Quercetin modified electrospun PHBV fibrous scaffold enhances cartilage regeneration.

It suggests that the poly (3-hydroxybutyric acid-co-3-hydroxyvaleric acid) (PHBV) scaffold can be used for cartilage tissue engineering, but PHBV is short of bioactivity that is required for cartilage regeneration. To fabricate a bioactive cartilage tissue engineering scaffold that promotes cartilage regeneration, quercetin (QUE) modified PHBV (PHBV-g-QUE) fibrous scaffolds were prepared by a two-step surface modification method. The PHBV-g-QUE fibrous scaffold facilitates the growth of chondrocytes and maintains chondrocytic phenotype resulting from the upregulation of SOX9, COL II, and ACAN. The PHBV-g-QUE fibrous scaffold inhibited apoptosis of chondrocyte and reduced oxidative stress of chondrocytes by regulating the transcription of related genes. Following PHBV-g-QUE fibrous scaffolds and PHBV fibrous scaffolds with adhered chondrocytes were implanted into nude mice for 4 weeks, it demonstrated that PHBV-g-QUE fibrous scaffolds significantly promoted cartilage regeneration compared with the PHBV fibrous scaffolds. Hence, it suggests that the PHBV-g-QUE fibrous scaffold can be potentially applied in the clinical treatment of cartilage defects in the future.

Decellularized sturgeon cartilage extracellular matrix scaffold inhibits chondrocyte hypertrophy in vitro and in vivo.

Since chondrocyte hypertrophy greatly limits the efficiency of cartilage defects repairing via cartilage tissue engineering (CTE), it is critical to develop a functional CTE scaffold able to inhibit chondrocyte hypertrophy during this period of cartilage regeneration. In this study, we tested the applicability of using decellularized sturgeon cartilage ECM (dSCECM) scaffold to cease chondrocyte hypertrophy during cartilage damage repair. The dSCECM scaffolds with interconnected porous structure and pore size of 114.1 ± 20.9 µm were successfully prepared with freeze-dry method. Chondrocytes displayed a round shape and aggregated to form cellular spheroids within dSCECM scaffolds, which is similar to their chondrocytic phenotype within cartilage in vivo. Higher transcriptional level of chondrogenic related genes and integrin related genes was observed in chondrocytes incubated with dSCECM scaffolds instead of type I collagen (COL I) scaffolds, which were used as the control due to their widely usage in CTE and clinic applications. Furthermore, it confirmed that, compared with COL I scaffolds, dSCECM scaffolds significantly reduced the transcription of chondrocyte hypertrophy related genes in chondrocytes following the hypertrophic induction treatment. To test the ability of dSCECM scaffold to inhibit chondrocytes hypertrophy in vivo, chondrocytes with dSCECM scaffolds and COL I scaffolds were cultured with hypertrophic media and were implanted into nude mice respectively. Following 4 weeks implantation, interestingly, only the specimens derived from COL I scaffolds displayed consequences of chondrocyte hypertrophy like calcification deposition, demonstrating that chondrocyte hypertrophy is ceased by the dSCECM scaffold following hypertrophic induction. It suggests that the dSCECM scaffold can be potentially applied in clinical treating cartilage defects via the CTE approach to avoid the risk of chondrocyte hypertrophy.

Effect of extrusion process on the mechanical and in vitro degradation performance of a biomedical Mg-Zn-Y-Nd alloy.

A new type of biomedical Mg-Zn-Y-Nd alloy was developed and thermal extruded by different processes to investigate the effect of extrusion ratio and extrusion pass on its microstructure, mechanical property and degradation performance. The results show that the increase of extrusion ratio could promote the dynamic recrystallization (DRX) process and led to the coarsening of DRXed grains. While the increase of extrusion pass also contributes to the DRX process but refines the DRXed grains. The simultaneous increasing of extrusion ratio and extrusion pass refines the secondary phases obviously. The increase of extrusion ratio has reduced the tensile strength but improved the elongation of the alloy significantly. However, the increase of extrusion pass could enhance the tensile strength and elongation simultaneously, especially the strength. The degradation performance has been optimized effectively through increasing the extrusion ratio and extrusion pass.

A tannic acid-modified fluoride pre-treated Mg-Zn-Y-Nd alloy with antioxidant and platelet-repellent functionalities for vascular stent application.

Vascular stent interventional therapy, as a regular and effective therapy, has been widely used to treat coronary artery diseases. However, adverse events occur frequently after stent intervention, especially restenosis and late stent thrombosis. The targeted implanting site will suffer from severe atherosclerosis, which is considered as a chronic inflammatory disease. Meanwhile, with the over-expanding use of endovascular mechanical intervention, vascular injury has become an increasingly common issue. Lesions and newly induced vascular injury result in inflammatory and oxidative stress; meanwhile, activated macrophages and granulocytes generate high levels of reactive oxygen species (ROS), contributing to endothelial dysfunction and neointima hyperplasia. Therefore, attenuating oxidative stress and reducing ROS generation in the inflammatory response represent reasonable strategies to inhibit intimal hyperplasia and restenosis. Herein, we have developed a multifunctional surface for the MgZnYNd alloy with tannic acid (TA) coating, and the pH dependence of the coating deposition is also demonstrated. The phenolic hydroxyl groups on the coatings endow the modified surface with excellent antioxidant functions. We found that the coating can be recycled, and the scavenging activity hardly weakened within five cycles. Also, the TA coating has a promising strong antioxidant activity as it shows a radical scavenging activity over 80% in long term. Moreover, the TA coating possesses platelet-repellent capability. No significant inflammatory response was observed for the TA modified sample in the rat subcutaneous implantation test. Combining these performances, we envision that the vascular stent modified with TA coating can have great potential in various applications by virtue of its simplicity and effectiveness.

In Vitro and in Vivo Studies on Two-Step Alkali-Fluoride-Treated Mg-Zn-Y-Nd Alloy for Vascular Stent Application: Enhancement in Corrosion Resistance and Biocompatibility.

Bioabsorbable magnesium alloys are becoming prominent materials for cardiovascular stents, as their desirable mechanical properties and favorable biosafety. However, the rapid corrosion of magnesium alloys under physiological conditions hinders their wider application as medical implant materials. Fluoride chemical conversion treatment is an effective and simple technique to improve the corrosion resistance for magnesium alloys. Despite previous literature reporting on fluoride chemical conversion treatment with hydrofluoric acid (HF) in different conditions, some defects are still present on the surface of the coating. In this study, we report on a two-step alkali-fluoride treatment of magnesium alloy by effectively removing the second phase in the substrate surface and form a dense and flawless magnesium fluoride (MgF2) coating to endow the magnesium alloy greater corrosion resistance. The results showed that the serious pitting corrosion caused by galvanic corrosion could be effectively prevented after removing of the second phase of the surface. In vivo tests in a rat subcutaneous implantation model showed that two-step alkali-fluoride-treated MgZnYNd alloy (MgZnYNd-A-F) uniformly corroded with a low corrosion rate. No subcutaneous gas cavities or significant inflammatory cell infiltration were observed for MgZnYNd-A-F in in vivo tests. The two-step alkali-fluoride treatment can significantly improve the corrosion resistance and biocompatibility of magnesium alloy, which has great potential in the application of vascular stents because of its simplicity and effectiveness.

An electrospun fiber reinforced scaffold promotes total meniscus regeneration in rabbit meniscectomy model.

Low vascularization in meniscus limits its regeneration ability after injury, and tissue engineering is the most promising method to achieve meniscus regeneration. In this study, we fabricated a kind of composite scaffold by decellularized meniscus extracellular matrix/polycaprolactone (DMECM/PCL) electrospinning fibers and porous DMECM, in which DMECM/PCL fibers were used as reinforcing component. The tensile modulus of the composite scaffold in longitudinal and crosswise directions were 8.5 ±â€¯1.9 and 2.3 ±â€¯0.3 MPa, respectively. Besides that, the DMECM/PCL electrospinning fibers enhanced suture resistance of the composite scaffold more than 5 times than DMECM scaffold effectively. In vitro cytocompatibility showed that the porous structure provided by DMECM component facilitated meniscus cells' proliferation. DMECM was also the main component to regulate cell behaviors, which promoted meniscus cells expressing extracellular matrix related genes such as COL I, COL II, SOX9 and AGG. Rabbits with total meniscectomy were used as animal model to evaluated the composited scaffolds performance in vivo at 3 and 6 months. Results showed that rabbits with scaffold implanting could regenerate neo-menisci in both time points. The neo-menisci had similar histology structure and biochemical content with native menisci. Although neo-menisci had inferior tensile modulus than native ones, its modulus was improved with implanting time prolonging. MRI imaging showed the signal of neo-meniscus in the body is clear, and X-ray imaging of knee joints demonstrated the implantation of scaffolds could relief joint space narrowing. Moreover, rabbits with neo-menisci had better cartilage condition in femoral condyle and tibial plateau compared than meniscectomy group. STATEMENT OF SIGNIFICANCE: We fabricated the meniscus scaffold by combining porous decellularized meniscus extracellular matrix (DMECM) and DMECM/PCL electrospinning fibers together, which used the porous structure of DMECM, and the good tensile property of electrospinning fibers. We believe single material cannot satisfy increasing needs of scaffold. Therefore, we combined not only materials but also fabrication methods together to develop scaffold to make good use of each part. DMECM in electrospinning fibers also made these two components possible to be integrated through crosslinking. Compared to existing meniscus scaffold, the composite scaffold had (1) soft structure and extrusion would not happen after implantation, (2) ability to be trimmed to suitable shape during surgery, and (3) good resistance to suture.

A high-throughput study on endothelial cell adhesion and growth mediated by adsorbed serum protein via signaling pathway PCR array.

The purpose of this paper is to utilize the signaling pathway polymerase chain reaction (PCR) arrays to investigate the activation of two important biological signaling pathways in endothelial cell adhesion and growth mediated by adsorbed serum protein on the surface of bare and titanium nitride (TiN)-coated nickel titanium (NiTi) alloys. First, the endothelial cells were cultured on the bare and TiN-coated NiTi alloys and chitosan films as control for 4 h and 24 h, respectively. Then, the total RNA of the cells was collected and the PCR arrays were performed. After that, the differentially expressed genes in the transforming growth factor beta (TGF-ß) signaling pathway and the regulation of actin cytoskeleton pathway were screened out; and the further bioinformatics analyses were performed. The results showed that both TGF-ß signaling pathway and regulation of actin cytoskeleton pathway were activated in the cells after 4 h and 24 h culturing on the surface of bare and TiN-coated NiTi alloys compared to the chitosan group. The activated TGF-ß signaling pathway promoted cell adhesion; the activated regulation of actin cytoskeleton pathway promoted cell adhesion, spreading, growth and motility. In addition, the activation of both pathways was much stronger in the cells cultured for 24 h versus 4 h, which indicated that cell adhesion and growth became more favorable with longer time on the surface of two NiTi alloy materials.

The application of electrospinning used in meniscus tissue engineering.

Meniscus is a fibrocartilaginous organ to redistribute stress and enhance the stability of knee joint. Meniscus injury is common and still a formidable challenge to orthopedic surgeons. Surgical techniques and allograft transplantation were primary approaches to meniscus repair, but with intrinsic limitations in clinical practice. Tissue engineering is the most promising method to repair meniscus at present. Electrospinning is a method to fabricate fibers in small scale. With different materials and parameters, electrospinning materials could have different mechanical properties, porosity, and orientation, which could mimic architectural features and mechanical properties of native meniscus. Therefore, electrospinning materials could be used in meniscus regeneration and curing. This review gave a brief introduction of meniscus structure, injury, treatment and the application of electrospinning fibers in meniscus tissue engineering and curing. Besides that, we summarized materials commonly used in electrospinning to fabricate meniscus scaffolds, and discussed the form of electrospinning fibers used such as scaffold, substitute and patch. Finally, the function of electrospinning fibers, for example, carrying drugs, providing mechanical properties were described. The potential applications of electrospinning fibers in meniscus therapy were proposed.

A silk-based coating containing GREDVY peptide and heparin on Mg-Zn-Y-Nd alloy: improved corrosion resistance, hemocompatibility and endothelialization.

Magnesium (Mg) alloys have been intensively investigated as potential absorbable coronary stent materials as their use avoids risks such as late inflammation and restenosis generated by permanent metallic implants. Besides that, clinical trials on coronary stents fabricated from Mg alloys have made great progress recently. However, the over-rapid corrosion rate, magnesium corrosion-induced thrombosis formation and delayed endothelium regeneration continue to be problematic for coronary artery stent therapy. In this study, silk fibroin blended with heparin and GREDVY (Gly-Arg-Glu-Asp-Val-Tyr) peptide was immobilized on a HF-pretreated MgZnYNd alloy surface via a polydopamine layer to improve its corrosion resistance, blood compatibility and endothelialization. Standard electrochemical measurements along with the long-term immersion results indicated that the functionalized MgZnYNd alloy had preferable anti-corrosion abilities compared with the bare MgZnYNd alloy. The modified surface exhibited outstanding hemocompatibility with reduced platelet adhesion, hemolysis rate and prolonged blood coagulation time. Human umbilical vein endothelial cell (HUVEC) and vascular smooth muscle cell (VSMC) co-culture results revealed more attached HUVECs on the functionalized samples than on the MgZnYNd alloy surfaces. The excellent corrosion retardation, hemocompatibility and re-endothelialization of the multi-functional coating indicate a promising method in the field of biodegradable magnesium-based implantable cardiovascular stents.

Arginine-leucine based poly (ester urea urethane) coating for Mg-Zn-Y-Nd alloy in cardiovascular stent applications.

Selected from the family of self-designed biodegradable amino acid-based poly (ester urea urethane) (AA-PEUU) pseudo-protein biomaterials, arginine-leucine based poly (ester urea urethane)s (Arg-Leu-PEUUs) were used as protective and bio-functional coatings for bio-absorbable magnesium alloy MgZnYNd in cardiovascular stent applications. Comparing with poly (glycolide-co-lactide) (PLGA) coating, the Arg-Leu-PEUU coating had stronger bonding strength with the substrate; in vitro electrochemical and long-term immersion results verified a significantly better corrosion resistance. Acute blood contact tests proved a better hemocompatibility of Arg-Leu-PEUU coating. The immunofluorescent staining and cell proliferation test indicated that Arg-Leu-PEUU coating had a far better cytocompatibility. The Arg-Leu-PEUU coating stimulated human umbilical vein endothelial cells (HUVEC) to release reasonably increased amount of nitric oxide (NO), suggesting its potential in retarding thrombosis and restenosis. The superior corrosion resistance and biocompatibility as well as the indigenous NO production bio-functionality of the Arg-Leu-PEUU copolymer family indicate their capability to offer a far better protection of the magnesium-based implantable cardiovascular stent and bring their application closer to clinical reality.

Functionalized Polymeric Membrane with Enhanced Mechanical and Biological Properties to Control the Degradation of Magnesium Alloy.

To achieve enhanced biological response and controlled degradation of magnesium alloy, a modified biodegradable polymer coating called polycaprolactone (PCL) is fabricated by a thermal approach in which the heat treatment neither alters the chemical composition of the PCL membrane nor the rate of magnesium ion release, pH value, or weight loss, compared with the untreated sample. The changes in the crystallinity, hydrophilicity, and oxygen content of heat-treated PCL coating not only improve the mechanical adhesion strength between the coating and magnesium substrate but also enhance the biological properties. Moreover, the thermally modified sample can lead to higher spreading and elongation of osteoblasts, due to the enhanced hydrophilicity and CO to CO functional group ratio. In the analyses of microcomputed tomography from one to four weeks postoperation, the total volume of new bone formation on the heat-treated sample is 10%-35% and 70%-90% higher than that of the untreated and uncoated controls, respectively. Surprisingly, the indentation modulus of the newly formed bone adjacent to the heat-treated sample is ≈20% higher than that of both controls. These promising results reveal the clinical potential of the modified PCL coating on magnesium alloy in orthopedic applications.

Fabrication and characterization of electrospun nanofibers composed of decellularized meniscus extracellular matrix and polycaprolactone for meniscus tissue engineering.

Many kinds of scaffolds have been produced in meniscus tissue engineering, but few have matched the mechanical properties of native meniscus, making it impossible for them to sustain large stress at initial implantation. In this study, we used a differential centrifugation method to obtain decellularized meniscus extracellular matrix (DMECM) and combined the DMECM with polycaprolactone (PCL) via electrospinning to fabricate random and aligned microfibers. The FTIR results and biochemical assays demonstrated the successful mixing of these two elements, and the addition of DMECM improved the hydrophilicity of the microfibers. The blending of DMECM also enhanced the tensile modulus of the microfibers, and aligned fibers had tensile moduli ranging from 132.27 to 331.40 MPa, which match that of human meniscus. In addition, we defined yield stress as the lose-efficacy point. The results showed that DMECM/PCL fibers had higher yield stresses than the pure PCL fibers, and the aligned fibers had higher yield stress values than the randomly oriented fibers. Nanoindentation results showed that adding DMECM had no significant impact on modulus and hardness with the exception of fibers containing 80% DMECM, which exhibited an obvious increase in modulus. In vitro assay demonstrated that the DMECM/PCL fibers had no hemolysis or cytotoxicity. Meniscus cells could attach and proliferate on the fibers, and the fiber orientation had a direct influence on cell arrangement. RT-PCR results showed that meniscus cells had higher gene expressions of aggrecan, collagen I, collagen II and Sox 9 when seeded on fibers with higher DMECM contents.

A novel biodegradable and biologically functional arginine-based poly(ester urea urethane) coating for Mg-Zn-Y-Nd alloy: enhancement in corrosion resistance and biocompatibility.

A novel family of biodegradable pseudo-protein biomaterials, arginine (Arg)-based poly(ester urea urethane) (Arg-PEUU), were synthesized and applied as a better protective and bio-functional coating for bio-absorbable magnesium alloy MgZnYNd as a stent model. The Arg-PEUU coatings were stronger than poly(glycolide-co-lactide) (PLGA) coating with 11.9-103.4% higher critical lateral force. Electrochemical tests and in vitro immersion results demonstrated that the Arg-PEUU-coated MgZnYNd alloys have a significantly better corrosion resistance. The Arg-PEUU coating also showed reduced platelet adhesion and hemolysis rate in acute blood contact testing. Immunofluorescent actin and vinculin stainings showed that the Arg-PEUU coating had a far better cell adhesion of human umbilical vein endothelial cells (HUVEC), and also showed no cytotoxicity toward both HUVEC and human aortic smooth muscle cells (HASMC). The Arg-PEUU coating stimulated HUVEC to release significantly higher amounts of nitric oxide (NO) than the controls, suggesting the Arg-PEUU coating has the ability to retard thrombus and restenosis. The superb corrosion retardation, hemocompatibility and cytocompatibility of the Arg-PEUU coating as well as its induced indigenous NO production biofunctionality indicate that the newly developed Arg-PEUU biodegradable copolymer family may have the potential to offer a far greater protection of magnesium-based implantable cardiovascular stents.

Comparison of glutaraldehyde and carbodiimides to crosslink tissue engineering scaffolds fabricated by decellularized porcine menisci.

The objectives of this study were to fabricate porous scaffolds using decellularized meniscus, and to explore a preferable crosslinking condition to enhance mechanical properties of scaffolds. Moreover, the microstructure, porosity, biodegradation and cytotoxicity were also evaluated. EDAC or GTA in different concentration was used to crosslink scaffolds. FTIR demonstrated functional groups change in crosslinking process. SEM photography showed that crosslinked scaffolds had blurry edges, which resulted scaffolds crosslinked by 1.2mol/l EDAC had smaller porosity than other groups. The structure change enhanced antidegradation property. After immersing in enzyme solution for 96h, scaffolds crosslinked by GTA and EDAC could maintain their mass >70% and 80%. Most importantly, mechanical properties of crosslinked scaffolds were also improved. Uncrosslinked Scaffolds had only 0.49kPa in compression modulus and 12.81kPa in tensile modulus. The compression and tensile modulus of scaffolds crosslinked by 1.0% GTA were 1.42 and 567.44kPa respectively. The same value of scaffolds crosslinked by 1.2mol/l EDAC were 1.49 and 532.50kPa. Scaffolds crosslinked by 1.0% and 2.5% GTA were toxic to cells, while EDAC groups showed no cytotoxicity. Chondrocytes could proliferate and infiltrate within scaffolds after seeding. Overall, 1.2mol/l EDAC was a preferable crosslinking condition.

Additively Manufactured Macroporous Titanium with Silver-Releasing Micro-/Nanoporous Surface for Multipurpose Infection Control and Bone Repair - A Proof of Concept.

Restoring large-scale bone defects, where osteogenesis is slow while infections lurk, with biomaterials represents a formidable challenge in orthopedic clinics. Here, we propose a scaffold-based multipurpose anti-infection and bone repairing strategy to meet such restorative needs. To do this, personalized multifunctional titanium meshes were produced through an advanced additive manufacturing process and dual "TiO2-poly(dopamine)/Ag (nano)" post modifications, yielding macroporous constructs with micro-/nanoporous walls and nanosilver bullets immobilized/embedded therein. Ultrahigh loading capacity and durable release of Ag+ were accomplished. The scaffolds were active against planktonic/adherent bacteria (Gram-negative and positive) for up to 12 weeks. Additionally, they not only defended themselves from biofilm colonization but also helped destroy existing biofilms, especially in combination with antibiotics. Further, the osteoblasts/bacteria coculture study displayed that the engineered surfaces aided MG-63 cells to combat bacterial invasion. Meanwhile, the scaffolds elicited generally acceptable biocompatibility (cell adhesion, proliferation, and viability) and hastened osteoblast differentiation and maturation (alkaline phosphatase production, matrix secretion, and calcification), by synergy of micro-/nanoscale topological cues and bioactive catecholamine chemistry. Although done ex vivo, these studies reveal that our three-in-one strategy (infection prophylaxis, infection fighting, and bone repair) has great potential to simultaneously prevent/combat infections and bridge defected bone. This work provides new thoughts to the use of enabling technologies to design biomaterials that resolve unmet clinical needs.

Proteomic profile of mouse fibroblasts exposed to pure magnesium extract.

Magnesium and its alloys gain wide attention as degradable biomaterials. In order to reveal the molecular mechanism of the influence of biodegradable magnesium on cells, proteomics analysis was performed in this work. After mouse fibroblasts (L929) were cultured with or without Mg degradation products (Mg-extract) for 8, 24, and 48h, changes in protein expression profiles were obtained using isobaric tags for relative and absolute quantitation (iTRAQ) coupled two dimensional liquid chromatography-tandem mass spectrometry (2D LC MS/MS). A total of 867 proteins were identified (relying on at least two peptides). Compared to the control group, 205, 282, and 217 regulated proteins were identified at 8, 24, and 48h, respectively. 65 common proteins were up or down- regulated within all the three time points, which were involved in various physiological and metabolic activities. Consistent with viability, proliferation, and cell cycle analysis, stimulated energy metabolism as well as protein synthesis pathways were discussed, indicating a possible effect of Mg-extract on L929 proliferation. Furthermore, endocytosis and focal adhesion processes were also discussed. This proteomics study uncovers early cellular mechanisms triggered by Mg degradation products and highlights the cytocompatibility of biodegradable metallic materials for biomedical applications such as stents or orthopaedic implants.

How Microelectrode Array-Based Chick Forebrain Neuron Biosensors Respond to Glutamate NMDA Receptor Antagonist AP5 and GABAA Receptor Antagonist Musimol.

We have established a long-term, stable primary chick forebrain neuron (FBN) culture on a microelectrode array platform as a biosensor system for neurotoxicant screening and for neuroelectrophysiological studies for multiple purposes. This paper reports some of our results, which characterize the biosensor pharmacologically. Dose-response experiments were conducted using NMDA receptor antagonist AP5 and GABAA receptor agonist musimol (MUS). The chick FBN biosensor (C-FBN-biosensor) responds to the two agents in a pattern similar to that of rodent counterparts; the estimated EC50s (the effective concentration that causes 50% inhibition of the maximal effect) are 2.3 µM and 0.25 µM, respectively. Intercultural and intracultural reproducibility and long-term reusability of the C-FBN-biosensor are addressed and discussed. A phenomenon of sensitization of the biosensor that accompanies intracultural reproducibility in paired dose-response experiments for the same agent (AP5 or MUS) is reported. The potential application of the C-FBN-biosensor as an alternative to rodent biosensors in shared sensing domains (NMDA receptor and GABAA receptor) is suggested.

Enhanced in Vitro and in Vivo Performance of Mg-Zn-Y-Nd Alloy Achieved with APTES Pretreatment for Drug-Eluting Vascular Stent Application.

Bioabsorbable magnesium alloys are becoming prominent as temporary functional implants, as they avoid the risks generated by permanent metallic implants such as persistent inflammation and late restenosis. Nevertheless, the overfast corrosion of Mg alloys under physiological conditions hinders their wider application as medical implant materials. Here we investigate a simple one-step process to introduce a cross-linked 3-amino-propyltrimethoxysilane (APTES) silane physical barrier layer on the surface of Mg-Zn-Y-Nd alloys prior to electrostatic spraying with rapamycin-eluting poly(lactic-co-glycolic acid) (PLGA) layer. Surface microstructure was characterized by scanning electron microscope and Fourier transform infrared spectroscopy. Nanoscratch test verified the superior adhesion strength of PLGA coating in the group pretreated with APTES. Electrochemical tests combined with long-term immersion results suggested that the preferable in vitro anticorrosion behavior could be achieved by dense APTES barrier. Cell morphology and proliferation data demonstrated that APTES pretreated group resulted in remarkably preferable compatibility for both human umbilical vein endothelial cells and vascular smooth muscle cells. On the basis of excellent in vitro mechenical property, the animal study on the APTES pretreated Mg-Zn-Y-Nd stent implanted into porcine coronary arteries confirmed benign tissue compatibility as well as re-endothelialization without thrombogenesis or in-stent restenosis at six-month followup.

From Solution to Biointerface: Graphene Self-Assemblies of Varying Lateral Sizes and Surface Properties for Biofilm Control and Osteodifferentiation.

Bringing multifunctional graphene out of solution through facile self-assembly to form 2D surface nanostructures, with control over the lateral size and surface properties, would be an intriguing accomplishment, especially in biomedical fields where biointerfaces with functional diversity are in high demand. Guided by this goal, in this work, we built such graphene-based self-assemblies on orthopedic titanium, attempting to selectively regulate bacterial activities and osteoblastic functions, which are both crucial in bone regeneration. Briefly, large-area graphene oxide (GO) sheets and functionalized reduced GO (rGO) micro-/nanosheets were self-assembled spontaneously and controllably onto solid Ti, through an evaporation-assisted electrostatic assembly process and a mussel-inspired one-pot assembly process, respectively. The resultant layers were characterized in terms of topological structure, chemical composition, hydrophilicity, and protein adsorption properties. The antibacterial efficacies of the assemblies were examined by challenging them with pathogenic Staphylococcus aureus (S. aureus) bacteria that produce biofilms, whereby around 50% antiadhesion effects and considerable antibiofilm activities were observed for both layer types but through dissimilar modes of action. Their cytocompatibility and osteogenic potential were also investigated. Interfaced with MC3T3-E1 cells, the functionalized rGO sheets evoked better cell adhesion and growth than GO sheets, whereas the latter elicited higher osteodifferentiation activity throughout a 28-day in vitro culture. In this work, we showed that it is technically possible to construct graphene interface layers of varying lateral dimensions and surface properties and confirmed the concept of using the obtained assemblies to address the two major challenges facing orthopedic clinics. In addition, we determined fundamental implications for understanding the surface-biology relationship of graphene biomaterials, in efforts to better design and more safely use them for future biomedicine.

Microelectrode Array-evaluation of Neurotoxic Effects of Magnesium as an Implantable Biomaterial.

Magnesium (Mg)-based biomaterials have shown great potential in clinical applications. However, the cytotoxic effects of excessive Mg2+ and the corrosion products from Mg-based biomaterials, particularly their effects on neurons, have been little studied. Although viability tests are most commonly used, a functional evaluation is critically needed. Here, both methyl thiazolyl tetrazolium (MTT) and lactate dehydrogenase (LDH) assays were used to test the effect of Mg2+ and Mg-extract solution on neuronal viability. Microelectrode arrays (MEAs), which provide long-term, real-time recording of extracellular electrophysiological signals of in vitro neuronal networks, were used to test for toxic effects. The minimum effective concentrations (ECmin) of Mg2+ from the MTT and LDH assays were 3 mmol/L and 100 mmol/L, respectively, while the ECmin obtained from the MEA assay was 0.1 mmol/L. MEA data revealed significant loss of neuronal network activity when the culture was exposed to 25% Mg-extract solution, a concentration that did not affect neuronal viability. For evaluating the biocompatibility of Mg-based biomaterials with neurons, MEA electrophysiological testing is a more precise method than basic cell-viability testing.