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Sclerostin is a negative regulator of the Wnt signaling pathway for osteoblast differentiation. In this study, osteoblasts were co-cultured with osteocytes (MLO-Y 4 cells) on the surface of sclerostin antibody-conjugated TiO 2 nanotube arrays (TNTs-scl). Field emission scanning electron microscopy (SEM), contact angle measurement and confocal laser scanning microscope (CLSM) were employed to characterize the conjugation of sclerostin antibody onto the surface of TiO 2 nanotube arrays. The cellular viability and morphology results displayed TNTs-scl (TNT30-scl and TNT70-scl) were beneficial to the growth of MLO-Y4 cells. There was no apparent change in sclerostin gene expression between MLO-Y4 cells grown on TNTs and TNTs-scl. However, TNTs-scl significantly reduced the amount of sclerostin in the medium. In comparison with the control groups, osteoblasts displayed higher differentiation capability when co-cultured with MLO-Y4 cells on the surface TNTs-scl, which was indicated by the ALP activity, mineralization capability as well as expression levels of key proteins in Wnt signaling. This study provides a simple strategy to engineer titanium surface for bone fracture recovery, especially in osteoporotic conditions., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Titanium substrates with micro/nano hierarchical features could positively mediate the osteogenesis of a titanium implant; nevertheless, the underlying molecular mechanism needs to be further revealed. In this work, we fabricated a micro/nano hierarchically structured Ti (MNT) sample and attempted to evaluate its topography-mediated biological effects and potential molecular mechanisms in vitro. The results proved that MNT could not only affect cell morphology and osteogenic differentiation, but also regulate ROCK activity cell biological functions of osteoblasts involved in ROCK activation, β-catenin accumulation, and high-Wnt5a expression in respect to topographical features. Moreover, blockade of ROCK activation resulted in significant inhibition of cell differentiation and Wnt5a expression. Furthermore, the anti-Wnt5a significantly down-regulated ROCK activity. In short, these results indicate the important role of ROCK-Wnt5a feedback loop in regulating cell differentiation by topographies., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Oxidative stress is commonly existed in bone degenerative disease (osteoarthritis, osteoporosis etc.) and some antioxidants had great potential to enhance osteogenesis. In this study, we aim to investigate the anti-oxidative properties of various TiO 2 nanotubes (TNTs) so to screen the desirable size for improved osteogenesis and reveal the underlying molecular mechanism in vitro. Comparing cellular behaviors under normal and oxidative stress conditions, an interesting conclusion was obtained. In normal microenvironment, small TNTs were beneficial for adhesion and proliferation of osteoblasts, but large TNTs greatly increased osteogenic differentiation. However, after H 2 O 2 (300 μM) treatment (mimicking oxidative stress), only large TNTs samples demonstrated superior cellular behaviors of increased osteoblasts' adhesion, survival and differentiation when comparing with those of native titanium (control). Molecular results revealed that oxidative stress resistance of large nanotubes was closely related to the high expression of integrin α5β1 (ITG α5β1), which further up-regulated the production of anti-apoptotic proteins (p-FAK, p-Akt, p-FoxO3a and Bcl2) and down-regulated the expression of pro-apoptotic protein (Bax). Moreover, we found that Wnt signals (Wnt3a, Wnt5a, Lrp5, Lrp6 and β-catenin) played an important role in promoting osteogenic differentiation of osteoblasts under oxidative condition., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

To improve the biological performance of titanium substrates, a bioactive multilayered structure of chitosan/gelatin pair, containing zinc ions, was constructed via a layer-by-layer self-assembly technique. The successful preparation of zinc ions incorporated multilayer films was demonstrated by scanning electron microscopy, X-ray photoelectron spectroscopy, and contact angle measurements, respectively. The biological behaviors of osteoblasts adhered to modified Ti substrates were investigated in vitro via cytoskeleton observation, cell viability measurement, and alkaline phosphatase activity assay. The cytocompatibility evaluation verified that the present system was capable of promoting the growth of osteoblasts. In addition, Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacteria were used to evaluate antibacterial property of modified Ti substrates. Bacterial adhesion and viability assay confirmed that Zn-loaded multilayer films were able to inhibit the adhesion and growth of bacteria. The approach presented here affords an alternative to reduce bacterial infection and promote osteoblast growth for titanium-based implants., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Oxidative stress induced by reactive oxygen species (ROS) overproduction would hinder bone healing process at the interface of bone/implant, yet underlying mechanism remains to be explored. To endow titanium (Ti) substrates with antioxidant activity for enhanced bone formation, multilayered structure composing of chitosan-catechol (Chi-C), gelatin (Gel) and hydroxyapatite (HA) nanofibers was constructed on Ti substrates. Surface wettability and topography of multilayer coated Ti substrates were characterized by water contact angle measurement, scanning electron microscopy and atomic force microscopy, respectively. Chi-C containing multilayer on Ti surface effectively protected osteoblasts from ROS damage, which was revealed by high level of intracellular ROS scavenging activity and reduced oxidative damage on cellular level by regulating the expression of cell adhesion related genes (integrin αv, β3, CDH11 and CDH2). Moreover, it regulated the production of cell adhesive and anti-apoptotic related proteins (p-MYPT1, p-FAK, p-Akt and Bcl-2) and pro-apoptotic critical executioners (Bax and cleaved caspase 3). Beside, the composite multilayer of Chi-C/Gel/HA nanofibers on Ti substrates promoted osteoblasts differentiation, which was evidenced by high expression levels of alkaline phosphatase activity, collagen secretion, ECM mineralization and osteogenesis-related genes expression in vitro. The in vivo experiments of μ-CT analysis, push out test and histochemistry staining further confirmed that Chi-C multilayered implant had great potential for improved early bone healing. Overall, the study offers an effective strategy for the exploration of high quality Ti implants for orthopedic applications., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Surface microroughness plays an important role in determining osteoblast behavior on titanium. Previous studies have shown that osteoblast differentiation on microtextured titanium substrates is dependent on alpha-2 beta-1 (α2β1) integrin signaling. This study used focused ion beam milling and scanning electron microscopy, combined with three-dimensional image reconstruction, to investigate early interactions of individual cells with their substrate and the role of integrin α2β1 in determining cell shape. MG63 osteoblast-like cells on sand blasted/acid etched (SLA) Ti surfaces after 3 days of culturing indicated decreased cell number, increased cell differentiation, and increased expression of mRNA levels for α1, α2, αV, and β1 integrin subunits compared to cells on smooth Ti (PT) surfaces. α2 or β1 silenced cells exhibited increased cell number and decreased differentiation on SLA compared to wild-type cells. Wild-type cells on SLA possessed an elongated morphology with reduced cell area, increased cell thickness, and more apparent contact points. Cells on PT exhibited greater spreading and were relatively flat. Silenced cells possessed a morphology and phenotype similar to wild-type cells grown on PT. These observations indicate that surface microroughness affects cell response via α2β1 integrin signaling, resulting in a cell shape that promotes osteoblastic differentiation., (© 2014 Wiley Periodicals, Inc.)

To treat the osteoporosis and regulate the biological behaviors of both osteoblasts and osteoclasts, we prepared a natural polysaccharide-derived nanocomposite, containing alendronate-grafted hyaluronate (HA-Aln) and bone morphogenetic protein 2 (BMP-2) and investigated its synergistic regulation on the behaviors of osteoblasts and osteoclasts in vitro. The HA-Aln/BMP-2 nanocomposite was fabricated through the electrostatic interactions between the HA-Aln molecule and BMP-2 molecule. Here, BMP-2 was used to improve the osteoblast-mediated bone formation. Alendronate (Aln), a targeting ligand to bone matrix, was used to inhibit the osteoclast-mediated bone resorption. In vitro results showed that HA-Aln/BMP-2 nanocomposite could effectively maintain the bioactivity of loaded drugs. The osteoblasts that treated with the HA-Aln/BMP-2 nanocomposite presented a higher level of cell motility, alkaline phosphatase (ALP) activity, mineralization capacity, and osteoblast-related gene expressions (runt-related transcription factor 2, osterix, ALP, collagen type I, osteocalcin, and osteopontin), as compared to that of control group. Besides, the RAW264.7 cells that were treated with HA-Aln/BMP-2 nanocomposite showed a lower level of osteoclastic differentiation. Overall, the HA-Aln/BMP-2 nanocomposite exhibits promising potential as an efficient carrier for co-delivery of anti-osteoporotic drug and growth factors to promote osteoblastic differentiation and bone formation while suppressing osteoclastic activity.

Osseointegration is essential for the long-term survival of orthopedic implants. Inspired by the hierarchical structure of natural bone, we fabricated a hierarchical structure with osteoinduction potential on titanium alloy (Ti6Al7Nb) substrates via a spin-assisted layer-by-layer assembly technique, with hydroxyapatite nanofibers as the intercalated materials and gelatin and chitosan as the polycation and polyanion, respectively. The as-synthesized hydroxyapatite nanofibers were characterized using scanning electron microscopy (SEM), transmission electron microscopy, Fourier transform infrared spectroscopy and X-ray diffraction. The change of water contact angle corresponding to different layers indicated the formation of a multilayered structure, since different components have their inherent wettability natures. The multilayered lamellar structure was revealed by the cross-sectional view of SEM, suggesting that the film was successfully deposited onto Ti6Al7Nb substrates. Osteoblasts cultured on the hierarchical structure deposited Ti alloy substrates displayed significantly higher cell viability (P<0.01) and better adhesion, a higher production level of alkaline phosphatase, mineralization, genes expressions of osteocalcin and osteopontin (P<0.01 or P<0.05) compared to those of native Ti6Al7Nb substrates after culture for 4, 7 or 14days. These results indicated that the lamellar structure was beneficial for the biological functions of osteoblasts, establishing the basis for osseointegration of a titanium alloy implant., (Copyright © 2014 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

In this study, we report the influence of titania nanotubes (TiNTs) dimensions on the adsorption of collagen (COL) and fibronectin (FN), and its subsequent effect on the growth of osteoblasts. TiNTs with different diameters of around 30 and 100 nm were prepared with anodization. The adsorption profiles of proteins and cell behaviors were evaluated using spectrophotometric measurement, immunofluorescence staining, cell viability, and cytoskeleton morphology, respectively. The results showed that although the growth of osteoblasts was highly sensitive to the dimensions TiNTs, the preadsorbed COL and FN could reduce the difference. Molecular dynamics (MD) simulation results confirmed that the main driving force for protein adsorption was the physical adsorption. The TiNTs with bigger dimensions had higher interaction energies, and thus leading to higher proteins (COL and FN) adsorption and obvious influences on cell behaviors. MD simulation revealed that the orientation and conformation of proteins adsorbed onto surfaces of TiNTs was critical for cell integrins to recognize specific sites. When FN molecules adsorbed onto the surfaces of TiNTs, their RGD (Arg-Gly-Asp) sites were easily exposed to outside and more likely to bond with the fibronectin receptors, in turn regulating the cellular behaviors., (© 2013 Wiley Periodicals, Inc.)

To improve the biological performance of titanium implant, a series of Zn-incorporated coatings were fabricated on the microrough titanium (Micro-Ti) via sol-gel method by spin-coating technique. The successful fabrication of the coating was verified by combined techniques of scanning electron microscopy, surface profiler, X-ray diffraction, X-ray photoelectron spectroscopy, and water contact angle measurements. The incorporated zinc existed as ZnO, which released Zn ions in a sustained manner. The Zn-incorporated samples (Ti-Zn0.08, Ti-Zn0.16, and Ti-Zn0.24) efficiently inhibited the adhesion of both Gram-positive (Staphylococcus aureus) and Gram-negative (Pseudomonas aeruginosa) bacteria. The in vitro evaluations including cell activity, alkaline phosphatase (ALP), mineralization, osteogenic genes expressions (Runx2, ALP, OPG, Col I, OPN, and OC), and tartrate-resistant acid phosphatase, confirmed that Ti-Zn0.16 sample was the optimal one to regulate the proliferation or differentiation for both osteoblasts and osteoclasts. More importantly, in vivo evaluations including Micro-CT analysis, push-out test, and histological observations verified that Ti-Zn0.16 implants could efficiently promote new bone formation after implantation for 4 and 12 weeks, respectively. The resulting material thus has potential application in orthopedic field.

Orthopedic implants failures are generally related to poor osseointegration and/or bacterial infection in clinical application. Surface functionalization of an implant is one promising alternative for enhancing osseointegration and/or reducing bacterial infection, thus ensuring the long term survival of the implant. In this study, titanium (Ti) substrates were surface functionalized with a polydopamine (PDOP) film as an intermediate layer for post-immobilization of chitosan-lauric acid (Chi-LA) conjugate. Chi-LA conjugate was synthesized and characterized by Fourier transform infrared spectroscopy (FTIR) and hydrogen proton nuclear magnetic resonance (NMR) spectrometer, respectively. Lauric acid (LA), a natural saturated fatty acid, was used mainly due to its good antibacterial property. Scanning electron microscopy (SEM) and water contact angle measurements were employed to detect the morphology changes and surface wettability of Ti substrates. The results suggested that Chi-LA conjugate was successfully immobilized onto the surfaces of Ti substrates. In vitro tests confirmed that the cell adhesion, cell viability, intracellular alkaline phosphatase activity and mineralization capacity of osteoblasts were remarkably improved when cultured onto Chi-LA surface functionalized Ti substrates. Antibacterial assay against Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa (P. aeruginosa) showed that the Chi-LA modified Ti substrates efficiently inhibited the adhesion and growth of bacteria. Overall, this study developed a promising approach to fabricate functional Ti-based orthopedic implants, which could enhance the biological functions of osteoblasts and concurrently reduce bacteria adhesion., (Copyright © 2014 Elsevier B.V. All rights reserved.)

To investigate the influence of magnetized hydroxyapatite on the growth and differentiation of osteoblasts, hydroxyapatite (HA) and magnetized hydroxyapatite (mHA) were synthesized and characterized. The cell viability, differentiation, and morphologies of osteoblasts were investigated in vitro, respectively. The results showed that compared to HA, cells cultured with mHA had better cell viability, and both HA and mHA were beneficial to the early differentiation of osteoblasts. Furthermore, the interaction mechanism between mHA and osteoblasts was elucidated using a molecular dynamics simulation. The simulation results indicated that when cultured with osteoblasts, HA adsorbed bovine serum protein onto its surface from the medium immediately, which was beneficial to the adhesion and proliferation of osteoblasts. The main driving force for the adsorption of bovine serum was the electronic properties of HA crystal faces. The (211) crystal face of HA had the highest electron density among its all crystal faces, thus mainly contributing to the protein adsorption of HA. Nevertheless, the (211) crystal face of mHA still had a relatively higher electron density than that of HA, thus possessing better protein adsorption than that of HA, and in turn promoting the biological functions of osteoblasts., (Copyright © 2013 Elsevier B.V. All rights reserved.)

To mimic the extracellular matrix of natural bone, apatite/gelatin composite was deposited onto nanostructured titanium substrates via a coprecipitation method, which was pretreated by potassium hydroxide and heat treatment to generate an anticorrosive nanostructured layer. The successful formation of the apatite/gelatin nanocomposite onto titanium surfaces was revealed by Fourier transform infrared spectroscopy, field-emission scanning electron microscopy, atomic force microscopy (AFM), and thin film X-ray diffraction (TF-XRD) measurements, respectively. The immunofluorescence staining of vinculin revealed that the apatite/gelatin nanocomposite deposited titanium substrate was favorable for cell adhesion. More importantly, bone marrow stromal cells cultured onto the apatite/gelatin nanocomposite deposited titanium substrates displayed significantly higher (p < 0.05 or p < 0.01) proliferation and differentiation levels of alkaline phosphatase, mRNA expressions of osteocalcin (OC), osteopontin (OPN), and collagen type I (Col I), and OC content after culture for 7, 14, and 21 days, respectively, which was also revealed by the immunofluorescence analysis of OC and OPN expression. The deposition of apatite/gelatin nanocomposite improved bone density (p < 0.05) and bone-implant contact rate (p < 0.05), which was reflected by microcomputed tomography analysis and histological evaluation in vivo using a rabbit model. This work provides an approach to fabricate high-performance titanium-based implants with enhanced bone osseointegration., (Copyright © 2012 Wiley Periodicals, Inc.)

To improve the corrosion resistance and biological performance of commercially pure titanium (cp-Ti) substrates, potassium hydroxide was employed to modify the surfaces of titanium substrates, followed by biomimetic deposition of apatite on the substrates in a simulated body fluid. The morphologies of native and treated titanium substrates were characterized by field emission scanning electron microscopy (FE-SEM). Treatment with potassium hydroxide led to the formation of intermediate layers of potassium titanate on the surfaces of titanium substrates, while apatite was subsequently deposited onto the intermediate layer. The formation of potassium titanate and apatite was confirmed by thin-film X-ray diffraction and FE-SEM equipped with energy dispersive spectroscopy, respectively. Electrochemical impedance spectroscopy showed that the formed potassium titanate layer improved the corrosion-resistance properties of titanium substrates. The influence of modified titanium substrates on the biological behavior of mesenchymal stem cells (MSCs), including osteogenic differentiation, was investigated in vitro. Compared with cp-Ti substrates, MSCs cultured onto alkali- and heat-treated titanium substrates and apatite-deposited titanium substrates displayed significantly higher (P<0.05 or P<0.01) proliferation and differentiation levels of alkaline phosphatase and osteocalcin in 7 and 14day cultures, respectively. More importantly, our results suggest that the modified titanium substrates have great potential for inducing MSCs to differentiate into osteoblasts. The approach presented here may be exploited to fabricate titanium-based implants., (Copyright 2009 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Chitosan (Chi) and poly (styrene sulfonate) (PSS) were employed to surface modify titanium thin film via electrostatic self-assembly (ESA) technique in order to improve its biocompatibility. The surface chemistry, wettability and surface topography of the coated films with different number of deposited layers were investigated by using X-ray photoelectron spectroscopy (XPS), water contact angle measurement and atomic force microscopy (AFM), respectively. The results indicated that a full surface coverage for the outmost layer was achieved at least after deposition of five layers, i.e., PEI/(PSS/Chi)2 on the titanium films. The formed multi-layered structure of PEI(PSS/Chi)x (x > or = 2) on the titanium film was stable in air at room temperature and in phosphate buffered solution (PBS) for at least 3 weeks. Cell proliferation, cell viability, DNA synthesis as well as differentiation function (alkaline phosphatase) of osteoblasts on chitosan-modified titanium film (PEI/(PSS/Chi)6) and control sample were investigated, respectively. Osteoblasts cultured on chitosan-modified titanium film displayed a higher proliferation tendency than that of control (p < 0.01). Cell viability, alkaline phosphatase as well as DNA synthesis measurements indicated that osteoblasts on chitosan-modified titanium films were greater (p < 0.01) than those for the control, respectively. These results suggest that surface modification of titanium film was successfully achieved via deposition of PEI/(PSS/Chi)x layers, which is useful to enhance the biocompatibility of the titanium film.

The objective of the present study was to surface modify the titanium thin films to improve its biocompatibility. A layer-by-layer (LBL) self-assembly technique, based on the electrostatic interactions mediated adsorption of chitosan (Chi) and silk fibroin (SF), was used leading to the formation of multilayers on the titanium thin film surfaces. The surface chemistry and wettability of LBL films were investigated by X-ray photoelectron spectroscopy (XPS), water contact angle measurement, and atomic force microscopy, respectively. XPS and contact angle measurement results indicated that a full SF/Chi pair film was formed after the deposition layers of PEI/(SF/Chi)(2) on the titanium film surfaces. The topographies of multilayered films were directly related to the corresponding outmost layer components. The build-up of such SF/Chi pair films on titanium films may in turn affect the biocompatibility of the modified titanium films. Therefore, an in vitro investigation was performed to confirm this hypothesis. Cell proliferation, cell viability, DNA synthesis as well as differentiation function (alkaline phosphatase) of osteoblasts on LBL-modified titanium films and control samples were investigated, respectively. Osteoblasts cultured on modified titanium films was found to be higher proliferation tendency than that on control (p < 0.05). Cell viability, alkaline phosphatase as well as DNA synthesis measurement indicated that osteoblasts on LBL-modified films were greater (p < 0.05 or p < 0.01) than the control, respectively. These results suggest that surface engineering of titanium was successfully achieved via LBL deposition of Chi/SF pairs, and enhanced its cell biocompatibility. The approach presented in the study may be exploited as an efficient alternative for surface engineering of titanium-based implants., (Copyright 2007 Wiley Periodicals, Inc.)

The objective of this study was to surface modify the poly (D, L-lactic acid) (PDLLA) films and assess the effects of the modified surfaces on the functions of osteoblasts cultured in vitro. A layer-by-layer (LBL) self assembly technique, was used leading to the formation of multilayers on the PDLLA film surfaces. Chitosan (Chi) and poly (styrene sulfonate, sodium salt) (PSS) were utilized as polycation and polyanion in this study, respectively. The layer structure was investigated by using X-ray photoelectron spectroscopy (XPS) and water contact angle measurement, respectively. XPS analysis displayed the presence of chitosan on PDLLA surface. A full coverage of coating with PSS/Chi layers was achieved on the PDLLA surface only after the deposition layers of PEI/(PSS/Chi)2. These results showed that PDLLA films could be modified with PSS/Chi pairs which may affect the biocompatibility of the modified PDLLA films. To confirm this hypothesis, cell proliferation, cell viability as well as alkaline phosphtase activity of osteoblasts on layer-by-layer modified PDLLA films as well as control samples were investigated in vitro. The proliferation of osteoblasts on modified PDLLA films was found to be greater than that on control (p < 0.05 and p < 0.01) after 1, 4 and 7 days culture, respectively. Cell viability measurement showed that the PSS/Chi modified PDLLA films have higher cell viability (p < 0.01) than control. Osteoblast differentiation function (ALP) on LBL-modified PDLLA film was found significantly higher (p < 0.01) than that of virgin PDLLA films. These data suggests that PSS/Chi pair was successfully employed to surface modify PDLLA film via a layer-by-layer technique, and enhanced its cell biocompatibility.

One of the challenges in the field of tissue engineering is the development of biomaterial/cell interactions. For the purposes of the present study, two molecular weights of poly(aspartic acid) (PASP) were used to modify poly(D,L-lactic acid) (PDLLA) films in order to enhance their cell affinity. The properties of the PDLLA-modified surfaces and the controls were investigated by water contact angle measurement and electron spectroscopy for chemical analysis (ESCA). These data reflect the change in the biocompatibility of modified PDLLA surfaces. Then rat osteoblasts were seeded onto these modified surfaces and on controls to examine their effects on cell adhesion and proliferation. Cell morphologies on these surfaces were studied by scanning electron microscopy (SEM), and cell viability was evaluated with a MTT assay. In addition, differentiated cell function was assessed by measuring alkaline phosphatase (ALP) activity. The results suggest that PASP-modified surfaces may enhance the interactions between osteoblasts and PDLLA films., (Copyright 2002 Wiley Periodicals, Inc.)

Chitosan is a good biodegradable natural polymer, widely used in biomedical fields. In this study, chitosan was used to modify the surface of poly (D,L-lactic acid) (PDLLA) in order to enhance its cell affinity. The properties of a modified PDLLA surface and control were investigated by contact angle and electron spectroscopy for chemical analysis (ESCA), which indicated the changes in surface energy and chemical structure. Scanning electron microscopy (SEM) observation displayed differences in surface morphology between the chitosan-modified film and the control. These data reflected that PDLLA films could be modified with chitosan and in turn may affect the biocompatibility of the modified films. Therefore, adhesion and growth of osteoblasts on modified PDLLA films as well as control were studied. Cell morphologies on the films were examined by SEM and cell viability was evaluated using an MTT assay; the differentiated cell function was assessed by measuring alkaline phosphatase (ALP) activity. The ALP activity of modified PDLLA films was significantly higher than that found on the control (p < 0.01). The proliferation of osteoblasts on modified films was also found to be higher than that on the control (p < 0.05), suggesting that chitosan could be used to modify PDLLA and then enhance its cell biocompatibility., (Copyright 2002 Wiley Periodicals, Inc.)

The objective of this study was to investigate the efficiency of two treatments for poly(D,L-lactic acid) (PDLLA) surface modification using silk fibroin. one chemical treatment and one physical treatment: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (WSC) and entrapment. The properties of control films, WSC-modified and entrapment-treated PDLLA films were investigated by water contact angle measurement and electron spectroscopy for chemical analysis (ESCA). The water-contact angle measurement indicated the change of hydrophilicity and the ESCA analysis suggested that the modified PDLLA film using silk fibroin became enriched with nitrogen atoms. The biocompatibility of PDLLA film might be altered, which in turn would affect the functions of cells that were seeded on it. Therefore, attachment and proliferation of osteoblasts that were seeded on modified PDLLA films and control films were examined. Cell viability was evaluated by the MTT assay and differentiated cell function was assessed by measuring alkaline phosphatase activity. These results suggested that silk fibroin was used to modify PDLLA surface via WSC and that entrapment could improve the interactions between osteoblasts and PDLLA films. The entrapment treatment was more effective thin WSC treatment to accomplish the goal of surface modification.

The objective of this study was to modify the surface of poly(D,L-lactic acid) (PDLLA) with different molecular weight of silk fibroins, and assess the effects of the modified surfaces on the functions of rat osteoblasts cultured in vitro. The properties of the modified PDLLA surface and the control one were investigated by contact angle and electron spectroscopy for chemical analysis (ESCA). The former indicated the variation of hydrophilicity and the latter suggested that the modified PDLLA film using silk fibroin is enriched with nitrogen atoms. The biocompatibility of the PDLLA film may be altered and in turn affects the seeded cell functions. Therefore, attachment and proliferation of osteoblasts seeded on the modified PDLLA films and the control one were examined. Cell morphologies on these films were studied by scanning electron microscopy (SEM) and cell viability was evaluated by MTT assay. In addition, differentiated cell function was assessed by measuring the alkaline phosphatase (ALP) activity. These results suggest that the silk fibroin-modified PDLLA surface can improve the interaction between osteoblasts and the PDLLA films.

Poly(D,L-lactic acid) (PDLLA) was modified with alkylated chitosan (N-butyl chitosan and N-cetyl chitosan), and the effects of modified films on the functions of rat osteoblasts were investigated. The characteristics of surfaces (both modified and control) were examined by water contact angle measurement and electron spectroscopy for chemical analysis (ESCA). Cell morphologies on these surfaces were taken using scanning electron microscopy (SEM). Cell attachment and proliferation were used to assess cell behavior on modified surface and control. MTT assay was used to determined cell viability, and alkaline phosphatase (ALP) activity was taken to evaluate differentiated cell function. Compared with the untreated films, no significant difference in cell attachment of osteoblasts was found on the modified films at a period of 8 h (p > 0.05). However, cell proliferation of N-butyl chitosan rather than N-cetyl chitosan modified PDLLA films was significantly higher than that found on control one (p < 0.05) at the end of the 4th and 7th days. The cell viability of osteoblasts on N-butyl chitosan modified PDLLA films were found higher than that on control (p < 0.05). These results suggested that N-butyl chitosan contributed greater than N-cetyl chitosan when used to modify PDLLA films for improving its biocompatibility.

To treat the osteoporosis and regulate the biological behaviors of both osteoblasts and osteoclasts, we prepared a natural polysaccharide-derived nanocomposite, containing alendronate-grafted hyaluronate (HA-Aln) and bone morphogenetic protein 2 (BMP-2) and investigated its synergistic regulation on the behaviors of osteoblasts and osteoclastsin vitro. The HA-Aln/BMP-2 nanocomposite was fabricated through the electrostatic interactions between the HA-Aln molecule and BMP-2 molecule. Here, BMP-2 was used to improve the osteoblast-mediated bone formation. Alendronate (Aln), a targeting ligand to bone matrix, was used to inhibit the osteoclast-mediated bone resorption.In vitroresults showed that HA-Aln/BMP-2 nanocomposite could effectively maintain the bioactivity of loaded drugs. The osteoblasts that treated with the HA-Aln/BMP-2 nanocomposite presented a higher level of cell motility, alkaline phosphatase (ALP) activity, mineralization capacity, and osteoblast-related gene expressions (runt-related transcription factor 2, osterix, ALP, collagen type I, osteocalcin, and osteopontin), as compared to that of control group. Besides, the RAW264.7 cells that were treated with HA-Aln/BMP-2 nanocomposite showed a lower level of osteoclastic differentiation. Overall, the HA-Aln/BMP-2 nanocomposite exhibits promising potential as an efficient carrier for co-delivery of anti-osteoporotic drug and growth factors to promote osteoblastic differentiation and bone formation while suppressing osteoclastic activity. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • Ag nanoparticles loaded TiO2 nanotubes array was fabricated on Ti substrates. • Multilayered film coating was fabricated to seal TiO2 nanotubes array. • The present system achieved the controlled release of Ag ions. • The modified Ti substrates favored the growth of osteoblasts. • Antibacterial property of Ti substrates was significantly improved. Abstract In orthopedic fields, effective anti-infection property and promotive biocompatibility on surface of titanium implants are two crucial factors for long-term successful implants. Herein, Ag nanoparticles (NPs) loaded TiO 2 nanotubes (TNT) arrays were fabricated on Ti substrates with assistance of ultraviolet irradiation. Then, bioactive multilayer films of chitosan (CHI) and dialdehyde alginate (ADA) pair were deposited onto the Ag-loaded TNT arrays via a layer-by-layer (LBL) self-assembly technique, which could effectively achieve the impactful antibacterial ability of titanium and endow the substrates with favorable biocompatibility. The driving force of the assembling of multilayer films came from two sources, electrostatic interaction and covalent interaction of Schiff-bonds between CHI and ADA. The surface topography and wettability of different samples were characterized by field emission scanning electron microscopy, transmission electron microscopy and contact angle measurements, respectively. In addition, Ag ions release from TNT-Ag and LBL substrate was measured via inductively coupled plasma atomic emission spectroscopy (ICP-AES). The results of a series of biological behaviors of osteoblasts on different substrates in vitro , including lactate dehydrogenase activity assay, cytoskeleton observation and cell viability measurement, confirmed that LBL substrates coated with (ADA-CHI) 10 multilayer films have negligible cytotoxicity and promote osteoblast growth compared with TNT-Ag substrates, which could ascribe to the slow-release of Ag ions and the biocompatibility of (ADA-CHI) 10 multilayer. More importantly, owing to the release of Ag ions, the LBL samples still exhibited a prominent antibacterial activity for S.aureus and E.coli. Characteristics of bacterial adhesion and viability measurement proved that the fabricated Ag-incorporated platform was capable of obviously inhibiting the adhesion and growth of bacteria. Therefore, this approach of surface modification for Ti substrates presented here may provide an alternative strategy to simultaneously meet the desirable osteoblast growth and reduce bacterial infection for implants in clinical application. [ABSTRACT FROM AUTHOR]

The insufficient osseointegration and bacterial infection of titanium and its alloys remain the key challenges in their clinic applications, which may result in failure implantation. To improve osteogenetic and antibacterial properties, TiO 2 nanotube arrays were fabricated on titanium substrates for loading of antibacterial drug. Then, TiO 2 nanotube arrays were covered with chitosan/sodium alginate multilayer films. The successful construction of this system was verified via scanning electron microscopy and contact angle measurement. The cytocompatibility evaluation in vitro , including cytoskeleton observation, cell viability measurement, and alkaline phosphatase activity assay, confirmed that the present system was capable of accelerating the growth of osteoblasts. In addition, bacterial adhesion and viability assay verified that treated Ti substrates were capable of reducing the adhesion of bacteria. This study may provide an alternative to develop titanium-based implants for enhanced bone osseointegration and reduced bacterial infection. [ABSTRACT FROM AUTHOR]

Bacterial infection is one of the most severe postoperative complications leading to implantation failure. The early bacterial stage (4–6 h) was proved to be the “decisive period” for long-term bacteria-related infection. Thus, to endow potential early antibacterial capacity for a titanium (Ti) based implant, an effective antiseptic agent of octenidine dihydrochloride (OCT) was effectively loaded on the mesoporous silica nanoparticles (MSNs)-incorporated titania coating which was fabricated by an electrophoretic-enhanced micro-arc oxidation technique. The surface characteristic of the coatings were characterized by various methods (SEM, AFM, XPS, XRD, etc.), and its corrosion resistance was also examined by the potentiodynamic polarization curves. The composite coating without OCT loading not only displayed good cytocompatibility but also exhibited certain anti-bacterial property. After loading with OCT, its antibacterial efficiency of the titanium substrates with composite coating was greatly enhanced without compromising their cytocompatibility. The study provides an approach for the fabrication of anti-bacterial Ti implant for potential orthopedic application. [ABSTRACT FROM AUTHOR]

Due to its good biocompatibility and chemical stability, tantalum attracts much attention for biomedical applications, however, limited by its relatively high cost. In this study, tantalum oxide layers were deposited onto titanium (Ti) substrates with polymer-assisted deposition (PAD) technique to improve their corrosion resistance and cytocompatibility. Scanning electron microscopy (SEM), atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS), X-ray diffraction (XRD), scratch tests and water contact angle measurement proved the successful formation of tantalum oxide layers onto the surfaces of Ti substrates. The potentiodynamic polarization curves revealed that the formed tantalum oxide layers improved the corrosion resistance of Ti substrates. Moreover, the in vitro tests displayed that Ti substrates coated with 6 layers of tantalum oxide promoted proliferation, alkaline phosphatase activity, mineralization and osteogenic gene expressions (ALP, Col I, OC and OPN) of osteoblasts, respectively. The study affords an approach for the fabrication of Ti-based implant with improved corrosion resistance and cytocompatibility. [ABSTRACT FROM AUTHOR]

Highlights: [•] Titanium substrates were surface functionalized with cecropin B via dopamine. [•] Cecropin B immobilized titanium substrates promoted the proliferation of osteoblasts. [•] The surface functionalized titanium substrates displayed antibacterial property. [•] Cecropin B immobilized titanium substrates reduced the inflammation responses of macrophages. [Copyright &y& Elsevier]