James A. Vernon

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.

Characterization of the exocuticle a-layer proteins of wool.

Abstract:  The outermost protein layer of wool cuticle cells is known as the exocuticle a‐layer. This layer is a resistant barrier to the degradation of the fibre and, as a result, little is known of its proteinaceous composition. Merino wool fibres were subjected to both proteolytic and chemical digestion and the resulting material was found by transmission electron microscopy to be highly enriched in a‐layer. Amino acid analysis revealed a high cysteine and glycine content, with a close, but not exact, match to the Allwörden membrane. Subsequent digestion of the a‐layer preparation by 2‐nitro‐5‐thiocyano‐benzoic acid produced a large number of short peptides, and analysis by mass spectrometry revealed peptides with strong homologies to cuticular ultra‐high sulphur proteins of sheep wool and cuticular ultra‐high and high‐sulphur proteins of human hair, thus supporting other evidence for the presence of these sulphur‐rich proteins in the a‐layer.

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.

Redox proteomic evaluation of bleaching and alkali damage in human hair

Protein modification and damage in human hair, resulting from environmental, cosmetic and grooming stresses, create changes to visual and tactile characteristics and correlates with consumer perception of quality. This study outlines molecular‐level evaluation of modification resulting from peroxide (bleaching) and alkaline straightening (relaxing) treatments.

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.