Co-transplantation of Islets-Laden Microgels and Biodegradable O2-Generating Microspheres for Diabetes Treatment.

Pancreatic islets transplantation is an optimal alternative to exogenous insulin injection for long-term effective type 1 diabetes treatment. However, direct islets transplantation without any protection can induce cell necrosis due to severe host immune rejection. Insufficient O2 supply induced by the lack of capillary network at the early stage of islets transplantation is another critical constraint limiting islets survival and insulin-secretion function. In this paper, we design a novel co-transplantation system composed of islets-laden nanocomposite microgels and O2-generating microspheres. In particular, nanocomposite microgels confer the encapsulated islets with simultaneous physical protection and chemical anti-inflammation/immunosuppression by covalently anchoring rapamycin-loaded cyclodextrin nanoparticles to microgel network. Meanwhile, O2-generating microspheres prepared by blending inorganic peroxides in biodegradable polycaprolactone and polylactic acid can generate in situ O2 gas and thus avoid hypoxia environment around transplanted islets. In vivo therapeutic effect of diabetic mice proves the reversion of the high blood glucose level back to normoglycemia and superior glucose tolerance for at least 90 days post co-transplantation. In brief, the localized drug and oxygen codelivery, as well as physical protection provided by our co-transplantation system, has the potential to overcome to a large extent the inflammatory, hypoxia, and host immune rejection after islets transplantation. This new strategy may have wider application in other cell replacement therapies.

Dynamic Nanocomposite Microgel Assembly with Microporosity, Injectability, Tissue-Adhesion, and Sustained Drug Release Promotes Articular Cartilage Repair and Regeneration.

Owing to the lack of blood vessels, nerves, and lymph, articular cartilage defect is difficult to self-repair. Although several cartilage tissue engineering products have been authorized for clinical use, there are still some problems such as large surgical wounds, weak adhesion with the host tissue, and the limited source of autologous chondrocytes. In this paper, a novel dynamic nanocomposite microgel assembly with excellent microporosity, injectability, tissue-adhesion, and sustained kartogenin (KGN) release is reported. Specifically, KGN-loaded cyclodextrin nanoparticles are synthesized through nanoemulsification and incorporated into bone marrow mesenchymal stem cell (BMSCs)-laden microgels via droplet-based microfluidics and photo-crosslinking, which are then bottom-up assembled via dynamic crosslinking between dopamine-modified hyaluronic acid and phenylboronic acid groups on microgel surface. Results reveal that the microgel assembly can avoid the cell endocytosis of nanoparticles, ensure the high BMSC viability during the regular cell culture, cryopreservation and injection process, promote the chondrogenic differentiation of BMSCs. In addition, animal expriment proves the newborn cartilages present the typical characteristics of articular cartilage. In brief, this microgel assembly not only offers convenience for clinical use (injectability, tissue adhesion) but also provides good microenvironments for chondrogenesis (controlled drug release, interconnected micropores), indicative of its promising application for cartilage repair and regeneration.

In Situ Formation of Microgel Array Via Patterned Electrospun Nanofibers Promotes 3D Cell Culture and Drug Testing in a Microphysiological System.

A microphysiological system (MPS) is recently emerging as a promising alternative to the classical preclinical models, especially animal testing. A key factor for the construction of MPS is to provide a biomimetic three-dimensional (3D) cellular microenvironment. However, it still remains a challenge to introduce extracellular matrix (ECM)-like biomaterials such as hydrogels and nanofibers in a precise and spatiotemporal manner. Herein, we report a strategy to fabricate a MPS combining both electrospun nanofibers and hydrogels. The in situ formation of microsized hydrogel (microgel) array in MPS is realized by patterning electrospun poly(l-lactic acid) (PLLA)/Ca2+ nanofibers via a solvent-loaded agarose stamp and injecting an alginate solution to trigger the quick ionic cross-linking between alginate and Ca2+ released from patterned nanofibers. The one-on-one integration of electrospun nanofibers and microgels not only provides a 3D cellular microenvironment in designated regions in MPS but also improves the stability of these microenvironments under dynamic culture. In addition, due to the biocompatible properties of an ionic cross-linking reaction, patterned cell array can be achieved simultaneously during the microgel formation process. A breast cancer model is then built in MPS by coculturing human breast cancer cells and human fibroblasts in microgel array, and its application in drug (cisplatin) testing is evaluated. Our data prove that MPS-MA offers a more precise platform for drug testing to evaluate the drug concentration, duration time, cancer microenvironment, etc, mainly due to its successful construction of the biomimetic 3D cellular microenvironment.

Engineering the cellular mechanical microenvironment to regulate stem cell chondrogenesis: Insights from a microgel model.

Biophysical cues (especially mechanical cues) embedded in cellular microenvironments show a critical impact on stem cell fate. Despite the capability of traditional hydrogels to mimic the feature of extracellular matrix (ECM) and tune their physicochemical properties via diverse approaches, their relatively large size not only induces biased results, but also hinders high-throughput screening and analysis. In this paper, a microgel model is proposed to recapitulate the role of 3D mechanical microenvironment on stem cell behaviors especially chondrogenesis in vitro. The small diameter of microgels brings the high surface area to volume ratio and then the enlarged diffusion area and shortened diffusion distance of soluble molecules, leading to uniform distribution of nutrients and negligible biochemical gradient inside microgels. To construct ECM-like microenvironment with tunable mechanical strength, three gelatin/hyaluronic acid hybrid microgels with low, medium and high crosslinking densities, i.e., Gel-HA(L), Gel-HA(M) and Gel-HA(H), are fabricated in microfluidic devices by Michael addition reaction between thiolated gelatin (Gel-SH) and ethylsulfated hyaluronic acid (HA-VS) with different substitution degrees of vinyl sulfone groups. Our results show that mouse bone marrow mesenchymal stem cell (BMSC) proliferation, distribution and chondrogenesis are all closely dependent on mechanical microenvironments in microgels. Noteworthily, BMSCs show a clear trend of differentiating into hyaline cartilage in Gel-HA(L) and fibrocartilage in Gel-HA(M) and Gel-HA(H). Whole transcriptome RNA sequencing reveals that mechanical microenvironment of microgels affects BMSC differentiation via TGF-ß/Smad signaling pathway, Hippo signaling pathway and Integrin/YAP/TAZ signaling pathway. We believe this microgel model provides a new way to further explore the interaction between cells and 3D microenvironment. STATEMENT OF SIGNIFICANCE: In recent years, hydrogels have been frequently used to construct 3D microenvironment for cells. However, their relatively large size not only brings biased experimental results, but also limits high-throughput screening and analysis. Herein we propose a gelatin/hyaluronic acid microgel model to explore the effects of 3D cellular mechanical microenvironment (biophysical cues) on BMSC behaviors especially chondrogenesis, which can minimize the interference of biochemical gradients. Our results reveal that BMSC differentiation into either hyaline cartilage or fibrocartilage can be regulated via tailoring the mechanical properties of microgels. Whole transcriptome RNA sequencing proves that "TGF-ß/Smad signaling pathway", "Hippo signaling pathway" and "Integrins/YAP/ TAZ signaling pathway" are activated or inhibited in this process.