The aim of this study was to test the surgical handling and clinical applicability of a three-dimensional (3D) printed scaffold fabricated from gelatin (FDA approved material) for conjunctival reconstruction.

3D printing technique was employed to fabricate scaffolds using a 15% (w/v) solution of gelatin (Type A, from porcine skin, Bioreagent grade). The scaffolds were then post processed by 2 hours cross linking using 12mg/ml carbodiimide (EDC) and 1.5mg/ml N-Hydroxysuccinimide (NHS) to enhance mechanical properties. Two scaffolds with approximate thickness of 300 µm (3 layers) and 400 µm (4 layers) and approximate pore size of 550-650 µm were examined. The surgical handling of each scaffold was tested by three surgeons (not informed about the composition) and sutured 3 times per surgeon and compared with amniotic membrane (AM). In vitro viability, proliferation and adhesion of Human adipose driven stem cells (SC) on scaffolds were evaluated with MTT assay; direct microscopy and scanning electron microscopy (SEM). Each procedure was repeated 3 times.

The scaffolds had favorable optical characteristics (Colorless and transparent), satisfactory tensile strength and the surgical handling was better compared to AM however they were less suturable. SC’s cultivated on scaffolds showed satisfactory viability and proliferation in MTT assay and direct microscopy. In addition to corroborating the observations made by direct microscopy, SEM images showed appropriate cell adhesion to the scaffold and scattered cell masses on the surface of the scaffold.

3D printed gelatin scaffolds can be looked as a promising bio-artificial tissue substitute for conjunctival reconstruction.