Multifunctional bioactive glass scaffolds coated with layers of poly(d,l-lactide-co-glycolide) and poly(n-isopropylacrylamide-co-acrylic acid) microgels loaded with vancomycin

A new family of multifunctional scaffolds, incorporating selected biopolymer coatings on basic Bioglass® derived foams has been developed. The polymer coatings were investigated as carrier of vancomycin which is a suitable drug to impart antibiotic function to the scaffolds. It has been proved that coating with PLGA (poly(lactic-co-glycolic acid)) with dispersed vancomycin-loaded microgels provides a rapid delivery of drug to give antibacterial effects at the wound site and a further sustained release to aid mid to long-term healing. Furthermore, the microgels also improved the bioactivity of the scaffolds by acting as nucleation sites for the formation of HA crystals in simulated body fluid.     

Preparation and in vitro bioactivity of poly(d,l-lactide) composite containing hydroxyapatite nanocrystals

The nano-sized hydroxyapatite (n-HA) was incorporated into poly(d,l-Lactide) (PDLLA) to form a bioactive and biodegradable composite for application in hard tissue replacement and regeneration. Thin film of PDLLA composite containing 20 mass% of n-HA fillers was successfully developed through integration of solvent co-blending and hot pressing techniques. firstly, n-HA and PDLLA were chemically synthesized, respectively, then mixed together and homogeneously dispersed in N,N-dimethyl formamide(DMF) solvent, finally, the dried blended hybrid containing PDLLA matrix and n-HA fillers was put into the mould and compacted by hot-pressing machine under 8 MPa pressure at 110 °C for 15 min. In vitro studies were conducted using the simulated body fluid(SBF). Composite specimens were soaked in SBF from 1 day to 21 days prior to surface analysis. Results obtained from scanning electron microscopy(SEM) examination, Energy dispersive X-ray detector(EDX) analysis and X-ray diffraction (XRD) analysis showed that a layer of non-stoichiometric apatite formed within 7 days on HA/PDLLA composite surface after its immersion in SBF, demonstrating moderate in vitro bioactivity of n-HA/PDLLA composite, though a moderate rate of apatite formation in SBF was found on initial stage of immersion periods for n-HA/PDLLA composite, compared to the other biomaterial composite. This type of composite film exhibited certain desirable bioactive characteristics, and they are promising bone candidates to develop novel bioactive composites for biomedical application.     

Osteogenic and antimicrobial nanoparticulate calcium phosphate and poly-(d,l-lactide-co-glycolide) powders for the treatment of osteomyelitis

Development of a material for simultaneous sustained and localized delivery of antibiotics and induction of spontaneous regeneration of hard tissues affected by osteomyelitis stands for an important clinical need. In this work, a comparative analysis of the bacterial and osteoblastic cell response to two different nanoparticulate carriers of clindamycin, an antibiotic commonly prescribed in the treatment of bone infection, one composed of calcium phosphate and the other comprising poly-(D,L-lactide-co-glycolide)-coated calcium phosphate, was carried out. Three different non-cytotoxic phases of calcium phosphate, exhibiting dissolution and drug release profiles in the range of one week to two months to one year, respectively, were included in the analysis: monetite, amorphous calcium phosphate and hydroxyapatite. Spherical morphologies and narrow size distribution of both types of nanopowders were confirmed in transmission and scanning electron microscopic analyses. The antibiotic-containing powders exhibited sustained drug release contingent upon the degradation rate of the carrier. Assessment of the antibacterial performance of the antibiotic-encapsulated powders against Staphylococcus aureus, the most common pathogen isolated from infected bone, yielded satisfactory results both in broths and on blood agar plates for all the analyzed powders. In contrast, no cytotoxic behavior was detected upon the incubation of the antibiotic powders with the osteoblastic MC3T3-E1 cell line for up to three weeks. The cells were shown to engage in a close contact with the antibiotic-containing particles, irrespective of their internal or surface phase composition, polymeric or mineral. At the same time, both types of particles upregulated the expression of osteogenic markers osteocalcin, osteopontin, Runx2 and protocollagen type I, suggesting their ability to promote osteogenesis and enhance remineralization of the infected site in addition to eliminating the bacterial source of infection.     

Methotrexate-immobilized poly(ethylene glycol) magnetic nanoparticles for MR imaging and drug delivery

We report the development of a biostable methotrexate-immobilized iron oxide nanoparticle drug carrier that may potentially be used for real-time monitoring of drug delivery through magnetic resonance imaging. Methotrexate (MTX) was immobilized on the nanoparticle surface via a poly(ethylene glycol) self-assembled monolayer (PEG SAM). The cytotoxicity of the nanoparticle-drug conjugate (NP-PEG-MTX) to target cells was studied with 9L glioma cells. Cellular uptake experiments showed that the uptake of NP-PEG-MTX conjugates by glioma cells was considerably higher than that of control nanoparticles. Magnetic resonance imaging in 9L cells cultured with NP-PEG-MTX of various concentrations showed significant contrast enhancement. NP-PEG-MTX demonstrated higher cytotoxicity in 9L cells to free MTX in vitro. Leucovorin, an MTX antidote, was used to rescue the cells that had been exposed to NP-PEG-MTX or free MTX, and the experiment verified the biocompatibility of NP-PEG-MTX conjugates and the MTX on NP-PEG-MTX conjugates to be the true source of the cytotoxicity to the target cells. TEM results showed that NP-PEG-MTX conjugates were internalized into the 9L cellular cytoplasm and retained its crystal structure therein for up to 144 h, as identified by electron diffraction. This prolonged particle retention may allow physicians to image tumor cells exposed to the NP-PEG-MTX conjugate over an extended therapeutic time course.     

Synthesis and properties of poly(d,l-lactide) drug carrier with maghemite nanoparticles

A magnetic polymeric carrier, composed of biodegradable poly(d,l-lactide) microspheres, maghemite nanoparticles and anti-cancer drug (paclitaxel) was successfully prepared in dichloromethane using high-speed homogenization. Maghemite nanoparticles were well dispersed in PLA matrix. The carrier was magnetically responsive and exhibited low cytotoxicity. Release of loaded paclitaxel was enhanced by applying an oscillating magnetic field. The thermal energy generated by maghemite nanoparticles due to magnetic hysteresis loss was very low and considered negligible in influencing the release behavior. Alternating movement of the nanoparticles, stimulated by magnetic force, resulting in deterioration of the mechanical properties of polymer matrix was likely to be the main reason for the enhancement in drug release.     

Preparation and characterization of novel CdSe quantum dots modified with poly (d,l-lactide) nanoparticles

In this paper, novel CdSe quantum dots (QDs) modified with poly (d, l-lactide) (PLA) nanoparticles by nanoprecipitation method was reported. CdSe QDs modified with PLA nanoparticles were characterized by Photon correlation spectroscope (PCS), transmission electron microscope (TEM), flluorescence spectrophotometer and fluorescence microscope. The modified CdSe QDs were spherical and relatively uniform. The modified CdSe QDs were water soluble and their strong yellow fluorescence emission was observed both in vitro and in vivo. The fluorescence of the modified CdSe QDs was stable in aqueous solution for more than 30 d. These modified CdSe QDs are expected to have much potential for biological labeling and diagnostics based on above properties.     

Novel antibacterial electrospun mats based on poly(d,l-lactide) nanofibers and zinc oxide nanoparticles

This investigation addresses the morphological, mechanical, and antibacterial evaluation of nanocomposite mats based on poly(d,l-lactide) nanofibers with different zinc oxide nanoparticles (nano-ZnO) concentration, that were elaborated by two techniques, i.e., electrospinning of polymer/ZnO solutions and the combination of electrospinning of polymer solutions with electrospraying of nano-ZnO dispersions. The analysis of the precursory solutions was carried out in order to understand the achieved morphology of nanofibers. The obtained poly(d,l-lactide)/ZnO fibrous mats showed a uniform morphology with an average porosity ca. 55 % and average pore size around 45 μm. The presence of ZnO nanoparticles increased the toughness of the mats, and an optimal nano-ZnO concentration (i.e., 3 wt%) was observed at which the tensile strength and Young’s modulus could be improved. Concerning to the antibacterial properties, a relatively low concentration of nanoparticles provoked a growth inhibition of the Gram-negative Escherichia coli and the Gram-positive Staphylococcus aureus bacteria. The mats have potential features for use as antimicrobial wound dressings.     

Facile one-pot synthesis of iron oxide nanoparticles cross-linked magnetic poly(vinyl alcohol) gel beads for drug delivery

In this paper, a facile one-pot strategy for scalable synthesis of robust magnetic poly(vinyl alcohol) (mPVA) gel beads is developed. Through dropwise addition of mixed aqueous solution of iron salts and PVA solution into alkaline (e.g., ammonia, NaOH, and KOH) solution, mPVA gel beads with uniform size and excellent superparamagnetic property can be fabricated based on the simultaneous formation of magnetic iron oxide nanoparticles (MIONs) and cross-link of PVA chains. Moreover, this approach can be extended to prepare dual- or multiresponsive gel beads through simply adding functional fillers into PVA solution (e.g., mPVA-PNIPAM gel beads that possess both magnetic and temperature responsibilities can be readily prepared by adding temperature responsive poly(N-isopropylacrylamide) (PNIPAM) into PVA solution). It is found that that the obtained mPVA gel beads exhibit high drug loading level (e.g., above 70%) after the treatment of freezing-thawing. Drug release experiments reveal that the drug release rate and amount of the mPVA gel beads can be tuned by operating the external magnetic field and adjusting the concentration of iron oxide nanoparticles and temperature (for mPVA-PNIPAM gel beads). The present work is of interest for opening up enormous opportunities to make full use of magnetic gel beads in drug delivery and other applications, because of their facile availability, cost-effective productivity, and tunable drug release performance.     

Size-tunable nanoparticles composed of dextran-b-poly(D,L-lactide) for drug delivery applications

Nanoparticles (NPs) formulated using self-assembly of block copolymers have attracted significant attention as nano-scaled drug delivery vehicles. Here we report the development of a biodegradable NP using self-assembly of a linear amphiphilic block copolymer, Dex-b-PLA, composed of poly(D,L-lactide), and dextran. The size of the NPs can be precisely tuned between 15 and 70 nm by altering the molecular weight (M _W) of the two polymer chains. Using doxorubicin as a model drug, we demonstrated that the NPs can carry up to 21% (w/w) of the drug payload. The release profile of doxorubicin from NPs showed sustained release for over 6 days. Using a rat model, we explored the pharmacokinetics profiles of Dex-b-PLA NPs, and showed proof-of-concept that long circulation lifetime of the NPs can be achieved by tuning the M _W of Dex-b-PLA block copolymer. While the terminal half-life of Dex-b-PLA NPs (29.8 h) was similar to that observed in poly(ethylene glycol)-coated (PEG-coated) NPs (27.0 h), 90% of the injected Dex-b-PLA NPs were retained in the blood circulation for 38.3 h after injection, almost eight times longer than the PEG-coated NPs. The area under curve (AUC) of Dex-b-PLA NPs was almost four times higher than PEG-based NPs. The biodistribution study showed lower accumulation of Dex-b-PLA NPs in the spleen with 19.5% initial dose per gram tissue (IDGT) after 24 h compared to PEG-coated poly(lactide-co-glycolide) (PLGA) NPs (29.8% IDGT). These studies show that Dex-b-PLA block copolymer is a promising new biomaterial for making controlled nanoparticles as drug delivery vehicles.      

Biodegradable poly(amine-co-ester) terpolymers for targeted gene delivery

Many synthetic polycationic vectors for non-viral gene delivery show high efficiency in vitro, but their usually excessive charge density makes them toxic for in vivo applications. Here we describe the synthesis of a series of high molecular weight terpolymers with low charge density, and show that they exhibit efficient gene delivery, some surpassing the efficiency of the commercial transfection reagents Polyethylenimine and Lipofectamine 2000. The terpolymers were synthesized via enzyme-catalyzed copolymerization of lactone with dialkyl diester and amino diol, and their hydrophobicity adjusted by varying the lactone content and by selecting a lactone comonomer of specific ring size. Targeted delivery of the pro-apoptotic TRAIL gene to tumour xenografts by one of the terpolymers results in significant inhibition of tumour growth, with minimal toxicity both in vitro and in vivo. Our findings suggest that the gene delivery ability of the terpolymers stems from their high molecular weight and increased hydrophobicity, which compensates for their low charge density.     

Application of rotatable central composite design in the preparation and optimization of poly(lactic-co-glycolic acid) nanoparticles for controlled delivery of paclitaxel

Context: Polymeric carrier systems of paclitaxel (PCT) offer advantages over only available formulation Taxol® in terms of enhancing therapeutic efficacy and eliminating adverse effects. Objective: The objective of the present study was to prepare poly (lactic-co-glycolic acid) nanoparticles containing PCT using emulsion solvent evaporation technique. Methods: Critical factors involved in the processing method were identified and optimized by scientific, efficient rotatable central composite design aiming at low mean particle size and high entrapment efficiency. Twenty different experiments were designed and each formulation was evaluated for mean particle size and entrapment efficiency. The optimized formulation was evaluated for in vitro drug release, and absorption characteristics were studied using in situ rat intestinal permeability study. Results: Amount of polymer and duration of ultrasonication were found to have significant effect on mean particle size and entrapment efficiency. First-order interactions of amount of miglyol with amount of polymer were significant in case of mean particle size, whereas second-order interactions of polymer were significant in mean particle size and entrapment efficiency. The developed quadratic model showed high correlation (R² > 0.85) between predicted response and studied factors. The optimized formulation had low mean particle size (231.68 nm) and high entrapment efficiency (95.18%) with 4.88% drug content. The optimized formulation showed controlled release of PCT for more than 72 hours. In situ absorption study showed faster and enhanced extent of absorption of PCT from nanoparticles compared to pure drug. Conclusion: The poly (lactic-co-glycolic acid) nanoparticles containing PCT may be of clinical importance in enhancing its oral bioavailability.     

Synthesis and cytocompatibility of manganese (II) and iron (III) substituted hydroxyapatite nanoparticles

Manganese (II) and iron (III) substituted hydroxyapatite (HA, Ca₁₀(PO₄)₆(OH)₂) nanoparticles were synthesized using wet chemical method. All samples were single-phase, non-stoichiometric and B-type carbonated hydroxyapatite. Compared with pure HA, Mn²⁺ substituted (MnHA) and Fe³⁺ doped HA (FeHA) did not demonstrate significant structure deviation. Since ion exchange mechanism was applied for the synthesis process, the morphology and particle size were not significantly affected: all samples were elongated spheroids of around 70 nm. The presence of Fe and Mn was confirmed by energy dispersive X-ray spectroscopy (EDX) while the concentrations were quantified by inductively coupled plasma (ICP). Fe³⁺ ions were more active than Mn²⁺ ions in replacing Ca²⁺ ions in HA lattice structure. The magnetic property of HA was modified by substitution with Fe. The Fe5 (Fe_added/Ca_added = 5% by molar ratio) was paramagnetic while pure HA was diamagnetic. Results of extraction assay from cells cultured in extracted medium for 72 h suggested that both MnHA and FeHA were non-cytotoxic to osteoblast cells. Meanwhile, the presence of Fe³⁺ ions in HA demonstrated significant positive effect on osteoblast cells, where the cell number on Fe5 pellets was twice that of pure HA and MnHA samples.     

Zinc-doped hydroxyapatite and poly(propylene fumarate) nanocomposite scaffold for bone tissue engineering

Hydroxyapatite (HA) is a bioceramic material that shares similar crystal and chemical structures with inorganic components of the bone. However, HA lacks osteoinductive activity and has a brittle nature, making it challenging to apply for direct load-bearing bone applications. In this study, we used a wet chemical method to synthesize zinc-doped hydroxyapatite powders with different Zn/(Zn+Ca) molar ratios of 0, 0.025, 0.05, and 0.1. The corresponding Zn-HA was designated as HA, Zn2.5-HA, Zn5-HA, and Zn10-HA. The Zn-HA powders at 30 wt% were used to fabricate poly(propylene fumarate) (PPF)-based nanocomposite scaffolds (HA/PPF, Zn2.5-HA/PPF, Zn5-HA/PPF, and Zn10-HA/PPF). The physical properties of obtained scaffolds were examined by scanning electron microscopy, energy-dispersive X-ray spectroscopy (EDS), transmission electron microscopy (TEM), and atomic force microscopy (AFM). Live/dead cell viability assay showed that these scaffolds were biocompatible and supported excellent adhesion of MC3T3-E1 preosteoblast cells. Additionally, the proliferation of cells was detected at 1, 4, and 7 days on these scaffolds. Alkaline phosphatase (ALP) activity measurement and alizarin red staining showed good osteogenic differentiation and matrix mineralization for MC3T3-E1 cells growing on these scaffolds. Taken together, the results here indicate that Zn5-HA/PPF nanocomposite scaffolds are promising scaffold material for bone tissue engineering.

Graphical abstract

     

Scaffolds for drug delivery,part I: electrospun porous poly(lactic acid) and poly(lactic acid)/poly(ethylene oxide) hybrid scaffolds

This is the first in a series of papers, focused on the development of a biodegradable, controlled, and potentially targeted drug delivery system. In this paper, we describe the production of highly porous biodegradable fibrous structures suitable for biomedical applications and as a matrix for drug delivery. Two structures are described below. The first structure is composed of electrospun poly(lactic acid) (PLA) fibers and is unique due to (1) the uniformity if its constitute fibers’ diameter, (2) consistent surface pore dimensions of each fiber, (3) the use of only a single solvent, (4) interior nano-size porosity throughout each individual fiber, and (5) the independency of surface pore dimensions on fiber diameter. The produced matrix will be further impregnated with cargo loaded nanoparticles—Red clover necrotic mosaic virus (RCNMV)—to achieve a controlled drug delivery system (described in Part III) for cancer treatments. Such a structure can also be used as tissue engineering scaffolds and filter media. The second electrospun structure has enhanced hydrophilicity compared to PLA matrix and is formed by blending poly(lactic acid)/poly(ethylene oxide) (PEO) polymers. The incorporation of PEO in the matrix introduces preferable sites for aqueous compounds to be attached to while retaining the overall structural integrity and porous morphology. It is hypothesized that the existence of alternative hydrophilic and hydrophobic segments in the structure may reduce post-implantation complications such as platelet adhesion.     

OX26 modified hyperbranched polyglycerol-conjugated poly(lactic-co-glycolic acid) nanoparticles: synthesis, characterization and evaluation of its brain delivery ability

A novel nanoparticles-based brain drug delivery system made of hyperbranched polyglycerol-conjugated poly(lactic-co-glycolic acid) which was surface functionalized with transferrin antibody (OX26) was prepared. Hyperbranched polyglycerol-conjugated poly(lactic-co-glycolic acid) was synthesized, characterized and applied to prepare nanoparticles by means of double emulsion solvent evaporation technique. Transmission electron micrograph and dynamic light scattering showed that nanoparticles had a round and regular shape with a mean diameter of 170 ± 20 nm. Surface chemical composition was detected by X-ray photoelectron spectroscopy. Endomorphins, as a model drug, was encapsulated in the nanoparticles. In vitro drug release study showed that endomorphins was released continuously for 72 h. Cellular uptake study showed that the uptake of nanoparticles by the brain microvascular endothelial cells was both time- and concentration-dependant. Further uptake inhibition study indicated that the uptake of nanoparticles was via a caveolae-mediated endocytic pathway. In vivo endomorphins brain delivery ability was evaluated based upon the rat model of chronic constriction injury of sciatic nerve. OX26 modified nanoparticles had achieved better analgesic effects, compared with other groups. Thus, OX26 modified hyperbranched polyglycerol-conjugated poly(lactic-co-glycolic acid) nanoparticles may be a promising brain drug delivery carrier.     

原位聚合羟基磷灰石/聚醚醚酮复合材料

通过原位聚合法制备HA/PEEK复合材料,利用X射线衍射仪、红外光谱仪、差示扫描量热仪、扫描电镜对PEEK、HA/PEEK复合材料进行表征。研究表明,基体PEEK的聚合过程受到一定的影响;HA颗粒在基体之中有着优异的分散性。将PEEK、HA/PEEK复合材料压模成型,通过拉伸试验和硬度测量检测材料的力学性能,结果表明,HA的加入对复合材料的力学性能产生较大的影响。     

Fabrication of poly(D,L‐lactic acid) nanoparticles as delivery system for sustained release of L‐theanine

L‐theanine is present in tea as a unique, free, non‐protein amino acid. Due to various beneficial effects on brain activity, it is widely used as a nutraceutical. After consumption, it is rapidly absorbed and metabolised followed by excretion through urine. Therefore, the authors developed an L‐theanine delivery system by encapsulating into polymeric nanoparticles to release it slowly and make it available for a longer period of time. Poly(D, L‐lactic acid) nanoparticle (PLANP) was fabricated by the double emulsion method and L‐theanine was encapsulated into it (PLANP‐T). Spherical nanoparticles with a hydrodynamic diameter of 247 and 278 nm and surface charge of −14.5 and −25.7 mV for PLANP and PLANP‐T, respectively, were fabricated. The Fourier transform infrared spectroscopic data indicated encapsulation of L‐theanine into PLANP. The PLANP showed high L‐theanine encapsulation capacity (71.65%) with a sustained release character. The maximum release (66.3%) of L‐theanine was recorded in pH 7.3 at 48 h. The release kinetics followed the Higuchi model and the release mechanism was determined as super case‐II transport (erosion). This slow release will make it available to the target tissue for a longer period of time (sustain release effect) and will also avoid immediate metabolism and clearance from the circulation.Inspec keywords: nanomedicine, pH, polymers, nanofabrication, emulsions, biomedical materials, drug delivery systems, nanoparticles, Fourier transform infrared spectraOther keywords: brain activity, L‐theanine delivery system, polymeric nanoparticles, double emulsion method, spherical nanoparticles, surface charge, L‐theanine encapsulation capacity, poly(D, L‐lactic acid) nanoparticles, nonprotein amino acid, urine, hydrodynamic diameter, Fourier transform infrared spectroscopy, time 48.0 hour, voltage ‐25.7 mV, voltage ‐14.5 mV, size 278.0 nm, size 247.0 nm, target tissue, Higuchi model, pH