Development of a complex, full thickness, in vitro human skin tissue with neuronal and immune components.
Vidal Yucha, Sarah.
2019
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Human skin
equivalents (HSEs) are used for many applications from exploratory research to clinical
needs as wound coverings. HSEs are typically composed of 1-2 cell types in 1-2 tissue
layers without a dermal component. These skin systems miss key components, including
neuronal and immune cells, which are native to the skin and are responsible for numerous
functions including sensation, immune ... read moreand inflammatory responses as part of the
neuro-immuno-cutaneous (NIC) system. The NIC system is also implicated in numerous skin
pathologies including psoriasis, atopic dermatitis, and vitiligo. The goal of this
thesis research was to develop a complex, 3D in vitro HSE that uses primary, human
cells, and captures the NIC system. The complex HSE includes 3 layers: an epidermis,
dermis, and hypodermis; spans 3 systems: cutaneous, neuronal, and immune; and contains
primary human cells including: keratinocytes, fibroblasts, human induced neural stem
cells, and numerous cells from lipoaspirate including adipocytes, endothelial cells, and
tissue-inherent macrophages/monocytes. In addition, it was desirable to incorporate a
biomaterial (silk-collagen composite) matrix that is resistant to contraction to support
long-term studies. This composite matrix preserves the cell-binding domains of collagen
while introducing system stability from the silk component. In the first aim, the
biomaterial matrix (silk-collagen) is evaluated in comparison to the more standard
(collagen), along with the overall construction protocol for the HSE. In the second aim,
several variables are adapted to improve cell characterization and cell-specific effects
are identified in the complex skin tissue model. RNASeq was utilized to assess the HSE
system with respect to the presence or absence of some of the specific tissue layers or
cell types. This analysis determined that inclusion of neuronal and immune components
results in up-regulation of most key pathways of the skin. Overall, this thesis presents
a new 3D, full thickness skin tissue model that includes neuronal and immune components,
consisting of primary human cells. This type of system could ultimately be utilized for
identifying or confirming key biomarkers in the skin, assessing acute and chronic
effects from environmental chemicals and physical exposures, drug uptake, while
simultaneously reducing reliance on animal
studies.
Thesis (Ph.D.)--Tufts University, 2019.
Submitted to the Dept. of Biomedical Engineering.
Advisor: David Kaplan.
Committee: Brian Timko, Kyongbum Lee, and Lloyd Klickstein.
Keyword: Biomedical engineering.read less - ID:
- v692tk39b
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