Engineering a Multilayered Skin Substitute with Keratinocytes, Fibroblasts, Adipose Derived Stem Cells and Adipocytes

Area of Science:

• Tissue Engineering
• Regenerative Medicine
• Biomaterials Science

Background:

• Current skin substitutes primarily focus on dermal and epidermal layers, neglecting the hypodermis.
• The hypodermis plays a crucial role in skin structure, insulation, and energy storage.
• A comprehensive skin substitute requires regeneration of all three layers: epidermis, dermis, and hypodermis.

Purpose of the Study:

• To develop a functional, three-dimensional skin substitute incorporating a hypodermal layer.
• To utilize fibrin sealant as a hydrogel scaffold for skin tissue engineering.
• To investigate the potential of adipose-derived stem cells (ASCs) and mature adipocytes for hypodermal regeneration.

Main Methods:

• A three-layered construct was fabricated using fibrin hydrogel.
• The hypodermal layer was created by seeding ASCs and mature adipocytes within fibrin.
• The dermal layer was formed with fibroblasts in fibrin, and the epidermal layer with keratinocytes using air-liquid interface culture.

Main Results:

• A viable three-layered skin substitute was successfully generated.
• The construct incorporated distinct epidermal, dermal, and hypodermal layers.
• Cultivation under air-liquid interface conditions supported keratinocyte differentiation.

Conclusions:

• Fibrin hydrogel is a suitable scaffold for creating multi-layered skin substitutes.
• The developed construct shows promise for regenerating the hypodermis, dermis, and epidermis.
• This approach offers a potential solution for complex skin wound healing and reconstruction.