attractive materials for fabrication of tissue engineering scaffolds.     

In vitro behavior of layer-by-layer deposited molecular oligoelectrolyte films on Ti–6Al–4V surfaces

Layer-by-layer self-assembled films of molecular oligoelectrolytes were used to modify Ti–6Al–4V surfaces in order to test their ability as potential drug delivery system. With regard to medical application the in vitro behavior of the modified material was investigated. The Ti–6Al–4V (6% aluminium, 4% vanadium) material was treated in a layer-by-layer (LbL) process with 2, 4, 6 and 8 layers of molecular oligoelectrolytes 1 and 2 and thereby doped with a fluorescent reporter molecule 2. Human osteoblasts were cultured for a period up to 5 days on the modified material. Ti–6Al–4V surfaces without modification were used as control. In order to investigate the in vitro behavior of the coating as well as the influence of components of the coating on osteoblastic cells, respectively, cell proliferation, differentiation and attachment of hFOB cells were observed by means of cell number, osteoblastic gene expression and fluorescence microscopy. Degradation behavior of the OEM (oligoelectrolyte multilayer film) was examined using optical spectroscopy. Measurement data imply that the layer-by-layer coating was successfully assembled on the Ti surface and endures steam sterilization. The fluorescence signal in cell culture medium increased strictly linear with increasing pre-assembled number of layers on the surface. Proliferation rates of the cells in experimental groups did not differ significantly from each other (P ≥ 0.783). Differentiation pattern was not significantly changed by the coating. The fluorescent reporter component of the film was absorbed by osteoblastic cells and was detected by fluorescence microscopy.     

Titanium dioxide (TiO<Subscript>2</Subscript>) nanoparticles filled poly(<Emphasis Type="SmallCaps">d</Emphasis>,<Emphasis Type="SmallCaps">l</Emphasis> lactid acid) (PDLLA) matrix composites for bone tissue engineering

Titanium dioxide (TiO₂) nanoparticles were investigated for bone tissue engineering applications with regard to bioactivity and particle cytotoxicity. Composite films on the basis of poly(d,l lactid acid) (PDLLA) filled with 0, 5 and 30 wt% TiO₂ nanoparticles were processed by solvent casting. Bioactivity, characterised by formation of hydroxyapatite (HA) on the materials surface, was investigated for both the free TiO₂ nanoparticles and PDLLA/TiO₂ composite films upon immersion in supersaturated simulated body fluid (1.5 SBF) for up to 3 weeks. Non-stoichiometric HA nanocrystals (ns-HA) with an average diameter of 40 nm were formed on the high content (30 wt% TiO₂) composite films after 2 weeks of immersion in 1.5 SBF. For the pure PDLLA film and the low content composite films (5 wt% TiO₂) trace amounts of ns-HA nanocrystals were apparent after 3 weeks. The TiO₂ nanopowder alone showed no bioactivity. The effect of TiO₂ nanoparticles (0.5–10,000 μg/mL) on MG-63 osteoblast-like cell metabolic activity was assessed by the MTT assay. TiO₂ particle concentrations of up to 100 μg/mL had no significant effect on MG-63 cell viability.     

Multilayer films consisted of azulene-based dye molecules and polyelectrolyte: Preparation, characterization and photoluminescent property

Ultrathin multilayer films of two azulene-based (Az-based) dye molecules (Az), 3-methylazulene-1-carboxylic acid hydrazide (Az-1) and 5-(4-phenylamine)-2-(3-methylazulene-1-yl)-1,3,4-oxadiazoles (Az-2), and poly(sodium 4-styrenesulfonate) (PSS) have been prepared by layer-by-layer self-assembly and characterized by UV-vis spectroscopy, fluorescence spectroscopy, small-angle X-ray reflectivity measurements and atomic force microscopy (AFM) imaging. UV-vis spectra show that the characteristic absorbance values of the multilayer films increase almost linearly with the number of PSS/Az bilayers, suggesting that the deposition process is regular and highly reproducible from layer to layer. Average thicknesses for the PSS/Az-1 and PSS/Az-2 bilayers of the multilayer films are ca. 0.9 and 1.4 nm, respectively. AFM images provide the surface morphology of the PSS/Az films, indicating that the film surface is relatively uniform and smooth. The occurrence of photoluminescent activity conforms the potential for creating luminescent multilayer films with Az-based dye molecules.     

Colorimetric response of fluorescein-modified multilayer thin films induced by electrolysis of water

The surface of an indium–tin oxide (ITO) electrode was coated with layer-by-layer (LbL) thin films composed of fluorescein-modified poly(allylamine) (F-PAH) and poly(styrenesulfonic acid) (PSS) and their UV–visible absorption spectra were recorded under the influence of electrode potential. The LbL films were prepared by an alternate deposition of F-PAH and PSS on the surface of ITO electrode through an electrostatic force of attraction. The intensity of the absorption band around 500 nm originating from fluorescein residues in the LbL film depended on the pH of the solution in which the LbL film is immersed. The intensity of the absorption band decreased when the electrode potential higher than + 1.2 V was applied, while virtually no response was observed at lower electrode potential. The spectral change was suppressed in solutions with higher buffer capacity. The results were discussed in terms of the changes in local pH in the vicinity of the electrode surface, which in turn was induced by electrolysis of H₂O on the electrode surface.     

Effects of hydrolysis on dodecyl alcohol-modified bioactive glasses and PDLLA/modified bioactive glass composite films

In this article, mesoporous 58S and 58S bioactive glasses (BGs) were surface modified by dodecyl alcohol through esterification reaction and PDLLA/modified BGs composite films were prepared. The purpose of this study was to investigate the properties of the modified BGs particles and the PDLLA/modified BGs composite films before and after hydrolytic treatment. The modified BGs powders and composite films were treated in boiling water for 20 min to remove the dodecyl chains. After hydrolytic treatment, the modified BGs powders showed increased hydrophilicity and the FTIR analysis revealed that most dodecyl chains were removed. Furthermore, the hydrophilicity of the PDLLA/modified BGs composite films was also greatly improved. The tensile strength of the composite films after hydrolysis decreased slightly, but was still much higher than that of pure PDLLA film. In addition, bone marrow mesenchymal stem cells from dogs on the composite films after hydrolytic treatment showed the highest proliferation rate. The results suggest that hydrolytic treatment is an effective and practicable method to remove alcohol chains from surface-modified BGs and polymers/modified BG composites, which may be used for preparation of bioactive scaffolds for tissue engineering applications.     

Green isolation and physical modification of pineapple stem waste starch as pharmaceutical excipient

The waste of inedible parts of pineapple, particularly in tropical countries, contributes to environmental burden. This study aimed to utilize pineapple stem waste as a source of starch-based pharmaceutical excipient. The starch was isolated from pineapple stem waste using a simple process without applying harsh chemicals. The isolated starch (PSS) was then physically modified through gelatinization and spray drying to improve its physical properties. Starch characteristics were identified by FTIR, TGA, and XRD analysis. The SEM imaging showed morphological change with reduced surface roughness due to physical modification of the starch. Decreased crystallinity of modified starch (MPS) was confirmed by our XRD results: the peaks of A-type crystalline at 2θ of 13°, 15°, 18°, and 23° were present in PSS, yet mostly absent in MPS. Thermogravimetric analysis showed that MPS behaved differently from PSS and the degradation events occurred at lower temperature. When the starch was spray-dried without prior gelatinization process, the physicochemical characteristics of spray-dried starch resembled untreated starch. Moisture content in PSS (10.66%) decreased after gelatinization to 7.3%. Potential use of MPS was demonstrated by its powder flowability (Student’s t test, p?<?0.05), swelling capacity (Student’s t test, p?<?0.05), and compaction profile. In summary, our findings demonstrated that modified pineapple starch showed better physical characteristics and quite promising as a tablet binder and disintegrant.     

Adhesion and growth of vascular smooth muscle cells in cultures on bioactive RGD peptide-carrying polylactides

The surface of poly(l-lactide) (PLLA) films deposited on glass coverslips was modified with poly(dl-lactide) (PDLLA), or 1:4 mixtures of PDLLA and PDLLA-b-PEO block copolymers, in which either none, 5% or 20% of the copolymer molecules carried a synthetic extracellular matrix-derived ligand for integrin adhesion receptors, the GRGDSG oligopeptide, attached to the end of the PEO chain. The materials, perspective for vascular tissue engineering, were seeded with rat aortic smooth muscle cells (11,000 cells/cm²) and the adhesion, spreading, DNA synthesis and proliferation of these cells was followed on inert and bioactive surfaces. In 24-h-old cultures in serum-supplemented media, the number of cells adhering to the PDLLA-b-PEO copolymer was almost eight times lower than that on the control PDLLA surface. On the surfaces containing 5% and 20% GRGDSG-PEO-b-PDLLA copolymer, the number of cells increased 6- and 3-fold respectively, compared to the PDLLA-b-PEO copolymer alone. On PDLLA-b-PEO copolymer alone, the cells were typically round and non-spread, whereas on GRGDSG-modified surfaces the cell spreading areas approached those found on PDLLA, reaching values of 991 μm² and 611 μm² for 5% and 20% GRGDSG respectively, compared to 958 μm² for PDLLA. The cells on GRGDSG-grafted copolymers were able to form vinculin-containing focal adhesion plaques, to synthesize DNA and even proliferate in a serum-free medium, which indicates specific binding to the GRGDSG sequences through their adhesion receptors.     

Surface modification of titanium substrates with silver nanoparticles embedded sulfhydrylated chitosan/gelatin polyelectrolyte multilayer films for antibacterial application

To develop Ti implants with potent antibacterial activity, a novel “sandwich-type” structure of sulfhydrylated chitosan (Chi-SH)/gelatin (Gel) polyelectrolyte multilayer films embedding silver (Ag) nanoparticles was coated onto titanium substrate using a spin-assisted layer-by-layer assembly technique. Ag ions would be enriched in the polyelectrolyte multilayer films via the specific interactions between Ag ions and –HS groups in Chi-HS, thus leading to the formation of Ag nanoparticles in situ by photo-catalytic reaction (ultraviolet irradiation). Contact angle measurement and field emission scanning electron microscopy equipped with energy dispersive X-ray spectroscopy were employed to monitor the construction of Ag-containing multilayer on titanium surface, respectively. The functional multilayered films on titanium substrate [Ti/PEI/(Gel/Chi-SH/Ag) _n /Gel] could efficiently inhibit the growth and activity of Bacillus subtitles and Escherichia coli onto titanium surface. Moreover, studies in vitro confirmed that Ti substrates coating with functional multilayer films remained the biological functions of osteoblasts, which was reflected by cell morphology, cell viability and ALP activity measurements. This study provides a simple, versatile and generalized methodology to design functional titanium implants with good cyto-compatibility and antibacterial activity for potential clinical applications.     

Fabrication and characterization of nanostructured conducting polymer films containing magnetic nanoparticles

In this study, the layer-by-layer technique is used to deposit nanostructured films exhibiting electrical conductivity and magnetic behavior, from poly(o-ethoxyaniline) (POEA), sulfonated polystyrene (PSS) and positively-charged maghemite nanoparticles. In order to incorporate the nanoparticles into the films, maghemite nanoparticles, in the form of magnetic fluid, were added to POEA solutions, and the resulting suspensions were used for film deposition. UV-Vis spectroscopy and atomic force microscopy images reveal that POEA remains doped in the films, even in the presence of the maghemite nanoparticles, and its typical globular morphology is also present. Electrical measurements show that a POEA/PSS film prepared from POEA solution containing 800 µL of the magnetic fluid exhibits a similar conductivity to that of the control film and, additionally, magnetic measurements indicated that nanosized maghemite phase was incorporated within the polymeric film.     

等离子体处理偶联胶原提高聚乳酸材料的细胞相容性

本研究通过结合氨气等离子体处理以及胶原改性的方法来改善聚乳酸材料的细胞相容性。水接触角分析表明氨气等离子体处理可以改善聚乳酸材料的亲水性,促进细胞的粘附。光学显微镜以及扫描电镜的分析结果表明,氨气等离子体处理后通过胶原改性可以进一步促进细胞在聚乳酸材料表面的生长。并且,与涂覆胶原相比,偶联胶原可以更好地使胶原蛋白固定在聚乳酸材料表面,从而表现出更好的细胞相容性。     

Effects of hydrolysis on dodecyl alcohol modified β-CaSiO3 particles and PDLLA/modified β-CaSiO3 composite films

In this research, β-CaSiO₃ particles were surface modified with dodecyl alcohol, and Poly-(DL-lactic acid) (PDLLA)/modified β-CaSiO₃ composite films were fabricated with a homogenous dispersion of β-CaSiO₃ particles in the PDLLA matrix. The aim of the study was to investigate the properties of the composite films before and after hydrolytic treatment. SEM images showed retained homogenous dispersion of β-CaSiO₃ particles after hydrolysis and tensile test also showed maintained mechanical property. Simulated body fluid (SBF) incubation experiment suggested that hydrolytic treatment did not affect the formation of hydroxyapatite on the surface of the composite films. The hydrophilicity of the composites was greatly recovered (from 69.82° to 50.28°) after hydrolysis. In addition, cells cultured on composite films after hydrolysis presented the highest cell proliferation rate and differentiation level. All of these results suggested that the surface modification of silicate particles with dodecyl alcohol along with reversible hydrolytic treatment was an effective and feasible approach to fabricate polymer/silicate composite materials with improved properties.     

Electrical properties of thin PbTe films on Si substrates

An attempt was made to reduce the carrier concentration in thin PbTe films on Si substrates by optimizing deposition conditions. A modified hot-wall method was used for reproducible growth ofp-type films with 5 × 10¹⁵ < p(77 K) < 5 × 10¹⁷ cm^-3 andn-type films with 3 × 10¹⁵ < n(77 K) < 5 × 10¹⁶ cm^-3. The IR irradiation was found to have a significant effect on the temperature variation of film resistance. The activation energy of the IR-sensitivity centers was determined to be 0.11 ± 0.005 eV at room temperature and 0.18 ± 0.005 eV between 150 and 180 K.     

Nonaqueous preparation of nanocomposite films containing both polyoxometalate and phthalocyanine with bifunctional electrocatalytic properties

New bifunctional nanocomposite films containing water-insolvable cobalt tetraaminophthalocyanine (CoTAPc) and 2:18 tungstophosphate anions (P₂W₁₈) were successfully fabricated using the electrostatic layer-by-layer (LbL) assembled method in nonaqueous (DMF) media. UV–vis spectroscopy, IR spectroscopy, X-ray photoelectron spectroscopy, atomic force microscopy (AFM) and electrochemical method were used to characterize the assembled multilayer films. UV–vis measurements revealed regular film growth with each four-layer {P₂W₁₈/CoTAPc/PSS/PAH} adsorption. AFM analysis provides the morphology of the multilayer films. The cyclic voltammetric curves indicate that the film shows electrocatalytic activity for chlorate anion.