Mineralized collagen coatings formed by electrochemical deposition.

Understanding and controlling the process of electrochemical deposition (ECD) of a mineralized collagen coating on metallic orthopedic implants is important for engineering highly bioactive coatings. In this work, the influence of different ECD parameters was investigated. The results showed that the mineralization degree of the coatings increased with deposition time, voltage potential and H2O2 addition, while chitosan addition led to weakening of mineralization, heavy mineralization resulted in a porous coating morphology. Furthermore, two typical coatings, dense and porous, were analyzed to investigate their microstructure and evaluated for their cytocompatibility; the dense coating showed better osteoblast adhesion and proliferation. Based on our understanding of how the different coating parameters influenced the coating, we proposed an ECD process in which the pH gradient near the cathode and the collagen isoelectric point were suggested to play crucial roles in controlling the mineralization and morphology of the coatings. The proposed ECD process may offer a guide for controlled deposition of a desired bioactive coating.

Fabrication and evaluation of Zn containing fluoridated hydroxyapatite layer with Zn release ability.

A biphasic layer with a Zn-containing beta-tricalcium phosphate (ZnTCP) phase and a fluoridated hydroxyapatite (FHA) phase on titanium alloy substrate was prepared by the sol-gel technique. Scanning electron microscopy and energy-dispersive X-ray analysis results showed the ZnTCP/FHA layer to have a heterogeneous surface with microscaled gibbous structures originating from ZnTCP particle agglomeration. This layer had a slow and sustained Zn release behavior. The scratch test result of the ZnTCP/FHA layer was 489+/-4mN, indicating good interface bonding between the layer and substrate. The ZnTCP/FHA layer supported cell growth, and showed a statistically significant increase in cell viability in comparison with another biphasic layer (TCP/FHA) without Zn. This work demonstrates that the present biphasic ZnTCP/FHA layer has the potential to play a significant role in enhancing bone growth when used as the outermost part of bioactive coatings on metallic implants.

Preparation, characterization and cytocompatibility of porous ACP/PLLA composites.

The purpose of this work was to incorporate amorphous calcium phosphate (ACP) into porous poly(L-lactic acid) (PLLA), because ACP is capable of fast phase transformation and morphological change in body fluid, such, a desired pore wall surface within bone tissue engineering scaffolds can be created. A highly porous ACP/PLLA composite was prepared by a thermally induced phase separation technique. The results showed that the composite had an interconnected pore structure with 100 mum macropores and 10 mum micropores, and 91% porosity; 40 nm primary particles of ACP were agglomerated to 3 mum aggregates, and the aggregates were homogeneously distributed in pore walls; These aggregates showed to be in situ transformed into bone-like apatite after 1 h soaking in phosphate buffered saline solution. Human osteoblast-like cell culture showed that the ACP/PLLA composite had better cell adhesion and alkaline phosphotase activity than pure PLLA. This study demonstrates that the ACP/PLLA composite can enhance cytocompatibility and could act as a promising scaffold for bone tissue engineering.

Preparation and characterization of porous beta-tricalcium phosphate/collagen composites with an integrated structure.

Porous beta-tricalcium phosphate (TCP)/collagen composites with different beta-TCP/collagen weight ratio were prepared. The influences of the preparation conditions on the microstructure of porous composite and the joint status of beta-TCP particles with collagen fibrils were characterized by X-ray diffractometer, scanning electron microscopy and transmission electron microscopy. The results showed: (1) an acid treatment could effectively disassemble collagen fibrils; (2) in the resulting porous composites, beta-TCP particles homogenously existed on the skeleton of the collagen fibril network and bonded tightly to both the fibrils and themselves. The tight bonding formation could be due to the reaction between Ca ions in the particles and carboxyl groups in collagen polypeptide chains and due to the reprecipitation of partially dissolved beta-TCP during synthesis. The tight bonding between beta-TCP particles and collagen fibrils in the composites demonstrated an integrated structure, which was reproducible when beta-TCP/collagen ratio ranged from 2 to 4. Such integrated structure would make significant contributions in reliably tailoring properties of the porous composites by varying beta-TCP content. In addition, the porous composites had large porosity (approximately 95%) and appropriate pore size (approximately 100 microm), showed no negative impact in cytotoxicity assay and complete bone tissue regeneration after 12 weeks in animal test.

Sol-gel preparation and in vitro test of fluorapatite/hydroxyapatite films.

Fluorapatite/hydroxyapatite (FA/HA) films have been demonstrated to be a good alternative to pure hydroxyapatite (HA) ones in medical applications because of their bioactivity and relatively low solubility. In this study, Ca(NO(3))(2), P(2)O(5), and HPF(6) were used to prepare FA/HA films on Ti6Al4V substrate with the use of a sol-gel method. The F contents in the films could be tailored by adjusting the amount of HPF(6) added. The in vitro evaluation of the films was carried out in both SBF9# solution and TRIS buffer solution. The films with appropriate F contents showed a better ability to induce calcium phosphate deposition on their surfaces than either pure HA film and FA/HA films with even higher F content, as well as smaller dissolution amounts than HA film in TRIS buffer solution. Hence, the FA/HA films obtained in this work integrate both good bioactivity and stability, and could be a better choice for bioactive film on titanium alloys to produce high-quality implants.

Polycrystalline ZnS(x)Se(1 - x) thin films deposited on ITO glass by MBE.

MBE growth of ZnS(x)Se(1 - x) thin films on ITO coated glass substrates were carried out using ZnS and Se sources with the substrate temperature ranging from 270 degrees C to 330 degrees C . The XRD theta/2theta spectra resulted from these films indicated that the as-grown polycrystalline ZnS(x)Se(1 - x) thin films had a preferred orientation along the (111) planes. The evaluated crystal sizes as deduced from the FWHM of the XRD layer peaks showed strong growth temperature dependence, with the optimized temperature being about 290 degrees C. Both AFM and TEM measurements of these thin films also indicated a similar growth temperature dependence. High quality ZnS(x)Se(1 - x) thin film grown at the optimized temperature had the smoothest surface with lowest RMS value of 1.2 nm and TEM cross-sectional micrograph showing a well defined columnar structure.

Improvement of drug elution in thin mineralized collagen coatings with PLGA-PEG-PLGA micelles.

A mineralized collagen (MC) coating on metallic implants has shown great potential as orthopedic material due to high biological responses. However, their drug delivery capacity remains unsatisfactory since a serious burst release may occur and long-term release is hard to be achieved. Aiming to improve the drug-eluting capability, we incorporated drug-loaded PLGA-PEG-PLGA (PPP) micelles into the thin coating. The in vitro release profiles showed that the burst release in the initial 8 h of the modified coating decreased from 81% to 58% compared to MC coating alone; meanwhile, the release duration was prolonged from 3 days to 1 week. Additionally, the release kinetics of vancomycin hydrochloride (VH, the model drug) could be adjusted by changing the size and concentration of PPP micelles. Interestingly, less initial release of VH caused by micelle immobilization did not affect the antibacterial activity in the early stage of implantation according to the antimicrobial test. The cytocompatibility assay demonstrated that the VH-loaded PPP micelles did not have negative effect on the bioactivity of coating which greatly enhanced cell activity compared to bare Ti substrates. Thus, the MC coatings with PPP micelles could be an effective implant route for bone repair.

Incorporation of chitosan nanospheres into thin mineralized collagen coatings for improving the antibacterial effect.

It is desired that the coatings on metallic implants have both excellent biological responses and good loading-release capacities of biological factors or drugs. So far, the challenge still remains, because the morphology and composition of the bioactive coatings are usually not favorable for accommodating drug molecules. In this study, we adopted an approach of incorporating chitosan nanospheres into a thin mineralized collagen coating; this approach is based on the good loading-release behavior of the nanospheres and the good cytocompatibility of the thin coating. The incorporation of chitosan nanospheres into the mineralized collagen coatings was realized by electrolytic co-deposition. The morphologies and microstructures of the resulting coatings were characterized by SEM, and the phase and chemical compositions of the coatings were measured by XRD and FTIR. The loading-release capacity for vancomycin hydrochloride (VH) was determined by ultraviolet spectrophotometry. MTS assay was used to evaluate cytocompatibility, and in vitro bacterial adhesion was tested for assessing the antibacterial effects of the VH-loaded coatings. The chitosan nanospheres adhered tightly to collagen fibrils. The incorporated coatings facilitated the sustained release of VH, and had a clear antibacterial effect. The incorporation of chitosan nanospheres into mineralized collagen coatings demonstrates an effective way to improve the drug loading-release capacity for the thin coatings. This formulation had a highly effective biological response.

Effect of PEG amount in amorphous calcium phosphate on its crystallized products.

Biphasic alpha-tricalcium phosphate/beta-tricalcium phosphate (alpha/beta-TCP) with a designed phase ratio is thought to have controllable dissolution-reprecipitation behavior that is significant in the repair and regeneration of bone. Amorphous calcium phosphate (ACP) was selected as a precursor to prepare biphasic alpha/beta-TCP. The influence of polyethylene glycol (PEG) content in ACP on its crystallization, or on the phase ratio of the resulting biphasic TCP, was investigated. ACP was synthesized by the reaction of Ca(NO(3))(2) with (NH(4))(2)HPO(4) using PEG as an additive. Depending on the amount of PEG addition, resulting ACP could be crystallized to alpha-TCP, beta-TCP or biphasic alpha/beta-TCP after heat-treatment at 800 degrees C, showing that PEG addition is a critical factor to tailor the phase ratio of biphasic alpha/beta-TCP. One reason for the influence of PEG is that ACP with different PEG content could have two types of unit structures that tend to form alpha-TCP and beta-TCP after crystallization.

Electrolytic deposition of octacalcium phosphate/collagen composite coating on titanium alloy.

Osteointegration of titanium or its alloy with bone can be greatly improved by calcium phosphate coatings, and further enhanced by an extracellular matrix protein layer such as collagen. In this study, an octacalcium phosphate (OCP)/collagen composite coating layer on Ti6Al4V substrate was prepared using electrolytic deposition method. A layer of OCP mineral consisting of flake-like crystals was first formed on the Ti6Al4V substrate. Subsequently, mineralized collagen fibrils were deposited on the former OCP layer. These collagen fibrils were interconnected and well adhered on the OCP layer so that they were immobilized. The microstructure of the composite coating varied with collagen concentration in the electrolyte. This study could offer a possibility of fabricating a desired surface matrix on orthopedic implants to enhance bone formation and fixation of implants.

Porous beta-tricalcium phosphate/collagen composites prepared in an alkaline condition.

Bone substitute materials with natural bone-like structure are considered to be favorable for bone regeneration. In this work, porous beta-tricalcium phosphate (beta-TCP)/collagen composite consisting of bone-like microstructural units was prepared using nanosized beta-TCP particles and alkaline-disassembled collagen. The resulting composite showed a good interconnecting porous structure with approximately 90% porosity and 100 approximately 300 microm pore size. The pore walls were dense, and the combination status of collagen and nanosized beta-TCP particles demonstrated that nanosized beta-TCP particles tightly connected collagen microfibrils as a bone-like microstructural unit. MTT and alkaline phosphatase (ALP) assays showed that the porous composite had enhanced effects on cellular proliferation and activity of osteoblast compared with a control of pure collagen. It is suggested that the adoption of nanosized beta-TCP particles is a main contribution to the formation of the composite with a bone-like microstructural unit, and the unique microstructure could be a main role for the composite to have the positive influence on osteoblast cell proliferation.

In vitro bioactivity and osteoblast-like cell test of zinc containing fluoridated hydroxyapatite films.

Zinc containing fluoridated hydroxyapatite (ZnFHA) films on Ti6Al4V substrates was prepared using sol-gel dip-coating method. The release of zinc ions from ZnFHA film was controlled mainly by the zinc content in the film. The release behavior showed an initial rapid increase release followed by a tapering-off and directed to a constant value at longer time. After soaking in SBF for 8 days, a layer was deposited and completely covered the original surface of the ZnFHA film, indicating good in vitro "bioactivity." The osteoblast-like MG63 cells were seeded on the ZnFHA films; FHA film and Ti6Al4V substrate were used as control. The cell culture result showed that cell adhesion and proliferation on ZnFHA films were significantly increased compared with the controls. The results in this work suggest that ZnFHA films on Ti6Al4V substrates can function as an implant with good bioactivity and cytocompatibility.