Joy L. Woods

Cortical cell types and intermediate filament arrangements correlate with fiber curvature in Japanese human hair

Naturally straight and curved human scalp hairs were examined using fluorescence and electron microscopy techniques to determine morphological and ultrastructural features contributing to single fiber curvature. The study excluded cuticle and medulla, which lack known bilateral structural asymmetry and therefore potential to form curved fibers. The cortex contained four classifiable cell types, two of which were always present in much greater abundance than the remaining two types. In straight hair, these cell types were arranged annularly and evenly within the cortex, implying that the averaging of differing structural features would maintain a straight fiber conformation. In curved fibers, the cell types were bilaterally distributed approximately perpendicular to fiber curvature direction with one dominant cell type predominantly located closest to the convex fiber side and the other, closest to the concave side. Electron tomography confirmed that the dominant cell type closest to the convex fiber side contained discrete macrofibrils composed of helically arranged intermediate filaments, while the dominant cell type closest to the concave side contained larger fused macrofibrils composed of intermediate filament arrangements varying from helical to hexagonal arrays approximately parallel to the longitudinal fiber axis. These findings concur with the current hypothesis of hair curvature formation and behavior.

Arrangement of trichokeratin intermediate filaments and matrix in the cortex of Merino wool.

Tomograms of transverse sections of Merino wool fibers obtained from fleeces differing in fiber curvature were reconstructed from image series collected using a 300kV transmission electron microscope. Trichokeratin intermediate filaments (IFs) from the ortho-, para- and mesocortices were modeled from the tomograms. IFs were predominantly arranged in left-handed concentric helices with the relative angle of IFs increasing progressively from the center to the periphery of orthocortex macrofibrils. The median increase in IF angle between adjacent IFs between the center and periphery was 2.5°. The length of one turn of the helical path of an IF was calculated to be approximately 1μm for an IF tilted at 30° and positioned 100nm from the macrofibril center. With the exception of one paracortex macrofibril that weakly resembled an orthocortex macrofibril, all para- and mesocortex macrofibrils modeled had a parallel arrangement of the IFs, with a more ordered arrangement found in the mesocortex. Within the limited sample set, there appeared to be no significant relationship between IF angle and fiber curvature. We examined the matrix/IF ratio (in the form of proportion of matrix to one IF, calculated from IF center-to-center distance and IF diameter) for 28 macrofibrils used for modeling. The proportion of matrix was significantly different in the different cortex cell types, with paracortex having the most (0.61), orthocortex having the least (0.42), and mesocortex being intermediate (0.54). Fibers of different crimp type (high, medium or low crimp) were not significantly different from each other with respect to matrix proportion.

The proteomics of wool fibre morphogenesis.

Gel and gel-free proteomic techniques have been used for the first time to directly study the proteins present in whole wool follicles and dissected portions of follicles that correlated with morphological changes in the developing fibre as determined by transmission electron microscopy. Individual wool follicles were dissected into four portions designated as the bulb, elongation, keratogenous and keratinisation portions. Gel-free proteomic analysis of dissected portions from 30 follicles showed that the first keratins to appear were K31, K35 and K85, in the bulb portion. The first epithelial KAP, trichohyalin, was detected in the bulb portion and the first cortical KAP, KAP11.1 was found in the elongation portion. Other major trichocyte keratins and cortical KAPs began to appear further up the follicle in the keratogenous and keratinisation zones. These results were consistent with what has been observed from gene expression studies and correlated well with the morphological changes observed in the follicle. Other proteins detected by this approach included the keratin anchor protein desmoplakin, as well as vimentin and epithelial keratins, histones, ribosomal proteins and collagens. Two-dimensional electrophoretic (2DE) analysis of dissected portions of 50 follicles revealed substantial changes in the position, number and intensity of the spots of the trichocyte keratins as they progressed through the follicle zones, suggesting that they are subject to modification as a result of the keratinisation process. Also present in the 2DE maps were a number of epithelial keratins, presumably from the inner and outer root sheaths, and the dermal components.

Transmission electron microscopy staining methods for the cortex of human hair: a modified osmium method and comparison with other stains

For wool, superior staining of a wide range of ultrastructural components is achieved by en bloc treatment of fibres with a chemical reductant followed by osmium tetroxide. For human scalp hair, although staining quality is similar, the penetration of reagents is poor, resulting in large parts of the fibre cortex remaining unstained. Here we describe a modification to the reduction–osmication method in which reagents penetrate through a cut fibre end, allowing visualization of a wide range of features across the cortex. We compare the staining quality, artefacts and range of structure rendered visible using transmission electron microscopy for en bloc reduction–osmication to other staining alternatives including en bloc silver nitrate and section stains based on uranyl acetate and lead citrate, phosphotungstic acid, potassium permanganate, ammoniacal silver nitrate and some combinations of these stains. The effects of hair‐care treatments are briefly examined.

Three-dimensional architecture of macrofibrils in the human scalp hair cortex.

Human scalp hairs are comprised of a central cortex enveloped by plate-like cuticle cells. The elongate cortex cells of mature fibres are composed primarily of macrofibrils-bundles of hard-keratin intermediate filaments (IFs) chemically cross-linked within a globular protein matrix. In wool, three cell types (ortho-, meso- and paracortex) contain macrofibrils with distinctly different filament arrangements and matrix fractions, but in human hair macrofibril-cell type relationships are less clear. Here we show that hair macrofibrils all have a similar matrix fraction (∼0.4) and are typically composed of a double-twist architecture in which a central IF is surrounded by concentric rings of tangentially-angled IFs. The defining parameter is the incremental angle increase (IF-increment) between IFs of successive rings. Unlike the wool orthocortex, hair double-twist macrofibrils have considerable inter-macrofibril variation in IF increment (0.05-0.35°/nm), and macrofibril size and IF increment are negatively correlated. Correspondingly, angular difference between central and outer-most IFs is up to 40° in small macrofibrils, but only 5-10° in large macrofibrils. Single cells were observed containing mixtures of macrofibrils with different diameters. These new observations advance our understanding of the nano-level and cell-level organisation of human hair, with implications for interpretation of structure with respect the potential roles of cortex cell types in defining the mechanical properties of hair.

Morphology and ultrastructure of antler velvet hair and body hair from red deer (Cervus elaphus)

We provide a detailed description of the ultrastructure of deer hair fibers. Guard hairs and underhairs from the winter coat of red deer (Cervus elaphus), and antler velvet hairs from the same species were examined. All fibers displayed the typical keratin fiber morphology of overlapping cuticle cells surrounding a core of cortex cells, and often a centrally‐located medulla, but there were considerable differences in the diameter, cuticle thickness, and scale pattern, and in the relative amounts of cortex and medulla along individual fibers, and between the different types of fiber. In addition, closer examination of cortex cells using transmission electron microscopy revealed considerable differences in the arrangement of intermediate filaments in the different fiber types. Fine underhairs appeared similar to fine wool fibers from sheep because intermediate filament arrangements were very similar to those found in wool orthocortex cells and paracortex cells. In addition, a similar bilateral distribution of these cell types was evident. However, in the antler velvet hairs and the guard hairs, intermediate filament arrangements were more variable and complex, and showed similarities to those in heterotype cortex cells described for human hair. J. Morphol., 2011.

Intrinsic curvature in wool fibres is determined by the relative length of orthocortical and paracortical cells

ABSTRACT Hair curvature underpins structural diversity and function in mammalian coats, but what causes curl in keratin hair fibres? To obtain structural data to determine one aspect of this question, we used confocal microscopy to provide in situ measurements of the two cell types that make up the cortex of merino wool fibres, which was chosen as a well-characterised model system representative of narrow diameter hairs, such as underhairs. We measured orthocortical and paracortical cross-sectional areas, and cortical cell lengths, within individual fibre snippets of defined uniplanar curvature. This allowed a direct test of two long-standing theories of the mechanism of curvature in hairs. We found evidence contradicting the theory that curvature results from there being more cells on the side of the fibre closest to the outside, or convex edge, of curvature. In all cases, the orthocortical cells close to the outside of curvature were longer than paracortical cells close to the inside of the curvature, which supports the theory that curvature is underpinned by differences in cell type length. However, the latter theory also implies that, for all fibres, curvature should correlate with the proportions of orthocortical and paracortical cells, and we found no evidence for this. In merino wool, it appears that the absolute length of cells of each type and proportion of cells varies from fibre to fibre, and only the difference between the length of the two cell types is important. Implications for curvature in higher diameter hairs, such as guard hairs and those on the human scalp, are discussed. Highlighted Article: Curvature in mammalian hairs is underpinned by the relative difference in length between orthocortical and paracortical cells rather than their proportion or number along each side of the fibre.

Like Follicle, like Fibre? Diameter and not Follicle Type Correlates with Fibre Ultrastructure

The hair follicles of most mammals are of two types, primary and secondary. Primary follicles develop earlier and have a prominent arrectorpili muscle. Secondary follicles have less prominent muscles and are often clumped, sharing a common opening from which fibres emerge. It is not entirely clear what types of follicles occur in human scalps. Partly this is because human hairs have a uniform appearance, unlike many mammals in which robust primary hairs differ markedly from narrow secondary fibres. Some sheep breeds are an exception because like humans, wool fibres have a similar macro-scale appearance irrespective of follicle type. How deep does this similarity go Using electron microscopy, we examined wool primary fibres from different breeds and contrasted them to secondary fibres. For fibres of similar diameter, there was no significant difference in the ultrastructure or proportion and distribution of cortex cell types in primary and secondary fibres. We conclude that fibre diameter is the most important fibre parameter with respect to structural differences between fibres, not whether the fibres originate from primary or secondary follicles.

The physical and chemical disruption of human hair after bleaching - studies by transmission electron microscopy and redox proteomics

To understand the structural and chemical effects of cosmetic peroxide bleaching on human hair.

The proteomics of wool follicles

Messenger RNA hybridisation labelling of hair follicles has revealed that in both sheep and humans there is a sequential pattern to expression of the major keratin and keratin associated proteins (Langbein and Schweizer, 2005; Yu et al., 2009). How expression sequence and post-translational changes relate to fibre development is largely unknown. Thus, this study was initiated to determine if proteomics could provide an insight into protein changes during the various stages of protein synthesis and keratinisation in the maturing wool fibre.