Engineering complex tissues requires the use of advanced biofabrication techniques that allow the replication of the tissue's 3D microenvironment, architecture and cellular interactions. In the case of skin, the most successful strategies to introduce the complexity of hair follicle (HF) appendages have highlighted the importance of facilitating direct interaction between dermal papilla (DP) cells and keratinocytes (KCs) in organotypic skin models. In this work, we took advantage of microscopy‐guided laser ablation (MGLA) to microfabricate a fibroblast‐populated collagen hydrogel and create a subcompartment that guides the migration of KCs and lead their interaction with DP cells to recreate follicular structures. Upon definition of the processing parameters (laser incidence area and power), MGLA was used to create 3D microchannels from the surface of a standard organotypic human skin model up to the aggregates containing DP cells and KCs, previously incorporated into the dermal‐like fibroblast‐collagen layer. Analysis of the constructs showed that the fabricated microfeatures successfully guided the fusion between epidermal and aggregates keratinocytes, which differentiated into follicular‐like structures within the organotypic human skin model, increasing its functionality. In summary, we demonstrate the fabrication of a highly structured 3D hydrogel‐based construct using MGLA to attain a complex skin model bearing folliculoid structures, highlighting its potential use as an in vitro platform to study the mechanisms controlling HF development or for the screening of bioactive substances.

中文翻译：

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显微镜引导下的激光消融术可创建具有卵泡状附件的复杂皮肤模型

工程复杂组织需要使用先进的生物制造技术，该技术可以复制组织的3D微环境，架构和细胞相互作用。就皮肤而言，引入毛囊（HF）附件复杂性的最成功策略突出了在器官型皮肤模型中促进真皮乳头（DP）细胞和角质形成细胞（KC）之间直接相互作用的重要性。在这项工作中，我们利用了显微镜引导的激光消融（MGLA）来微制造成纤维细胞填充的胶原水凝胶，并创建了一个小隔间，该小隔间可指导KC的迁移并引导它们与DP细胞的相互作用，从而重建卵泡结构。定义处理参数（激光入射面积和功率）后，MGLA用于创建从标准器官型人类皮肤模型的表面到包含DP细胞和KC的聚集体的3D微通道，这些聚集体先前已合并到真皮样成纤维细胞胶原蛋白层中。对构建体的分析表明，所制造的微观特征成功地指导了表皮和聚集的角质形成细胞之间的融合，该融合作用在器官型人皮肤模型内分化为卵泡状结构，从而增强了其功能性。总而言之，我们演示了使用MGLA制备高度结构化的基于3D水凝胶的构建体，从而获得带有卵泡结构的复杂皮肤模型，强调了其潜在用途，可作为体外平台来研究控制HF发育的机制或用于筛选生物活性物质。物质。