Gelatine–collagen photo-crosslinkable 3D matrixes for skin regeneration

Abstract

Immediate care of skin wounds and burns is essential to repair this mechanical and chemical barrier to infections. Hydrogels have become one of the standard methods for wound care. Here, gelatine–collagen photo-crosslinkable matrixes or hydrogels were manufactured by two-photon polymerization (TPP) or one-photon UV exposure using a Digital Light Processing (DLP) setup. Both techniques are able to construct matrixes from computer-aided design models, which is important for future clinical applications in which wound dressings should be customized. Although TPP can mimic the 3D dermo-epidermal junction with a high spatial resolution (i.e., ∼6 μm³), the manufacturing time was too slow to produce large wound dressings. Therefore, a DLP setup was explored in this study to fabricate large 2D matrixes of several cm² using the same photo-resist as for TPP, except for the photoinitiator. The fibroblast viability, adherence, and proliferation were analysed in time on both 3D and 2D matrixes in vitro using two-photon microscopy. For both types of matrixes, the adherence and proliferation of fibroblasts (3T3-NIH) were optimal for stiff structures with a Young's modulus of 191 ± 35 kPa compared to softer matrixes of 37 ± 12 kPa. Fibroblast showed complete confluence on Day 14 after seeding on these matrixes, which may create the granulation tissue composed of fibronectin, collagen, and various proteoglycans in the future dermis before repair of the epidermis and disintegrating of their host matrix. For the monitoring of this repair, gelatine–collagen matrixes can easily incorporate bio-optical sensors for the simultaneous monitoring of inflammation processes and wound healing in time.