A C T A O P H T H A L M O L O G I C A VOL. 5 6 1 9 7 8
Department of Ophthalmology (Head: Torstein 1. Bertelsen), School of Medicine, University of Bcrgen, Norway
THE ULTRASTRUCTURE OF THE HUMAN LENS CAPSULE
I. CATARACTOUS LENSES FROM EYES WITH SIMPLE GLAUCOMA A Transmission Electron Microscopic Study BY
JOHAN H. SELAND
The lens capsules from five cataractous lenses removed from eyes with glaucoma simplex have been compared with capsules from five cataractous lenses removed from eyes without glaucoma. Glaucoma simplex, cataracta senilis or the treatment of the glaucoma by drug therapy or surgical intervention do not seem to influence the ultrastructure of the lens capsule. The findings are identical with those observed in the normal aging human lens.
K e y words: lens capsule
-
glaucoma simplex
-
cataracta senilis
-
ultra-
structure.
I t is an old observation that acutely elevated intraocular tension in angle closure glaucoma may give rise to partially reversible opacities of the lens (“Glaukomflecken”; Vogt 1930). These opacities a r e reported to be situated subcapsularly a n d their remnants have been observed to sink down into the cortex as time passes. T h e tissue trauma caused by an acute elevation of the intraocular tension is supposed to be the cause of these changes (Sugar 1957). In recent years some reports of lens changes in simple glaucoma have been published. Higher weight of the lenses from such eyes have been demonstrated (Nakabayashi et al. 1974). Others h a r e shown that the distance between the anterior capsule of the lens a n d the first disjunction zone is increased in glaucoma patients treated with carbonic anhydrase inhibitors a n d reduced in glauReceived April 10, 1978. 715
Johan H . Seland
Age when lens extracted
Pat. No.
Age when glaucoma diagnosed
Period of glaucomatous drug treatment
Time and type of antiglaucomatous operations
Cataractous senilis with glaucoma simpIex: 1. A. J.
8
45
44
2. O.K.
8
52
39
Pilocarpine Epinephrine Acetazolamide
(13 years) (I3 years) ( 3 years)
3. E. S.
8
73
64
Mintachol Pilocarpine Epinephrine Acetazolamide Physiostigmine
( ( ( ( (
4. H.A.
9
74
70
Pilocarpine
( 4 years)
5 . R. S.
9
83
83
Pilocarpine Epinephrine Acetazolamide
( 2 months) ( 2 months) ( 2 months)
0
Trabeculectomy 16 months before extraction
2 years) 8 years) 8 years) 8 years) 3 years) Iridencleisis 4 years before extraction
Cataractous senilis: 6. I.L.
9
7. J.N. 8. S.B. 9. T . P . 10. I. J.
8 8
9 8
69 71 71 73 85
coma patients treated with irreversible choline esterase inhibitors or adrenaline derivatives (Niesel & Bachmann 1974). It has also been shown that the decrease in this lenticular zone preceed t h e development of cataract (Brown & Tripathi 1974).
Capsular Ultrastructure in Glaucoma Simplex
Other links between lens pathology and open angle glaucoma have been established, but here the glaucomas are considered to be of a secondary type. In phacolytic glaucoma a protein component of the lens tissue is thought to gain access to the anterior chamber through a defective lens capsule and thus initiate an inflammatory reaction causing the glaucoma (Flocks et al. 1955). In lenses from eyes with fibrillopathia epitheliocapsularis, the so-called senile exfoliation of the lens capsule, it has been shown that specific changes in the lens capsule occur both in eyes with glaucoma capsulare and those without glaucoma (Bertelsen et al. 1964). As a part of an investigation of the lenticular changes in fibrillopathia epitheliocapsularis it has been found necessary to try and establish if any of the observed lens capsular changes in this disease might be an effect of increased intraocular tension or the treatment there of. This investigation therefore comprises lenses from eyes with glaucoma simplex which have been subjected both to drug therapy and to surgery and comparing these with a group of cataractous lenses removed from eyes without glaucoma.
Material and Methods Ten cataractous lenses extracted from eyes without the use of a-chymotrypsin have been examined. Five of the lenses were extracted from eyes with primary open angle glaucoma. Two of those eyes had undergone surgery and four had
ANT. POLE
Fig. 1 . Subdivision of lens capsule into sections or areas.
717
Johan If. Selund
been treated with a variety of antiglaucomatous drugs (Table I). The remaining five cataractous lenses were extracted from eyes with normal intraocular tension and were used as controls. The lenses were transferred to cold 3 O/O glutaraldehyde in phosphate buffer (pH 7.2) immediately after the operation. Following primary fixation for at least 24 h, the lenses were transsected and the lens nuclei removed. Only the superficial cortex and the capsule were processed further. T h e two halves o f the lenses were postfixed for 2 h in 1 O/O osmium tetraoxide, dehydrated through increasing concentrations of ethanol and propylenoxide before embedding in Epon 8 12. After polymerisation, radial slices were cut and these were again sectioned into eight subsections or areas according to a method described previously (Seland 1974) (Fig. 1). Each subsection was reorientated and reembedded in Epon 812, cut in a Reichert or
Fig. 2
Capsulo-epithelial junction in glaucoma. Note the striking laminations in the lens capsule (LC). Small conglomerates of intertwined osmiophilic fibrils (F) are seen. Epithelial cell (E). Intercellular lacuna (L). x 40 000. Bar: 0.1 urn. Area 4. Pat. No. 1. 718
Capsular Ultrastructiire in Glaucoma Siinplex
LKB ultramicrotome. The sections were stained for 6 min with 1 O/O uranyl acetate and 15 min with a 4 O/O lead citrate and examined in a Philips EM 300 electron microscope. Results Capsular thickness measurements have not been included in this investigation as the study includes lenses of greatly varying ages where the thicknesses vary accordingly. Four of the lens capsules from eyes with glaucoma simplex showed a homogeneous capsular structure a t the anterior pole (Area 1 ) . In one of the lens capsules (lens No. 1) a distinct lamination could be found in this anterior
B
A
Fig. 3. Capsulo-epithelial junction in senile cataract. A. Conspicuous capsular inclusion of the granular type (GI). Normal epithelial cells (E). Lens capsule (LC). x 36 000. Bar: 0.1 ttm. Area 4. Pat. No. 6 . B. Epithelio-capsular junction with fusiform lacunae. Note inclusion bodies (arrows) adhering to epithelium (E) in one lacuna and to the capsule (LC) in the other. x 40 000. Bar: 0.1 pm. Area 3. Pat. No. 7 .
719
Johan H . Seland
central area. Towards the periphery (Area 3 and 4) various degrees of surface parallel lamination of the capsular structure could be observed in all five lens capsules. In between these laminations clefts containing granular inclusions were present in all lenses. The amount and size of these inclusions differed, however, from lens to lens, but their long axes were always surface parallel. Both laminations and intracapsular inclusions were also found in Area 5 (behind the anatomical equator). T h e lens with a laminated capsule in the anterior pole (No. 1) also showed changes a t the epitheliocapsular border. A fibrillar material could be observed (Fig. 2). Groups of intertwined fibrillar material were seen in epithelial lacunae adjacent to the lense capsuIe. T h e thickness of the fibril seemed to be in the region of 25 nm. T h e length of each individual fibril could not be determined. T h e posterior capsules (Areas 6, 7 and 8) all had a homogeneous structure in the five lenses from glaucomatous eyes. Neither laminations nor inclusions could be observed.
Fig. 4. Senile cataract. Granular masses (GM) in a loose matrix adjacent to the lens capsule (LC). Epithelial cell (E). x 40 000. Bar: 0.1 rum. Area 1. Pat. No. 7.
720
Capsular Ultrastructure in Glaucoma Simplex
In the five control lenses removed from eyes with senile cataract and normal intraocular pressure, the lens capsule from the anterior pole all had a homogeneous structure. Towards the periphery (Areas 3, 4 and 5) the phenomenon of intracapsular granular inclusions found in the lenses of glaucomatous eyes were also present in the control lenses. One could not observe any difference in amount, size or structure compared with the lenses from the glaucomatous eyes (with one exception). One of the control lenses not only exhibit the ordinary intracapsular inclusions wedged between the lamination, but also had a striking accumulation of granular intracapsular inclusion with long axis perpendicular to the epithelial surface (Fig. 3A). This inclusion measured at least 1 pm in length and 0.3 p m in width. I t consisted of numerous small dots with a diameter of about 15 nm. Not all dots were circular, but they also seemed to have an eliptical or even a more elongated form. No form of cross-striation could be observed. One of the lenses (No. 7) with a clinical cataract also affecting the central part of the anterior cortex showed accumulation of subcapsular granular material without affecting the ultrastructure of the overlying lens capsule (Fig. 4). At the epithelial capsular junction, small roundish amorphous structures could also be observed situated in spindle-shaped lacunae. These structures could also be seen adhering to the inner surface of the lens capsule when it happened to be stripped off from the epithelial cell. (Fig. 3B). The posterior capsules were all homogeneous without any lamitations or inclusions.
Discussion The lenses removed from glaucomatous eye have all been subjected to some form of pathological stimuli in the form of increased intraocular pressure, application of a variety of potent antiglaucomatous drugs, or to operative intervention. The phenomenon of capsular lamination and its transformation into a homogeneous structure in the anterior polar region have been described in normal lenses as an aging phenomenon (Seland 1974). The one lens which did not adhere to this common pattern, in that it also exhibited laminations in the anterior central area, was the youngest lens where cataract developed two years after operative intervention. One might suspect this opacification to be a type of secondary cataract rather than the naturally developing senile variety. An investigation on this type of cataractous lens seem to support this (Seland 1978). T h e other lens from an eye subjected to operative intervention (lens No. 4) did not exhibit any specific changes in addition to those already described. 721
Johan H . Seland
Treatment of glaucoma with antiglaucomatous drugs does not seem to have any direct effect on the morphology of the lens capsule. The appearance of glaucoma, cataracta senilis, or institution of drugs or surgical therapy for the former condition, do not seem to influence the ultarstructure of the lense capsule.
References Bertelsen T . I., Drablm P. A. & Flood P. R. (1964) T h e socalled senile exfoliation (pseudoexfoliation) of the anterior lens capsu!e, a product of the lens epithelium. Fibrillopathia epitheliocapsularis. Acta ophthal. (Kbh.) 42, 1096-1 113. Brown N. & Tripathi R. (1974) T h e loss of the anterior sub-capsular clear zone of the lens. Prognostic significance in cataract formation. Trans. ophthal. Soc. U . K . 94, 29-45. Flocks M., Littwin C. S. & Zimmermann L. E. (1955) Phacolytic glaucoma. Arch. Ophthal. (Chicago) 54, 37-45. Nakabayashi M., Takatsuki R. & Yoshimura T. (1974) T h e correlation of the lens weight to glaucoma in senile cataractous eyes. d c t a Soc. Ophthal. jab. 78, 1004-1009. Niesel P. & Bachmann E. (1974) Beobachtungen a m Abspaltungsstreifen der Linse bei Glaukomkranken. Albrecht v . Giaefes Arch. klin. exp. Ophthal. 189, 211-217. Seland J. H. (1974) Ultrastructural changes in the normal human lens capsule from birth to old age. Acta ofihthal. (Kbh.) 52, 688-706. Seland J. H . (1978) T h e ultrastructure of the human lens capsule. 11. Cataracta complicata. A transmission electron microscopic study. Acta ophthal. (Kbh.) 56, 723-734. Sugar H. S. (1957) The Glazicomas, p. 216-218. Hoeber & Harper, New York. Vogt A. (1 930) Cataracta disseminata subcapsularis glaucomatosa. Klin. Mbl. Augenheilk. 85, 586-58 7.
Author’s address: Johan H. Seland, Department of Ophthalmology, 5016 Haukeland Sykehus, Norway.
722
Department of Ophthalmology (Head: Torstein 1. Bertelsen), School of Medicine, University of Bcrgen, Norway
THE ULTRASTRUCTURE OF THE HUMAN LENS CAPSULE
I. CATARACTOUS LENSES FROM EYES WITH SIMPLE GLAUCOMA A Transmission Electron Microscopic Study BY
JOHAN H. SELAND
The lens capsules from five cataractous lenses removed from eyes with glaucoma simplex have been compared with capsules from five cataractous lenses removed from eyes without glaucoma. Glaucoma simplex, cataracta senilis or the treatment of the glaucoma by drug therapy or surgical intervention do not seem to influence the ultrastructure of the lens capsule. The findings are identical with those observed in the normal aging human lens.
K e y words: lens capsule
-
glaucoma simplex
-
cataracta senilis
-
ultra-
structure.
I t is an old observation that acutely elevated intraocular tension in angle closure glaucoma may give rise to partially reversible opacities of the lens (“Glaukomflecken”; Vogt 1930). These opacities a r e reported to be situated subcapsularly a n d their remnants have been observed to sink down into the cortex as time passes. T h e tissue trauma caused by an acute elevation of the intraocular tension is supposed to be the cause of these changes (Sugar 1957). In recent years some reports of lens changes in simple glaucoma have been published. Higher weight of the lenses from such eyes have been demonstrated (Nakabayashi et al. 1974). Others h a r e shown that the distance between the anterior capsule of the lens a n d the first disjunction zone is increased in glaucoma patients treated with carbonic anhydrase inhibitors a n d reduced in glauReceived April 10, 1978. 715
Johan H . Seland
Age when lens extracted
Pat. No.
Age when glaucoma diagnosed
Period of glaucomatous drug treatment
Time and type of antiglaucomatous operations
Cataractous senilis with glaucoma simpIex: 1. A. J.
8
45
44
2. O.K.
8
52
39
Pilocarpine Epinephrine Acetazolamide
(13 years) (I3 years) ( 3 years)
3. E. S.
8
73
64
Mintachol Pilocarpine Epinephrine Acetazolamide Physiostigmine
( ( ( ( (
4. H.A.
9
74
70
Pilocarpine
( 4 years)
5 . R. S.
9
83
83
Pilocarpine Epinephrine Acetazolamide
( 2 months) ( 2 months) ( 2 months)
0
Trabeculectomy 16 months before extraction
2 years) 8 years) 8 years) 8 years) 3 years) Iridencleisis 4 years before extraction
Cataractous senilis: 6. I.L.
9
7. J.N. 8. S.B. 9. T . P . 10. I. J.
8 8
9 8
69 71 71 73 85
coma patients treated with irreversible choline esterase inhibitors or adrenaline derivatives (Niesel & Bachmann 1974). It has also been shown that the decrease in this lenticular zone preceed t h e development of cataract (Brown & Tripathi 1974).
Capsular Ultrastructure in Glaucoma Simplex
Other links between lens pathology and open angle glaucoma have been established, but here the glaucomas are considered to be of a secondary type. In phacolytic glaucoma a protein component of the lens tissue is thought to gain access to the anterior chamber through a defective lens capsule and thus initiate an inflammatory reaction causing the glaucoma (Flocks et al. 1955). In lenses from eyes with fibrillopathia epitheliocapsularis, the so-called senile exfoliation of the lens capsule, it has been shown that specific changes in the lens capsule occur both in eyes with glaucoma capsulare and those without glaucoma (Bertelsen et al. 1964). As a part of an investigation of the lenticular changes in fibrillopathia epitheliocapsularis it has been found necessary to try and establish if any of the observed lens capsular changes in this disease might be an effect of increased intraocular tension or the treatment there of. This investigation therefore comprises lenses from eyes with glaucoma simplex which have been subjected both to drug therapy and to surgery and comparing these with a group of cataractous lenses removed from eyes without glaucoma.
Material and Methods Ten cataractous lenses extracted from eyes without the use of a-chymotrypsin have been examined. Five of the lenses were extracted from eyes with primary open angle glaucoma. Two of those eyes had undergone surgery and four had
ANT. POLE
Fig. 1 . Subdivision of lens capsule into sections or areas.
717
Johan If. Selund
been treated with a variety of antiglaucomatous drugs (Table I). The remaining five cataractous lenses were extracted from eyes with normal intraocular tension and were used as controls. The lenses were transferred to cold 3 O/O glutaraldehyde in phosphate buffer (pH 7.2) immediately after the operation. Following primary fixation for at least 24 h, the lenses were transsected and the lens nuclei removed. Only the superficial cortex and the capsule were processed further. T h e two halves o f the lenses were postfixed for 2 h in 1 O/O osmium tetraoxide, dehydrated through increasing concentrations of ethanol and propylenoxide before embedding in Epon 8 12. After polymerisation, radial slices were cut and these were again sectioned into eight subsections or areas according to a method described previously (Seland 1974) (Fig. 1). Each subsection was reorientated and reembedded in Epon 812, cut in a Reichert or
Fig. 2
Capsulo-epithelial junction in glaucoma. Note the striking laminations in the lens capsule (LC). Small conglomerates of intertwined osmiophilic fibrils (F) are seen. Epithelial cell (E). Intercellular lacuna (L). x 40 000. Bar: 0.1 urn. Area 4. Pat. No. 1. 718
Capsular Ultrastructiire in Glaucoma Siinplex
LKB ultramicrotome. The sections were stained for 6 min with 1 O/O uranyl acetate and 15 min with a 4 O/O lead citrate and examined in a Philips EM 300 electron microscope. Results Capsular thickness measurements have not been included in this investigation as the study includes lenses of greatly varying ages where the thicknesses vary accordingly. Four of the lens capsules from eyes with glaucoma simplex showed a homogeneous capsular structure a t the anterior pole (Area 1 ) . In one of the lens capsules (lens No. 1) a distinct lamination could be found in this anterior
B
A
Fig. 3. Capsulo-epithelial junction in senile cataract. A. Conspicuous capsular inclusion of the granular type (GI). Normal epithelial cells (E). Lens capsule (LC). x 36 000. Bar: 0.1 ttm. Area 4. Pat. No. 6 . B. Epithelio-capsular junction with fusiform lacunae. Note inclusion bodies (arrows) adhering to epithelium (E) in one lacuna and to the capsule (LC) in the other. x 40 000. Bar: 0.1 pm. Area 3. Pat. No. 7 .
719
Johan H . Seland
central area. Towards the periphery (Area 3 and 4) various degrees of surface parallel lamination of the capsular structure could be observed in all five lens capsules. In between these laminations clefts containing granular inclusions were present in all lenses. The amount and size of these inclusions differed, however, from lens to lens, but their long axes were always surface parallel. Both laminations and intracapsular inclusions were also found in Area 5 (behind the anatomical equator). T h e lens with a laminated capsule in the anterior pole (No. 1) also showed changes a t the epitheliocapsular border. A fibrillar material could be observed (Fig. 2). Groups of intertwined fibrillar material were seen in epithelial lacunae adjacent to the lense capsuIe. T h e thickness of the fibril seemed to be in the region of 25 nm. T h e length of each individual fibril could not be determined. T h e posterior capsules (Areas 6, 7 and 8) all had a homogeneous structure in the five lenses from glaucomatous eyes. Neither laminations nor inclusions could be observed.
Fig. 4. Senile cataract. Granular masses (GM) in a loose matrix adjacent to the lens capsule (LC). Epithelial cell (E). x 40 000. Bar: 0.1 rum. Area 1. Pat. No. 7.
720
Capsular Ultrastructure in Glaucoma Simplex
In the five control lenses removed from eyes with senile cataract and normal intraocular pressure, the lens capsule from the anterior pole all had a homogeneous structure. Towards the periphery (Areas 3, 4 and 5) the phenomenon of intracapsular granular inclusions found in the lenses of glaucomatous eyes were also present in the control lenses. One could not observe any difference in amount, size or structure compared with the lenses from the glaucomatous eyes (with one exception). One of the control lenses not only exhibit the ordinary intracapsular inclusions wedged between the lamination, but also had a striking accumulation of granular intracapsular inclusion with long axis perpendicular to the epithelial surface (Fig. 3A). This inclusion measured at least 1 pm in length and 0.3 p m in width. I t consisted of numerous small dots with a diameter of about 15 nm. Not all dots were circular, but they also seemed to have an eliptical or even a more elongated form. No form of cross-striation could be observed. One of the lenses (No. 7) with a clinical cataract also affecting the central part of the anterior cortex showed accumulation of subcapsular granular material without affecting the ultrastructure of the overlying lens capsule (Fig. 4). At the epithelial capsular junction, small roundish amorphous structures could also be observed situated in spindle-shaped lacunae. These structures could also be seen adhering to the inner surface of the lens capsule when it happened to be stripped off from the epithelial cell. (Fig. 3B). The posterior capsules were all homogeneous without any lamitations or inclusions.
Discussion The lenses removed from glaucomatous eye have all been subjected to some form of pathological stimuli in the form of increased intraocular pressure, application of a variety of potent antiglaucomatous drugs, or to operative intervention. The phenomenon of capsular lamination and its transformation into a homogeneous structure in the anterior polar region have been described in normal lenses as an aging phenomenon (Seland 1974). The one lens which did not adhere to this common pattern, in that it also exhibited laminations in the anterior central area, was the youngest lens where cataract developed two years after operative intervention. One might suspect this opacification to be a type of secondary cataract rather than the naturally developing senile variety. An investigation on this type of cataractous lens seem to support this (Seland 1978). T h e other lens from an eye subjected to operative intervention (lens No. 4) did not exhibit any specific changes in addition to those already described. 721
Johan H . Seland
Treatment of glaucoma with antiglaucomatous drugs does not seem to have any direct effect on the morphology of the lens capsule. The appearance of glaucoma, cataracta senilis, or institution of drugs or surgical therapy for the former condition, do not seem to influence the ultarstructure of the lense capsule.
References Bertelsen T . I., Drablm P. A. & Flood P. R. (1964) T h e socalled senile exfoliation (pseudoexfoliation) of the anterior lens capsu!e, a product of the lens epithelium. Fibrillopathia epitheliocapsularis. Acta ophthal. (Kbh.) 42, 1096-1 113. Brown N. & Tripathi R. (1974) T h e loss of the anterior sub-capsular clear zone of the lens. Prognostic significance in cataract formation. Trans. ophthal. Soc. U . K . 94, 29-45. Flocks M., Littwin C. S. & Zimmermann L. E. (1955) Phacolytic glaucoma. Arch. Ophthal. (Chicago) 54, 37-45. Nakabayashi M., Takatsuki R. & Yoshimura T. (1974) T h e correlation of the lens weight to glaucoma in senile cataractous eyes. d c t a Soc. Ophthal. jab. 78, 1004-1009. Niesel P. & Bachmann E. (1974) Beobachtungen a m Abspaltungsstreifen der Linse bei Glaukomkranken. Albrecht v . Giaefes Arch. klin. exp. Ophthal. 189, 211-217. Seland J. H. (1974) Ultrastructural changes in the normal human lens capsule from birth to old age. Acta ofihthal. (Kbh.) 52, 688-706. Seland J. H . (1978) T h e ultrastructure of the human lens capsule. 11. Cataracta complicata. A transmission electron microscopic study. Acta ophthal. (Kbh.) 56, 723-734. Sugar H. S. (1957) The Glazicomas, p. 216-218. Hoeber & Harper, New York. Vogt A. (1 930) Cataracta disseminata subcapsularis glaucomatosa. Klin. Mbl. Augenheilk. 85, 586-58 7.
Author’s address: Johan H. Seland, Department of Ophthalmology, 5016 Haukeland Sykehus, Norway.
722
