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Joana Witt, Gerd Geerling, Sonja Mertsch, Stefan Schrader, Kristina Spaniol; Decellularized porcine conjunctiva for conjunctival reconstruction. Invest. Ophthalmol. Vis. Sci. 2017;58(8):4362.
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© ARVO (1962-2015); The Authors (2016-present)
Conjunctival repair is an essential part of ocular surface reconstruction. Due to limitations of currently used substitute tissues such as amniotic membrane (AM), there is a need for the development of new matrices for conjunctival reconstruction. This study explored porcine decellularized conjunctiva (PDC) as an alternative conjunctival substitute in-vitro and in-vivo.
Porcine conjunctiva was decellularized, cell removal confirmed histologically and residual DNA quantified by a fluorescence assay. Preservation of extracellular matrix (ECM) was examined by collagen quantification and electron microscopy. Primary human conjunctival epithelial cells (CEC) were incubated with PDC supernatant or culture medium (control) for cytotoxicity determination. Elongation at break (EB) was measured using a material testing machine (n=4). Conjunctival defects were produced in New Zealand White rabbits and closed using PDC or AM (n=6 per group). Defect size, amount of sutures, conjunctival hyperemia and fornix depth (FD) were measured after surgery and at day 3 and 10. Epithelialization and lymphocytic infiltration were assessed histologically after euthanasia (day 10).
PDC did not contain cellular structures and the DNA content was significantly reduced (p<0.001). The ECM ultrastructure was preserved and the collagen content was preserved (70.23±14.35 µg/mg, p=0.4). PDC revealed no cytotoxic effects on CEC compared to control (p=0.99). EB of PDC was significantly higher compared to AM (19.68±0.96 vs 12.30±0.45 mm, p<0.001). At day 10, 6 of 6 PDC were present compared to 3 of 6 AM and the PDC group exhibited significantly less suture-loss (p<0.001). Conjunctival hyperemia, FD and lymphocytic infiltration revealed mild inflammation and did not differ significantly between PDC and AM at day 3 and 10. Histologically, PDC were well integrated into the conjunctival wound and covered with partially multilayered epithelium after 10 days.
Decellularization of porcine conjunctiva generates a cell free, stable and non-immunogenic matrix, which is well tolerated after xenogeneic transplantation. Compared to AM, PDC reveals enhanced extensibility and stability in-vitro and in-vivo. As porcine decellularized tissues are frequently applied clinically in other fields, PDC might offer an easily available substitute for conjunctival reconstruction. Its use for fornix reconstruction has to be to be elucidated in future studies.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.
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