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Guoying Sun, Jennifer H. Elisseeff, Peter J. Mcdonnel, Samuel C. Yiu; Decellularized Rabbit Lacrimal Gland As A Scaffold For Engineering An Artificial Lacrimal Gland. Invest. Ophthalmol. Vis. Sci. 2012;53(14):636.
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Dry eye is a chronic condition caused by a decline in tears secretion. Patients with nonfunctioning lacrimal glands or blocked ducts, such as those with Stevens-Johnson syndrome, chemical and thermal injuries, or ocular cicatricial pemphigoid, have very limited options because of the short duration and action of lubricating agents. A bioartificial lacrimal gland will greatly benefit many of these patients. This study is focused on new methods for artificial lacrimal gland tissue engineering.
NZW rabbit lacrimal glands extracted from orbits were treated with 1% SDS or 1% triton X-100 at RT or 4°C for 36 hours, followed by DNase(2000u/ml) 2 hours at 37°C and 0.3% peracetic acid 3 hours treatment, washed with PBS after each treatment, stored in PBS with 1% strep/pen antibiotics at 4°C. Histological analysis including H&E, IHC, DAPI stain and DNA analysis were performed to evaluate the efficiency of decellularization and to determine the ECM components that remained after processing. To evaluate biological activity, freshly isolated lacrimal gland acinar cells were seeded onto decellularized lacrimal scaffolds and constructs were cultured over 28 days.
Lacrimal gland treatment with 1% SDS for 36 hours effectively removed the cells and treatment at 4°C best preserved the original shape and mechanical integrity of the lacrimal glands. Furthermore, 1% SDS was more efficient in removing cellular components and DNA compared to 1% triton X-100 under similar conditions. The decellularized lacrimal gland had the form of a three dimensional fibrous scaffold network that supported cell attachment. Cells survived on the scaffolds over the 28 days culture and acinar-like structures with a central lumen were formed.
Decellularized tissues are natural scaffolds that provide an ideal mimic of the physical and biological cues of native extra cellular matrix architecture. This study provides the initial steps in developing a new three-dimensional scaffold for artificial lacrimal gland tissue engineering.
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