Self-Lubricating, Living Contact Lenses.

The increasing prevalence of dry eye syndrome in aging and digital societies compromises long-term contact lens (CL) wear and forces users to regular eye drop instillation to alleviate discomfort. Here a novel approach with the potential to improve and extend the lubrication properties of CLs is presented. This is achieved by embedding lubricant-secreting biofactories within the CL material. The self-replenishable reservoirs autonomously produce and release hyaluronic acid (HA), a natural lubrication and wetting agent, long term. The hydrogel matrix regulates the growth of the biofactories and the HA production, and allows the diffusion of nutrients and HA for at least 3 weeks. The continuous release of HA sustainably reduces the friction coefficient of the CL surface. A self-lubricating CL prototype is presented, where the functional biofactories are contained in a functional ring at the lens periphery, outside of the vision area. The device is cytocompatible and fulfils physicochemical requirements of commercial CLs. The fabrication process is compatible with current manufacturing processes of CLs for vision correction. It is envisioned that the durable-by-design approach in living CL could enable long-term wear comfort for CL users and minimize the need for lubricating eye drops.

Contribution of the ELRs to the development of advanced in vitro models.

Developing in vitro models that accurately mimic the microenvironment of biological structures or processes holds substantial promise for gaining insights into specific biological functions. In the field of tissue engineering and regenerative medicine, in vitro models able to capture the precise structural, topographical, and functional complexity of living tissues, prove to be valuable tools for comprehending disease mechanisms, assessing drug responses, and serving as alternatives or complements to animal testing. The choice of the right biomaterial and fabrication technique for the development of these in vitro models plays an important role in their functionality. In this sense, elastin-like recombinamers (ELRs) have emerged as an important tool for the fabrication of in vitro models overcoming the challenges encountered in natural and synthetic materials due to their intrinsic properties, such as phase transition behavior, tunable biological properties, viscoelasticity, and easy processability. In this review article, we will delve into the use of ELRs for molecular models of intrinsically disordered proteins (IDPs), as well as for the development of in vitro 3D models for regenerative medicine. The easy processability of the ELRs and their rational design has allowed their use for the development of spheroids and organoids, or bioinks for 3D bioprinting. Thus, incorporating ELRs into the toolkit of biomaterials used for the fabrication of in vitro models, represents a transformative step forward in improving the accuracy, efficiency, and functionality of these models, and opening up a wide range of possibilities in combination with advanced biofabrication techniques that remains to be explored.

Development of bioactive catechol functionalized nanoparticles applicable for 3D bioprinting.

Efficient wound treatments to target specific events in the healing process of chronic wounds constitute a significant aim in regenerative medicine. In this sense, nanomedicine can offer new opportunities to improve the effectiveness of existing wound therapies. The aim of this study was to develop catechol bearing polymeric nanoparticles (NPs) and to evaluate their potential in the field of wound healing. Thus, NPs wound healing promoting activities, potential for drug encapsulation and controlled release, and further incorporation in a hydrogel bioink formulation to fabricate cell-laden 3D scaffolds are studied. NPs with 2 and 29 M % catechol contents (named NP2 and NP29) were obtained by nanoprecipitation and presented hydrodynamic diameters of 100 and 75 nm respectively. These nanocarriers encapsulated the hydrophobic compound coumarin-6 with 70% encapsulation efficiency values. In cell culture studies, the NPs had a protective effect in RAW 264.7 macrophages against oxidative stress damage induced by radical oxygen species (ROS). They also presented a regulatory effect on the inflammatory response of stimulated macrophages and promoted upregulation of the vascular endothelial growth factor (VEGF) in fibroblasts and endothelial cells. In particular, NP29 were used in a hydrogel bioink formulation using carboxymethyl chitosan and hyaluronic acid as polymeric matrices. Using a reactive mixing bioprinting approach, NP-loaded hydrogel scaffolds with good structural integrity, shape fidelity and homogeneous NPs dispersion, were obtained. The in vitro catechol NPs release profile of the printed scaffolds revealed a sustained delivery. The bioprinted scaffolds supported viability and proliferation of encapsulated L929 fibroblasts over 14 days. We envision that the catechol functionalized NPs and resulting bioactive bioink presented in this work offer promising advantages for wound healing applications, as they: 1) support controlled release of bioactive catechol NPs to the wound site; 2) can incorporate additional therapeutic functions by co-encapsulating drugs; 3) can be printed into 3D scaffolds with tailored geometries based on patient requirements.

Emerging Biofabrication Techniques: A Review on Natural Polymers for Biomedical Applications.

Natural polymers have been widely used for biomedical applications in recent decades. They offer the advantages of resembling the extracellular matrix of native tissues and retaining biochemical cues and properties necessary to enhance their biocompatibility, so they usually improve the cellular attachment and behavior and avoid immunological reactions. Moreover, they offer a rapid degradability through natural enzymatic or chemical processes. However, natural polymers present poor mechanical strength, which frequently makes the manipulation processes difficult. Recent advances in biofabrication, 3D printing, microfluidics, and cell-electrospinning allow the manufacturing of complex natural polymer matrixes with biophysical and structural properties similar to those of the extracellular matrix. In addition, these techniques offer the possibility of incorporating different cell lines into the fabrication process, a revolutionary strategy broadly explored in recent years to produce cell-laden scaffolds that can better mimic the properties of functional tissues. In this review, the use of 3D printing, microfluidics, and electrospinning approaches has been extensively investigated for the biofabrication of naturally derived polymer scaffolds with encapsulated cells intended for biomedical applications (e.g., cell therapies, bone and dental grafts, cardiovascular or musculoskeletal tissue regeneration, and wound healing).

3D Printing of a Reactive Hydrogel Bio-Ink Using a Static Mixing Tool.

Hydrogel-based bio-inks have recently attracted more attention for 3D printing applications in tissue engineering due to their remarkable intrinsic properties, such as a cell supporting environment. However, their usually weak mechanical properties lead to poor printability and low stability of the obtained structures. To obtain good shape fidelity, current approaches based on extrusion printing use high viscosity solutions, which can compromise cell viability. This paper presents a novel bio-printing methodology based on a dual-syringe system with a static mixing tool that allows in situ crosslinking of a two-component hydrogel-based ink in the presence of living cells. The reactive hydrogel system consists of carboxymethyl chitosan (CMCh) and partially oxidized hyaluronic acid (HAox) that undergo fast self-covalent crosslinking via Schiff base formation. This new approach allows us to use low viscosity solutions since in situ gelation provides the appropriate structural integrity to maintain the printed shape. The proposed bio-ink formulation was optimized to match crosslinking kinetics with the printing process and multi-layered 3D bio-printed scaffolds were successfully obtained. Printed scaffolds showed moderate swelling, good biocompatibility with embedded cells, and were mechanically stable after 14 days of the cell culture. We envision that this straightforward, powerful, and generalizable printing approach can be used for a wide range of materials, growth factors, or cell types, to be employed for soft tissue regeneration.

Bioadhesive functional hydrogels: Controlled release of catechol species with antioxidant and antiinflammatory behavior.

Chronic wounds are particularly difficult to heal and constitute an important global health care problem. Some key factors that make chronic wounds challenging to heal are attributed to the incessant release of free radicals, which activate the inflammatory system and impair the repair of the wound. Intrinsic characteristics of hydrogels are beneficial for wound healing, but the effective control of free radical levels in the wound and subsequent inflammation is still a challenge. Catechol, the key molecule responsible for the mechanism of adhesion of mussels, has been proven to be an excellent radical scavenger and anti-inflammatory agent. Our approach in this work lies in the preparation of a hybrid system combining the beneficial properties of hydrogels and catechol for its application as a bioactive wound dressing to assist in the treatment of chronic wounds. The hydrogel backbone is obtained through a self-covalent crosslinking between chitosan (Ch) and oxidized hyaluronic acid (HAox) in the presence of a synthetic catechol terpolymer, which is subsequently coordinated to Fe to obtain an interpenetrated polymer network (IPN). The structural analysis, catechol release profiles, in vitro biological behavior and in vivo performance of the IPN are analyzed and compared with the semi-IPN (without Fe) and the Ch/HAox crosslinked hydrogels as controls. Catechol-containing hydrogels present high tissue adhesion strength under wet conditions, support growth, migration and proliferation of hBMSCs, protect cells against oxidative stress damage induce by ROS, and promote down-regulation of the pro-inflammatory cytokine IL-1ß. Furthermore, in vivo experiments reveal their biocompatibility and stability, and histological studies indicate normal inflammatory responses and faster vascularization, highlighting the performance of the IPN system. The novel IPN design also allows for the in situ controlled and sustained delivery of catechol. Therefore, the developed IPN is a suitable ECM-mimic platform with high cell affinity and bioactive functionalities that, together with the controlled catechol release, will enhance the tissue regeneration process and has a great potential for its application as wound dressing.

In Situ Cross-Linkable Polymer Systems and Composites for Osteochondral Regeneration.

Injectable hydrogels have demonstrated being a promising strategy for cartilage and bone tissue engineering applications, owing to their minimal invasive injection procedure, easy incorporation of cells and bioactive molecules, improved contact with the surrounding tissues and ability to match defects with complex irregular shapes, characteristics of osteoarthritic pathology. These unique properties make them highly suitable bioscaffolds for treating defects which are otherwise not easily accessible without and invasive surgical procedure. In this book chapter it has been summarized the novel appropriate injectable hydrogels for cartilage and bone tissue engineering applications of the last few years, including the most commonly used materials for the preparation, both natural and synthetic, and their fabrication techniques. The design of a suitable injectable hydrogel with an adequate gelation time that gathers perfect bioactive, biocompatible, biodegradable and good mechanical properties for clinical repair of damaged cartilage and bone tissue is a challenge of significant medical interest that remain to be achieved.

Bioactive and Bioadhesive Catechol Conjugated Polymers for Tissue Regeneration.

The effective treatment of chronic wounds constitutes one of the most common worldwide healthcare problem due to the presence of high levels of proteases, free radicals and exudates in the wound, which constantly activate the inflammatory system, avoiding tissue regeneration. In this study, we describe a multifunctional bioactive and resorbable membrane with in-built antioxidant agent catechol for the continuous quenching of free radicals as well as to control inflammatory response, helping to promote the wound-healing process. This natural polyphenol (catechol) is the key molecule responsible for the mechanism of adhesion of mussels providing also the functionalized polymer with bioadhesion in the moist environment of the human body. To reach that goal, synthesized statistical copolymers of N-vinylcaprolactam (V) and 2-hydroxyethyl methacrylate (H) have been conjugated with catechol bearing hydrocaffeic acid (HCA) molecules with high yields. The system has demonstrated good biocompatibility, a sustained antioxidant response, an anti-inflammatory effect, an ultraviolet (UV) screen, and bioadhesion to porcine skin, all of these been key features in the wound-healing process. Therefore, these novel mussel-inspired materials have an enormous potential for application and can act very positively, favoring and promoting the healing effect in chronic wounds.