Equine Laminitis

Christopher C. Pollitt, BVSc, PhD

Laminitis, failure of the distal phalanx to maintain its attachment to the lamellae of the
inner hoof wall, causes unrelenting pain and a characteristic lameness. During a
developmental phase, pathology in organs anatomically remote from the foot generates
laminitis trigger factors that circulate to cause separation and disorganization of hoof
lamellar anatomy. Laminitis, linked to seasonal variations in grass fructan concentra-
tion can be induced experimentally using oligofructose, a closely related compound.
There is a strong correlation between the degree of lameness and the severity of
histopathology in lamellar samples from horses with laminitis. Matrix metalloproteinase
enzymes (MMPs), responsible for normal enzymatic remodeling of the epidermal
lamellae, appear to be accidentally recruited in the pathogenesis of the laminitis.
MMP-2 and MMP-9 have been isolated from normal lamellar tissues and in increased
quantities from lamellar tissues affected by laminitis. Although epidermal cells of other
species readily increase their production of MMP when exposed to cytokines this is not
the case with equine lamellar explants. An enzymatic theory of laminitis etiology based
on lamellar MMP activation challenges the alternative view that laminitis develops
because of vascular pathology affecting the circulation of the foot. Epidermal cell
necrosis, intravascular coagulation, edema and other evidence of ischemia cannot be
identified in tissue affected by early carbohydrate-induced laminitis. In fact, laminitis
does not occur if the foot is in a state of vasoconstriction during the developmental
phase suggesting that exogenous trigger factors cause laminitis when they reach the
lamellar tissues via dilated blood vessels. Indeed acute laminitis is prevented in a single
cooled limb while laminitis develops in the three remaining limbs maintained at room
temperature. Small explants of cultured, hoof lamellar tissue, an in vitro model for
laminitis, show that a substance(s) (a potential exogenous laminitis trigger factor) in the
supernatant of cultures of Streptococcus bovis activates equine hoof MMP-2 and
causes lamellar separation. This is taken as evidence for a bacterial pathogenesis of
laminitis since the population of S. bovis, the microorganism responsible for rapid
fermentation of carbohydrate in the equine hindgut, explodes exponentially during the
developmental phase. Chemical inhibitors block the activity of the laminitis MMPs in
vitro and have the potential to prevent field cases of laminitis. At acute laminitis onset
many of the ultrastructural plaques (hemidesmosomes, HDs) that attach lamellar basal
cells to the basement membrane of the connective tissue of the distal phalanx, are
absent or disrupted. This is accompanied by BM separation, cytoskeleton damage and
rounding of basal cell nuclei. The magnitude of HD loss in lamellar basal cells, affected
by laminitis directly correlates to the dose of carbohydrate used to induce it. Sound
lamellar architecture depends on the structural integrity of HDs, a fact illustrated when
a newborn foal, lacking in plectin (one of the HD intracytoplasmic plaque proteins),
developed laminitis as soon as it was ambulatory. The weight-bearing basement mem-
brane of the hoof lamellar dermal epidermal interface is unique to the anatomy of
digitigrade equids. However the lamellae are dependent on MMP enzymes for tissue
remodeling thus making horses susceptible in situations that accidentally precipitate
MMP activation, HD failure, basement membrane disadhesion and ultimately laminitis.
Clin Tech Equine Pract 3:34-44 © 2004 Elsevier Inc. All rights reserved.

KEYWORDS equine laminitis, matrix metalloproteinase (MMP), basement membrane, Strepto-

coccus bovis, hemidesmosome, fructan

School of Veterinary Science, The University of Queensland, Australia.

Address reprint requests to C.C. Pollitt, BVSc, PhD, The University of Queensland, Department of Companion Animal Science, School of Veterinary Science,
Brisbane, Queensland 4072, Australia. E-mail: [email protected]

34 1534-7516/04/$-see front matter © 2004 Elsevier Inc. All rights reserved.

doi:10.1053/j.ctep.2004.07.003
Equine laminitis 35

Figure 1 Horse with severe laminitis in both front feet showing typical laminitis gait. The hind feet are placed as far
forward as possible before the horse attempts painful shuffling steps in front.

Figure 2 Sagittal section of a foot with severe chronic laminitis and a large lamellar wedge. The attachment between the distal
phalanx (DP) and the dorsal hoof wall (DHW) has failed and hoof and bone are now widely separated. The dashed yellow line
shows the original position of the distal phalanx. The solid black line shows that the distal phalanx has rotated (in the
direction of the curved black arrow) off the normally straight axis of the proximal and middle phalanges. The material now
between the inner hoof wall and the bone is abnormal and consists of epidermal tissue proliferating to form a weak,
disorganized mass called the lamellar wedge (yellow line). The descent of the unattached distal phalanx into the hoof capsule
has distorted the growth of the proximal hoof wall tubules and has caused the sole to become convex instead of concave
(dropped sole). Two dark hemorrhagic zones (white arrows) show the sites of greatest pressure and trauma.
36 C.C. Pollitt

Figure 3 Lateromedial radiograph of a digit with severe chronic laminitis. The distal phalanx (DP) has dropped deeply
into the hoof capsule and is close to the ground surface which is shown by the horizontal radiopaque marker. The hoof
distal phalanx distance (HDPD) is 42% of L the length of the palmar cortex (L) of the distal phalanx. The HDPD in the
normal horse is approximately 25% of L. There is a linear radiolucency beneath the hoof wall (arrowheads) indicating
that the epidermal lamellae of the inner hoof wall have separated from the dermal lamellae.

L aminitis is the most serious disease of the equine hoof and

causes pathological changes in anatomy that lead to dev-
astating loss of function. The simplest definition of laminitis
from the foot result in the lamellar tissues of the feet being
exposed to factors which lead to separation and disorganiza-
tion of lamellar anatomy. The exact nature of the laminitis
is: failure of the attachment between the distal phalanx and trigger factors, apparently reaching the lamellar tissues via
the inner hoof wall. A horse has laminitis when the lamellar the circulation, has yet to be elucidated. Sometimes no de-
architecture of the inner hoof wall, which normally suspends velopmental phase can be recognized: the horse or pony is
the distal phalanx from the inner surface of the hoof capsule, discovered in the acute phase with no apparent ill-health or
fails. Without the distal phalanx properly attached to the inciting problem occurring beforehand. Obesity and related
inside of the hoof, the weight of the horse and the forces of endocrinopathic problems have recently been incriminated
locomotion drive the bone down into the hoof capsule, in the pathogenesis of this insidious form of laminitis.5,6
shearing and damaging arteries and veins, crushing the co- Grass founder can also appear without warning and this has
rium of the sole and coronet, causing unrelenting pain and a now been linked to seasonal variations in the concentration
characteristic lameness (Fig. 1). of the soluble sugar fructan by temperate pasture species.7,8
Fructan can suddenly reach very high concentrations in the
stems of grass and trigger a laminitis-inducing gastrointesti-
The Phases of Laminitis nal disturbance when consumed by horses and ponies. That
A developmental phase, during which lamellar separation is laminitis can be induced by such sugars has been verified
triggered, precedes the appearance of the foot pain (the acute experimentally using oligofructose, a closely related com-
phase) of laminitis. This may be as short as 8 to 12 hours in pound.9 The parenteral injection of potent long acting corti-
the case of laminitis caused by exposure to the water-soluble costeroid preparations for the treatment of skin disease may
toxins of black walnut (Juglans nigra) heartwood shavings1 or precipitate iatrogenic acute laminitis.10
30 to 40 hours in the case of excessive ingestion of high starch The developmental phase merges into the acute phase of
grain.2-4 During the developmental phase and before the clin- laminitis which lasts from the onset of clinical foot pain and
ical appearance of foot pain the horse or pony usually expe- lameness at the trot, to the time when there is clinical (usually
riences a problem with one or more of the following organ radiological) evidence of displacement of the distal phalanx
systems: gastrointestinal, respiratory, reproductive, renal, within the hoof capsule (Figs. 2 and 3). After the acute phase,
endocrine, musculoskeletal, integumentary and immune. if the horse does not die from the disease process inciting the
Multi-systemic aberrations in organs anatomically remote development of laminitis, it can make an apparent complete
Equine laminitis 37

Figure 4 Grade 1 histological laminitis (PAS stain). Micrograph showing hoof lamellar tissues stained to highlight the
basement membrane. The basement membrane (arrowed) is stained dark magenta. At the now tapered tips of the
secondary epidermal lamellae (SELs) the basement membrane has lifted away (stars) from the underlying basal cells.
Between the SEL bases the BM is in its normal position, close to the primary epidermal lamella (PEL). PAS stain. Bar ⫽
10 ␮m.

recovery or develop palmar/plantar displacement of the distal cated. The use of nerve blocks to eliminate pain will also
phalanx, the hallmark of chronic laminitis. The chronic phase encourage locomotion and precipitate more damage.
can last indefinitely with clinical signs ranging from persis-
tent, mild lameness, continued severe foot pain, further de-
generation of lamellar attachments, recumbency, hoof wall The Laminitis Histological
deformation and sloughing of the hooves.11 It is important to Grading System
realize that the process initiating the destruction of the lamel-
lar attachment apparatus begins to operate during the devel- As laminitis develops a sequence of histopathological
opmental phase before the first clinical sign of laminitis, foot changes occurs. Three grades of histological laminitis have
pain, is apparent. During the developmental phase the spe- been identified based on the degree of severity of the changes.
cific problems of the horse often have to be attended to ur- Making the lamellar basement clearly visible is important and
gently (eg, acute abdomen, grain overload acidosis, electro- requires staining lamellar tissues with either periodic acid
lyte imbalance, rhabdomyolysis, retained placenta) and Schiff (PAS), or periodic acid silver methanamine (PASM)
unfortunately the feet are often left out of therapeutic equa- stains, or with immunohistochemical methods using base-
tion until the first signs of foot pain (shifting weight from one ment membrane (BM) specific antibodies.
foot to the other) appear. By the time foot pain is apparent
lamellar pathology is underway. In other words foot pain is
the clinical sign that lamellar disintegration is occurring. To Grade 1 Histological Laminitis
wait and see if foot pain is the sequel to a metabolic crisis is to During the developmental phase lamellar basal and parabasal
miss the opportunity to prevent or at least ameliorate lamellar cells lose their normal shape, become elongated and appear
pathology. There is a good correlation between the severity of to slide over one another and, as a consequence, the second-
laminitis histopathology, as seen with the microscope, and ary epidermal lamellae become attenuated with tapering, in-
the degree of lameness [using the Obel grading system2] stead of club-shaped, tips.3,12 While this is going on, the BM
shown by the horse.3 When a horse first starts to show lami- of the SEL loses its attachment to the basal cells. This is first
nitic pain, the anatomy of the hoof wall lamellae is being noticeable at the tips of the SELs where small teat-shaped
destroyed. The higher the lameness grade, the more severe bubbles of loose BM form. To render this detectable by light
the microscopic damage. Any activity that places stress on an microscopy the tissues should be stained with periodic acid
already weakened lamellar attachment apparatus (such as Schiff (PAS), or periodic acid silver methanamine (PASM)
forced exercise) causes further damage and is contraindi- stains (Fig. 4).
38 C.C. Pollitt

Figure 5 Micrograph showing hoof lamellar tissues (PAS stain) with histological Grade 2 laminitis. The basement
membrane is stained dark magenta. At the tips of the now pointed secondary epidermal lamellae (SELs) the basement
membrane (BM) has continued to lift from the underlying basal cells to form empty, teat-shaped, caps (arrowheads).
The BM has disappeared from adjacent SEL bases and there is little connective tissue and few capillaries between. The
lamellar BM is no longer close to the primary epidermal lamella (PEL). Bar ⫽ 10 ␮m.

Grade 2 Histological Laminitis distance between hoof and distal phalanx becomes measur-
With disadhesion occurring between the lamellar BM and the able in millimeters (Figs. 6, 7, and 8). This is manifest clini-
SEL basal cells, the BM is drawn further away with each cycle cally as the “sinker.” Since the BM is the key structure bridg-
of foot loading by the horse. The lamellar basement mem- ing the epidermis of the hoof to the connective tissue of the
brane is now absent between the bases of adjacent SELs.13,14 distal phalanx, it follows that the wholesale loss and disorga-
The BM retracts from between the SELs and takes with it nization of the lamellar BM inexorably leads to the failure of
secondary dermal lamellar (SDL) connective tissue and SDL hoof anatomy so characteristic of the chronic stage of lami-
capillaries (Fig. 5). The loss of these capillaries may explain nitis.
why resistance to blood flow was increased 3.5 times (the
bounding digital pulse) in horses during early laminitis15 and The Pathophysiology of
why blood appears to bypass the lamellar capillary bed
through dilated arteriovenous anastomoses in horses with Laminitis
acute laminitis.14 Both of these phenomena oocur after, and The spectacular disintegration of the lamellar attachment ap-
are thus a consequence of, the triggering of MMP production. paratus, initiated during the development phase of laminitis,
The epidermal basal cells that have lost their BM attachment compromises a normally robust and trouble-free hoof, distal
do not appear to undergo necrosis, at least initially, and phalanx attachment apparatus in a surprisingly short period
clump together to form amorphous, BM-free masses, on ei- of time. Logic suggests that it is a normally tightly controlled
ther side of the primary lamellar axis. metabolic process that is thrown into disarray to cause the
lamellar-specific lesion of laminitis during its developmental
Grade 3 Histological Laminitis phase.9
In laminitis, the worse case scenario is a rapid and total BM The enzymatic remodeling of the epidermal lamellae, as-
separation from all the epidermal lamellae. Sheets of BM peel sumed to be mandatory if the continually proliferating stra-
away to form aggregations of loose isolated BM in the con- tum medium of the hoof wall16 is to move past the stationary
nective tissue adjoining the lamellae. The epidermal lamellar distal phalanx, could be accidentally recruited in the patho-
cells are left as isolated columns with no connection whatso- genesis of the laminitis disease process. Enzymes capable of
ever with the dermal connective tissue. The lamellar tips slide destroying key components of the lamellar attachment appa-
away from the BM connective tissue attachments, at first mi- ratus have been isolated from normal lamellar tissues17 and in
croscopically, but as the degree of separation increases the increased quantities from lamellar tissues affected by lamini-
Equine laminitis 39

Figure 6 Grade 3 histological laminitis (immunostain). The basement membrane of a lamellar tip is highlighted by type
IV collagen immunostaining. The tip of the primary epidermal lamella (PEL) has completely detached from its
basement membrane. The PEL basal cells are now an unattached, amorphous mass. Collapsed tubes of basement
membrane, now empty of epidermal cells, are still attached to connective tissue (arrowheads). The PEL has already
moved 0.03 mm from its dermal compartment and soon the distance will be measurable, in millimeters on a radio-
graph. The inset shows a normal lamellar tip, immunostained the same way. Type IV collagen immunostain. Bars ⫽ 10
␮m.

tis.18 The enzymes are metalloproteinase-2 and metallopro- circulating cytokines or some other trigger factor is yet to be
teinase-9 (MMP-2 and MMP-9) also found in a wide range of established. Evidence from in vitro studies, using equine la-
human and animal remodeling tissues such as bone, joints mellar explants, suggests that lamellar MMPs are not acti-
and endometrium as well as in metastasizing malignant tu- vated by exposure to cytokines.21
mors.19 The enzymatic theory of laminitis etiology based on lamel-
It is assumed that lamellar MMP activity is constantly re- lar MMP activation challenges the alternative view that lami-
sponding to the stresses and strains of normal equine life as nitis develops because of vascular pathology affecting the
well as to constant growth. When called for, sufficient MMP circulation of the foot. A current theory is that venoconstric-
is manufactured locally, to release epidermal cell-to-cell, and tion and high hydrostatic interstitial fluid pressure (compart-
cell-to-basement membrane attachments, as required, to ment syndrome) impede the flow of blood in the lamellar
maintain the correct shape and orientation of the hoof lamel- microcirculation to cause ischemic necrosis of epidermal la-
lae. From time to time injury to the basement membrane mellae.15 Epidermal cell necrosis, intravascular coagulation
would require its lysis and reconstruction. The controlled and edema were not identified in sections made from tissue
release of specific MMP inhibitors keeps this remodeling pro- in the early stages of laminitis.3 The vessels in the primary
cess in equilibrium and the hoof lamellae and the hoof itself dermal lamellae, even the smallest, were generally dilated
slowly migrate past the stationary basal cells firmly attached without evidence of microvascular thrombi.22 Further, no
to their underlying basement membrane that is in turn at- abnormalities in the systemic coagulation and fibrinolytic
tached to the connective tissue of the distal phalanx. cascades are found in horses with carbohydrate-induced
The epidermal cells of other species have been shown to acute laminitis.23 The gross anatomical appearance of freshly
readily increase their production of MMP when exposed to dissected laminitis tissue is one of dryness. Sometimes the
cytokines. Cultures of human oral mucosal keratinocytes re- lamellae peel apart. Tissues affected by a compartment syn-
spond to the addition of tumor necrosis factor (TNF), inter- drome exude fluid.
leukin-1 (IL-1) and transforming growth factor-1 (TGF-1) by How do the trigger factors of laminitis reach the lamellae?
increasing production of MMP-9.20 Lamellar tissues affected There is now strong evidence from three independent exper-
by laminitis also increase transcription of MMP17 and pro- imental sources24-26 that the foot circulation during the de-
duce MMPs in their active forms18 but whether in response to velopmental phase of laminitis is dilated. Laminitis does not
40 C.C. Pollitt

Figure 7 Grade 3 histological laminitis (immunostain). Only remnants (arrowheads) of the basement membrane (BM)
remain between the now disorganized secondary epidermal lamellae. Most of the lamellar epidermal cells have
coalesced into an amorphous mass no longer effectively attached to any connective tissue. The remainder of the lamellar
BM lies free, in strands (arrowed), among the connective tissue of the primary dermal lamella (PDL); primary epidermal
lamella (PEL). Type IV collagen immunostain. Bar ⫽ 10 ␮m.

occur if the foot is in a state of vasoconstriction during the light of recent research is that the lamellar disintegration of
developmental phase suggesting that the trigger factors will laminitis is mediated by the uncontrolled release of excess
only cause laminitis if they reach the lamellar tissues via MMP.17
dilated blood vessels at a high enough concentration and over We have successfully developed an in vitro model18,21 for
a long enough time period. It follows that therapy aimed at equine laminitis using small explants of tissue taken from the
keeping the feet of horses in danger of developing laminitis as inner hoof wall of normal, freshly killed, abattoir horses.
cool as possible (and therefore vasoconstricted) is logical. Each explant consists of stratum medium, the lamellar layer
Indeed acute laminitis can be prevented in a single cooled and the sublamellar connective tissue. After incubation for 48
limb while laminitis develops in the three remaining limbs hours in tissue culture medium, plus the laminitis trigger
maintained at room temperature.27 Horses, unlike humans, factor under investigation, each explant is subjected to ten-
do not regard extremely cold feet as uncomfortable and can sion. The force required to separate epidermal from dermal
tolerate having their feet in iced water for 48 hours with no lamellae is recorded. When dermal– epidermal lamellar sep-
discomfort or ill effect.28 Scintigraphic studies comparing the aration occurs readily (as occurs in field cases of laminitis) we
circulation of iced feet versus normal showed profound va- consider the tissue to have developed in vitro laminitis. La-
soconstriction in the cold feet (80.5% of precooled values).29 mellar explants can be cultured for up to 7 days in normal
What are the laminitis trigger factors? Since the carbohy- medium and no lamellar separation occurs. It is virtually
drate overload model of laminitis is characterized by endo- impossible to separate normal lamellar explants. One event
toxin production it would seem a safe presumption that mac- that readily causes separation of lamellar explants is MMP
rophages in the peritoneal cavity and elsewhere in the body activation. The addition to the culture medium of the organo-
would be subject to endotoxin stimulation just as they are mercurial compound aminophenylmercuric acetate (APMA),
during other acute gastrointestinal diseases.30 Mononuclear a well known nonphysiological MMP activator, readily in-
phagocytes express tumor necrosis factor along with other duces explant lamellar separation. Treatment of lamellar ex-
cytokines such as interleukin within minutes of exposure to plants with APMA is the in vitro laminitis control against
endotoxin. The cytokine cascade originating from an acute which naturally occurring laminitis induction factors can be
abdomen is responsible for most of the pathological effects of measured. The presence or absence of MMP activation in
endotoxemia. However, laminitis has never been triggered by explant supernatants is detected zymographically using gel-
the experimental administration of endotoxin into the blood- atin polyacrylamide electrophoresis and all explant tissues
stream31 or the peritoneal cavity and the actual trigger factors are fixed and examined histologically. Histological sections
of laminitis remain unidentified. What appears certain in the show a clear zone of complete separation between the base-
Equine laminitis 41

Figure 8 Diagrams showing normal lamellar histology and three grades of laminitis histopathology in order of increasing
severity.

ment membrane and the basal cells of the epidermal lamellae. nitis.21 If it crosses the mucosal barrier of the hindgut and
This is a characteristic of in vitro laminitis and resembles the enters the circulation it may be a “cause” of laminitis (at least
basement membrane lesion of natural in vivo laminitis. in the carbohydrate overload model). In other words it may
We have used the in vitro laminitis explant model to in- be an exogenous laminitis trigger factor.
vestigate most of the proposed causes of equine laminitis. The activity of tissue MMPs has long been shown to cor-
The equine lamellae have tested resistant to virtually all relate strongly with the degree of malignancy and invasive-
known cytokines, tissue factors and prostaglandins. Gram- ness of lethal human tumors such as malignant melanoma,
negative bacterial endotoxin, extract of black walnut (Juglans breast and colon cancer. We have recently cloned the gene
nigra) and even anaerobic culture conditions fail to induce responsible for MMP-2 expression in lamellar hoof.17 Horses
lamellar separation or significant MMP activation. There is with acute laminitis show increased expression of the MMP-2
one notable exception however. A factor present in the su- gene, 48 hours after alimentary carbohydrate overload (Fig.
pernatant of cultures of Streptococcus bovis isolated from the 9). For mean MMP-2 gene expression to have doubled by the
equine cecum activates equine hoof MMP-2 and causes la- time lameness is manifest implies that the factors signaling
mellar separation.21 During grain overload S. bovis is the prin- the increased expression have been present for some time.
cipal microorganism responsible for the rapid fermentation This places perturbation of MMP equilibrium early in the
of carbohydrate to lactic acid in the equine hindgut. In the cascade of events leading to the foot pain of acute, clinical
presence of virtually unlimited substrate its population ex- laminitis. Indeed biopsies of lamellar tissue taken at 24, 36,
plodes exponentially. We are currently investigating the role and 48 hours (Wattle and Pollitt, unpublished data) all
of the S. bovis MMP activator in natural cases of equine lami- showed some of the histopathology described in the pub-
42 C.C. Pollitt

Figure 9 Graph (left) showing the significantly different (P ⬍ 0.01) mean values of MMP-2 expression between 4 normal
hooves and 18 laminitis-affected hooves. Polyacrylamide gel zymography (gel contains 0.1% gelatin) of lamellar
explants from a horse with laminitis (right). Lane 1: normal hoof explant supernatant. Lanes 2 and 3: laminitis fore hoof
explant supernatants. Lanes 4 and 5: laminitis hind hoof explant supernatants. Molecular weights are derived from
standards (not shown). There is a significant increase is the amount of active MMP 9 (82 kDa, black arrowhead) and
MMP2 (62 kDa, white arrowhead).

lished laminitis grading systems.3,32 At 24 hours lamellae had underlying basement membrane. In lamellar SEL samples
intact basement membranes but SELs were attenuated with taken at the onset of acute laminitis many HDs are absent or
round basal cell nuclei. At 36 hours, SEL attenuation had disrupted (Fig. 10). Loss and disruption of HDs is accompa-
progressed and SEL basal cells with rounded nuclei were nied by BM separation, cytoskeleton damage and rounding of
disorganized; SEL tips were pointed instead of rounded. the basal cell nucleus. Indeed the magnitude of HD loss in
Only at 48 hours was the BM not attached to SEL basal cells SEL basal cells affected by laminitis directly correlates to the
suggesting that the disadhesion process commenced some- dose of carbohydrate used to induce it;12 data that support a
where between 36 and 48 hours. However the molecular and bacterial pathogenesis of laminitis.21 The hypothesis is that
biochemical events contributing to BM disattachment, as ev- increasing amounts of carbohydrate substrate support
idenced by nuclear rounding and SEL attenuation, were in greater numbers of microbes that generate higher concentra-
place by 24 hours. The basement membrane lesion of lami- tions of laminitis trigger factors.
nitis is insidious in nature and well under way by the time Hoof lamellae cultured in vitro, separate under tension
clinicians are aware of laminitis foot pain. Any preventive33 or and the intracytoplasmic components of their HDs denucle-
treatment strategies must be in place before overt foot pain ate and fade if not provided with sufficient glucose;35 a mech-
develops if horses are to survive the development phase of anism that may be operating in vivo when toxemia and the
laminitis without significant lamellar damage. various endocrinopathies associated with laminitis limit the
There is a wide range of chemical agents capable of inhib- supply of lamellar glucose. Activation of constituent lamellar
iting MMP activity both in vitro and in vivo.34 We have MMPs also causes lamellar separation under tension but
shown that one of these (Batimastat or BB-94, British Biotech, without affecting HD ultrastructure. Activated MMPs appear
Oxford) blocks the activity of the laminitis MMPs in vitro and to cleave laminin5 anchoring filaments and set the BM adrift;
has the potential to be a useful tool in the prevention and also a process now shown to occur in vivo.32
management of acute laminitis.18 Trials to test whether MMP The dependence of sound lamellar architecture on the
inhibitors can prevent or ameliorate field cases of laminitis structural integrity of HDs is well illustrated when foals are
are currently underway in the Australian Equine Laminitis born lacking just one of their HD proteins. A 45-day-old
Research Unit at The University of Queensland. Quarterhorse foal showed the clinical signs and gross pathol-
ogy of chronic laminitis since birth.36 Characteristic laminitis
histopathology was present only in the front feet. In all feet
The Ultrastructure of Laminitis TEM showed the intracytoplasmic plaques of lamellar
Laminitis studied by transmission electron microscopy hemidesmosomes were small, misshapen and not associated
(TEM) and immunofluorescence microscopy (IFM) has pro- with the cytoskeleton. In all feet IFM showed the hemides-
vided new insight into the mechanism of the disease. The mosomal, intracytoplasmic plaque protein, plectin, was ab-
hemidesmosome (HD) is the attachment plaque responsible sent. The foal was a rare case of congenital epidermolysis
for maintaining contact between the SEL basal cell and its bullosa simplex, an inherited failure to express plectin. Lack-
Equine laminitis 43

Figure 10 Transmission electron micrographs (TEMs) of lamellar SELs at the onset of acute laminitis. In (A) many
hemidesmosomes (arrow) are absent or faded. Anchoring filaments (arrowhead) are present where hemidesmosomes
are still relatively normal. Loss and disruption of hemidesmosomes is accompanied by the commencement of BM
separation and damage of the cytoskeleton (*). Bar ⫽ 500 nm. In (B) the BM has separated from the attenuated, pointed
SEL tip and formed a typical, empty BM enclosed bubble. There are few recognizable hemidesmosomes and only
fragments of cytoskeleton (*). Bar ⫽ 200 nm. D, dermis; E, epidermal basal cell.

ing plectin, the cytoskeleton and hemidesmosomes of the Acknowledgments

hoof lamellae were unstable, resulting in laminitis when the This research was funded by grants from the RIRDC (Austra-
front feet first bore weight. The loss of even a single protein lia) entitled “Investigations into the cause and prevention of
causes structural failure and laminitis. The presence of histo- equine laminitis” and “High pasture fructan concentration
logical lesions only in the front feet illustrated the influence of causes equine laminitis.” In addition the author gratefully
weight distribution on laminitis severity and supports the acknowledges the generous financial assistance of the Animal
validity of preventive strategies that unload the feet. Health Foundation (St. Louis, MO) who have committed
Ideas on laminitis pathophysiology abound37 and this re- on-going funds to assist the work of the Australian Laminitis
view has focused on the MMP, enzymatic theory of laminitis Research Unit at The University of Queensland.
pathogenesis, a hypothesis that depends on the generation of
circulating toxins, or proinflammatory mediators (laminitis
trigger factors) in the gastrointestinal (carbohydrate over- References
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