Evaluation of Gene Expression Patterns in Micrografts Demonstrate Induction of Catagen-Like Processes During Storage.

BACKGROUND: Alterations of gene expression patterns may contribute to the commonly observed transient reduction of hair shaft elongation in hair restoration surgery. OBJECTIVE: To elucidate the molecular causes, we evaluated changes in gene expression patterns in hair follicle micrografts during storage. MATERIALS AND METHODS: Micrografts with different amounts of adjacent connective tissue (regular, skinny, and chubby) were stored for different periods, and the expression of key genes was determined: dermal papilla (DP): FGF7, alkaline phosphatase (ALP), versican; outer root sheath: Krt15; inner root sheath: Krt 25; cuticula: Krt85; Henle layer: filaggrin; genes related to apoptosis and growth/differentiation: Caspase 3, Ovol1, and Foxo1. RESULTS: A significant decrease in FGF7 (4 × 10 fold) was observed after 4 hours, with further decrease after 48 hours. A significant decrease of versican (0.35 fold) and ALP (0.004 fold) was observed after 24 hours of storage. No differences relating to adjacent connective tissue were observed. No changes of different keratins genes or genes related to growth/differentiation and apoptosis were observed. CONCLUSION: These data clearly demonstrate a reduction in the specific function of cells in the DP, which seem to be the causative for the induction of hair follicle cycling during micrograft preparation and storage.

Highly efficient and compatible shampoo for use after hair transplant.

BACKGROUND: Sensitive or hyperreactive skin is a common condition defined by prickling, burning, pain, and pruritus. Although this skin problem was initially described on the face, the scalp is often affected. A sensitive scalp can react with irritation to harsh surfactants or other additives which are often present in shampoos. For this reason, we developed a new rinse-off hypertolerant shampoo specifically designed for the hypersensitive and problematic scalp. METHODS: The shampoo formulation is based on an extremely mild surfactant system and contains bisabolol, an anti-irritant and anti-inflammatory ingredient of chamomile. The shampoo is free of additives such as perfumes, silicones, colorants, parabens, paraffins, and betaine. Since skin can remain in a hyperreactive state after wounding, the status after hair transplantation was chosen as a model system to test the shampoo. Scalp condition and compatibility of each volunteer were analyzed by a plastic surgeon directly after hair transplant and after stitch removal. The plastic surgeons also rated whether they would recommend the further use of the test shampoo. Additionally, volunteers completed a self-assessment questionnaire. RESULTS: Following hair transplantation, regular use of the shampoo resulted in a significant reduction in the extent of scabbing and erythema. This was confirmed by dermatological scalp examinations performed by the plastic surgeon as well as in volunteers' self-assessments. The plastic surgeon highly recommended the further use of the test shampoo after hair transplant to all study participants. CONCLUSION: Application of the test shampoo demonstrated excellent skin compatibility and product efficacy after hair transplant. The test shampoo significantly reduced the extent of scabs and erythema. Therefore, the shampoo is ideally suited for use after hair transplantation and for the treatment of sensitive scalp. The excellent skin compatibility is because of the mild surfactant system, the calming ingredient bisabolol, and the absence of potentially irritating ingredients.

Nucleocytosolic depletion of the energy metabolite acetyl-coenzyme a stimulates autophagy and prolongs lifespan.

Healthy aging depends on removal of damaged cellular material that is in part mediated by autophagy. The nutritional status of cells affects both aging and autophagy through as-yet-elusive metabolic circuitries. Here, we show that nucleocytosolic acetyl-coenzyme A (AcCoA) production is a metabolic repressor of autophagy during aging in yeast. Blocking the mitochondrial route to AcCoA by deletion of the CoA-transferase ACH1 caused cytosolic accumulation of the AcCoA precursor acetate. This led to hyperactivation of nucleocytosolic AcCoA-synthetase Acs2p, triggering histone acetylation, repression of autophagy genes, and an age-dependent defect in autophagic flux, culminating in a reduced lifespan. Inhibition of nutrient signaling failed to restore, while simultaneous knockdown of ACS2 reinstated, autophagy and survival of ach1 mutant. Brain-specific knockdown of Drosophila AcCoA synthetase was sufficient to enhance autophagic protein clearance and prolong lifespan. Since AcCoA integrates various nutrition pathways, our findings may explain diet-dependent lifespan and autophagy regulation.

Randomised controlled trial with medical leeches for osteoarthritis of the knee.

OBJECTIVES: To evaluate the possible efficacy of medical leeches (Hirudo medicinalis) in the treatment of patients with active osteoarthritis of the knee. DESIGN: Unblinded, randomised controlled trial with outpatients in a crossover design with single interventions of either leeches or transcutaneous electrical nerve stimulation (TENS) as comparator. MAIN OUTCOME MEASURES: Change in Lequesne's combined index for pain and function and change (L.I.) and overall assessment of complaints by visual analog scale (VAS). Cross-over at day 42, with further observation period of 21 days. RESULTS: 52 out of 72 screened patients were randomised (intent to treat) to initial treatment with either eight leeches (group 1: 27 patients) or TENS (group 2: 25 patients). Due to phase effects, confirmatory evaluation had to be restricted to the first period. Between days 0 and 21, we observed highly significant (p<0.001) improvements for means of Lequesne's index from 12.07 to 9.37 and for VAS from 5.89 to 4.16 cm for leeches, but no significant differences for TENS. Effect size as group difference was -2.50 for L.I. (95% confidence interval -3.88 to -1.11), resp. -1.86 cm for VAS (95% confidence interval -2.85 to -0.87 cm). 12 patients (5 group 1, 7 group 2) did not finish the trial, mostly due to non-compliance (6). No serious adverse effects were observed. CONCLUSIONS: Single leech therapy showed significant, relevant and sustaining effects, comparable to other trials with leeches. The method deserves further research, esp. into mechanisms of possible specific effects and optimization of dosing by number of leeches and possible repeats.

Reorganization of hair follicles in human skin organ culture induced by cultured human follicle-derived cells.

Studies of human hair follicle (HF) induction by follicle-derived cells have been limited due to a lack of suitable test systems. In this study, we established a skin organ culture system which supports HF formation by follicle-derived cells. Long-term skin organ cultures were set up from human retroauricular skin specimens and maintained in culture for up to 8 weeks. In vitro expanded human HF-derived cells from the dermal papilla (DP) and the outer root sheath (ORS) were injected together into the skin specimens and evaluated for their ability to induce reorganization of HFs. Macroscopic analysis of the cultured skin specimens demonstrated the growth of velus-like hair after 4 weeks in culture. Histologic evaluation of the cultured skin specimens after 8 weeks of culture revealed multiple miniaturized HFs with sebaceous glands. In addition, cell clusters of various differentiation stages could be demonstrated in serial sections of the cultured skin specimens. Labeling of HF-derived cells with the fluorescence dye CFDA-1 prior to injection suggested a de novo reorganization of HFs out of the injected cells. In conclusion, the study demonstrated HF formation by HF-derived cells in an in vitro skin organ culture model.

Induction of vascular endothelial growth factor messenger ribonucleic acid expression in stored micrografts by aminoguanidine.

BACKGROUND AND OBJECTIVES: Aminoguanidine (AMG) has been found to inhibit apoptotic cell death in hair follicle micrografts and improves the viability of isolated micrografts during the storage period in hair restoration surgery. In this study, we investigated the effect of AMG on messenger ribonucleic acid (mRNA) synthesis of growth factors in stored micrografts and primary cultures of follicle-derived cell populations. METHOD: Hair follicles were obtained from 10 different patients undergoing routine micrograft transplant and were stored for 5 hours at room temperature in phosphate-buffered saline containing different concentrations of AMG. After a culture period of 72 hours, quantitative changes of mRNA for basic fibroblast growth factor (bFGF), insulin-like growth factor 1 (IGF-1), and vascular endothelial growth factor (VEGF) were determined by real-time reverse transcriptase-polymerase chain reaction (RT-PCR). Primary cell cultures of dermal papilla and outer root sheath cells were stimulated for 72 hours with AMG followed by RT-PCR measurement of growth factor mRNA. RESULTS: A dose-dependent induction of VEGF mRNA could be demonstrated in stored micrografts after stimulation with AMG (unstimulated: 1.0 [0.7-2.2]; AMG 10 pg/mL: 5.6 [3.1-9.7], p < .05; AMG 50 pg/mL: 6.9 [5.7-10.0], p < .05; AMG 100 microg/mL: 17.1 [14.1-22.3], p < .001). Expression of bFGF mRNA and IGF-1 mRNA levels was not influenced by AMG stimulation. Stimulation of cultured dermal papilla and outer root sheath cells demonstrated 14-fold induction of VEGF mRNA by AMG in outer root sheath cells (unstimulated: 1.0 [0.8-1.4]; AMG 100 pg/mL: 14.0 [12.5-16.1], p < .01), and no changes in VEGF mRNA levels were detected in dermal papilla cells. CONCLUSIONS: Our study demonstrated induction of VEGF mRNA in stored micrografts by AMG. Although the clinical relevance in post-transplant hair growth and wound healing needs further evaluation, the possibility of actively influencing growth factor production in isolated micrografts during the storage period is the basis for the development of hair follicle growth-promoting storage solutions in the future.

Enhancement of in vitro hair shaft elongation in follicles stored in buffers that prevent follicle cell apoptosis.

BACKGROUND: Viability and survival of stored micrografts during hair follicle transplantation are important limitations of micrograft transplantation procedures. In this study, we investigated the effect of different storage solutions and inhibitors of apoptotic cell death (ACD) on hair follicle cell viability by measuring in vitro hair shaft elongation (HSE) for 5 days. METHODS: Micrografts from informed patients undergoing routine micrograft transplantation were stored for 5 hours at room temperature in phosphate-buffered salt solution (PBS) or HEPES-buffered Dulbecco's modified Eagle's medium (DMEM), containing different concentrations of the ACD-inhibitors aminoguanidine (AMG), hormones (insulin, hydrocortisone), 14,15-epoxy-eicosatrienoic acid (14,15-EET), or combinations of these. RESULTS: In vitro, HSE was significantly increased in micrografts stored in DMEM compared with PBS (2.3%+/-0.6% vs. 28.4%+/-3.9%, P<0.0001). DMEM supplemented with AMG (10 microg/mL) or 14,15-EET (1 ng/mL) further increased in vitro HSE (33.9%+/-7.1%, p=0.01, and 32.8%+/-6.1%, P=0.02, respectively). Evaluation of ACD in stored micrografts, performed by determination of cytoplasmic histone-associated DNA fragments, confirmed the results found by HSE. ACD was detectable after a 36-hour culture in serum-containing medium and was higher in micrografts stored in PBS compared with micrografts stored in DMEM (A405nm/A492nm: 1.63+/-0.21 vs. 1.42+/-0.07, respectively; P<0.01). The addition of AMG further decreased serum-induced ACD in the micrografts (DMEM 1.42+/-0.07 vs. DMEM/AMG 0.90+/-0.11, P<0.0001). CONCLUSION: Our study demonstrated an important role of ACD in micrograft transplantation surgery. Preconditioning of micrografts with storage buffers containing inhibitors of ACD could prevent serum-induced ACD after transplantation and might increase the viability of micrografts and the clinical outcome in micrograft transplantation.