There exists a technological need for advanced materials with improved properties for emerging biomedical applications. Recent developments in macroporous materials have demonstrated their applicability as indispensable tools in biomedical research. Cryogels, which are materials with a macroporous 3D structure, are produced as a result of controlled freezing during polymerization with a highly interconnected polymer network. Cryogels’ interest lies in their ability to address some of the limitations of their hydrogel analogues. In this review, hydrogel and cryogel basic concepts are discussed as a short primer for readers unfamiliar with the cryogels literature. Next, a general overview of the methods for synthesis and characterization of cryogels is provided, highlighting key concepts relevant to cryogels and explaining their unique properties. Finally an in‐depth overview of specific technologies and fields where cryogels have been applied is given. It is argued that the latest advances in cryogel technologies are able to address challenges in bioseparation, tissue engineering, and other emerging bioengineering disciplines.

Electrospinning is a versatile technique used to create native tissue-like fibrous scaffolds. Recently, it has gained a large amount of attention for generation of bioactive dressing materials suitable for treatment of both chronic and acute wounds. In this Review, we focus on the latest advances made in the application of electrospun scaffolds for bioactive wound healing. We first provide a brief overview of the wound healing process and electrospinning approaches. We then discuss fabrication of scaffolds made from natural and synthetic polymers via electrospinning for effective wound treatment and management. Natural polymers used for wound healing included in our Review cover protein based polymers such as collagen, gelatin, and silk and polysaccharide based polymers such as chitosan, hyaluronic acid, and alginate. In addition, we discuss aliphatic polyesters, super hydrophilic polymers, and polyurethanes as some of the most commonly used synthetic polymers for wound healing and wound dressing applications. Next, we review multifunctional and “smart” scaffolds developed by electrospinning based approaches. We place an emphasis on how flexibility of the electrospinning process enables production of advanced scaffolds such as core–shell fibrous scaffolds, multilayer scaffolds, and surface modified scaffolds. Taken together, it is clear that electrospinning is an emerging technology that provides a unique opportunity for engineering more effective wound dressing, management, and care products.

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The intracellular delivery of biologically active protein represents an important emerging strategy for both fundamental and therapeutic applications. Here, we optimized in vitro delivery of two functional proteins, the β-galactosidase (β-gal) enzyme and the anti-cytokeratin8 (K8) antibody, using liposome-based formulation. The guanidinium-cholesterol cationic lipid bis (guanidinium)-tren-cholesterol (BGTC) (bis (guanidinium)-tren-cholesterol) combined to the colipid dioleoyl phosphatidylethanolamine (DOPE) (dioleoyl phosphatidylethanolamine) was shown to efficiently deliver the β-gal intracellularly without compromising its activity. The lipid/protein molar ratio, protein amount, and culture medium were demonstrated to be key parameters affecting delivery efficiency. The protein itself is an essential factor requiring selection of the appropriate cationic lipid as illustrated by low K8 binding activity of the anti-K8 antibody using guanidinium-based liposome. Optimization of various lipids led to the identification of the aminoglycoside lipid dioleyl succinyl paromomycin (DOSP) associated with the imidazole-based helper lipid MM27 as a potent delivery system for K8 antibody, achieving delivery in 67% of HeLa cells. Cryo-transmission electron microscopy showed that the structure of supramolecular assemblies BGTC:DOPE/β-gal and DOSP:MM27/K8 were different depending on liposome types and lipid/protein molar ratio. Finally, we observed that K8 treatment with DOSP:MM27/K8 rescues the cyclic adenosine monophosphate (cAMP)-dependent chloride efflux in F508del-CFTR expressing cells, providing a new tool for the study of channelopathies.

Polymeric scaffolds such as hydrogels can be engineered to restore, maintain, or improve impaired tissues and organs. However, most hydrogels require surgical implantation that can cause several complications such as infection and damage to adjacent tissues. Therefore, developing minimally invasive strategies is of critical importance for these purposes. Herein, we developed several injectable cryogels made out of hyaluronic acid and gelatin for tissue-engineering applications. The physicochemical properties of hyaluronic acid combined with the intrinsic cell-adhesion properties of gelatin can provide suitable physical support for the attachment, survival, and spreading of cells. The physical characteristics of pure gelatin cryogels, such as mechanics and injectability, were enhanced once copolymerized with hyaluronic acid. Reciprocally, the adhesion of 3T3 cells cultured in hyaluronic acid cryogels was enhanced when formulated with gelatin. Furthermore, cryogels had a minimal effect on bone marrow dendritic cell activation, suggesting their cytocompatibility. Finally, in vitro studies revealed that copolymerizing gelatin with hyaluronic acid did not significantly alter their respective intrinsic biological properties. These findings suggest that hyaluronic acid-co-gelatin cryogels combined the favorable inherent properties of each biopolymer, providing a mechanically robust, cell-responsive, macroporous, and injectable platform for tissue-engineering applications.

The intracellular delivery of nucleic acid molecules is a complex process involving several distinct steps; among these the endosomal escape appeared to be of particular importance for an efficient protein production (or inhibition) into host cells. In the present study, a new series of ionizable vectors, derived from naturally occurring aminoglycoside tobramycin, was prepared using improved synthetic procedures that allow structural variations on the linker and hydrophobic domain levels. Complexes formed between the new ionizable lipids and mRNA, DNA, or siRNA were characterized by cryo-TEM experiments and their transfection potency was evaluated using different cell types. We demonstrated that lead molecule 30, bearing a biodegradable diester linker, formed small complexes with nucleic acids and provided very high transfection efficiency with all nucleic acids and cell types tested. The obtained results suggested that the improved and "universal" delivery properties of 30 resulted from an optimized endosomal escape, through the lipid-mixing mechanism.

Prior to any clinical application, terminal sterilization of biomaterials is a critical process imposed by the Food and Drug Administration. Of all the methods available for sterilization, high‐pressure steam sterilization such as autoclaving is the most widely used. While autoclave sterilization minimizes pathogen contamination, it can dramatically impact both structural and biological properties of biomaterials. It has recently been reported that injectable cryogels with shape memory properties hold great promises as 3D macroporous biomimetic scaffolds for biomedical applications including tissue engineering. In this study, the impact of autoclave sterilization on properties of a series of cryogels is measured. Unlike conventional hydrogels, cryogels made of natural polymers demonstrate a strong resilience to autoclave sterilization. This process does not alter either their macrostructural or unique physical properties including syringe injectability. The scaffolds' bioactive sites are preserved and autoclaved cryogels retain their excellent cytological compatibility post‐autoclaving. Furthermore, autoclaved cryogels do not trigger a notable activation of primary murine bone marrow‐derived dendritic cells suggesting a minimal risk for biomaterial‐induced inflammation, which is further confirmed by an in vivo histologic analysis. In summary, these results further demonstrate the huge potential of cryogels in the biomedical field and their capacity to be translated into clinical applications.

For several biomedical applications, it is essential to develop novel bioactive materials. Such biomaterials could potentially improve wound healing, prevent infections, or be used in immunoengineering. For example, bioactive materials that reduce oxidative stress without relying on antibiotics and other drugs could be beneficial. Hydrogel-based biomaterials, especially those derived from natural polymers, have been regarded as one of the most promising scaffolds for biomedical research. These multifunctional scaffolds can exhibit high water adsorption capacity, biocompatibility, and biomechanical properties that can match native tissues. Cryogels are a special type of hydrogels in which polymers are cross-linked around ice crystals. As a result, cryogels exhibit unique physical features, including a macroporous and interconnected network, flexibility, shape-memory properties, and syringe injectability. Herein, we developed a multifunctional, i.e., antibacterial, antioxidant, and injectable cryogel by combining lignin with gelatin. The cryogel with 0.2% lignin showed a compressive modulus of 25 kPa and a compressive stress of 140 kPa at 80% strain, which is, respectively, 1.8 and 7 times higher than those of the pure gelatin cryogels. Meanwhile, such a cryogel formulation could completely recover its shape after compression up to 90% and was needle-injectable. Additionally, the lignin-co-gelatin cryogel with 0.1–0.2 lignin showed 8–10 mm of inhibition zone against the most common surgical site infection-associated pathogenic bacteria. Furthermore, lignin-co-gelatin cryogel was found to scavenge free radicals and have good cytocompatibility, and the cryogels with up to 0.2% lignin minimally activate naïve mouse bone marrow-derived dendritic cells. Overall, the current approach shows great promise for the design of bioresource-based multifunctional cryogels for a wide range of biomedical applications.