Evaluation of Decellularized Porcine Jejunum as a Matrix for Lacrimal Gland Reconstruction In Vitro for Treatment of Dry Eye Syndrome.

Purpose Dry eye syndrome (DES) can cause blindness in severe cases, but mainly palliative treatments exist. A tissue-engineered lacrimal gland (LG) could provide a curative treatment. We aimed to evaluate decellularized porcine jejunum (SIS-Muc) as a scaffold for porcine LG epithelial cells. Methods To evaluate SIS-Muc as a potential scaffold, basement membrane proteins in SIS-Muc and native LG were compared (immunohistochemistry [IHC]). Porcine LG epithelial cells cultured on plastic were characterized (immunocytochemistry), and their culture supernatant was compared with porcine tears (proteomics). Epithelial cells were then seeded onto SIS-Muc in either a static (cell crown) or dynamic culture (within a perfusion chamber) and metabolic (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) and secretory capacities (β-hexosaminidase assay), protein expression (IHC), and ultrastructure transmission electron microscopy (TEM) compared in each. Results Collagen IV and laminin were found in both native LG and SIS-Muc. When cultured on plastic, LG epithelial cells expressed pan-cytokeratin, Rab3D, HexA, and produced mucins, but lysozyme and lactoferrin expression was nearly absent. Some porcine tear proteins (lipocalin-2 and lactoferrin) were found in LG epithelial cell culture supernatants. When LG cells were cultured on SIS-Muc, metabolic and β-hexosaminidase activities were greater in dynamic cultures than static cultures (P < 0.05). In both static and dynamic cultures, cells expressed pan-cytokeratin, Rab3D, lysozyme, and lactoferrin and produced mucins, and TEM revealed cell polarization at the apical surface and cell-cell and cell-scaffold contacts. Conclusions SIS-Muc is a suitable scaffold for LG cell expansion and may be useful toward reconstruction of LG tissue to provide a curative treatment for DES. Dynamic culture enhances cell metabolic and functional activities.

[1]  K. Dou,et al.  Pig Liver Xenotransplantation: A Review of Progress Toward the Clinic , 2016, Transplantation.

[2]  S. Schrader,et al.  Development of Causative Treatment Strategies for Lacrimal Gland Insufficiency by Tissue Engineering and Cell Therapy. Part 2: Reconstruction of Lacrimal Gland Tissue: What Has Been Achieved So Far and What Are the Remaining Challenges? , 2016, Current eye research.

[3]  Heike Walles,et al.  A Combined 3D Tissue Engineered In Vitro/In Silico Lung Tumor Model for Predicting Drug Effectiveness in Specific Mutational Backgrounds. , 2016, Journal of visualized experiments : JoVE.

[4]  J. Elisseeff,et al.  Three-Dimensional Culture of Functional Adult Rabbit Lacrimal Gland Epithelial Cells on Decellularized Scaffold. , 2016, Tissue engineering. Part A.

[5]  P. Lazarovici,et al.  Bioengineering the Lung : Molecules , Materials , Matrix , Morphology , and Mechanics Revascularization of decellularized lung scaffolds : principles and progress , 2015 .

[6]  S. Aicher,et al.  Denervation of the Lacrimal Gland Leads to Corneal Hypoalgesia in a Novel Rat Model of Aqueous Dry Eye Disease. , 2015, Investigative ophthalmology & visual science.

[7]  B. Greve,et al.  Engineering of a Secretory Active Three-Dimensional Lacrimal Gland Construct on the Basis of Decellularized Lacrimal Gland Tissue. , 2015, Tissue engineering. Part A.

[8]  Antonia W Godehardt,et al.  Review on porcine endogenous retrovirus detection assays—impact on quality and safety of xenotransplants , 2015, Xenotransplantation.

[9]  R. Gangnus,et al.  An engineered 3D human airway mucosa model based on an SIS scaffold. , 2014, Biomaterials.

[10]  E. Akpek,et al.  Dry Eye: an Inflammatory Ocular Disease , 2014, Journal of ophthalmic & vision research.

[11]  W. Wurst,et al.  High-fat diet induced isoform changes of the Parkinson's disease protein DJ-1. , 2014, Journal of proteome research.

[12]  Eben Alsberg,et al.  Decellularized tissue and cell-derived extracellular matrices as scaffolds for orthopaedic tissue engineering. , 2014, Biotechnology advances.

[13]  J. Hornegger,et al.  Morphological Features of the Porcine Lacrimal Gland and Its Compatibility for Human Lacrimal Gland Xenografting , 2013, PloS one.

[14]  M. Nardi,et al.  Biological parameters determining the clinical outcome of autologous cultures of limbal stem cells. , 2013, Regenerative medicine.

[15]  H. Walles,et al.  Tissue Engineering of a Human 3D in vitro Tumor Test System , 2013, Journal of visualized experiments : JoVE.

[16]  Guang-Yan Yu,et al.  Microvascular autologous transplantation of partial submandibular gland for severe keratoconjunctivitis sicca , 2013, British Journal of Ophthalmology.

[17]  E. Campos,et al.  Diagnostic performance of a tear protein panel in early dry eye , 2013, Molecular vision.

[18]  F. Paulsen,et al.  Schirmer strip vs. capillary tube method: non-invasive methods of obtaining proteins from tear fluid. , 2013, Annals of anatomy = Anatomischer Anzeiger : official organ of the Anatomische Gesellschaft.

[19]  H. Walles,et al.  Upcyte® microvascular endothelial cells repopulate decellularized scaffold. , 2013, Tissue engineering. Part C, Methods.

[20]  M. Ali,et al.  Establishing Human Lacrimal Gland Cultures with Secretory Function , 2012, PloS one.

[21]  Harald C Ott,et al.  Organ engineering based on decellularized matrix scaffolds. , 2011, Trends in molecular medicine.

[22]  J. V. van Best,et al.  Long-term follow-up after submandibular gland transplantation in severe dry eyes secondary to cicatrizing conjunctivitis. , 2010, American journal of ophthalmology.

[23]  Jan Hansmann,et al.  Vascularised human tissue models: a new approach for the refinement of biomedical research. , 2010, Journal of biotechnology.

[24]  S. Schrader,et al.  Cultivation of lacrimal gland acinar cells in a microgravity environment , 2009, British Journal of Ophthalmology.

[25]  M. Trousdale,et al.  Microporous poly(L-lactic acid) membranes fabricated by polyethylene glycol solvent-cast/particulate leaching technique. , 2009, Tissue engineering. Part C, Methods.

[26]  Jan Hansmann,et al.  Engineered liver-like tissue on a capillarized matrix for applied research. , 2007, Tissue engineering.

[27]  T. Wedel,et al.  Amniotic membrane as a carrier for lacrimal gland acinar cells , 2007, Graefe's Archive for Clinical and Experimental Ophthalmology.

[28]  Ronald E. Smith,et al.  Transepithelial bioelectrical properties of rabbit acinar cell monolayers on polyester membrane scaffolds. , 2007, American journal of physiology. Cell physiology.

[29]  Carolyn G. Begley,et al.  The epidemiology of dry eye disease: report of the Epidemiology Subcommittee of the International Dry Eye WorkShop (2007). , 2007, The ocular surface.

[30]  Ronald E. Smith,et al.  Tissue-engineered tear secretory system: functional lacrimal gland acinar cells cultured on matrix protein-coated substrata. , 2007, Journal of biomedical materials research. Part B, Applied biomaterials.

[31]  Ting Wang,et al.  Polyethersulfone dead-end tube as a scaffold for artificial lacrimal glands in vitro. , 2006, Journal of biomedical materials research. Part B, Applied biomaterials.

[32]  Athanasios Mantalaris,et al.  Proliferation rates of HepG2 cells encapsulated in alginate are increased in a microgravity environment compared with static cultures. , 2005, Artificial organs.

[33]  Linda G Griffith,et al.  Engineering principles of clinical cell-based tissue engineering. , 2004, The Journal of bone and joint surgery. American volume.

[34]  Stephen F Badylak,et al.  The extracellular matrix as a scaffold for tissue reconstruction. , 2002, Seminars in cell & developmental biology.

[35]  C. Framme,et al.  [Transplantation of the submandibular gland in absolute dry eyes. Effect on the ocular surface]. , 2002, Klinische Monatsblatter fur Augenheilkunde.

[36]  Donald C. Chang,et al.  Growth of purified lacrimal acinar cells in Matrigel raft cultures. , 2002, Experimental eye research.

[37]  S. Pflugfelder,et al.  Substrate modulation of morphology, growth, and tear protein production by cultured human lacrimal gland epithelial cells. , 1995, Experimental cell research.

[38]  M. Montgomery,et al.  Immunocytochemical localization of lysozyme and surfactant protein A in rat type II cells and extracellular surfactant forms. , 1992, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.