Graphene and graphene oxide (GO) structures and their reduced forms, e.g., GO paper and partially or fully reduced three-dimensional (3D) aerogels, are at the forefront of materials design for extensive biomedical applications that allow for the proliferation and differentiation/maturation of cells, drug delivery, and anticancer therapies. Various viability tests that have been conducted in vitro on human cells and in vivo on mice reveal very promising results, which make graphene-based materials suitable for real-life applications. In this review, we will give an overview of the latest studies that utilize graphene-based structures and their composites in biological applications and show how the biomimetic behavior of these materials can be a step forward in bridging the gap between nature and synthetically designed graphene-based nanomaterials.

This review summarizes recent progress in the development and applications of metal–organic gels (MOGs) and their hybrids and derivatives dividing them into subclasses and discussing their synthesis, design and structure–property relationship.

Cartilage engineering with stem cells in 3D scaffolds is a promising future therapy to treat cartilage defects. One challenge in the field is to design carriers to efficaciously deliver biological factors in 3D scaffolds containing stem cells to appropriately guide differentiation of these cells in same scaffolds and promote specific tissue synthesis. Graphene-based 2D nanomaterials have recently attracted extensive interest for their biomedical applications as they can adsorb a plethora of biological molecules, thus offering high potential as delivery carriers. This study utilized graphene oxide (GO) flakes to adsorb transforming growth factor β3 (TGF-β3), which were then incorporated into a collagen hydrogel. Human mesenchymal stem cells (hMSCs) were encapsulated in the same gel and chondrogenic differentiation assessed. The study showed GO flakes adsorbed > 99% TGF-β3 with <1.7% release. Adsorbed TGF-β3 retained a similar conformation to its dissolved counterpart (free protein) but importantly demonstrated greater conformational stability. Smad2 phosphorylation was promoted, and higher chondrogenic gene expression and cartilage-specific extracellular matrix deposition were achieved compared to exogenously delivering TGF-β3 in culture media. Effects were sustained in long-term 28-day culture. The results demonstrate GO flakes as highly-efficient for delivering GFs in 3D to guide cells in the same scaffold and induce tissue formation. The ability of GO flakes to provide sustained local delivery makes this material attractive for tissue engineering strategies, in particular for regionally-specific MSC differentiation (e.g. osteochondral tissue engineering). STATEMENT OF SIGNIFICANCE: Cartilage engineering involving stem cells in 3D scaffolds is a promising future therapy to treat cartilage defects which can lead to debilitating conditions such as osteoarthritis. However, this field faces the challenge to design delivery carriers to efficaciously deliver biological factors inside these 3D cell-containing scaffolds for appropriately-guided cell differentiation. Graphene-based 2D nanomaterials offer high potential as delivery carriers, but to date studies using them to deliver biological factors have been restricted to 2D substrates, non-scaffold cell masses, or acellular 3D scaffolds. Our study for the first time demonstrated simultaneously incorporating both human mesenchymal stem cells (hMSCs) and GO (graphene oxide)-adsorbed growth factor TGFβ3 into a 3D scaffold, where GO-adsorbed TGFβ3 enhanced chondrogenic differentiation of hMSCs and cartilage-tissue synthesis throughout the scaffold without needing to repeatedly supply TGFβ3 exogenously.

Cell-based therapies have shown significant promise in tissue engineering with one key challenge being the delivery and retention of cells. As a result, significant efforts have been made in the past decade to design injectable biomaterials to host and deliver cells at injury sites. Intervertebral disc (IVD) degeneration, a major cause of back pain, is a particularly relevant example where a minimally-invasive cellular therapy could bring significant benefits specifically at the early stages of the disease, when a cell-driven process starts in the gelatinous core of the IVD, the nucleus pulposus (NP). In this present study we explore the use of graphene oxide (GO) as nano-filler for the reinforcement of FEFKFEFK (β-sheet forming self-assembling peptide) hydrogels. Our results confirm the presence of strong interactions between FEFKFEFK and GO flakes with the peptide coating and forming short thin fibrils on the surface of the flakes. These strong interactions were found to affect the bulk properties of hybrid hydrogels. At pH 4 electrostatic interactions between the peptide fibres and the peptide-coated GO flakes are thought to govern the final bulk properties of the hydrogels while at pH 7, after conditioning with cell culture media, electrostatic interactions are removed leaving the hydrophobic interactions to govern hydrogel final properties. The GO-F820 hybrid hydrogel, with mechanical properties similar to the NP, was shown to promote high cell viability and retained cell metabolic activity in 3D over the 7 days of culture and therefore shown to harbour significant potential as an injectable hydrogel scaffold for the in-vivo delivery of NP cells. STATEMENT OF SIGNIFICANCE: Short self-assembling peptide hydrogels (SAPHs) have attracted significant interest in recent years as they can mimic the natural extra-cellular matrix, holding significant promise for the ab initio design of cells' microenvironments. Recently the design of hybrid hydrogels for biomedical applications has been explored through the incorporation of specific nanofillers. In this study we exploited graphene oxide (GO) as nanofiller to design hybrid injectable 3Dscaffolds for the delivery of nucleus pulposus cells (NPCs) for intervertebral disc regeneration. Our work clearly shows the presence of strong interactions between peptide and GO, mimicking the mechanical properties of the NP tissue and promoting high cell viability and metabolic activity. These hybrid hydrogels therefore harbour significant potential as injectable scaffolds for the in vivo delivery of NPCs.

Two-dimensional (2D) materials remain highly interesting for assembling three-dimensional (3D) structures, amongst others, in the form of macroscopic hydrogels. Herein, we present a novel approach for inducing chemical inter-sheet crosslinks via an ethylenediamine mediated reaction between Ti₃C₂T_x and graphene oxide in order to obtain a reduced graphene oxide-MXene (rGO-MXene) hydrogel. The composite hydrogels are hydrophilic with a stiffness of ~20 kPa. They also possess a unique inter-connected porous architecture, which led to a hitherto unprecedented ability of human cells across three different types, epithelial adenocarcinoma, neuroblastoma and fibroblasts, to form inter-connected three-dimensional networks. The attachments of the cells to the rGO-MXene hydrogels were superior to those of the sole rGO-control gels. This phenomenon stems from the strong affinity of cellular protrusions (neurites, lamellipodia and filopodia) to grow and connect along architectural network paths within the rGO-MXene hydrogel, which could lead to advanced control over macroscopic formations of cellular networks for technologically relevant bioengineering applications, including tissue engineering and personalized diagnostic networks-on-chip. STATEMENT OF SIGNIFICANCE: Conventional hydrogels are made of interconnected polymeric fibres. Unlike conventional case, we used hydrothermal and chemical approach to form interconnected porous hydrogels made of two-dimensional flakes from graphene oxide and metal carbide from a new family of MXenes (Ti₃C₂T_x). This way, we formed three-dimensional porous hydrogels with unique porous architecture of well-suited chemical surfaces and stiffness. Cells from three different types cultured on these scaffolds formed extended three-dimensional networks - a feature of extended cellular proliferation and pre-requisite for formation of organoids. Considering the studied 2D materials typically constitute materials exhibiting enhanced supercapacitor performances, our study points towards better understanding of design of tissue engineering materials for the future bioengineering fields including personalized diagnostic networks-on-chip, such as artificial heart actuators.