Inflammation-induced subcutaneous neovascularization for the long-term survival of encapsulated islets without immunosuppression.

Cellular therapies for type-1 diabetes can leverage cell encapsulation to dispense with immunosuppression. However, encapsulated islet cells do not survive long, particularly when implanted in poorly vascularized subcutaneous sites. Here we show that the induction of neovascularization via temporary controlled inflammation through the implantation of a nylon catheter can be used to create a subcutaneous cavity that supports the transplantation and optimal function of a geometrically matching islet-encapsulation device consisting of a twisted nylon surgical thread coated with an islet-seeded alginate hydrogel. The neovascularized cavity led to the sustained reversal of diabetes, as we show in immunocompetent syngeneic, allogeneic and xenogeneic mouse models of diabetes, owing to increased oxygenation, physiological glucose responsiveness and islet survival, as indicated by a computational model of mass transport. The cavity also allowed for the in situ replacement of impaired devices, with prompt return to normoglycemia. Controlled inflammation-induced neovascularization is a scalable approach, as we show with a minipig model, and may facilitate the clinical translation of immunosuppression-free subcutaneous islet transplantation.

A predictive computational platform for optimizing the design of bioartificial pancreas devices.

The delivery of encapsulated islets or stem cell-derived insulin-producing cells (i.e., bioartificial pancreas devices) may achieve a functional cure for type 1 diabetes, but their efficacy is limited by mass transport constraints. Modeling such constraints is thus desirable, but previous efforts invoke simplifications which limit the utility of their insights. Herein, we present a computational platform for investigating the therapeutic capacity of generic and user-programmable bioartificial pancreas devices, which accounts for highly influential stochastic properties including the size distribution and random localization of the cells. We first apply the platform in a study which finds that endogenous islet size distribution variance significantly influences device potency. Then we pursue optimizations, determining ideal device structures and estimates of the curative cell dose. Finally, we propose a new, device-specific islet equivalence conversion table, and develop a surrogate machine learning model, hosted on a web application, to rapidly produce these coefficients for user-defined devices.

Engineering a Hierarchical Biphasic Gel for Subcutaneous Vascularization.

Implanted cell-containing grafts require a robust and functional vasculature to supply oxygen and nutrients, as well as clear metabolic waste products. However, it remains challenging to fabricate tunable, vascular-promoting scaffolds without incorporating additional biologics. Here, a biphasic gel consisting of a highly porous aerogel and a degradable fibrin hydrogel for inducing vascularization is presented. The highly porous (>90%) and stable aerogel is assembled from short microfibers by being dispersed in an aqueous solution that can be 3D printed into various configurations. The biphasic gel demonstrates good compression-resistance: 70.30% Young's modulus is recovered over 20 cycles of 65% compression under water. Furthermore, it is confirmed that tissue cells and blood vessels can penetrate a thick (≈3 mm) biphasic gel in the subcutaneous space of mice. Finally, the biphasic gel doubles the vascular ingrowth compared to a composite of a commercial surgical polyester felt and a fibrin hydrogel upon subcutaneous implantation in mice after 4 weeks. The design of this biphasic gel may advance the development of vascularized scaffolds.

A Safe, Fibrosis-Mitigating, and Scalable Encapsulation Device Supports Long-Term Function of Insulin-Producing Cells.

Encapsulation and transplantation of insulin-producing cells offer a promising curative treatment for type 1 diabetes (T1D) without immunosuppression. However, biomaterials used to encapsulate cells often elicit foreign body responses, leading to cellular overgrowth and deposition of fibrotic tissue, which in turn diminishes mass transfer to and from transplanted cells. Meanwhile, the encapsulation device must be safe, scalable, and ideally retrievable to meet clinical requirements. Here, a durable and safe nanofibrous device coated with a thin and uniform, fibrosis-mitigating, zwitterionically modified alginate hydrogel for encapsulation of islets and stem cell-derived beta (SC-ß) cells is reported. The device with a configuration that has cells encapsulated within the cylindrical wall, allowing scale-up in both radial and longitudinal directions without sacrificing mass transfer, is designed. Due to its facile mass transfer and low level of fibrotic reactions, the device supports long-term cell engraftment, correcting diabetes in C57BL6/J mice with rat islets for up to 399 days and SCID-beige mice with human SC-ß cells for up to 238 days. The scalability and retrievability in dogs are further demonstrated. These results suggest the potential of this new device for cell therapies to treat T1D and other diseases.

A nanofibrous encapsulation device for safe delivery of insulin-producing cells to treat type 1 diabetes.

Transplantation of stem cell-derived ß (SC-ß) cells represents a promising therapy for type 1 diabetes (T1D). However, the delivery, maintenance, and retrieval of these cells remain a challenge. Here, we report the design of a safe and functional device composed of a highly porous, durable nanofibrous skin and an immunoprotective hydrogel core. The device consists of electrospun medical-grade thermoplastic silicone-polycarbonate-urethane and is soft but tough (~15 megapascal at a rupture strain of >2). Tuning the nanofiber size to less than ~500 nanometers prevented cell penetration while maintaining maximum mass transfer and decreased cellular overgrowth on blank (cell-free) devices to as low as a single-cell layer (~3 micrometers thick) when implanted in the peritoneal cavity of mice. We confirmed device safety, indicated as continuous containment of proliferative cells within the device for 5 months. Encapsulating syngeneic, allogeneic, or xenogeneic rodent islets within the device corrected chemically induced diabetes in mice and cells remained functional for up to 200 days. The function of human SC-ß cells was supported by the device, and it reversed diabetes within 1 week of implantation in immunodeficient and immunocompetent mice, for up to 120 and 60 days, respectively. We demonstrated the scalability and retrievability of the device in dogs and observed viable human SC-ß cells despite xenogeneic immune responses. The nanofibrous device design may therefore provide a translatable solution to the balance between safety and functionality in developing stem cell-based therapies for T1D.

Hydrogels in Emerging Technologies for Type 1 Diabetes.

Type 1 diabetes therapies that afford tighter glycemic control in a more manageable and painless manner for patients has remained a central focus of next-generation diabetes therapies. In many of these emerging technologies, namely, self-regulated insulin delivery and cell replacement therapies, hydrogels are employed to mitigate some of the most long-standing challenges. In this Review, we summarize recent developments in the use of hydrogels for both insulin delivery and insulin-producing cell therapies for type 1 diabetes management. We first outline perspectives in glucose sensitive hydrogels for smart insulin delivery, pH sensitive polymeric hydrogels for oral insulin delivery, and other physiochemical signals used to trigger insulin release from hydrogels. We, then, investigate the use of hydrogels in the encapsulation of insulin secreting cells with a special emphasis on hydrogels designed to mitigate the foreign body response, provide a suitable extracellular microenvironment, and improve mass transfer through oxygen supplementation and vascularization. Evaluations of limitations and promising directions for future research are also considered. Continuing interdisciplinary and collaborative research efforts will be required to produce hydrogels with instructive biochemical microenvironments necessary to address the enduring challenges of emerging type 1 diabetes therapies.

Microtubules and motor proteins support zebrafish neuronal migration by directing cargo.

Neuronal migration during development is necessary to form an ordered and functional brain. Postmitotic neurons require microtubules and dynein to move, but the mechanisms by which they contribute to migration are not fully characterized. Using tegmental hindbrain nuclei neurons in zebrafish embryos together with subcellular imaging, optogenetics, and photopharmacology, we show that, in vivo, the centrosome's position relative to the nucleus is not linked to greatest motility in this cell type. Nevertheless, microtubules, dynein, and kinesin-1 are essential for migration, and we find that interference with endosome formation or the Golgi apparatus impairs migration to a similar extent as disrupting microtubules. In addition, an imbalance in the traffic of the model cargo Cadherin-2 also reduces neuronal migration. These results lead us to propose that microtubules act as cargo carriers to control spatiotemporal protein distribution, which in turn controls motility. This adds crucial insights into the variety of ways that microtubules can support successful neuronal migration in vivo.

Interconnected Toroidal Hydrogels for Islet Encapsulation.

Islet encapsulation and transplantation promises to improve upon current treatments for type 1 diabetes mellitus, though several limitations remain. Macroscale devices have been designed for in vivo transplantation and retrieval, but traditional geometries do not support clinically adequate mass transfer of nutrients to and insulin from the encapsulated tissue. Microcapsule technologies have improved mass transfer properties, but their clinical translation remains challenging as their complete retrieval is difficult, should the graft become a safety concern. Here, the design, characterization and testing of a novel encapsulation structure, comprised of elastomer-reinforced interconnected toroidal hydrogels is reported. These donut-shaped hydrogels feature a high surface area, higher than conventional spherical capsules of the same volume, bestowing suitable mass transport conditions, while allowing interconnection and reversible deformation for intraperitoneal implantation and retrieval. Diabetes correction up to 12 weeks and complete retrieval is achieved in a diabetic mouse model, providing a proof-of-concept for the potential application as a type 1 diabetes cell replacement therapy.

Nanotechnology in cell replacement therapies for type 1 diabetes.

Islet transplantation is a promising long-term, compliance-free, complication-preventing treatment for type 1 diabetes. However, islet transplantation is currently limited to a narrow set of patients due to the shortage of donor islets and side effects from immunosuppression. Encapsulating cells in an immunoisolating membrane can allow for their transplantation without the need for immunosuppression. Alternatively, "open" systems may improve islet health and function by allowing vascular ingrowth at clinically attractive sites. Many processes that enable graft success in both approaches occur at the nanoscale level-in this review we thus consider nanotechnology in cell replacement therapies for type 1 diabetes. A variety of biomaterial-based strategies at the nanometer range have emerged to promote immune-isolation or modulation, proangiogenic, or insulinotropic effects. Additionally, coating islets with nano-thin polymer films has burgeoned as an islet protection modality. Materials approaches that utilize nanoscale features manipulate biology at the molecular scale, offering unique solutions to the enduring challenges of islet transplantation.