Recent advancements in 3D bioprinting have led to the fabrication of more complex, more precise, and larger printed tissue constructs. As the field continues to advance, it is critical to develop quantitative benchmarks to compare different bio-inks for key cell-biomaterial interactions, including (1) cell sedimentation within the ink cartridge, (2) cell viability during extrusion, and (3) cell viability after ink curing. Here we develop three simple protocols for quantitative analysis of bio-ink performance. These methods are used to benchmark the performance of two commonly used bio-inks, poly(ethylene glycol) diacrylate (PEGDA) and gelatin methacrylate (GelMA), against three formulations of a novel bio-ink, Recombinant-protein Alginate Platform for Injectable Dual-crosslinked ink (RAPID ink). RAPID inks undergo peptide-self-assembly to form weak, shear-thinning gels in the ink cartridge and undergo electrostatic crosslinking with divalent cations during curing. In the one-hour cell sedimentation assay, GelMA, the RAPID inks, and PEGDA with xanthan gum prevented appreciable cell sedimentation, while PEGDA alone or PEGDA with alginate experienced significant cell settling. To quantify cell viability during printing, 3T3 fibroblasts were printed at a constant flowrate of 75 μl/min and immediately tested for cell membrane integrity. Less than 10% of cells were damaged using the PEGDA and GelMA bio-inks, while less than 4% of cells were damaged using the RAPID inks. Finally, to evaluate cell viability after curing, cells were exposed to ink-specific curing conditions for five minutes and tested for membrane integrity. After exposure to light with photo-initiator at ambient conditions, over 50% of cells near the edges of printed PEGDA and GelMA droplets were damaged. In contrast, fewer than 20% of cells found near the edges of RAPID inks were damaged after a 5-minute exposure to curing in a 10 mM CaCl2 solution. As new bio-inks continue to be developed, these protocols offer a convenient means to quantitatively benchmark their performance against existing inks.

NATure BIoMedIcAl eNgINeerINgdeveloped an approach for in-cell site-specific protein phosphorylation to synthesize bioactive proteins fused with a phosphorylated alum-binding peptide (ABP) tag. We used this approach to produce a series of ABP-labelled cytokines, which rapidly adsorbed to alum after simple mixing, and upon i.t. injection were retained in tumours for more than a week. Applied to the cytokine IL-12, this approach dramatically increased i.t. retention of the cytokine and eliminated systemic toxicities seen upon i.t. injection of the free drug, while also increasing anti-tumour efficacy. Moreover, a single i.t. dose of alum-anchored IL-12 elicited strong IFN-γ-dependent collaboration between innate and adaptive immune cells, producing robust systemic anti-tumour responses in multiple poorly immunogenic preclinical models when combined with systemic checkpoint blockade therapy. ResultsTargeted phosphorylation via an in-cell approach is robust. A single kinase, Fam20C, is responsible for phosphorylation of

Fiber drawing enables scalable fabrication of multifunctional flexible fibers that integrate electrical, optical, and microfluidic modalities to record and modulate neural activity. Constraints on thermomechanical properties of materials, however, have prevented integrated drawing of metal electrodes with low‐loss polymer waveguides for concurrent electrical recording and optical neuromodulation. Here, two fabrication approaches are introduced: 1) an iterative thermal drawing with a soft, low melting temperature (Tm) metal indium, and 2) a metal convergence drawing with traditionally non‐drawable high Tm metal tungsten. Both approaches deliver multifunctional flexible neural interfaces with low‐impedance metallic electrodes and low‐loss waveguides, capable of recording optically‐evoked and spontaneous neural activity in mice over several weeks. These fibers are coupled with a light‐weight mechanical microdrive (1 g) that enables depth‐specific interrogation of neural circuits in mice following chronic implantation. Finally, the compatibility of these fibers with magnetic resonance imaging is demonstrated and they are applied to visualize the delivery of chemical payloads through the integrated channels in real time. Together, these advances expand the domains of application of the fiber‐based neural probes in neuroscience and neuroengineering.

Supramolecular hyaluronic acid hydrogels formed via 2 : 1 homoternary complexes of coumarin and cucurbit[8]uril can reversibly toggle between physical and covalent states.

The clinical outcomes and 5-year survival rate for patients with glioblastoma (GB) make it among the most pernicious and challenging diseases to treat. Despite all the resources, time, and talent focused on developing targeted and/or local delivery technologies by the biomaterials community for GB, the clinical performance of the FDA-approved therapy carmustine ((BCNU)-loaded polyanhydride wafers) and clinical trials of other material approaches have been discouraging. As disappointing is the remarkably stagnant clinical translation of next-generation material approaches for GB. Despite encouraging preclinical results from hydrogels and modified wafer formulations loaded with more efficacious chemotherapies, a total of zero have completed even a phase I clinical trial. Other strategies, including convection-enhanced delivery, microsphere formulations, or drug-loaded nanoparticles have seen limited, albeit some, translation into the clinic with mixed results. This lackluster progress can be attributed, in part, to the paucity of communication between material scientists, biomedical scientists, and clinicians. When examining the purported clinical relevance of embedding certain material properties into formulations, it is clear that some widely known truths about the nature of GB progression among clinicians have not reached the biomaterials community.Furthermore, a closer examination of the lessons from the BCNU wafers and other clinical trials of GB drug delivery materials may enrich and inspire materials scientists to create new systems that satisfy unmet medical needs identified by the clinical community. In tandem, clinicians and biomedical scientists may benefit from a short review highlighting the biocompatibility, safety, longevity, kinetics, tunability, and efficacy of promising new drug delivery materials without inundation by chemical and physical characterizations or discussions.Another key challenge in treating GB is an incomplete understanding of disease pathophysiology, such as mechanisms driving intrinsic and adaptive GB cell chemoresistance. A combined approach where biomedical scientists and material To date, the clinical outcomes and survival rates for patients with glioblastoma (GB) remain poor. A promising approach to disease-modification involves local delivery of adjuvant chemotherapy into the resection cavity, thus circumventing the restrictions imposed by the blood-brain barrier. The clinical performance of the only FDA-approved local therapy for GB [carmustine (BCNU)-loaded polyanhydride wafers], however, has been disappointing. There is an unmet medical need in the local treatment of GB for drug delivery vehicles that provide sustained local release of small molecules and combination drugs over several months. Herein, key quantitative lessons from the use of local and systemic adjuvant chemotherapy for GB in the clinic are outlined, and it is discussed how these can inform the development of next-generation therapies. Several recent approaches are highlighted, and it is proposed that long...

Direct injection of therapies into tumors has emerged as an administration route capable of achieving high local drug exposure and strong anti-tumor response. A diverse array of immune agonists ranging in size and target are under development as local immunotherapies. However, due to the relatively recent adoption of intratumoral administration, the pharmacokinetics of locally-injected biologics remains poorly defined, limiting rational design of tumor-localized immunotherapies. Here we define a pharmacokinetic framework for biologics injected intratumorally that can predict tumor exposure and effectiveness. We find empirically and computationally that extending the tumor exposure of locally-injected interleukin-2 by increasing molecular size and/or improving matrix-targeting affinity improves therapeutic efficacy in mice. By tracking the distribution of intratumorally-injected proteins using positron emission tomography, we observe size-dependent enhancement in tumor exposure occurs by slowing the rate of diffusive escape from the tumor and by increasing partitioning to an apparent viscous region of the tumor. In elucidating how molecular weight and matrix binding interplay to determine tumor exposure, our model can aid in the design of intratumoral therapies to exert maximal therapeutic effect.

Synthetic hydrogels are an important class of materials in tissue engineering, drug delivery, and other biomedical fields. Their mechanical and electrical properties can be tuned to match those of biological tissues. In this work, hydrogels that exhibit both mechanical and electrical biomimicry are reported. The presented dual networks consist of supramolecular networks formed from 2:1 homoternary complexes of imidazolium‐based guest molecules in cucubit[8]uril and covalent networks of oligoethylene glycol‐(di)methacrylate. The viscoelastic properties of human brain tissues are also investigated. The mechanical properties of the dual network gels are benchmarked against the human tissue, and it is found that they both are neuro‐mimetic and exhibit cytocompatibility in a neural stem cell model.

The scalable production of uniformly distributed graphene (GR)-based composite materials remains a sizable challenge. While GR-polymer nanocomposites can be manufactured at large scale, processing limitations result in poor control over the homogeneity of hydrophobic GR sheets in the matrices. Such processes often result in difficulties controlling stability and avoiding aggregation, therefore eliminating benefits that might have otherwise arisen from the nanoscopic dimensions of GR. Here, we report an exfoliated and stabilized GR dispersion in water. Cucurbit[8]uril (CB[8])-mediated hostguest chemistry was used to obtain supramolecular hydrogels consisting of uniformly distributed GR and guest-functionalized macromolecules. The obtained GR-hydrogels show superior bioelectrical properties over identical systems produced without CB[8]. Utilizing such supramolecular interactions with biologically-derived macromolecules is a promising approach to stabilize graphene in water and avoid oxidative chemistry.