Norimitsu Saito

The bulge area is the major hair follicle source of nestin-expressing pluripotent stem cells which can repair the spinal cord compared to the dermal papilla

Nestin has been shown to be expressed in the hair follicle, both in the bulge area (BA) as well as the dermal papilla (DP). Nestin-expressing stem cells of both the BA and DP have been previously shown to be pluripotent and be able to form neurons and other non-follicle cell types. The nestin-expressing pluripotent stem cells from the DP have been termed skin precursor or SKP cells. The objective of the present study was to determine the major source of nestin-expressing pluripotent stem cells in the hair follicle and to compare the ability of the nestin-expressing pluripotent stem cells from the BA and DP to repair spinal cord injury. Transgenic mice in which the nestin promoter drives GFP (ND-GFP) were used in order to observe nestin expression in the BA and DP. Nestin-expressing DP cells were found in early and middle anagen. The BA had nestin expression throughout the hair cycle and to a greater extent than the DP. The cells from both regions had very long processes extending from them as shown by two-photon confocal microscopy. Nestin-expressing stem cells from both areas differentiated into neuronal cells at high frequency in vitro. Both nestin-expressing DP and BA cells differentiated into neuronal and glial cells after transplantation to the injured spinal cord and enhanced injury repair and locomotor recovery within four weeks. Nestin-expressing pluripotent stem cells from both the BA and DP have potential for spinal cord regeneration, with the BA being the greater and more constant source.

The role of hair follicle nestin-expressing stem cells during whisker sensory-nerve growth in long-term 3D culture.

We have previously reported that nestin‐expressing hair follicle stem cells can differentiate into neurons, Schwann cells, and other cell types. In the present study, vibrissa hair follicles, including their sensory nerve stump, were excised from transgenic mice in which the nestin promoter drives green fluorescent protein (ND‐GFP mice), and were placed in 3D histoculture supported by Gelfoam®. β‐III tubulin‐positive fibers, consisting of ND‐GFP‐expressing cells, extended up to 500 µm from the whisker nerve stump in histoculture. The growing fibers had growth cones on their tips expressing F‐actin. These findings indicate that β‐III tubulin‐positive fibers elongating from the whisker follicle sensory nerve stump were growing axons. The growing whisker sensory nerve was highly enriched in ND‐GFP cells which appeared to play a major role in its elongation and interaction with other nerves in 3D culture, including the sciatic nerve, the trigeminal nerve, and the trigeminal nerve ganglion. The results of the present report suggest a major function of the nestin‐expressing stem cells in the hair follicle is for growth of the follicle sensory nerve. J. Cell. Biochem. 114: 1674–1684, 2013.

High efficiency genetic modification of hair follicles and growing hair shafts

A technique for genetic modification of hair follicles was developed which results in efficient alteration of the hair shaft phenotype. High-level in vivo transgene expression was maintained in hair follicles such that growing hair shafts were phenotypically altered. Mouse anagen skin fragments, maintained in histoculture, were genetically modified at high efficiency with adenoviral-GFP. The histocultured skin fragments were treated with collagenase which made hair follicles accessible to the adenoviral GFP gene, allowing high-efficiency transduction. These skin fragments were subsequently grafted on to nude mice where GFP was readily visualized in as many as 75% of hair follicles. Most follicles produced GFP-fluorescent growing hair shafts. This technique has produced efficient genetic modification of the hair shaft.

Guidelines for the management of androgenetic alopecia (2010)

Guidelines for the management of androgenetic alopecia (2010) Ryoji TSUBOI, Satoshi ITAMI, Shigeki INUI, Rie UEKI, Kensei KATSUOKA, Sotaro KURATA, Takeshi KONO, Norimitsu SAITO, Motomu MANABE, Masashi YAMAZAKI, GUIDELINES PLANNING COMMITTEE FOR THE MANAGEMENT OF ANDROGENETIC ALOPECIA Department of Dermatology, Tokyo Medical University, Tokyo, Department of Regenerative Dermatology, Osaka University Graduate School of Medicine, Osaka, Department of Dermatology, Juntendo Tokyo Koto Geriatric Medical Center, Tokyo, Department of Dermatology, School of Medicine, Kitasato University, Sagamihara, Kurata Clinic, Beppu, Department of Dermatology, Nippon Medical School, Tokyo, and Department of Dermatology, Akita University Graduate School of Medicine, Akita, Japan

Case of Linear IgA Bullous Dermatosis-involved Ulcerative Colitis

To the Editor: There are a number of reports of linear IgA bullous dermatosis (LABD) associated with preexisting inflammatory bowel disease (IBD), in particular ulcerative colitis (UC).1 We report the case of a patient who developed LABD while receiving treatment for UC. A 28year-old female. UC was diagnosed 1 year ago by colonoscopy (Fig. 1a) and biopsy specimen (Fig. 1b). She was receiving oral mesalazine at 2250 mg daily and prednisolone at 5 mg daily, and her disease activity was well controlled. However, within a few days after she developed a disease relapse, with severe diarrhea and bloody stool, the patient also developed bullae with pruritus on her trunk, gluteal region, and axillary fossa. She was therefore referred to our department. She had noticed clear and tender vesicles and bullae on erosion not associated with erythematous plaques on her trunk and limbs (Fig. 1c). There were no mucosal lesions. Routine laboratory examinations revealed evidence of an acute inflammatory reaction (white blood cell count, 11,000/ L, serum and an elevated erythrocyte sedimentation rate [ESR] of 77 mm/h). An immunological study was performed, including serological tests to determine the presence of autoantibodies, complement levels, rheumatologic markers and immunoglobulin levels, and thyroid tests; however, no significant abnormalities were detected. The skin biopsy specimen showed subepidermal neutrophilrich bullae. Direct immunofluorescence studies showed linear deposition of only IgA on the basement membrane. An indirect immunofluorescence test revealed the presence of circulating anti-basement membrane zone IgA antibodies at a titer of 1:80. Based on these studies, we diagnosed her disease as LABD. She did not receive systemic medications such as vancomycin with subsequent bowel flares, which are sometimes associated with drug-induced linear IgA disease. The intestinal disease activity entered remission spontaneously without any treatments and then, within a few weeks, the eruptions also cleared. No further therapies were required. The patient presented with several episodes of the bullae appearing synchronously with deterioration of the gastrointestinal symptoms and then disappearing within a short time after improvement of the gastrointestinal symptoms. We considered it important to evaluate the pattern of appearance of the skin lesions in relation to the activity of UC as assessed objectively. These usually incorporate the frequency of bowel movements and rectal bleeding, as well as the serum hemoglobin, albumin, and ESR values. The present study used the UC Activity Index (UCAI).2 In our case, the bullae cleared when the UCAI values were below 180, and relapsed when the values exceeded 190 (Fig. 2). Cutaneous diseases have been reported in 10% of patients with UC. Nonspecific eruptions are seen, including urticaria, angioedema, erythema, and purpura.3 In a study of 70 LABD patients selected from a British population, 5 (7.1%) had UC, even though the prevalence of UC in the UK in the general population is only 0.05%, while very few case reports with dermatitis herpetiformis (DH) and UC have been published in the literature.1,4,5 The exact reason for the association between UC and LABD remains unclear. IBDs may induce nonspecific immunoglobulin activation, triggering IgA crossreactive idiotype production against dermoepidermal or epidermal antigens, to produce specific bullous disorders.4,5 Antibodies enter the circulation and are deposited in the skin and mucous membranes because they crossreact with specific peptides in the lamina lucida and sublamina densa regions of the basement membrane. Antibody deposition is thought to stimulate an inflammatory response that damages the basement membrane, leading to mucocutaneous disease.6 Using previously described parameters of activity, we determined the UCAI3 in our patient; the bullae appeared with an increase of the UCAI, and disappeared with a decrease of the UCAI. These findings suggest that the appearance of LABD was related to the activity of UC, with antibodies being produced as the UC activity increased. To the best of our knowledge, this is the first case in which a clear relationship has been documented between LABD and the activity of UC.

Kikuchi-Fujimoto disease developed into autoimmune disease : a report of two cases

We report herein the pathological findings and clinical courses of two cases of Kikuchi–Fujimoto disease (KFD) that developed into autoimmune diseases. The patients are currently undergoing treatment for a disease similar to Sjogren’s syndrome and systemic lupus erythematosus/mixed connective tissue disease. KFD is not an independent condition and most likely develops due to an autoimmune mechanism. Pediatricians should pay careful attention to KFD and encourage long-term follow-up in patients with this condition.

Expression of bleomycin hydrolase in keratinization disorders

A neutral cysteine protease, bleomycin hydrolase (BH), is widely expressed in mammalian tissues, with the skin seeming to contain the highest level. Our previous study revealed that BH transcription is modulated both during differentiation and by cytokines. However, BH involvement in keratinization disorder is not well known. In the present study, we performed immunohistochemical studies of BH and other serine/cysteine proteases in human normal skin and lesional skin with keratinization disorders. BH-positive cells were detected in granular layers of orthokeratotic and hyperkeratotic skin diseases, such as erythrokeratoderma and lichen planus. In parakeratotic skin diseases with porokeratosis, pityriasis rubra pilaris and psoriasis, BH staining was decreased in lesional skins compared to that in normal skin. Similar results were obtained for cysteine proteases, caspase-14 and calpain I. On the other hand, cells positive for serine proteases kallikrein 5 and 7 were increased in parakeratotic and inflammatory skin diseases, such as psoriasis. Semi-quantification analysis revealed that BH- and caspase-14-positive staining had higher intensity than those of the other proteases in normal epidermis. As BH is the major citrulline aminopeptidase in normal granular layer, the alternation would have a significant effect on terminal differentiation processes, such as aberrant processing of deiminated peptides. Therefore, BH may play an important role during the late stage of epidermal differentiation.

Hair follicle stem cell marker nestin expression in regenerating hair follicles of patients with alopecia areata

Cells that are nestin positive and keratin 15 (K15) negative are located in the hair follicle pluripotent stem cell (hfPS) area (hfPSA). The hfPSA is located within the root of the sebaceous glands, in a region just above the hair follicle bulge area. In the current study, we investigated the expression pattern of the stem cell marker nestin in the hair follicle cycling of patients with alopecia areata. In the normal human scalp, the majority of hair follicles are in the anagen phase of development. While it is often difficult to identify nestin expression in late anagen phases, nestin-expressing cells are easily identified in proliferating cells located in the hfPSA of the growing early and middle anagen phase hair follicles. In patients exhibiting alopecia areata, the middle anagen hair follicles with growing cells were found to be nestin positive and K15 negative. In contrast, the hair follicles undergoing degradation in alopecia areata patients demonstrated lymphocytic infiltration within the nestin- and K15-negative dermal papilla cells. Both the nestin-positive hfPSA and K15-positive hair follicle bulge areas were not damaged in all phases. In addition, the regenerating early anagen hair follicles demonstrated nestin-positive and K15-negative cells within the dermal papilla and in the area surrounding the hair bulb. These results suggest that the nestin-positive cells play an important role not only in the hfPSA, but also in the dermal papilla in the regenerating hair follicle.

Guidelines for the diagnosis and treatment of male‐pattern and female‐pattern hair loss, 2017 version

Male‐pattern hair loss (MPHL, androgenetic alopecia) is a slowly progressive form of alopecia which begins after puberty. In 2010, we published the first Japanese edition of guidelines for the diagnosis and treatment of MPHL. It achieved the original goal of providing physicians and patients in Japan with evidence‐based information for choosing efficacious and safe therapy for MPHL. Subsequently, new therapeutic drugs and treatment methods have been developed, and womens perception of MPHL has undergone change and the term “female‐pattern hair loss (FPHL)” is becoming more common internationally. Thus, here we report a revised version of the 2010 guidelines aimed at both MPHL and FPHL. In these guidelines, finasteride 1 mg daily, dutasteride 0.5 mg daily and topical 5% minoxidil twice daily for MPHL, and topical 1% minoxidil twice daily for FPHL, are recommended as the first‐line treatments. Self‐hair transplantation, irradiation by light‐emitting diodes and low‐level lasers, and topical application of adenosine for MPHL are recommended, whereas prosthetic hair transplantation and oral administration of minoxidil should not be performed. Oral administration of finasteride or dutasteride are contraindicated for FPHL. In addition, we have evaluated the effectiveness of topical application of carpronium chloride, t‐flavanone, cytopurine, pentadecane and ketoconazole, and wearing a wig. Unapproved topical application of bimatoprost and latanoprost, and emerging hair regeneration treatments have also been addressed. We believe that the revised guidelines will improve further the diagnostic and treatment standards for MPHL add FPHL in Japan.

Nestin-Expressing Hair-Follicle-Associated Pluripotent (HAP) Stem Cells Promote Whisker Sensory-Nerve Growth in Long-Term 3D-Gelfoam® Histoculture.

Mouse whiskers containing hair-follicle-associated pluripotent (HAP) stem cells, from nestin-driven green fluorescent protein (ND-GFP) transgenic mice, were placed in 3D histoculture supported by Gelfoam(®). β-III tubulin-positive fibers, consisting of ND-GFP-expressing HAP stem cells, extended up to 500 mm from the whisker nerve stump in histoculture. The growing fibers had growth cones on their tips expressing F-actin indicating they were growing axons. The growing whisker sensory nerve was highly enriched in ND-GFP HAP stem cells which appeared to play a major role in its elongation and interaction with other nerves placed in 3D culture, including the sciatic nerve, the trigeminal nerve, and the trigeminal nerve ganglion. The results suggested that a major function of HAP stem cells in the hair follicle is for growth of the hair follicle sensory nerve.