Protein by-products: Composition, extraction, and biomedical applications.

Significant upsurge in animal by-products such as skin, bones, wool, hides, feathers, and fats has become a global challenge and, if not properly disposed of, can spread contamination and viral diseases. Animal by-products are rich in proteins, which can be used as nutritional, pharmacologically functional ingredients, and biomedical materials. Therefore, recycling these abundant and renewable by-products and extracting high value-added components from them is a sustainable approach to reclaim animal by-products while addressing scarce landfill resources. This article appraises the most recent studies conducted in the last five years on animal-derived proteins' separation and biomedical application. The effort encompasses an introduction about the composition, an overview of the extraction and purification methods, and the broad range of biomedical applications of these ensuing proteins.

Bioinspired Processing: Complex Coacervates as Versatile Inks for 3D Bioprinting.

3D bioprinting is a powerful fabrication technique in biomedical engineering, which is currently limited by the number of available materials that meet all physicochemical and cytocompatibility requirements for biomaterial inks. Inspired by the key role of coacervation in the extrusion and spinning of many natural materials, hyaluronic acid-chitosan complex coacervates are proposed here as tunable biomaterial inks. Complex coacervates are obtained through an associative liquid-liquid phase separation driven by electrostatic attraction between oppositely charged macromolecules. They offer bioactive properties and facile modulation of their mechanical properties through mild physicochemical changes in the environment, making them attractive for 3D bioprinting. Fine-tuning the salt concentration, pH, and molecular weight of the constituent polymers results in biomaterial inks that are printable in air and water. The biomaterial ink, initially a viscoelastic fluid, transitions into a viscoelastic solid upon printing due to dehydration (for printing in air) or due to a change in pH and ionic composition (for printing in solution). Consequently, scaffolds printed using the complex coacervate inks are stable without the need for post-printing processing. Fabricated cell culture scaffolds are cytocompatible and show long-term topological stability. These results pave the way to a new class of easy-to-handle tunable biomaterials for biofabrication.

Development of Ag NPs/allantoin loaded PCL/GEL electrospun nanofibers for topical wound treatment.

In the present study, the allantoin and silver nanoparticle (Ag NPs) loaded poly caprolactone/gelatin (PCL/GEL) nanofibers produced using electrospinning technique and their cyto-compatibility and wound healing activity were evaluated in vitro and in vivo. The SEM imaging revealed diameters of 278.8 ± 10 and 240.6 ± 12 nm for PCL/GEL/Ag NPs and PCL/GEL/Ag NPs/allantoin scaffolds. The Ag NPs entrapment into scaffolds was evaluated by FTIR analysis and EDX mapping. Both scaffolds containing Ag NPs and Ag NPs/allantoin exhibited valuable wound healing activity in Wistar rat animal model. The profound granulation tissue formation, high collagen deposition in coordination with low level of edema and inflammatory cells in Ag NPs/allantoin loaded scaffolds resulted in complete and mature re-epithelialization in giving the healing score (12 out of 12) equal to positive control group to the wounds treated with these scaffolds. It was concluded that the Ag NPs/allantoin loaded scaffolds regarding to their good antibacterial activity and excellent wound healing activity could be introduced as new effective wound dressing materials.

Effect of Dynamically Arrested Domains on the Phase Behavior, Linear Viscoelasticity and Microstructure of Hyaluronic Acid - Chitosan Complex Coacervates.

Complex coacervates make up a class of versatile materials formed as a result of the electrostatic associations between oppositely charged polyelectrolytes. It is well-known that the viscoelastic properties of these materials can be easily altered with the ionic strength of the medium, resulting in a range of materials from free-flowing liquids to gel-like solids. However, in addition to electrostatics, several other noncovalent interactions could influence the formation of the coacervate phase depending on the chemical nature of the polymers involved. Here, the importance of intermolecular hydrogen bonds on the phase behavior, microstructure, and viscoelasticity of hyaluronic acid (HA)-chitosan (CHI) complex coacervates is revealed. The density of intermolecular hydrogen bonds between CHI units increases with increasing pH of coacervation, which results in dynamically arrested regions within the complex coacervate, leading to elastic gel-like behavior. This pH-dependent behavior may be very relevant for the controlled solidification of complex coacervates and thus for polyelectrolyte material design.

Preparation of multilayer electrospun nanofibrous scaffolds containing soluble eggshell membrane as potential dermal substitute.

Electrospinning of natural and synthetic polymers has shown to be a great candidate for the fabrication of tissue engineering scaffolds due to their similarity to the nanofibrous structure of natural extracellular matrix (ECM). Moreover, the addition of ECM-like proteins could enhance the biocompatibility of these scaffolds. In this study, soluble eggshell protein (SEP) was first extracted and synthesized from the raw eggshell membrane. The characteristics and biocompatibility of the extracted SEP were evaluated using attenuated total reflectance-Fourier transform infrared (ATR-FTIR) analysis and 3-(4,5- dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide) (MTT) assay. For scaffolds fabrication, a three-layer nanofibrous composite structure was produced using the electrospinning technique. The outer layers composed of polyvinyl alcohol, chitosan, and extracted SEP while the middle layer composed of polyethylene oxide, gelatin, and zinc oxide nanoparticles (ZnO-NPs). For each layer, the electrospinning parameters were adjusted to form bead-free fibers. To improve fibers' stability against body fluids, the produced fibers were crosslinked using glutaraldehyde vapor. Several techniques such as scanning electron microscopy (SEM), energy dispersive X-ray, ATR-FTIR, swelling, tensile test, in vitro biodegradation, and MTT assay were implemented to evaluate the physical, chemical, and biological characterization of the fabricated fibers. The results showed that crosslinked fibers have adequate stability in water, suitable mechanical properties, and promising water uptake capacity. The MTT results also revealed that SEP and ZnO-NPs could increase scaffolds biocompatibility. Moreover, SEM photographs of cultured fibroblasts cells on the scaffolds showed that cells were well attached on the scaffolds and preserve their natural spindle shapes. Altogether, our findings demonstrated that the produced three-layer composite scaffolds are potential candidates for skin tissue engineering.

Avian Egg: A Multifaceted Biomaterial for Tissue Engineering.

Most components in avian eggs, offering a natural and environmentally friendly source of raw materials, hold great potential in tissue engineering. An avian egg consists of several beneficial elements: the protective eggshell, the eggshell membrane, the egg white (albumen), and the egg yolk (vitellus). The eggshell is mostly composed of calcium carbonate and has intrinsic biological properties that stimulate bone repair. It is a suitable precursor for the synthesis of hydroxyapatite and calcium phosphate, which are particularly relevant for bone tissue engineering. The eggshell membrane is a thin protein-based layer with a fibrous structure and is constituted of several valuable biopolymers, such as collagen and hyaluronic acid, that are also found in the human extracellular matrix. As a result, the eggshell membrane has found several applications in skin tissue repair and regeneration. The egg white is a protein-rich material that is under investigation for the design of functional protein-based hydrogel scaffolds. The egg yolk, mostly composed of lipids but also diverse essential nutrients (e.g., proteins, minerals, vitamins), has potential applications in wound healing and bone tissue engineering. This review summarizes the advantages and status of each egg component in tissue engineering and regenerative medicine, but also covers their current limitations and future perspectives.

Bioactive antibacterial bilayer PCL/gelatin nanofibrous scaffold promotes full-thickness wound healing.

Treatment of diabetic, chronic, and full-thickness wounds is a challenge as these injuries usually lead to infections that cause delayed and inappropriate healing. Therefore, fabrication of skin scaffolds with prolonged antibacterial properties are of great interest. Due to this demand, bilayered nanofibrous scaffolds were fabricated based on polycaprolactone and gelatin. The top layer of these scaffolds contained amoxicillin as a model drug and the bottom layer was loaded with zinc oxide nanoparticles to accelerate wound healing. Several characterization techniques including FTIR, SEM, swelling, tensile test, in vitro degradation, drug release, antibacterial activity, and MTT assay were used to assess physical, mechanical, and biological properties of produced nanofibers. SEM results demonstrated that bilayered scaffolds have smooth bead-free microstructures while in vitro release test showed that samples have a sustained release for amoxicillin up to 144 h (tested time). Disk diffusion assessment confirmed the potency of scaffolds for hindering bacterial growth while results of cytotoxicity evaluation revealed that scaffolds could effectively accelerate cell proliferation. Finally, in vivo tests on full-thickness rat models revealed that fabricated nanofibers accelerate wound contraction, increase collagen deposition and angiogenesis, and prevent scar formation. Altogether, results showed that fabricated scaffolds are promising candidates for treatment of full-thickness wounds.

Vascularization strategies for skin tissue engineering.

A number of challenges in skin grafting for wound healing have drawn researchers to focus on skin tissue engineering as an alternative solution. The core idea of tissue engineering is to use scaffolds, cells, and/or bioactive molecules to help the skin to properly recover from injuries. Over the past decades, the field has significantly evolved, developing various strategies to accelerate and improve skin regeneration. However, there are still several concerns that should be addressed. Among these limitations, vascularization is known as a critical challenge that needs thorough consideration. Delayed wound healing of large defects results in an insufficient vascular network and ultimately ischemia. Recent advances in the field of tissue engineering paved the way to improve vascularization of skin substitutes. Broadly, these solutions can be classified into two categories as (1) use of growth factors, reactive oxygen species-inducing nanoparticles, and stem cells to promote angiogenesis, and (2) in vitro or in vivo prevascularization of skin grafts. This review summarizes the state-of-the-art approaches, their limitations, and highlights the latest advances in therapeutic vascularization strategies for skin tissue engineering.