We previously developed a pre-vascularized three-dimensional (3D) cultured human skin model, for which cells were cultured on a porous polymer membrane to supply a culture medium to the tissue. The model was transplanted onto a full-thickness skin defect in a mouse, showing the efficacy as a skin graft. However, there were difficulties in separating the cultured skins from the membranes, as well as in handling the separated soft skins for transplantation. To solve these problems, we recently developed biodegradable porous membranes that enable skin grafting together with the membranes. Poly (lactic-co-glycolic acid) (PLGA) thin films were irradiated with femtosecond laser pulses to create micro through-holes to produce porous membranes. The membrane showed complete decomposition in the mouse subcutaneous tissue within 35 days after implantation. Three-dimensional skins cultured on the membranes were then transplanted together with the membranes onto skin defects in mice, showing reepithelization in the grafted tissues. However, decomposition of the membrane was limited at the early-stage post-transplantation, and insufficient engraftment was observed in some cases. In this study, we increased the hole density of the PLGA membranes to improve their decomposition rate in tissue. We observed that the membrane was completely decomposed in the mouse subcutaneous tissue within 24 days after implantation. On the membranes, 3D skins containing vascular networks were cultivated. However, we encountered another problem that the mechanical strength of the membranes decreased with the increased hole density; some membranes were torn during skin cultivation. Considering this, we plan to further optimize the membrane conditions next.
Three-dimensional (3D) cultured skin with vascular networks is a useful skin substitute that enables rapid perfusion after grafting. However, the efficiency of the medium supply to thick cultured skin is limited, resulting in a reduction in viability. In this study, we applied photobiomodulation (PBM) to control the viability of 3D skins during cultivation. We compared the effects of PBM with illumination by a light-emitting diode (LED) array at four different peak wavelengths (440 nm, 523 nm, 658 nm and 823 nm); PBM was applied once during cultivation and the viability of the 3D skins was evaluated. The results showed that PBM with 823-nm light significantly improved the viability of the skins, while PBM with other wavelengths was not effective. Based on this result, we applied PBM at 823 nm every 24 hours during cultivation, and we found that the viability of the 3D skins with daily PBM was significantly higher than that with single PBM.
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