Abstract
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.
Highlights
Complex tissue models that replicate human biological interactions and the tissues' 3D architecture and function are a requirement for the development of accurate and reliable in vitro systems
We report the use of microscopy-guided laser ablation (MGLA) for the microscale manipulation of a fibroblast-populated collagen hydrogel and the fabrication of microfeatures that enable the recreation of hair follicle (HF) in an organotypic human skin model
Micro-laser ablation allows ablating a selected portion of material delimited by the spot size of the laser
Summary
Complex tissue models that replicate human biological interactions and the tissues' 3D architecture and function are a requirement for the development of accurate and reliable in vitro systems. In the case of skin, currently available models are far from replicating its complexity, representing an oversimplified version composed only by the epidermal and dermal layers. Is an organ endowed with important physiological functions, which are conferred by the presence of specialized cell populations and by functional skin appendages. The hair follicle (HF) critically contributes to the most important physiological functions attributed to skin, including barrier function, thermoregulation, sensory perception, and immunosurveillance. Given the HF high cosmetic value, there is a great deal of interest in prompting HF regenerative therapies, which cannot be dissociated from the development of appropriate test systems representative of the HF formation events, to find novel targeted treatments/drugs
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