■促进伤口的快速愈合以及所有皮肤附件的功能恢复是再生医学的主要目标。到目前为止,目前的方法,包括常用的背部切除伤口模型(BEWM)和脚爪皮肤烫伤模型,专注于评估毛囊(HF)或汗腺(SwG)的再生。如何通过对HFs的同步评估来实现从头附件的再生,SwG和皮脂腺(SeG)仍然具有挑战性。这里,我们开发了一种掌侧皮肤切除伤口模型(VEWM),适用于检查多附件修复的皮肤伤口愈合,以及神经支配,为皮肤伤口的完美再生提供了新的研究范式。
■宏观观察,碘-淀粉试验,形态学染色和qRT-PCR分析用于检测HFs的存在,SwGs,SeG和掌侧皮肤中神经纤维的分布。伤口愈合过程监测,HE/Masson染色,进行分形分析和行为反应评估,以验证VEWM可以模仿人类瘢痕形成和感觉功能损害的病理过程和结果。
■HF限于脚垫间。SwGs密集分布在脚垫中,分散在IFP中。掌侧皮肤有丰富的神经支配。术后1、3、7、10天VEWM创面面积分别为89.17%±2.52%,71.72%±3.79%,55.09%±4.94%和35.74%±4.05%,最终瘢痕面积占初始伤口的47.80%±6.22%。术后1、3、7、10天BEWM创面面积分别为61.94%±5.34%,51.26%±4.89%,12.63%±2.86%和6.14%±2.84%,最终瘢痕面积占初始伤口的4.33%±2.67%。对VEWM与人的创伤后修复部位进行了分形分析:空洞值,0.040±0.012vs0.038±0.014;分形维数值,1.870±0.237对1.903±0.163。评估了正常皮肤与创伤后修复部位的感觉神经功能:机械阈值,1.05±0.52vs4.90g±0.80;针刺反应率,100%vs71.67%±19.92%,和温度阈值,50.34°C±3.11°Cvs52.13°C±3.54°C。
■VEWM紧密反映了人体伤口愈合的病理特征,可用于皮肤多附件再生和神经支配评估。
UNASSIGNED: Promoting rapid wound healing with functional recovery of all skin appendages is the main goal of regenerative medicine. So far current methodologies, including the commonly used back excisional wound model (BEWM) and paw skin scald wound model, are focused on assessing the regeneration of either hair follicles (HFs) or sweat glands (SwGs). How to achieve de novo appendage regeneration by synchronized evaluation of HFs, SwGs and sebaceous glands (SeGs) is still challenging. Here, we developed a volar skin excisional wound model (VEWM) that is suitable for examining cutaneous wound healing with multiple-appendage restoration, as well as innervation, providing a new research paradigm for the perfect regeneration of skin wounds.
UNASSIGNED: Macroscopic observation, iodine-starch test, morphological staining and qRT-PCR analysis were used to detect the existence of HFs, SwGs, SeGs and distribution of nerve fibres in the volar skin. Wound healing process monitoring, HE/Masson staining, fractal analysis and behavioral response assessment were performed to verify that VEWM could mimic the pathological process and outcomes of human scar formation and sensory function impairment.
UNASSIGNED: HFs are limited to the inter-footpads. SwGs are densely distributed in the footpads, scattered in the IFPs. The volar skin is richly innervated. The wound area of the VEWM at 1, 3, 7 and 10 days after the operation is respectively 89.17% ± 2.52%, 71.72% ± 3.79%, 55.09 % ± 4.94% and 35.74% ± 4.05%, and the final scar area accounts for 47.80% ± 6.22% of the initial wound. While the wound area of BEWM at 1, 3, 7 and 10 days after the operation are respectively 61.94% ± 5.34%, 51.26% ± 4.89%, 12.63% ± 2.86% and 6.14% ± 2.84%, and the final scar area accounts for 4.33% ± 2.67% of the initial wound. Fractal analysis of the post-traumatic repair site for VEWM vs human was performed: lacunarity values, 0.040 ± 0.012 vs 0.038 ± 0.014; fractal dimension values, 1.870 ± 0.237 vs 1.903 ± 0.163. Sensory nerve function of normal skin vs post-traumatic repair site was assessed: mechanical threshold, 1.05 ± 0.52 vs 4.90 g ± 0.80; response rate to pinprick, 100% vs 71.67% ± 19.92%, and temperature threshold, 50.34°C ± 3.11°C vs 52.13°C ± 3.54°C.
UNASSIGNED: VEWM closely reflects the pathological features of human wound healing and can be applied for skin multiple-appendages regeneration and innervation evaluation.