PMMA-silica nanocomposite coating: Effective corrosion protection and biocompatibility for a Ti6Al4V alloy.

Ti6Al4V is the mostly applied metallic alloy for orthopedic and dental implants, however, its lack of osseointegration and poor long-term corrosion resistance often leads to a secondary surgical intervention, recovery delay and toxicity to the surrounding tissue. As a potential solution of these issues poly(methyl methacrylate)-silicon dioxide (PMMA-silica) coatings have been applied on a Ti6Al4V alloy to act simultaneously as an anticorrosive barrier and bioactive film. The nanocomposite, composed of PMMA covalently bonded to the silica phase through 3-(trimethoxysilyl)propyl methacrylate (MPTS), has been synthesized combining the sol-gel process with radical polymerization of methyl methacrylate. The 5 µm thick coatings deposited on Ti6Al4V have a smooth surface, are homogeneous, transparent, free of pores and cracks, and show a strong adhesion to the metallic substrate (11.6 MPa). Electrochemical impedance spectroscopy results proved an excellent anticorrosive performance of the coating, with an impedance modulus of 26 GΩ cm2 and long-term durability in simulated body fluid (SBF) solution. Moreover, after 21 days of immersion in SBF, the PMMA-silica coating presented apatite crystal deposits, which suggests in vivo bone bioactivity. This was confirmed by biological characterization showing enhanced osteoblast proliferation, explained by the increased surface free energy and protein adsorption. The obtained results suggest that PMMA-silica hybrids can act in a dual role as efficient anticorrosive and bioactive coating for Ti6Al4V alloys.

Dual effective core-shell electrospun scaffolds: Promoting osteoblast maturation and reducing bacteria activity.

Herein, we electrospun ultrathin core-shell fibers based on polycaprolactone (PCL), polyethylene glycol (PEG), gelatin and osteogenic growth peptide (OGP), and evaluated their potential to upregulate human osteoblast cells (hFOB) and to reduce Gram-positive and Gram-negative bacteria. We also evaluated the fiber morphology, chemical structure and peptide delivery efficacy. The employment of core-shell fibers compared to fibers without a core-shell showed improved mechanical strength, comparable to the strength of pure PCL, as well as improved hydrophilicity and wettability. The careful selection of polymer combination and core-shell strategy promoted a controlled and sustained release of OGP. Moreover, increased calcium deposition (CD) (1.3-fold) and alkaline phosphate (ALP) activity was observed when hFOBs were cultivated onto core-shell fibers loaded with OGP after 21 days of culture. Our developed scaffolds were also able to reduce the amount of Pseudomonas aeruginosa (ATCC 25668) bacteria by a factor of two compared to raw PCL without the use of any antibiotics. All of these results demonstrate a promising potential of the developed core-shell electrospun scaffolds based on PCL:PEG:Gelatin:OGP for numerous bone tissue applications.

Electrospun nanofiber blend with improved mechanical and biological performance.

BACKGROUND: Here, electrospun fibers based on a blend of polycaprolactone (PCL), poly(ethylene glycol) (PEG), and gelatin methacryloyl (GelMA) were developed. The careful choice of this polymer combination allowed for the preparation of a biomaterial that preserved the mechanical strength of PCL, while at the same time improving the hydrophilicity of the blended material and human osteoblast maturation. METHODS: The morphology, chemical structure, wettability, and mechanical properties before and after UV photocrosslinking were evaluated. Furthermore, human osteoblasts (hFOB) were cultivated for up to 21 days on the scaffolds, and their potential to upregulate cell proliferation, alkaline phosphatase (ALP) activity, and calcium deposition were investigated. RESULTS: Contact angle measurement results showed that the developed scaffolds presented hydrophilic properties after PEG and GelMA incorporation before (25°) and after UV photocross-linking (69°) compared to pure PCL (149°). PCL:PEG:GelMA-UV displayed a slight increase in mechanical strength (elastic modulus ~37 MPa) over PCL alone (~33 MPa). Normally, an increase in strength of fibers leads to a decrease in elongation at break, due to the material becoming less deformable and stiffer, thus leading to breaks at low strain. This behavior was observed by comparing PCL (elongation at break ~106%) and PCL:PEG:GelMA-UV (~50%). Moreover, increases in ALP activity (10-fold at day 14) and calcium deposition (1.3-fold at day 21) by hFOBs were detected after PEG and GelMA incorporation after UV photocross-linking compared to pure PCL. Ultrathin and hydrophilic fibers were obtained after PEG and GelMA incorporation after UV photocrosslinking, but the strength of PCL was maintained. Interestingly, those ultrathin fiber characteristics improved hFOB functions. CONCLUSION: These findings appear promising for the use of these electrospun scaffolds, based on the combination of polymers used here for numerous orthopedic applications.

Understanding the impact of crosslinked PCL/PEG/GelMA electrospun nanofibers on bactericidal activity.

Herein, we report the design of electrospun ultrathin fibers based on the combination of three different polymers polycaprolactone (PCL), polyethylene glycol (PEG), and gelatin methacryloyl (GelMA), and their potential bactericidal activity against three different bacteria Staphylococcus aureus (S. aureus), Pseudomonas aeruginosa (P. aeruginosa), and Methicillin-resistant Staphylococcus aureus (MRSA). We evaluated the morphology, chemical structure and wettability before and after UV photocrosslinking of the produced scaffolds. Results showed that the developed scaffolds presented hydrophilic properties after PEG and GelMA incorporation. Moreover, they were able to significantly reduce gram-positive, negative, and MRSA bacteria mainly after UV photocrosslinking (PCL:PEG:GelMa-UV). Furthermore, we performed a series of study for gaining a better mechanistic understanding of the scaffolds bactericidal activity through protein adsorption study and analysis of the reactive oxygen species (ROS) levels. Furthermore, the in vivo subcutaneous implantation performed in rats confirmed the biocompatibility of our designed scaffolds.

Synthesis and study of cell-penetrating peptide-modified gold nanoparticles.

BACKGROUND: In nanomedicine, gold nanoparticles (AuNPs) have demonstrated versatile therapeutic efficiencies and, in particular, have been developed for the treatment of various cancers due to their high selectivity in killing cancer, not healthy, cells. METHODS: In this study, AuNPs were conjugated with the cell-penetrating peptide Cys-(Arg)8-Asp-Ser (CRRRRRRRRGDS) by direct cross-linking of the cysteine's thiol group to the gold surface and a fibronectin-derived RGD group was also used due to its efficacy toward cancer cell targeting and possible promotion of healthy fibroblast functions. RESULTS: Ultraviolet-visible absorbance spectrum and transmission electron microscope images of the synthesized peptide-capped AuNPs (PEP-AuNPs) validated the formation of AuNP aggregates. The presence of peptides on AuNPs was confirmed by Fourier transform infrared spectroscopy and quantified by a bicinchoninic acid assay. After being modified with the arginine-rich peptide, the AuNPs possessed a positive charge, as their zeta potential increased from -23.81±8.43 mV to 8 mV on average. In this manner, an easy method to conjugate AuNPs was shown here. Further, MTS assays were performed using healthy human dermal fibroblasts. After 24 hours of treatment with PEP-AuNPs, the cell density increased dramatically to around 25,000 cells/cm2. Results further showed a very high half-maximal inhibitory concentration of 69.2 µM for the PEP-AuNPs (indicating low toxicity). CONCLUSION: The results showed for the first time the ability of PEP-AuNPs to promote human dermal fibroblast cell viability, which after further investigation, may show an ability to replace cancerous tissue with healthy soft tissue.

Carbon Nanomaterials for Treating Osteoporotic Vertebral Fractures.

PURPOSE OF REVIEW: To identify the use of carbon nanomaterials in bone regeneration and present new data on the regenerative capacity of bone tissue in osteopenic rats treated with graphene nanoribbons (GNRs). RECENT FINDINGS: The results show that the physical and chemical properties of the nanomaterials are suitable for the fabrication of scaffolds intended for bone regeneration. The in vitro tests suggested a non-toxicity of the GNRs as well as improved biocompatibility and bone mineralization activity. Here, for the first time, we evaluated the potential of GNRs in remodeling and repairing bone defects in osteoporotic animal models in vivo. Interestingly, bone mineralization and the initiation of the remodeling cycle by osteoclasts/osteoblasts were observed after the implantation of GNRs, thus implying healthy bone remodeling when using GNRs. This study, therefore, has opened our perspectives and certainly calls for more attention to the use of carbon nanomaterials for a wide range of osteoporosis applications.

Micro-Nanofibrillar Polycaprolactone Scaffolds as Translatable Osteoconductive Grafts for the Treatment of Musculoskeletal Defects without Infection.

The treatment of musculoskeletal defects is currently limited by the tissue-regenerative materials available to orthopedic surgeons: autologous bone grafts only have a finite amount of harvestable material within a given patient, while allografts are prone to severe immunological complications and host rejection. With this motivation, the production of poly(ε-caprolactone) (PCL) scaffolds as synthetic, biomimetic biomaterials was investigated, with a specific focus on potential orthopedic translation. PCL scaffolds were produced through three different fabrication techniques: electrospinning (ES), rotary jet spinning (RJS), and airbrush (AB). ES and RJS were observed to produce microfibrillar scaffolds, while all AB products were nanofibrous. Osteoblast viability, within the PCL scaffolds, and the osteogenic phenotype were assessed in vitro through a combination of adherence, metabolic activity, proliferation, gene expression, alkaline phosphatase bioactivity, and calcium deposition assays. While the polymeric scaffolds induced slight reductions in initial osteoblast adhesion and metabolic activity, seeded cells were able to proliferate and demonstrate the bone formation phenotype. AB products demonstrated reduced bacterial surface colonization when inoculated with both Gram-positive (Staphylococcus aureus) and Gram-negative (Pseudomonas aeruginosa) bacterial strains, in comparison to the microfibrous ES and RJS products, without any small-molecule antibiotics, antimicrobial peptides, or reactive nanomaterials included during scaffold synthesis.

Shape and surface chemistry effects on the cytotoxicity and cellular uptake of metallic nanorods and nanospheres.

Metallic nanoparticles (such as gold and silver) have been intensely studied for wound healing applications due to their ability to be easily functionalized, possess antibacterial properties, and their strong potential for targeted drug release. In this study, rod-shaped silver nanorods (AgNRs) and gold nanorods (AuNRs) were fabricated by electron beam physical vapor deposition (EBPVD), and their cytotoxicity toward human skin fibroblasts were assessed and compared to sphere-shaped silver nanospheres (AgNSs) and gold nanospheres (AuNSs). Results showed that the 39.94 nm AgNSs showed the greatest toxicity with fibroblast cells followed by the 61.06 nm AuNSs, ∼556 nm × 47 nm (11.8:1 aspect ratio) AgNRs, and the ∼534 nm × 65 nm (8.2:1 aspect ratio) AuNRs demonstrated the least amount of toxicity. The calculated IC50 (50% inhibitory concentration) value for the AgNRs exposed to fibroblasts was greater after 4 days of exposure (387.3 µg mL(-1)) compared to the AgNSs and AuNSs (4.3 and 23.4 µg mL(-1), respectively), indicating that these spherical metallic nanoparticles displayed a greater toxicity to fibroblast cells. The IC50 value could not be measured for the AuNRs due to an incomplete dose response curve. The reduced cell toxicity with the presently developed rod-shaped nanoparticles suggests that they may be promising materials for use in numerous biomedical applications.

Shape and surface effects on the cytotoxicity of nanoparticles: Gold nanospheres versus gold nanostars.

Gold nanoparticles are materials with unique optical properties that have made them very attractive for numerous biomedical applications. With the increasing discovery of techniques to synthesize novel nanoparticles such as star-shaped gold nanoparticles for biomedical applications, the safety and performance of these new nanomaterials must be systematically assessed before use. In this study, gold nanostars (AuNSTs) with multibranched surface structures were synthesized, and their influence on the cytotoxicity of human skin fibroblasts and rat fat pad endothelial cells (RFPECs) were assessed and compared with that of gold nanospheres (AuNSPs) with unbranched surfaces. Results showed that the AuNSPs with diameters of approximately 61.46 nm showed greater toxicity with fibroblast cells and RFPECs compared with the synthesized AuNSTs with diameters of approximately 33.69 nm. The AuNSPs were lethal at concentrations of 40 µg/mL for both cell lines, whereas the AuNSTs were less toxic at higher concentrations (400 µg/mL). The calculated IC50 (50% inhibitory concentration) values of the AuNSPs exposed to fibroblast cells were greater at 1 and 4 days of culture (26.4 and 27.7 µg/mL, respectively) compared with the RFPECs (13.6 and 13.8 µg/mL, respectively), indicating that the AuNSPs have a greater toxicity to endothelial cells. It was proposed that possible factors that could be promoting the reduced toxicity effects of the AuNSTs to fibroblast cells and RFPECs, compared with the AuNSPs may be size, surface chemistry, and shape of the gold nanoparticles. The reduced cell toxicity observed with the AuNSTs suggests that AuNSTs may be a promising material for use in biomedical applications.

Nanomedicine for implants: a review of studies and necessary experimental tools.

The response of host organisms (including at the protein and cellular level) to nanomaterials is different than that observed to conventional materials. Nanomaterials are those materials which possess constituents less than 100 nm in at least one direction. This review will first introduce the use of nanomaterials in a variety of implant applications highlighting their promise towards regenerating tissues. Such reviewed studies will emphasize interactions of nanomaterials with various proteins and subsequently cells. Moreover, such advances in the use of nanomaterials as novel implants have been largely, to date, determined by conventional methods. However, the novel structure­property relationships unique for nanosized materials reside at the nanoscale. That is, the novelty of a nanomaterial can only be fully appreciated by characterizing their interactions with biological systems (such as proteins) with nanoscale resolution analytical tools. This characterization of nanomaterials at the nanoscale is critical to understanding and, hence, further promoting increased tissue growth on nanomaterials. For this reason, while more tools are needed for this emerging field, this review will also cover currently available surface characterization techniques that emphasize nanoscale resolution pertinent for characterizing biological interactions with nanomaterials, including attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy, X-ray photoelectron spectroscopy (XPS), time-of-flight secondary ion mass spectroscopy (SIMS), colorimetric biological assays, circular dichroism (CD), and atomic force microscopy (AFM). Only through the coordination of nanoscale analytical tools with studies that highlight mechanisms of increased tissue growth on nanomaterials will we be able to design better implant materials.

The effect of nanotopography on calcium and phosphorus deposition on metallic materials in vitro.

To date, long-term functions of osteoblasts leading to calcium and phosphorus mineral deposition on nanometals have not been determined. Nanometals are metals with constituent metal particles and/or surface features less than 100 nm in at least one dimension. For this reason, the objective of this in vitro study was to determine the amount of calcium and phosphorus mineral formation on microphase compared to nanophase Ti, Ti6Al4V, and CoCrMo cultured with and without osteoblasts (bone-forming cells). The results of this study provided the first evidence of significantly greater calcium and phosphorus deposition by osteoblasts and precipitation from culture media without osteoblasts on nanophase compared to respective microphase Ti6Al4V and CoCrMo after 21 days; the greatest calcium and phosphorus mineral deposition occurred on nanophase CoCrMo while the greatest calcium and phosphorus mineral precipitation without osteoblasts occurred on nanophase Ti6Al4V. No differences were found for any type of Ti: wrought, microphase, or nanophase. Moreover, increased calcium and phosphorus mineral content correlated to greater amounts of underlying aluminum content on Ti6Al4V surfaces. Since, compared to microphase Ti6Al4V, nanophase Ti6Al4V contained a higher amount of aluminum at the surface (due to greater surface area), this may provide a reason for enhanced calcium and phosphorus mineral content on nanophase Ti6Al4V. Regardless of the mechanism, this study continues to support the further investigation of nanometals for improved orthopedic applications.

Increased osteoblast functions on undoped and yttrium-doped nanocrystalline hydroxyapatite coatings on titanium.

In order to improve orthopedic implant performance, the objective of this in vitro study was to synthesize nanocrystalline hydroxyapatite (HA) powders to coat titanium. HA was synthesized through a wet chemical process. The precipitated powders were either sintered at 1100 degrees C for 1h in order to produce UltraCap HA (or microcrystalline size HA) or were treated hydrothermally at 200 degrees C for 20 h to produce nanocrystalline HA. Some of the UltraCap and nanocrystalline HA powders were doped with yttrium (Y) since previous studies demonstrated that Y-doped HA in bulk improved osteoblast (or bone-forming cell) function over undoped HA. The original HA particles were characterized using X-ray diffraction (XRD), inductively coupled plasma-atomic emission spectroscopy (ICP-AES), BET, a laser particle size analyzer, and scanning electron microscopy (SEM). These powders were then deposited onto titanium by a novel room-temperature process, called IonTite. The properties of the resulting HA-coatings on titanium were compared to respective properties of the original HA powders. The results showed that the chemical and physical properties of the original HA powders were retained when coated on titanium by IonTite, as determined by XRD, SEM, and atomic force microscopy (AFM) analysis. More importantly, results showed increased osteoblast adhesion on the nanocrystalline HA IonTite coatings compared to traditionally used plasma-sprayed HA coatings. Results also demonstrated greater amounts of calcium deposition by osteoblasts cultured on Y-doped nanocrystalline HA coatings compared to either UltraCap IonTite coatings or plasma-sprayed HA coatings. These results encourage further studies on nanocrystalline IonTite HA coatings on titanium for improved orthopedic applications.