The current status of stimuli-responsive nanotechnologies on orthopedic titanium implant surfaces.

With the continuous innovation and breakthrough of nanomedical technology, stimuli-responsive nanotechnology has been gradually applied to the surface modification of titanium implants to achieve brilliant antibacterial activity and promoted osteogenesis. Regarding to the different physiological and pathological microenvironment around implants before and after surgery, these surface nanomodifications are designed to respond to different stimuli and environmental changes in a timely, efficient, and specific way/manner. Here, we focus on the materials related to stimuli-responsive nanotechnology on titanium implant surface modification, including metals and their compounds, polymer materials and other materials. In addition, the mechanism of different response types is introduced according to different activation stimuli, including magnetic, electrical, photic, radio frequency and ultrasonic stimuli, pH and enzymatic stimuli (the internal stimuli). Meanwhile, the associated functions, potential applications and developing prospect were discussion.

Effects of vitamin C on osteoblast proliferation and osteosarcoma inhibition using plasma coated hydroxyapatite on titanium implants.

Plasma-sprayed hydroxyapatite (HAp) coated titanium (Ti) implants are being extensively used in orthopedic surgeries and post-tumor resection to repair load-bearing segmental bone defects. In this study, vitamin C, an abundantly available natural biomolecule, is loaded onto plasma-sprayed HAp-coated commercially pure titanium (cpTi) surface to evaluate its chemopreventive and osteogenic properties, suggesting its clinical significance as an alternative or adjunct therapy in the treatment for osteosarcoma bone resection. Controlled release of vitamin C from HAp coated cpTi implant is assessed by in vitro drug release study, where Korsmeyer-Peppas model was applied to understand the release kinetics. After 21 days, the implants loaded with 400 and 800 µg of vitamin C showed a cumulative release of 62.7 and 74.1% in acidic microenvironment, whereas, 50.9% and 53.1% of total vitamin C release were observed by the implants loaded with 400 and 800 µg of vitamin C in physiological pH, respectively. To observe the effects of in vitro vitamin C release on osteosarcoma and osteoblast cellular activity, MG-63 (human osteosarcoma) and hFOB (human fetal osteoblast) cells were cultured on the surface of the implant and MTT cell viability assay and FESEM were carried out at 3 and 7 days of culture. Presence of high dosages 25 mM vitamin C shows a statistically significant (p≤0.05) decrease in osteosarcoma cell viability after 3 days, while both 5 mM and 25mM vitamin C reduced cellular viability by 2.5 folds (p≤0.05) compared to the control after 7 days. Interestingly, the presence of vitamin C showed no obvious signs of cytotoxicity towards osteoblast cell-line at day 3 and day 7, as confirmed by the MTT assay. Additionally, the FESEM images depict layers of hFOB cellular morphology on the surface of the implants, suggesting excellent cytocompatibility towards the osteoblast cells. These results suggest that vitamin C loaded HAp coated cpTi implant with improved osteogenic and chemopreventive properties can be considered as a promising reconstructive option to repair the post-tumor resection defects in osteosarcoma.

Electrical Responsive Coating with a Multilayered TiO2-SnO2-RuO2 Heterostructure on Ti for Controlling Antibacterial Ability and Improving Osseointegration.

The bacterial infection and poor osseointegration of Ti implants could significantly compromise their applications in bone repair and replacement. Based on the carrier separation ability of the heterojunction and the redox reaction of pseudocapacitive metal oxides, we report an electrically responsive TiO2-SnO2-RuO2 coating with a multilayered heterostructure on a Ti implant. Owing to the band gap structure of the TiO2-SnO2-RuO2 coating, electron carriers are easily enriched at the coating surface, enabling a response to the endogenous electrical stimulation of the bone. With the formation of SnO2-RuO2 pseudocapacitance on the modified surface, the postcharging mode can significantly change the surface chemical state of the coating due to the redox reaction, enhancing the antibacterial ability and osteogenesis-related gene expression of the human bone marrow mesenchymal stem cells. Owing to the attraction for Ca2+, only the negatively postcharged SnO2@RuO2 can promote apatite deposition. The in vivo experiment reveals that the S-SnO2@RuO2-NP could effectively kill the bacteria colonized on the surface and promote osseointegration with the synostosis bonding interface. Thus, negatively charging the electrically responsive coating of TiO2-SnO2-RuO2 is a good strategy to endow modified Ti implants with excellent antibacterial ability and osseointegration.

Characterization of Micro-Threaded Stem Taper Surfaces of Cementless Hip Endoprostheses.

We investigated micro-threaded stem taper surface and its impact on premature failures, aseptic loosening, and infection in cementless hip endoprostheses. Our study focused on the fretting, and crevice corrosion of micro-threaded tapers, as well as the characterization of the microstructure and surface properties of two new and three retrieved Zweymüller stem tapers. The retrieved samples were selected and examined based on the head-stem taper interface being the sole source of modularity with a metallic component, specifically between the Ti alloy taper stem and the ceramic head. To determine the surface chemistry and microstructures of both new and retrieved hip endoprostheses stem taper titanium alloy, scanning -electron microscopy (SEM) was employed for morphological and microstructural analyses. Energy dispersive spectroscopy (EDS) was utilized for characterizing chemical element distribution, and electron backscattered diffraction (EBSD) was used for phase analysis. The roughness of the micro-threated stem tapers from different manufacturers was investigated using an optical profilometer, with standard roughness parameters Ra (average surface roughness) and Rz (mean peak to valley height of the roughness profile) being measured. Electrochemical studies revealed no fretting corrosion in retrieved stem tapers with ceramic heads. Consequently, three retrieved tapers and two new ones for comparison underwent potentiodynamic measurements in Hank's solution to determine the corrosion rate of new and retrieved stem taper surfaces. The results showed a low corrosion rate for both new and prematurely failed retrieved samples due to aseptic loosening. However, the corrosion rate was higher in infected and low-grade infected tapers. In conclusion, our study suggests that using ceramic heads reduces taper corrosion and subsequently decreases the incidence of premature failures in total hip arthroplasty.

Core-Shell NiS@SrTiO3 Nanorods on Titanium for Enhanced Osseointegration via Programmed Regulation of Bacterial Infection, Angiogenesis, and Osteogenesis.

Developing bone implants with dynamic self-adjustment of antibacterial, angiogenic, and osteogenic functions in line with a bone regenerative cascade is highly required in orthopedics. Herein, a unique core-shell nanorods array featuring a thin layer of NiS coated on each SrTiO3 nanorod (NiS@SrTiO3) was in situ constructed on titanium (Ti) through a two-step hydrothermal treatment. Under near-infrared (NIR) irradiation, the photoresponsive effect of NiS layer in synergy with the physical perforation of SrTiO3 nanorods initially enabled in vitro antibacterial rates of 96.5% to Escherichia coli and 93.1% to Staphylococcus aureus. With the degradation of the NiS layer, trace amounts of Ni ions were released, which accelerated angiogenesis by upregulating the expression of vascular regeneration-related factors, while the gradual exposure of SrTiO3 nanorods could simultaneously enhance the surface hydrophilicity in favor of cell adhesion and slowly release Sr ions to promote the proliferation and differentiation of MC3T3-E1 cells. The in vivo assessment verified not only the satisfactory antibacterial effect but also the superior osteogenic ability of the NiS@SrTiO3/Ti group with the aid of NIR irradiation, finally promoting the osseointegration of the Ti implant. The modification method endowing Ti implant with antibacterial, angiogenic, and osteogenic functions provides a new strategy to improve the long-term reliability of Ti-based devices.

Simultaneous acceleration of osteogenesis and angiogenesis by surface oxygen vacancies of rutile nanorods.

Advanced implants with simultaneous accelerated osteogenic and angiogenic capacities are of great importance for osteointegration. Much attention has been paid to simultaneously enhancing the osteogenesis and angiogenesis by surface decoration of bioactive molecules or ions on biomaterial surface, but the inherent physical cue of material surface down to the atomic-scale features have always been ignored. In this study, we demonstrate that regulation of surface oxygen vacancies defects of rutile nanorods are able to simultaneous accelerate the osteogenesis and angiogenesis. The concentration of surface oxygen vacancies defects of rutile nanorods can be manipulated by simple redox processing. The osteogenic differentiation of mesenchymal stem cells (MSCs), angiogenic differentiation and vessel-like tube structures of human umbilical vein endothelial cells (HUVECs) on oxygen vacancies rich surface are significantly up-regulated. This work therefore emphasizes the critical role of the inherent material atomic-scale features and provides a novel strategy to accelerate the osteogenesis and angiogenesis of Ti-based implant.

TiO2 Nanotopography-Driven Osteoblast Adhesion through Coulomb's Force Evolution.

Nanotopography is an effective method to regulate cells' behaviors to improve Ti orthopaedic implants' in vivo performance. However, the mechanism underlying cellular matrix-nanotopography interactions that allows the modulation of cell adhesion has remained elusive. In this study, we have developed novel nanotopographic features on Ti substrates and studied human osteoblast (HOb) adhesion on nanotopographies to reveal the interactive mechanism regulating cell adhesion and spreading. Through nanoflat, nanoconvex, and nanoconcave TiO2 nanotopographies, the evolution of Coulomb's force between the extracellular matrix and nanotopographies has been estimated and comparatively analyzed, along with the assessment of cellular responses of HOb. We show that HObs exhibited greater adhesion and spreading on nanoconvex surfaces where they formed super matured focal adhesions and an ordered actin cytoskeleton. It also demonstrated that Coulomb's force on nanoconvex features exhibits a more intense and concentrated evolution than that of nanoconcave features, which may result in a high dense distribution of fibronectin. Thus, this work is meaningful for novel Ti-based orthopaedic implants' surface designs for enhancing their in vivo performance.

Bioinspired TiO2/chitosan/HA coatings on Ti surfaces: biomedical improvement by intermediate hierarchical films.

The most common reasons for hard-tissue implant failure are structural loosening and prosthetic infections. Hence, in this study, to overcome the first problem, different bioinspired coatings, including dual acid-etched, anodic TiO2nanotubes array, anodic hierarchical titanium oxide (HO), micro- and nanostructured hydroxyapatite (HA) layers, and HA/chitosan (HA/CS) nanocomposite, were applied to the titanium alloy surfaces. X-ray diffraction and FTIR analysis demonstrated that thein situHA/CS nanocomposite formed successfully. The MTT assay showed that all samples had excellent cell viability, with cell proliferation rates ranging from 120% to 150% after 10 days. The HO coating demonstrated superhydrophilicity (θ≈ 0°) and increased the wettability of the metallic Ti surface by more than 120%. The friction coefficient of all fabricated surfaces was within the range of natural bone's mechanical behavior. The intermediate HO layer increased the adhesion strength of the HA/CS coating by more than 60%. The HO layer caused the mechanical stability of HA/CS during the 1000 m of friction test. The microhardness of HA/CS (22.5 HV) and micro-HA (25.5 HV) coatings was comparable to that of human bone. A mechanism for improved adhesion strength of HA/CS coatings by intermediate oxide layer was proposed.

A facile chemical process to form an ultrathin hydroxyapatite layer with a customizable silver-releasing function on a titanium implant.

Silver-containing hydroxyapatite (Ag/HAp) layer on a bioinert material provides both bioactive and antibacterial properties; however, the Ag release duration needs to be customized to a patient's age and metabolism for minimizing the toxic effects. Herein, we present a facile chemical process to produce an ultrathin Ag/HAp layer on a Ti implant with a customized Ag-releasing profile. The process involves the following steps: preparation of a slurry-type reagent by mixing calcium phosphate powder with an aqueous AgNO3 solution, burying a Ti substrate in the slurry, and heating the slurry in air. An HAp layer, approximately 50 nm thick, with Ag particle deposits was obtained using this process. The Ag-particle content can be varied by adjusting the concentration of AgNO3 solution used for slurry preparation, resulting in different Ag-release profiles in a physiological solution. For instance, Ag release was retained for up to 30 days when 100 mM AgNO3 was used, whereas the release lasted 15 days when 10 mM AgNO3 was used. The duration of the antibacterial activity varied accordingly, but Ag-release-derived cytotoxicity was not observed irrespective of the AgNO3 concentration. In addition, differentiation of osteoblast-like cells was facilitated owing to the formation of the HAp layer. Thus, the chemical process presented in this study allows the production-at a clinical site-of an Ag/HAp layer customized to the patient's needs.

The response of macrophages and their osteogenic potential modulated by micro/nano-structured Ti surfaces.

Current understanding on the interactions between micro/nano-structured Ti surfaces and macrophages is still limited. In this work, TiO2 nano-structures were introduced onto acid-etched Ti surfaces by alkali-heat treatment, ion exchange and subsequent heat treatment. By adjusting the concentration of NaOH during alkali-heat treatment, nano-flakes, nano-flakes mixed with nano-wires or nano-wires could formed on acid-etched Ti surfaces. The micro- and micro/nano-structured Ti surfaces possessed similar surface chemical and phase compositions. In vitro results indicate that the morphology of macrophages was highly dependent on the morphological features of nano-structures. Nano-flakes and nano-wires were favorable to induce the formation of lamellipodia and filopodia, respectively. Compared to micro-structured Ti surface, micro/nano-structured Ti surfaces polarized macrophages to their M2 phenotype and enhanced the gene expressions of osteogenic growth factors in macrophages. The M2 polarized macrophages promoted the maturation of osteoblasts. Compared to that with nano-flakes or nano-wires, the surface with mixed features of nano-flakes and nano-wires exhibited stronger anti-inflammatory and osteo-immunomodulatory effects. The findings presented in the current work suggest that introducing micro/nano-topographies onto Ti-based implant surfaces is a promising strategy to modulate the inflammatory response and mediate osteogenesis.

Application of Natural Neutrophil Products for Stimulation of Monocyte-Derived Macrophages Obtained before and after Osteochondral or Bone Injury.

We evaluated the use of some neutrophil products, namely; autologous rabbit antimicrobial neutrophil extract (rANE), heterologous porcine antimicrobial neutrophil extract (pANE), neutrophil degranulation products (DGP) and neutrophil microvesicles (MVs) for stimulation of monocyte-derived macrophages (MDMs) to improve healing. Two animal models were evaluated; the rabbit model for autologous osteochondral transplantation (OT) with application of rabbit ANE, DGP or MVs for MDMs stimulation, and the ovine model of the insertion of a Ti implant with the use of porcine ANE, and ovine DGP or MVs for MDMs stimulation. Macrophage activity was assessed on the basis of free radical generation and arginase activity. We estimated that DGP acted in a pro-inflammatory way both on rabbit and ovine MDMs. On the other hand, MVs acted as anti-inflammatory stimulator on MDMs in both experiments. The response to ANE depended on origin of extract (autologous or heterologous). Macrophages from rabbits before and after OT stimulated with autologous extract generated lower amount of NO and superoxide, especially after transplantation. In the ovine model of Ti implant insertion, heterologous ANE evoked increased macrophage pro-inflammatory activity. Our study revealed that these neutrophil products could regulate activity of macrophages, polarizing them into pro-or anti-inflammatory phenotypes that could enhance bone and osteochondral tissue healing.

Decontamination of Titanium Surface Using Different Methods: An In Vitro Study.

Contamination of implants is inevitable during different steps of production as well as during the clinical use. We devised a new implant cleaning strategy to restore the bioactivities on dental implant surfaces. We evaluated the efficiency of the Finevo cleaning system, and Ultraviolet and Plasma treatments to decontaminate hydrocarbon-contaminated titanium disks. The surfaces of the contaminated titanium disks cleaned using the Finevo cleaning system were similar to those of the uncontaminated titanium disks in scanning electron microscopy and X-ray photoelectron spectroscopy analysis, but no obvious change in the roughness was observed in the scanning probe microscopy analysis. The rat bone marrow mesenchymal stem cells (rBMMSCs) cultured on the treated titanium disks attached to and covered the surfaces of disks cleaned with the Finevo cleaning system. The alkaline phosphatase activity, calcium deposition, and osteogenesis-related gene expression in rBMMSCs on disks cleaned using the Finevo cleaning system were higher compared to those in the ultraviolet and plasma treatments, displaying better cell functionality. Thus, the Finevo cleaning system can enhance the attachment, differentiation, and mineralization of rBMMSCs on treated titanium disk surfaces. This research provides a new strategy for cleaning the surface of contaminated titanium dental implants and for restoration of their biological functions.

Micro/nanostructured TiO2/ZnO coating enhances osteogenic activity of SaOS-2 cells.

Although titanium implants account for a large proportion of the commercial dental market, their bioactivity are inadequate in many applications. A micro- and nano- scale hierarchical surface topography of the implant is suggested for rapid osseointegration from the biomimetic perspective. Moreover, Zinc (Zn) is an essential element in the skeletal system. Thus, a micro/nanostructured TiO2/ZnO coating, produced by micro-arc oxidation, and hydrothermal treatment, and heat treatment, was designed to endow the implant surface with enhanced osteogenic capacity. Physiochemical properties and biological effects of this coating were investigated in our study. The annealed micro/nanostructured TiO2/ZnO coating exhibited higher hydrophilicity and fibronectin adsorption ability compared to the micro-arc oxidation modified TiO2 coating. SaOS-2 cells grown on the annealed micro/nanostructured TiO2/ZnO coating showed increased alkaline phosphatase activity and collagen secretion, and immunofluorescence labeling revealed an upregulation of osteopontin, collagen type ι and osteocalcin. The micro/nanostructure and incorporation of Zn were considered to perform positive effect on the enhanced osteogenic activity of SaOS-2 cells. In conclusion, the micro/nanostructured TiO2/ZnO structure is simple, stable, and easy to produce and scale up, has promising applications in the surface modification of titanium implants.

Verification of osteoblast differentiation on airborne-particle abrasion, large-grit, acid-etched surface of titanium implants regulated by yes-associated protein and transcriptional coactivator with PDZ-binding motif.

Although airborne-particle abrasion, large-grit, acid-etched (SLA) surface technology can promote implant osseointegration; its mechanism remains unclear. By preparing the SLA titanium (Ti) plate (SLA Ti) and Polished Ti plate (Polished Ti), this experiment investigates the expression and distribution of the Yes-associated protein (YAP) and transcriptional coactivator with the PDZ-binding motif (TAZ) in MC3T3-E1 cells. In addition, gene YAP and TAZ silencing on the SLA Ti was conducted to observe changes in the osteoblast differentiation markers, runt-related transcription factor-2 (Runx2) and bone sialoprotein (BSP). The results demonstrated that SLA Ti surface microtopography could induce YAP/TAZ's transfer from the cytoplasm to the nuclei of MC3T3-E1 cells. The expression of YAP/TAZ increased in terms of mRNA and protein. After silencing the YAP/TAZ genes, Runx2 and BSP decreased, suggesting that YAP/TAZ plays an important regulatory role in this process. Meanwhile, the results also showed that SLA microtopography enhanced the expression of integrins α1, α2, and ß1. After silencing the integrin α1, α2, and ß1 genes, YAP and TAZ decreased in terms of mRNA and protein. Therefore, this experiment was the first to confirm that SLA surface microtopography facilitates osteoblast differentiation by regulating YAP/TAZ and confirms that the process can be related to integrins α1, α2, and ß1.

Bone mesenchymal stem cell secretion of sRANKL/OPG/M-CSF in response to macrophage-mediated inflammatory response influences osteogenesis on nanostructured Ti surfaces.

Although it has been well established that osteogenic differentiation of bone mesenchymal stem cells (bMSCs) as well as osteoclastic differentiation of macrophages can be manipulated by the nanostructure of biomaterial surfaces, the interactions among the effects of the surface on immune cells and bMSCs remained unknown. Therefore, in this study, the osteogenic behaviors and secretion of osteoclastogenesis-related cytokines of human bMSCs on TiO2 nanotubular (NT) surfaces in conditioned medium (CM) generated by macrophages cultured on the respective NT surfaces (NT-CM) were analyzed. Although bMSCs showed consistent osteogenic behaviors on the NT5 and NT20 surfaces in both standard culture medium and both types of NT-CM, collagen synthesis and extracellular matrix mineralization were partially impeded on the NT20 surface in NT20-CM and bMSC cytokine secretions on the NT20 surface in NT20-CM elicited remarkable multinuclear giant cell and osteoclast formation compared with that observed on the NT5 surface in NT5-CM. After implantation in vivo, mineralized bone formation was significantly delayed around the NT20 implant compared with the NT5 implant, but both surfaces contributed to good bone formation after 12 weeks. The results obtained in this study advance our understanding of the confounding influence of the implant surface nanostructure, macrophage inflammatory response, and osteogenic differentiation of bMSCs as well as the retro-regulative effects of bMSCs on the osteoclastic differentiation of macrophages, and the culture system based on different NT surfaces and CM generated on the respective surfaces may provide a systematic research model for evaluating the performance of endosseous implants as well as a prospective approach for improving implant osseointegration via immune-regulation.

Enhanced Osseointegration of Hierarchically Structured Ti Implant with Electrically Bioactive SnO2-TiO2 Bilayered Surface.

The poor osseointegration of Ti implant significantly compromise its application in load-bearing bone repair and replacement. Electrically bioactive coating inspirited from heterojunction on Ti implant can benefit osseointegration but cannot avoid the stress shielding effect between bone and implant. To resolve this conflict, hierarchically structured Ti implant with electrically bioactive SnO2-TiO2 bilayered surface has been developed to enhance osseointegration. Benefiting from the electric cue offered by the built-in electrical field of SnO2-TiO2 heterojunction and the topographic cue provided by the hierarchical surface structure to bone regeneration, the osteoblastic function of basic multicellular units around the implant is significantly improved. Because the individual TiO2 or SnO2 coating with uniform surface exhibits no electrical bioactivity, the effects of electric and topographic cues to osseointegration have been decoupled via the analysis of in vivo performance for the placed Ti implant with different surfaces. The developed Ti implant shows significantly improved osseointegration with excellent bone-implant contact, improved mineralization of extracellular matrix, and increased push-out force. These results suggest that the synergistic strategy of combing electrical bioactivity with hierarchical surface structure provides a new platform for developing advanced endosseous implants.

Ion-Doped Silicate Bioceramic Coating of Ti-Based Implant.

Titanium and its alloy are known as important load-bearing biomaterials. The major drawbacks of these metals are fibrous formation and low corrosion rate after implantation. The surface modification of biomedical implants through various methods such as plasma spray improves their osseointegration and clinical lifetime. Different materials have been already used as coatings on biomedical implant, including calcium phosphates and bioglass. However, these materials have been reported to have limited clinical success. The excellent bioactivity of calcium silicate (Ca-Si) has been also regarded as coating material. However, their high degradation rate and low mechanical strength limit their further coating application. Trace element modification of (Ca-Si) bioceramics is a promising method, which improves their mechanical strength and chemical stability. In this review, the potential of trace element-modified silicate coatings on better bone formation of titanium implant is investigated.