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Kristina Spaniol, Alexander Kunze, Marco Metzger, Gerd Geerling, Stefan Schrader; Decellularization of porcine lacrimal gland tissue for development of a lacrimal gland scaffold. Invest. Ophthalmol. Vis. Sci. 2013;54(15):914.
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© ARVO (1962-2015); The Authors (2016-present)
In cases of severe dry eye due to lacrimal gland insufficiency engineering of a lacrimal gland tissue construct in vitro might be a promising future treatment approach. Aim of this study was to evaluate structure and main basement membrane components of native porcine lacrimal gland tissue before and after a decellularization process in order to develop an acellular scaffold for lacrimal gland regeneration.
Lacrimal glands were extracted from six domestic pigs after euthanasia. Six biological replicates were used for this study. Each gland was cut into four pieces, two of them were left native and the other two were decellularized using sodium desoxychelate in ultra-pure water over night. Tissue pieces were embedded in paraffin as well as OCT and the morphology of native and decellularized tissue was examined histologically by hematoxylin and eosin (H&E) staining. Additionally, expression of basement membrane markers, such as laminin, collagen IV and fibronectin was evaluated by immunostaining.
Histology showed an intact connective tissue matrix after the decellularization process. Immunohistochemistry revealed the expression of major basement membrane components such as collagen IV and laminin in native lacrimal gland tissue and these components were still detectable after the decellularization of the lacrimal gland tissue. Efficacy of the decellularization process was demonstrated by complete absence of nuclei in the lacrimal gland tissue, as assessed by DAPI-staining.
Decellularization of lacrimal gland tissue generates an intact acellular scaffold with preserved acinar structures, containing major basement membrane components such as collagen IV and laminin. An intact basement membrane is a prerequisite for essential cellular processes like adhesion, migration as well as proliferation and therefore decellularized lacrimal gland tissue might be a promising scaffold for lacrimal gland regeneration in vitro.
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