Figure S1. Schematic of two extrusion-based bioprinting methods. (A,B) The syringe and set of printers on the left are driven pneumatically by using air pressure and the associated syringes lack the graduations necessary for quantifying volumes dispensed. (C) The Hamilton syringe and Organovo bioprinter on the right is driven by direct mechanical force on the plunger. The Hamilton syringe features graduations for exact volume quantification, depicted on the right.
IntroductionOne challenge in tissue engineering (TE) is designing a construct which ensures adequate oxygen diffusion and removal of waste throughout the implanted tissue. In native tissue, the maximum distance between any cell and capillary is limited to the diffusion distance of oxygen, or 100–200 μm1. Mimicking this dense microvasculature in TE constructs is critical to cell viability and function, particularly in thick implants, such as those for skeletal muscle repair2. There have been many recent advancements in using TE to develop skeletal muscle constructs for the treatment of volumetric muscle loss (VML) in injuries incurred by soldiers and civilians. We have developed a tissue engineered muscle repair (TEMR) technology which consists of myoblasts seeded onto a porcine bladder acellular matrix (BAM). While TEMR has previously been reported to result in 60–70% functional recovery in rodent models within 2 months after implantation3,4, we hypothesize that incorporating endothelial cells (ECs) and pericytes in the TEMR will accelerate the rate and magnitude of functional recovery by facilitating anastomosis with the recipient microcirculation and increasing perfusion into the construct.ObjectiveUse bioprinting to position endothelial cells and pericytes with myoblasts on the BAM to “pre‐vascularize” the TEMR construct.ResultsDecellularized microvasculature basement membrane remains intact in some areas of the BAM (Figure 1A). Using the Organovo NovoGen Bioprinter®, we have developed a novel bioprinting method to pattern ECs and myoblasts on the acellular bladder matrix (BAM). ECs seeded onto the BAM begin to form network‐like structures after just 24 hours (Figure 1B). EC viability was shown to be high (>90%) for two bioprinting needle diameters (250 μm vs 500 μm). (Figure 1C).ConclusionsThe presence of ECM from microvasculature on the BAM suggests that the BAM is a promising scaffold for pre‐vascularization. This is further supported by the formation of endothelial network‐like formations on the BAM after just 24 hrs. It is also encouraging that the 250 μm bioprinting needle does not reduce cell viability, as a smaller diameter allows for more precise patterning of cells. Together, these preliminary results suggest that it is feasible to “pre‐vascularize” the TEMR construct by bioprinting.Future WorkWe will further investigate EC viability and network formation in co‐culture with myoblasts. We also plan to incorporate bioprinted pericytes to further encourage and support the formation of pre‐vascularized microvascular networks in TEMR.Support or Funding InformationNIH T32 GM008715, Organovo, Inc., and the UVA Center for Advanced BiomanufacturingThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
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