Xuanyong Liu

Antibacterial activity and increased bone marrow stem cell functions of Zn-incorporated TiO2 coatings on titanium

In this work, zinc was incorporated into TiO2 coatings on titanium by plasma electrolytic oxidation to obtain the implant with good bacterial inhibition ability and bone-formability. The porous and nanostructured Zn-incorporated TiO2 coatings are built up from pores smaller than 5 μm and grains 20-100 nm in size, in which the element Zn exists as ZnO. The results obtained from the antibacterial studies suggest that the Zn-incorporated TiO2 coatings can greatly inhibit the growth of both Staphylococcus aureus and Escherichia coli, and the ability to inhibit bacteria can be improved by increasing the Zn content in the coatings. Moreover, the in vitro cytocompatibility evaluation demonstrates that the adhesion, proliferation and differentiation of rat bone marrow stem cells (bMSC) on Zn-incorporated coatings are significantly enhanced compared with Zn-free coating and commercially pure Ti plate, and no cytotoxicity appeared on any of the Zn-incorporated TiO2 coatings. Moreover, bMSC express higher level of alkaline phosphatase activity on Zn-incorporated TiO2 coatings and are induced to differentiate into osteoblast cells. The better antibacterial activity, cytocompatibility and the capability to promote bMSC osteogenic differentiation of Zn-incorporated TiO2 coatings may be attributed to the fact that Zn ions can be slowly and constantly released from the coatings. In conclusion, innovative Zn-incorporated TiO2 coatings on titanium with excellent antibacterial activity and biocompatibility are promising candidates for orthopedic and dental implants.

Apatite formed on the surface of plasma-sprayed wollastonite coating immersed in simulated body fluid.

Wollastonite coatings on titanium alloys substrates were prepared by plasma spraying and incubated in simulated body fluids for different periods to investigate the nucleation and growth of apatite on their surface. Surface structural changes of the specimens were analyzed by XRD and IR technologies. SEM and EDS were used to observe surface morphologies and determine the composition of wollastonite coatings before and after immersion in simulated body fluid. The changes in the concentrations of calcium, silicon and phosphorus in the simulated body fluids due to the immersion of the specimens were measured by inductively coupled plasma atomic emission spectroscopy. The results obtained showed that hydroxycarbonate apatite can be formed on the surface of the coating soaked in SBF for 1 day. With longer immersion periods, the coating surface was covered by hydroxycarbonate apatite, which indicated that the wollastonite coating possesses good bioactivity.

Biological actions of silver nanoparticles embedded in titanium controlled by micro-galvanic effects.

Titanium embedded with silver nanoparticles (Ag NPs) using a single step silver plasma immersion ion implantation (Ag-PIII) demonstrate micro-galvanic effects that give rise to both controlled antibacterial activity and excellent compatibility with osteoblasts. Scanning electron microscopy (SEM) shows that nanoparticles with average sizes of about 5 nm and 8 nm are formed homogeneously on the titanium surface after undergoing Ag-PIII for 0.5 h and 1 h, respectively. Transmission electron microscopy (TEM) and X-ray photoelectron spectroscopy (XPS) indicate that those nanoparticles are metallic silver produced on and underneath the titanium surface via a local nucleation process from the solid solution of α-Ti(Ag). The Ag-PIII samples inhibit the growth of both Staphylococcus aureus and Escherichia coli while enhancing proliferation of the osteoblast-like cell line MG63. Electrochemical polarization and Zeta potential measurements demonstrate that the low surface toxicity and good cytocompatibility are related to the micro-galvanic effect between the Ag NPs and titanium matrix. Our results show that the physico-chemical properties of the Ag NPs are important in the control of the cytotoxicity and this study opens a new window for the design of nanostructured surfaces on which the biological actions of the Ag NPs can be accurately tailored.

The synergistic effect of hierarchical micro/nano-topography and bioactive ions for enhanced osseointegration

Both surface chemistry and topography have significant influence on good and fast osseointegration of biomedical implants; the main goals in orthopeadic, dental and maxillofacial surgeries. A surface modification strategy encompassing the use of bioactive trace elements together with surface micron/nano-topographical modifications was employed in this study in an attempt to enhance the osseointegration of Ti alloy (Ti-6Al-4V), a commonly used implant. Briefly, we developed strontium-substituted hardystonite (Sr-HT) ceramic coating with a hierarchical topography where the nanosized grains were superimposed in the micron-rough coating structure. Its ability to induce new bone formation was evaluated by an in vivo animal model (beagle dogs). Hardystonite (HT), classic hydroxyapatite (HAp) coated and uncoated Ti-alloy implants were parallelly investigated for comparison. In addition, we investigated the effects of surface topography and the dissolution products from the coatings on the in vitro bioactivity using canine bone marrow mesenchymal stem cells (BMMSCs) cultured on the implant surface as well as using extracts of the coated implants. Micro-CT evaluation, histological observations, biomechanical test (push-out test) and sequential fluorescent labeling and histomorphometrical analysis consistently demonstrated that our developed Sr-HT-coated Ti-alloy implants have the highest osseointegration, while the uncoated implants had the lowest. The osseointegration ability of HAp-coated Ti alloy was inferior to that seen for HT- and Sr-HT-coated Ti alloy. We demonstrated that the dissolution products, particularly strontium (Sr) from the Sr-HT-coated implants, enhanced the ALP activity and in vitro mineralization ability, while the micro/nano-topography was more related to the promotion of cell adhesion. Those results suggest that our developed Sr-HT coatings have the potential for future use as coatings for orthopedic/dental and maxillofacial devices.

Antibacterial activity of large-area monolayer graphene film manipulated by charge transfer

Graphene has attracted increasing attention for potential applications in biotechnology due to its excellent electronic property and biocompatibility. Here we use both Gram-positive Staphylococcus aureus (S. aureus) and Gram-negative Escherichia coli (E. coli) to investigate the antibacterial actions of large-area monolayer graphene film on conductor Cu, semiconductor Ge and insulator SiO2. The results show that the graphene films on Cu and Ge can surprisingly inhibit the growth of both bacteria, especially the former. However, the proliferation of both bacteria cannot be significantly restricted by the graphene film on SiO2. The morphology of S. aureus and E. coli on graphene films further confirms that the direct contact of both bacteria with graphene on Cu and Ge can cause membrane damage and destroy membrane integrity, while no evident membrane destruction is induced by graphene on SiO2. From the viewpoint of charge transfer, a plausible mechanism is proposed here to explain this phenomenon. This study may provide new insights for the better understanding of antibacterial actions of graphene film and for the better designing of graphene-based antibiotics or other biomedical applications.

Osteogenic activity and antibacterial effects on titanium surfaces modified with Zn-incorporated nanotube arrays

Titanium implants having enhanced osteogenic activity and antibacterial property are highly desirable for the prevention of implant associated infection and promotion of osseointegration. In this study, coatings containing titania nanotubes (NTs) incorporated with zinc (NT-Zn) are produced on Ti implants by anodization and hydrothermal treatment in Zn containing solutions. The amount of incorporated Zn can be adjusted by varying the structural parameters such as the nanotube diameter and length as well as hydrothermal treatment time. The suitable NT-Zn coatings with good intrinsic antibacterial properties can prevent post-operation infection. Excellent osteogenesis inducing ability in the absence of extraneous osteogenic supplements is demonstrated and the ERK1/2 signaling is found to be involved. The NT-Zn structure which is simple, stable, and easy to produce and scale up has immense potential in bone implant applications.

Bioactivity of plasma sprayed dicalcium silicate coatings

Dicalcium silicate coatings on titanium alloys substrates were prepared by plasma spraying and immersed in simulated body fluids for a period of time to investigate the nucleation and growth of apatite on the surface of the coatings. Surface structural changes of the specimens were analyzed by XRD and IR technologies. SEM and EDS were used to observe surface morphologies and determine the composition of dicalcium silicate coatings before and after immersion in simulated body fluid. The plasma sprayed dicalcium silicate coating was bonding tightly to the substrate. The coating was mainly composed of beta-Ca2SiO4 and glassy phase. A dense carbonate-containing hydroxyapatite (CHA) layer was formed on the surface of the plasma sprayed dicalcium silicate coating soaked in SBF solution for 2 days. In addition, a silica-rich layer was also observed between CHA layer and coatings. With an increase in the immersion time, the CHA layer gradually became thicker. The results obtained indicated that the plasma sprayed dicalcium silicate coating possesses excellent bioactivity.

In vivo evaluation of plasma sprayed hydroxyapatite coatings having different crystallinity.

In this paper, hydroxyapatite (HA) coatings having the crystallinities of 56% and 98% were deposited by the plasma spraying and vapor-flame treatment process. The phase composition and crystallinity of the coatings were investigated by X-ray diffraction and infrared spectra. The dissolution behavior of the coatings in tris-buffer solutions was examined. The results obtained indicated that the coating having the high crystallinity showed the lower dissolution as compared to the low crystallinity coating. The bone bonding ability of HA coatings were observed in vivo by implanted in dogs femur. After 3 months implantation, the high crystallinity coating showed the higher shear strengths and remained integrated, whereas the separation of the coating fragments was clearly observed in the coating having low crystallinity.

Influences of ionic dissolution products of dicalcium silicate coating on osteoblastic proliferation, differentiation and gene expression

This work aims to explore the influence of the ionic products of dicalcium silicate coating on osteoblastic proliferation and differentiation, as well as on the expression of BMP2 and its signal transducers Smad1, 6 and 7 in MG-63 osteoblast-like cells. Plasma-sprayed dicalcium silicate coatings were soaked in DMEM to obtain culture media containing the ionic dissolution products of dicalcium silicate coating (Ca2SiO4-DMEM). MG63 osteoblast-like cells were cultured in Ca2SiO4-DMEM (experimental group) for 3-12 days, while those cultured in normal DMEM served as control (control group). MTT assay was used to evaluate cell viability and proliferation. Alkaline phosphatase activity (ALP), osteocalcin (OC) and type I collagen (COLI) were investigated as differentiation markers. Gene expression of BMP2 and Smad1, 6, 7 was also detected. BMP2 protein was examined by ELISA assay. Alizarin Red-S (AR-S) assay was used to detect mineralization. The results demonstrated that Si concentration in Ca2SiO4-DMEM is markedly higher than that in normal DMEM. Compared to the control group, MG63 cells of the experimental group exhibited upregulated proliferation on day 3, and markedly upregulated gene expression of the differentiation markers, especially on days 9 and 12 for OC and on days 3, 6 and 9 for ALP. Gene expression of BMP2 and Smad1, as well as BMP2 protein secreted into culture media, was also upregulated in the experimental group, while gene expression of Smad6 and 7 was not influenced. AR-S assay indicated a higher calcium mineral content deposition in cells of the experimental group. In conclusion, the ionic products of plasma-sprayed dicalcium silicate coating are beneficial to the proliferation and differentiation of MG63 osteoblast-like cells.

Plasma sprayed wollastonite/TiO2 composite coatings on titanium alloys.

Wollastonite/TiO2 composite coatings were prepared using plasma spraying technology onto Ti-6Al-4V substrate. The composite coatings exhibit obvious lamellar structure with alternating wollastonite coating and TiO2 coating. No obvious cracks exist on the interface between coatings and substrate. In the case of composite coatings, the primarily crystalline phases of the coatings are wollastonite and rutile, indicating wollastonite and TiO2 did not react during plasma spraying process. Some of rutile in the powders transforms into anatase due to plasma spraying. The mean bond strength of the composite coatings is higher than 30 MPa. The Vickers microhardness of coatings increase with the increase in the content of TiO2. Wollastonite/TiO2 composite coatings were soaked in simulated body fluid to examine their bioactivity. Carbonate-containing hydroxyapatite (CHA) layer was formed on the surface of the wollastonite and W7T3 coatings soaked in simulated body fluid, while was not formed on the surface of the TiO2 and W3T7 coatings after immersion. In addition, a rich-silica layer appeared at the interface of CHA and wollastonite and W7T3 coatings. In order to investigate the cytocompatibility of the coatings, osteoblast was seeded onto the surface of the coatings. The scanning electron microscopy observation showed that the addition of wollastonite promote the proliferation of osteoblast. It is enough to prove that the wollastonite and wollastonite/TiO2 composite coatings possess excellent cytocompatibility.