Osteoblast Attachment Research Articles

A Synthetic Biodegradable Polymer Membrane for Guided Bone Regeneration in Bone Defect.

Guided bone regeneration (GBR) technique is most commonly used to treat alveolar bone defect. Polylactic acid (PLA) attracts much attention to utilize as a GBR membrane because it has relatively high mechanical strength and biodegradability. However, randomized controlled trials of PLA as a GBR membrane in animals were rare. The aim of this work is to observe the efficacy of polylactic acid membrane in guiding bone regeneration in Beagle canine alveolar bone defect restoration and to compare efficacy with the collagen membrane, providing an experimental basis for further clinical use of the polylactic acid membrane. The tests of physical and chemical properties showed that the PLA membrane has well mechanical strength to maintenance the space for the new bone, and has proper aperture for the attachment of osteoblasts. Through X-ray and histopathological examination of the different time points, the bone grafting material covered with PLA membrane can form similar mature bone compared to collagen membrane ones. Meanwhile, biodegradable speed of the PLA membrane was slower. Thus, this study showed that polylactic acid membrane as synthetic biodegradable polymer was reliably effective in guiding bone regeneration of alveolar bone defects, showed the favorable osteogenic capability and forecasts well applications in bone augmentation.

Strong, elastic, and tough high internal phase emulsions stabilized solely by cod myofibers for multidisciplinary applications

High internal phase emulsions (HIPEs) are versatile platforms for various applications owing to the low-density, interconnected structure, and large specific area. The mechanical properties of existing HIPEs, especially those stabilized solely by one naturally-occurring substance, hinder their practical applications. In this study, we disclose strong (up to 1.6 MPa), tough (up to 0.7 MJ m−3), and fatigue-resistant HIPEs stabilized solely by cod myofibers (CMs). Unlike conventional Pickering-type HIPEs, intrinsic strengthening and toughening of CMs, resultant from cadmium bridging of actin and myosin, endow the superior mechanical properties of our HIPEs. The HIPEs are conformable to 3D printing, yielding architectures with tailorable rheological properties that are vital for texture of foods. Besides, the network structures of our HIPEs serve the template of aerogels that had the oil absorption capacity of up to 58 g g−1 and maintained properties after repeated uses. The ability of aerogels to absorb petroleum or soy oil spilled on down feathers within 40 s shows the potential applications in fabric cleansing and remediation of oil spillage including rescuing seabirds. Furthermore, the porous structures of aerogels are excellent platforms for osteoblast attachment and proliferation and therefore are potential scaffolds in tissue engineering. The mechanically robust, reusable, and biofriendly HIPEs in this study can be developed as facile and reusable materials for numerous multidisciplinary applications.

Cellular Response to Sol-Gel Hybrid Materials Releasing Boron and Calcium Ions.

Poly(dimethylsiloxane) (PDMS)-SiO2-CaO-based hybrid materials prepared by sol-gel have proved to be very promising materials for tissue engineering applications and drug-delivery systems. These hybrids are biocompatible and present osteogenic and bioactive properties supporting osteoblast attachment and bone growth. The incorporation of therapeutic elements in these materials, such as boron (B) and calcium (Ca), was considered in this study as an approach to develop biomaterials capable of stimulating bone regeneration. The main purpose of this work was thus to produce, by sol-gel, bioactive and biocompatible hybrid materials of the PDMS-SiO2-B2O3-CaO system, capable of a controlled Ca and B release. Different compositions with different boron amounts were prepared using the same precursors resulting in different monolithic materials, with distinct structures and microstructures. Structural features were assessed by Fourier transform infrared (FT-IR) spectrometry and solid-state nuclear magnetic resonance (NMR) techniques, which confirmed the presence of hybrid bonds (Si-O-Si) between organic (PDMS) and inorganic phase (tetraethyl orthosilicate (TEOS)), as well as borosiloxane bonds (B-O-Si). From the 11B NMR results, it was found that Ca changes the boron coordination, from trigonal (BO3) to tetrahedral (BO4). Scanning electron microscopy (SEM) micrographs and N2 isotherms showed that the incorporation of boron modifies the material's microstructure by increasing the macroporosity and decreasing the specific surface area (SSA). In vitro tests in simulated body fluid (SBF) showed the precipitation of a calcium phosphate layer on the material surface and the controlled release of therapeutic ions. The cytocompatibility of the prepared hybrids was studied with bone marrow stromal cells (ST-2 cell line) by analyzing the cell viability and cell density. The results demonstrated that increasing the dilution rate of extraction medium from the hybrids leads to improved cell behavior. The relationship between the in vitro response and the structural and microstructural features of the materials was explored. It was shown that the release of calcium and boron ions, determined by the hybrid structure was crucial for the observed cells behavior. Although not completely understood, the encouraging results obtained constitute an incentive for further studies on this topic.

Baghdadite coating formed by hybrid water-stabilized plasma spray for bioceramic applications: Mechanical and biological evaluations

This work studies the mechanical and biological properties of Baghdadite (BAG, Ca3ZrSi2O9) coating manufactured on Ti6Al4V substrates by hybrid water-stabilized plasma spray (WSP-H). Hydroxyapatite (HAp, Ca10(PO4)6(OH)2) coating was produced by gas-stabilized atmospheric plasma spray and used as a reference material. Upon spraying, the BAG coating exhibited lower crystallinity than the HAp coating. Mechanical testing demonstrated superior properties of the BAG coating: its higher hardness, elastic modulus as well as a better resistance to scratch and wear. In the cell viability study, the BAG coating presented better human osteoblast attachment and proliferation on the coating surface after three days and seven days compared to the HAp counterpart. Furthermore, the gene expression study of human osteoblasts indicated that the BAG coating surface showed higher expression levels of osteogenic genes than those on the HAp coating. Overall, this study indicates that enhanced mechanical and bioactive properties can be achieved for the BAG coating compared to the benchmark HAp coating. It is therefore concluded here that the BAG coating is a potential candidate for coating orthopedic implants.

Selective Laser Melting and Electron Beam Melting of Ti6Al4V for Orthopedic Applications: A Comparative Study on the Applied Building Direction.

The 3D printing process offers several advantages to the medical industry by producing complex and bespoke devices that accurately reproduce customized patient geometries. Despite the recent developments that strongly enhanced the dominance of additive manufacturing (AM) techniques over conventional methods, processes need to be continually optimized and controlled to obtain implants that can fulfill all the requirements of the surgical procedure and the anatomical district of interest. The best outcomes of an implant derive from optimal compromise and balance between a good interaction with the surrounding tissue through cell attachment and reduced inflammatory response mainly caused by a weak interface with the native tissue or bacteria colonization of the implant surface. For these reasons, the chemical, morphological, and mechanical properties of a device need to be designed in order to assure the best performances considering the in vivo environment components. In particular, complex 3D geometries can be produced with high dimensional accuracy but inadequate surface properties due to the layer manufacturing process that always entails the use of post-processing techniques to improve the surface quality, increasing the lead times of the whole process despite the reduction of the supply chain. The goal of this work was to provide a comparison between Ti6Al4V samples fabricated by selective laser melting (SLM) and electron beam melting (EBM) with different building directions in relation to the building plate. The results highlighted the influence of the process technique on osteoblast attachment and mineralization compared with the building orientation that showed a limited effect in promoting a proper osseointegration over a long-term period.

UV-Pre-Treated and Protein-Adsorbed Titanium Implants Exhibit Enhanced Osteoconductivity.

Titanium materials are essential treatment modalities in the medical field and serve as a tissue engineering scaffold and coating material for medical devices. Thus, there is a significant demand to improve the bioactivity of titanium for therapeutic and experimental purposes. We showed that ultraviolet light (UV)-pre-treatment changed the protein-adsorption ability and subsequent osteoconductivity of titanium. Fibronectin (FN) adsorption on UV-treated titanium was 20% and 30% greater after 1-min and 1-h incubation, respectively, than that of control titanium. After 3-h incubation, FN adsorption on UV-treated titanium remained 30% higher than that on the control. Osteoblasts were cultured on titanium disks after 1-h FN adsorption with or without UV-pre-treatment and on titanium disks without FN adsorption. The number of attached osteoblasts during the early stage of culture was 80% greater on UV-treated and FN-adsorbed (UV/FN) titanium than on FN-adsorbed (FN) titanium; osteoblasts attachment on UV/FN titanium was 2.6- and 2.1-fold greater than that on control- and UV-treated titanium, respectively. The alkaline phosphatase activity of osteoblasts on UV/FN titanium was increased 1.8-, 1.8-, and 2.4-fold compared with that on FN-adsorbed, UV-treated, and control titanium, respectively. The UV/FN implants exhibited 25% and 150% greater in vivo biomechanical strength of bone integration than the FN- and control implants, respectively. Bone morphogenetic protein-2 (BMP-2) adsorption on UV-treated titanium was 4.5-fold greater than that on control titanium after 1-min incubation, resulting in a 4-fold increase in osteoblast attachment. Thus, UV-pre-treatment of titanium accelerated its protein adsorptivity and osteoconductivity, providing a novel strategy for enhancing its bioactivity.

Hydroxyapatite and β-TCP modified PMMA-TiO2 and PMMA-ZrO2 coatings for bioactive corrosion protection of Ti6Al4V implants.

Organic-inorganic hybrid coatings deposited on different types of metallic alloys have shown outstanding anticorrosive performance. The incorporation of osteoconductive additives such as hydroxyapatite (HA) and β-tricalcium phosphate (β-TCP) into organic-inorganic hybrid coatings is promising to improve the osseointegration and corrosion resistance of Ti6Al4V alloys, which are the most widely used metallic orthopedic and dental implant materials today. Therefore, this study evaluated the capability of poly(methyl methacrylate) (PMMA)-TiO2 and PMMA-ZrO2 hybrid coatings modified with HA and β-TCP to act as bioactive and corrosion protection coatings for Ti6Al4V alloys. In terms of cell growth and mineralization, osteoblast viability, Ca+2 deposition and alkaline phosphatase assays revealed a significant improvement for the HA and β-TCP modified coatings, compared to the bare alloy. This can be explained by an increase in nanoscale roughness and associated higher surface free energy, which lead to enhanced protein adsorption to promote osteoblast attachment and functions on the coatings. The effect of HA and β-TCP additives on the anticorrosive efficiency was studied by electrochemical impedance spectroscopy (EIS) in a simulated body fluid (SBF) solution. The coatings presented a low-frequency impedance modulus of up to 430GΩcm2, 5 decades higher than the bare Ti6Al4V alloy. These findings provide clear evidence of the beneficial role of HA and β-TCP modified hybrid coatings, improving both the biocompatibility and corrosion resistance of the Ti6Al4V alloy.

Fabrication of biocompatible and bioabsorbable polycaprolactone/ magnesium hydroxide 3D printed scaffolds: Degradation and in vitro osteoblasts interactions

Biodegradable polymeric 3D implants are of considerable interest for biomedical applications, however the degradation profile and bioactivity are important considerations for many clinical applications. In this context, bioresorbable magnesium hydroxide (MH) nanoparticles (NPs) (<50 nm) were blended with the degradable polymer poly (ε-caprolactone) (PCL) at concentrations of 5 and 20 wt%, and the composite was manufactured by 3D printing technology. Efficient load transfer was found between the nanofiller and matrix PCL, which was reflected in changes in the tensile properties of the MH-based composite. A statistically significant 44.3% increase in tensile modulus was achieved by the addition of 5 wt% MH, which was in agreement with the Halpin-Tsai theoretical model. The incorporation of MH in the PCL scaffolds accelerated the weight loss of the scaffolds and decreased the molecular weight of PCL over a prolonged soaking period (150 days) in PBS solution (pH 7.37, 37 ± 0.5 °C). The PCL/MH composite scaffolds were shown to be non-cytotoxic in vitro, and ion diffusion into the cell culture media promoted osteoblast metabolic activity, attachment, and proliferation, as compared to PCL-only scaffolds. Moreover, osteoblastic activity, as assessed by the expression of alkaline phosphatase, was significantly higher on the composite PCL/MH scaffold after 14 and 21 days. In summary, the 3D PCL/MH composite scaffolds could enhance osteoblastic activity and demonstrated a moderately accelerated degradation profile, which are characteristics that can be considered favorable for bone regeneration applications.

Engineering microstructure of hydroxyapatite by electron beam irradiation to induce controllable in vitro degradation

Controllable degradation behavior of hydroxyapatite (HA) plays a crucial role in successful long-term implantation of a bone tissue substitute. In this study, electron beam irradiation (EBI) was carried out as an efficient strategy to modify HA structure. The degradation characteristics, bioactivity and cell response of the irradiated HA were comprehensively evaluated. The results showed that EBI with absorbed dose of 3.42, 10.27 and 20.54 Gy did not introduce newly-formed phase or impurity, but created crystal defects and nanovoids, which were formed from oxygen deficiencies, into HA lattice with irradiation dose-dependence. The dissolution rate of the irradiated HA enhanced continuously with increasing irradiation dose, as a result of constantly increasing crystal defects and enlarging voids, where the main reactive sites for the degradation of HA formed. The Ca2+ and PO43- releasing of HA with the highest irradiation dose enhanced by 92.75% and 138.85% respectively compared to that of the pristine HA after 28 days’ immersion. SBF immersion results suggested the irradiated HA with raising EBI doses exhibited preferable performance in bioactivity as well. Moreover, the results from CCK-8 assay and SEM observation showed that the irradiated HA were beneficial to the proliferation and attachment of osteoblasts.

Osteoblasts migration, attachment and human bone marrow-mesenchymal stem cells osteogenic differentiation towards surface engineered and growth factors conjugated poly(lactic acid) microspheres.

Recently, surface engineered biomaterials through surface modification are extensively investigated due to its potential to enhance cellular homing and migration which contributes to a successful drug delivery process. This study is focused on osteoblasts response towards surface engineered using a simple sodium hydroxide (NaOH) hydrolysis and growth factors conjugated poly(lactic acid) (PLA) microspheres. In this study, evaluation of the relationship of NaOH concentration with the molecular weight changes and surface morphology of PLA microspheres specifically wall thickness and porosity prior to in vitro studies was investigated. NaOH hydrolysis of 0.1 M, 0.3 M and 0.5 M were done to introduce hydrophilicity on the PLA prior to conjugation with basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF). Morphology changes showed that higher concentration of NaOH could accelerate the hydrolysis process as the highest wall thickness was observed at 0.5 M NaOH with ~3.52 µm. All surface modified and growth factors conjugated PLA microspheres wells enhanced the migration of the cells during wound healing process as wound closure was 100% after 3 days of treatment. Increase in hydrophilicity of the surface engineered and growth factors conjugated PLA microspheres provides favorable surface for cellular attachment of osteoblast, which was reflected by positive DAPI staining of the cells' nucleus. Surface modified and growth factors conjugated PLA microspheres were also able to enhance the capability of the PLA in facilitating the differentiation process of mesenchymal stem cells (MSCs) into osteogenic lineage since only positive stain was observed on surface engineered and growth factors conjugated PLA microspheres. These results indicated that the surface engineered and growth factors conjugated PLA microspheres were non-toxic for biological environments and the improved hydrophilicity made them a potential candidate as a drug delivery vehicle as the cells can adhere, attach and proliferate inside it.

Microstructures and cell reaction of porous hydroxyapatite coatings on titanium discs using a novel vapour-induced pore-forming atmospheric plasma spraying

The surface topography of implants plays an important role in affecting the behaviour of osteoblasts. Previous research has demonstrated that porous hydroxyapatite (HAp) coating surfaces fabricated by vapour-assisted flame spraying enhances osteoblast attachment and differentiation, but flame spraying cannot supply sufficient power for fabricating porous surfaces without cracks. To solve this problem, porous HAp coatings were deposited on titanium (Ti) discs using a novel vapour-induced pore-forming atmospheric plasma spraying (VIPF-APS) technique with a higher amount of energy. A water reservoir was placed behind the Ti discs to serve as the vapour source. Ti discs with three different mean pore diameters, namely, ATi (2.65 μm), BTi (15.65 μm), and CTi (35.33 μm), were used, and the HAp coatings that were deposited on these three discs using the VIPF-APS technique were denoted HAp-ATi, HAp-BTi, and HAp-CTi, respectively. For comparison, the conventional APS technique was used to deposit HAp on ATi, and this sample was named HAp-APS/ATi. The results showed that the VIPF-APS technique effectively produced porous HAp coatings, whereas the conventional APS technique yielded dense coatings that are unfavourable for applications. These porous coatings further contributed to higher osteoblast proliferation and distinctive alkaline phosphatase activity. The VIPF-APS technique thus has great potential for producing bioactive coatings that can be used in implants.

Impact of Laser Structuring on Medical-Grade Titanium: Surface Characterization and In Vitro Evaluation of Osteoblast Attachment.

Improved implant osteointegration offers meaningful potential for orthopedic, spinal, and dental implants. In this study, a laser treatment was used for the structuring of a titanium alloy (Ti6Al4V) surface combined with a titanium dioxide coating, whereby a porous surface was created. The objective was to characterize the pore structure shape, treatment-related metallographic changes, cytocompatibility, and attachment of osteoblast-like cells (MG-63). The treatment generated specific bottleneck pore shapes, offering the potential for the interlocking of osteoblasts within undercuts in the implant surface. The pore dimensions were a bottleneck diameter of 27 µm (SD: 4 µm), an inner pore width of 78 µm (SD: 6 µm), and a pore depth of 129 µm (SD: 8 µm). The introduced energy of the laser changed the metallic structure of the alloy within the heat-affected region (approximately 66 µm) without any indication of a micro cracking formation. The phase of the alloy (microcrystalline alpha + beta) was changed to a martensite alpha phase in the surface region and an alpha + beta phase in the transition region between the pores. The MG-63 cells adhered to the structured titanium surface within 30 min and grew with numerous filopodia over and into the pores over the following days. Cell viability was improved on the structured surface compared to pure titanium, indicating good cytocompatibility. In particular, the demonstrated affinity of MG-63 cells to grow into the pores offers the potential to provide significantly improved implant fixation in further in vivo studies.

In vitro evaluation of bilayer membranes of PLGA/hydroxyapatite/β-tricalcium phosphate for guided bone regeneration.

Membranes for guided bone regeneration represent valuable resources, preventing fibroblast infiltration and aiding anatomical bone reconstruction. Nonetheless, available membranes lack bone regenerative capacity, suitable mechanical behavior, or adequate degradation profile. Therefore, to overcome these limitations, this study developed bilayer membranes with a dense layer (dry phase inversion) of PLGA (poly(lactic-co-glycolic acid)):HAp (hydroxyapatite) - 95:05 (wt%) - and an electrospun layer of PLGA and HAp:β-TCP (β-tricalcium phosphate) with ratios of 60:40, 70:30 and 85:15 (wt%), evaluating its mechanical, morphological and in vitro properties. The bilayer membranes displayed adequate interlayer adhesion, dense layer pore size of 4.20μm and electrospun layer with porosity degree of 38.2%, thus capable of preventing fibroblast infiltration while allowing osteoblast migration and nutrient permeation. They also showed Tg of 82°C and higher storage modulus, which was constant up to 54.6°C, characteristics important for membrane implantation and use with no mechanical compromise. In vitro degradation mass loss was only 10% after 60days, a profile suitable for the application requirement. Membranes with calcium phosphates had better osteoblast attachment, proliferation and migration. Taken together, results indicate the great potential of PLGA/HAp/β-TCP bilayer membranes on bone reconstruction with proper degradation profile, morphology, mechanical behavior and bone regenerative capacity.

Bioactive Effects of Low-Temperature Argon-Oxygen Plasma on a Titanium Implant Surface.

Although titanium is the most commonly used dental implant material, its biological aging directly leads to a lower rate of osseointegration. The aim of this study is to treat aged titanium disc surfaces using low-temperature argon–oxygen plasma (LTAOP) to obtain a more hydrophilic surface in order to enhance biological activities of osteoblasts on dental implant materials. In this study, smooth-machined titanium (SM Ti) and sandblasted and acid-etched titanium (SLA Ti) substrates were used. Aged titanium discs (SM and SLA Ti) were activated by LTAOP and the surface properties were analyzed. Osteoblasts were then seeded onto the aged and LTAOP-treated surfaces. Cell morphology, viability, and features of osteogenesis were examined. We showed that after the LTAOP treatment, the surfaces of both SM and SLA titanium substrates become more hydrophilic with a larger active oxygen species composition, whereas no obvious morphological changes were observed. Osteoblasts were found to be attached and stretched well on the surfaces of LTAOP treatment specimens. Moreover, the proliferation and osteocalcin secretion of osteoblasts on the plasma-activated titanium samples were superior to the untreated counterparts. LTAOP activation can enhance the attachment, proliferation, and mineralization of osteoblasts on the surfaces of the aged titanium substrates. This research provides a new strategy to modify the surface of titanium dental implants for improved biological functions.

A laminin-211-derived bioactive peptide promotes the osseointegration of a sandblasted, large-grit, acid-etched titanium implant.

Early implant loading is very important for reducing the duration of missing teeth in human patients. The laminin-derived peptide, DLTIDDSYWYRI motif (Ln2-P3), accelerates bone healing. Therefore, to investigate the hypothesis that Ln2-P3 increases the bone response to sandblasted, large-grit, acid-etched (SLA) titanium implants, the effect of the Ln2-P3 peptide on the osseointegration of SLA titanium implants was evaluated in vitro and in vivo. Human osteoblast-like cells were cultured on untreated, scrambled peptide (SP)-treated, and Ln2-P3-treated SLA titanium discs, and the cellular responses of these cells were evaluated. The Ln2-P3 treatment augmented osteoblast attachment and spreading, alkaline phosphatase activity, and the expression of osteogenic marker genes. Furthermore, the untreated and Ln2-P3-treated SLA titanium implants were inserted into the tibiae of rabbits for 9 and 11 days. Compared with the untreated implants, the Ln2-P3-treated implants showed a significantly higher bone-to-implant contact ratio at Day 9 after implantation and an increased bone area. The Ln2-P3 treatment of the SLA titanium implant surface augmented osteoblastic activity and accelerated peri-implant bone formation at the bone-implant interface. Overall, these results indicated that compared with the SLA titanium surface alone, the Ln2-P3 peptide-treated SLA titanium surface enhances initial osseointegration, thereby facilitating earlier implant loading.

Surface mechanical attrition treatment of low modulus Ti-Nb-Ta-O alloy for orthopedic applications.

Surface mechanical attrition treatment (SMAT) is recognized as a surface severe plastic deformation (SPD) method that is effective in improving the surface-dependent mechanical and functional properties of conventional metallic biomaterials. In this study, we aimed to systemically investigate the effect of SMAT on the physical, electrochemical, tribological and biological performances of a newly developed low modulus β Ti-Nb-Ta-O alloy with two different microstructures, namely, single phase β-treated and dual phase β+α aged. The microhardness results showed considerable hardening for the β-treated condition due to formation of deformation substructures; that was associated with increased corrosion resistance resulting from a stronger and denser passive layer on the surface, as revealed by Tafel polarization, impedance studies and Mott-Scottky plots. The wear volume loss during fretting in serum solution was found to decrease by 46% while friction coefficient decreased only marginally, due to presence of a harder and more brittle surface. In the β+α condition of the alloy, minimal hardening was observed due to coarsening of the precipitates during SMAT. However, this also reduced the number of α-β interfaces, which in turn minimized the tendency for galvanic corrosion resulting in lower corrosion rate after SMAT. Wear resistance was enhanced after SMAT, with 32% decrease in wear volume loss and 21% decrease in friction coefficient resulted due to improved ductility on the surface. The attachment and growth of osteoblasts on the alloys in vitro were not affected by SMAT and was comparable to that on commercially pure Ti. Taken together, these results provide new insights into the effects of surface SPD of low modulus β- Ti alloys for orthopedic applications and underscore the importance of the initial microstructure in determining the performance of the alloy.

A Newly Created Meso-, Micro-, and Nano-Scale Rough Titanium Surface Promotes Bone-Implant Integration.

Titanium implants are the standard therapeutic option when restoring missing teeth and reconstructing fractured and/or diseased bone. However, in the 30 years since the advent of micro-rough surfaces, titanium’s ability to integrate with bone has not improved significantly. We developed a method to create a unique titanium surface with distinct roughness features at meso-, micro-, and nano-scales. We sought to determine the biological ability of the surface and optimize it for better osseointegration. Commercially pure titanium was acid-etched with sulfuric acid at different temperatures (120, 130, 140, and 150 °C). Although only the typical micro-scale compartmental structure was formed during acid-etching at 120 and 130 °C, meso-scale spikes (20–50 μm wide) and nano-scale polymorphic structures as well as micro-scale compartmental structures formed exclusively at 140 and 150 °C. The average surface roughness (Ra) of the three-scale rough surface was 6–12 times greater than that with micro-roughness only, and did not compromise the initial attachment and spreading of osteoblasts despite its considerably increased surface roughness. The new surface promoted osteoblast differentiation and in vivo osseointegration significantly; regression analysis between osteoconductivity and surface variables revealed these effects were highly correlated with the size and density of meso-scale spikes. The overall strength of osseointegration was the greatest when the acid-etching was performed at 140 °C. Thus, we demonstrated that our meso-, micro-, and nano-scale rough titanium surface generates substantially increased osteoconductive and osseointegrative ability over the well-established micro-rough titanium surface. This novel surface is expected to be utilized in dental and various types of orthopedic surgical implants, as well as titanium-based bone engineering scaffolds.

Tuning of Titanium Microfiber Scaffold with UV-Photofunctionalization for Enhanced Osteoblast Affinity and Function.

Titanium (Ti) is an osteoconductive material that is routinely used as a bulk implant to fix and restore bones and teeth. This study explored the effective use of Ti as a bone engineering scaffold. Challenges to overcome were: (1) difficult liquid/cell infiltration into Ti microfiber scaffolds due to the hydrophobic nature of Ti; and (2) difficult cell attachment on thin and curved Ti microfibers. A recent discovery of UV-photofunctionalization of Ti prompted us to examine its effect on Ti microfiber scaffolds. Scaffolds in disk form were made by weaving grade 4 pure Ti microfibers (125 µm diameter) and half of them were acid-etched to roughen the surface. Some of the scaffolds with original or acid-etched surfaces were further treated by UV light before cell culture. Ti microfiber scaffolds, regardless of the surface type, were hydrophobic and did not allow glycerol/water liquid to infiltrate, whereas, after UV treatment, the scaffolds became hydrophilic and immediately absorbed the liquid. Osteogenic cells from two different origins, derived from the femoral and mandibular bone marrow of rats, were cultured on the scaffolds. The number of cells attached to scaffolds during the early stage of culture within 24 h was 3–10 times greater when the scaffolds were treated with UV. The development of cytoplasmic projections and cytoskeletal, as well as the expression of focal adhesion protein, were exclusively observed on UV-treated scaffolds. Osteoblastic functional phenotypes, such as alkaline phosphatase activity and calcium mineralization, were 2–15 times greater on UV-treated scaffolds, with more pronounced enhancement on acid-etched scaffolds compared to that on the original scaffolds. These effects of UV treatment were associated with a significant reduction in atomic carbon on the Ti microfiber surfaces. In conclusion, UV treatment of Ti microfiber scaffolds tunes their physicochemical properties and effectively enhances the attachment and function of osteoblasts, proposing a new strategy for bone engineering.

A nitrogen-containing bisphosphonate inhibits osteoblast attachment and impairs bone healing in bone-compatible scaffold

Compromised osteoblast attachment on hydroxyapatite could be involved in the development of bone healing failure. We developed a bone-compatible scaffold that mimics bone structure with sub-micron hydroxyapatite (HA) surfaces, so that we could evaluate the effects of nitrogen-containing bisphosphonate (N-BP) on osteoblast behavior and bone healing. Human osteoblasts were seeded onto the bone-compatible scaffold with or without N-BP, and cell attachment and spreading behavior were evaluated 4 and 24 h after seeding. Then, mineralization was evaluated at 7 and 14 days. The osteoconductive activity of the scaffold was evaluated by implantation for 3 and 6 weeks into a rat cranial bone defect. The numbers of osteoblasts and their diameters were significantly less in N-BP-binding scaffolds than in untreated scaffolds at 4 and 24 h. Mineralization were also significantly less in the N-BP-binding scaffolds than in controls at 7 and 14 days. In vivo study revealed bone formation in N-BP-binding scaffolds was significantly less than in untreated scaffolds at 3 and 6 weeks. These results suggest that N-BP-binding to HA inhibited osteoblast attachment and spreading, thereby compromising bone healing process in the injured bone defect site.

A novel CPC composite cement reinforced by dopamine coated SCPP fibers with improved physicochemical and biological properties.

Traditional CPC cements have attracted wide attentions in repairing bone defects for injectability, easy plasticity and good osseointegration. However, its further application was limited by poor mechanical properties, long setting time and unsatisfactory biocompatibility. To solve these problems, polydopamine (DOPA) coated strontium-doped calcium polyphosphate (SCPP) fibers were added into CPC cements for the first time. A doping amount at fiber weight fraction of 0%, 1%, 2% and 5% was designed to develop a multifunctional composite fitting for bone tissues' regeneration and reconstruction and the optimum amount was selected through subsequent physicochemical and biological characterizations. The results implied DOPA coating successfully formed stable connections between SCPP fibers and CPC matrix, which simultaneously reinforced biomechanical strength and tenacity (5% SCPP/D/CPC samples exhibited more prominent mechanical property than others). In addition, 5% D/SCPP fibers doped composite cements were characterized as markedly-improved cytocompatibility: Sr2+ introduction induced cytoactive and significantly accelerated proliferation, attachment and spreading of osteoblasts. Besides, it also stimulated the secretion of OT, Col-I and ALP from seeded MG63, which was a critical character for further inducing osteogenic process, mineralization and bone tissues formation. The promoted cytocompatibility and improved osteogenesis-related growth factors' secretion could be attributed to constant and controllable release of Sr2+ and this deduction was approved by ICP analysis. In addition, Sr doping made this novel cement had a potential efficacy to inhibit aseptic loosening. In a word, present studies all demonstrated 5% SCPP/D/CPC composites could be a potential candidate material employed in bone regeneration and reconstruction for excellent mechanical property and cytocompatibility.