Waffle-inspired hydrogel-based macrodevice for spatially controlled distribution of encapsulated therapeutic microtissues and pro-angiogenic endothelial cells.

Macro-encapsulation systems for delivery of cellular therapeutics in diabetes treatment offer major advantages such as device retrievability and high cell packing density. However, microtissue aggregation and absence of vasculature have been implicated in the inadequate transfer of nutrients and oxygen to the transplanted cellular grafts. Herein, we develop a hydrogel-based macrodevice to encapsulate therapeutic microtissues positioned in homogeneous spatial distribution to mitigate their aggregation while concurrently supporting an organized intra-device network of vascular-inductive cells. Termed Waffle-inspired Interlocking Macro-encapsulation (WIM) device, this platform comprises two modules with complementary topography features that fit together in a lock-and-key configuration. The waffle-inspired grid-like micropattern of the "lock" component effectively entraps insulin-secreting microtissues in controlled locations while the interlocking design places them in a co-planar spatial arrangement with close proximity to vascular-inductive cells. The WIM device co-laden with INS-1E microtissues and human umbilical vascular endothelial cells (HUVECs) maintains desirable cellular viability in vitro with the encapsulated microtissues retaining their glucose-responsive insulin secretion while embedded HUVECs express pro-angiogenic markers. Furthermore, a subcutaneously implanted alginate-coated WIM device encapsulating primary rat islets achieves blood glucose control for 2 weeks in chemically induced diabetic mice. Overall, this macrodevice design lays foundation for a cell delivery platform, which has the potential to facilitate nutrients and oxygen transport to therapeutic grafts and thereby might lead to improved disease management outcome.

[Comparative study on Sichuan yak pericardium and Australian cattle pericardium].

Currently, as the key raw material of artificial biological heart valve, bovine pericardium is mainly depend on import and has become a "bottleneck" challenge, greatly limiting the development of domestic biological heart valve. Therefore, the localization of bovine pericardium is extremely urgent. In this study, the pericardium of Sichuan yak was compared with that of Australian cattle in terms of fundamental properties and anti-calcification performance. The results demonstrated that the appearance and thickness of yak pericardium were more advantageous than the Australian one. Sichuan yak pericardium and Australian cattle pericardium had comparable performance in shrinkage temperature, mechanical test and anti-calcification test. This study preliminarily verifies the feasibility of substitution of Australian cattle pericardium by Sichuan yak pericardium and promotes the progression of bovine pericardium localization with data support.

Degradation regulated bioactive hydrogel as the bioink with desirable moldability for microfluidic biofabrication.

Bioink development is vital in biofabriacation for generating three-dimensional (3D) tissue-like constructs. As potential candidates of bioinks, hydrogels need to meet the requirements of good moldability, initially strong mechanical properties and prominent bioactivity to guarantee cell vitality and further assembly. Enzyme-induced dynamic degradation is an efficient and biocompatible approach to improve the bioactivity of hydrogels through releasing space continuously for cell proliferation and promoting the functional establishing of engineered tissue. Here a novel bioink was designed by introducing alginate lyase into composite Alginate-GelMA hydrogels. Results showed that bioink with proper lyase content exhibited desirable modability and cytocompatibility. Then cell-laden osteon-like microfibers were engineered with the microfluidic device and diverse complex 3D constructs were also successfully assembled. This degradation-regulated bioink showed great promise in a variety of applications in tissue engineering and biomedical investigation.

Microfluidic-based generation of functional microfibers for biomimetic complex tissue construction.

UNLABELLED: Microfluidic-based fiber system displays great potential in reconstructing naturally complex tissues. In these systems, fabrication of the basic fiber is a significant factor in ensuring a functional construction. The fiber should possess the strong mechanical rigidity for assembly, predefined microenvironment for cell spatial distribution and high biocompatibility for cell functional expression. Herein we presented a composite material by the combination of methacrylated gelatin (GelMA) and alginate for fiber engineering with capillary microfluidic device. Being regulated by GelMA incorporation, the composite hydrogels exhibited higher mechanical moduli, better stretching performance, and lower swelling compared to pure alginate one. On the basis of the composite material and capillary microfluidic device, we constructed the double-layer hollow microfibers to simulate complex tissues. The microfibers could be precisely controlled in size and multi-layered structure by varying flow rates and outlet diameter, and it showed satisfied application in woven-structure assembly. As an example to mimic a functional tissue, a biomimetic osteon-like structure was fabricated by encapsulating human umbilical vascular endothelial cells (HUVECs) in middle layer to imitate vascular vessel and human osteoblast-like cells (MG63) in the outer layer to act role of bone. During the incubation period, both MG63 and HUVECs exhibited not only a robust growth, but also up-regulated gene expression. These results demonstrated this microfluidic-based composite microfibers system is a promising alternative in complex tissue regeneration. STATEMENT OF SIGNIFICANCE: Cell-laden microfibers based on microfluidic device is attracting interest for reconstructing naturally complex tissues. One shortage is the lack of suitable materials which satisfy microfluidic fabrication and cell biofunctional survival. This study reports the first combination of alginate-GelMA composite and capillary-based microfluidic technology. The composite materials possess high mechanical properties for fabrication and assembly, and tunable environment for cell spatial encapsulation. Significantly, the engineered double-layer hollow microfiber with osteon-like structure showed enhanced cellular bioactivity and realized initially functional establishment. This microfluidic-based composite microfiber not only explores a competitive candidate in complex tissues reconstruction, but also expands the biological application of microfluidic technology. This developing interdisciplinary area should be widely interested to the readers of biofabrication, biomaterials and tissue engineering.

Photo-cross-linkable methacrylated gelatin and hydroxyapatite hybrid hydrogel for modularly engineering biomimetic osteon.

Modular tissue engineering holds great potential in regenerating natural complex tissues by engineering three-dimensional modular scaffolds with predefined geometry and biological characters. In modular tissue-like construction, a scaffold with an appropriate mechanical rigidity for assembling fabrication and high biocompatibility for cell survival is the key to the successful bioconstruction. In this work, a series of composite hydrogels (GH0, GH1, GH2, and GH3) based on a combination of methacrylated gelatin (GelMA) and hydroxyapatite (HA) was exploited to enhance hydrogel mechanical rigidity and promote cell functional expression for osteon biofabrication. These composite hydrogels presented a lower swelling ratio, higher mechanical moduli, and better biocompatibility when compared to the pure GelMA hydrogel. Furthermore, on the basis of the composite hydrogel and photolithograph technology, we successfully constructed an osteon-like concentric double-ring structure in which the inner ring encapsulating human umbilical vascular endothelial cells (HUVECs) was designed to imitate blood vessel tubule while the outer ring encapsulating human osteoblast-like cells (MG63s) acts as part of bone. During the coculture period, MG63s and HUVECs exhibited not only satisfying growth status but also the enhanced genic expression of osteogenesis-related and angiogenesis-related differentiations. These results demonstrate this GelMA-HA composite hydrogel system is promising for modular tissue engineering.

A spatial patternable macroporous hydrogel with cell-affinity domains to enhance cell spreading and differentiation.

Cell adhesion and spreading are two essential factors for anchorage-dependent cells such as osteocytes. An adhesive macroporous hydrogel system, in which cell-affinitive domains and sufficient cytoskeleton reorganization space were simultaneously constructed, was proposed in this report to support cell adhesion and spreading, respectively, and facilitate cell differentiation and function establishment eventually. The adhesive macroporous alginate hydrogel was developed by RGD peptide graft and gelatin microspheres hybridization to generate cellular adhesion sites and highly interconnected macropores. The successful stretched morphology and enhanced osteogenic differentiation of MG-63 cells in this modified alginate hydrogel showed clearly the feasibility that cell function may be effectively facilitated. Besides, this hydrogel model can be further applied to construct complex micropatterned structure, such as individual microgels in shapes of circle, square, cross and ring, and osteon-like structure containing both osteogenic and vascularized area generated by a double-ring assembly. These results should provide this adhesive macroporous photocrosslinkable hydrogel system as potential three-dimensional scaffolds for guiding tissue formation, especially for the bioengineering of tissues that have multiple cell types and require precisely defined cell-cell and cell-substrate interactions.

Bottom-up approach to build osteon-like structure by cell-laden photocrosslinkable hydrogel.

Based on photocrosslinkable PEGDMA and GelMA hydrogels, two "bottom-up" approaches ("circle-and-cross" and "layer-by-layer") were successfully developed to construct osteon-like structures with microchannel networks. Significantly, the "layer-by-layer" approach employing the GelMA hydrogel with a higher biocompatibility was more favorable for building biomimetic osteon.