In vitro activation of the neuro-transduction mechanism in sensitive organotypic human skin model.
Recent advances in tissue engineering have encouraged researchers to endeavor the production of fully functional three-dimensional (3D) thick human tissues in vitro. Here, we report the fabrication of a fully innervated human skin tissue in vitro that recapitulates and replicates skin sensory function. Previous attempts to innervate in vitro 3D skin models did not demonstrate an effective functionality of the nerve network. In our approach, we initially engineer functional human skin tissue based on fibroblast-generated dermis and differentiated epidermis; then, we promote rat dorsal root ganglion (DRG) neurons axon ingrowth in the de-novo developed tissue. Neurofilaments network infiltrates the entire native dermis extracellular matrix (ECM), as demonstrated by immunofluorescence and second harmonic generation (SHG) imaging. To prove sensing functionality of the tissue, we use topical applications of capsaicin, an agonist of transient receptor protein-vanilloid 1 (TRPV1) channel, and quantify calcium currents resulting from variations of Ca++concentration in DRG neurons innervating our model. Calcium currents generation demonstrates functional cross-talking between dermis and epidermis compartments. Moreover, through a computational fluid dynamic (CFD) analysis, we set fluid dynamic conditions for a non-planar skin equivalent growth, as proof of potential application in creating skin grafts tailored on-demand for in vivo wound shape.
Center for Advanced Biomaterials for HealthCare@CRIB, Istituto Italiano di Tecnologia, Largo Barsanti e Matteucci n. 53, 80125 Naples, Italy.
Francesca
F
Martorina
Interdisciplinary Research Centre on Biomaterials (CRIB), University of Naples Federico II, P.le Tecchio 80, Naples, Italy.
Costantino
C
Casale
Center for Advanced Biomaterials for HealthCare@CRIB, Istituto Italiano di Tecnologia, Largo Barsanti e Matteucci n. 53, 80125 Naples, Italy.
Francesco
F
Urciuolo
Center for Advanced Biomaterials for HealthCare@CRIB, Istituto Italiano di Tecnologia, Largo Barsanti e Matteucci n. 53, 80125 Naples, Italy; Interdisciplinary Research Centre on Biomaterials (CRIB), University of Naples Federico II, P.le Tecchio 80, Naples, Italy; Department of Chemical, Materials and Industrial Production (DICMAPI), University of Naples Federico II, P.le Tecchio 80, Naples, Italy.
Paolo A
PA
Netti
Center for Advanced Biomaterials for HealthCare@CRIB, Istituto Italiano di Tecnologia, Largo Barsanti e Matteucci n. 53, 80125 Naples, Italy. Electronic address: giorgia.imparato@iit.it.
Giorgia
G
Imparato
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D005727:Ganglia, Spinal / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D051381:Rats; D012867:Skin / Q000254:growth & development / Q000294:innervation* / Q000648:ultrastructure; D009435:Synaptic Transmission; D023822:Tissue Engineering / Q000379:methods*
Bioreactor; Endogenous ECM; Innervated skin equivalent; Living prosthetic biomaterial; Organotypic model; Tissue engineering