Ultrastructural morphology is distinct among primary progenitor cell isolates from normal, inflamed, and cryopreserved equine hoof tissue and CD105+K14+ progenitor cells.

The equine hoof dermal-epidermal interface requires progenitor cells with distinct characteristics. This study was designed to provide accurate ultrastructural depictions of progenitor cells isolated from inflamed tissue and normal tissue before and after cryopreservation and following selection of cells expressing both keratin (K) 14 (ectodermal) and cluster of differentiation (CD) 105 (mesodermal). Passage 3 cell ultrastructure was assessed following 2D culture and after 3D culture on decellularized hoof tissue scaffolds. Outcome measures included light, transmission electron, and scanning electron microscopy, immunocytochemistry, and CD105+K14+ cell trilineage plasticity. Cells from normal tissue had typical progenitor cell characteristics. Those from inflamed tissue had organelles and morphology consistent with catabolic activities including lysosomes, irregular rough endoplasmic reticulum, and fewer vacuoles and early endosomes than those from normal tissue. Cryopreserved tissue cells appeared apoptotic with an irregular cell membrane covered by cytoplasmic protrusions closely associated with endocytic and exocytic vesicles, chromatin aggregated on the nuclear envelop, abundant, poorly organized rough endoplasmic reticulum, and plentiful lysosomes. Cells that were CD105+K14+ were distinguishable from heterogenous cells by infrequent microvilli on the cell surface, sparse endosomes and vesicles, and desmosomes between cells. Cells expressed ectodermal (K15) and mesodermal (CD105) proteins in 2D and 3D cultures. Inflamed and cryopreserved tissue isolates attached poorly to tissue scaffold while normal tissue cells attached well, but only CD105+K14+ cells produced extracellular matrix after 4 d. The CD105+K14+ cells exhibited osteoblastic, adipocytic, and neurocytic differentiation. Ultrastructural information provided by this study contributes to understanding of equine hoof progenitor cells to predict their potential contributions to tissue maintenance, healing, and damage as well post-implantation behavior.

Microtomographic Parameters and Nanoindentation of the Hoof of Girolando Cattle.

The aim of this study was to describe the microstructure of the pigmented and depigmented hoof capsule of Girolando cattle by bi- and tridimensional microtomography and nanoindentation, analysing the possible relation between these findings and the susceptibility of such animals to podal diseases. To carry out the microtomography and the nanoindentation, duplicated samples were collected from the dorsal wall, abaxial wall and pre-bulbar sole of the hoof capsule. Material collection was performed in 40 medial digits of thoracic limbs and 40 lateral digits of pelvic limbs. The bidimensional microtomography showed that the dorsal wall of the thoracic and pelvic limbs presented higher density, followed by the abaxial wall, and finally by the sole, with the lowest density. Moreover, the hoof capsule of cows of Girolando breed is a compact, non-porous material, and constituted by extratubular and intratubular keratin. By the tridimensional microtomography, it was possible to measure the angles of the corneal tubules in relation to the periople and the claws in the different regions of the hoof capsule, which were 90° for the dorsal wall, 55° for the abaxial wall and 70° for the sole. The tridimensional microtomography also showed corneal tubules of different diameters: 17, 51, 85, 119 and 153 µm. The nanoindentation test, when performed in different regions of the hoof capsule, did not reveal significant difference of Vickers hardness in the evaluated areas. However, we verified a larger elastic module of these regions on the transversal cut of the corneal tubules compared to the longitudinal cut.

Microstructure and Hardness of Buffalo's Hoofs.

The aim of this study was to describe the microstructure of hoof capsules of the buffalo. In addition, the study emphasized the morphometric aspects of the horn tubules, the Vickers nanohardness of the dorsal and abaxial walls and sole of the digits of the thoracic and pelvic limbs of the buffalo. The abaxial wall in the thoracic and pelvic digits showed larger diameter of the horn tubules when compared to all dorsal wall and sole. In addition, the abaxial wall of the thoracic digits showed larger diameter of the horn tubules when compared with the pelvic digits. According to the three-dimensional microtomography, the dorsal wall was higher in density compared with the abaxial wall. The latter exhibited an intermediate density, while the sole showed the lowest density. The Vickers nanohardness test showed that there was no difference in hardness and resistance between the experienced regions. However, the elastic modulus was greater on the transversal section of the hoof capsule. In conclusion, the results of the current study show that modern technologies such as microtomography and subsequent imaging can be used to investigate details of the basic morphology in different regions of the buffalo's hoof.

Comparative study of the claws of Pediculus humanus capitis between archaeological and modern specimens.

Metric data of the claws of archaeological specimens of Pediculus humanus capitis (dating between 1500 B.C. and A.D. 1500) and modern lice specimens coming from school children were analyzed and compared. Both sets of samples come from Arica in northern Chile. The overall sample is comprised of 14 archaeological specimens (6 females and 8 males) of Pediculus humanus capitis and 22 modern specimens (13 females and 9 males). All specimens were studied with scanning electron microscopy (SEM), uncoated, using variable pressure mode. The objective of this study was to metrically analyze the first couple of clutches of ancient and modern adult lice specimens (width and length of the tibio-tarsal claw and tarsus length) to test if morphological changes have taken place throughout time in these anatomical elements. We found that archaeological male and female specimens presented significant differences in the tibio-tarsal width (right and left). When comparing data between archaeological and modern male specimens, statistically significant differences were found in almost all the parameters studied, except for the right tarsal length. On the other hand, archaeological and modern female specimens showed no statistically significant change in the variables studied. In brief, our data suggest that modern male specimens have undergone a process of claw reduction, but females have maintained the same dimensions.

Filamentous Structure of Hard ß-Keratins in the Epidermal Appendages of Birds and Reptiles.

The structures of avian and reptilian epidermal appendages, such as feathers, claws and scales, have been modelled using X-ray diffraction and electron microscopy data, combined with sequence analyses. In most cases, a family of closely related molecules makes up the bulk of the appendage, and each of these molecules contains a central ß-rich 34-residue segment, which has been identified as the principal component of the framework of the 3.4 nm diameter filaments. The N- and C-terminal segments form the matrix component of the filament/matrix complex. The 34-residue ß-rich central domains occur in pairs, related by either a parallel dyad or a perpendicular dyad axis, and form a ß-sandwich stabilized by apolar interactions. They are also twisted in a right-handed manner. In feather, the filaments are packed into small sheets and it is possible to determine their likely orientation within the sheets from the low-angle X-ray diffraction data. The physical properties of the various epidermal appendages can be related to the amino acid sequence and composition of defined molecular segments characteristic of the chains concerned.

Ability of hydroxypropyl chitosan nail lacquer to protect against dermatophyte nail infection.

The development of a topical agent that would strengthen the nail, improve the natural barrier, and provide better drug penetration to the nail bed is needed. In this study, we examined the effects of a hydroxypropyl chitosan (HPCH)-based nail solution using a bovine hoof model. Following application of the nail solution, changes in the hardness of the hoof samples were measured using the Vickers method. Tensile and flexural strengths were tested by stretching or punching the samples, respectively. The ultrastructure was examined using scanning electron microscopy (SEM), and samples stained with periodic acid-Schiff (PAS) stain were used to determine the fungal penetration depth. The comparators included 40% urea and 70% isopropyl alcohol solutions. The HPCH nail solution increased hoof sample hardness in comparison to the untreated control sample (mean, 22.3 versus 19.4 Vickers pyramid number [HV]). Similarly, the HPCH solution increased the tensile strength (mean, 33.07 versus 28.42 MPa) and flexural strength (mean, 183.79 versus 181.20 MPa) compared to the untreated control. In contrast, the comparators had adverse effects on hardness and strength. SEM showed that the HPCH solution reduced the area of sample crumbling following abrasion compared to the untreated control (7,418 versus 17,843 pixels), and the PAS-stained images showed that the HPCH solution reduced penetration of the dermatophyte hyphae (e.g., penetration by Trichophyton mentagrophytes was <25 µm at day 9 versus 275 µm in the untreated control). Unlike chemicals normally used in cosmetic treatments, repeated application of the HPCH nail solution may help prevent the establishment of new or recurring fungal nail infection.

Comparative anatomy of mouse and human nail units.

Recent studies of mice with hair defects have resulted in major contributions to the understanding of hair disorders. To use mouse models as a tool to study nail diseases, a basic understanding of the similarities and differences between the human and mouse nail unit is required. In this study we compare the human and mouse nail unit at the macroscopic and microscopic level and use immunohistochemistry to determine the keratin expression patterns in the mouse nail unit. Both species have a proximal nail fold, cuticle, nail matrix, nail bed, nail plate, and hyponychium. Distinguishing features are the shape of the nail and the presence of an extended hyponychium in the mouse. Expression patterns of most keratins are similar. These findings indicate that the mouse nail unit shares major characteristics with the human nail unit and overall represents a very similar structure, useful for the investigation of nail diseases and nail biology.

Microanatomy of passerine hard-cornified tissues: beak and claw structure of the black-capped chickadee (Poecile atricapillus).

The microanatomy of healthy beaks and claws in passerine birds has not been well described in the literature, despite the importance of these structures in avian life. Histological processing of hard-cornified tissues is notoriously challenging and only a few reports on effective techniques have been published. An emerging epizootic of beak deformities among wild birds in Alaska and the Pacific Northwest region of North America recently highlighted the need for additional baseline information about avian hard-cornified structures. In this study, we examine the beak and claw of the Black-capped Chickadee (Poecile atricapillus), a common North American passerine that is affected by what has been described as "avian keratin disorder." We use light and scanning electron microscopy and high-magnification radiography to document the healthy microanatomy of these tissues and identify features of functional importance. We also describe detailed methods for histological processing of avian hard-cornified structures and discuss the utility of special stains. Results from this study will assist in future research on the functional anatomy and pathology of hard-cornified structures and will provide a necessary reference for ongoing investigations of avian keratin disorder in Black-capped Chickadees and other wild passerine species.

Hydration and N-acetyl-l-cysteine alter the microstructure of human nail and bovine hoof: implications for drug delivery.

This work aimed to (a) characterize the microstructure and porosity of human nail and bovine hoof by mercury intrusion porosimetry and SEM image analysis, (b) study the effects of hydration and of N-acetyl-l-cysteine treatment on the microstructure of both membranes, and (c) determine whether the microstructural modifications were associated with changes in drug penetration measured by standard diffusion studies. Bovine hoof surface is more porous than nail surface although there were no differences between the mean surface pore sizes. Hydration and N-acetyl-l-cysteine increased the roughness and apparent surface porosity, and the porosity determined by mercury intrusion porosimetry of both membranes. Pore-Cor™ was used to generate tridimensional structures having percolation characteristics comparable to nail and hooves. The modeled structures were horizontally banded having an inner less-porous area which disappeared upon treatment. Treatment increased the predicted permeability of the simulated structures. Triamcinolone permeation increased significantly for hooves treated N-acetyl-l-cysteine, i.e., the membranes for which microstructural and permeability changes were the largest. Thus, microstructural changes determined via mercury intrusion porosimetry and subsequently modeled by Pore-Cor™ were related to drug diffusion. Further refinement of the technique will allow fast screening of penetration enhancers to be used in ungual drug delivery.

Ultrastructural localization of hair keratin homologs in the claw of the lizard Anolis carolinensis.

The claw of lizards is largely composed of beta-keratins, also referred to as keratin-associated beta-proteins. Recently, we have reported that the genome of the lizard Anolis carolinensis contains alpha keratin genes homologous to hair keratins typical of hairs and claws of mammals. Molecular and immunohistochemical studies demonstrated that two hair keratin homologs named hard acid keratin 1 (HA1) and hard basic keratin 1 (HB1) are expressed in keratinocytes forming the claws of A. carolinensis. Here, we extended the immunocytochemical localization of the novel reptilian keratins to the ultrastructural level. After sectioning, claws were subjected to immunogold labeling using antibodies against HA1, HB1, and, for comparison, beta-keratins. Electron microscopy showed that the randomly organized network of tonofilaments in basal and suprabasal keratinocytes becomes organized in long and parallel bundles of keratin in precorneous layers, resembling cortical cells of hairs. Entering the cornified part of the claw, the elongated corneous cells fuse and accumulate corneous material. HA1 and HB1 are absent in the basal layer and lower spinosus layers of the claw and are expressed in the upper and precorneous layers, including the elongating corneocytes. The labeling for alpha-keratin was loosely associated with filament structures forming the fibrous framework of the claws. The ultrastructural distribution pattern of hard alpha-keratins resembled that of beta-keratins, which is compatible with the hypothesis of an interaction during claw morphogenesis. The data on the ultrastructural localization of hair keratin homologs facilitate a comparison of lizard claws and mammalian hard epidermal appendages containing hair keratins.

Characterisation of white line degeneration in sheep and evidence for genetic influences on its occurrence.

Shelly hoof in sheep occurs when the hoof wall becomes detached from the laminar corium; it often then becomes impacted with debris leading to infection, pain and lameness. The problem of shelly hoof is under-reported and is often confused with classical footrot. A study was conducted using data on 9,169 Blackface and Texel sheep from 22 farms in the UK. The aim of the study was to estimate the prevalence of shelly hoof in these breeds, to quantify the genetic basis to shelly hoof, and to describe a pilot trail to characterise the physical properties of horn using Transmission Electron Microscopy (TEM). The results showed that shelly hoof has a high prevalence (47% for Blackface and 24% for Texel ewes) and is under moderate genetic control (h(2) = 0.3). The TEM images showed fragmented dorsal wall horn, microscopic crevices leading deeper into the hoof accompanied by bacteria and other micro-organisms. The invasion of bacteria was facilitated by separation of the cells in a characteristic 'un-zipping' phenomenon, poor attachment of the keratin to the cell membranes, poor cell membrane quality with a 'lace-chain' effect and incomplete keratinisation of the cells.

Carbohydrate alimentary overload laminitis.

In acute laminitis, the suspensory apparatus of the distal phalanx fails at the lamellar dermal/epidermal interface. A grading system for the histopathology of laminitis is based on the consistent pattern of histologic changes to the secondary epidermal lamellae, basal cells, and basement membrane that occur as carbohydrate-induced laminitis develops. The actual trigger factors of carbohydrate-induced laminitis remain unidentified.

Scanning electron microscopy and fungal culture of hoof horn from horses suffering from onychomycosis.

Horn samples were taken from the hooves of eight horses with clinical signs of equine onychomycosis in at least one hoof capsule. None of the horses had a documented mycological history. The predominant alterations of the horn capsules were sand cracks, white line disease, brittleness (especially around the nail holes), parakeratosis and bruising. The horn samples were stored in sterile tubes for transportation and transferred onto Sabouraud Dextrose Agar and dermatophyte test agar for mycological examination within 6 h. Fungal cultures were incubated for 30 days at room temperature. Fungal identification was based on colonial morphology and microscopic examination of conidia. Horn samples were also stored at -80°C until used for scanning electron microscopy (SEM). The fungal culture revealed that the hoof horn from all eight horses was infected with keratinophilic fungi. The keratinopathogenic fungi Trichophyton spp and Scopulariopsis brevicaulis were also detected in six horses. SEM revealed severe alterations of the horn structure in horn samples infected with keratinopathogenic fungi compared to horn samples from a sound hoof. The most evident changes were deterioration of the tubular structure of the horn wall, disruption of the horny layers, superficial lysis of cornified cells and the presence of fungal elements. Samples without dermatophyte or Scopulariopsis infection, in contrast, were similar to healthy hoof horn.

Equine laminitis: ultrastructural lesions detected in ponies following hyperinsulinaemia.

REASONS FOR PERFORMING STUDY: Anatomical changes in the hoof lamellar tissue induced by prolonged hyperinsulinaemia have not been described previously. Analysis of the induced lesions may promote understanding of hyperinsulinaemic laminitis pathogenesis and produce clinical benefit. OBJECTIVES: To use light and transmission electron microscopy (TEM) to document hoof lamellar lesions in ponies clinically lame after prolonged hyperinsulinaemia. METHODS: Nine clinically normal, mature ponies were allocated randomly to either a treatment group (n = 5) or control group (n = 4). The treatment group received insulin via a modified, prolonged euglycaemic hyperinsulinaemic clamp technique (EHCT) and were subjected to euthanasia when clinical signs of Obel grade II laminitis occurred. The control group was sham treated with an equivalent volume of 0.9% saline and killed at 72 h. Lamellar tissues of the right front feet were harvested and processed for TEM. RESULTS: Lamellae from insulin treated ponies were attenuated and elongated with many epidermal basal cells (EBC) in mitosis. Unlike carbohydrate induced laminitis in horses there was no global separation at the lamellar dermal/epidermal interface among ponies. Sporadic EBC basement membrane (BM) separation was associated with the proximity of infiltrating leucocytes. In 2 ponies, the lamellar BM was thickened. The number of hemidesmosomes/microm of BM was decreased in all insulin treated ponies. CONCLUSIONS: Prolonged hyperinsulinaemia causes unique lamellar lesions normally characteristic of acute and chronic laminitis. Lamellar proliferation may be an insulin effect through its mitogenic pathway. Aberrant lamellar mitosis may lengthen and weaken the lamellar, distal phalanx attachment apparatus and contribute to the clinical signs that developed. POTENTIAL RELEVANCE: The study shows that insulin alone, in higher than normal circulating concentrations, induces profound, changes in lamellar anatomy. Medical control of insulin resistance and hyperinsulinaemia may ameliorate lesions and produce clinical benefit.

Biomechanics of dromaeosaurid dinosaur claws: application of X-ray microtomography, nanoindentation, and finite element analysis.

Dromaeosaurid theropod dinosaurs, such as Velociraptor, possess strongly recurved, hypertrophied and hyperextensible ungual claws on the pes (digit II) and manus. The morphology of these unguals has been linked to the capture and despatching of prey. However, the mechanical properties or, more importantly, the mechanical potential of these structures have not been explored. Generation of a 3D finite element (FE) stress/strain contour map of a Velociraptor manual ungual has allowed us to evaluate quantitatively the mechanical behavior of a dromaeosaurid claw for the first time. An X-ray microtomography scan allowed construction of an accurate 3D FE mesh. Analogue material from an extant avian theropod, the pedal digit and claw of an eagle owl (Bubo bubo), was analyzed to provide input data for the Velociraptor claw FE model (FEM). The resultant FEM confirms that dromaeosaurid claws were well-adapted for climbing as they would have been resistant to forces acting in a single (longitudinal) plane, in this case due to gravity. However, the strength of the unguals was limited with respect to forces acting tangential to the long-axis of the claw. The tip of the claw functioned as the puncturing and gripping element of the structure, whereas the expanded proximal portion transferred the load stress through the trabeculae and cortical bone. Enhanced climbing abilities of dromaeosaurid dinosaurs supports a scansorial phase in the evolution of flight.

Claw development and cornification in the passeraceous bird zebrafinch (Taeniatopygia guttata castanotis).

The histogenesis and cornification of claws in zebrafinch embryos has been analyzed. At 10-12 days post-deposition, the epidermis at the tip of the toes forms placode-like anlage associated with a mesenchymal condensation and with a terminal phalange. Claws seem to be modified scales, the dorsal side of which becomes the unguis whereas a ventral scale is the origin of the sub-unguis. At 14-15 days, numerous keratinocytes form the unguis, the corneous layer of which becomes thicker than in the sub-unguis and accumulates beta-keratin and lipids. Keratin bundles are mainly directed toward the tip of the claw and have a prevalent parallel orientation. Unguis corneocytes are thicker and accumulate more beta-keratin than corneocytes of the sub-unguis. Mature corneocytes become partially fused in a compact corneous layer at 17-18 days, near hatching. During growth of the unguis, the embryonic epidermis and beta-keratin cells curve over the tip of the claw and localize in the ventral part of the claw, forming the claw pad. The latter is shed at hatching leaving the pointed claw made of harder corneous layers in the unguis side of the claw.

Immunocytochemistry and protein analysis suggest that reptilian claws contain small high cysteine-glycine proteins.

The present study analyzes the structure and the main proteins of reptilian claws. Mature claws are formed by two to four layers of keratinocytes, a transitional layer of spindle-shaped cells and a thick corneous layer. Transitional cells elongate and merge into a compact corneous layer that is immunoreactive for beta-keratins, now indicated as sauropsid keratin-associated proteins (sKAPs). Most proteins extracted from claws in representative reptiles have a molecular weight of 13-20kDa, an acidic to basic isoelectric point, and are identified from the positive immunoreactivity to beta-keratin antibodies. The comparative analysis between lizard and avian claw beta-keratins shows the presence of an internal region of 20 amino acids with the highest identity, indicated as core-box, within an extended 32-amino acid region with a prevalent beta-sheet secondary conformation. This region is structurally equivalent to a 32-amino acid region present in scale beta-keratins of most reptiles. Both reptilian and avian keratins contain glycine-rich regions for stabilization of the beta-keratin polymer. The N- and C-regions contain most cysteine for disulphide-bonds formation. Claw proteins contain higher amount of cysteine and glycine than other scale proteins, suggesting that claw proteins are specialized cysteine-glycine-rich proteins suited to produce a very hard corneous material.

Equine laminitis: ultrastructural lesions detected 24-30 hours after induction with oligofructose.

REASONS FOR PERFORMING STUDY: The pathology of equine laminitis has been well-documented 48 h after dosing with oligofructose when clinical lameness and lamellar disintegration is well advanced. Further analysis of the earliest lesions, by collecting lamellar samples at the first sign of foot lameness after oligofructose dosing is required in order to increase understanding of the disease. OBJECTIVES: To investigate lamellar epidermal hemidesmosome damage and basement membrane dysadhesion by transmission electron microscopy (TEM). METHODS: Eight clinically normal, mature Standardbred horses were divided randomly into 2 groups of 4. The treatment group were dosed with oligofructose (10 g/kg bwt) and subjected to euthanasia when shifting weight from one foot to other commenced and at the first sign of lameness during walking and turning. This occurred at 24 h in 3 horses and 30 h in one. The sham treatment control group were dosed with water and subjected to euthanasia after 48 h. Lamellar tissues of the front feet were harvested and processed for ultrastructural study using TEM. RESULTS: Examination by TEM showed excessive waviness of the basement membrane zone and pointed tips of some secondary epidermal lamellae, an ultrastructural lesion typical of laminitis. The average number of hemidesmosomes/microm of basement membrane was decreased and their distance from the centre of the lamina densa of the basement membrane was increased. CONCLUSIONS: Laminitis lesions are detectable 24 h after oligofructose administration. POTENTIAL RELEVANCE: Hindgut events occurring in the first 24 h after dosing have begun the destruction of the hoof lamellar interface. Prevention and treatment strategies should precede lameness if they are to be efficacious.

Magnetic resonance microscopy of the equine hoof wall: a study of resolution and potential.

REASONS FOR PERFORMING STUDY: Obtaining magnetic resonance images of the inner hoof wall tissue at the microscopic level would enable early accurate diagnosis of laminitis and therefore more effective therapy. OBJECTIVES: To optimise magnetic resonance imaging (MRI) parameters in order to obtain the highest possible resolution of the structures beneath the equine hoof wall. METHODS: Magnetic resonance microscopy (MRM) was performed in front feet from 6 cadaver horses using T2-weighted fast spin echo (FSE-T2), and T1-weighted gradient echo (GRE-T1) sequences. RESULTS: In T2 weighted FSE images most of the stratum medium showed no signal, however the coronary, terminal and sole papillae were visible. The stratum lamellatum was clearly visible and primary epidermal lamellae could be differentiated from dermal lamellae. CONCLUSION: Most structures beneath the hoof wall were differentiated. Conventional scanners for diagnostic MRI in horses are low or high field. However this study used ultra-high field scanners currently not available for clinical use. Signal-to-noise ratio (S/N) increases as a function of field strength. An increase of spatial resolution of the image results in a decreased S/N. S/N can also be improved with better coils and the resolution of high field MRI scanners will increase as technology develops and surface array coils become more readily available. POTENTIAL RELEVANCE: Although MR images with microscopic resolution were obtained ex vivo, this study demonstrates the potential for detection of lamellar pathology as it occurs. Early recognition of the development of laminitis to instigate effective therapy at an earlier stage and may improve the outcome for laminitic horses. Clinical MR is now readily available at 3 T, while 4 T, 7 T and 9 T systems are being used for human whole body applications.

Membrane-coating granules and the intercellular cementing substance (membrane-coating material) in the epidermis in different regions of the equine hoof.

This study aimed to carry out a comprehensive morphological examination of the structure and ultrastructure of the membrane-coating granules (MCG) and membrane-coating material (MCM) common to all hoof segments as well as segment-specific features in the MCG and MCM regarding their synthesis, structure and biochemical composition. Tissue samples from the hooves of the left front and right hind limb of 20 adult riding horses were used. Samples were preserved and embedded for histological routine methods as well as for histochemical techniques and tracer techniques for light and electron microscopy. The MCM shows a segment-specific structure, biochemical composition and function, which were demonstrated for the different parts of the hoof. The MCM and its properties are related to the segment-specific horn quality. Besides intercellular structural factors the MCM is of major importance for the structural integrity and the biomechanical properties of the hoof horn. The physiological structure and composition of MCM provides explanation for the existence of clinically well-known sites of predilection for hoof disorders such as the White Line Disease. Alterations of the MCM play a role in dysfunction and diseases of the hoof epidermis; they are particularly involved in the pathogenesis of infectious diseases of the hoof.