Purchase this article with an account.
S. Selvam, P.B. Thomas, C.A. Peng, M.D. Trousdale, D. Stevenson, J.E. Schechter, A.K. Mircheff, J.T. Jacob, R.E. Smith, S.C. Yiu; Bioengineering an Artificial Lacrimal Gland: Study of Functional Rabbit Lacrimal Gland Acinar Cells Cultured on Matrix Protein–Coated Substrates . Invest. Ophthalmol. Vis. Sci. 2006;47(13):1931.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
To study the morphological and physiological properties of rabbit lacrimal acinar cells cultured on matrix protein–coated biopolymers, such as silicone, collagen I, and copolymers of poly–D,L–lactide–co–glycolide (PLGA; 85:15 and 50:50) and poly–L–lactic acid (PLLA) for the development of a tissue–engineered tear secretory system.
Purified rabbit lacrimal gland acinar cells were obtained from aseptically excised inferior lacrimal glands. The fabricated biopolymers were sterilized and placed in a 24–well tissue culture plate. The polymeric samples were divided into two study groups; one group was coated with Matrigel®, an extracellular matrix protein and the other group with no Matrigel® coating served as a control. Cell suspensions at a density of 5 x 105/ml were seeded onto the plate. On day 6, sample sets from each group were fixed with Karnovsky fixative for 2 hrs at 4oC and prepared for scanning and transmission electron microscopy (SEM and TEM) studies to evaluate the morphological properties of the cells. To study the physiological properties, two additional sets of samples from the two groups, one stimulated with carbachol and the other unstimulated, were incubated for 30 min and a standard secretion assay was used to evaluate for ß–hexosaminidase activity secreted to the medium.
SEM detected no significant difference in cell morphology between the two study groups for any of the biopolymers. TEM revealed best preservation of the histiotypic characteristics of lacrimal acinar cells (i.e., secretory granules, exocytosis, junctional complexes and microvilli) in cells cultured on Matrigel®–coated PLGA and PLLA, compared with cells cultured on uncoated materials. Collagen promoted ductal epithelial cell development. The TEM study could not be performed on silicone because it was too soft for TEM sectioning. Carbachol stimulation induced a significant increase in ß–hexosaminidase secretory activity in cells cultured on all biopolymers, except collagen.
One of the biopolymers tested, PLLA, showed the greatest promise in maintaining the morphological and physiological properties of purified rabbit lacrimal gland epithelial cells for the fabrication of a tissue–engineered tear secretory system.
This PDF is available to Subscribers Only