Effect Of Nanotopography Research Articles

Probing Nanotopography-Mediated Macrophage Polarization via Integrated Machine Learning and Combinatorial Biophysical Cue Mapping.

Inflammatory responses, leading to fibrosis and potential host rejection, significantly hinder the long-term success and widespread adoption of biomedical implants. The ability to control and investigated macrophage inflammatory responses at the implant-macrophage interface would be critical for reducing chronic inflammation and improving tissue integration. Nonetheless, the systematic investigation of how surface topography affects macrophage polarization is typically complicated by the restricted complexity of accessible nanostructures, difficulties in achieving exact control, and biased preselection of experimental parameters. In response to these problems, we developed a large-scale, high-content combinatorial biophysical cue (CBC) array for enabling high-throughput screening (HTS) of the effects of nanotopography on macrophage polarization and subsequent inflammatory processes. Our CBC array, created utilizing the dynamic laser interference lithography (DLIL) technology, contains over 1 million nanotopographies, ranging from nanolines and nanogrids to intricate hierarchical structures with dimensions ranging from 100 nm to several microns. Using machine learning (ML) based on the Gaussian process regression algorithm, we successfully identified certain topographical signals that either repress (pro-M2) or stimulate (pro-M1) macrophage polarization. The upscaling of these nanotopographies for further examination has shown mechanisms such as cytoskeletal remodeling and ROCK-dependent epigenetic activation to be critical to the mechanotransduction pathways regulating macrophage fate. Thus, we have also developed a platform combining advanced DLIL nanofabrication techniques, HTS, ML-driven prediction of nanobio interactions, and mechanotransduction pathway evaluation. In short, our developed platform technology not only improves our ability to investigate and understand nanotopography-regulated macrophage inflammatory responses but also holds great potential for guiding the design of nanostructured coatings for therapeutic biomaterials and biomedical implants.

Effects of material nano-topography on the angiogenesis of type H vessels: Size dependence, cell heterogeneity and intercellular communication

Type H vessel, a vascular subtype in bone, is a critical regulator of osteogenesis, but how material properties affect this organ-specific vessel remains unknown. Here, titania nanotubes were fabricated on bone implant surface to investigate the effects of nano-topography on type H vessels. In vivo, surface nanotubes with 20–100 nm diameters promoted the angiogenesis of type H vessels and bone regeneration in mouse femurs to different extents, with the best effects induced by 70 nm diameter. In vitro, bone-specific endothelial cells (BECs) and artery endothelial cells (AECs) presented significantly different behaviors on the same material. Nanotubes with 20 nm small diameters significantly improved the adhesion, proliferation, type H differentiation of BECs and their paracrine function to regulate pre-osteoblasts (POBs), possibly via binding integrin β1 on the cell membrane, but these effects weakened when tube diameter increased, which conflicted with the results in vivo. Further study suggested that the better in vivo effects by larger diameters of 70–100 nm might be exerted indirectly through remodeling the regulation from POBs to BECs, highlighting the underappreciated indirect bio-effects of materials via intercellular communication. These suggest that nanoscale material topography makes significant impact on the angiogenesis of type H vessels, directly via binding integrins on the cell membrane of BECs and indirectly via modulating the regulation from osteoblastic cells to BECs, both in a size-dependent manner. Cells of the same type but from different tissues may show different responses to the same material, thus material properties should be tailored to the specific cell population. In research on material-tissue interactions, conclusions from in vitro experiments exposing a single type of cell to material might deviate from the truth in vivo, because materials may indirectly influence the targeted cells through modulating intercellular communication. These provide new insights into material-tissue interactions.

Nanotopography and Microconfinement Impact on Primary Hippocampal Astrocyte Morphology, Cytoskeleton and Spontaneous Calcium Wave Signalling.

Astrocytes' organisation affects the functioning and the fine morphology of the brain, both in physiological and pathological contexts. Although many aspects of their role have been characterised, their complex functions remain, to a certain extent, unclear with respect to their contribution to brain cell communication. Here, we studied the effects of nanotopography and microconfinement on primary hippocampal rat astrocytes. For this purpose, we fabricated nanostructured zirconia surfaces as homogenous substrates and as micrometric patterns, the latter produced by a combination of an additive nanofabrication and micropatterning technique. These engineered substrates reproduce both nanotopographical features and microscale geometries that astrocytes encounter in their natural environment, such as basement membrane topography, as well as blood vessels and axonal fibre topology. The impact of restrictive adhesion manifests in the modulation of several cellular properties of single cells (morphological and actin cytoskeletal changes) and the network organisation and functioning. Calcium wave signalling was observed only in astrocytes grown in confined geometries, with an activity enhancement in cells forming elongated agglomerates with dimensions typical of blood vessels or axon fibres. Our results suggest that calcium oscillation and wave propagation are closely related to astrocytic morphology and actin cytoskeleton organisation.

A biomimetic hyaluronic acid-silk fibroin nanofiber scaffold promoting regeneration of transected urothelium.

This study was designed to investigate the regulatory effect of hyaluronic acid (HA)—coating silk fibroin (SF) nanofibers during epithelialization of urinary tract for urethral regeneration. The obtained electrospun biomimetic tubular HA‐SF nanofiber scaffold is composed of a dense inner layer and a porous outer layer in order to mimic adhesion and cavernous layers of the native tissue, respectively. A thin layer of HA‐gel coating was fixed in the inner wall to provide SF nanofibers with a dense and smooth surface nano‐topography and higher hydrophilicity. Compared with pure SF nanofibers, HA‐SF nanofibers significantly promoted the adhesion, growth, and proliferation of primary urothelial cells, and up‐regulate the expression of uroplakin‐3 (terminal differentiation keratin protein in urothelium). Using the New Zealand male rabbit urethral injury model, the scaffold composed of tubular HA‐SF nanofibers could recruit lumen and myoepithelial cells from the adjacent area of the host, rapidly reconstructing the urothelial barrier in the wound area in order to keep the urinary tract unobstructed, thereby promoting luminal epithelialization, smooth muscle bundle structural remodeling, and capillary formation. Overall, the synergistic effects of nano‐topography and biophysical cues in a biomimetic scaffold design for effective endogenous regeneration.

Influence of Muscovite (001) Surface Nanotopography on Radionuclide Adsorption Studied by Kinetic Monte Carlo Simulations

Mechanistic understanding and prediction of solute adsorption from fluids onto mineral surfaces is relevant for many natural and technical processes. Mineral surfaces in natural systems are often exposed to fluids at non-equilibrium conditions resulting in surface dissolution reactions. Such reactions cause the formation of surface nanotopography and, consequently, the exposure of different types of surface atoms. The quantitative effect of nanotopography on the efficiency of adsorption reactions at crystal surfaces is not known. Using kinetic Monte Carlo simulations, we combined a model of muscovite (001) face dissolution with a consequent model of radionuclide adsorption on the rough mineral surface. The model considers three different adsorption sites based on the muscovite surface cations: silicon, tetrahedral, and octahedral aluminum. Two different nanotopography configurations are investigated, both showing similar adsorption behavior. Octahedral aluminum surface atoms defined by having the highest reactivity toward adsorption are exposed solely on steps and pits on the muscovite (001) face. Thus, their availability directly depends on the surface nanotopography. The model results show the need for a more precise parameterization of surface site-specific adsorption, taking into account the coordination of the involved surface cation such as kink, step, or terrace sites.

Surface engineering of biomaterials in orthopedic and dental implants: Strategies to improve osteointegration, bacteriostatic and bactericidal activities

The success of biomedical implants in orthopedic and dental applications is usually limited due to insufficient bone-implant integration, and implant-related infections. Biointerfaces are critical in regulating their interactions and the desirable performance of biomaterials in biological environment. Surface engineering has been widely studied to realize better control of the interface interaction to further enhance the desired behavior of biomaterials. This review aims to investigate surface coating strategies in hard tissue applications to address insufficient osteointegration and implant-related infection problems. We first focused on surface coatings to enhance the osteointegration and biocompatibility of implants by emphasizing calcium phosphate-related, nanoscale TiO2 -related, bioactive tantalum-based and biomolecules incorporated coatings. Different coating strategies such as plasma spraying, biomimetic deposition, electrochemical anodization and LENS are discussed. We then discussed techniques to construct anti-adhesive and bactericidal surface while emphasizing multifunctional surface coating techniques that combine potential osteointegration and antibacterial activities. The effects of nanotopography via TiO2 coatings on antibacterial performance are interesting and included. A smart bacteria-responsive titanium dioxide nanotubes coating is also attractive and elaborated. Developing multifunctional surface coatings combining osteogenesis and antimicrobial activity is the current trend. Surface engineering methods are usually combined to obtain hierarchical multiscale surface structures with better biofunctionalization outcomes.

New perspectives on the roles of nanoscale surface topography in modulating intracellular signaling

The physical properties of biomaterials, such as elasticity, stiffness, and surface nanotopography, are mechanical cues that regulate a broad spectrum of cell behaviors, including migration, differentiation, proliferation, and reprogramming. Among them, nanoscale surface topography, i.e., nanotopography, defines the nanoscale shape and spatial arrangement of surface elements, which directly interact with the cell membranes and stimulate changes in the cell signaling pathways. In biological systems, the effects of nanotopography are often entangled with those of other mechanical and biochemical factors. Precise engineering of 2D nanopatterns and 3D nanostructures with well-defined features has provided a powerful means to study the cellular responses to specific topographic features. In this Review, we discuss efforts in the last three years to understand how nanotopography affects membrane receptor activation, curvature-induced cell signaling, and stem cell differentiation.

Polycaprolactone/Gelatin/Hyaluronic Acid Electrospun Scaffolds to Mimic Glioblastoma Extracellular Matrix.

Glioblastoma (GBM), one of the most malignant types of human brain tumor, is resistant to conventional treatments and is associated with poor survival. Since the 3D extracellular matrix (ECM) of GBM microenvironment plays a significant role on the tumor behavior, the engineering of the ECM will help us to get more information on the tumor behavior and to define novel therapeutic strategies. In this study, polycaprolactone (PCL)/gelatin(Gel)/hyaluronic acid(HA) composite scaffolds with aligned and randomly oriented nanofibers were successfully fabricated by electrospinning for mimicking the extracellular matrix of GBM tumor. We investigated the effect of nanotopography and components of fibers on the mechanical, morphological, and hydrophilic properties of electrospun nanofiber as well as their biocompatibility properties. Fourier transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC) have been used to investigate possible interactions between components. The mean fiber diameter in the nanofiber matrix was increased with the presence of HA at low collector rotation speed. Moreover, the rotational velocity of the collector affected the fiber diameters as well as their homogenous distribution. Water contact angle measurements confirmed that hyaluronic acid-incorporated aligned nanofibers were more hydrophilic than that of random nanofibers. In addition, PCL/Gel/HA nanofibrous scaffold (7.9 MPa) exhibited a significant decrease in tensile strength compared to PCL/Gel nanofibrous mat (19.2 MPa). In-vitro biocompatibilities of nanofiber scaffolds were tested with glioblastoma cells (U251), and the PCL/Gel/HA scaffolds with random nanofiber showed improved cell adhesion and proliferation. On the other hand, PCL/Gel/HA scaffolds with aligned nanofiber were found suitable for enhancing axon growth and elongation supporting intracellular communication. Based on these results, PCL/Gel/HA composite scaffolds are excellent candidates as a biomimetic matrix for GBM and the study of the tumor.

Effects of Nanotopography Regulation and Silicon Doping on Angiogenic and Osteogenic Activities of Hydroxyapatite Coating on Titanium Implant.

BackgroundAngiogenic and osteogenic activities are two major problems with biomedical titanium (Ti) and other orthopedic implants used to repair large bone defects.PurposeThe aim of this study is to prepare hydroxyapatite (HA) coatings on the surface of Ti by using electrochemical deposition (ED), and to evaluate the effects of nanotopography and silicon (Si) doping on the angiogenic and osteogenic activities of the coating in vitro.Materials and MethodsHA coating and Si-doped HA (HS) coatings with varying nanotopographies were fabricated using two ED modes, ie, the pulsive current (PC) and cyclic voltammetry (CV) methods. The coatings were characterized through scanning electron microscope (SEM), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), X-ray photoelectron spectrometer (XPS), and atomic force microscopy (AFM), and their in vitro bioactivity and protein adsorption were assessed. Using MC3T3-E1 pre-osteoblasts and HUVECs as cell models, the osteogenic and angiogenic capabilities of the coatings were evaluated through in vitro cellular experiments.ResultsBy controlling Si content in ~0.8 wt.%, the coatings resulting from the PC mode (HA-PC and HS-PC) and CV mode (HA-CV and HS-CV) had nanosheet and nanorod topographies, respectively. At lower crystallinity, higher ionic dissolution, smaller contact angle, higher surface roughness, and more negative zeta potential, the HS and PC samples exhibited quicker apatite deposition and higher BSA adsorption capacity. The in vitro cell study showed that Si doping was more favorable for enhancing the viability of the MC3T3-E1 cells, but nanosheet coating increased the area for cell spreading. Of the four coatings, HS-PC with Si doping and nanosheet topography exhibited the best effect in terms of up-regulating the expressions of the osteogenic genes (ALP, Col-I, OSX, OPN and OCN) in the MC3T3-E1 cells. Moreover, all leach liquors of the surface-coated Ti disks promoted the growth of the HUVECs, and the HS samples played a more significant role in promoting cell migration and tube formation than the HA samples. Of the four leach liquors, only the two HS samples up-regulated NO content and expressions of the angiogenesis-related genes (VEGF, bFGF and eNOS) in the HUVECs, and the HS-PC yielded a better effect.ConclusionThe results show that Si doping while regulating the topography of the coating can help enhance the bone regeneration and vascularization of HA-coated Ti implants.

Predicting gene expression using morphological cell responses to nanotopography

Cells respond in complex ways to their environment, making it challenging to predict a direct relationship between the two. A key problem is the lack of informative representations of parameters that translate directly into biological function. Here we present a platform to relate the effects of cell morphology to gene expression induced by nanotopography. This platform utilizes the ‘morphome’, a multivariate dataset of cell morphology parameters. We create a Bayesian linear regression model that uses the morphome to robustly predict changes in bone, cartilage, muscle and fibrous gene expression induced by nanotopography. Furthermore, through this model we effectively predict nanotopography-induced gene expression from a complex co-culture microenvironment. The information from the morphome uncovers previously unknown effects of nanotopography on altering cell–cell interaction and osteogenic gene expression at the single cell level. The predictive relationship between morphology and gene expression arising from cell-material interaction shows promise for exploration of new topographies.

Effect of silica nano-spheres on adhesion of oral bacteria and human fibroblasts

Objective This study investigated the effect of surface nano-patterning on adhesion of an oral early commensal colonizer, Streptococcus mitis and the opportunistic pathogen Staphylococcus aureus and human fibroblasts (HDFa) in a laminar flow cell. Methods Nanostructured surfaces were made by functionalizing glass substrates with 40 nm SiO2 nanoparticles. Gradients in nanoparticle surface coverage were fabricated to study the effect of nanoparticle spacing within a single experiment. Bacterial adhesion was investigated after 5 min of contact time by subjecting surfaces to a flow in a laminar flow cell. In addition, to examine the particles effect on human cells, the establishment of focal adhesion and spreading of primary human dermal fibroblasts (HDFa) were investigated after 4 and 24 h. Results Adhesion of both S. aureus and S. mitis decreased on surfaces functionalized with nanoparticles and coincided with higher nanoparticle surface coverage on the surface. Both strains were tested on three separate surfaces. The regression analysis showed that S. mitis was influenced more by surface modification than S. aureus. The establishment of focal adhesions in HDFa cells was delayed on the nanostructured part of the surfaces after both 4 and 24 h of culturing. Significance In the current manuscript, we have used a flow cell to investigate the effect of nanotopographies on S. aureus and S. mitis adhesion. The present findings are of relevance for design of future implant and prostheses surfaces in order to reduce adhesion of bacteria.

Regulating response and leukocyte adhesion of human endothelial cell by gradient nanohole substrate

Understanding signals in the microenvironment that regulate endothelial cell behavior are important in tissue engineering. Although many studies have examined the cellular effects of nanotopography, no study has investigated the functional regulation of human endothelial cells grown on nano-sized gradient hole substrate. We examined the cellular response of human umbilical vein endothelial cells (HUVECs) by using a gradient nanohole substrate (GHS) with three different types of nanohole patterns (HP): which diameters were described in HP1, 120–200 nm; HP2, 200–280 nm; HP3, 280–360 nm. In results, HP2 GHS increased the attachment and proliferation of HUVECs. Also, gene expression of focal adhesion markers in HUVECs was significantly increased on HP2 GHS. In vitro tube formation assay showed the enhancement of tubular network formation of HUVECs after priming on GHS compared to Flat. Furthermore, leukocyte adhesion was also reduced in the HUVECs in a hole-diameter dependent manner. To summarize, optimal proliferations with reduced leukocyte adhesion of HUVECs were achieved by gradient nanohole substrate with 200–280 nm-sized holes.

Abstract 17236: Regulating Optimal Angiogenesis and Leukocyte Adhesion of Human Endothelial Cells by Gradient Nanopore Substrate

Introduction: Understanding signals in the microenvironment that regulate endothelial cell behavior are important in tissue engineering. Hypothesis: Although many studies have examined the cellular effects of nanotopography, no study has investigated the functional regulation of human endothelial cells grown on nano-sized gradient pore substrate. Methods: We examined the cellular response of human umbilical vein endothelial cells (HUVECs) by using a gradient nanopore substrate (GPS) with three different types of hole nanopore patterns (HP): which diameters were described in HP1, 120-200 nm; HP2, 200-280 nm; HP3, 280-360 nm. Results: Our study revealed that: (1) HUVECs showed highest proliferation and attachment capacity on HP2 (200-280 nm) GPS, (2) HP2 GPS was determined to be the optimal diameter for extending greater angiogenic potential to HUVECs and (3) both HP2 and HP3 GPS significantly decrease leukocyte adhesion compared to Flat. Moreover, expressions of VCL, TLN1, PXN, ITGA2, ITGA6, ITGAV, ITGB3 and ROCK1 in HUVECs cultured on HP2 GPS were significantly greater than that of Flat. Conversely, TLN1, PXN, ROCK 1 and ROCK2 mRNA levels decreased on HP3 GPS compared to those of Flat. Mature tube structures at 24 hours were found in HUVECs stimulated by HP2 GPS. Expressions of angiogenesis related markers (ANGPT1, TIE2 and CXCR4) and endothelial-related marker (vWF) were significantly increased in HUVECs grown on HP2 GPS. The adhesion of peripheral blood mononuclear cells (PBMNCs) to HUVECs stimulated by HP2 and HP3 GPS was significantly reduced compared to those of Flat at 3, 6 and 24 hours after PBMNC seeding. Conclusions: Optimal functional regulations of HUVECs were achieved by nanopore substrate with 200-280 nm-sized pores. Our results provide the possibility of applying nanopore interface-based implant devices in the field of cardiovascular regenerative therapy.

Nanoengineered, cell-derived extracellular matrix influences ECM-related gene expression of mesenchymal stem cells

BackgroundHuman mesenchymal stem cells (hMSCs) are, due to their pluripotency, useful sources of cells for stem cell therapy and tissue regeneration. The phenotypes of hMSCs are strongly influenced by their microenvironment, in particular the extracellular matrix (ECM), the composition and structure of which are important in regulating stem cell fate. In reciprocal manner, the properties of ECM are remodeled by the hMSCs, but the mechanism involved in ECM remodeling by hMSCs under topographical stimulus is unclear. In this study, we therefore examined the effect of nanotopography on the expression of ECM proteins by hMSCs by analyzing the quantity and structure of the ECM on a nanogrooved surface.MethodsTo develop the nanoengineered, hMSC-derived ECM, we fabricated the nanogrooves on a coverglass using a UV-curable polyurethane acrylate (PUA). Then, hMSCs were cultivated on the nanogrooves, and the cells at the full confluency were decellularized. To analyze the effect of nanotopography on the hMSCs, the hMSCs were re-seeded on the nanoengineered, hMSC-derived ECM.ResultshMSCs cultured within the nano-engineered hMSC-derived ECM sheet showed a different pattern of expression of ECM proteins from those cultured on ECM-free, nanogrooved surface. Moreover, hMSCs on the nano-engineered ECM sheet had a shorter vinculin length and were less well-aligned than those on the other surface. In addition, the expression pattern of ECM-related genes by hMSCs on the nanoengineered ECM sheet was altered. Interestingly, the expression of genes for osteogenesis-related ECM proteins was downregulated, while that of genes for chondrogenesis-related ECM proteins was upregulated, on the nanoengineered ECM sheet.ConclusionsThe nanoengineered ECM influenced the phenotypic features of hMSCs, and that hMSCs can remodel their ECM microenvironment in the presence of a nanostructured ECM to guide differentiation into a specific lineage.

Temporal Changes in Nucleus Morphology, Lamin A/C and Histone Methylation During Nanotopography-Induced Neuronal Differentiation of Stem Cells.

Stem cell differentiation can be regulated by biophysical cues such as nanotopography. It involves sensing and integration of these biophysical cues into their transcriptome with a mechanism that is yet to be discovered. In addition to the cytoskeletal and focal adhesion remodeling, nanotopography has also been shown to modulate nucleus morphology. Here, we studied the effect of nanotopography on the temporal changes in nuclei of human embryonic stem cells (hESCs) and human mesenchymal stem cells (hMSCs). Using a high throughput Multi-architecture (MARC) chip analysis, the circularity of the stem cell nuclei changed significantly on different patterns. Human ESCs and MSCs showed different temporal changes in nucleus morphology, lamin A/C expression and histone methylation during topography-induced neuronal differentiation. In hESCs, the expression of nuclear matrix protein, lamin A/C during neuronal differentiation of hESCs on PDMS samples were weakly detected in the first 7 days of differentiation. The histone 3 trimethylation on Lysine 9 (H3K9me3) decreased after differentiation initiated and showed temporal changes in their expression and organization during neuronal differentiation. In hMSCs, the expression of lamin A/C was significantly increased after the first 24 h of cell culture. The quantitative analysis of histone methylation also showed a significant increase in hMSCs histone methylation on 250 nm anisotropic nanogratings within the first 24 h of seeding. This reiterates the importance of cell-substrate sensing within the first 24 h for adult stem cells. The lamin A/C expression and histone methylation shows a correlation of epigenetic changes in early events of differentiation, giving an insight on how extracellular nanotopographical cues are transduced into nuclear biochemical signals. Collectively, these results provide more understanding into the nuclear regulation of the mechanotransduction of nanotopographical cues in stem cell differentiation.

Curvature-dependent effects of nanotopography on classical immune complement activation

The aim of this study was to investigate how the size of nanosized surface features affect classical immune complement activation through adsorption of IgG and the following binding of C1q. By using model surfaces with immobilized SiO2 nanoparticles of different sizes (8, 32 and 68 nm in diameter), three different curvatures with the same chemistry was systematically studied and analyzed using the acoustic sensing technique; Quartz Crystal Microbalance with Dissipation Monitoring (QCM-D). Circular Dichroism (CD) was employed to study any changes in the secondary structure of IgG using a methodology with stacked functionalized substrates. Our results show that the amount of IgG adsorption increased slightly with nanoparticle size, but also showed a strong size/curvature-dependent effect on the following C1q binding, with the highest binding to IgG adsorbed on the largest nanoparticles and a smooth control surface, indicating that classical immune complement activation possibly increase with decreasing curvature. We conclude that the difference in C1q binding was not due to changes in the secondary structure of IgG, suggesting that geometrical arrangement of adsorbed IgG is the determining factor. Statement of SignificanceWe have shown that small changes at the topographical nanoscale can give large effects on the initiation of the classical immune complement cascade, an important immunological reaction that take place when a foreign material is inserted in the body. By developing a methodology using silicon dioxide nanoparticles with three different sizes, to systematically study their impact on the secondary structure and binding of human immunoglobulin G (IgG) to the initiator protein C1q of the classical complement cascade, we have shown that the initiation of the classical immune complement is hampered by the sharp curvature of the smaller nanoparticles. We conclude that this is not mediated by changes in the secondary structure of the adsorbed proteins, but rather an effect of curvature-induced spatial mismatch. The results provide a possible mechanistic explanation on how nanotopography may effect protein adsorption and protein cascade events.

Nanostructured Surface Modification to Bone Implants for Bone Regeneration.

Bone defect repair is a complex process of bone regeneration. Bone substitute implanted at the bone defect sites act as three-dimensional scaffolds that guide and promote bone regeneration. The function of orthopedic implants is often limited by complicating factors such as insufficient integration into host tissue, inflammatory reactions, and infection. Biological behaviors of immune cells and osteogenesis-related cells on these bone biomaterials are largely determined by the surface properties. The advances in nanofabrication technology have help us fabricated a range of nanostructured surface with controlled physicochemical properties to modulate the behaviors of osteogenesis-related cells and immune cells, thus affecting bone integration and local immune reaction. In this review, we summary the development of nanostructured surface modification to bone implants with controlled physicochemical properties, including roughness, wettability, surface charge and topography. In particular, we focus on the effect of nanotopographies on cell recruitment and cellular adhesion, osteogenic differentiation, mineralization and osseointegration, antibacterial activities, and modulation of immune response.

Gold nano-decorated aligned polyurethane nanofibers for enhancement of neurite outgrowth and elongation.

Neurite outgrowth and elongation of neural cells is the most important subject that is considered in nerve tissue engineering. In this regard, aligned nanofibers have taken much attention in terms of providing guidance for newly outgrown neurites. The main objective of this study was to fabricate aligned polyurethane nanofibers by electrospinning process and decorate them with gold nanoparticles to further investigate the synergistic effects of nanotopography, biological nerve growth factor (NGF) and electrical stimulations on neurite outgrowth and elongation of pheochromocytoma (PC-12) model cells. In this regard, smooth and uniform aligned polyurethane nanofibers with the average diameter of 519 ± 56 nm were fabricated and decorated with the gold nanoparticles with the average diameter of ∼50 nm. PC-12 cells were cultured on the various nanofiber surfaces inside the bio-mimetic bioreactor system and exposed either to NGF alone or combination of NGF and electrical stimulation. It was found that 50 ng/mL NGF concentration is an optimal value for the stimulation of neurite outgrowth. After 4 days of culture under 100 mV, 10 ms electrical stimulation in 1 h/day period it was found that the gold nanoparticle decorated aligned polyurethane nanofibers increased the neurite outgrowth and elongation more with the combinational NGF and electrical stimulation. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1604-1613, 2018.

Fabrication of Plant Virus-Based Thin Films to Modulate the Osteogenic Differentiation of Mesenchymal Stem Cells.

Stem cells can interact and respond to the extracellular nanoscale environment. Viral nanoparticles have been utilized as building blocks to control cell growth and differentiation. By integrating stem cell research and virus nanoparticle chemistry together, a systematic analysis of the effects of nanotopography on stem cell differentiation can be accomplished. The fabrication of thin films of the viral nanoparticles is particularly valuable for such studies. Here, we describe two methods to fabricate plant virus-based thin films and procedures to study the osteogenic differentiation of mesenchymal stem cells on plant virus-based substrates. The method makes use of wild-type tobacco mosaic virus (wt-TMV), RGD-modified TMV (TMV-RGD), turnip yellow mosaic virus (TYMV), cowpea mosaic virus (CPMV), turnip vein clearing virus (TVCV), and potato virus X (PVX) for development of bone tissue engineering biomaterials.

Aligned Fibers Fabricated by Near-Field Electrospinning Influence the Orientation and Differentiation of hPDLSCs for Periodontal Regeneration.

The periodontal ligament (PDL) is a specific connective tissue composed of organized and aligned collagenous fibers that fix the tooth root in the alveolar bone. The alignment of PDL fibers and their function have been neglected in the past, as many studies investigated the regeneration of the periodontium, including alveolar bone and cementum regeneration. In this study, we fabricated biodegradable aligned fibers (ε-caprolactone/collagen) by near-field electrospinning (NFE) to control the arrangement of human periodontal ligament stem cells (hPDLSCs), aiming to guide the oriented regeneration of the periodontal ligament. Compared with random electrospun fibers, the in vitro study investigated the effects of nanotopography on stem cell differentiation of hPDLSCs. The hPDLSCs were identified by flow cytometry, and the multipotency of hPDLSCs was confirmed by successful osteogenic induction. The hPDLSCs were co-cultured with aligned and random fibers. The cell morphology was observed by confocal micrograph and scanning electron micrograph, which showed that aligned fibers could guide the orientation and elongation of hPDLSCs. The expression of periodontal ligament-related genes was higher when cultured with aligned fibers than when cultured with random fibers. In conclusion, via near-field electrospinning, aligned biodegradable fibers were prepared and guided the orientation arrangement of hPDLSCs, providing a better microenvironment for periodontal ligament regeneration. This technology might be further used in the regeneration of tissues in a given direction.