Cornea - ARVO

ARVO 2013 Annual Meeting Abstracts by Scientific Section/Group - Corneaanalyzed with image processing software. Epithelial migration rate(EMR, in µm/h) and estimated time of healing (ETH, in hours) werecalculated.Results: Density and length of subbasal nerves decreasedsignificantly after 1 month in dry eyes compared to control eyes.Subbasal nerves appeared less branched and the number of epithelialnerve terminals was significantly reduced. These effects were moreprominent 2 months after induction of eye dryness. 4-8 months aftertearing deficiency, density and length of subbasal corneal nerves hadrecovered values close to control, although nerve architecture was notfully normal. EMR was significantly decreased and ETH wassignificantly increased 1 and 6 months after surgery (Table 1),although at month 6th, ETH was partly recovered.Conclusions: The changes in corneal subbasal nerve architecture 1-2months after lacrimal gland removal suggest that nerve damagedevelops shortly after induction of reduced tearing, leading to aneurotrophic slowdown of epithelial wound healing. At longer times,regeneration of corneal nerves appears to restore in part the woundhealing capabilities of the normal corneal epithelium.Commercial Relationships: Kamila Mizerska, None; NicolasCuenca, Universidad de Alicante (P); Carolina Luna, None; SusanaQuirce, None; Laura Fernandez-Sanchez, None; Illes Kovacs,None; M Carmen Acosta, None; Carlos Belmonte, None; JuanaGallar, NoneSupport: SAF2011-22500, CSD2007-00023, IPT-2011-1110-900000 and BFU2008-04425, and in part by BFU2012-36845 andRETICS RD12/0034/0010 (Ministerio de Ciencia e Innovación,Spain, and FEDER, EU)Program Number: 4313 Poster Board Number: C0051Presentation Time: 8:30 AM - 10:15 AMIsolation and characterization of progenitor cells from intactrabbit lacrimal glandHong He 1 , Guoying Sun 1 , Hui Lin 1 , Marie A. Shatos 2 , Darlene A.Dartt 2 , Samuel C. Yiu 1, 3 . 1 Wilmer Eye Institute, Baltimore, MD;2 Shepens Eye Research Institute, Boston, MA; 3 King Khaled EyeSpecialist Hospital, Riyadh, Saudi Arabia.Purpose: Lacrimal gland dysfunction is believed to be the singlemost important factor resulting in aqueous-deficient dry eye.Regeneration of the lacrimal gland using progenitor cells maypotentially restore lacrimal gland function and therefore improve theocular surface health. Previous studies have documented the role ofprogenitor cells in the repair of murine lacrimal glands. Since rabbitis a promising animal model for further lacrimal gland study (due tobetter accessibility to its ocular surface), it is important to studywhether the progenitor cells also exist in rabbit LGs. The purpose ofthis study is to investigate the presence of progenitor cells in intactadult rabbit LGs, and if they could be isolated and expand in vitro.Methods: Frozen intact rabbit LG sections were made. While acinarand duct cells were identified by their respective cellularmorphology, myoepithelial cells were identified byimmunohistochemistry (IHC) using antibody against alpha-smoothmuscle actin (α-SMA). The presence of progenitor cells wasdetermined using IHC by exploring for the expression of selectedstem cell markers - △Np63, ABCG2, Pax6, vimentin. Immature cellswere isolated from LGs using enzyme digestion and mechanicalseparation and subsequently grown in keratinocyte growth medium(KGM) without fetal bovine serum. The expression of selected stemcell markers and α-SMA in immature cells from passage 0 (P0) topassage 2 (P2) was examined by immunocytochemistry (ICC).Results: In the intact adult rabbit LGs, some myoepithlial and ductcells expressed stem cell markers - △Np63, ABCG2, Pax6 andvimentin. Isolated immature cells were found to successfully passageand expand in serum-free KGM, and express stem cell markers -△Np63, ABCG2, Pax6 and vimentin - in a subpopulation from P0 toP2. In addition, some isolated immature cells expressing stem cellmarkers were also found to be labeled with α-SMA.Conclusions: We conclude that progenitor cells exist in intact adultrabbit LGs and can be isolated in vitro. Our results also indicate thatthe progenitor cells may not only be myoepithelial cells but also ductcells.Commercial Relationships: Hong He, None; Guoying Sun, None;Hui Lin, None; Marie A. Shatos, None; Darlene A. Dartt, None;Samuel C. Yiu, NoneProgram Number: 4314 Poster Board Number: C0052Presentation Time: 8:30 AM - 10:15 AMAlterations of Tear Functions and Ocular Surface EpithelialDifferentiation in the SOD-1 Knock- out MouseMURAT DOGRU 1, 2 , Takashi Kojima 2, 1 , Taeko Nagata 2, 1 , AyakoIgarashi 1, 2 , Kazunari Higa 1, 2 , Yoshiyuki Satake 1 , Seika Shimazaki 1 ,Shimizu Takahiko 3 , Kazuo Tsubota 2, 1 , Jun Shimazaki 1 .1 Ophthalmology, Tokyo Dental College, Ichikawa, Japan;2 Ophthalmology, Keio University School of Medicine, Tokyo, Japan;3 Institute of Aging, Chiba University School of Medicine, Chiba,Japan.Purpose: Purpose: SOD-1 knock out mouse has been reported to be amodel for age related dry eye disease. We investigated the alterationsin the tear function and conjunctival ocular surface epithelialdifferentiation in the Sod1-/- in comparison to the wild type mice.Methods: Methods: Ten eyes of 5 Sod1-/- male mice withC57BL/background and 10 eyes of 5 C57BL6 strain wild-type malemice were examined at 10 and 50 weeks in this study. Tear filmstability and corneal epithelial damage was evaluated by fluoresceinand Rose Bengal stainings. Anterior segment photography wascarried out at 10 to 50 weeks. Aqueous tear quantity was measuredwith phenol-red-impregnated cotton threads without anesthesia.Animals were sacrificed and the whole globe specimens underwentPAS and SPDEF(SAM pointed domain containing ets transcriptionfactor) immunohistichemistry staining. Quantitative Real Time-PCRfor conjunctival muc 5AC mRNA and SPDEF expression was alsoperformed. All studies were performed in accordance with the ARVOStatement for the Use of Animals in Ophthalmic and VisionResearch. Statistical analysis was performed by using t test andANOVA. A p value