Abstract
BackgroundDespite the significant progress in the development of skin equivalents (SEs), the problem of noninvasively assessing the quality of the cell components and the collagen structure of living SEs both before and after transplantation remains. Undoubted preference is given to in vivo methods of noninvasive, label-free monitoring of the state of the SEs. Optical bioimaging methods, such as cross-polarization optical coherence tomography (CP OCT), multiphoton tomography (MPT), and fluorescence lifetime imaging microscopy (FLIM), present particular advantages for the visualization of such SEs.MethodsIn this study, we simultaneously applied several visualization techniques for skin model examination. We investigated the structure and quality of dermal equivalents containing dermal papilla (DP) cells and dermal fibroblasts (FBs) using CP OCT, MPT, and FLIM. Both the energy metabolism of the cell components and the structuring of the collagen fibrils were addressed.ResultsBased on the data from the fluorescence lifetimes and the contributions of protein-bound NAD(P)H, a bias toward oxidative metabolism was indicated, for the first time, in both the DP cells and FBs on day 14 of SE cultivation. The CP OCT and MPT data also indicated that both DP cells and FBs structured the collagen gel in a similar manner.ConclusionIn this study, multimodal label-free imaging of the structure and quality of living dermal equivalents was implemented for the first time with the use CP OCT, MPT, and FLIM of NAD(P)H. Our data suggest that the combination of different imaging techniques provides an integrated approach to data acquisition regarding the structure and quality of dermal equivalents, minimizes the potential disadvantages of using a single method, and provides an ideal information profile for clinical and research applications.
Highlights
Despite the significant progress in the development of skin equivalents (SEs), the problem of noninvasively assessing the quality of the cell components and the collagen structure of living Skin equivalent (SE) both before and after transplantation remains
Vimentin was expressed in both the dermal papilla (DP) cells and Dermal fibroblast (FB) for the entire culture period (14 days). α-Smooth muscle actin (SMA) was detected in both cell types only on the final day (14) of cultivation
Versican at the confidence level was detected in the DP cells at all stages (Fig. 1)
Summary
Despite the significant progress in the development of skin equivalents (SEs), the problem of noninvasively assessing the quality of the cell components and the collagen structure of living SEs both before and after transplantation remains. The main directions of such developments are in the isolation and cultivation of cell cultures in vitro, The result of work in these areas is the creation of histotypical or functional analogs (equivalents) of tissues and organs, in particular human skin equivalents. Skin equivalents (SEs) are already used clinically to promote the healing of acute and chronic wounds or in pharmaceutical research as test systems [3]. Human SEs are bioengineered structures (skin substitutes) consisting of cell components, i.e., cultured human. Tissue-engineering structures using stem cells (SCs) have been developed [5] over the same period. Such development of methods for treating injuries and wound healing involves mainly adult SCs, multipotent mesenchymal stromal cells (MSCs) [6]
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