A bioprosthetic ovary created using 3D printed microporous scaffolds restores ovarian function in sterilized mice

The advancement of tissue and, ultimately, organ engineering requires the ability to pattern human tissues composed of cells, extracellular matrix, and vasculature with controlled microenvironments that can be sustained over prolonged time periods. To date, bioprinting methods have yielded thin tissues that only survive for short durations. To improve their physiological relevance, we report a method for bioprinting 3D cell-laden, vascularized tissues that exceed 1 cm in thickness and can be perfused on chip for long time periods (>6 wk). Specifically, we integrate parenchyma, stroma, and endothelium into a single thick tissue by coprinting multiple inks composed of human mesenchymal stem cells (hMSCs) and human neonatal dermal fibroblasts (hNDFs) within a customized extracellular matrix alongside embedded vasculature, which is subsequently lined with human umbilical vein endothelial cells (HUVECs). These thick vascularized tissues are actively perfused with growth factors to differentiate hMSCs toward an osteogenic lineage in situ. This longitudinal study of emergent biological phenomena in complex microenvironments represents a foundational step in human tissue generation.

Supramolecular hydrogels are used in the 3D printing of high-resolution, multi-material structures. The non-covalent bonds allow the extrusion of the inks into support gels to directly write structures continuously in 3D space. This material system supports the patterning of multiple inks, cells, and void spaces.

Gelatin is a commonly used natural polymer which is derived from collagen. The isoelectric point of gelatin can be modified during the fabrication process to yield either a negatively charged acidic gelatin, or a positively charged basic gelatin at physiological pH. This theoretically allows electrostatic interactions to take place between a charged biomolecule and gelatin of the opposite charge, forming polyion complexes. Various forms of gelatin carrier matrices can be fabricated for controlled-release studies, and characterization studies have been performed which show that gelatin carriers are able to sorb charged biomolecules such as proteins and plasmid DNA through polyion complexation. The crosslinking density of gelatin hydrogels has been shown to affect their degradation rate in vivo, and the rate of biomolecule release from gelatin carriers has been shown to have a similar profile, suggesting that complexed gelatin/biomolecule fragments are released by enzymatic degradation of the carrier in vivo. This review will emphasize how biomolecules released from gelatin controlled-release systems are able to retain their biological activity, allowing for their use in tissue engineering, therapeutic angiogenesis, gene therapy, and drug delivery applications.