title: Engineered dermal equivalent tissue in vitro by assembly of microtissue precursors.
abstract: Tissue-engineered constructs can be fabricated by the assembly of smaller building blocks in order to mimic much of the native biology that is often made from repeating functional units. Our aim was to realize a three-dimensional (3-D) tissue-like construct in vitro by inducing the assembly of functional micrometric tissue precursors (microTPs). MicroTPs were obtained by dynamic cell seeding of bovine fibroblasts on porous gelatine microcarriers using a spinner flask bioreactor. During the dynamic seeding, cells adhered, proliferated and synthesized a thin layer of extracellular matrix (ECM) in and around the macroporous beads, generating the microTPs. The analysis showed that the ECM produced was rich in type I collagen. The cells and ECM layer around the microTPs allowed their biological sintering via cell-cell and cell-matrix interaction after only a few days of dynamic seeding. The assembling ability of microTPs was exploited by placing them in a maturation chamber. After 1 week of culture disc-shaped constructs (1cm in diameter, 1mm in thickness) of completely assembled microTPs were collected. The biohybrid obtained presented both a homogeneous and compact aspect. Moreover, histological and immunohistochemical analyses revealed an abundant ECM, rich in type I collagen, interconnecting the microTPs. The results obtained in this survey pave the way to realizing a 3-D dermal tissue equivalent by means of a bottom-up tissue engineering approach.
authors.affiliation: Interdisciplinary Research Centre on Biomaterials (CRIB), University of Naples Federico II, Naples, Italy.
authors.forename: Carmela
authors.initials: C
authors.lastname: Palmiero
authors.forename: Giorgia
authors.initials: G
authors.lastname: Imparato
authors.forename: Francesco
authors.initials: F
authors.lastname: Urciuolo
authors.forename: Paolo
authors.initials: P
authors.lastname: Netti
mesh_terms: D000818:Animals; D001483:Base Sequence; D019149:Bioreactors; D002417:Cattle; D002478:Cells, Cultured; D017931:DNA Primers; D007150:Immunohistochemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012867:Skin / Q000378:metabolism*; D023822:Tissue Engineering
chemical_list: D017931:DNA Primers