毛囊生物学及其在皮肤组织工程中的应用

本文讨论了毛囊的胚胎发生和周期性生长特性,重点阐述了毛乳头细胞在毛囊生长过程中的重要作用,毛囊干细胞的特性及定位,以及毛囊生长发育的几种不同机制。通过回顾国外在毛囊生物学研究方面所取得的不同成就及其研究技术手段,探索构建人工复合皮上毛囊的方法和途径。     

STR genotyping of exogenous hair shaft DNA

Most hairs found at crime scenes yield low quality and/or low quantities of nuclear DNA. This DNA is further depleted when stringent hair cleaning procedures are applied in the laboratory, suggesting that detectable DNA exists exogenously. The phenomenon of exogenous hair DNA is the subject of this study. DNA was extracted from washed and unwashed hairs and the resulting Profiler? Plus STR genotypes were compared with those of reference (buccal) swabs from the hair donors. The DNA extraction procedure involved no prior cleaning of the hair sample and no dissolution of the hair during digestion, in contrast to standard procedures. The STR genotyping success was measured by recording the two dominant alleles at each locus and comparing them with the reference DNA profile. The effect of hair cleanliness was examined by leaving donors’ hair unwashed for periods of 1, 3 and 7 days before sampling. It was found that the genotyping success for unwashed hair was significantly higher than that for freshly washed hair, with the majority of clean hair samples producing little or no DNA. Genotyping success was also lower for donors with cosmetically treated hair compared with those having untreated hair. Although the quality of STR profiles (i.e. allele dropout, differential amplification) from hair shafts or telogen hair clubs is reduced compared with those from other biological sources, the genotypes obtained in this study may be usable and are certainly discriminating if alternative interpretational methods are applied.     

Zum Suchtmittelnachweis in Haaren IV. Einfluß der Pigmentierung auf den Ofloxacingehalt in Haaren bei Meerschweinchen

Tortoise shell guinea pigs (n = 6) were administered ofloxacin (1 mg/mL) in their drinking water for 3 weeks. Black, reddish-brown and white hair was collected separately from each animal before and after treatment. The hair samples were analyzed by HPLC. All hair fibers showed positive results for the drug substance administered. Even low drug intake resulted in distinct differences of the drug content in hair fibers of different color, whereas in cases of high drug intake an increasing influence of hair pigmentation on the analytical results was observed. The highest drug content was always found in black hair samples, non-pigmented hair showed the lowest drug concentrations and the drug content in reddish-brown fibers was less than in black hair samples from the same animal. From the results it was concluded, that eumelanins rather than pheomelanins are the decisive factor for ofloxacin-melanin binding in hair and the amount of drug intake is thought to determine the relevance of hair pigmentation on the analytical results.     

Methadone concentrations in human hair of the head,axillary and pubic hair

Summary The concentrations of methadone and its metabolites in the hair of the head, axillary and pubic hair obtained from patients receiving a daily maintenance doses, were determined. Comparison of the concentrations provides the highest values in the axillary hair, followed by pubic hair and the hair of the head.     

Dose–hair concentration relationship and pigmentation effects in patients on low-dose clozapine

Several hair components have been suggested as possible molecular sites for drug binding and interaction. Of these, keratin and melanin have been investigated in some detail in order to assess the mechanisms by which the binding occurs. Substances that are positively charged at physiological pH may interact by electrostatic forces between their cationic groups and the anionic carboxylic groups on the surface of the melanin polymer. Studies in human subjects with grey hair have shown that various drugs are detectable in both the coloured (melanin rich) and white (melanin free) hair shafts of these individuals. Again this supports the proposition that keratin and hair proteins play an important role in the binding of drugs in hair. However, drugs are often found in significantly higher concentrations in pigmented hair strands than in senile white hair strands. Another interesting question is if the concentration measured in hair reflects the dose taken. Previous reports have both verified and rejected this hypothesis, but most agree that many factors have impact on the incorporation rate, melanin being one. In this study we obtained blood and hair samples from 12 grey haired patients treated with low-dose clozapine as an adjunct medication in their treatment against Parkinson disease. Each patient’s hair was divided into a pigmented and a non-pigmented portion and those were analyzed separately. Clozapine and desmethylclozapine were analyzed with LC-MS-MS after extraction of the analytes from hair and plasma. Paired results from the analysis of pigmented and white hair confirmed the preference for binding to pigmented hair for both clozapine and its metabolite. A majority of the incorporated clozapine was found in the pigmented hair but, as drugs could be detected in white hair, binding to hair protein or association with other hair matrix account for a significant part of drug accumulation in hair. High correlations between dose and the measured concentration of analyte were found for both clozapine (r = 0.91) and desmethylclozapine (r = 0.88). The data were presented at the Hair Testing Meeting, Vadstena, Sweden, May 2006. We hereby state that we do not have a significant financial interest or other relationship with any product manufacturer or provider of services discussed in this article.     

In situ labeling of DNA reveals interindividual variation in nuclear DNA breakdown in hair and may be useful to predict success of forensic genotyping of hair

Hair fibers are formed by keratinocytes of the hair follicle in a process that involves the breakdown of the nucleus including DNA. Accordingly, DNA can be isolated with high yield from the hair bulb which contains living keratinocytes, whereas it is difficult to prepare from the distal portions of hair fibers and from shed hair. Nevertheless, forensic investigations are successful in a fraction of shed hair samples found at crime scenes. Here, we report that interindividual differences in the completeness of DNA removal from hair corneocytes are major determinants of DNA content and success rates of forensic investigations of hair. Distal hair samples were permeabilized with ammonia and incubated with the DNA-specific dye Hoechst 33258 to label DNA in situ. Residual nuclear DNA was visualized under the fluorescence microscope. Hair from some donors did not contain any stainable nuclei, whereas hair of other donors contained a variable number of DNA-positive nuclear remnants. The number of DNA-containing nuclear remnants per millimeter of hair correlated with the amount of DNA that could be extracted and amplified by quantitative PCR. When individual hairs were investigated, only hairs in which DNA could be labeled in situ gave positive results in short tandem repeat typing. This study reveals that the completeness of DNA degradation during cornification of the hair is a polymorphic trait. Furthermore, our results suggest that in situ labeling of DNA in hair may be useful for predicting the probability of success of forensic analysis of nuclear DNA in shed hair.     

Influence of the cosmetic treatment of hair on drug testing

An important issue of concern for drug analysis in hair is the change in the drug concentration induced by the cosmetic treatment of hair. The products used for this treatment are strong bases and they are expected to cause hair damage. As a result drugs may be lost from the hair matrix or, under conditions of environmental contamination, be more easily incorporated into the hair matrix. We investigated the effects of cosmetic treatment in vivo by analysing hair samples selected from people who had treated their hair by bleaching or dyeing before sample collection. All of the subjects admitted a similar drug consumption during the time period for which the strands were analysed. Samples were viewed under a microscope to establish the degree of hair damage. Treated and untreated portions from each lock of hair were then selected, separated and analysed by standard detection procedures for cocaine, opiates, cannabinoids and nicotine. In all cases the drug content in hair that had undergone cosmetic treatment decreased in comparison to untreated hair. The majority of the mean differences were in the range of 40%-60% (cocaine, benzoylecgonine, codeine, 6-acetylmorphine and THC-COOH). For morphine the mean difference was higher than 60%, and two cases (THC and nicotine) differed by approx. 30%. These differences depended not only on the type of cosmetic treatment, as bleaching produced higher decreases than dyeing, but also on the degree of hair damage i.e. the more damaged the hair, the larger the differences in the concentration levels of drugs. Received: 15 January 1996 / Received in revised form: 23 January 1997     

Meeting overview

Abstract

Purpose: The purpose of this study was to determine whether the effects of gamma rays on the regeneration of hair follicles are carried over to later hair cycles.

Materials and methods: The whole bodies of C57BL/10JHir mice in the 1st telogen phase were irradiated with ⁶⁰Co γ-rays. Mice were examined for the effects on hair follicles, including their number, morphology and pigmentation in the 3rd anagen phase. Effects of γ-rays on hair follicle stem cells were investigated by the indirect immunolabeling of keratin 15 (K15).

Results: Decreased hair follicle density and induction of curved hair follicles were observed in the dermis of irradiated skin. In addition, white hair and hypopigmented hair bulbs were found. The number of K15-positive hair follicle stem cells in the hair bulge region of irradiated skin appeared to decrease slightly but not significantly.

Conclusions: These results suggest that the effects of γ-rays are carried over to a later hair cycle to affect the number, structure and pigmentation of hair follicles in the 3rd anagen phase when stem cells and committed progenitors for keratinocytes and melanocytes are irradiated in the 1st telogen phase.     

不同条件对人游离毛囊培养的影响

目的：研究不同培养基对游离毛囊培养的影响。方法：将美容手术后取下的头皮在无菌条件下分离成游离的毛囊培养于含血清或无血清的DMEM培训基、WilliamsE培养基中,观测毛囊的生长速度及形态学改变。结果：在含血清DMEM培养基中毛囊生长速度较慢,扭曲变形早;而无血清DMEM培养基毛囊的生长天数明显延长,毛囊的层次结构及形态可长时间保不变,但前3天长速度与有血清培养时无明显差异;使用WlliamesE培养基毛囊在前3天生长速度明显加快,结论：含血清的DMEM培养基不利于毛囊的生长,而WilliamsE培养基较适用于游离毛囊的培养。     

不同条件对人游离毛囊培养的影响

目的：研究不同培养基对游离毛囊培养的影响。方法：将美容手术后取下的头皮在无菌条件下分离成游离的毛 囊培养于含血清或无血清的DMEM培训基、Williams E培养基中,观测毛囊的生长速度及形态 学改变。结果：在含血清DMEM培养 基中毛囊生长速度较慢,扭曲变形早;而无血清DMEM培养基毛囊的生长天数明显延长,毛 囊的层次结构及形态可长时间保持不变,但前3天生长速度与有血清培养时无明显差异;使 用Wlliams E培养基毛囊在前3天生长速度明显加快。结论：含血清的DMEM培养基不利于毛囊的生长,而Williams E培养基较适用于游离毛囊的培养。     

800nm半导体激光真空脱毛系统的临床疗效观察

目的评价800 nm半导体激光真空脱毛系统的疗效及安全性。方法脱毛患者128例,应用800 nm半导体激光对不同部位,包括腋部、四肢、躯干部及毛表皮痣皮损处毛发进行脱毛治疗,并对效果及安全性进行评价。能量密度6～11 J/cm2,光斑大小22 mm×35 mm,脉宽30～400 ms,采用真空负压脱毛系统,根据患者肤色、毛发色泽和粗细调整治疗参数。结果 128例患者经3～6次连续治疗后均达到理想的脱毛效果,治疗后毛发明显减少,再生毛发细小、浅淡。腋部的总有效率为100.0%、双上肢为90.9%、双下肢为93.9%、躯干部皮损为83.3%、毛表皮痣为75.0%,随着治疗次数的增加总有效率不断提高,均未出现色素沉着或减退、瘢痕等不良反应。结论 800 nm半导体激光的真空脱毛系统是目前比较安全、高效、快速的脱毛方法。     

Two DFSA cases involving midazolam clarified by the micro-segmental hair analyses

Purpose

In this study, an analytical procedure to identify trace amounts of drug in hair based on micro-segmental hair analysis was presented. The method also can be used to estimate the time of drug ingestion at daily precision by cutting a single hair into sub-millimeter segments which correspond to daily hair growth.

Methods

A method was established for efficient extraction of midazolam, one of the most frequently detected compound in drug-facilitated sexual assault (DFSA) cases, from each 0.4-mm hair segment and validated by ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UPLC–MS/MS). Moreover, two DFSA cases were used to compare the micro-segmental hair analysis with the 1- cm segmental analysis method.

Results

The validation showed a lower limit of quantification of 0.5 pg/mm for midazolam, with intraday and interday accuracies (bias) from ???5.2 to 0.9%. The micro-segmental hair analysis method was applied to proximal 1-cm hair segment including hair bulbs in two DFSA cases. The micro-segmental hair analysis results in case 1 showed midazolam in the S15–S17 (5.6–6.8 mm from hair bulb) in a concentration range from 0.5 to 0.9 pg/mm, and the concentrations of midazolam in all hair micro-segments (0–1 cm from the scalp) in case 2 were from 0.5 to 2.0 pg/mm.

Conclusions

Comparison with the conventional method revealed that micro-segmental hair analysis may enhance the utility of hair drug testing and strengthen probative force in DFSA cases.

     

Analysis of a new fluoroquinolone derivative (Q-35) in human scalp hair as an index of drug exposure and as a time marker in hair

Summary Scalp hair samples were obtained every month for three months after administration from healthy male volunteers who participated in the phase I study of a new antimicrobial fluoroquinolone derivative (Q-35). Hairs were cut into 1 cm long pieces successively from the scalp end. Corresponding pieces of 5 hair strands were dissolved in 1M NaOH and assessed for Q-35 by HPLC. The drug was detectable in the hairs of all subjects taking either a single (400 mg,n = 6) or repeated oral doses of Q-35 (400 mg/day for 6.5 days, total 2600 mg,n = 6). The hair portions containing the drug were shown in most subjects to move outwards along the hair shafts month by month in proportion to the hair growth rate of about 1 cm/month. Q35 (600 mg/day) was also given to 6 healthy male volunteers for 6.5 days (total 3900 mg) and hair samples were obtained 1 and 3 months after administration. When Q-35 was analyzed along a single hair shaft, the drug was detectable only in 1–2 consecutive 1 cm long pieces, which were also shown to move outwards along the hair shaft with time. A detailed analysis revealed that the drug was contained only in 2–4 consecutive 2.5 mm long pieces of a single hair collected after 3 months, showing that there was no significant axial diffusion of the drug along the hair shaft with time. These findings indicate the utility of measuring this fluoroquinolone derivative in human scalp hair as an index of drug exposure and as a time marker for analyzing other drug(s) in hair.     

Effects of gamma rays on the regeneration of murine hair follicles in the natural hair cycle

Purpose: This review evaluates the effects of γ-rays on the regeneration of murine hair follicles in the natural hair cycle. A series of studies were performed to investigate this issue, in which the whole bodies of C57BL/10JHir mice in the 1st telogen phase of the hair cycle were irradiated with γ-rays.

Results: The dermis of the irradiated skin showed a decrease in hair follicle density and induction of curved hair follicles along with the presence of white hairs and hypopigmented hair bulbs in the 2nd and 3rd anagen phases. An increased frequency of hypopigmented hair bulbs was still observed in the later hair cycle at postnatal day 200. There was no significant difference in the number of stem cells in the hair bulge region between control and irradiated skin.

Conclusions: These results show that the effects of γ-rays on the pigmentation of murine hair follicles are persistently carried over to later hair cycles, although those on the number and structure of hair follicles appear to be hidden by the effects of aging. Our findings may be important for understanding the mechanisms of the actions of stem cells on hair regeneration in connection with age-related phenotypes.     

毛发移植进展

目的:综述毛发移植的研究进展。方法:采用文献回顾和综合分析方法。结果:毛发移植有许多新的术式、新设备出现,毛发移植的基础研究已达到分子和基因水平,然而毛囊干细胞的研究和毛囊体外培养的临床应用还很不成熟。结论:毛发组织工程学的重大突破将提供更好的毛发移植技术,对整形科技极具现实意义。     

Relationship between hair/eye color and primary vesicoureteral reflux in children

The relationship between hair/eye color and primary vesicoureteral reflux was analyzed in more than 900 children. Children with blonde hair/blue eyes did not show an increased prevalence of reflux nor did any other hair/eye combination. With the possible exception of rufous coloring, the color of the hair and eyes are poor predictors of the competence of the ureteral orifice.     

In vivo incorporation of fenfluramine and norfenfluramine into pigmented and nonpigmented hair of rats measured by HPLC-fluorescence detection

The incorporation profiles for fenfluramine (Fen) and its metabolite norfenfluramine (Norf) into black hair and white hair of Zucker rats and into white hair of Wistar rats after intraperitoneal (i.p.) administration of Fen or N-nitrosofenfluramine (N-Fen) were studied in great detail. The target compounds were determined by high-performance liquid chromatography with fluorescence detection using 4-(4,5-diphenyl-1H-imidazol-2-yl)benzoyl chloride as a derivatization reagent. After repeated i.p. administration of Fen (5 mg/kg) for 4 days to Zucker rats, shaft and root samples of black and white hair were obtained 1 week after the first administration. It was surprising that Fen and Norf levels in root samples of white hair were much higher than those in shaft or root samples of black hair, strongly suggesting that unknown mechanisms other than the action of melanin take place in the white hair root. Time course profiles for Fen and Norf after administration of a single i.p. dose of Fen or N-Fen were constructed for Zucker and Wistar rats. The percent level of Fen or Norf in white hair was 15–50% of that in black hair at any interval within 600 min after a single administration of Fen in Zucker rats. Even with Wistar rats having only white hair, we could demonstrate the time courses for incorporation of Fen and Norf into white hair. Finally, time course profiles for Fen and Norf were also followed after a single i.p. administration of N-Fen; this experiment showed that the levels of Norf were much higher than those of Fen for both black and white hair samples of Zucker rats at any interval tested.     

Radioimmunoassay of hair for determining opiate-abuse histories

Heroin and morphine metabolites can be detected in hair with the use of commercially available radioimmunoassay reagents and with minor sample preparation. Hair samples obtained from morphine-treated mice and heroin users contained nanogram levels of the drug per milligram of hair (single human hair). The results of the hair analyses for all subjects admitting the use of heroin were positive, whereas the results of only 30% of thin-layer chromatographic urinanalyses of these same subjects were positive. In addition, differences in drug concentration for sections of hair near the scalp and near the distal end correlated with the length of time the drug had been used. These results exemplify the potential advantages of the use of hair analysis over urine and serum analyses in terms of accessibility, sample stability, and long-term retention of information.