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Fine Structural Alterations of Corneal Endothelium During Experimental Uveitis

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Fine Structural Alterations of Corneal Endothelium During Experimental Uveitis Fine structural alterations of corneal endothelium during experimental uveitis Hajime Inomata and George K. Smelser Uveitis was produced in albino rabbits by intravitreal injection of bovine serum albumin. By this often-used technique, a uveitis became manifest about 7 days after the antigen injection. Inflammation was most severe in the anterior portion of the uvea but also involved the cornea, which became thickened and hazy. Blood vessels invaded the anterior stroma from the limbus and in advanced cases reached half way to the center of the cornea and the middle, as well as the anterior layers of the stroma. Marked haze teas noted in the anterior chamber, and evidence of cells and fibrin clots could be seen by slit lamp examina- tion. Structural changes of the endothelium included mononuclear cell infiltration and vacuolization. The cells were lymphocytes and plasma cells. Although polymorphonuclear leukocytes were observed in the anterior chamber, none was found invading the endothelium. The actual invasion itself consisted of the cells entering between endothelial cells, so that they came to lie first in the inter endothelial cell space and finally under the endothelium, between it and Descemet's membrane. During this process the endothelial cell cytoplasm showed little evidence of change. Their cytoplasmic organelles were well preserved, but there was a remarkable change in the dilation of the intercellular space. The functional complexes which characterized the anterior chamber aspect of these cells appeared intact, even though lymphocytes and plasma cells lay under them and between endothelial cells, and apparently had passed through these junctions. The changes which have been described were more pronounced in the inferior aspect of the cornea where massive keratic precipitates occurred. In no instance did inflammatory cells invade Descemet's membrane. It is concluded, therefore, that they were derived entirely from the anterior chamber, which they had presumably entered from the iris of ciliary processes. Key words: allergic uveitis, experimental production, corneal endothelium, corneal stroma, Descemet's membrane, corneal vascularization, corneal infiltrate, corneal inflammations, histopathology, ultrastructure, rabbits. From the Department of Ophthalmology, Columbia .he histopathology of experimental University, New York, N. Y. uveitis has been studied1"11 since zur Nid- This investigation was supported, in part, by den first demonstrated that an ocular in- National Institutes of Health Research Grant No. 8 RO-1EY-00190-13, from the National Eye flammation was elicited when a foreign Institute, and, in part by Public Health Service blood serum was injected into the vitreous Research Career Program Award No. 5-K6-NB- body of a rabbit eye. Corneal involvement 19-609-06 from the National Institute of Neuro- was observed in addition to the hyper- logical Diseases and Stroke, and by a Fight for sensitivity reaction of uveal tissue, and it Sight Post-Doctoral Research Fellowship No. was shown that corneal opacification occurs F-210 (G-2) financed by Fight for Sight, New 12 York City. in eyes with uveitis. Keratic precipitates, Manuscript submitted Nov. 21, 1969; manuscript which are found by slit lamp examination accepted Dec. 29, 1969. in the clinic as deposition of precipitates 272 Downloaded from iovs.arvojournals.org on 10/01/2021 Volume 9 Alterations of corneal endothelium during uveitis 273 Number 4 on corneal endothelium, are an important Azure II, were made for light microscopy. Thin sign of uveitis. sections were cut with glass and diamond knives 5 7 on a Porter-Blum ultramicrotome and mounted Very few histological studies " have on Parlodion-coated copper mesh, doubly stained been made on the corneal changes ac- with uranyl acetate and lead citrate, and examined companying experimental uveitis, and in a Siemens Elmiskap 1. these have been confined to light micros- Observations copy. The present study is concerned with the histopathology of the corneal endo- Light microscopy thelium in experimental uveitis at the level Early stage. Although the major part of the electron microscope, and with com- of the cornea still remained clear, the parison to that which is known in clinical periphery of the inferior quadrant was conditions. vascularized and turbid, and the aqueous humor contained a small quantity of pre- Materials and methods cipitates. Several sections from different An immune uveitis was produced by injection parts of the cornea were made. of 0.1 ml. of 20 per cent sterile aqueous bovine Neither edematous changes nor cell serum albumin solution* into the vitreous humor infiltration was detected in any part of of adult albino rabbits. After 1 to 2 weeks, during which the cornea, as well as the rest of the eye, the epithelium. Vascularization of the sub- was free of visible symptoms, pericorneal vascular epithelial layer of the stroma was accom- injection appeared. The aqueous humor became panied by slight edematous swelling and turbid and contained cells, as seen by slit lamp cell infiltration in the periphery of the examination. The cornea became vascularized, lateral and inferior quadrants of the cor- and precipitates were noted on its posterior sur- face. When inflammation was fully advanced, the nea. Descemet's membrane was completely cornea became extremely turbid and the anterior free of inflammatory cells. The endothelium chamber contained masses of cells, a hypopyon. in the inferior quadrant, where corneal The iris was severely hyperemic, and the animals vascularization and turbidity were most were photophobic. Corneal opacity and aqueous pronounced, was irregular in thickness, humor turbidity became less pronounced when the uveitis subsided. Corneas in several stages due to the infiltration of many inflamma- of this inflammation were prepared for electron tory cells. A small amount of cellular microscopy. infiltration in the endothelium was also Rabbits were anesthesized by intravenous injec- seen in the lateral, but was absent in the tion of sodium pentobarbital solution,! 12 mg. superior quadrant. Precipitates in the per pound of body weight. Immediately after enucleation a coronal incision was made at the anterior chamber consisted of a small equator of the globe and the anterior half of number of mononuclear cells such as plas- the eye was immersed in fixative. Under observa- ma cells, lymphocytes, and monocytes, tion with a binocular dissecting microscope, the within a thick network of fibrin. Some lens and iris were gently removed while the cells were found close to the endothelial anterior segment was still immersed in the fixative. The cornea was cut into several large sections surface as keratic precipitates in the in- and the sclera was removed with sharp razor ferior quadrant (Fig. 1). blades. Great care was taken to prevent mechan- Advanced stage. A few days later, more ical damage to the endothelial surface. The severe responses were recognizable (Fig. fixative was 1 per cent osmium tetroxide buffered 2). The cornea became extremely turbid with 0.15M sodium cacodylate, pH 7.4, at 4° C, in which the tissue was kept for one hour, then and the anterior chamber contained a mas- dehydrated rapidly through a graded series of sive hypopyon. In spite of the extreme alcohols. The large corneal segments were trimmed turbidity of the cornea as a whole, the into pieces 1 by 2 mm. in 80 per cent alcohol, epithelium remained free from edema. and embedded in Epon 812 after treatment in Considerable changes, however, were seen propylene oxide. Thick sections, stained with in the stroma and the endothelium. Newly formed blood vessels in the anterior cor- "Nutritional Biochemical Corp., Chargin Falls, Ohio. fNemhutal, Abbott Laboratories, North Chicago, 111. neal stroma reached halfway from the Downloaded from iovs.arvojournals.org on 10/01/2021 274 Inomata and Smelser Investigative Ophthalmology April 1970 rig. J.. renpnerai (jurnea at eariy stage 01 lnuammauuii. lniiainmaiory etuis are aireauy seen on the endothelial surface as keratic precipitates before stromal change becomes manifest. See also Fig. 6. Ep, Epithelium; V, vessel; St, stroma; AC, anterior chamber. (x200.) Fig. 2. Peripheral cornea in advanced stage showing pronounced neovascularization and cellular infiltration of the entire cornea. Vacuolization is seen in the endothelial cell layer. Ep, Epithelium; V, vessel; St, stroma; AC, anterior chamber. (xl30) Fig. 3. Epithelium (Ep) shows little change through all stages of the inflammation; only small edematous vacuoles (arrows) and a few cell infiltrations in the basal portion, in advanced stage. S£, stroma; V, vessel. (x400.) Downloaded from iovs.arvojournals.org on 10/01/2021 Volume 9 Alterations of corneal endothelium during uveitis 275 Number 4 limbus to the center and were also present filtration were limited to the periphery in the middle layers of the periphery, par- and no epithelial edema was detected. ticularly in the inferior quadrant. Edem- The endothelium seemed to be almost atous swelling was very great in the intact. The central, superior quadrant of anterior third and posterior third layers the cornea was free from cellular infil- of the stroma and slightly less in the middle tration. layer. Mononuclear cells were scattered Electron microscopy. The fine structural through almost the entire stroma, much observations were concentrated on the en- more around the newly formed blood dothelium. Since the structure of inflam- vessels than in the other regions. Most matory
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