The Cornea in Normal Condition and in Groenouw’s Macular Dystrophy

193 Pages·1980·14.066 MB·English

by  Professor Jules François, Virgilio Victoria-Troncoso (auth.)

The Cornea in Normal Condition and
in Groenouw's Macular Dystrophy
The Cornea in Normal Condition and
in Groenouw's Macular Dystrophy
Jules Fran~ois
& Virgilio Victoria-Troncoso
Dr. W. Junk bv Publishers The Hague-Baston-London 1980
Ophthahnological Clinic of the University of Ghent, Belgium.
Director: Professor Jules Franyois.
Also published as volume 47 issl,le 2 of Documenta Ophthalmologica and
supplied to its subscribers as part of their subscription.
© Dr. W. Junk bv Publishers 1980
Softcover reprint of the hardcover 1s t edition 1980
No part of this book may be reproduced and/or published in any form, by
print, photoprint or any other means without written permission from the
publishers.
ISBN-13: 978-94-009-9179-8 e-ISBN-13: 978-94-009-9177-4
001: 10.1007/978-94-009-9177-4
CONTENTS
Introduction . . . . . . 2
Part one: Normal keratocytes 5
Chapter I: Material and methods . . . . . . . . . . .. 7
I. Material 7
II. Methods 8
A. Histological and histochemical techniques. 8
B. Culture techniques for normal keratocytes 9
C. Technique for the study of the corneal architectonic 11
D. Histochemical techniques 12
III. Discussion . . . . . 19
A. Mucopolysaccharides 19
B. Collagen and reticulin 29
C. Lysosomes. 30
D. Mitochondria 34
N. Microscopy . 34
V. Photography. . 36
Chapter II: Normal keratocytes in situ. 37
I. Introduction. . . . 37
II. Personal results. . . 37
Discussion and conclusions 41
Chapter III: Architectonic of the cornea 45
I. Introduction. . 45
II. Personal results. . . 45
A. Flat sections . . 45
B. Transversal sections 48
C. Study of flat sections at the polarisation microscope 48
D. Study of the transversal sections at the polarisation micro-
scope. . . . . . . . . . 50
E. Study of the mucopolysaccharides 50
F. Topo-optical study . . . . . 52
V
G. Diffraction by X-rays 52
H. Examination of collagen-mucopolysaccharide complexes at 53
the high-resolution electron micoscope.
1. Macromolecular model of the corneal stroma 53
Discussion . 54
Chapter IV: The keratocytes in the process of cicatrisation of the
corneal stroma . 61
I. Personal observations. 61
A. Transformation of the keratocytes in a wound 61
B. Histochemical evolution of the keratocytes in the course of
cicatrisation 65
Discussion and conclusions 72
Chapter V: Keratocytes in tissue culture 79
I. Introduction. 79
II. Personal observations. 81
A. Examination of the primary cultures in vivo during growth 81
B. Examination of the ,ub-cultures in vivo during growth 82
C. Examination at t Ie phase-contrast microscope of living
monolayers, mounted in Ringer's solution 82
III. Histochemical study 84
IV. Electron microscopy . 88
V. Scanning microscopy . 89
Discussion and conclusions 90
Part two: Pathological keratocytes (Macular dystrophy of the cornea) 93
Chapter VI . 95
I. Definition of macular dystrophy of the cornea 95
II. Clinical characteristics. 95
III. Heredity 96
Conclusions 97
Chapter VII: Microscopical and histochemical study of macular
dystrophy. 99
Personal observations 99
A. Phase-contrast micr')scopy of fresh sections and of sections
fixed before and after dehydration . 99
B. Optical microscopy of histochemically stained sections 102
C. Polarisation microscopy and to po-optical staining 107
D. Dark-field microscopy 109
Discussions and conclusions 110
VI
Chapter VIII: Transmission electron microscopy of the corneal stroma
in macular dystrophy of the cornea . 113
Personal observations 113
A. Superficial layers of the corneal stroma 113
B. Keratocytes 113
I. First stage of the keratocyte degeneration . 113
II. Second stage of the keratocyte degeneration 114
III. Third stage of the keratocyte degeneration. 118
IV. Fourth and final stage of the keratocyte degeneration 119
C. Intercellular corneal stroma 119
Discussions and conclusions . 120
Chapter IX: Scanning electron microscopy of the corneal stroma in
macular dystrophy. 123
Personal observations 123
Conclusion . 125
Chapter X: Changes of the epithelium, the base membrane, the
endothelium and the Descemet's membrane in macular dystrophy
of the cornea. 127
Personal observations 127
I. Deterioration of the epithelium and the base membrane 127
A. Optical microscopy and histochemistry 127
a. Examination at the clear-field microscope of stained sec-
tions . 127
b. Phase-contrast microscopy of fresh or histochemically
stained sections 129
c. Polarisation microscopy of histochemically stained sec-
tions . 129
d. Dark-field microscopy of stained sections. 130
B. Electron microscopy. 130
C. Scanning electron microscopy 133
II. Changes of the endothelium and the Descemet's membrane 134
Discussion . 137
Conclusion . 138
Chapter XI: Histochemical and microscopical study of corneal cul-
tures from macular dystrophy 139
Personal observations 139
I. Development of the cultures 139
II. Examination of the implantation specimen. 140
III. Microscopy . 140
A. Implanted fragment . 140
VII
Discussion . 149
Conclusions 150
Chapter XII: Transmission electron microscopy of cultures of the
corneal stroma in macular dystrophy. 151
Personal observations 151
Discussion . 155
Conclusions 155
Chapter XIII: Scanning electron microscopy of cultures of macular
dystrophy of the cornea . 157
Personal observations 157
Comparative study of normal keratocytes and keratocytes affected by
macular dystrophy. 157
Conclusions 159
Chapter XN: Vital staining of the lysosomes in normal keratocytes
and in keratocytes from macular dystrophy 161
Introduction . 161
Personal observations 161
I. Normal keratocytes 163
Cell division . 165
Dynamics of the vital staining 166
II. Keratocytcs in macular dystrophy. 167
Dynamics of the vital staining in keratocytes from macular dys·
trophy. 170
Conclusions 170
Chapter XV: Active biological substance present in nonnal kerato·
cytes and capable of acting on the stored mucopolysaccharides
in macular dystrophy of the cornea 173
Personal observations 173
I. Experiment. 173
II. Results. 173
Discussion and conclusions 175
Chapter XVI: Pathogenesis of macular dystrophy of the cornea 177
Chapter XVII: Treatment of macular dystrophy of the cornea 183
Summary 185
Bibliography 188
Subject index 197
VIII
ACKNOWLEDGEMENT
We would like to thank very wannly Dr. Enrique Malbran, who gave us some
corneas affected by macular dystrophy and the facilities of his foundation.
We also would like to thank very sincerely Mrs. Antoinette Victoria-Ihler, who
prepared the tissue cultures, Mr. Andre Uvijls, who made the drawings, Mrs.
Agnes Van Gerven, who prepared the electron·microscopical sections, and
Mrs. Fanny Dhaenens, who typed the manuscript.
1
INTRODUCTION
The three most striking characteristics of the cornea are:
a) Its structure or rather its perfectly regular architectonic, by virtue of
which it is transparent.
b) The absence of vessels, the cornea being nourished by the perilimbic
vessels, the endothelial surface in communication with the aqueous humour
and the epithelial surface in contact with the pre-corneal film.
c) The very slow turnover of the cells, that is to say the keratocytes, with
the result that the metabolism of the cornea is very weak.
It is this third characteristic which justifies our present investigation.
The keratocytes, which are apparently inactive, have in fact a latent
activity. They can be activated by central corneal incisions and also by
tissue cultures. Under either of those conditions, the keratocytes become
very active, develop all the cytoplasmic organites and produce mucopoly
saccharides as well as the precursors of the collagen (Fig. 1).
In order to study the pathological keratocyte, we chose a storage disease,
wherein the catabolism of the mucopolysaccharides is blocked, namely the
macular dystrophy of the cornea.
We undertook the same investigation both for normal and for pathologi
cal corneas and studied the keratocyte 'in situ' and in tissue cultures using
various microscopical and histochemical techniques. In macular dystrophy,
we investigated also the deteriorations secondary to the changes in the
keratocytes.
In view of the fact that, in macular dystrophy, the abnormality occurs at
the level of the lysosomes, we undertook a comparative study 'in vitro' of
the lysosomes of the keratocytes in tissue culture, obtained from normal
corneas and from corneas affected by the disease.
Finally, we will discuss the pathogenesis and the treatment of macular
dystrophy of the cornea.
We have not investigated the role of the keratocytes in inflammatory
conditions of the cornea, which have already been fully developed by nu
merous authors.
2
granules }
by tissue
vacuoles
culture
acid phosphatase
activation
rIffJ
m.
keratocyte "in situ" s.r.e.4'!
I. @ (j)o
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Fig. 1. Activation of the keratocyte by corneal wound or by tissue culture.
3