在储存于可防止毛囊细胞凋亡的缓冲液中的毛囊中，体外毛干伸长得到增强。

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

作者信息

Krugluger Walter, Moser Karl, Moser Claudia, Laciak Katharina, Hugeneck Joerg

机构信息

Moser Medical Group, Clinic Vienna, Vienna, Austria.

出版信息

Dermatol Surg. 2004 Jan;30(1):1-5; discussion 5. doi: 10.1111/j.1524-4725.2004.30010.x.

DOI:10.1111/j.1524-4725.2004.30010.x

PMID:14692918

Abstract

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.

摘要

背景

毛囊移植过程中储存的微小移植物的活力和存活是微小移植物移植手术的重要限制因素。在本研究中，我们通过测量体外毛干伸长（HSE）5天，研究了不同储存溶液和凋亡细胞死亡（ACD）抑制剂对毛囊细胞活力的影响。

方法

从接受常规微小移植物移植的知情患者获取的微小移植物，在室温下于磷酸盐缓冲盐溶液（PBS）或含有不同浓度ACD抑制剂氨基胍（AMG）、激素（胰岛素、氢化可的松）、14,15-环氧-二十碳三烯酸（14,15-EET）或它们组合的HEPES缓冲的杜氏改良 Eagle培养基（DMEM）中储存5小时。

结果

在体外，与PBS相比，储存在DMEM中的微小移植物的HSE显著增加（2.3%±0.6%对28.4%±3.9%，P<0.0001）。添加AMG（10微克/毫升）或14,15-EET（1纳克/毫升）的DMEM进一步增加了体外HSE（分别为33.9%±7.1%，p = 0.01和32.8%±6.1%，P = 0.02）。通过测定细胞质组蛋白相关DNA片段对储存的微小移植物中的ACD进行评估，证实了HSE的结果。在含血清培养基中培养36小时后可检测到ACD，与储存在DMEM中的微小移植物相比，储存在PBS中的微小移植物中的ACD更高（A405nm/A492nm：分别为1.63±0.21对1.42±0.07；P<0.01）。添加AMG进一步降低了微小移植物中血清诱导的ACD（DMEM为1.42±0.07对DMEM/AMG为0.90±0.11，P<0.0001）。