Enhanced osteogenic and bactericidal performance of premixed calcium phosphate cement with photocrosslinked alginate thin film.

The increasing prevalence of implant-associated infections (IAI) in orthopedics remains a public health challenge. Calcium phosphates (CaPs) are critical biomaterials in dental treatments and bone regeneration. It is highly desirable to endow CaPs with antibacterial properties. To achieve this purpose, we developed a photocrosslinked methacrylated alginate co-calcium phosphate cement (PMA-co-PCPC) with antibacterial properties, using α-tricalcium phosphate (α-TCP) powders with 16% amorphous contents as solid phase, liquid phases containing CuCl2 and SrCl2 as an inhibitor, and CaCl2 as an activator to construct PCPC. When CaCl2 started to activate the hydration reaction, Sr2+ or Cu2+ ions were exchanged with Ca2+, and α-TCP dissolution was restarted and gradually hydrated to form calcium-deficient hydroxyapatite (CDHA). PMA was added to crosslink with Cu/Sr ions and form gel-layer-wrapped hydrated CDHA. This study explored the binding mechanism of PMA and PCPC and the ion release rule of Ca2+ → Sr2+/Cu2+, optimized the construction of several antibacterial PMA-co-PCPC materials, and analyzed the physical, chemical, and biological properties. Because of the combined effect of Cu and Sr ions, the scaffold exhibited a potential antibacterial activity, promoting bone formation and vascular regeneration. This work provides a basis for designing antibacterial calcium phosphate biomaterials with controllable treatment, which is an important characteristic for preventing IAI of biomaterials.

Macroporous zwitterionic composite cryogel based on chitosan oligosaccharide for antifungal application.

Chitosan oligosaccharide (COS), a time-dependent antimicrobial carbohydrate, is found antifungal active with a short duration of action due to excessive solubility. We attempted to address this issue by employing a hydrogel as a COS carrier. In this research, macroporous zwitterionic composite cryogels composed of COS and poly(N-methacryl arginine) (PMarg) were fabricated, serving as long-term antifungal dressings. Firstly, Marg was synthesized and characterized by Fourier transform infrared spectroscopy (FT-IR), 1H and 13C nuclear magnetic resonance (NMR), and high-resolution mass spectrometry (HRMS). Then, the COS/PMarg cryogels were prepared by redox initiation cryopolymerization. The macroporous morphology of the cryogels was confirmed by scanning electron microscope (SEM) with pore size varying from 20.86 to 50.87 µm. FTIR indicated that hydrogen bonding formed between COS and PMarg, and the interaction elevated thermal stability of the cryogels as evidenced by thermal-gravimetric analysis (TGA). Swelling capacity, mechanical properties, and COS release behavior of the COS/PMarg cryogels were investigated. With the release of COS, the antifouling activity of the cryogel increased. Antimicrobial tests indicated the COS/PMarg cryogel could effectively inhibit the proliferation of Candida albicans. It demonstrated that the macroporous zwitterionic COS/PMarg composite cryogel might be a potential antifungal dressing with sequential "sterilization-release" capacity.

A series of carboxymethyl cellulose-based antimicrobial peptide mimics were synthesized for antimicrobial applications.

Inspired by antimicrobial peptides (AMP) which could alleviate drug resistance pressure, antimicrobial peptide mimics (AMPMs) were designed timely. Here, carboxymethyl cellulose (CMC) -based AMPMs were constructed by introducing different diamines on CMC effectively. Firstly, CMC was degraded to be oligomers with different molecular weights, followed by amination reactions with different diamines respectively. After protonation, a series of AMPMs with different structures were synthesized successfully. Their antibacterial effect has been evaluated by dynamic growth curves and microdilution method. The images snapped by the confocal laser scanning microscope and transmission electron microscope have fully proved its great lethality. And the antibacterial mechanism measured by flow cytometry analysis and zeta potential detection demonstrated that the destruction of membrane potential leads to bacteria death. The excellent blood compatibility and negligible drug resistance has also been confirmed. In addition, the synthesis method is simple and environmental-friendly.

Nanocopper-loaded Black phosphorus nanocomposites for efficient synergistic antibacterial application.

Novel nanocopper-loaded black phosphorus (BP/Cu) nanocomposites were synthesized to synergistically exert enhanced antibacterial activities aimed at reducing antibiotics abuse. First, both BP and Cu display low biotoxicity, broadening their application in the microbiological field. Second, the unique electronic properties of BP enable BP/Cu nanocomposites to amplify antibacterial effects via interfacial charge transfer, resulting in a surge of reactive oxygen species (ROS). Third, BP/Cu nanocomposites are relatively stable, which helps to avoid the problem that nanocopper alone is highly oxidized. Finally, BP/Cu was synthesized in an environmentally-friendly manner by a one-step reduction method. The BP/Cu nanocomposites were characterized by transmission electron microscopy and atomic force microscopy. Their antibacterial properties were investigated comprehensively and discussed in detail by inhibition zone assays, dynamic growth curves, membrane potential assays, and live/dead baclight bacterial viability assays, all of which revealed the antimicrobial activities of BP/Cu nanocomposites. Absorption spectra were measured to determine which ROS species were responsible for the bactericidal mechanisms. In summary, our results demonstrated the potential of nanocomposites based on BP in antibacterial therapy due to its excellent electronic properties and outstanding biological performance. This will pave the way for avoiding antibiotic overuse and for providing security to humans and the environment.

Osteogenic, Angiogenic, and Antibacterial Bioactive Nano-Hydroxyapatite Co-Synthesized Using γ-Polyglutamic Acid and Copper.

Nano-antibacterial calcium phosphate (CaP) has attracted intense attention with regard to its wide variety of medical and biological applications. The γ-polyglutamic acid and copper cosynthesized hydroxyapatite (γ-PGA/CuxHAp) was synthesized using the wet method. Structural and chemical characterizations demonstrate that copper was quantitatively incorporated into the hydroxyapatite structure, and the degree of Cu substitution was up to 20 mol % in the synthesized nanocrystals. Morphology characterization showed that the size of the γ-PGA/CuxHAp nanoparticles decreases with the increased copper content. γ-PGA/CuxHAp exhibited a steady release of Cu ions. Two experimental protocols were applied to compare the antibacterial activity of the γ-PGA/CuxHAp samples. A positive correlation was observed between Cu content and the inhibition of bacterial growth. The study also showed that nanoparticles with smaller particle sizes exhibited higher antibacterial activities than the larger particles. Endothelial and osteoblast cells rapidly proliferated on γ-PGA/CuxHAp, whereas high concentrations (20 mol %) of Cu ions reduced cell proliferation. In the rat calvarial defect model, some γ-PGA/CuxHAp samples such as γ-PGA/CuxHAp (x = 8, 16) showed efficient bone regeneration capacities at 12 weeks post implantation. Thus, the multibiofunctional γ-PGA/CuxHAp nanocomposite exhibited degradative, angiogenic, bactericidal and bone regenerative properties, providing a potential means to address some of the critical challenges in the field of bone tissue engineering.

Complexation and conformation of lead ion with poly-γ-glutamic acid in soluble state.

Complexation of microbial polymer in soluble state could impact the solubility, mobility, and bioavailability of heavy metals in the environment. The complexation of a bacterial exopolymer, poly-γ-glutamic acid (γ-PGA), with Pb2+ was studied using the polarographic method and circular dichroism measurement in soluble state. The number of available binding sites was determined based on the Chau's method and was found to be 0.04, 1.12, 3.56 and 4.51 mmol/(g dry weight of γ-PGA) at pH 3.4, 4.2, 5.0 and 6.2, respectively. Further, the number of binding sites was determined based on the Ruzic's method and was found to be 3.60 and 4.41 mmol/(g dry weight of γ-PGA) for pH 5.0 and 6.2, respectively. The constant (expressed as log K) values were 5.8 and 6.0 at pH 5.0 and 6.2. Compared to biopolymers secreted by other microorganisms, such as extracellular polymeric substances extraction from activated sludge, γ-PGA was a more efficient Pb2+ carrier from pH 5.0 to 6.2. The secondary structure of γ-PGA varied significantly when Pb2+ added. Ca2+ or Mg2+ replace a portion of the adsorbed Pb2+. However, the portion of Pb2+ involved in changing the γ-PGA conformation was not easily replaced by Ca2+ and Mg2+.

Pyrolysis preparation of poly-γ-glutamic acid derived amorphous carbon nitride for supporting Ag and γ-Fe2O3 nanocomposites with catalytic and antibacterial activity.

Nanocomposites composed of Ag and γ-Fe2O3 nanoparticles within poly-γ-glutamic acid (γ-PGA) derived amorphous carbon nitride (a-CN) films (Ag + γ-Fe2O3@a-CN) were synthesized by a one-step facile pyrolysis strategy. Transmission electron microscopy analysis shows that Ag and γ-Fe2O3 nanoparticles were obtained in situ and homogeneously dispersed on the a-CN matrix. The average size of nanoparticles was 8.1 nm. The presence of γ-Fe2O3, Ag, and a-CN in the nanocomposite was confirmed by X-ray diffraction analysis, UV-visible absorption spectroscopy, Raman spectroscopy, and X-ray photoelectron spectroscopy. Ag + γ-Fe2O3@a-CN catalyzed the degradation of methyl orange. This catalyst was also recycled ten times by magnetic separation without any loss in efficiency. Ag + γ-Fe2O3@a-CN exhibited strong antibacterial activity against E. coli and S. aureus. It also exhibited excellent antibacterial activity even after 20 times magnetic recycling. This indicates that this is a promising recyclable antibacterial and catalyst for environmental applications.

Hydrogen bonding impact on chitosan plasticization.

The use of chitosan natural polymers to replace synthetic polymers is of interest as part of the green materials movement. However, a major challenge of using chitosan material is its rigid and brittle nature associated primarily with its intra- and inter-molecular hydrogen bonds creating a hydrogen bond network. Plasticizers are able to make chitosan flexible, hypothetically by destroying its hydrogen bond networks. Herein, we showed the importance and complicated nature of the chitosan's hydrogen bond network with respect to its flexibility, through a comparative study of glycerol and ionic liquids plasticizers. The results demonstrated that glycerol's hydrogen bonding was able to disrupt the chitosan's hydrogen bond network resulting in a flexible film, but ionic liquids, even with their very strong hydrogen bonding, were not able to plasticize chitosan. This result suggested that the plasticization phenomenon was more complicated than hydrogen bond disruption. A molecular level study by quantum chemistry calculation showed that the efficiency of glycerol as chitosan plasticizer was due to its single hydrogen bonding site, which breaks down the chitosan hydrogen bonding networks, and leave hydrophobic C-H ending groups to limit the formation of inter molecular hydrogen bonds.

Combined delivery of bone morphogenetic protein-2 and insulin-like growth factor-1 from nano-poly (γ-glutamic acid)/ß-tricalcium phosphate-based calcium phosphate cement and its effect on bone regeneration in vitro.

In this study, nano-doped calcium phosphate cement delivery systems (poly (γ-glutamic acid)/ß-tricalcium phosphate/calcium phosphate ceramics and nano (γ-glutamic acid)/ß-tricalcium phosphate/calcium phosphate ceramic) were fabricated, and low doses (10 µg/g) of two growth factors, insulin-like growth factor-1 and bone morphogenetic protein-2, were encapsulated then sequentially released. We characterized the delivery systems using Fourier transform infrared spectroscopy and X-ray diffraction and measured washout resistance and compressive strength, and thus optimized the most appropriate proportioning of delivery systems for the two growth factors. One of the growth factors was absorbed by the nano-poly (γ-glutamic acid)/ß-tricalcium phosphate, which was then mixed into the calcium phosphate ceramic solid phase to create a new solid phase calcium phosphate ceramic. Nano-poly (γ-glutamic acid)/ß-tricalcium phosphate/calcium phosphate ceramic carriers were then prepared by blending the new calcium phosphate ceramic solid phase powder with a solution of the remaining growth factor. The effects of different release patterns (studying sequential behavior) of insulin-like growth factor-1 and bone morphogenetic protein-2 on osteogenic proliferation and differentiation of the MC3t3-E1 mouse osteoblast cell were investigated. This combinational delivery system provided a controlled release of the two growth factors, in which nano-doping significantly affected their release kinetics. The incorporation of dual growth factors could potentially stimulate bone healing and promoting bone ingrowth processes at a low dose.

Poly (γ-glutamic acid)/beta-TCP nanocomposites via in situ copolymerization: Preparation and characterization.

A series biodegradable poly (γ-glutamic acid)/beta-tricalcium phosphate (γ-PGA/TCP) nanocomposites were prepared which were composed of poly-γ-glutamic acid polymerized in situ with ß-tricalcium phosphate and physiochemically characterized as bone graft substitutes. The particle size via dynamic light scattering, the direct morphological characterization via transmission electron microscopy and field emission scanning electron microscope, which showed that γ-PGA and ß-TCP were combined compactly at 80℃, and the γ-PGA/TCP nanocomposites had homogenous and nano-sized grains with narrow particle size distributions. The water uptake and retention abilities, in vitro degradation properties, cytotoxicity in the simulated medium, and protein release of these novel γ-PGA/TCP composites were investigated. Cell proliferation in composites was nearly twice than ß-TCP when checked in vitro using MC3T3 cell line. We also envision the potential use of γ-PGA/TCP systems in bone growth factor or orthopedic drug delivery applications in future bone tissue engineering applications. These observations suggest that the γ-PGA/TCP are novel nanocomposites with great potential for application in the field of bone tissue engineering.