The tissue-engineered epidermal (TEE), composed of biocompatible vectors and autogenous functional cells, is a novel strategy to solve the problem of shortage of donor skin sources. The human primary keratinocyte (HPK), the major skin components, are self-evident vital in wound healing and was considered as one of the preferred seed cells for TEEs. Since the process of separating HPKs from the skin triggers a stress state of the cells, achieving its rapid adhesion and proliferation on biomaterials remains challenging. The key to the clinical application is to ensure the normal function of cells while improving the proliferation ability in vitro, and to complete the complex mesenchymal epithelialization to achieve tissue remodeling after vivo implantation. Herein, in order to aid HPKs adhesion and proliferation in vitro and promoting wound healing, we developed a three dimensional collagen scaffold with Y-27632 sustainedly released from the nanoplatform, hollow mesoporous organosilica nanoparticles (HMON). The results showed that the porous structure within the TEE supports the implanted HPKs expanding in a three-dimensional mode to jointly construct the tissue-engineered epidermis in vitro and inhibited the mitochondria-mediated cell apoptosis. It was confirmed that the TEEs with suitable degradation rate could maintain drug release after implantation and could accelerate vascularization of wound base and further revealed the involvement of mesenchymal transformation of transplanted HPKs during skin regeneration in a nude mouse model with full-thickness skin resection. In conclusion, our study highlights the great potential of constructing TEE using a nanoparticle platform for the treatment of large-area skin defects.
BACKGROUND AND PURPOSE: Chemical 2, 4-dinitrofluorobenzene (DNFB),
commonly called as Sanger’s reagent, is well known as skin sensitizer to
cause dermatitis. However, how the DNFB causes skin inflammation remains
unknown. In this study we aimed at identifying the molecular target that
DNFB acts on. EXPERIMENTAL APPROACH: We used a fluorescent calcium
imaging plate reader as an initial screening assay and patch-clamp
recordings for validation. Molecular docking in combination with
site-directed mutagenesis was carried out to investigate DNFB binding
sites in TRPA1 ion channel. KEY RESULTS: We found the chemical DNFB that
selectively activates TRPA1 channel with EC50 of 2.36 ± 0.26 µM.
Single-channel recording reveals that DNFB increases the channel open
probability and acts on three residues C621, Y658 and E625 critical for
DNFB-mediated TRPA1 activation. CONCLUSION AND IMPLICATIONS: Our
findings not only explain a molecular mechanism underlying the
dermatitis and pruritus caused by chemical DNFB, but also provides a
molecular tool that is 7.5-time more potent than current AITC molecule
and can be used for elucidating TRPA1 channel pharmacology and
pathology.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.