Human lung organoids develop into adult airway-like structures directed by physico-chemical biomaterial properties.

Tissues derived from human pluripotent stem cells (hPSCs) often represent early stages of fetal development, but mature at the molecular and structural level when transplanted into immunocompromised mice. hPSC-derived lung organoids (HLOs) transplantation has been further enhanced with biomaterial scaffolds, where HLOs had improved tissue structure and cellular differentiation. Here, our goal was to define the physico-chemical biomaterial properties that maximally enhanced transplant efficiency, including features such as the polymer type, degradation, and pore interconnectivity of the scaffolds. We found that transplantation of HLOs on microporous scaffolds formed from poly (ethylene glycol) (PEG) hydrogel scaffolds inhibit growth and maturation, and the transplanted HLOs possessed mostly immature lung progenitors. On the other hand, HLOs transplanted on poly (lactide-co-glycolide) (PLG) scaffolds or polycaprolactone (PCL) led to tube-like structures that resembled both the structure and cellular diversity of an adult airway. Our data suggests that scaffold pore interconnectivity and polymer degradation contributed to the maturation, and we found that the size of the airway structures and the total size of the transplanted tissue was influenced by the material degradation rate. Collectively, these biomaterial platforms provide a set of tools to promote maturation of the tissues and to control the size and structure of the organoids.

Microporous scaffolds support assembly and differentiation of pancreatic progenitors into ß-cell clusters.

Human pluripotent stem cells (hPSCs) represent a promising cell source for the development of ß-cells for use in therapies for type 1 diabetes. Current culture approaches provide signals to mimic a temporal control of organogenesis to drive the differentiation towards ß-cells. However, spatial control may represent an opportunity to improve the efficiency and manufacturing of ß-cells. Herein, we adapted the current culture systems to microporous biomaterials with the hypothesis that the pores can guide the assembly of pancreatic progenitors into clusters of defined size that can influence maturation. The scaffold culture allowed hPSC-derived pancreatic progenitors to form clusters at a consistent size as cells differentiated. By modulating the scaffold pore sizes, we observed 250-425 µm pore size scaffold cultures augmented insulin expression and key ß-cell maturation markers compared to cells cultured in suspension. Furthermore, when compared to suspension cultures, the scaffold culture showed increased insulin secretion in response to glucose stimulus indicating the development of functional ß-cells. In addition, scaffolds facilitated cell-cell interactions enabled by the scaffold design and supported cell-mediated matrix deposition of extracellular matrix (ECM) proteins associated with the basement membrane of islet cells. We further investigated the influence of ECM on cell development by incorporating an ECM matrix on the scaffold prior to cell seeding; however, their presence did not further enhance maturation. These results suggest the microporous scaffold culture provides a conducive environment that drives in vitro differentiation of hPSC-derived insulin-producing glucose-responsive ß-cells and demonstrates the feasibility of these scaffolds as a biomanufacturing platform. STATEMENT OF SIGNIFICANCE: Cell therapy for diabetes is a promising strategy, yet generating limitless insulin-producing mature ß-cells from human pluripotent stem cells (hPSCs) remains a challenge. Current hPSC differentiation methods involve media containing signals to drive maturation toward ß-cells and spontaneous cluster formation. Herein, we sought to provide spatial cues to guide assembly of cells into 3D structures by culture within the pores of a microporous scaffold. The scaffolds direct cell-cell interactions within the pores and provide a support for cell-mediated matrix deposition that collectively creates a niche to promote functional hPSC-derived ß-cell clusters. These scaffolds for 3D culture may contribute to hPSC differentiation methods for the generation of ß-cells that can treat patients with diabetes.

It's All in the Delivery: Designing Hydrogels for Cell and Non-viral Gene Therapies.

Hydrogels provide a regenerative medicine platform with their ability to create an environment that supports transplanted or endogenous infiltrating cells and enables these cells to restore or replace the function of tissues lost to disease or trauma. Furthermore, these systems have been employed as delivery vehicles for therapeutic genes, which can direct and/or enhance the function of the transplanted or endogenous cells. Herein, we review recent advances in the development of hydrogels for cell and non-viral gene delivery through understanding the design parameters, including both physical and biological components, on promoting transgene expression, cell engraftment, and ultimately cell function. Furthermore, this review identifies emerging opportunities for combining cell and gene delivery approaches to overcome challenges to the field.

Gelation chemistries for the encapsulation of nanoparticles in composite gel microparticles for lung imaging and drug delivery.

The formation of 10-40 µm composite gel microparticles (CGMPs) comprised of ∼100 nm drug containing nanoparticles (NPs) in a poly(ethylene glycol) (PEG) gel matrix is described. The CGMP particles enable targeting to the lung by filtration from the venous circulation. UV radical polymerization and Michael addition polymerization reactions are compared as approaches to form the PEG matrix. A fluorescent dye in the solid core of the NP was used to investigate the effect of reaction chemistry on the integrity of encapsulated species. When formed via UV radical polymerization, the fluorescence signal from the NPs indicated degradation of the encapsulated species by radical attack. The degradation decreased fluorescence by 90% over 15 min of UV exposure. When formed via Michael addition polymerization, the fluorescence was maintained. Emulsion processing using controlled shear stress enabled control of droplet size with narrow polydispersity. To allow for emulsion processing, the gelation rate was delayed by adjusting the solution pH. At a pH = 5.4, the gelation occurred at 3.5 h. The modulus of the gels was tuned over the range of 5 to 50 kPa by changing the polymer concentration between 20 and 70 vol %. NP aggregation during polymerization, driven by depletion forces, was controlled by the reaction kinetics. The ester bonds in the gel network enabled CGMP degradation. The gel modulus decreased by 50% over 27 days, followed by complete gel degradation after 55 days. This permits ultimate clearance of the CGMPs from the lungs. The demonstration of uniform delivery of 15.8 ± 2.6 µm CGMPs to the lungs of mice, with no deposition in other organs, is shown, and indicates the ability to concentrate therapeutics in the lung while avoiding off-target toxic exposure.