Constitution of Fibrin-Based Niche for In Vitro Differentiation of Adipose-Derived Mesenchymal Stem Cells to Keratinocytes

Abstract

Epithelialization of chronic cutaneous wound is troublesome and may require use of skin/cell substitutes. Adipose-derived mesenchymal stem cells (ADMSCs) have immense potential as autologous cell source for treating wounds; they can cross the germ layer boundary of differentiation and regenerate skin. When multipotent adult stem cells are considered for skin regeneration, lineage committed keratinocytes may be beneficial to prevent undesirable post-transplantation outcome. This study hypothesized that ADMSCs may be directed to epidermal lineage in vitro on a specifically designed biomimetic and biodegradable niche. Cells were seeded on the test niche constituted with fibrin, fibronectin, gelatin, hyaluronic acid, laminin V, platelet growth factor, and epidermal growth factor in the presence of cell-specific differentiation medium (DM). The ADMSCs grown on bare tissue culture polystyrene surface in DM is designated DM-control and those grown in basal medium (BM) is the BM-control. Lineage commitment was monitored with keratinocyte-specific markers such as cytokeratin 14, cytokeratin 5, cytokeratin 19, and integrin α6 at the transcriptional/translational level. The in vitro designed biomimetic fibrin composite matrix may have potential application as cell transplantation vehicle.

Protein nanocoatings on synthetic polymeric nanofibrous membranes designed as carriers for skin cells

“…[23][24][25] For better regenerative potential and mechanical stability, fibrin matrices have been combined with other biological or synthetic molecules, eg, collagen, 26,27 hyaluronic acid with a cell adhesion-promoting peptide, 28 basic FGF, 29 or EGF. 30 Fibrin has relatively rarely been deposited on supporting substrates, although fibrin selfsupporting matrices are usually fragile. In our previous study, we deposited a fibrin nanocoating on polylactide nanofibrous membranes and observed its positive influence on the behavior of dermal fibroblasts.…”

The use of fat grafting and platelet‐rich plasma for wound healing: A review of the current evidence

“…46,76 When ADSCs are grown in vitro on a scaffold containing adhesion molecules and growth factors found in PRP, they show increased differentiation to keratinocytes suggesting an enhanced benefit to wound healing. 79 It has also been shown in vitro that a majority of platelets remain active at 10 days when cocultured with ADSCs indicating a bilateral prosurvival relationship that may enhance healing potential. 80 PRP has anti-inflammatory properties that may reduce the inflammation and swelling, which encourage degeneration of the fat graft.…”

“…PRP also contains cell adhesion molecules including fibronectin and vitronectin, which help to immobilise growth factors within the fibrin and help the fibrin scaffold act as a matrix for epithelial migration . When ADSCs are grown in vitro on a scaffold containing adhesion molecules and growth factors found in PRP, they show increased differentiation to keratinocytes suggesting an enhanced benefit to wound healing . It has also been shown in vitro that a majority of platelets remain active at 10 days when cocultured with ADSCs indicating a bilateral prosurvival relationship that may enhance healing potential …”

The Use of Adipose-Derived Stem Cells in Selected Skin Diseases (Vitiligo, Alopecia, and Nonhealing Wounds)

“…Human ADSCs can be converted to epithelial cells expressing the characteristics of cytokeratins 5, 14, 19, and α 6; integrins; and even desmoglein 3 [ 60 , 61 ]. They can also differentiate into fibroblast cells, demonstrating not only their morphological similarity but also their ability to also express cell surface proteins including vimentin and fibronectin [ 60 ].…”