Biology subjects

Justin, A. W.

Publications and source records attributed to Justin, A. W..

6 recordsLinked to original sources

bioRxiv2026

Evaluation of Densified Collagen Tubes as an Interposition Graft in Abdominal Aorta

A major challenge in vascular tissue engineering is the development of scalable fabrication methods capable of producing vascular grafts with suitable mechanical properties and hemocompatibility for both preclinical evaluation and future clinical translation. Using a recently developed method to generate acellular vascular grafts from densified collagen hydrogel, we present a proof-of-concept in vivo evaluation of the scalable collagen graft technology. Thick-walled, genipin-crosslinked grafts, representative of a fabrication approach applicable to clinically relevant vessel sizes, were implanted as interposition grafts in the rat abdominal aorta. These grafts supported rapid neointima formation and re-endothelialisation of the luminal surface but developed late thrombosis, highlighting the challenges of evaluating clinically scalable grafts in small-animal models. To enable assessment of the platform in millimetre-scale vessels, we developed a thin-walled (~100 {micro}m), non-crosslinked graft using a constrained densification approach and hyaluronic acid-mediated modulation of collagen fibril assembly to improve compliance matching and hemocompatibility, with the latter providing a matrix-level approach that may complement surface-bound antithrombotic coatings. These grafts remained patent for up to 17 weeks and exhibited neointima formation, re-endothelialisation, and early cellular infiltration into the graft wall. Together, these findings demonstrate in vivo functionality of densified collagen vascular grafts and establish a foundation for future evaluation in larger animal models.

bioengineering↗

bioRxiv2026

Ventricular Forebrain Organoids Reproduce Macroscale Geometry of the Developing Telencephalon

During development, the size of the neuroepithelial cell pool plays a key role in establishing brain size, determining the numbers of derived progenitors and subsequent neuronal cell types. While early histogenesis is well modelled in brain organoids, the organ-scale geometry of the telencephalon is not accurately recapitulated. Herein, we present a new approach for generating ventral and dorsal forebrain organoids which develop a large ventricular neuroepithelium, characteristic of the closed telencephalic vesicle. Using a growth medium that supports aerobic glycolysis and is typically used for endothelial cells, we modulate neuroepithelial expansion to induce a more anatomically accurate neuroepithelial layer which, upon maturation, thickens physiologically to generate the typical neurogenic layered architecture. In addition, we present a new method for embedding organoids in miniature collagen spheres which mimics native extracellular matrix, stabilizes the ventricular geometry for dynamic culture conditions, and provides a means for incorporating vascular cells for neurovascular development. Finally, we demonstrate that human organoids grown under these conditions exhibit dramatically enlarged ventricles and delayed maturation compared to mouse. Together, this approach provides a model of the forebrain neuroepithelium with morphogenetic macroscale geometry and tissue architecture, suitable for investigating neurodevelopment and disease.

bioengineering↗

bioRxiv2026

Multiscale computational framework for generating vascularizedbiohybrid tissue constructs

Vascularization remains a central challenge in building large-scale biohybrid tissues that integrate living and synthetic components. Without a perfusable vascular network, nutrient delivery and waste removal become insufficient, leading to hypoxia and a loss of viability in thicker tissue constructs. We present Lattice Sequence Vascularization (LSV), a multiscale computational design framework for generating hierarchical, biomimetic vascular networks that are compatible with 3D-printing constraints and manufacturable within arbitrary geometries. LSV employs a divide-and-conquer strategy in which vessels grow and remodel at a specified terminal scale before recursively subdividing to form the full hierarchy. By enforcing hierarchy, LSV produces networks that exhibit self-similarity across length scales, a defining feature of physiological vasculature. The framework integrates synthetic considerations (e.g., hydrogel permeability), biological constraints (Murrays law, cross-scale biomimicry, organ-specific perfusion requirements) and manufacturing requirements relevant to 3D printing and microfabrication. We demonstrate the incorporation of capillary-scale functional substructures (e.g. organoid traps) and the generation of complex architectures with multiple inlets and outlets (e.g. liver-like geometries), enabling organ-scale vasculature design.

bioengineering↗

bioRxiv2024

A chemically defined and xeno-free hydrogel system for regenerative medicine.

Biofabricated scaffolds facilitate bona fide cellular interactions, cell type specification, and the formation of three-dimensional tissue architecture from human pluripotent stem cells (hPSCs). However, xenogenic biomaterials are poorly defined, and synthetic biomaterials remain underdeveloped and understudied, hindering regulatory approval for clinical use and preventing the translation of lab-grown therapies. Here, we describe a protein screen-based hydrogel system biofabricated from clinical-grade human components. We show that "Alphagel", a base hydrogel comprising human embryonic matrices, supports the trilineage differentiation of hPSCs into neural, cardiac, and liver tissue. Alphagel is also shown to be biocompatible and biodegradable in vivo. Further, upon adding select proteins from maturing human foetal liver to Alphagel, we show that the resulting hydrogel (termed "Hepatogel") enhances the differentiation of hPSC-derived hepatocytes (H-iHeps) compared with Matrigel. Importantly, when injected into mice livers, Hepatogel significantly improves the retention of H-iHeps compared to standard aqueous cell injections. Altogether, our results provide proof of concept that customisable and organ-specific hydrogel systems are a valuable tool for developing clinically translatable therapies for regenerative medicine and tissue engineering.

bioengineering↗

bioRxiv2021

Densified Collagen Tubular Grafts for Human Tissue Replacement and Disease Modelling Applications

There is a significant need across multiple indications for an off-the-shelf bioengineered tubular graft which fulfils the mechanical and biological requirements for implantation and function but does not necessarily require cells for manufacture or deployment. Herein, we present a tissue-like tubular construct using a cell-free, materials-based method of manufacture, utilizing densified collagen hydrogel. Our tubular grafts are seamless, mechanically strong, customizable in terms of lumen diameter and wall thickness, and display a uniform fibril density across the wall thickness and along the tube length. While the method enables acellular grafts to be generated rapidly, inexpensively, and to a wide range of specifications, the cell-compatible densification process also enables a high density of cells to be incorporated uniformly into the walls of the tubes, which we show can be maintained under perfusion culture. Additionally, the method enables tubes consisting of distinct cell domains with cellular configurations at the boundaries which may be useful for modelling aortic disease. Further, we demonstrate additional steps which allow for luminal surface patterning. These results highlight the universality of this approach and its potential for developing the next generation of bioengineered grafts.

bioengineering↗

bioRxiv2021

A vascularized tumoroid model for studying human glioblastoma angiogenesis

Glioblastoma (GBM) angiogenesis is critical for tumor growth and recurrence, making it a compelling therapeutic target. Here, a disease-relevant, vascularized tumoroid in vitro model with stem-like features and stromal surrounds is reported. The model is used to recapitulate how individual components of the GBMs complex brain microenvironment such as hypoxia, vasculature-related stro-mal cells and growth factors support GBM angiogenesis. It is scalable, tractable, cost-effective and can be used with biologically-derived or biomimetic matrices. Patient-derived primary GBM cells are found to closely participate in blood vessel formation in contrast to a GBM cell line containing differentiated cells. Exogenous growth factors amplify this effect under normoxia but not at hypoxia suggesting that a significant amount of growth factors is already being produced under hypoxic conditions. Under hypoxia, primary GBM cells and umbilical vein endothelial cells are found to strongly co-localize in a mosaic pattern to form sprouting vascular networks, with GBM cells acquiring an endothelial-like behaviour, which has been reported to occur in vivo. These findings demonstrate that our 3D tumoroid in vitro model exhibits biomimetic attributes that may permit its use as a preclinical model in studying microenvironment cues of tumor angiogenesis.

bioengineering↗