Destruction Of Follicles Research Articles

Many Paths to Alopecia via Compromised Regeneration of Hair Follicle Stem Cells

Alopecia can be caused by defective formation, defective regeneration, or increased destruction of hair follicles. Much work has elucidated the roles of diffusible morphogens in modulating hair follicle stem cell activities. Recent studies have revealed novel molecular events within the nucleus, which are required for the activation and progression of hair stem cells. These studies will provide new clues and targets for designing therapeutic strategies for hair loss.

Scrambled and fried: Cigarette smoke exposure causes antral follicle destruction and oocyte dysfunction through oxidative stress

Cigarette smoke is a reproductive hazard associated with pre-mature reproductive senescence and reduced clinical pregnancy rates in female smokers. Despite an increased awareness of the adverse effects of cigarette smoke exposure on systemic health, many women remain unaware of the adverse effects of cigarette smoke on female fertility. This issue is compounded by our limited understanding of the molecular mechanisms behind cigarette smoke induced infertility. In this study we used a direct nasal exposure mouse model of cigarette smoke-induced chronic obstructive pulmonary disease to characterise mechanisms of cigarette-smoke induced ovotoxicity. Cigarette smoke exposure caused increased levels of primordial follicle depletion, antral follicle oocyte apoptosis and oxidative stress in exposed ovaries, resulting in fewer follicles available for ovulation. Evidence of oxidative stress also persisted in ovulated oocytes which escaped destruction, with increased levels of mitochondrial ROS and lipid peroxidation resulting in reduced fertilisation potential. Microarray analysis of ovarian tissue correlated these insults with a complex mechanism of ovotoxicity involving genes associated with detoxification, inflammation, follicular activation, immune cell mediated apoptosis and membrane organisation. In particular, the phase I detoxifying enzyme cyp2e1 was found to be significantly up-regulated in developing oocytes; an enzyme known to cause molecular bioactivation resulting in oxidative stress. Our results provide a preliminary model of cigarette smoke induced sub-fertility through cyp2e1 bioactivation and oxidative stress, resulting in developing follicle depletion and oocyte dysfunction.

Photodynamic therapy for hair removal

Background: Unwanted hair is one of the most common medical problems affecting women of reproductive age inducing a lot of psychological stress and threatening their femininity and self-esteem. Old methods of removing unwanted hair include shaving, waxing, chemical depilation, and electrolysis, all of which have temporary results. However laser-assisted hair removal is the most efficient method of long-term hair removal currently available. It is desirable to develop a reduced cost photodynamic therapy (PDT) system whose properties should include high efficiency and low side-effects. Method: Mice skin tissues were used in this study and divided into six groups such as controls, free methylene blue (MB) incubation, liposome methylene blue (MB) incubation, laser without methylene blue (MB), free methylene blue (MB) for 3 and 4 h and laser, liposome methylene blue (MB) for 3 h and laser. Methylene blue (MB) was applied to wax epilated areas. The areas were irradiated with CW He-Ne laser system that emits orange-red light with wavelength 632.8 nm and 10 mW at energy density of 5 J/cm2 for 10 min. The UV-visible absorption spectrum was collected by Cary spectrophotometer. Results: Methylene blue (MB) is selectively absorbed by actively growing hair follicles due to its cationic property. Methylene blue (MB) untreated sections showed that hair follicle and sebaceous gland are intact and there is no change due to the laser exposure. Free methylene blue (MB) sections incubated for 3 h showed that He:Ne laser induced destruction in hair follicles, leaving an intact epidermis. Treated section with free methylene blue (MB) for 4 h showed degeneration and necrosis in hair follicle, leaving an intact epidermis. Liposomal methylene blue (MB) sections incubated for 3 h showed He:Ne laser induced destruction in hair follicles with intradermal leucocytic infiltration. Conclusion: Low power CW He:Ne laser and methylene blue (MB) offered a successful PDT system in selectively damaging hair follicles, leaving an intact epidermis. The current PDT system provides better outcome than hair destruction through laser heat transfer procedures and laser-mediated hair removal, due to complete destruction of hair follicles.

Amiodarone-induced thyrotoxicosis

Aims Amiodarone is an iodine-rich drug widely used for the management of various cardiac arrhythmias and severe congestive heart failure. Thyroid dysfunction is an important side-effect of amiodarone therapy. Amiodarone-induced thyrotoxicosis is a complex diagnostic and therapeutic challenge. It can occur in apparently normal thyroid glands and in glands with pre-existing abnormalities. The aim of this study was to investigate the histo-pathological and immunohistochemical features of a total thyroid-ectomy specimen in a 70-year-old man with amiodarone-induced thyrotoxicosis. Methods Paraffin embedded thyroid tissues were stained with haematoxylin and eosin and special immunohistochemical and histochemical stains. Results Histological examination revealed destruction of the thyroid follicles with areas of degeneration and involution of the follicles. Together with chronic inflammatory changes and fibrosis, foamy cells containing fine-brown pigments filled approximately 50% of the follicles. Discussion The histological features of our case were in keeping with amiodarone-induced thyrotoxicosis.

The Spectrum of Histopathological Lesions in Scarring Alopecia: A Prospective Study

The aim of the article was to study the histopathological features of various lesions of Scarring Alopecia (SA) and to classify Primary SA on the basis of the predominant type of inflammatory cell component. Scarring or cicatricial alopecias are those that are produced as a result of the malformation, damage or destruction of the pilosebaceous follicles, which are replaced by cicatricial tissue, in such a way that they cannot again produce hair. This prospective study included 32 biopsy - proven cases of SA, who had attended our hospital. Primary SA was classified according to the North American Hair Research Society. The informed consents of the subjects and the institutional ethical clearance was obtained for the study. The SPSS, version 14 software was used to analyse the data. Frequencies and percentages were used to describe the data. During the study period, 32 cases of scarring alopecia were diagnosed, of which 24 were primary SA and 8 were secondary SA. Among the primary SA, there were 23 cases of lymphocyte associated primary scarring alopecias, of which, 19 of lupus erythematosus, 3 of lichen planopilaris (LPP) and one case of non specific SA. 1 case of neutrophil associated primary scarring (folliculitis decalvans) was also noted and among the secondary SA, there were 4 cases of morphea and 1 case each of lupus vulgaris, congenital absence of skin, burn and sarcoidosis. To conclude, histopathology is a dependable tool for identifying the underlying cause in scarring alopecia, which is helpful for an early diagnosis and treatment.

Long-Term Removal of Unwanted Hair Using Light

Laser (or light) hair removal, also referred to as photoepilation, is the most commonly used laser or light-based cosmetic medical procedure. The extended theory of selective photothermolysis is the basic principle for destruction of hair follicles using light. In this type of laser application the chromophore is follicular melanin. Several types of lasers and light sources have been effective for hair reduction, including the ruby, alexandrite, diode, and neodymium:yttrium-aluminum-garnet lasers and broadband, intense pulsed light sources. This article provides a broad overview of how hair can be removed using light, with an emphasis on practical considerations.

Interferon-γ inhibits integrin-mediated extracellular signal-regulated kinase activation stimulated by fibronectin binding in thyroid cells.

Hashimoto's thyroiditis (HT) is an autoimmune disorder characterized by the presence of specific antibodies and by a lymphocytic infiltration of the thyroid secreting inflammatory cytokines. Macrophages, lymphocytes, and cytokines play a pivotal role in both development and progression of Th1-mediated autoimmune diseases, and a direct role in the destruction of thyroid follicles and follicular cell function in autoimmune thyroiditis. Integrins are integral membrane receptors involved in cell-extra-cellular matrix (ECM) interaction with both structural and signaling functions. The integrin- ECM interaction is necessary for the correct function and survival of thyroid follicular cells. The purpose of this study was to determine the effect of cytokine stimulation on integrin expression and signaling in the thyroid cell. Primary cultures from normal thyroids were treated with interferon-γ (IFN-γ), INF-α, tumor necrosis factor-α, interleukin 1a or these cytokines all together. Integrin expression, cell adhesion to fibronectin (FN) and FN-stimulated extracellular signal-regulated kinase (ERK) phosphorylation were determined after cytokine treatment. IFN-γ and IFN-α were the most effective, reducing the expression of the integrin αvβ3 and slightly increasing the α3β1. Cell treatment with IFN-γ strongly impaired cell adhesion to FN. At the same time, the treatment with IFN-γ dramatically inhibited the stimulation of ERK phosphorylation induced by cell adhesion to FN. In conclusion, IFN-γ inhibits the expression of the integrin αvβ3, reducing the cell adhesion to FN and the following intracellular signaling in thyroid cells in culture. These results suggest that integrins may be a target of the infiltrating lymphocytes and have a role in the pathogenesis of autoimmune thyroiditis.

Long-term effects of thyroid fine-needle biopsy on the thyroid-related biochemical parameters

Thyroid fine-needle biopsy (FNB) is a simple, reliable, inexpensive and generally safe diagnostic procedure in the management of thyroid nodules. FNB may trigger biochemical alterations through destruction of thyroid follicles. We aimed to investigate long-term post-FNB alterations in serum thyroid-related parameters. One hundred and ten consecutive patients with thyroid nodular disease were subjected to FNB. Thyroid stimulating hormone (TSH), free thyroxine (FT4) and free triiodothyronine (FT3), thyroglobulin (Tg), thyroglobulin autoantibodies (anti-Tg), thyroid-peroxidase autoantibodies (anti-TPO) were measured in all subjects at baseline, 10 days, 2 and 6 months. Subsequently, patients were divided into subgroups according to the technique of FNB, the presence of disease characteristics as thyroid autoimmunity (Hashimoto's thyroiditis), goitre, singularity-maximum diameter-blood pattern of the nodule(-s), the number of passes and the administration of L-thyroxine (LT4). A significant increase in Tg, anti-Tg and FT3 levels was observed. These alterations were more prominent within patients with dominant nodule's maximum diameter ≥ 2 cm or without Hashimoto's thyroiditis. Tg and anti-Tg levels were significantly increased only in patients not being on LT4. On the other hand, FNB technique did not affect any of the measured parameters. Our data suggest that FNB results in statistically significant but clinically insignificant increases in Tg, anti-Tg and FT3 levels, implying a thyroid trauma of some level, more likely to happen in patients with larger nodules. The FNB technique used has no effect on the thyroid-related biochemical parameters.

Follicular B cells, not Marginal zone B cells, preferentially contribute to development of spontaneous autoimmune thyroiditis (SAT) and are targets of anti-CD20 therapy (56.12)

Abstract SAT in NOD.H-2h4 mice is a chronic organ-specific autoimmune disease characterized by mononuclear cell infiltration of the thyroid and destruction of thyroid follicles by infiltrating inflammatory cells. Like other autoimmune diseases, B cells play a critical role in pathogenesis of SAT as important antigen presenting cells (APCs) for activation of CD4+ T cells. However it is not yet known which subsets of B cells are important for SAT development. In this study, we found that expression of the co-stimulatory molecules CD80 and CD86 was significantly increased on splenic CD23hiCD21low follicular (FO) B cells, but not marginal zone (MZ) B cells, after SAT development, but absolute numbers and percentages of splenic FO B cells were not altered. In draining lymph nodes, CD40 expression on FO B cells was also increased in mice with SAT. All thyroid infiltrating B cells in mice with SAT were FO B cells. Depletion of FO B cells by anti-CD20 antibody ameliorated established SAT, and anti-thyroglobulin antibody levels were reduced. Moreover, administering anti-CD20 to mice with established SAT results in depletion of B cells in the thyroid and depletion of thyroid B cells is longer lasting than depletion of splenic B cells. On the basis of these findings, we suggest the FO subset of B cells preferentially contributes to SAT development and persistence, and targeting FO B cells after disease develops provides an effective means to treat autoimmune diseases.

Epidermal Cadm1 Expression Promotes Autoimmune Alopecia via Enhanced T Cell Adhesion and Cytotoxicity

Autoimmune alopecia is characterized by an extensive epidermal T cell infiltrate that mediates hair follicle destruction. We have investigated the role of cell adhesion molecule 1 (Cadm1; Necl2) in this disease. Cadm1 is expressed by epidermal cells and mediates heterotypic adhesion to lymphocytes expressing class 1-restricted T cell-associated molecule (CRTAM). Using a murine autoimmune alopecia model, we observed an increase in early-activated cytotoxic (CD8-restricted, CRTAM-expressing) T cells, which preferentially associated with hair follicle keratinocytes expressing Cadm1. Coculture with Cadm1-transduced MHC-matched APCs stimulated alopecic lymph node cells to release IL-2 and IFN-γ. Overexpression of Cadm1 in cultured human keratinocytes did not promote cytokine secretion, but led to increased adhesion of alopecic cytotoxic T cells and enhanced T cell cytotoxicity in an MHC-independent manner. Epidermal overexpression of Cadm1 in transgenic mice led to increased autoimmune alopecia susceptibility relative to nontransgenic littermate controls. Our findings reveal that Cadm1 expression in the hair follicle plays a role in autoimmune alopecia.

A Mouse Model of Clonal CD8+ T Lymphocyte-Mediated Alopecia Areata Progressing to Alopecia Universalis

Alopecia areata is among the most prevalent autoimmune diseases, yet compared with other autoimmune conditions, it is not well studied. This in part results from limitations in the C3H/HeJ mouse and DEBR rat model systems most commonly used to study the disease, which display a low frequency and late onset. We describe a novel high-incidence model for spontaneous alopecia areata. The 1MOG244 T cell expresses dual TCRA chains, one of which, when combined with the single TCRB present, promotes the development of CD8(+) T cells with specificity for hair follicles. Retroviral transgenic mice expressing this TCR develop spontaneous alopecia areata at nearly 100% incidence. Disease initially follows a reticular pattern, with regionally cyclic episodes of hair loss and regrowth, and ultimately progresses to alopecia universalis. Alopecia development is associated with CD8(+) T cell activation, migration into the intrafollicular region, and hair follicle destruction. The disease may be adoptively transferred with T lymphocytes and is class I and not class II MHC-dependent. Pathologic T cells primarily express IFNG and IL-17 early in disease, with dramatic increases in cytokine production and recruitment of IL-4 and IL-10 production with disease progression. Inhibition of individual cytokines did not significantly alter disease incidence, potentially indicating redundancy in cytokine responses. These results therefore characterize a new high-incidence model for alopecia areata in C57BL/6J mice, the first to our knowledge to apply a monoclonal TCR, and indicate that class I MHC-restricted CD8(+) T lymphocytes can independently mediate the pathologic response.

Understanding the Villain: DMBA-Induced Preantral Ovotoxicity Involves Selective Follicular Destruction and Primordial Follicle Activation through PI3K/Akt and mTOR Signaling

7,12-Dimethylbenz-[a]anthracene (DMBA) is an environmental carcinogen which has a potent ovotoxic affect on rat and mouse ovaries, causing complete follicular depletion resulting in premature ovarian failure. Although the overall effects of DMBA on ovarian folliculogenesis are well known, little is known about the exact molecular mechanisms behind its ovotoxicity. In this study, we characterized the mechanisms behind DMBA-induced ovotoxicity in immature follicles. Microarray analysis of neonatal mouse ovaries exposed to DMBA in vitro revealed a multilayered mechanism of DMBA-induced neonatal ovotoxicity involving a distinct cohort of genes and ovarian signaling pathways primarily associated with follicular atresia, tumorigenesis, and follicular growth. Histomorphological and immunohistological analysis supported the microarray data, showing evidence of primordial follicle activation and preantral follicle atresia both in vitro and in vivo. Further immunohistological analysis identified increased Akt1 phosphorylation, mTOR activation, and decreased FOXO3a expression in DMBA-treated primordial oocytes. Our results reveal a novel mechanism of DMBA-induced preantral ovotoxicity involving selective immature follicle destruction and primordial follicle activation involving downstream members of the PI3K/Akt and mTOR signaling pathways.

DNA Repair Gene Expression Up-Regulation in Response to Phosphoramide Mustard Exposure in Cultured Rat Ovaries.

Cyclophosphamide (CPA) is used as an anti-neoplastic and immunosuppressant therapy in females. Phosphoramide Mustard (PM) is the ovotoxic metabolite of CPA, and destroys rapidly dividing cells by binding covalently to DNA, inducing DNA-DNA and DNA-protein crosslinks, and DNA double-strand breaks (DSB). Previous studies have shown that PM induces destruction of primordial and small primary follicles in a dose-dependent manner in cultured rat ovaries. Interestingly, PM induced follicle destruction is preceded by the appearance of DSBs in oocytes. This study was designed to explore the expression of genes involved in DNA repair at time points prior to the appearance of PM-induced DSBs. Fisher 344 postnatal day 4 rat ovaries were cultured in medium containing vehicle control (DMSO) ± PM (30 µM) for 2 or 4 days. Three biological replicates were used to perform a real-time RT-PCR array designed to identify genes involved in DNA repair. The data were analyzed using mixed models, correcting for multiple testing by lme4 and q value packages in R software. Relative to control treated ovaries, 18 and 3 repair genes were up-regulated (q < 0.1) on days 2 and 4, respectively, in response to PM exposure. A single gene, DNA polymerase delta 3 (Pold3) was increased at both time points. These results support that the ovary up-regulates genes involved in DNA repair in response to a DNA damaging xenobiotic exposure. It is likely that repeated ovotoxic chemical exposure eventually overwhelms this protective response, resulting in loss of oocytes containing imperfect DNA. (Supported by ES016818 to AFK). (poster)

Perda pilosa por líquen plano pilar após transplante capilar: relato de dois casos e revisão da literatura

Androgenetic alopecia is often treated by follicular unit transplantation, a technique that involves minimal risk of hair loss because of the more resistant nature of the donor area. Lichen planopilaris is a cicatricial alopecia that causes permanent destruction of hair follicles. We report two cases of post-transplantation lesions compatible with lichen planopilaris in both recipient and donor areas. The quality of the hair follicles in the donor area was apparently compromised by lichen planopilaris, the probable cause of hair loss. Similar reports are rare. When lichen planopilaris is suspected, a biopsy of the scalp must be performed to avoid transplantation during disease activity.

Delineating Immune-Mediated Mechanisms Underlying Hair Follicle Destruction in the Mouse Mutant Defolliculated

Defolliculated (Gsdma3(Dfl)/+) mice have a hair loss phenotype that involves an aberrant hair cycle, altered sebaceous gland differentiation with reduced sebum production, chronic inflammation, and ultimately the loss of the hair follicle. Hair loss in these mice is similar to that seen in primary cicatricial, or scarring alopecias in which immune targeting of hair follicle stem cells has been proposed as a key factor resulting in permanent hair follicle destruction. In this study we examine the mechanism of hair loss in GsdmA3(Dfl)/+ mice. Aberrant expression patterns of stem cell markers during the hair cycle, in addition to aberrant behavior of the melanocytes leading to ectopic pigmentation of the hair follicle and epidermis, indicated the stem cell niche was not maintained. An autoimmune mechanism was excluded by crossing the mice with rag1-/- mice. However, large numbers of macrophages and increased expression of ICAM-1 were still present and may be involved either directly or indirectly in the hair loss. Reverse transcriptase-PCR (RT-PCR) and immunohistochemistry of sebaceous gland differentiation markers revealed reduced peroxisome proliferator-activated receptor-γ (PPARγ), a potential cause of reduced sebum production, as well as the potential involvement of the innate immune system in the hair loss. As reduced PPARγ expression has recently been implicated as a cause for lichen planopilaris, these mice may be useful for testing therapies.

AA amyloidosis associated with macroglobulinemia

AA amyloidosis has been reported to be associated with chronic inflammatory diseases including rheumatoid arthritis, juvenile rheumatoid arthritis and chronic infection [1]. However, there have been only a few reports on patients with AA amyloidosis associated with hematological tumors [2–4]. In this report, we present a case of AA amyloidosis associated with macroglobulinemia in which combination chemotherapy was effective for amyloidosis and markedly improved the condition of the patient. A 44-year-old man was admitted to our hospital because of a 3-month history of fever and lymphadenopathy. Hematological examination revealed a normal white blood cell (WBC) count (4.3 9 10/L) with 17% lymphocytes and a normal platelet count (366 9 10/L) but a slight decrease in hemoglobin concentration (12.6 g/dl). Laboratory examinations disclosed high levels of IgM (880 mg/dl), b2-microglobulin (2.2 mg/l) and soluble interleukin-2 receptor (3,460 U/ml). Serum and urine immunoelectrophoresis demonstrated IgM-j type of M-protein and Bence Jones protein-j, respectively. CT scanning revealed hepatosplenomegaly as well as swelling of para-aorta lymph nodes. Histological examination of an excised cervical lymph node showed destruction of lymphatic follicles by plasma cell infiltration, which is not contradictory to lymphoplasmacytoid lymphoma. In addition, flow cytometric analysis of the lymph node sample showed 52.7% of CD20? cells, 41.3% of IgM? cells, 48% of j? cells and 9.2% of k? cells, indicating monoclonal proliferation of B cells with IgM-j. Furthermore, a bone marrow aspiration sample showed hypercellularity with an increased number of mature lymphocytes (66%). Phenotypes of these lymphocytes are similar to those of the lymph node cells. Based on the results, we diagnosed this case as macroglobulinemia. Importantly, eosinophilic substances, which were positive for Congo-Red staining with green birefringence under a polarizing microscope, were also observed in the lymph node (Fig. 1a). These substances were stained with anti-AA antibody by immunohistological staining, whereas no staining was present with both anti-j and anti-k antibodies. Based on these findings, the substances were defined as AA amyloid (Fig. 1b). AA amyloid was also detected in biopsy samples of bone marrow, stomach and rectum. In addition to the deposition of AA amyloid in various organs, the level of serum amyloid A protein (SAA) was extremely high (99.9 mg/ml). The reason for the elevation of SAA was not clear; however, it is possible that macroglobulinemia itself induced inflammation, resulting in an increase in the level of SAA in this case. Based on these findings, the patient was diagnosed as having systemic AA amyloidosis. Since there was no evidence of tuberculosis or collagen diseases including rheumatoid arthritis, the AA amyloidosis was thought to be associated with macroglobulinemia. The patient was treated with 30 mg prednisolone as an initial treatment. After 1 month of this therapy, both fever and lymph node swelling were improved. However, lymphadenopathy recurred with dose reduction of prednisolone. In addition, continuous increases in hepatomegaly and levels of alkaline phosphatase (ALP) and c-glutamyl transpeptidase (c-GTP), probably due to infiltration of R. Tatara T. Nagai (&) H. Kobayashi K. Hatano T. Suzuki K. Ozawa Division of Hematology, Jichi Medical University, 3311-1 Yakushiji, Shimotsuke, Tochigi 329-0498, Japan e-mail: t-nagai@jichi.ac.jp

Combination of gonadotropin-releasing hormone (GnRH) agonists with GnRH antagonists before chemotherapy reduce but does not completely prevent a follicle-stimulating hormone flare-up

Increasing evidence supports GnRH agonists to be an effective treatment to preserve ovarian function during chemotherapy, but the initial flare-up of FSH during the first week after GnRH agonist application still limits its use. The combination of GnRH agonists with GnRH antagonists might solve this problem to some extent as the addition of GnRH antagonists at least significantly reduces the FSH flare-up.

Primary cicatricial alopecias

Primary cicatricial alopecias are uncommon folliculocenteric inflammatory disorders that represent a ‘trichologic emergency’. Unless correctly diagnosed and abruptly treated, these diseases can lead to permanent destruction of hair follicles and irreversible hair loss. Based on the predominant inflammatory cell type present, they are classified into lymphocytic, neutrophilic and mixed. The etiology and pathogenesis of many of these disorders are incompletely understood. A systematic review of classification, pathogenesis, clinical features, histopathologic findings and therapeutic options will be highlighted in this article.

Prospective controlled clinical and histopathologic study of hidradenitis suppurativa treated with the long-pulsed neodymium:yttrium-aluminium-garnet laser

Hidradenitis suppurativa (HS) is a chronic inflammatory disease involving the intertriginous areas. We sought to conduct clinical and histopathologic evaluation of the efficacy of long-pulsed neodymium:yttrium-aluminium-garnet laser treatment for HS. We conducted a prospective, randomized, right-left within-patient controlled trial for HS (n = 22). Four monthly laser sessions were performed. Disease activity was measured at baseline, and treatment response was assessed before each laser session and monthly for 2 months after the completion of laser treatment, using a modified scoring system based on Sartorius score. Histologic examination was performed at baseline, immediately after laser treatment, and at 1 and 4 weeks after treatment. A patient questionnaire was circulated on the last visit to assess patients' level of satisfaction. There was progressive improvement in disease activity, most significantly during the 4 months of treatment, which was maintained during the 2-month posttreatment follow-up period. Averaged over all anatomic sites, the percent improvement was 72.7% on the laser treated side, and 22.9% on the control side (P < .05). Histologic examination showed an initial acute neutrophilic infiltrate. Granulomatous inflammation was present on follow-up biopsy specimens 4 weeks later. An inflammatory infiltrate surrounded the hair shaft remnants, denoting destruction of hair follicles. Small sample size was a limitation. Long-pulsed neodymium:yttrium-aluminium-garnet laser, together with topical benzoyl peroxide and clindamycin, is significantly more effective than topical benzoyl peroxide and clindamycin alone for the treatment of HS. Preliminary review of histopathology suggests the mechanism of action is destruction of the hair follicle. The overall success of the treatment in both clearing pre-existing lesions and preventing new eruptions, coupled with high patient satisfaction, makes the neodymium:yttrium-aluminium-garnet laser a promising treatment advance for this highly disabling condition.

Thrombospondin-1 Inhibits Angiogenesis and Promotes Follicular Atresia in a Novel in Vitro Angiogenesis Assay

Thrombospondin-1 (TSP-1) is a putative antiangiogenic factor, but its role in regulating physiological angiogenesis is unclear. We have developed a novel in vitro angiogenesis assay to study the effect of TSP-1 on follicular angiogenesis and development. Intact preantral/early antral follicles dissected from 21-d-old rat ovaries were cultured for 6 d in the presence or absence of TSP-1. At the end of the culture period, angiogenic sprouting from the follicles was quantified using image analysis. Follicles were fixed and sectioned, and follicular apoptosis was assessed by immunohistochemistry for activated caspase-3 in granulosa cells. The results showed that TSP-1 inhibited follicular angiogenesis (P < 0.01) and promoted follicular apoptosis (P < 0.001) in a dose-dependent manner. To determine whether the proapoptotic activity of TSP-1 is mediated by direct effects on granulosa cells, isolated granulosa cells were cultured with TSP-1 (0, 10, 100, and 1000 ng/ml) for 48 h. Apoptosis was quantified using a luminescent caspase-3/7 assay. TSP-1 promoted apoptosis of granulosa cells in a dose-dependent manner (P < 0.05), suggesting that TSP-1 can act independently of the angiogenesis pathway to promote follicular apoptosis. These results show that TSP-1 can both inhibit follicular angiogenesis and directly induce apoptosis of granulosa cells. As such, it may have potential as a therapeutic for abnormal ovarian angiogenesis and could facilitate the destruction of abnormal follicles observed in polycystic ovary syndrome.