Corrosion and in vitro cytocompatibility investigation on the designed Mg-Zn-Ag metallic glasses for biomedical application

In the present work,seven Mg-Zn-Ag alloys with the nominal composition of Mg_(96-x)Zn_(x)Ag_(4)(x=17,20,23,26,29,32,35 in at.%)were prepared by induction melting and single-roller melt-spinning.The X-ray diffraction(XRD)analyses indicate the metallic glasses with three composition of Mg_(73)Zn_(23)Ag_(4),Mg_(70)Zn_(26)Ag_(4),and Mg_(67)Zn_(29)Ag_(4)were obtained successfully.The differential scanning calorimetry(DSC)measurement was used to obtain the characteristic temperature of Mg-Zn-Ag metallic glasses for the glass-forming ability analysis.The maximum glass transition temperature(Trg)was found to be 0.525 with a composition close to Mg_(67)Zn_(29)Ag_(4),which results in the best glass-forming ability.Moreover,the immersion test in simulated body fluid(SBF)demonstrate the relative homogeneous corrosion behavior of the Mg-Zn-Ag metallic glasses.The corrosion rate of Mg-Zn-Ag metallic glasses in SBF solution decreases with the increase of Zn content.The sample Mg_(67)Zn_(29)Ag_(4)has the lowest corrosion rate of 0.19mm/yr,which could meet the clinical application requirement well.The in vitro cell experiments show that the Madin-Darby canine kidney(MDCK)cells cultured in sample Mg_(67)Zn_(29)Ag_(4)and its extraction medium have higher activity.However,the Mg-Zn-Ag metallic glasses exhibit obvious inhibitory effect on human rhabdomyosarcoma(RD)tumor cells.The present investigations on the glass-forming ability,corrosion behavior,cytocompatibility and tumor inhibition function of the Mg-Zn-Ag based metallic glass could reveal their biomedical application possibility.

Photo-responsive Carboxymethyl Chitosan/Laponite Hydrogel as a Potential Spinal Cord Injury Scaffold:Characterization and Cytocompatibility Study

We synthesized photo-responsive carboxymethyl chitosan(CMC-MA)via free radical polymerization and utilized nanoclay laponite(LAP)as an inorganic crosslinking agent to develop an injectable and 3D-printable CMC-MA/LAP hydrogel.We determined the optimal ratio of 2.5 w/v%CMC-MA/7.5 w/v%LAP based on injection molding,compression modulus,swelling properties,rheological properties,and 3D printing properties of the hydrogel system.In-vitro cytocompatibility experiments showed that both CMC-MA and CMC-MA/LAP hydrogel had no inhibitory effect on cell proliferation and can promote cell growth when cultured on the surface of the hydrogel matrix.Moreover,the hydrogel containing LAP particles significantly facilitated cell adhesion(>60%)compared with the hydrogel without LAP(20%).Our findings demonstrate that the CMC-MA/LAP hydrogel has great potential for tissue repair in neural tissue engineering.

Biocorrosion property and cytocompatibility of calcium phosphate coated Mg alloy

Calcium phosphate coated Mg alloy was prepared. The phase constitute and surface morphology were identified and observed by X-ray diffractometer （XRD） and SEM. The results show that the coating is composed of flake-like CaHPO4-2H2O crystals. The corrosion resistance of the coated Mg alloy was measured by electrochemical polarization and immersion test in comparison with uncoated Mg alloy. Cytocompatibility was designed by observing the attachment, growth and proliferation of L929 cell on both coated and uncoated Mg alloy samples. The results display that the corrosion resistance of the coated Mg alloy is better than that of uncoated one. The immersion test also shows that the calcium phosphate coating can mitigate the corrosion of Mg alloy substrate, and tends to transform into hydroxyapatite （HA）. Compared with uncoated Mg alloy, L929 cells exhibit good adherence, growth and proliferation characteristics on the coated Mg alloy, indicating that the cytocompatibility is significantly improved with the calcium phosphate coating.

Cytocompatibility with osteogenic cells and enhanced in vivo anti-infection potential of quaternized chitosan-loaded titania nanotubes

Infection is one of the major causes of failure of orthopedic implants. Our previous study demonstrated that nanotube modification of the implant surface, together with nanotubes loaded with quaternized chitosan （hydroxypropyltrimethyl ammonium chloride chitosan, HACC）, could effectively inhibit bacterial adherence and biofilm formation in vitro. Therefore, the aim of this study was to further investigate the in vitro cytocompatibility with osteogenic cells and the in vivo anti-infection activity of titanium implants with HACC-loaded nanotubes （NT-H）. The titanium implant （Ti）, nanotubes without polymer loading （NT）, and nanotubes loaded with chitosan （NT-C） were fabricated and served as controls. Firstly, we evaluated the cytocompatibility of these specimens with human bone marrow-derived mesenchymal stem cells in vitro. The observation of cell attachment, proliferation, spreading, and viability in vitro showed that NT-H has improved osteogenic activity compared with Ti and NT-C. A prophylaxis rat model with implantation in the femoral medullary cavity and inoculation with methiciUin-resistant Staphylococcus aureus was established and evaluated by radiographical, microbiological, and histopathological assessments. Our in vivo study demonstrated that NT-H coatings exhibited significant anti-infection capability compared with the Ti and NT-C groups. In conclusion, HACC-loaded nanotubes fabricated on a titanium substrate show good compatibility with osteogenic cells and enhanced anti-infection ability in vivo, providing a good foundation for clinical application to combat orthopedic implant-associated infections.

Superhydrophobic fluoride conversion coating on bioresorbable magnesium alloy——fabrication,characterization,degradation and cytocompatibility with BMSCs

A micro-nano structure CaF_(2)chemical conversion layer was prepared on fluoride-treated AZ31 alloy,then the composite fluoride conversion film(CaF_(2)/MgF_(2))was modified by stearic acid(SA)and fabricated a superhydrophobic surface.The fluoride-treated magnesium,fluoride conversion film and superhydrophobic coating were characterized by SEM,EDS,XRD and FTIR.The properties of coatings1 adhesion and corrosion resistance were evaluated via tape test and electrochemical measurement.The cytocompatibility of the MgF_(2),CaF_(2)and superhydrophobic CaF_(2)/SA surface was investigated with bone marrow-derived mesenchymal stem cells(BMSCs)by direct culture for 24 h.The results showed that the superhydrophobic fluoride conversion coating composed of inner MgF_(2)layer and the outer CaF_(2)/SA composite layer had an average water contact angle of 152°.SA infiltrated into the micro-nano structure CaF_(2)layer and formed a strong adhesion with CaF_(2)layer.Furthermore,the super-hydrophobic coating showed higher barrier properties and corrosion resistance compared with the fluoride conversion film and fluoride-treated AZ31 alloy.The BMSC adhesion test results demonstrated MgF_(2)CaF_(2)and CaF_(2)/SA coatings were all nontoxic to BMSC.At the condition of in direct contact with cells,MgF_(2)showed higher cell density and enhanced the BMSCs proliferation,while CaF_(2)and CaF_(2)/SA coating showed no statistically difference in cell density compared with glass reference but the CaF_(2)and CaF_(2)/SA coating were not conducive to BMSCs adhesion.

In vitro evaluation of degradation,cytocompatibility and antibacterial property of polycaprolactone/hydroxyapatite composite coating on bioresorbable magnesium alloy

Polycaprolactone/hydroxyapatite(PCL/HA)composite coating was fabricated by a combination of hydrothermal and dipping methods to delay the degradation of Mg alloy AZ31 as bioresorbable materials.The PCL/HA coating was composed of nano rod-shape HA crystals and PCL filled in the space of HA crystals.Compared with the single HA coating,the binding strength between the PCL/HA composite coating and Mg alloy was obviously improved and the PCL/HA coating still adhered to the surface of AZ31 substrate even after 38 days of immersion.The electrochemical corrosion rate of HA coated sample was reduced by ten times after being filled by PCL.The electrochemical impedance spectroscopy(EIS)and immersion test results showed that the PCL/HA composite coating could provide a more effective barrier for Mg substrate than the HA coating alone.The cytocompatibility and the antibacterial property of HA coating and PCL/HA coating were evaluated by culturing with bone marrow-derived mesenchymal stem cells(BMSCs)and methicillin-resistant staphylococcus aureus(MRSA)for 24 h under direct culture conditions,respectively.The PCL/HA composite coating showed better BMSC cell compatibility,more suitable for BMSC adhesion than HA coating alone and showed a potential application prospect as a biological materials.However,from the perspective of clinical applications,the antibacterial property of PCL/HA composite coating needs to be further improved.

Promoted Cytocompatibility of Silk Fibroin Fiber Vascular Graft through Chemical Grafting with Bioactive Molecules

Natural silk from Bombyx mori has been used as medical sutures for several decades,and regenerated silk fibroin( RSF)based biomaterials have been increasingly studied in the past thirty years. However,vascular graft derived from silk fibroin fiber has been explored in recent several years with development of textile science and engineering. Moreover,endothelialization of vascular graft has been seen as an ideal strategy for preventing thrombosis and getting higher patency in a long term. Therefore,in the present work silk fibroin fiber vascular graft( SF) was chemically grafted with bioactive molecules such as heparin and RSF to improve the cytocompatibility. 3-aminopropyl-triethoxysilane(APTES),1-ethyl-3-(3-dimethylaminopropyl) carbodiie hydrochlide( EDC · HCl),and N-hydroxysuccinimide( NHS) have been employed as coupling agent and crosslinking agents,respectively. Microscopy and ATRFTIR were used to characterize the surface changes and the structure of the grafts after treatment,respectively. Cell culture in vitro and MTT assay were conducted to determine the improvement of cell affinity to the graft. Furthermore,mechanical properties of the grafts before and after treatment were compared. The results showed that the chemical grafting was an effective method for improving the cytocompatibility of SF without significant loss of mechanical properties.

Preparation,Rheological Properties and Primary Cytocompatibility of TPU/PLA Blends as Biomedical Materials

A polymer blends containing thermoplastic polyurethane（TPU） and poly（lactic acid）（PLA） as a biomedical material were prepared by a process of modifying thermally induced phase separation（MTIPS） and melt blending.The influences of composition,shear frequency,and temperature on the rheological behaviors of the blends were investigated by small amplitude oscillatory shear rheology.The results revealed that the addition of TPU into PLA significantly decreased the non-Newtonian index of the blends,and increased the sensitivity of the blends on shear rate,suggesting that optimization of the shear rate and temperature could improve the flowability of the blend melts in the extrusion process.In addition,the results of SEM images revealed that TPU distributed well into PLA matrix and showed good compatibility between the TPU and PLA,which made the blends with good toughness.The primary cytocompatibility of the blends was evaluated using C2C12 cells.The results suggested that the TPU/PLA blends did not affect cell growth,showing no cytotoxicity.In short,the TPU/PLA blends with excellent toughness had potential application as biomedical devices.

Preparation of Silk Fibroin Microspheres and Its Cytocompatibility

The goal of this proof-of-concept study was the fabrication of porous silk fibroin (SF) microspheres which could be used as cell culture carriers under very mild processing conditions. The SF solution was differentiated into droplets which were induced by a syringe needle in the high-voltage electrostatic field. They were collected and frozen in liquid nitrogen and water in droplets formed ice crystals which sublimated during lyophilization and a great quantity of micropores shaped in SF microspheres. Finally, the microspheres were treated in ethanol so as to transfer the molecular conformation into β-sheet and then they were insoluble in water. SF particles were spherical in shape with diameters in the range of 208.4 μm to 727.3 μm, while the pore size on the surface altered from 0.3 μm to 10.7 μm. In vitro, the performances of SF microspheres were assessed by culturing L-929 fibroblasts cells. Cells were observed to be tightly adhered and fully extended;also a large number of connections were established between cells. After 5-day culture, it could be observed under a confocal laser scanning microscope that the porous microenvironment offered by SF particles accelerated proliferation of cells significantly. Furthermore, porous SF particles with smaller diameters (200 - 300 μm) might promote cell growth better. These new porous SF microspheres hold a great potential for cell culture carriers and issue engineering scaffolds.

Cytocompatibility of Highly Dispersed Nano Hydroxyapatite Sol

Nano hydroxyapatite (HA) crystals were prepared and dispersed in water to form HA sol by simple methods. The cytotoxicity of the sols were tested by MTT assay and lymphocytotoxicity test. Results show that the average secondary particle size of the sol containing uncalcined HA crystals is around 750 nm, within micrograde; while the sol of calcined HA contains over 88% nanoparticles with the size between 65～86 nm, in which nano HA crystals are highly dispersed. Both the HA sols have no toxicity on the proliferation of 3T3 cells and lymphocytes. It demonstrates that the nano sol is safe for the application of drug delivery.

Polyurethane/poly(vinyl alcohol) hydrogel coating improves the cytocompatibility of neural electrodes

Neural electrodes,the core component of neural prostheses,are usually encapsulated in polydimethylsiloxane（PDMS）.However,PDMS can generate a tissue response after implantation.Based on the physicochemical properties and excellent biocompatibility of polyurethane（PU）and poly（vinyl alcohol）（PVA）when used as coating materials,we synthesized PU/PVA hydrogel coatings and coated the surface of PDMS using plasma treatment,and the cytocompatibility to rat pheochromocytoma（PC12）cells was assessed.Protein adsorption tests indicated that the amount of protein adsorption onto the PDMS substrate was reduced by 92%after coating with the hydrogel.Moreover,the PC12 cells on the PU/PVA-coated PDMS showed higher cell density and longer and more numerous neurites than those on the uncoated PDMS.These results indicate that the PU/PVA hydrogel is cytocompatible and a promising coating material for neural electrodes to improve their biocompatibility.

Cytocompatibility of Self-assembled Hydrogel from IKVAV-containing Peptide Amphiphile with Neural Stem Cells

Neural Stem Cells （NSCs） were incubated with self-assembled hydrogel from IKVAV-containing peptide amphiphile （IKVAV-PA） for one week. The cytocompatibility of hydrogel was evaluated. NSCs were seeded in three-dimensional （3D） hydrogels （Experimental Group, EG） or surface of coverslips （Control Group, CG）, double-labeled with Calcein-AM and PI. A growth curve of cells was obtained according to CCK-8. TEM study of hydrogel revealed a network of nanofibers. NSCs began to proliferate after 24 h of incubation, and formed bigger neurospheres at 48 h in EG than in CG. Cell proliferation activity was higher in EG than in CG （P〈0.05）. The self-assembled Hydrogel had good cytocompatibility and promoted the proliferation of NSCs.

Comparison of nickle release and cytocompatibility between porous and dense NiTi alloy

The porous NiTi(pNiTi)samples were produced by sintering evaporation using Ti−50.8Ni(at.%)gasatomized powders.The samples were analyzed by metallographic microscope and X-ray dispersive spectroscopy(XRD).A comparison of nickel(Ni)release and cytocompatibility between pNiTi and dense NiTi(dNiTi)was made.The results showed that the pNiTi has good mechanical properties.Ni releases from pNiTi in vitro and in vivo are more serious than those form dNiTi.The proliferation and differentiation of cells cultured with the pNiTi extracting liquid are significantly worse,and the rate of early apoptosis is higher.In conclusion,pNiTi is mechanically similar to bone,but pNiTi releases more Ni and interferes with cell proliferation and differentiation.A significantly cautious approach should be adopted when using it as a medical implant.

Surface Modification of Polyglycolic Acid Fibers by Hydrogen Peroxide for Enhancing Hydrophilicity and Cytocompatibility

Hydrogen peroxide( H_2O_2) is applied for surface modification of polyglycolic acid( PGA) fibers in order to enhance the hydrophilicity and cytocompatibility of PGA fibers effectively,and maintain the breaking strength as the same time. PGA fibers are dipped in H_2O_2 solution a certain time for modification. Scanning electron microscopy( SEM) was used to observe the surface morphology of PGA fibers before and after modification. The varying of PGA macromolecule was examined with Fourier transform infrared spectroscopy( FTIR) analyses. X-ray diffraction( XRD) and differential scanning calorimetry( DSC) analysis showed that crystallinity slightly decreases. Mechanical performance test showed tensile force of modified PGA fiber was increased. The water contact angle test indicated the improving of hydrophilic. A cell proliferation assay showed that fibroblast cells attach and proliferate well on the fibers, which meant the modified fibers possess good cytocompatibility. These results suggest that H_2O_2 surface modification is easy to operate and a advantageous modification method for PGA fibers.

The Cytocompatibility of Genipin-Crosslinked Silk Fibroin Films

There is an increasing demand for crosslinking methods of silk fibroin (SF) scaffolds in biomedical applications that could maintain the biocompatibility, bioactivity as well as improve the water resistance and mechanical properties of SF materials. In this study, SF was crosslinked effectively with genipin which is a naturally occurring iridoid glucoside and the crosslinking mechanism was investigated through FTIR and amino acid analysis. The results showed that genipin could react with the -NH2 groups on the side chains of SF macromolecules and to form inter- and intra-molecular covalent bonds, and improved the stability of SF materials significantly. In vitro, the performances of genipin-crosslinked SF films were assessed by seeding L929 cells and compared with ethanol-processed SF films, glutaraldehyde and polyethylene glycol diglycidyl ether crosslinked ones. The genipin-crosslinked SF films showed a similar affinity to cells as ethanol-processed ones, and a higher bioactivity in promoting cell growth and proliferation, inhibition of cell apoptosis, and maintenance of normal cell cycle compared with glutaraldehyde and polyethylene glycol diglycidyl ether crosslinked SF films. These features, combined with the decrease of brittleness of SF films crosslinked with chemical methods, substantiated genipin as an effective and biocompatible agent for the manufacturing of bioactive SF materials which used as tissue engineering scaffolds and drug delivery carriers.

Cytocompatibility of Three Corneal Cell Types with Amniotic Membrane

Rabbit limbal corneal epithelial cells,corneal endothelial cells and keratocytes were cultured on amniotic membrane. Phase contrast microscope examination was performed daily. Histological and scan electron microscopic examinations were carried out to observe the growth,arrangement and adhesion of cultivated cells. Results showed that three corneal cell types seeded on amniotic membrane grew well and had normal cell morphology. Cultured cells attached firmly on the surface of amniotic membrane. Corneal epithelial cells showed singular layer or stratification. Cell boundaries were formed and tightly opposed. Corneal endothelial cells showed cobblestone or polygonal morphologic characteristics that appeared uniform in size. The cellular arrangement was compact. Keratocytes elongated and showed triangle or dendritic morphology with many intercellular joints which could form networks. In conclusion,amniotic membrane has good scaffold property,diffusion effect and compatibility with corneal cells. The basement membrane side of amniotic membrane facilitated the growth of corneal epithelial cells and endothelial cells and cell junctions were tightly developed. The spongy layer of amniotic membrane facilitated the growth of keratocytes and intercellular joints were rich. Amniotic membrane is an ideal biomaterial for layering tissue engineered cornea.

Optimization of the in vitro biodegradability,cytocompatibility,and wear resistance of the AZ31B alloy by micro-arc oxidation coatings doped with zinc phosphate

As implanted bone fixation materials,magnesium(Mg)alloys have significant advantages because the density and elastic modulus are closest to those of the human bone and they can bio-degrade in the physiological environment.However,Mg alloys degrade too rapidly and uncontrollably thus hampering clinical adoption.In this study,a highly corrosion-resistant zinc-phosphate-doped micro-arc oxidation(MAO)coating is prepared on the AZ31B alloy,and the degradation process is assessed in vitro.With increasing zinc phosphate concentrations,both the corrosion potentials and charge transfer resistance of the AZ31B alloy coated with MAO coatings increase gradually,while the corrosion current densities di-minish gradually.Immersion tests in the simulated body fluid(SBF)reveal that the increased zinc phos-phate concentration in MAO coating decreases the degradation rate,consequently reducing the release rates of Mg^(2+)and OH-in the physiological micro-environment,which obtains the lowest weight loss of only 5.22%after immersion for 56 days.Effective regulation of degradation provides a weak alkaline environment that is suitable for long-term cell growth and subsequent promotion of bone proliferation,differentiation,mineralization,and cytocompatibility.In addition,the zinc-phosphate-doped MAO coat-ings show an improved wear resistance as manifested by a wear rate of only 3.81 x 10^(-5) mm^(3) N^(-1) m^(-1).The results reveal a suitable strategy to improve the properties of biodegradable Mg alloys to balance tissue healing with mechanical degradation.

A homogeneous dopamine-silver nanocomposite coating:striking a balance between the antibacterial ability and cytocompatibility of dental implants

Silver has been widely used for surface modification to prevent implant-associated infections.However,the inherent cytotoxicity of silver greatly limited the scope of its clinical applications.The construction of surfaces with both good antibacterial properties and favorable cytocompatibility still remains a challenge.In this study,a structurally homogeneous dopamine-silver(DA/Ag)nanocomposite was fabricated on the implant surface to balance the antibacterial activity and cytocompatibility of the implant.The results show that the DA/Ag nanocomposites prepared under the acidic conditions(pH=4)on the titanium surface are homogeneous with higher Ag^(+)content,while an obvious core(AgNPs)-shell(PDA)structure is formed under neutral(pH=7)and alkaline conditions(pH=10),and the subsequent heat treatment enhanced the stability of PDA-AgNPs nanocomposite coatings on porous titanium.The antibacterial test,cytotoxicity test,hypodermic implantation and osteogenesis test revealed that the homogeneous PDA-AgNPs nanocomposite coating achieved the balance between the antibacterial ability and cytocompatibility,and had the best outcomes for soft tissue healing and bone formation around the implants.This study provides a facile strategy for preparing silver-loaded surfaces with both good antibacterial effect and favorable cytocompatibility,which is expected to further improve the therapeutic efficacy of silver composite-coated dental implants.

Gadolinium-doped injectable magnesium-calcium phosphate bone cements for noninvasive visualization

Injectable bone cements are used in minimally invasive surgical techniques including vertebroplasty and kyphoplasty.This work is devoted to the development of magnesium-calcium phosphate cements(MCPCs)doped with gadolinium ions(Gd^(3+))for bone defect repair.Interaction between cement powders and a cement liquid resulted in the formation of newberyite and brushite phases,which gave mechanical strength up to 17 MPa without a thermal effect.The introduction of Gd3+into the lattice was confirmed by electron paramagnetic resonance spectroscopy;the doping increased injectivity while giving rise to antibacterial properties against Escherichia coli.Assays of the cement samples soaking in Kokubo’s simulated body fluid revealed the formation of calcium phosphate coatings on the cements’surface.The cements manifested biocompatibility with the MG-63 cell line and significantly enhanced contrast when Gd-MCPC was placed into a bone defect and examined by X-ray micro-computed tomography.For the first time,visualization of a Gd-doped cement material was achieved in a model of a bone defect analyzed by MRI.

Design and characterization of biodegradable Mg-Zn-Ag metallic glasses

In order to develop the Mg-Zn-Ag metallic glasses(MGs)for biodegradable implant applications,the glass formation ability(GFA)and biocompatibility of Mg-Zn-Ag alloys were investigated using a combination of the calculation of phase diagrams(CALPHAD)and experimental measurements.High GFA potentiality of two alloy series,specifically Mg_(96-x)Zn_xAg_(4)and Mg_(94-x)Zn_xAg_6(x=17,20,23,26,29,32,35),was predicted theoretically and then substantiated through experimental testing.X-ray diffraction(XRD)and differential scanning calorimetry(DSC)techniques were used to evaluate the crystallinity,GFA,and crystallization characteristics of these alloys.The results showed that compositions between Mg_(73)Zn_(23)Ag_(4)and Mg_(64)Zn_(32)Ag_(4)for Mg_(96-x)Zn_xAg_4,Mg_(66)Zn_(28)Ag_(6)and Mg_(63)Zn_(31)Ag_(6for)Mg_(94-x)Zn_xAg_(6)displayed a superior GFA.Notably,the GFA of the Mg_(96-x)Zn_xAg_(4)series was better than that of the Mg_(94-x)Zn_xAg_(6)series.Furthermore,the Mg_(70)Zn_(26)Ag_4,Mg_(74)Zn_(20)Ag_6,and Mg_(71)Zn_(23)Ag_(6)alloys showed acceptable corrosion rates,good cytocompatibility,and positive effects on cell proliferation.These characteristics make them suitable for applications in medical settings,potentially materials as biodegradable implants.