Surface modification of zirconia ceramics through cold plasma treatment and the graft polymerization of biomolecules.

Background/purpose: Although zirconia ceramics were highly versatile as dental implants, their long-term presence in the human body may slow down healing and impede cell growth in the past. To enhance the cytocompatibility of zirconia ceramics, surface activation modification was used to immobilize biopolymers such that a biomimetic environment was created. Materials and methods: Hexamethyldisilazane thin films were deposited onto the surface of inorganic zirconia through cold plasma treatment under various power and deposition time settings to form an organosilane interface layer. Next, oxygen plasma treatment was performed to activate the free radicals on the surface. Subsequently, ultraviolet light was employed to graft and polymerize acrylic acid for generating carboxyl groups on the surface. This was followed by a condensation reaction with biopolymers (chitosan, chitosan/poly-γ-glutamic acid, and gelatin). Results: Under a 20-min deposition time at 40 W and 150 mTorr, the thin films had a maximum graft density of 2.1 mg/cm2. MG-63 cells (human osteosarcoma cells) were employed to evaluate cell compatibility. Chitosan and chitosan/poly-γ-glutamic acid promoted the compatibility of MG-63 cells (a human osteosarcoma cell line) with zirconia ceramics, whereas gelatin reduced this compatibility. Conclusion: The findings confirm that cold plasma treatment and graft polymerization can promote the immobilization of biomolecules and improve the biocompatibility of zirconia ceramics. This approach can be applied to the modification of zirconia ceramic implants.

Acetic-Acid Plasma-Polymerization on Polymeric Substrates for Biomedical Application.

: Cold plasma is an emerging technology offering many potential applications for regenerative medicine or tissue engineering. This study focused on the characterization of the carboxylic acid functional groups deposited on polymeric substrates using a plasma polymerization process with an acetic acid precursor. The acetic acid precursor contains oxygen and hydrocarbon that, when introduced to a plasma state, forms the polylactide-like film on the substrates. In this study, polymeric substrates were modified by depositing acetic acid plasma film on the surface to improve hydrophilic quality and biocompatibility. The experimental results that of electron spectroscopy for chemical analysis (ESCA) to show for acetic acid film, three peaks corresponding to the C-C group (285.0 eV), C-O group (286.6 eV), and C=O group (288.7 eV) were observed. The resulting of those indicated that appropriate acetic acid plasma treatment could increase the polar components on the surface of substrates to improve the hydrophilicity. In addition, in vitro cell culture studies showed that the embryonic stem (ES) cell adhesion on the acetic acid plasma-treated polymeric substrates is better than the untreated. Such acetic acid film performance makes it become a promising candidate as the surface coating layer on polymeric substrates for biomedical application.

Plasma Deposition and UV Light Induced Surface Grafting Polymerization of NIPAAm on Stainless Steel for Enhancing Corrosion Resistance and Its Drug Delivery Property.

When stainless steel is implanted in human bodies, the corrosion resistance and biocompatibility must be considered. In this study, first, a protective organic silicone film was coated on the surface of stainless steel by a plasma deposition technique with a precursor of hexamethyldisilazane (HMDSZ). Then, ultraviolet (UV) light-induced graft polymerization of N-isopropylacrylamide (NIPAAm) and acrylic acid (AAc) in different molar ratios were applied onto the organic silicone film in order to immobilize thermos-/pH-sensitive composite hydrogels on the surface. The thermo-/pH-sensitive composite hydrogels were tested at pH values of 4, 7.4 and 10 of a phosphate buffer saline (PBS) solution at a fixed temperature of 37 °C to observe the swelling ratio and drug delivery properties of caffeine which served as a drug delivery substance. According to the results of Fourier Transformation Infrared (FTIR) spectra and a potential polarization dynamic test, the silicone thin film formed by plasma deposition not only improved the adhesion ability between the substrate and hydrogels but also exhibited a high corrosion resistance. Furthermore, the composite hydrogels have an excellent release ratio of up to 90% of the absorbed amount after 8h at a pH of 10. In addition, the results of potential polarization dynamic tests showed that the corrosion resistance of stainless steel could be improved by the HMDSZ plasma deposition.

An Effective Cell Coculture Platform Based on the Electrospun Microtube Array Membrane for Nerve Regeneration.

Recently, a novel substrate known as an electrospun polylactic acid (PLLA) microtube array membrane (MTAM) was successfully developed as a cell coculture platform. Structurally, this substrate is made up of one-to-one connected, ultrathin, submicron scale fibers that are arranged in an arrayed formation. Its unique structure confers several key advantages which are beneficial in a cell coculture system. In this study, the interaction between rat fetal neural stem cells (NSC) and astrocytes was examined by comparing the outcome of a typical Transwell-based coculture system and that of an electrospun PLLA MTAM-based coculture system. Compared to tissue culture polystyrene (TCP) and Transwell coculture inserts, a superior cell viability of NSC was observed when cultured in lumens of electrospun PLLA MTAM (with supportive immunostaining images). Reverse transcription polymerase chain reaction revealed a strong interaction between astrocytes and NSC through a higher expression of doublecortin and a lower expression of nestin. These data demonstrate that MTAM is clearly a better coculture platform than the traditional Transwell system.

In situ synthesis of high swell ratio polyacrylic acid/silver nanocomposite hydrogels and their antimicrobial properties.

Silver nanocomposites embedded within a polymer matrix have attracted attention in recent years. Ionic polymer hydrogels comprise networks of chemically or physically cross-linked polymers that swell considerably in an appropriate solvent. In this study, we used a solution of the carboxylic monomer acrylic acid and silver nitrate to prepare nanocomposite hydrogels through ultraviolet (UV)-light irradiation. Silver-impregnated biomaterial composed of acrylic acid contains only a monomer and no cross-linker. The formation of hydrogels and reduction of silver nanoparticles were affected by the preparation parameters, that is, the monomer concentration and silver nitrate concentration. The morphology, structure, and size of the silver nanocomposite hydrogels were evaluated through field emission scanning electron microscopy and UV-visible absorption. The antimicrobial activity of the samples was tested against fourstandard strains Candida albicans, Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli; and five clinical bacterial isolates Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Acinetobacter baumannii, Klebsiella pneumonia. The silver nanocomposite hydrogels exhibited an interconnected porous structure and could absorb 400 to 550g of deionized water per gram of dried hydrogel. Moreover, these hydrogels produced a strong antibacterial effect, which can be useful in developing new superabsorbent antimicrobial pharmaceutical products.

Cell proliferation on PVA/sodium alginate and PVA/poly(γ-glutamic acid) electrospun fiber.

To overcome the obstacles of easy dissolution of PVA nanofibers without crosslinking treatment and the poor electrospinnability of the PVA cross-linked nanofibers via electrospinning process, the PVA based electrospun hydrogel nanofibers are prepared with post-crosslinking method. To expect the electrospun hydrogel fibers might be a promising scaffold for cell culture and tissue engineering applications, the evaluation of cell proliferation on the post-crosslinking electrospun fibers is conducted in this study. At beginning, poly(vinyl alcohol) (PVA), PVA/sodium alginate (PVASA) and PVA/poly(γ-glutamic acid) (PVAPGA) electrospun fibers were prepared by electrospinning method. The electrospun PVA, PVASA and PVAPGA nanofibers were treated with post-cross-linking method with glutaraldehyde (Glu) as crosslinking agent. These electrospun fibers were characterized with thermogravimetry analysis (TGA) and their morphologies were observed with a scanning electron microscope (SEM). To support the evaluation and explanation of cell growth on the fiber, the study of 3T3 mouse fibroblast cell growth on the surface of pure PVA, SA, and PGA thin films is conducted. The proliferation of 3T3 on the electrospun fiber surface of PVA, PVASA, and PVAPGA was evaluated by seeding 3T3 fibroblast cells on these crosslinked electrospun fibers. The cell viability on electrospun fibers was conducted with water-soluble tetrazolium salt-1 assay (Cell Proliferation Reagent WST-1). The morphology of the cells on the fibers was also observed with SEM. The results of WST-1 assay revealed that 3T3 cells cultured on different electrospun fibers had similar viability, and the cell viability increased with time for all electrospun fibers. From the morphology of the cells on electrospun fibers, it is found that 3T3 cells attached on all electrospun fiber after 1day seeded. Cell-cell communication was noticed on day 3 for all electrospun fibers. Extracellular matrix (ECM) productions were found and cell-ECM adhesion was shown on day 7. The cell number was also increased on all of the crosslinked electrospun fibers. It seems that the PVA based electrospun hydrogel nanofibers prepared with post-crosslinking method can be used as scaffold for tissue engineering.

Biopolymer coated gold nanocrystals prepared using the green chemistry approach and their shape-dependent catalytic and surface-enhanced Raman scattering properties.

This study deals with the preparation of multi-shaped nanoscale gold crystals under synthetically simple, green, and efficient conditions using a seed-mediated growth approach in the presence of hyaluronic acid (HA). These highly biocompatible multi-shaped gold nanocrystals were examined to evaluate their catalytic and surface enhanced Raman scattering (SERS) properties. The results show that the size and shape of the nanocrystals are mainly correlated to the amount of seed, seed size, HA concentration, and reaction temperature. Gold seeds accelerate the reduction of the gold precursor to form gold nanocrystals using HA. The HA serves as a reducing agent and a growth template for the reduction of Au(III) and nanocrystal stabilization. The multi-shaped gold nanocrystals showed superior catalytic properties and higher SERS performance. The simple, green approach efficiently controls the nanocrystals and creates many opportunities for future applications.

New nerve regeneration strategy combining laminin-coated chitosan conduits and stem cell therapy.

Nerve regeneration remains a difficult challenge due to the lack of safe and efficient matrix support. We designed a laminin (LN)-modified chitosan multi-walled nerve conduit combined with bone marrow stem cell (BMSC) grating to bridge a 10 mm long gap in the sciatic nerve of Sprague-Dawley rats. The repair outcome was monitored during 16 weeks after surgery. Successful grafting of LN onto the chitosan film, confirmed by immunolocalization, significantly improved cell adhesion. In vivo study showed that newly formed nerve cells covered the interior of the conduit to connect the nerve gap successfully in all groups. The rats implanted with the conduit combined with BMSCs showed the best results, in terms of nerve regrowth, muscle mass of gastrocnemius, function recovery and tract tracing. Neuroanatomical horseradish peroxidase tracer analysis of motor neurons in the lumbar spinal cord indicated that the amount and signal intensity were significantly improved. Furthermore, BMSCs suppressed neuronal cell death and promoted regeneration by suppressing the inflammatory and fibrotic response induced by chitosan after long-term implantation. In summary, this study suggests that LN-modified chitosan multi-walled nerve conduit combined with BMSCs is an efficient and safe conduit matrix for nerve regeneration.

Post treatment of plasma-polymerized SnOx organic-like films with poly ethylene glycol for improving CO gas sensitivity.

In this study, a new room temperature type gas sensor device based on plasma deposition of tetramethyltin (TMT) and O2 organically hybridized film followed by post treatment on the deposited film was developed for improving CO gas sensitivity and distinguishing from methane, butane, and carbon monoxide gases in the test environment. Plasma deposited SnOx thin film was first produced from TMT and O2 gas mixtures at room temperature, and then post treatments on the SnOx thin films were carried out by either spin coating with poly ethylene glycol (PEG) or surface grafting with p-styrenesulfonic acid sodium salt (Nass). It was found that the gas sensor spin coating post treated with PEG exhibits linear response to CO gas with the sensitivity not affected by methane and butane gases. For CO concentrations ranging from 30 to 650 ppm, steep change in the sensor resistance can be detected without warming up the sensor.

Bostrycin, a novel coupling agent for protein immobilization and prevention of biomaterial-centered infection produced by Nigrospora sp. No. 407.

Bostrycin, a red antibacterial agent with tetrahydroanthraquinone structure, has been isolated from Nigrospora sp. No. 407. This study investigated the potential antibacterial and multifunctional properties of matrixes through immobilization of bostrycin on their surface for immobilization of protein and prevention of bacterial growth. Bostrycin was immobilized on nonwoven polypropylene (PP) fabric by a technique using glutaraldehyde and polyethyleneimine for the activation of the surface. Glucose oxidase immobilized on bostrycin-treated nonwoven PP fabric showed high activity. The immobilization process improved thermal stability of the enzymes. During repeated assay for 30 cycles, the enzyme activity dropped to only 70% of the initial activity. Both bostrycin-treated nonwoven PP fabric sample and subsequently immobilized glucose oxidase sample on the surface also still exhibited a bacteriostatic effect. This is the first study to show that bostrycin is a promising coupling agent for surface modification on matrix and its potential applications in protein immobilization and biomaterial-centered infection.

Detecting cells on the surface of a silver electrode quartz crystal microbalance using plasma treatment and graft polymerization.

This paper utilizes a silver electrode quartz crystal microbalance (QCM) mass sensor to detect the physiology of cells. This study also investigates the plasma surface modification of silver electrode QCMs through deposition of hexamethyldisilazane (HMDSZ) films as a protection film. To improve the cell growth, this paper also performs post-treatments by surface-grafting acrylic acid (AAc), acrylamide (AAm), and oxygen plasma treatment onto the QCM electrodes. Experimental results indicate that plasma deposition is a useful technique to protect the surface of silver electrodes. This technique extends the unpeeling time of silver electrodes from 1 to 7 days. The hydrophilic silver electrode QCM surface modified by AAm exhibited a better storage time effect than other post-treatments.

Immobilization of Chinese herbal medicine onto the surface-modified calcium hydrogenphosphate.

To accelerate the healing of bone defects or for healing to take place, it is often necessary to fill them with suitable substance. Various artificial materials defects have been developed. Among these, calcium phosphates and bioactive glass have been proven to be biocompatibile and bioactive materials that can chemically bond with bone, and have been successfully used clinically for repair of bone defects and augmentation of osseous tissue. However, those bioceramics have only the property of osteoconduction without any osteoinduction. Many ligands have been physicochemically absorbed onto substrates to enhance cell-substrate interactions. Although widely developed, they are still limited to use in long-term implantation because of their half-life period. Thus, some interfacial modification will be required for enhancing the efficacy of the delivery system. These models involve the immobilization of biologically active ligands of natural and synthetic origin onto various substrates to produce an interface with stronger chemical bond between ligand and substrate. The advantage of covalently immobilizing a ligand is that a chemical bond is present to prevent ligand or medicine from desorption. In our study, a two-step chemical immobilization was performed to surface-modified calcium hydrogenphosphate powders. The first was to modify the surface of calcium hydrogen-phosphate (CHP) with a coupling agent of hexanmethylene diisocyanate (HMDI). CHP surface modified by HMDI is abbreviated as MCHP. The linkage between CHP and HMDI will be characterized by FTIR. The second step was to immobilize chemically Gusuibu onto MCHP. Moreover, the sorption and desorption of Gusuibu was evaluated and quantitatively analyzed by spectrophotometer and HPLC. Bioceramic CHP was surface-modified by a two-step chemical immobilization. First, the surface of calcium hydrogen-phosphate (CHP) was successfully modified with coupling agent of hexanmethylene diisocyanate (HMDI). The first step was also activated the surface of CHP to induce primary amine terminator. The reaction of this functional group with Gusuibu was the second step. We confirmed simultaneously that Gusuibu could be immobilized chemically onto the surface of MCHP. Although some immobilized Gusuibu was also released rapidly at the first 12h, the degree of the released Gusuibu was lower than both by Gusuibu-adsorbing MCHP and Gusuibu-adsorbing CHP.