Bilaminar Structure of the Human Optic Nerve Sheath.

PURPOSE/AIM: We aimed to characterize the connective tissue microanatomy, elastin abundance, and fiber orientation in the human optic nerve sheath, also known as the optic nerve dura mater, for correlation with its biomechanical properties. MATERIALS AND METHODS: Seven whole human orbits aged 4-93 years, and five isolated human optic nerve sheaths aged 26-75 years were formalin fixed, paraffin embedded, coronally sectioned, stained by Masson trichrome and van Gieson's elastin methods, and analyzed quantitatively for elastin fiber abundance and orientation. Elastin area fraction was defined as area stained for elastin divided by total area. RESULTS: While unilaminar in children, the adult ON sheath exhibited distinct inner and outer layers. Collagen was denser and more compact in the inner layer. Elastin area fraction was significantly greater at 6.0 ± 0.4% (standard error of mean) in the inner than outer layer at 3.6 ± 0.4% (P < 10-5). Elastin fibers had three predominant orientations: longitudinal, diagonal, and circumferential. Of circumferential fibers, 63 ± 4.7% were in the inner and 37 ± 4.7% in the outer layer (P < 10-4). Longitudinal and diagonal fibers were uniformly distributed in both layers. Elastin density and sheath thickness increased significantly with age (P < .01). CONCLUSIONS: The adult human optic nerve sheath is bilaminar, with each layer containing elastin fibers oriented in multiple directions consistent with isotropic properties. Differences in laminar elastin density and orientation may reflect greater tensile loading in the inner than in the outer layer.

Bilaminar Mechanics of the Human Optic Nerve Sheath.

PURPOSE/AIM: The adult human optic nerve (ON) sheath has recently been recognized to be bilaminar, consisting of inner layer (IL) and outer layer (OL). Since the ON and sheath exert tension on the globe in large angle adduction as these structures transmit reaction force of the medial rectus muscle to the globe, this study investigated the laminar biomechanics of the human ON sheath. MATERIALS AND METHODS: Biomechanical characterization was performed in ON sheath specimens from 12 pairs of fresh, post-mortem adult eyes. Some ON sheath specimens were tested completely, while others were separated into IL and OL. Uniaxial tensile loading under physiological temperature and humidity was used to characterize a linear approximation as Young's modulus, and hyperelastic non-linear behavior using the formulation of Ogden. Micro-indentation was performed by imposing small compressive deformations with small, hard spheres. Specimens of the same sheaths were paraffin embedded, sectioned at 10 micron thickness, and stained with van Gieson's stain for anatomical correlation. RESULTS: Mean (± standard error of the mean, SEM) tensile Young's modulus of the inner sheath at 19.8 ± 1.6 MPa significantly exceeded that for OL at 9.7 ± 1.2 MPa; the whole sheath showed intermediate modulus of 15.4 ± 1.1 MPa. Under compression, the inner sheath was stiffer (7.9 ± 0.5 vs 5.2 ± 0.5 kPa) and more viscous (150.8 ± 10.6 vs 75.6 ± 6 kPa s) than outer sheath. The inner sheath had denser elastin fibers than outer sheath, correlating with greater stiffness. CONCLUSIONS: We conclude that maximum tensile stiffness occurs in the elastin-rich ON sheath IL that inserts near the lamina cribrosa where tension in the sheath exerted during adduction tethering may be concentrated adjacent the ON head.

Compartmental Innervation of the Superior Oblique Muscle in Mammals.

PURPOSE: Intramuscular innervation of mammalian horizontal rectus extraocular muscles (EOMs) is compartmental. We sought evidence of similar compartmental innervation of the superior oblique (SO) muscle. METHODS: Three fresh bovine orbits and one human orbit were dissected to trace continuity of SO muscle and tendon fibers to the scleral insertions. Whole orbits were also obtained from four humans (two adults, a 17-month-old child, and a 33-week stillborn fetus), two rhesus monkeys, one rabbit, and one cow. Orbits were formalin fixed, embedded whole in paraffin, serially sectioned in the coronal plane at 10-µm thickness, and stained with Masson trichrome. Extraocular muscle fibers and branches of the trochlear nerve (CN4) were traced in serial sections and reconstructed in three dimensions. RESULTS: In the human, the lateral SO belly is in continuity with tendon fibers inserting more posteriorly on the sclera for infraducting mechanical advantage, while the medial belly is continuous with anteriorly inserting fibers having mechanical advantage for incycloduction. Fibers in the monkey superior SO insert more posteriorly on the sclera to favor infraduction, while the inferior portion inserts more anteriorly to favor incycloduction. In all species, CN4 bifurcates prior to penetrating the SO belly. Each branch innervates a nonoverlapping compartment of EOM fibers, consisting of medial and lateral compartments in humans and monkeys, and superior and inferior compartments in cows and rabbits. CONCLUSIONS: The SO muscle of humans and other mammals is compartmentally innervated in a manner that could permit separate CN4 branches to selectively influence vertical versus torsional action.

Nonclassical innervation patterns in mammalian extraocular muscles.

PURPOSE: The abducens (CN6) and oculomotor (CN3) nerves (nn) enter target extraocular muscles (EOMs) via their global surfaces; the trochlear (CN4) nerve enters the superior oblique (SO) muscle on its orbital surface. Motor nn are classically described as entering the EOMs in their middle thirds. We investigated EOM innervation that does not follow the classic pattern. METHODS: Intact, whole orbits of two humans and one each monkey, cow, and rabbit were paraffin embedded, serially sectioned in coronal plane, and prepared with Masson's trichrome and by choline acetyltransferase (ChAT) immunohistochemistry. Nerves innervating EOMs were traced from the orbital apex toward the scleral insertion, and some were reconstructed in three dimensions. RESULTS: Classical motor nn positive for ChAT entered rectus and SO EOMs and coursed anteriorly, without usually exhibiting recurrent branches. In every orbit, nonclassical (NC) nn entered each EOM well posterior to classical motor nn. These NC nn entered and arborized in the posterior EOMs, mainly within the orbital layer (OL), but often traveled into the global layer or entered an adjacent EOM. Other NC nn originated in the orbital apex and entered each EOM through its orbital surface, ultimately anastomosing with classical motor nn. Mixed sensory and motor nn interconnected EOM spindles. CONCLUSIONS: EOMs exhibit a previously undescribed pattern of NC innervation originating in the proximal orbit that partially joins branches of the classical motor nn. This NC innervation appears preferential for the OL, and may have mixed supplemental motor and/or proprioceptive functions, perhaps depending upon species. The origin of the NC innervation is currently unknown.

Characterization of ocular tissues using microindentation and hertzian viscoelastic models.

PURPOSE: The authors applied a novel microindentation technique to characterize biomechanical properties of small ocular and orbital tissue specimens using the hertzian viscoelastic formulation, which defines material viscoelasticity in terms of the contact pressure required to maintain deformation by a harder body. METHODS: They used a hard spherical indenter having 100 nm displacement and 100 µg force precision to impose small deformations on fresh bovine sclera, iris, crystalline lens, kidney fat, orbital pulley tissue, and orbital fatty tissue; normal human orbital fat, eyelid fat, and dermal fat; and orbital fat associated with thyroid eye disease. For each tissue, stress relaxation testing was performed using a range of ramp displacements. Results for single displacements were used to build quantitative hertzian models that were, in turn, compared with behavior for other displacements. Findings in orbital tissues were correlated with quantitative histology. RESULTS: Viscoelastic properties of small specimens of orbital and ocular tissues were reliably characterized over a wide range of rates and displacements by microindentation using the hertzian formulation. Bovine and human orbital fatty tissues exhibited highly similar elastic and viscous behaviors, but all other orbital tissues exhibited a wide range of biomechanical properties. Stiffness of fatty tissues tissue depended strongly on the connective tissue content. CONCLUSIONS: Relaxation testing by microindentation is a powerful method for characterization of time-dependent behaviors of a wide range of ocular and orbital tissues using small specimens, and provides data suitable to define finite element models of a wide range of tissue interactions.

Intramuscular innervation of primate extraocular muscles: unique compartmentalization in horizontal recti.

PURPOSE: It has been proposed that the lateral rectus (LR), like many skeletal and craniofacial muscles, comprises multiple neuromuscular compartments subserving different physiological functions. To explore the anatomic potential of compartmentalization in all four rectus extraocular muscles (EOMs), evidence was sought of possible regional selectivity in intramuscular innervation of all rectus EOMs. METHODS: Whole orbits of two humans and one macaque monkey were serially sectioned at 10 µm thickness and stained with Masson's trichrome. Three-dimensional reconstruction was performed of the intramuscular courses of motor nerves from the deep orbit to the anterior extents of their arborizations within all four rectus EOMs in each orbit. RESULTS: Findings concorded in monkey and human orbits. Externally to the global surface of the lateral (LR) and medial rectus (MR) EOMs, motor nerve trunks bifurcated into approximately equal-sized branches before entering the global layer and observing a segregation of subsequent arborization into superior zones that exhibited minimal overlap along the length of the LR and only modest overlap for MR. In contrast, intramuscular branches of the superior and the nasal portion of the inferior rectus were highly mixed. CONCLUSIONS: Consistent segregation of intramuscular motor nerve arborization suggests functionally distinct superior and inferior zones within the horizontal rectus EOMs in both humans and monkeys. Reduced or absent compartmentalization in vertical rectus EOMs supports a potential functional role for differential innervation in horizontal rectus zones that could mediate previously unrecognized vertical oculorotary actions.

Compartmentalized innervation of primate lateral rectus muscle.

PURPOSE: Skeletal and craniofacial muscles are frequently composed of multiple neuromuscular compartments that serve different physiological functions. Evidence of possible regional selectivity in LR intramuscular innervation was sought in a study of the anatomic potential of lateral rectus (LR) muscle compartmentalization. METHODS: Whole orbits of two humans and five macaque monkeys were serially sectioned at 10-microm thickness and stained with Masson trichrome. The abducens nerve (CN6) was traced anteriorly from the deep orbit as it branched to enter the LR and arborized among extraocular muscle (EOM) fibers. Three-dimensional reconstruction was performed in human and monkey orbits. RESULTS: Findings were in concordance in the monkey and human orbits. External to the LR global surface, CN6 bifurcated into approximately equal-sized trunks before entering the global layer. Subsequent arborization showed a systematic topography, entering a well-defined inferior zone 0.4 to 2.5 mm more posteriorly than branches entering the largely nonoverlapping superior zone. Zonal innervation remained segregated anteriorly and laterally within the LR. CONCLUSIONS: Consistent segregation of intramuscular CN6 arborization in humans and monkeys suggests functionally distinct superior and inferior zones for the LR. Since the LR is shaped as a broad vertical strap, segregated control of the two zones could activate them separately, potentially mediating previously unappreciated but substantial torsional and vertical oculorotary LR actions.

Effects of intracranial trochlear neurectomy on the structure of the primate superior oblique muscle.

PURPOSE. Although cyclovertical strabismus in humans is frequently attributed to superior oblique (SO) palsy, anatomic effects of SO denervation have not been studied. Magnetic resonance imaging (MRI) and orbital histology was used to study the effects of acute trochlear (CN4) denervation on the monkey SO. METHODS. Five juvenile macaque monkeys were perfused with formalin for 5 weeks: 15 months after unilateral or bilateral 10-mm intracranial trochlear neurectomy. Denervated and fellow orbits were imaged by MRI, embedded whole in paraffin, serially sectioned at 10-mum thickness, and stained with Masson trichrome. Whole muscle and individual fiber cross sections were quantified in SO muscles throughout the orbit and traced larger fibers in one specimen where they were present. RESULTS. MRI demonstrated marked reduction in midorbital cross section in denervated SO muscles, with anterior shift of SO mass preserving overall volume. Muscle fibers exhibited variable atrophy along their lengths. Denervated orbital layer (OL) fiber cross sections were slightly but significantly reduced from control at most anteroposterior locations, but this reduction was much more profound in global layer (GL) fibers. Intraorbital and intramuscular CN4 were uniformly fibrotic. In one animal, there were scattered clusters of markedly hypertrophic GL fibers that exhibited only sparse myomyous junctions only anteriorly. CONCLUSIONS. CN4 denervation produces predominantly SO GL atrophy with relative OL sparing. Overall midorbital SO atrophy was evident by MRI as early as 5 weeks after denervation, as denervated SO volume shifted anteriorly. Occasional GL fiber hypertrophy suggests that at least some SO fibers extend essentially the full muscle length after trochlear neurectomy.

Fascicular specialization in human and monkey rectus muscles: evidence for anatomic independence of global and orbital layers.

PURPOSE: Connective tissue pulleys inflect the extraocular muscles (EOMs) and receive insertions from some fibers. The authors investigated insertions and anatomic relationships of fiber fascicles within rectus EOMs to clarify the relationship to their pulleys. METHODS: Two human and two monkey orbits were removed intact, serially sectioned in the coronal plane, histologically stained, and digitally photographed. The authors traced representative fascicles in the human medial rectus (MR) and inferior rectus and monkey lateral rectus and superior rectus muscles. In the human MR, the authors computed average collagen fractions in the orbital layer (OL) and the global layer (GL). RESULTS: In human and monkey, OL fascicles remained distinct from each other and from the GL throughout. Most OL fascicles were inserted into the pulley through short tendons. Most GL fascicles bypassed the pulley without insertion. Collagen content in the human MR OL increased from 29% +/- 5% (SD) in midorbit to 65% +/- 9% in the anterior orbit but slightly decreased from 26% +/- 6% to 23% +/- 1% in the GL. Tracing of every fiber in a human MR OL fascicle demonstrated terminations on pulley tendons without myomyous junctions. CONCLUSIONS: Fibers in the primate rectus OL lack myomyous or GL junctions, but nearly all insert on the pulley through a broad distribution of short tendons and dense intercalated collagen. Fibers in the GL generally do not insert on pulley tissues and are associated with less collagen. These features support the distinct role of the OL in anteroposterior positioning of connective tissues proposed in the active pulley hypothesis and substantial mechanical independence of the OL and GL.

Horizontal rectus muscle anatomy in naturally and artificially strabismic monkeys.

PURPOSE: Structural abnormalities of extraocular muscles (EOMs) or their pulleys are associated with some forms of human strabismus. This experiment was conducted to investigate whether such abnormalities are associated with artificial or naturally occurring strabismus in monkeys. METHODS: Binocular alignment and grating visual acuities were determined in 10 monkeys representing various species using search coil recording and direct observations. Four animals were orthotropic, two had naturally occurring "A"-pattern esotropia, two had concomitant and one had "V"-pattern esotropia artificially induced by alternating or unilateral occlusion in infancy, and one had "A"-pattern exotropia artificially induced by prism wear. After euthanasia, 16 orbits were examined by high-resolution magnetic resonance imaging (MRI) in the quasi-coronal plane. Paths and sizes of horizontal rectus EOMs were analyzed quantitatively in a standardized coordinate system. Whole orbits were then serially sectioned en bloc in the quasi-coronal plane, stained for connective tissue, and compared with MRI. Nerve and EOM features were analyzed quantitatively. RESULTS: Quantitative analysis of MRI revealed no significant differences in horizontal rectus EOM sizes or paths among orthotropic or naturally or artificially strabismic monkeys. Histologic examination demonstrated no differences in EOM size, structure, or innervation among the three groups, and no differences in connective tissues in the pulley system. The accessory lateral rectus (ALR) EOM was present in all specimens, but was small, inconsistently located, and sparsely innervated. Characteristics of the ALR did not correlate with strabismus. CONCLUSIONS: Major structural abnormalities of horizontal rectus EOMs and associated pulleys are unrelated to natural or artificial horizontal strabismus in the monkeys studied. The ALR is unlikely to contribute to horizontal strabismus in primates. However, these findings do not exclude a possible role of pulley abnormalities in disorders such as cyclovertical strabismus.

Fatal toxic shock syndrome associated with Clostridium sordellii after medical abortion.

Endometritis and toxic shock syndrome associated with Clostridium sordellii have previously been reported after childbirth and, in one case, after medical abortion. We describe four deaths due to endometritis and toxic shock syndrome associated with C. sordellii that occurred within one week after medically induced abortions. Clinical findings included tachycardia, hypotension, edema, hemoconcentration, profound leukocytosis, and absence of fever. These cases indicate the need for physician awareness of this syndrome and for further study of its association with medical abortion.

Superior oblique muscle layers in monkeys and humans.

PURPOSE: Rectus and the inferior oblique extraocular muscles (EOMs) consist of orbital layers (OLs), inserting on connective tissues, and global layers (GLs), inserting on the sclera. This study was performed to clarify the anatomic relationships of the corresponding layers of the superior oblique (SO) muscle. METHODS: Two whole human and two monkey orbits were serially sectioned en bloc at 10-mum thickness in the coronal plane and stained for collagen with Masson's trichrome and for elastin with van Gieson's stain. The SO muscles of one human and one monkey were sectioned longitudinally. The structure of the SO muscle was examined by light microscopy, and muscle fibers in the OL and GL of selected sections were counted. RESULTS: The deep SO muscle consisted of a central GL contiguous with the tendon, surrounded coaxially by a peripheral OL inserting on the SO sheath posterior to the trochlea. The maximum number of SO fibers was 14,400 to 19,200 in the human and 7,000 to 7,400 in the monkey. In the monkey, approximately 60% of total fibers were in the GL, and 40% in the OL. The SO sheath was in mechanical continuity with the superior rectus pulley. CONCLUSIONS: The primate SO has a substantial OL configured to contribute to positioning the superior rectus pulley in the coronal plane. Whereas the direction of application of the SO's GL force is determined by the rigid trochlea, the SO's OL influences the direction of application of rectus EOM forces. This insight extends the concept of active control of pulley positions to include a contribution from the SO muscle.

Demonstration of systematic variation in human intraorbital optic nerve size by quantitative magnetic resonance imaging and histology.

PURPOSE: To use magnetic resonance imaging (MRI) to measure the diameter along the course of the intraorbital optic nerve in living subjects and cadaveric specimens, and to validate measurements histologically in the same specimens. METHODS: Measurements of the intraorbital optic nerve were made in 23 living human subjects and in three formalin-fixed orbits using high-resolution, surface coil MRI in the coronal plane. Fixed orbits were then serially sectioned in the same plane, and stained by Masson's trichrome for digital morphometry of nerve diameter and densitometry of connective tissue constituents. RESULTS: In cadaveric specimens, MRI and histologic measurements of optic nerve dimensions were in close quantitative agreement, showing significant decrease in the average optic nerve diameter along its retrobulbar course. This finding was confirmed by MRI in living subjects, with average optic nerve diameter declining from 3.99 +/- 0.04 mm (SEM) just posterior to the globe, to 3.50 +/- 0.04 mm at 10 mm further to the posterior (P < 0.0001). Color densitometry demonstrated a consistent cross-sectional area of nervous tissue, but decreasing amounts of collagen posteriorly. CONCLUSIONS: There is a significant decrease in normal optic nerve diameter along its length in normal subjects, reflecting reduction in connective tissue. High-resolution MRI is a valid and sensitive method of detecting subtle changes in retrobulbar optic nerve size and can be useful in the investigation of structural optic nerve lesions. Optic nerve diameters must be measured, however, at similar distances posterior to the globe to allow meaningful comparisons.

Evidence for a pulley of the inferior oblique muscle.

PURPOSE: This study was undertaken to investigate evidence for a connective tissue pulley constraining the path of the inferior oblique (IO) muscle. METHODS: From magnetic resonance images, the cross-sectional area, path, and orbital relationships of the human IO were determined in multiple gaze positions. Rectus pulleys were directly imaged with intravenous gadodiamide contrast. Images were compared with serial histologic sections of IO muscles of humans and monkeys. RESULTS: The IO path from origin to the lateral border of the inferior rectus (IR) muscle was straight. Lateral to the IR, the IO curved to follow the globe. At the point of IR crossing, the IO moved anteriorly from infraduction to supraduction by approximately 53% of the IR insertion's travel. Gaze-related change in IO cross section was demonstrable near the IR center. The gaze-related inflection in IO path corresponded to its encirclement by a pulley consisting of a dense ring of collagen, stiffened by elastin and smooth muscle, and united with the IR pulley. Orbital layer fibers of the IO inserted on its pulley, the lateral rectus (LR) pulley, and associated connective tissues. CONCLUSIONS: Like the rectus muscles, the human and monkey IO has a connective tissue pulley serving as its functional origin. The position of the IO pulley is influenced by its coupling to the actively moving IR pulley, whereas in turn the IO orbital layer inserts on and presumably shifts the IR and LR pulleys. These intercouplings facilitate implementation by rectus extraocular muscle suspensions of a commutative ocular motor plant.

Extraocular connective tissue architecture.

Extraocular muscle pulleys, now well known to be kinematically significant extraocular structures, have been noted in passing and described in fragments several times over the past two centuries. They were late to be fully appreciated because biomechanical modeling of the orbit was not available to derive their kinematic consequences, and because pulleys are distributed condensations of collagen, elastin and smooth muscle (SM) that are not sharply delineated. Might other mechanically significant distributed extraocular structures still be awaiting description?An imaging approach is useful for describing distributed structures, but does not seem suitable for assessing mechanical properties. However, an image that distinguished types and densities of constituent tissues could give strong hints about mechanical properties. Thus, we have developed methods for producing three dimensional (3D) images of extraocular tissues based on thin histochemically processed slices, which distinguish collagen, elastin, striated muscle and SM. Overall tissue distortions caused by embedding for sectioning, and individual-slice distortions caused by thin sectioning and subsequent histologic processing were corrected by ordered image warping with intrinsic fiducials. We describe an extraocular structure, partly included in Lockwood's ligament, which contains dense elastin and SM bands, and which might refine horizontal eye alignment as a function of vertical gaze, and torsion in down-gaze. This active structure might therefore be a factor in strabismus and a target of therapeutic intervention.

Quantitative analysis of the structure of the human extraocular muscle pulley system.

PURPOSE: Extraocular muscle (EOM) paths are constrained by connective tissue pulleys serving as functional origins. The quantitative structural features of pulleys and their intercouplings and orbital suspensions remain undetermined. This study was designed to quantify the composition of EOM pulleys and suspensory tissues. METHODS: Five human orbits, ages 33 weeks gestation to 93 years, were imaged intact by magnetic resonance (MRI), serially sectioned at 10 micro m thickness, and stained for collagen, elastin, and smooth muscle (SM). With MRI used as a reference, digital images of sections were geometrically corrected for shrinkage and processing deformations, and normalized to standard normal adult globe diameter. EOM pulleys, interconnections, suspensory tissues, and entheses were quantitatively analyzed for collagen, elastin, and SM thickness and density. RESULTS: Rectus and inferior oblique pulleys had uniform structural features in all specimens, comprising a dense EOM encirclement by collagen 1 to 2 mm thick. Elastin distribution varied, but was greatest in the orbital suspension of the medial rectus pulley and in a band from it to the inferior rectus pulley. This region corresponded to maximum SM density. Structural features of pulleys, intercouplings, and entheses were similar among specimens. The major mechanical couplings to the osseous orbit were near the medial and lateral rectus pulleys. CONCLUSIONS: Quantitative analysis of structure and composition of EOM pulleys and their suspensions is consistent with in vivo MRI observations showing discrete inflections in EOM paths that shift predictably with gaze. Focal SM distributions in the suspensions suggest distinct roles in stiffening as well as shifting rectus pulleys.