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Aspiration-assisted intra-operative 3D bioprinting of scaffold-free modular units for skeletal muscle tissue regeneration

ABSTRACT: Traumatic muscle injury associated with volumetric muscle loss results in reversible muscle regeneration and function. Bottlenecks to conventional tissue engineering strategies to augment muscle regeneration are limited by low cell transplantation capacity by conventional injection of dissociated cells, the lack of customization of engineered tissues based on muscle defect geometry, as well as the feasibility of intra-operative use. Here, we developed a facile approach to engineer modular units of scaffold-free high-density muscle tissues in customizable geometric shapes and sizes with integration potential. For the first time, we describe aspiration-assisted intra-operative bioprinting of scaffold-free modular units for preclinical treatment of volumetric muscle loss. We demonstrated the ability of 3D modular muscle units to be bioprinted to the site of the muscle defect, leading to improved muscle, vascular, and neuromuscular regeneration in a mouse model of volumetric muscle loss. Transcriptional profiling revealed that the scaffold-free muscle units enable pre-formed cell-cell interactions that support myogenesis, unlike dissociated cells that lack cell-cell interactions. This technology of intraoperative modular muscle unit implantation is promising for clinical use for precise bioprinting of scalable tissues from geometrically tunable modular units.

ORGANISM(S): Mus musculus Homo sapiens

PROVIDER: GSE292577 | GEO | 2025/05/31

REPOSITORIES: GEO

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Project description:Background & Aims: 3D Bioprinting of an endocrine pancreas is a promising future curative treatment for selected patients with insulin secretion deficiency. In this study we present an end-to-end integrative, scalable concept extending from the molecular to the macroscopic level. Methods: A hybrid scaffold device was manufactured by 3D (bio)printing. INS-1 cells with/without endothelial cells were bioprinted in gelatin methacrylate blend hydrogel. Polycaprolactone was 3D-printed and heparin-functionalized as structural scaffold component. In vitro evaluation was performed by viability and growth assays, total mRNA sequencing, and glucose-stimulated insulin secretion. In vivo, xenotransplantation to fertilized chicken eggs was used to investigate vascularization and function, and finite element analysis modeling served to detect boundary conditions and applicability for human islets of Langerhans. Results: Insulin-secreting pseudoislets were formed and resulted in a viable and proliferative experimental model. Transcriptomics revealed upregulation of proliferative and ß-cell-specific signaling cascades, downregulation of apoptotic pathways, and overexpression of extracellular matrix proteins and VEGF induced by pseudoislet formation and 3D culture. Co-culture with human endothelial cells created a natural cellular niche resulting in enhanced insulin response after glucose stimulation. Survival and function of the pseudoislets after explantation and extensive scaffold vascularization of both the hydrogel and heparinized polycaprolactone components were demonstrated in ovo. Computer simulations of oxygen, glucose, and insulin flows were used to evaluate scaffold architectures and Langerhans islets at a future transplantation site along neurovascular structures. Conclusion: A defined end-to-end process for multidisciplinary bioconvergence research on a bioartificial endocrine pancreas was developed. A modular, patient-specific device architecture is proposed for future research studies.

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Aspiration-assisted intra-operative 3D bioprinting of scaffold-free modular units for skeletal muscle tissue regeneration

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