Amorphous magnesium phosphate-graphene oxide nano particles laden 3D-printed chitosan scaffolds with enhanced osteogenic potential and antibacterial properties.

The utilization of 3D printing technology for the fabrication of graft substitutes in bone repair holds immense promise. However, meeting the requirements for printability, bioactivity, mechanical strength, and biological properties of 3D printed structures concurrently poses a significant challenge. In this study, we introduce a novel approach by incorporating amorphous magnesium phosphate-graphene oxide (AMP-GO) into a thermo-crosslinkable chitosan/ß glycerol phosphate (CS/GP) ink. We fabricated thermo-crosslinkable CS inks containing varying concentrations (10 %, 20 %, or 30 % weight) of AMP-GO. The 3D printed scaffolds incorporating 20 % AMP-GO exhibited significantly improved mechanical properties, with compressive strengths of 4.5 ± 0.06 MPa compared to 0.5 ± 0.03 MPa for CS printed scaffolds. Moreover, the CS/AMP-GO inks demonstrated enhanced antibacterial activity against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) bacteria, attributed to the release of magnesium cations and the performance of GO. Additionally, CS/20AMP-GO ink facilitated increased adhesion, viability, proliferation, and osteogenic differentiation of mesenchymal stem cells (MSCs), as evidenced by the upregulation of ALP, COL1, and Runx2 expression, which were elevated 9.8, 6.5, and >22 times, respectively, compared to pure CS scaffolds. Considering its exceptional in vivo osteogenic potential, we anticipate that the CS/20AMP-GO ink holds great potential for 3D printing of bone grafts.

Kappa-carrageenan based hybrid hydrogel for soft tissue engineering applications.

Biological materials such as cell-derived membrane vesicles have emerged as alternative sources for molecular delivery systems, owing to multicomponent features, the inherent functionalities and signaling networks, and easy-to-carry therapeutic agents with various properties. Herein, red blood cell membrane (RBCM) vesicle-laden methacrylate kappa-carrageenan (KaMA) composite hydrogel is introduced for soft tissue engineering. Results revealed that the characteristics of hybrid hydrogels were significantly modulated by changing the RBCM vesicle content. For instance, the incorporation of 20% (v/v) RBCM significantly enhanced compressive strength from 103 ± 26 kPa to 257 ± 18 kPa and improved toughness under the cyclic loading from 1.0 ± 0.4 kJ m-3to 4.0 ± 0.5 kJ m-3after the 5thcycle. RBCM vesicles were also used for the encapsulation of curcumin (CUR) as a hydrophobic drug molecule. Results showed a controlled release of CUR over three days of immersion in PBS solution. The RBCM vesicles laden KaMA hydrogels also supportedin vitrofibroblast cell growth and proliferation. In summary, this research sheds light on KaMA/RBCM hydrogels, that could reveal fine-tuned properties and hydrophobic drug release in a controlled manner.

Antibacterial amorphous magnesium phosphate/graphene oxide for accelerating bone regeneration.

Magnesium phosphates (MgP)s have attracted interest as an alternative biomaterial compared to the calcium phosphate (CaP)s compounds in the bone regeneration application in terms of their prominent biodegradability, lack of cytotoxicity, and ability of bone repair stimulation. Among them, amorphous magnesium phosphates (AMP)s indicated a higher rate of resorption, while preserving high osteoblasts viability and proliferation, which is comparable to their CaP peers. However, fast degradation of AMP leads to the initial fast release of Mg2+ ions and adverse effects on its excellent biological features. It seems that the addition of graphene oxide (GO) to magnesium phosphate can moderate its degradation rate. Hence, a novel in situ synthesized AMP powders containing 0.05, 0.25, 0.5, and 1 wt% of graphene oxide (AMP/GO) were developed to achieve a favorable degradation rate, desirable antibacterial properties against both Escherichia coli (E. coli), Staphylococcus aureus (S. aureus) accompanying with proper cell viability and proliferation. The incorporation of 0.5 wt% of graphene oxide into the AMP ceramic led to reduce the release of Mg2+ ions from 571.2 ± 12.9 mg/L to 372.8 ± 14.7 mg/L and P ions from 354.8 ± 11.9 mg/L to 245.3 ± 9.9 mg/L, at day 10 of immersion in PBS. Besides, AMP/0.5 GO bioceramics were capable of eradicating all bacterial colonies of both strains. On the other hand, MG63 cells viability went up from 143.46% ± 7.54 to 184.46% ± 11.54 on the 7th day of culture in the presence of 0.5 wt% of GO compared to pure AMP ceramic. Furthermore, alizarin red staining and alkaline phosphatase (ALP) activity demonstrated the ability of AMP/GO to maintain the osteogenic phenotype of MG63 cells during 7 days culture. Therefore, it can be concluded that well distributed and in situ synthesized AMP/0.5GO powders can be a promising biomaterial for bone tissue regeneration.

Inspiring biomimetic system based on red blood cell membrane vesicles for effective curcumin loading and release.

The aim of this study is to introduce an inspiring biomimetic system based on the red blood cell membrane (RBCM) vesicles for improved encapsulation efficiency and release of curcumin (Cur). Here, the role of the sonication time (0.5, 1.5, 3 and 5 min) on the properties of RBCM-CUR vesicles is investigated. It is determined that the hydrodynamic vesicle size, zeta potential, and release behavior are tunable by changing the sonication time. Noticeably, the average size of vesicles decreased from 163.0 ± 21 nm to 116.3 ± 16 nm by increasing the sonication time from 0.5 to 5 min. Moreover, the drug release value, after 24 h incubation, enhances from 57 to 99% with the expansion of sonication from 0.5 to 5 min. Additionally, the entrapment efficiency of Cur as a model drug is high in whole sonication time, owing to the amphiphilic nature of RBCM. Finally, the RBCM-CUR vesicles are not only cytocompatible, but also could support the attachment and proliferation of fibroblast cells in vitro. The RBCM based system for delivery of Cur could be a promising system for the wound healing applications.

Shear-thinning and self-healing nanohybrid alginate-graphene oxide hydrogel based on guest-host assembly.

The study aims to develop a novel nanohybrid shear-thinning hydrogel with fast gelation, and variable mechanical and biological properties. This nanohybrid hydrogel was developed via self-assembly guest-host interaction between ß-cyclodextrin modified alginate (host macromere, Alg-CD) and adamantine modified graphene oxide (guest macromere, Ad-GO) and subsequent ionic crosslinking process. We found that the rheological and mechanical properties of hydrogels were controlled via macromere concentration and the host: guest macromere ratio, due to the modulation of crosslinking density and network structure. Noticeably, 12%(1:2) dual-crosslinked hydrogel (2DC12) significantly improved the strength (1.3-folds) and toughness compared to 10%(1:4) dual-crosslinked hydrogel (4DC10). Furthermore, the hydrogel erosion and cytocompatibility relied on the designed parameters. Remarkably, 2DC12 showed less than 20% weight loss after 20 days of incubation in physiological solution and more than 200% cell survival after five days. In conclusion, the nanohybrid Alg-GO hydrogel could be used as an injectable hydrogel for soft tissue engineering applications.

Biomimetic Nylon 6-Baghdadite Nanocomposite Scaffold for Bone Tissue Engineering.

Nature creates soft and hard ingredients revealing outstanding properties by adjusting the ordered assembly of simple primarily components from the nano- to the macro-scale. To simulate the important features of native tissue architecture, wide researches are being performed to develop new biomimetic custom-made composite scaffolds for tissue engineering. Here, we introduced a three-dimensional (3D) biomimetic scaffold based on the cuttlefish bone (CB) as a sacrificial template for bone tissue engineering. By combination of nylon 6 (N6), various amounts of baghdadite (BG) nanopowder and sacrificial template CB, a novel nanocomposite scaffolds was successfully developed with hierarchical microstructure and open pores in the range size in long and minor axis of 153-253 µm and 39-70 µm, respectively, depending on the BG content. In addition, incorporation of BG improved the mechanical properties of the scaffolds. Noticeably, the compressive strength and compressive modulus enhanced from 0.47 ± 0.05 to 1.41 ± 0.25 MPa and from 3.16 ± 0.14 to 6.23 ± 0.3 MPa, respectively. Moreover, results demonstrated that the incorporation of BG nanoparticles in the N6 matrix significantly improved bioactivity in simulated body fluid and increased degradation rate of N6 scaffold. Additionally, 3D nanocomposite scaffolds disclosed meaningfully excellent cellular responses. It is envisioned that the provided N6-BG nanocomposite scaffold might be a promising candidate for bone tissue engineering applications.

Design, fabrication and structural optimization of tubular carbon/Kevlar®/PMMA/graphene nanoplate composite for bone fixation prosthesis.

The present work investigates the mechanical properties of tubular carbon/Kevlar® composite coated with poly(methyl methacrylate)/graphene nanoplates as used in the internal fixation of bones. Carbon fibers are good candidates for developing high-strength biomaterials and due to better stress transfer and electrical properties, they can enhance tissue formation. In order to improve carbon brittleness, ductile Kevlar® was added to the composite. The tubular carbon/Kevlar® composites have been prepared with tailorable braiding technology by changing the fiber pattern and angle in the composite structure and the number of composite layers. Fuzzy analyses are used for optimizing the tailorable parameters of 80 prepared samples and then mechanical properties of selected samples are discussed from the viewpoint of mechanical properties required for a bone fixation device. Experimental results showed that with optimizing braiding parameters the desired composite structure with mechanical properties close to bone properties could be produced. Results showed that carbon/Kevlar® braid's physical properties, fiber composite distribution and diameter uniformity resulted in matrix uniformity, which enhanced strength and modulus due to better ability for distributing stress on the composite. Finally, as graphene nanoplates demonstrated their potential properties to improve wound healing intended for bone replacement, so reinforcing the PMMA matrix with graphene nanoplates enhanced the composite quality, for use as an implant.

Nano­calcium phosphate bone cement based on Si-stabilized α-tricalcium phosphate with improved mechanical properties.

This study aimed to develop nano­calcium phosphate cement (nCPC) and evaluate the effect of nanosized precursors on mechanical, physical and handling properties (injectability and setting time) as well as conversion rate of nano-reactants into nano-hydroxyapatite (nHA). In this study, while alpha tricalcium phosphate (α-TCP, 98wt%) and HA (2wt%) were applied as the powder phase, 2.5wt% NaH2PO4 solution was used as liquid phase of cement. Before nano-CPC preparation, Si-stabilized α-TCP nanopowder with particle size of 10±3.6nm was firstly synthesized in a two-step process of sol-gel followed by mechanical alloying. Moreover, HA nanopowder with particle size of 32±3.6nm was synthesized using sol-gel process. Our results revealed that after 3days of immersion in ringer's solution, reactants almost completely converted to nHA. Moreover, the initial and final setting time of nano-CPC was obtained 6.3±2.1min and 14.3±4.0min, respectively. Furthermore, injectability of this formulation was reached 87.90±2.60%. In addition, our results confirmed that the compressive strength and modulus of nano-CPC enhanced with increasing immersion time in ringer's solution from 9.50±1.27MPa and 0.38±0.07GPa (at 1day) to 18.70±2.23MPa and 0.57±0.15GPa (at 5days), respectively. Finally, in order to evaluate cellular responses to nano-CPC, MG63 cells were cultured on it and cell morphology and cytotoxicity were evaluated. Results revealed that nano-CPC enhanced proliferation and spreading of osteoblast like cells compared to control (tissue culture plate) which could be due to both appropriate physical and chemical properties of nano-CPC which stimulate cell proliferation. Our findings suggest the formation of an injectable nano-CPC with appropriate mechanical, physical and degradation rate which can potentially utilized for filling bone defects.

pH sensitive dexamethasone encapsulated laponite nanoplatelets: Release mechanism and cytotoxicity.

The purpose of this study was to develop an efficient strategy to use laponite (LAP) nanoplates as a platform for the efficient release of anionic dexamethasone (DEX). Results revealed that DEX was encapsulated into the interlayer space of LAP nanodisks through an intercalation process with a high loading efficiency of 95.10±0.80%. X-Ray diffraction (XRD) patterns and Fourier transform infrared (FTIR) spectra of the hybrid LAP/DEX nanoplates (LD-NPs) indicated that DEX molecules could successfully adsorb into the LAP nanoplates depending on the pH value. Moreover, in vitro drug release study showed that the release of DEX from LD-NPs was pH-dependent, and DEX released at a faster rate at acidic pH (pH=5.4) than physiological one. Importantly, the results of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay confirmed that the released DEX from LD-NPs not only did not show cytotoxic effect but also improved the viability of MG63 cells compared to LAP-free samples (DEX enriched medium). Our work indicated that LAP nanoplates could be a promising candidate for release of anionic DEX in the controlled manner depending on the pH environment. Moreover, the merits of LD-NPs such as good cytocompatibility, excellent physiological stability and sustained pH-responsive release properties, make them a promising platform for the delivery of other therapeutic agents beyond DEX.