Abstract
Purpose :
Piezoelectric drop-on-demand inkjet 3d Bio-printing was developed to support various liquid substances, providing precise control over material deposition. The production of bioinks and printing protocols for this technology is under development and might be applyed to several laboratory and medical conditions, that require precise cell or othe biometaerial deposition. The purpose of this study was to test the feasibility of filling a digital image-based cornea wounds by bioinks.
Methods :
Tissuejet 1000 (MicroFab Technologies, TX, USA) piezzoeletric dispenser was used to produce water based bioinks. Drop-on-demand bursts and continuous ejection were both used to achieve an optimal droplet regarding to size, drop velocity and trajectory.
- Bioink viability
Corneal epithelial cells (CEC) were obtained from corneoescleral rims, cultured in SHEM medium and expanded in vitro. CEC in suspension were poured through to the bioprint to a culture plate. Morphology and migration were evaluated by phase contrast microscopy and its viability analysed by MTT method.
- Biopatch printing protocol
Images of different types of human cornea lesions were obtained and digitally manipulated. A image compound of a white shape, with the same lesion shape, in a black background were created. This image was saved as monochrome bitmap to create the print script file to be applyed onto the corneal surface.
Results :
Epithelial cell viability was determined by MTT analysis and cells integrity (OU CONFLEINCIA) were monitored using phase contrast microscopy for three days during its growth at 37 °C. Image manipulation of human corneal wound were possible to be obtained and digitally inverted to be inputed in the 3D bioprinter software. The script provided the cells and biomaterials to be accurately positioned in established patterns for printing corneal biopatches.
Conclusions :
The development of printing protocols and the feasibility analysis of bioinks are tools that make viable the Piezoelectric Inkjet Printing technique for the manufacturing of corneal bio-patches. The protocol and bioink resulting from this work are being used for diverse customized precise microvolume biological applications by our group. This work is currently in progress and initial results have been promising.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.