Built-up sodium alginate/chlorhexidine multilayer coating on dental implants with initiating anti-infection and cyto-compatibility sequentially for soft-tissue sealing.

Soft-tissue sealing at transmucosal sites is very important for preventing the invasion of pathogens and maintaining the long-term stability and function of dental implants. However, the colonization of oral pathogens on the implant surface and surrounding soft tissues can disturb the early establishment of soft-tissue sealing and even induce peri-implant infection. The purpose of this study was to construct two antibacterial coatings with 5 or 10 sodium alginate/chlorhexidine bilayers on titanium surfaces using layer-by-layer self-assembly technology to promote soft-tissue sealing. The corresponding chemical composition, surface topography, wettability and release behaviour were investigated to prove that the resultant coating of sodium alginate and chlorhexidine was coated on the porous titanium surface. In-vitro and in-vivo antibacterial results showed that both prepared coatings inhibited or killed the bacteria on their surfaces and the surrounding areas to prevent plaque biofilm formation, especially the coating with 10 bilayers. Although both coatings inhibited the initial adhesion of fibroblasts, the cytocompatibility gradually improved with coating degradation. More importantly, both coatings achieved cell adhesion and proliferation in an in-vitro bacterial environment and effectively alleviated bacteria-induced subcutaneous inflammation in-vivo. Therefore, this study demonstrated that the multilayered coating could prevent implant-related infections in the initial stage of implant surgery and then improve soft-tissue integration with implant devices.

A programmed surface on dental implants sequentially initiates bacteriostasis and osseointegration.

Osteogenesis surrounding dental implants is initiated by a series of early physiological events, including the inflammatory response. However, the persistence of an anti-infection surface often results in compromised histocompatibility and osseointegration. Here, we presented a programmed surface containing both silver nanoparticles (AgNPs) and silver ions (Ag+) with a heterogeneous structure and time-dependent functionalities. The AgNPs were located at the surface of the heparin-chitosan polyelectrolyte coating (PEM), whereas Ag+ was distributed at both the surface and inside of the coating under optimized conditions (pH=4). The optimized coating (Ag-4) exhibited potent bactericidal activity at the early stage (12 and 24 h after inoculation) and a sustained antibacterial efficacy in the subsequent stage (one or two weeks), as it gradually depleted. Furthermore, compared to coatings with sustained high silver concentrations in bacteria-cell coculture experiments, the degradable Ag-4 coating demonstrated improved cytocompatibility, better cell viability, and morphology over time. At a later stage (within one month), the in vivo test revealed that Ag-4-coated titanium had superior histocompatibility and osteogenesis outcomes compared to bare titanium in a bacteria-exposed environment. The programmed surface of dental implants presented in this study offers innovative ideas for sequential antibacterial effects and osseointegration.

A homogeneous dopamine-silver nanocomposite coating: striking a balance between the antibacterial ability and cytocompatibility of dental implants.

Silver has been widely used for surface modification to prevent implant-associated infections. However, the inherent cytotoxicity of silver greatly limited the scope of its clinical applications. The construction of surfaces with both good antibacterial properties and favorable cytocompatibility still remains a challenge. In this study, a structurally homogeneous dopamine-silver (DA/Ag) nanocomposite was fabricated on the implant surface to balance the antibacterial activity and cytocompatibility of the implant. The results show that the DA/Ag nanocomposites prepared under the acidic conditions (pH = 4) on the titanium surface are homogeneous with higher Ag+ content, while an obvious core (AgNPs)-shell (PDA) structure is formed under neutral (pH = 7) and alkaline conditions (pH = 10), and the subsequent heat treatment enhanced the stability of PDA-AgNPs nanocomposite coatings on porous titanium. The antibacterial test, cytotoxicity test, hypodermic implantation and osteogenesis test revealed that the homogeneous PDA-AgNPs nanocomposite coating achieved the balance between the antibacterial ability and cytocompatibility, and had the best outcomes for soft tissue healing and bone formation around the implants. This study provides a facile strategy for preparing silver-loaded surfaces with both good antibacterial effect and favorable cytocompatibility, which is expected to further improve the therapeutic efficacy of silver composite-coated dental implants.

Case report: Blood purification effectively relieves multiple system failure in patient with rabies.

Rabies is an infectious disease of animal origin with a high mortality rate. In the early stages of rabies, the rabies virus (RABV) is usually undetectable in saliva and cerebrospinal fluid (CSF). In addition, there are still no effective drugs and treatments. Here, we present a case in which blood purification alleviated multisystem failures. The patient was a 45-year-old woman who presented with the fear of water and wind, restlessness, and hyperactivity. RABV was detected in her saliva by high-throughput sequencing Next Generation Sequencing (NGS) and polymerase chain reaction (PCR). Based on typical clinical symptoms and the result of NGS and PCR, the patient was diagnosed as a confirmed case of rabies. Hemodialysis combined with antiviral therapy and intensive care unit (ICU) treatment can effectively relieve circulatory failure, respiratory failure, and renal failure. Finally, she died of brain death on the 34th day of admission. The case report showed that blood purification was positive for rabies-induced organ failure. Blood purification combined with antiviral therapy can prolong the lives of patients with rabies to some extent.

Multifunctional Coatings of Titanium Implants Toward Promoting Osseointegration and Preventing Infection: Recent Developments.

Titanium and its alloys are dominant material for orthopedic/dental implants due to their stable chemical properties and good biocompatibility. However, aseptic loosening and peri-implant infection remain problems that may lead to implant removal eventually. The ideal orthopedic implant should possess both osteogenic and antibacterial properties and do proper assistance to in situ inflammatory cells for anti-microbe and tissue repair. Recent advances in surface modification have provided various strategies to procure the harmonious relationship between implant and its microenvironment. In this review, we provide an overview of the latest strategies to endow titanium implants with bio-function and anti-infection properties. We state the methods they use to preparing these efficient surfaces and offer further insight into the interaction between these devices and the local biological environment. Finally, we discuss the unmet needs and current challenges in the development of ideal materials for bone implantation.

Poly-l-lysine/Sodium Alginate Coating Loading Nanosilver for Improving the Antibacterial Effect and Inducing Mineralization of Dental Implants.

In recent years, antibacterial surface modification of titanium (Ti) implants has been widely studied in preventing implant-associated infection for dental and orthopedic applications. The purpose of this study was to prepare a composite coating on a porous titanium surface for infection prevention and inducing mineralization, which was initialized by deposition of a poly-l-lysine (PLL)/sodium alginate(SA)/PLL self-assembled coating, followed by dopamine deposition, and finally in situ reduction of silver nanoparticles (AgNPs) by dopamine. The surface zeta potential, SEM, XPS, UV-vis, and water contact angle analyses demonstrate that each coating was successfully prepared after the respective steps and that the average sizes of AgNPs were 20-30 nm. The composite coating maintained Ag+ release for more than 27 days in PBS and induced mineralization when incubated in SBF. The antibacterial results showed that the composite coating inhibited/killed bacteria on the material surface and killed bacteria around them. In addition, although this coating inhibited the initial adhesion of osteoblasts, the mineralized surface greatly enhanced the cytocompatibility. Thus, we concluded that the composite coating could prevent bacterial infections and facilitate mineralization in vivo in the early postoperative period, and then, the mineralized surface could enhance the cytocompatibility.

Immobilization of serum albumin and peptide aptamer for EPC on polydopamine coated titanium surface for enhanced in-situ self-endothelialization.

Restenosis and thrombosis are two major complications associated with vascular stents and grafts. The homing of circulating endothelial progenitor cells (EPCs) onto implant surfaces brings a new strategy to solve these problems by accelerating self -endothelialization in situ. Peptide aptamers with high affinity and specific recognition of EPCs can be immobilized to capture EPCs from the circulating blood. In this study, a biotinylated peptide aptamer (TPSLEQRTVYAK-GGGC-K-Biotin) for EPC, and bovine serum albumin (BSA) were co-immobilized onto titanium surface through avidin-biotin recognition to endow the surface with specific affinity for EPC and anti-platelet adhesion properties. Quartz crystal microbalance with dissipation (QCM-D), X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM) and water contact angle measuring were adopted for coating characterization. EPC affinity and hemocompatibility of the coating were also investigated in vitro. The results demonstrated that aptamer and BSA co-immobilized surface significantly reduced platelet adhesion and fibrinogen adsorption/activation. Besides, such functional surface could remarkably enhance EPC adhesion, without affecting the behavior of endothelial cells (ECs) and smooth muscle cells (SMCs) obviously. The result shows the possibility of utilizing such a multifunctional surface in cardiovascular implants.

[Effect of dopamine on the activity of matrix metalloproteinases and degradation of dentin collagen].

OBJECTIVE: To investigate the inhibition effect of dopamine on the activity of matrix metalloproteinases (MMP) and the effect of dopamine on degradation of dentin collagen for its potential use in caries treatment and dentin adhesive. METHODS: In the experiment of MMP activity test, 2.0 g/L dopamine + 1.0 g/L highly purified collagenase type VIII from Clostridium histolyticum served as the experimental group, and deionized water + 1.0 g/L highly purified collagenase type VIII from Clostridium histolyticum served as the negative control group, and 2% chlorhexidine + 1.0 g/L highly purified collagenase type VIII from Clostridium histolyticum served as the positive control group, and the mixture volume ratio of the two ingredients in every group was 1:9. After 15 minutes, the enzyme activity of each sample was tested by MMP activity colerimetric quantitative detection kits, and the test was repeated 5 times in each group. In the experiment of collagen degradation, the dentin slices were demineralized with 37% phosphoric acid for 1 min. In sequence, 2 dentin slices were used to observe the morphology, and the remaining 30 dentine slices were randomly divided into three groups (n = 10) according to random number table: the negative control ones were stored in 100 µl deionized water and 900 µl collagenase (7 days, 37 °C), the positive control ones were stored in 100 µl chlorhexidine and 900 µl collagenase (7 days, 37 °C) and the experimental specimens were stored in 100 µl dopamine and 900 µl collagenase (7 days, 37 °C). The degraded collagen was investigated by assaying hydroxyproline. The framework of collagen was evaluated with field emission scanning electron microscope (FE-SEM). RESULTS: The statistical results of completely random design ANOVA showed that the MMP activity and the amount of degraded collagen of the negative control group [(0.089 ± 0.011) µmol · min⁻¹ · mg⁻¹ and (2 837 ± 201) µg/cm²] were significantly higher than those of the positive control group [(0.038 ± 0.006) µmol · min⁻¹ · mg⁻¹ and (1 288 ± 172) µg/cm²] and the experimental group [(0.030 ± 0.009) µmol · min⁻¹ · mg⁻¹ and (1 389 ± 255) µg/cm²] (P < 0.05). SEM observation indicated that the structural integrity of the collagen network on dentin still existed in experiment samples and positive control groups, however, collagen fibrils were destructed and the structural integrity disappeared in the negative control groups. CONCLUSIONS: Dopamine may inhibit MMP activity and reduce the amount of degraded collagen.

A Direct Electric Field-Aided Biomimetic Mineralization System for Inducing the Remineralization of Dentin Collagen Matrix.

This in vitro study aimed to accelerate the remineralization of a completely demineralized dentine collagen block in order to regenerate the dentinal microstructure of calcified collagen fibrils by a novel electric field-aided biomimetic mineralization system in the absence of non-collagenous proteins. Completely demineralized human dentine slices were prepared using ethylene diamine tetraacetic acid (EDTA) and treated with guanidine hydrochloride to extract the bound non-collagenous proteins. The completely demineralized dentine collagen blocks were then remineralized in a calcium chloride agarose hydrogel and a sodium hydrogen phosphate and fluoride agarose hydrogel. This process was accelerated by subjecting the hydrogels to electrophoresis at 20 mA for 4 and 12 h. X-ray diffraction (XRD), scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), and transmission electron microscopy (TEM) were used to evaluate the resultant calcification of the dentin collagen matrix. SEM indicated that mineral particles were precipitated on the intertubular dentin collagen matrix; these densely packed crystals mimicked the structure of the original mineralized dentin. However, the dentinal tubules were not occluded by the mineral crystals. XRD and EDX both confirmed that the deposited crystals were fluorinated hydroxyapatite. TEM revealed the existence of intrafibrillar and interfibrillar mineralization of the collagen fibrils. A novel electric field-aided biomimetic mineralization system was successfully developed to remineralize a completely demineralized dentine collagen matrix in the absence of non-collagenous proteins. This study developed an accelerated biomimetic mineralization system which can be a potential protocol for the biomineralization of dentinal defects.

New strategies for developing cardiovascular stent surfaces with novel functions (Review).

In this review, the authors summarize the developments in surface modification of cardiovascular materials especially in author's laboratory. The authors focus on three different strategies to construct multifunctional surfaces including coimmobilization of various biomolecules on stent surfaces, stem cell based therapy systems, and a single-molecule multipurpose modification strategy in vascular interventional therapy. The roles of various molecules like heparin, gallic acid, various aptamers, and nitric oxide are highlighted in the new strategies for developing cardiovascular stent surfaces with novel functions including excellent hemocompatibility, inhibiting smooth muscle cells proliferation, and native endothelium regeneration. The success of these multifunctional surfaces provides the tremendous potential in designing the next generation of vascular stents.

Immobilization of the direct thrombin inhibitor-bivalirudin on 316L stainless steel via polydopamine and the resulting effects on hemocompatibility in vitro.

Bivalirudin (BV), a peptidic direct thrombin inhibitor, derived from hirudin, has gained increasing interest in clinical anticoagulant therapy in the recent years. In this work, a hemocompatible surface was prepared by immobilization of BV on 316L stainless steel (SS) using a bonding layer of polydopamine (DA). X-ray photoelectron spectroscopy (XPS) was used to determine the chemical composition of the surfaces to characterize polydopamine intermediate layer and the immobilized BV. The quantity of bound BV was measured by quartz crystal microbalance (QCM). The hemocompatibility in vitro was evaluated by coagulating time of activated partial thromboplastin time (aPTT) and prothrombin time (PT) assay, platelet adhesion and activation, fibrinogen adsorption, and activation and whole blood test. The effect of sterilizing method on the bioactivity of immobilized BV was also evaluated. The results showed that BVs were successfully immobilized on SS surface with the DA interlayer at a density of 98 ng/cm(2) . BV coating surface prolonged aPTT and PT, inhibited the activation of platelet and fibrinogen significantly. Sterilization by ultraviolet radiation was possible with only marginal loss of activity. Thus, the approach described here may provide a basis for the preparation of 316L SS surface modification for use in cardiovascular implants.

Biofunctionalization of titanium with PEG and anti-CD34 for hemocompatibility and stimulated endothelialization.

Thrombosis and restenosis are the main causes of failure of cardiovascular and other blood-contacting biomedical devices. It is recognized that rapid endothelialization is a promising method for preventing these complications. Convincing evidence in vivo has further emerged that the vascular homing of endothelial progenitor cells (EPCs) contributes to rapid endothelial regeneration. This study deals with improving the hemocompatibility and enhancing EPC colonization of titanium by covalently bonding PEG(600) or PEG(4000), then end-grafting of an anti-CD34 antibody. For this, a chemically hydroxylated titanium surface was aminosilanized, which was further used for covalent grafting of polyethylene glycol and the antibody. The grafting efficiency was verified in each step. In vitro platelet adhesion analysis confirmed superior hemocompatibility of the modified surface over the control. Affinity of EPC to the surface and inhibition of smooth muscle cell adhesion, two prerequisites for endothelialization, are demonstrated in in vitro cell culture. While the coating selectively stimulates EPC adhesion, its antifouling properties prevent formation of an extracellular matrix and proliferation of the cells. Additional affinity for matrix proteins in the coating is considered for further studies. Potent inhibitory effect on macrophage activation and the relative stability of the coating render this technique applicable.

Collagen/heparin coating on titanium surface improves the biocompatibility of titanium applied as a blood-contacting biomaterial.

Thrombosis and restenosis are the main causes leading to failure of cardiovascular and other blood-contacting biomedical devices. It is recognized that rapid re-endothelialization is a promising method for preventing these complications. This article deals with improving the endothelial progenitor cell (EPC) compatibility and hemocompatibility of titanium by coating an extracellular matrix-like film with heparin(hep) and collagen(col) by a layer-by-layer (LBL) self-assembly technique. In the work described here, LBL-produced col/hep coating growth is initialized by deposition of a layer of poly-L-lysine on a titanium surface, which is negatively charged after treatment with NaOH, followed by formation of a multilayer film formed by alternating deposition of negatively charged heparin and positively charged collagen using electrostatic interaction. The X-ray photoelectron spectroscopy results and fluorescence staining of collagen show that collagen is predominant on the surface and that collagen interpenetrates the heparin layer. In vitro EPC attachment and proliferation increase greatly on the col/hep coating. Immunofluorescent staining of cytoskeleton actin reveals that cells on the col/hep coating form a compact confluent cell layer after culture for 3 days. After culture for 5 days, cell viability on the col/hep increases persistently and on titanium the cell viability begins to decrease, showing that the coating possesses the ability to maintain cell viability. Platelet adhesion under dynamic conditions in vitro implies that the hemocompatibility of the col/hep coating is superior to that of titanium. The col/hep coating improves the biocompatibility of titanium and has good potential for application in blood-contacting biomaterials.

Oriented immobilization of anti-CD34 antibody on titanium surface for self-endothelialization induction.

Inducing spontaneous endothelialization of synthetic cardiovascular implant in vivo is thought to be a promising approach to solve the surface-induced thrombosis and restenosis problem. In the present study, anti-CD34 antibody, a kind of special marker of EPC, was oriented immobilized on titanium surface by means of a layer-by-layer self-assembly coating technique. The multilayer coating was prepared by first depositing one layer of avidin on the NaOH-treated titanium substrate, then depositing a layer of biotinylated protein A binding to the avidin, and finally anti-CD34 antibody was oriented immobilized by protein A binding to the Fc fragment (COOH-terminal of a antibody molecule, which has no antigen binding sites) of the anti-CD34 antibody with its antigen binding fragment (Fab) away from the titanium surface. The coated titanium was exposed to EPC derived from mouse bone marrow in vitro, and implanted into dog femoral arteries. The results suggested that the anti-CD34 antibody immobilized surfaces could increase EPC attachment and capture, and induce rapid complete endothelialization of the lumenal surface of the implant in vivo. It suggests that the approach described here may be used for fabrication of titanium-based vascular implant surfaces for inducing endothelialization in vivo.