IADR Abstract Archives

Magnetic Bioprinting Improves Viability of Three-Dimensional Lacrimal Gland Organoids

Objectives: Establishment of exocrine gland organoids using three-dimensional (3D) bioprinting platforms is necessary to overcome the lack of consistency and clinical translation of current organoid biofabrication protocols using xenogeneic matrices. Magnetic 3D bioprinting (M3DB) is a novel bioprinting technology successful used by our research group to generate functional salivary gland secretory epithelial organoids. However, there is a lack of bioprinting protocols to develop other exocrine organs like lacrimal glands (LG) organoid, which is essential to discover new drugs for dry eye disease (DED). This study aimed to develop scalable, consistent, xenogeneic-free, and scaffold-free culture system for LG organoids.
Methods: Primary cells from porcine LG were cultured in serum-free expansion media in ultra-low attachment plates. The viability and spheroid formation efficiency were determined until day 9. Secretory acinar and proliferative markers were identified including Aquaporin 5, Cytokeratin 14, E-Cadherin, and Ki67 by immunocytochemistry and qPCR. Next, LG cells were tagged with magnetic nanoparticles and allowed to assemble under the same condition as non-magnetized cells. Cell viability, spheroid formation efficiency and differentiation markers were statistically compared.
Results: Non-magnetized LG cells formed a spheroid-like cell aggregate (lacrisphere) after 48h. The lacrisphere number and size significantly increased until day 6 and 9, respectively. These outcomes were correlated with Ki67 expression. At day 9, branching morphogenesis and lumen formation in lacrispheres were observed; moreover, Aquaporin 5, Cytokeratin 14 and E-Cadherin have strong immunofluorescence. The use of M3DB platform shortened lacrisphere formation to 24h. M3DB-derived lacrispheres showed a consistent lacrisphere size and significant improvement in cell viability when compared with non-magnetized LG cells.
Conclusions: M3DB technology offers a feasible platform for a consistent LG organoid biofabrication in a porcine proof-of-concept model. Different muscarinic agonists will be tested to validate the functionality of this organoid to further investigate LG physiology, pathogenesis, and therapeutic strategies for DED.

2021 South East Asian Division Meeting (Hong Kong)
Hong Kong
2021
069
Salivary Research
  • Rodboon, Teerapat  ( Chulalongkorn University , Pathum Wan , Thailand )
  • Yodmuang, Supansa  ( Chulalongkorn University , Pathum Wan , Thailand )
  • Chaisuparat, Risa  ( Chulalongkorn University , Bangkok , Bangkok , Thailand )
  • Ferreira, Joao  ( Chulalongkorn University Faculty of Dentistry , Bangkok , Thailand )
  • NONE
    National Research Council of Thailand (NRCT), mid-career research grant number NRCT5-RSA63001-12
    Oral Session
    Dental materials and biomaterials III
    Wednesday, 12/08/2021 , 03:15PM - 04:45PM