3D Printing Assisted Fabrication of Copper-Silver Mesoporous Bioactive Glass Nanoparticles Reinforced Sodium Alginate/Poly(vinyl alcohol) Based Composite Scaffolds: Designed for Skin Tissue Engineering.

Additive manufacturing (also known as 3D printing) is a promising method for producing patient-specific implants. In the present study, sodium alginate (Na-ALG)/poly(vinyl alcohol) (PVA) polymer blends of varying ratios (1:0, 3:1, 1:1, and 1:3) were used to produce tailored-designed skin scaffolds using a 3D bioprinter. Samples of skin scaffolds were printed at 20 layers with a layer height of 0.15 mm using a needle with an inner diameter of 330 µm while maintaining the extrusion speed, extrusion width, and fill density at 10 mm/s, 0.2 mm, and 85%, respectively. The Na-ALG/PVA blend with a 3:1 ratio showed the best printability due to its good viscosity and minimal nozzle leakage, allowing for the fabrication of skin scaffolds with high fidelity and the desired morphological characteristics. Then, copper-silver doped mesoporous bioactive glass nanoparticles (Cu-Ag MBGNs) were incorporated into the Na-ALG/PVA blend (which had already been prepared by using a Na-ALG:PVA ratio of 3:1) in order to obtain therapeutic (angiogenic and antibacterial) effects. The fabricated Na-ALG/PVA/Cu-Ag MBGNs biocomposite scaffolds with dimensions of 20 mm× 20 × 3 mm3 and pore size of 400 ± 60 µm exhibited a promising fidelity. The presence of chemical bonds attributed to Na-ALG, PVA, and Cu-Ag MBGNs and the uniform distribution of Na, C, and O elements within the microstructure of the scaffolds were confirmed by EDX, SEM, and FTIR analyses. The scaffolds were hydrophilic and exhibited proper swelling and degradation behavior for skin tissue engineering. According to the inhibition halo test, the scaffolds exhibited strong antibacterial activity against Staphylococcus aureus and Escherichia coli. The cytocompatibility to human-derived fibroblast cells was confirmed by the WST-8 assay and in vivo Chorioallantoic Membrane Assay. In addition, Na-ALG/PVA/Cu-Ag MBGNs showed angiogenic potential, exhibiting favorable wound healing properties.

Development and Characterization of Chitosan and Chondroitin Sulfate Based Hydrogels Enriched with Garlic Extract for Potential Wound Healing/Skin Regeneration Applications.

Hydrogels can provide instant relief to pain and facilitate the fast recovery of wounds. Currently, the incorporation of medicinal herbs/plants in polymer matrix is being investigated due to their anti-bacterial and wound healing properties. Herein, we investigated the novel combination of chitosan (CS) and chondroitin sulfate (CHI) to synthesize hydrogels through freeze gelation process and enriched it with garlic (Gar) by soaking the hydrogels in garlic juice for faster wound healing and resistance to microbial growth at the wound surface. The synthesized hydrogels were characterized via Fourier-transform infrared spectroscopy (FTIR), which confirmed the presence of relevant functional groups. The scanning electron microscopy (SEM) images exhibited the porous structure of the hydrogels, which is useful for the sustained release of Gar from the hydrogels. The synthesized hydrogels showed significant inhibition zones against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). Furthermore, cell culture studies confirmed the cyto-compatibility of the synthesized hydrogels. Thus, the novel hydrogels presented in this study can offer an antibacterial effect during wound healing and promote tissue regeneration.

Development and Characterization of Zein/Ag-Sr Doped Mesoporous Bioactive Glass Nanoparticles Coatings for Biomedical Applications.

Implants are used to replace damaged biological structures in human body. Although stainless steel (SS) is a well-known implant material, corrosion of SS implants leads to the release of toxic metallic ions, which produce harmful effects in human body. To prevent material degradation and its harmful repercussions, these implanted materials are subjected to biocompatible coatings. Polymeric coatings play a vital role in enhancing the mechanical and biological integrity of the implanted devices. Zein is a natural protein extracted from corn and is known to have good biocompatibility and biodegradability. In this study, zein/Ag-Sr doped mesoporous bioactive glass nanoparticles (Ag-Sr MBGNs) were deposited on SS substrates via electrophoretic deposition (EPD) at different parameters. Ag and Sr ions were added to impart antibacterial and osteogenic properties to the coatings, respectively. In order to examine the surface morphology of coatings, optical microscopy and scanning electron microscopy (SEM) were performed. To analyze mechanical strength, a pencil scratch test, bend test, and corrosion and wear tests were conducted on zein/Ag-Sr doped MBGN coatings. The results show good adhesion strength, wettability, corrosion, and wear resistance for zein/Ag-Sr doped MBGN coatings as compared to bare SS substrate. Thus, good mechanical and biological properties were observed for zein/Ag-Sr doped MBGN coatings. Results suggested these zein/Ag-Sr MBGNs coatings have great potential in bone regeneration applications.

Fabrication of Gelatin Methacrylate (GelMA) Scaffolds with Nano- and Micro-Topographical and Morphological Features.

The design of biomimetic biomaterials for cell culture has become a great tool to study and understand cell behavior, tissue degradation, and lesion. Topographical and morphological features play an important role in modulating cell behavior. In this study, a dual methodology was evaluated to generate novel gelatin methacrylate (GelMA)-based scaffolds with nano and micro topographical and morphological features. First, electrospinning parameters and crosslinking processes were optimized to obtain electrospun nanofibrous scaffolds. GelMA mats were characterized by SEM, FTIR, DSC, TGA, contact angle, and water uptake. Various nanofibrous GelMA mats with defect-free fibers and stability in aqueous media were obtained. Then, micropatterned molds produced by photolithography were used as collectors in the electrospinning process. Thus, biocompatible GelMA nanofibrous scaffolds with micro-patterns that mimic extracellular matrix were obtained successfully by combining two micro/nanofabrication techniques, electrospinning, and micromolding. Taking into account the cell viability results, the methodology used in this study could be considered a valuable tool to develop patterned GelMA based nanofibrous scaffolds for cell culture and tissue engineering.

Systematically Designed Periodic Electrophoretic Deposition for Decorating 3D Carbon-Based Scaffolds with Bioactive Nanoparticles.

The coating of porous scaffolds with nanoparticles is crucial in many applications, for example to generate scaffolds for catalysis or to make scaffolds bioactive. A standard and well-established method for coating surfaces with charged nanoparticles is electrophoresis, but when used on porous scaffolds, this method often leads to a blockage of the pores so that only the outermost layers of the scaffolds are coated. In this study, the electrophoretic coating process is monitored in situ and the kinetics of nanoparticle deposition are investigated. This concept can be extended to design a periodic electrophoretic deposition (PEPD) strategy, thus avoiding the typical blockage of surface pores. In the present work we demonstrate successful and homogeneous electrophoretic deposition of hydroxyapatite nanoparticles (HAn, diameter ≤200 nm) on a fibrous graphitic 3D structure (ultralightweight aerographite) using the PEPD strategy. The microfilaments of the resulting scaffold are covered with HAn both internally and on the surface. Furthermore, protein adsorption assays and cell proliferation assays were carried out and revealed that the HAn-decorated aerographite scaffolds are biocompatible. The HAn decoration of the scaffolds also significantly increases the alkaline phosphatase activity of osteoblast cells, showing that the scaffolds are able to promote their osteoblastic activity.

Synthesis and characterization of manganese containing mesoporous bioactive glass nanoparticles for biomedical applications.

Mesoporous bioactive glass (BG) nanoparticles based in the system: SiO2-P2O5-CaO-MnO were synthesized via a modified Stöber process at various concentrations of Mn (0-7 mol %). The synthesized manganese-doped BG nanoparticles were characterized in terms of morphology, composition, in vitro bioactivity and antibacterial activity. Scanning electron microscopy (SEM), transmission electron microscopy (TEM) and Brunauer-Emmett-Teller (BET) analysis confirmed that the particles had spherical morphology (mean particle size: 110 nm) with disordered mesoporous structure. Energy dispersive X-ray spectroscopy (EDX) confirmed the presence of Mn, Ca, Si and P in the synthesized Mn-doped BG particles. Moreover, X-ray diffraction (XRD) analysis showed that Mn has been incorporated in the amorphous silica network (bioactive glass). Moreover, it was found that manganese-doped BG particles form apatite crystals upon immersion in simulated body fluid (SBF). Inductively coupled plasma atomic emission spectroscopy (ICP-OES) measurements confirmed that Mn is released in a sustained manner, which provided antibacterial effect against Bacillus subtilis, Pseudomonas aeruginosa and Staphylococcus aureus. The results indicate that the incorporation of Mn in the bioactive glass network is an effective strategy to develop novel multifunctional BG nanoparticles for bone tissue engineering.