Three-dimensional correlative light and focused ion beam scanning electron microscopy reveals the distribution and ultrastructure of lanceolate nerve endings surrounding terminal hair follicles in human scalp skin.

Lanceolate nerve endings (LNEs) surrounding hair follicles (HFs) play an important role in detecting hair deflection. Complexes of the LNEs form a palisade-like structure along the longitudinal axis of hair roots in which axons are sandwiched between two processes of terminal Schwann cells (tSCs) at the isthmus of HFs. The structure and molecular mechanism of LNEs in animal sinus hair, pelage, and human vellus hairs have been investigated. Despite the high density of HFs in human scalp skin, the LNEs in human terminal HFs have not been investigated. In this study, we aimed to reveal the distribution and ultrastructure of LNEs in terminal HFs of human scalp skin. Using light-sheet microscopy and immunostaining, the LNEs were observed at one terminal HF but not at the other terminal HFs in the same follicular unit. The ultrastructure of the LNEs of terminal HFs in human scalp skin was characterized using correlated light and electron microscopy (CLEM). Confocal laser microscopy and transmission electron microscopy of serial transverse sections of HFs revealed that LNEs were aligned adjacent to the basal lamina outside the outer root sheath (ORS), at the isthmus of terminal HFs, and adjacent to CD200-positive ORS cells in the upper bulge region. Moreover, axons with abundant mitochondria were sandwiched between tSCs. Three-dimensional CLEM, specifically confocal laser microscopy and focused ion beam scanning electron microscopy, of stained serial transverse sections revealed that LNEs were wrapped with type I and type II tSCs, with the processes protruding from the space between the Schwann cells. Moreover, the ultrastructures of LNEs at miniaturized HFs were similar to those of LNEs at terminal HFs. Preembedding immunoelectron microscopy revealed that Piezo-type mechanosensitive ion channel component 2 (Piezo2), a gated ion channel, was in axons and tSCs and adjacent to the cell membrane of axons and tSCs, suggesting that LNEs function as mechanosensors. The number of LNEs increased as the diameter of the ORS decreased, suggesting that LNEs dynamically adapt to the HF environment as terminal HFs miniaturize into vellus-like hair. These findings will provide insights for investigations of mechanosensory organs, aging, and re-innervation during wound healing.

Hair follicle dystrophy in a litter of domestic cats resembling lanceolate hair mutant mice.

BACKGROUND: A new congenital hair-shaft abnormality resembling the lanceolate hair phenotype of rodents is described in a litter of four domestic short hair (DSH) cats. Data relating to hair shaft and follicle disorders remain scarce in veterinary medicine. OBJECTIVES: To describe and compare structural abnormalities in these cats with other hair dystrophies in cats and other mammals. ANIMALS: A DSH cat litter with progressive noninflammatory alopecia. METHODS AND MATERIALS: Histopathological evaluation, scanning and transmission electron microscopy, and X-ray based element analysis defined the hair and skin changes in cats born with alopecia. Findings were compared to archival data from normal cats and lanceolate hair (Dsg4lahJ ) and Keratin 75 (Krt75tm1Der ) mutant mice. RESULTS: Light and scanning electron microscopy of the hairs revealed lance- or spear-head shaped defects of the hair tip. Histological findings were swollen hair shafts, initially above the hair bulb matrix and later found in the distal parts of the telogen hair follicles, similar to those observed in Dsg4lahJ Krt75tm1Der mutant mice. Transmission electron microscopy of the hair shaft and hair follicles showed a loss in the normal structure of the guard hairs in the alopecic cats. There was a statistically significant decrease in sulfur content just below the defects in the hair shafts (trichothiodystrophy). CONCLUSION AND CLINICAL IMPORTANCE: A rare form of congenital alopecia resulting in follicular dystrophy is described in cats which is similar to hair follicle and hair-shaft changes reported in several mutant mouse strains with single gene mutations in adhesion molecules or keratin genes.

Quantifying the impact of braiding and combing on the integrity of natural African hair.

BACKGROUND: Afro hair breakage is most commonly attributed to grooming practices such as braiding, hair extensions and weaves, both for chemically treated hair and natural Afro hair. These grooming practices are also frequently associated with traction alopecia in clinical studies. However, there is little to no quantitative scientific data on the surface and internal hair fibre damage caused by grooming, including combing, on Afro hair. METHODS: Qualitative and quantitative techniques such as scanning electron microscopy, cuticle cohesion and tensile testing were used to determine the effects of frequent braiding on the integrity of female Afro hair in 15 frequent braiders and 15 occasional braiders (control group) in Johannesburg, South Africa. Frequent braiders were women who braided more than 8 times per year, while occasional braiders typically limit braiding to only twice 2 per year. The severity of traction alopecia was quantified in both groups using the standard Marginal Traction Alopecia Scoring system. Surface damage on naturally curly Afro hair, that was combed but not braided, was assessed as another hairstyle option. RESULTS: All measurements showed a relationship between surface and internal hair fibre damage and braiding frequency. Frequent braiders had thinner hair, with more cuticle damage as shown by SEM and cuticle cohesion assessments. Their hair fibres were also significantly weaker (p < 0.05), as shown by the tensile testing. In addition, they also had higher traction alopecia severity scores. In the combing impact assessment, a linear trend was observed between surface hair fibre damage and the number of combing strokes. Cuticle damage from 480 combing strokes was more severe than that from frequent braiding. CONCLUSION: Frequent braiding and combing damage natural Afro hair. Quantification of damage caused by different grooming practices identifies a need to develop products that could help mitigate specific hair damage associated with grooming practices. The degree of damage and subsequent hair loss could also be minimized by education on best braiding practices to protect the hair fibre and avoid hair follicle miniaturization caused by high traction hairstyles.

CONTEXTE: La casse des cheveux d'origine Africaine est le plus souvent associées à des pratiques cosmétiques telles que le tressage et les extensions de cheveux, à la fois pour les cheveux traités chimiquement et les cheveux naturels. Ces pratiques cosmétiques sont également fréquemment associées à une alopécie de traction, comme le relatent les études cliniques. Cependant, il existe peu ou pas de données scientifiques quantitatives sur l'impact de ce type de pratiques, y compris le peignage, sur les dommages internes et de surface des cheveux d'origine africaine. MÉTHODES: Des techniques qualitatives et quantitatives, comme la microscopie électronique à balayage (MEB), la cohésion cuticulaire et des tests de traction, ont été utilisés pour déterminer les effets du tressage sur l'intégrité des cheveux de deux groupes de 15 femmes Africaines (à Johannesburg, Afrique du Sud) : un premier groupe de femmes pratiquant un tressage fréquent et un autre groupe un tressage occasionnel. Le groupe des tresseurs fréquents correspond à des femmes qui pratiquent le tressage plus de 8 fois par an, tandis que les tresseurs occasionnels limitent généralement le tressage à seulement deux fois par an. La gravité de l'alopécie de traction a été également quantifiée dans les deux groupes à l'aide du système standard de notation de l'alopécie tractionnelle. Parallèlement, à titre de comparaison, les dommages de surface ont été également évalués sur des cheveux d'origine Africaine naturellement bouclés, non tressés, mais qui ont subi un peignage répété. RÉSULTATS: Toutes les mesures ont montré une relation entre les dommages de surface et internes de la fibre et la fréquence de tressage. Les tresseurs fréquents ont des cheveux plus fins, avec un endommagement plus important de la cuticule, comme le montrent les évaluations MEB et la cohésion cuticulaire. Les cheveux sont également significativement plus fragiles (p <0,05), comme le montrent les tests de traction. De plus, ces femmes ont également des scores d'alopécie de traction plus élevés. Dans le cas spécifique du peignage répété, une tendance linéaire a été observée entre les dommages en surface de la fibre capillaire et le nombre de coups de peigne. On peut également noter que l'endommagement de la cuticule causé par 480 coups de peigne est plus important que celui causé par des tressages fréquents. CONCLUSION: Les tressages et peignages fréquents endommagent les cheveux naturels d'origine Africaine. La quantification des dommages causés par différentes pratiques cosmétiques met en évidence le besoin de développer des produits qui pourraient aider à atténuer les dommages liés à des pratiques spécifiques. Le niveau de dommage et la perte de cheveux pourraient également être minimisés par un apprentissage de meilleures pratiques de tressage pour protéger la fibre et éviter la diminution des follicules pileux causée par des coiffures qui entrainent un forte traction sur la fibre.

High-pressure freezing followed by freeze substitution of a complex and variable density miniorgan: the wool follicle.

Cryofixation by high-pressure freezing (HPF) followed by freeze substitution (FS) is a preferred method to prepare biological specimens for ultrastructural studies. It has been shown to achieve uniform vitrification and ultrastructure preservation of complex structures in different cell types. One limitation of HPF is the small sample volume of <200 µm thickness and about 2000 µm across. A wool follicle is a rare intact organ in a single sample about 200 µm thick. Within each follicle, specialized cells derived from multiple cell lineages assemble, mature and cornify to make a wool fibre, which contains 95% keratin and associated proteins. In addition to their complex structure, large density changes occur during wool fibre development. Limited water movement and accessibility of fixatives are some issues that negatively affect the preservation of the follicle ultrastructure via conventional chemical processing. Here, we show that HPF-FS of wool follicles can yield high-quality tissue preservation for ultrastructural studies using transmission electron microscopy.

The walking dead: sequential nuclear and organelle destruction during hair development.

BACKGROUND: Transition of hair shaft keratinocytes from actively respiring, nucleated cells to structural cells devoid of nucleus and cytoplasm is key to hair production. This form of cell 'death', or cornification, requires cellular organelle removal to allow the cytoplasm to become packed with keratin filament bundles that further require cross-linking to create a strong hair fibre. Although these processes are well described in epidermal keratinocytes, there is a lack of understanding of such mechanisms, specifically in the hair follicle. OBJECTIVES: To gain insights into cornification mechanisms within the hair follicle and thus improve our understanding of normal hair physiology. METHODS: Scalp biopsies and hair-pluck samples were obtained from healthy human donors and analysed microscopically after immunohistochemical staining. RESULTS: A focal point of respiratory activity was evident in keratogenous zone cells within the hair shaft, which also exhibited nuclear damage. Nuclear degradation occurred via both caspase-dependent and caspase-independent pathways. Conversely, mitophagy was driven by Bnip3L and restricted to the boundary of the keratogenous zone at Adamson's Fringe. CONCLUSIONS: We propose a model of stepwise living-dead transition within the first 1 mm of hair formation, whereby fully functional, nucleated cells first consolidate required functions by degrading nuclear DNA, yet continue to respire and provide the source of reactive oxygen species required for keratin cross-linking. Finally, as the cells become packed with keratin bundles, Bnip3L expression triggers mitophagy to rid the cells of the last remaining 'living' characteristic, thus completing the march from 'living' to 'dead' within the hair follicle.

Comparing dermoscopy and histological examination of normal equine skin.

BACKGROUND: Dermoscopy is a noninvasive diagnostic technique that allows visualization of structures of the superficial dermis not visible with the naked eye. HYPOTHESIS/OBJECTIVES: To assess the usefulness and applicability of dermoscopy for evaluation of healthy equine skin. ANIMALS: Twelve healthy horses from a research herd. METHODS: Five regions (cheek, lateral neck, dorsum, flank and abdomen) were examined with contact dermoscopy using both nonpolarized and polarized light at both 17-fold and 24-fold magnification. These findings were compared to histological features of skin biopsies cut both longitudinally and transversely. RESULTS: Using a hand-held dermatoscope with nonpolarized light, epidermal ridges were observed. Using polarized light, follicular openings and distinctly separate epidermal openings of sweat gland ducts were observed in some but not all individuals. Similarities were noted between histological and dermoscopic results. CONCLUSIONS: Although not ideal for visualizing many structures in the superficial dermis of healthy equine skin, dermoscopy allowed visualization of epidermal ridges, hair shafts in the infundibular portion of the hair follicles and sweat gland duct openings. Dermoscopy could potentially be useful in the evaluation of diseases affecting the sweat glands, epidermis and hair shaft.

Hair Follicle Miniaturization in a Woolly Hair Nevus: A Novel "Root" Perspective for a Mosaic Hair Disorder.

Woolly hair nevus is a mosaic disorder characterized by unruly, tightly curled hair in a circumscribed area of the scalp. This condition may be associated with epidermal nevi. We describe an 11-year-old boy who initially presented with multiple patches of woolly hair and with epidermal nevi on his left cheek and back. He had no nail, teeth, eye, or cardiac abnormalities. Analysis of plucked hairs from patches of woolly hair showed twisting of the hair shaft and an abnormal hair cuticle. Histopathology of a woolly hair patch showed diffuse hair follicle miniaturization with increased vellus hairs.

Alterations in Hair Follicle Morphology and Hair Shaft Production After Follicular Unit Transplantation.

Follicular unit transplantation is the most commonly performed technique in modern restorative hair transplantation surgery. It relies on the acquisition of intact follicular units from microdissected scalp skin strips and their subsequent transplantation into the recipient regions affected by alopecia. Ideally, the translocation of follicular units from the balding-resistant areas of the scalp (usually the occipital region) to the recipient site should not result in any morphological change in the grafts. Nevertheless, the insults associated with surgical intervention present grafted follicles to mechanical and chemical cues differently from those of the physiological steady-state conditions in undamaged skin. This disruption of the normal follicular microenvironment might alter important aspects of hair biology in grafts, for example, hair cycle and pigmentation, and, in turn, could lead to differences in hair appearance, eventually culminating in a diminished esthetical outcome of the surgery. In this study, the authors analyzed native and grafted scalp hair follicles (HFs) from 2 patients who had undergone follicular unit transplantation surgeries formerly. Scanning electron microscopy and light microscopy-based histomorphometry revealed a marked enlargement of follicular structures in the grafts with a concomitant increase in hair shaft diameter. Immunohistological staining confirmed a thickening of the dermal sheath in transplanted HFs that also harbored a denser vascular network. Taken together, these results show that the grafted HFs analyzed were subjected to marked morphological changes during their residence in the recipient site and that this phenomenon is associated with a modulation of follicular vascularization.

Loose Anagen Syndrome: A Retrospective Chart Review of 37 Cases.

BACKGROUND/OBJECTIVES: Loose anagen syndrome (LAS) is a disorder of abnormal anchorage of the hair to the scalp. Its symptoms include an inability to grow hair long and hair that is easily pulled out. METHODS: We conducted a retrospective chart review of patients with LAS over the last 10 years at the Children's Hospital of Philadelphia and found 37 cases. RESULTS: LAS was more common in females and was found in all hair colors and skin types. CONCLUSION: Diagnosis of LAS should be made based on history, microscopic findings, and hair bulb and shaft features.

Follicular necrotic keratinocytes - a helpful clue to the diagnosis of measles.

A skin specimen taken from the exanthem of a 22-year-old man with measles infection was examined by light microscopy. The most characteristic changes were foci of multiple necrotic keratinocytes, partially confluent and partially isolated, strictly within follicles and sebaceous glands. Serial sections revealed occasional necrotic and none-necrotic syncytial-type multinucleated epithelial cells in some infundibula. Neither nuclear nor cytoplasmic viral inclusions were present.

Scanning Electron Microscopic Hair Shaft Analysis in Ectodermal Dysplasia Syndromes.

OBJECTIVES: The objective of the current study was to catalog hair shaft abnormalities in individuals with ectodermal dysplasia (ED) syndromes using scanning electron microscopy (SEM) and to compare the findings with those in unaffected controls. This is the second of a two-part study, the first of which used light microscopy as the modality and was previously published. METHODS: Scanning electron microscopy was performed in a blinded manner on hair shafts from 65 subjects with seven types of ED syndromes and 41 unaffected control subjects. Assessment was performed along the length of the shaft and in cross section. SETTING: Hair donations were collected at the 28th Annual National Family Conference held by the National Foundation for Ectodermal Dysplasia. Control subjects were recruited from a private dermatology practice and an academic children's hospital outpatient dermatology clinic. RESULTS: SEM identified various pathologic hair shaft abnormalities in each type of ED and in control patients. When hairs with all types of ED were grouped together and compared with those of control patients, the difference in the presence of small diameter and shallow and deep grooves was statistically significant (p < 0.05). When the EDs were separated according to subtype, statistically significant findings were also seen. CONCLUSION: SEM is a possible adjuvant tool in the diagnosis of ED syndromes. There are significant differences, with high specificity, between the hairs of individuals with ED and those of control subjects and between subtypes.

Selective hair therapy: bringing science to the fiction.

Investigations on carrier-based drug delivery systems for higher selectivity in hair therapy have clearly evolved from dye release and model studies to highly sophisticated approaches, many of which specifically tackle hair indications and the delivery of hair-relevant molecules. Here, we group recent hair disease-oriented work into efforts towards (i) improved delivery of conventional drugs, (ii) delivery of novel drug classes, for example biomolecules and (iii) targeted delivery on the cellular/molecular level. Considering the solid foundation of experimental work, it does not take a large step outside the current box of thinking to follow the idea of using large carriers (>500 nm, unlikely to penetrate as a whole) for follicular penetration, retention and protection of sensitive compounds. Yet, reports on particles <200 nm being internalized by keratinocytes and dendritic cells at sites of barrier disruption (e.g., hair follicles) combined with recent advances in nanodermatology add interesting new facets to the possibilities carrier technologies could offer, for example, unprecedented levels of selectivity. The authors provide thought-provoking ideas on how smart delivery technologies and advances in our molecular understanding of hair pathophysiology could result in a whole new era of hair therapeutics. As the field still largely remains in preclinical investigation, determined efforts towards production of medical grade material and truly translational work are needed to demonstrate surplus value of carrier systems for clinical applications.

Growth and viability of Liaoning Cashmere goat hair follicles during the annual hair follicle cycle.

Here, we studied hair follicle development of Liaoning Cashmere goats. Every month for 1 year, skin samples were collected from five 1.5-year-old female goats, and made into paraffin sections. A number of parameters were measured of primary and secondary hair follicles via microscopic observation including follicle depth, hair bulb width, dermis and epidermis thickness, changes in follicle activity, and histology. The results showed the presence of three phases in the annual hair cycle: anagen, catagen, and telogen. Primary and secondary hair follicle depth varied across the months; however, no significant difference was obtained between adjacent months (P>0.05). Primary hair follicles had a bigger hair bulb width compared to secondary hair follicles; however, this difference declined during hair follicle developed in anagen. As hair follicle growth slowed, the hair bulb broadened, and hair root depth became shallower. During the entire hair cycle, hair follicle depth and dermis thickness were positively correlated; however, this relationship was not significant (P>0.05) for primary and secondary hair follicle density and the ratio of secondary hair follicle density and primary hair follicle density (S/P ratio). In addition, new and old primary hair follicles coexisted with secondary hair follicles. Finally, secondary hair follicles had a higher activity rate compared to primary hair follicle in adult Liaoning Cashmere goats in certain months.

Evaluation of ATP content in hair bulbs in human scalp.

The content of ATP in scalp hair bulbs in humans was measured in the hair roots from 15 healthy volunteers. Light and electron microscopy confirmed the presence of outer and an inner root sheaths in the root of pulled out anagen hair. Incubation of samples in buffer solution led to extraction of ATP, which was measured by the chemiluminescent method. Mechanic disintegration of hair bulbs and their freezing-defrosting did not increase ATP output. The results of microscopy indicated that ATP extraction procedure was associated with separation of the outer radical sheath from the inner one without impairing the structure of the inner sheath. The mean content of ATP was 12 ± 2 pmol per bulb. The use of pulled out hair bulbs for ATP measurements simplified the procedure as involved no surgical removal of follicles.

[Cellular dynamics of the outer layers of the hair follicle of fine-wool sheep during the phase of stable hair growth].

The structure, origin, and migration of outer sheath cells of the hair follicles of domestic sheep were studied by electron microscopic, autoradiographic, and histochemical (glycogen) in order to understand the role of this layer in hair morphogenesis. We demonstrated that the cells of the outer layers of the outer sheath interpose into the inner "companion" layer of the outer sheath. Although this process takes place all along the hair follicle from the lower bulb up to the sebaceous glands orifices, it mainly takes place over the bulb. Labeled cells interposed into the companion layer move towards sebaceous glands orifices more than 24 hours faster than labeled cells of the inner sheath and hair, because these cells included the label not in the bulb cambium (as hair and inner sheath) but over the bulb, and from this point they start movement. Interposition of cells into the companion layer must cause increase of its volume and additional volume supposed to be led away into the pillar canal around the hair near the sebaceous glands orifices. This can provide the mechanism for the propagation of the hair and inner sheath promotion to sebaceous gland orifices.

Immunolocalization of telomerases in human hairs identifies proliferating cells in the bulb matrix and outer root sheath.

Telomerase is present in cells with numerous or even un-limited replicative cycles, and some studies suggest it is a stemness marker. In order to determine whether this is the case for the human hair bulbs, an immunohistochemical and ultrastructural study has been carried out using antibodies against telomerase and PCNA (a cell proliferation marker). The observed labeling is similar for the two antibodies here utilized and is mainly nuclear. More frequent telomerase-positive cells are seen in the matrix epithelium of anagen hair bulbs but sparse labeled cells are also seen in the outer root sheath. In late catagen and also in telogen hair follicles only sparse labeled cells are present in the outer root sheath and few cells also in the secondary germinal epithelium formed at the base of the hair bulb in telogen. Electron microscopic immunogold shows a prevalent nuclear distribution and a lower cytoplasmic distribution in sparse cells of anagen bulb matrix that contain few keratin bundles. The nuclear localization is generally seen over the euchromatin or in areas occupied by more compact chromatin that may indicate an activity of telomerase in chromatin assemblage or dis-assemblage. The study concludes that the localization of telomerase is present in cells undergoing proliferation, namely transit amplifying cells of the outer root sheath that are sparsely detected in the lowermost secondary germinal hair bulb also in telogen.