CHAPTER 6 

The Vitreous
Henrik Lund-Andersen & Birgit Sander

Introduction tube of primary vitreous surrounded by the secondary vitre-

ous, running from the retrolental space to the optic nerve
The vitreous body makes up approximately 80% of the (area of Martegiani). The tube is called Cloquet’s canal (Fig.
volume of the eye and thus is the largest single structure of 6.1); this is not a liquid-filled canal, but simply a portion of
the eye (Fig. 6.1). In the anterior segment of the eye, it is differentiated gel devoid of collagen fibrils.
delineated by and adjoins the ciliary body, the zonules, and The term tertiary vitreous is related to the fibrillary mate-
the lens. In the posterior segment of the eye, the vitreous rial, which develops as the suspensor fibrils, the zonules, of
body is delineated by and adjoins the retina. the lens. During childhood the vitreous undergoes signifi-
The vitreous body has many normal physiological func- cant growth. The length of the vitreous body in the newborn
tions. This chapter focuses on the most important physio- eye is approximately 10.5 mm, and by the age of 13 years,
logic relationships, especially those that have a close clinical the actual length of the vitreous increases to 16.1 mm in the
correlation. As background for the understanding of the male.107 In the absence of refractive changes, the mean adult
physiology and the pathophysiology of the vitreous body, vitreous is 16.5 mm.30,91
we focus on the main features of the anatomy, biochemistry,
and biophysics. Molecular and cellular considerations
The investigation of the vitreous body and its structure of embryology
and function is hampered by two fundamental difficulties.
The two main components of the vitreous, collagen and
Firstly any attempts to define vitreous morphology are in fact
hyaluronic acid, are produced in the primary and secondary
attempts to visualize a tissue, which by design is intended to
vitreous. In the primary vitreous, however, there is initial
be invisible. Secondly the various techniques that have previ-
production of substances other than hyaluronic acid, such
ously been employed to define the structure of the vitreous
as galactosaminoglycans; later hyaluronic acid becomes the
body are combined with artifacts that make interpretations
predominant constituent.27,41,111,127
difficult in terms of the true in vivo physiological
The primary vitreous contains cells which in the second-
situation.
ary vitreous differentiate as hyalocytes and fibroblasts. The
hyalocytes are believed to be involved in the production of
Anatomy glycosaminoglycans, especially hyaluronic acid, a non-
sulfated glycosaminoglycan.111
Although the function of the fibroblasts is not known
Embryology exactly, they are probably involved in the formation of col-
Structural considerations of embryology lagen. The retina may also be a source of collagen synthe-
sis.88,108 The hyalocytes are found in the vitreous cortex,
In the early stages the optic cup is mainly occupied by the approximately 30 µm from the internal limiting membrane
lens vesicle. As the cup grows the space formed is filled by a (ILM), with the highest density near the vitreous base and
system of fibrillar material, presumably secreted by the cells the posterior pole.9
of the embryonic retina. Later, with the penetration of the
hyaloid artery, more fibrillar material apparently originating
from the cells of the wall of the artery and other vessels
Anatomy of the mature vitreous body
contribute to filling the space. The combined mass is known The mature vitreous body is a transparent gel which occupies
as primary vitreous.22,23,39,110,127 the vitreous cavity. It has an almost spherical appearance,
The secondary vitreous develops later, appearing at the end except for the anterior part, which is concave, corresponding
of the sixth week, and is associated with the increasing size to the presence of the crystallin lens. The vitreous body is a
of the vitreous cavity and the regression of the hyaloid vas- transparent gel; however, it is not completely homogeneous
cular system. The main hyaloid artery remains for some (Fig. 6.2). The outermost part of the vitreous, called the
time, but it eventually disappears and leaves in its place a cortex, is divided into an anterior cortex and a posterior

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 Anatomy

Wieger's Berger's Cloquet's cortex, the latter being approximately 100 µm thick (Fig.
ligament space canal 6.3). The cortex is also called the anterior and the posterior
hyaloid. The cortex consists of densely packed collagen
fibrils (Fig. 6.4). The vitreous base (see Fig. 6.1) is a three-
dimensional zone. It extends approximately from 2 mm
anterior to the ora serrate to 3 mm posterior to the ora
serrata, and it is several millimeters thick. The collagen fibrils
are especially densely packed in this region.
Fovea
The vitreoretinal interface
The vitreoretinal interface can be defined from electron
microscopy as the outer part of the vitreous cortex (posterior
hyaloid), including anchoring fibrils of the vitreous body
and the ILM of the retina (Fig. 6.5).29,34,44–46,93 The ILM is a
retinal structure between 1 and 3 µm thick, consisting
Cortex mainly of type IV collagen and proteoglycans.117 It contains
several layers and can be considered the basal lamina of the
Müller cells, the foot processes of which are in close contact
with the membrane.
Vitreous The vitreous cortex is firmly attached to the ILM in the
base vitreous base region, around the optic disc (Weiss ring), at
the vessels, and in the area surrounding the foveola at a
Figure 6.1  Sketch of the primate eye showing the vitreous body and its diameter of 500 µm.48,106 Under normal conditions, the con-
relations. Wieger’s ligament is the attachment of the vitreous to the lens. nection between the fibrils of vitreous cortex and the ILM is
Berger’s space and the Cloquet’s canal are the former sites of the hyaloid
looser than in the rest of the vitreoretinal interface. The
artery. (From Heegaard 1997.45)
adhesion is strong in young individuals, and dissection of

A B

Figure 6.2  Human vitreous dissection. (A) Vitreous of a 9-month-old child. The sclera, choroid and retina were dissected off the vitreous, which remains
attached to the anterior segment. A band of gray tissue can be seen posterior to the ora serrata. This is peripheral retina that was firmly adherent to the
posterior vitreous base and could not be dissected. The vitreous is solid and although situated on a surgical towel exposed to room air maintains its shape
because at this age the vitreous is almost entirely gel. (B) Human vitreous dissected off the sclera, choroid and retina are still attached to the anterior
segment. The specimen is mounted on a lucite frame using sutures through the limbus and is then immersed in a lucite chamber containing an isotonic,
physiologic solution that maintains the turgescence of the vitreous and avoids collapse and artefactual distortion of vitreous structure. (Reprinted with
permission from Sebag & Balazs 1984.114)

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 SECTION 2  Physiology of optical media   Chapter 6   The Vitreous

the retina from the vitreous often leaves ILM tissue adherent
to the vitreous cortex.47,60,108,109 Under pathologic conditions,
Ultrastructural, biochemical, and
the tight connections between the vitreous cortex and the biophysical aspects
ILM play an important role, as is discussed later in this
chapter. Ultrastructural and biochemical aspects
The vitreous contains more than 99% water; the rest is com-
posed of solids. The vitreous acts as a gel (i.e. an intercon-
nected meshwork) that surrounds and stabilizes a large
amount of water compared with the amount of solids. The
gel structure of the vitreous results from the arrangement of
long, thick, non-branching, collagen fibrils suspended in
a network of hyaluronic acid, which stabilize the gel struc-
ture and the conformation of the collagen fibrils (Figs 6.6
and 6.7).8,10,107
In the human eye the major part of the glycosaminogly-
can is hyaluronic acid, with a molecular weight of 3–4.5 ×
106.111 The volume of non-hydrated hyaluronic acid is
0.66 cm3/g, in contrast with the volume of the hydrated
molecule, which is 2000–3000 cm3/g.8 The molecule forms
into large, open coils, with the anionic sites spread apart.
This arrangement of small-diameter fibers, separated by
highly hydrated glycosaminoglycan chains, permits the
transmission of light to the retina with minimal scattering.11
Figure 6.3  Human vitreous structure during childhood. This view of the
The collagen fibrils in the vitreous are thin, with diameters
central vitreous from an 11-year-old child demonstrates a dense vitreous of approximately 10–20 nm. Collagen fibrils are mostly of
cortex with hyalocytes. The posterior aspect of the lens is seen below, collagen type II. They are composed of three identical
though dimly illuminated. No fibers are present in the vitreous. α-chains, which form a triple helix. The helix is stabilized by
hydrogen bonds between opposing residues in different
chains.111 Collagen type IX is also present and may function
as a bridge, linking type II collagen fibrils together.10,32,120
Collagen V/XI is integrated with collagen II in the collagen
fibers.81 The collagen fibrils seem to interconnect with the
hyaluronic acid, most likely via bridging glucoproteins.111
The viscoelastic properties of the vitreous gel are neither due
to hyaluronic acid or collagen alone but to the combination
of the two molecules.128
Dissolved in the water of the vitreous gel are inorganic
and organic substances as shown in Table 6.1, where plasma
values are given for comparison.
According to Table 6.1, it appears that gradients exist in
both directions between vitreous and plasma.3,79,82,97,98 These
gradients are a result of several mechanisms: presence of the
blood–ocular barriers (i.e. active and passive passage across
the barriers), metabolism in retina and ciliary body, and
diffusion processes in the vitreous body (Box 6.1).
The values in Table 6.1 represent mean values for the
Figure 6.4  Ultrastructure of human vitreous cortex. Scanning electron
whole vitreous. The methods used to quantitate vitreous
microscopy demonstrates the dense packing of collagen fibrils in the
vitreous cortex. To some extent this arrangement is exaggerated by the concentrations are difficult and may differ between studies
dehydration that occurs during specimen preparation for scanning electron in absolute numbers. However, regional differences within
microscopy (magnification = ×3750). the vitreous have been measured for some substances.12

Figure 6.5  Sketch of the vitreoretinal interface /

vitreoretinal border region (VBR). The VBR consists of
Vitreous cortex two major components: the anchoring fibrils of the
vitreous body and membrana limitans interna (MLI).
The MLI is composed of three structures: the fusing
Anchoring part point of the anchoring vitreous fibrils, lamina densa
VBR MLI Lamina densa and lamina lucida. M = Müller cell. (From Heegaard
M M M Lamina lucida 1997.45)

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 Ultrastructural, biochemical, and biophysical aspects

Figures 6.8 to 6.10 show the regional difference for glucose

(Fig. 6.8), lactate (Fig. 6.9), and oxygen (Fig. 6.10). The fall
in vitreous oxygen tension towards the center, correspond-
ing to the upper curve in Figure 6.10, was also found by
Sakaue100 and seems to result from an oxygen flux from
the retina towards the vitreous corresponding to arterioles;
A the flux goes in the opposite direction corresponding to the
venules (lower curve). Several studies have found an increase
in preretinal oxygen after photocoagulation, indicating that
IF the oxygen supply to the inner retina improves after destruc-
tion of the outer retina and a concomitant decrease in tissue
metabolism and oxygen needs.40,85,94,122–124

Hyaluronic acid Collagen fibril Box 6.1  Vitreous – aging and ocular pathology

C • The concentration of salts and organic substances of the

vitreous differ substantially from plasma due to the blood–
aqueous and blood–retinal barrier
• Small molecules move through the vitreous gel by diffusion
• Vitreous fluorometry is useful for evaluation of the vitreal
morphology, the fluorescein profile is an indicator of
physiologic aging such as vitreous liquefaction
• The aging process leads to posterior vitreous detachment,
easily visualized by optical coherence tomography (OCT)
Interconnecting filament • Vitreoretinal traction may lead to formation of a macular hole
Chondroitin sulfate and the traction can be conducted through the retinal layers
• Vitreoretinal traction is also implicated in some cases of
Figure 6.6  Ultrastructure of hyaluronic acid/collagen interaction in the macular edema
vitreous. Specimen was fixed in glutaraldehyde/paraformaldehyde and
stained with ruthenium red. Collagen fibrils (C) are coated with amorphous
• In diabetes, the high glucose speeds up metabolism before
material (A) believed to be hyaluronic acid. The amorphous material may visible retinopathy
connect to the collagen fibril via another glycosaminoglycan, possibly • Increased demand for oxygen and capillary closure leads to
chondroitin sulfate (see inset). Interconnecting filaments (IF) appear to retinal ischemia and an increased production of VEGF
bridge between collagen fibrils, inserting or attaching at sites of hyaluronic • Increased leakage through the blood–retinal barrier leads to
acid adhesion to the collagen fibrils (bar = 0.1 µm). (Reprinted with permission macular edema
from: Asakura A. Histochemistry of hyaluronic acid of the bovine vitreous body as • VEGF inhibition and steroids decrease macular edema
studied by electron microscopy. Acta Soc Ophthalmol J 1985; 89:179.)

Connecting fibril

Liquid
channel

Fiber

A B Na-Hyaluronate
molecular coils

Figure 6.7  Ultrastructure of human vitreous. (A) Specimens were centrifuged to concentrate structural elements, but contained no membranes or
membranous structures. Only collagen fibrils were detected. There were also bundles of parallel collagen fibrils such as the one shown here in cross-section
(arrow). (B) Schematic diagram of vitreous ultrastructure, depicting the dissociation of hyaluronic acid (HA) molecules and collagen fibrils. The fibrils
aggregate into bundles of packed parallel units. The HA molecules fill the spaces between the packed collagen fibrils and form “channels” of liquid vitreous.
(Reprinted with permission from Sebag & Balazs 1989.113 Reproduced with permission from Association for Research in Vision and Ophthalmology.)
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 SECTION 2  Physiology of optical media   Chapter 6   The Vitreous

Table 6.1  Concentration of various substances in the vitreous (weighted averages in mmol/kg H2O)
Inorganic substances
Sodium Potassium Calcium Magnesium Chloride Phosphate pH
Vitreous 134 9.5 5.4* 2.3* 105 2 7.29**
Plasma 143 5.6 9.9* 2.2* 97 0.4 7.41**
Organic substances
Ascorbate Glucose Lactate
Vitreous 0.46 3.0 12.0
Plasma 0.04 5.7 10.3
82
*Human data from McNeil et al 1999.
**Porcine data from Andersen 1991.3
All other values are rabbit data from Reddy & Kinsey 1960;97 with modification from Kinsey 1967.59

60

Glucose
n = 20

Plasma: 3,77
± 0,14
n = 18 45
PO2 (mm Hg)

A
V

I
2,75 2,97 2,92
± 0,23 ± 0,08 ± 0,08 30

Figure 6.8  Glucose concentration in different parts of the vitreous body

and in plasma. All values are in µmol/g tissue weight (mean ± standard
deviation, n = 20). (From Bourwieg et al 1974.12 Reproduced with permission from
Association for Research in Vision and Ophthalmology.)

15
0 250 500 750 1000
x (µm)

Figure 6.10  Heterogeneity of the PO2 in the preretinal vitreous of a

Lactate non-photocoagulated eye. Graphic representation of the PO2 (± SEM)
n = 20 recorded when the O2-sensitive microelectrode was withdrawn from the
vitreal surface of the retina (x = 0) towards the vitreous. Curve A: opposite
an arteriole: curve I: opposite an intervascular zone: curve V: opposite a vein.
Plasma: 4,22
These results are averages of measurements made on one or both eyes of
± 0,47
11 miniature pigs. (From: Mohar I. Effect of laser photocoagulation on oxygenation
n = 18
of the retina in miniature pigs. Invest Ophthalmol Vis Sci 1985; 26:1410. Reproduced
with permission from Association for Research in Vision and Ophthalmology.)

Biophysical aspects
5,07 5,31 5,33 The gel structure acts as a barrier against movement of
± 0,43 ± 0,38 ± 0,47 solutes. Basically, substances may move by two different
processes: diffusion or bulk flow. The diffusion process can
Figure 6.9  Lactate concentration in different parts of the vitreous body
and in plasma. All values are in µmol/g tissue weight (mean ± standard be illustrated in humans by using fluorescein as a tracer
deviation, n = 20). (From Bourwieg et al 1974.12 With kind permission of Springer substance for the biophysical behavior of the gel. The fluo-
Science + Business Media.) rescein concentration in the vitreous body can be estimated
by vitreous fluorophotometry. After intravenous (IV) injec-
tion of fluorescein, a certain amount (in healthy humans
only a very small amount) passes through the ocular barriers

168