Biomimetic Bilayered Gelatin‐Chondroitin 6 Sulfate‐Hyaluronic Acid Biopolymer as a Scaffold for Skin Equivalent Tissue Engineering

Abstract

In order to develop an adequate scaffold for skin tissue engineering, a bilayered gelatin-chondroitin 6 sulfate-hyaluronic acid membrane with a different pore size on either side was prepared. A rete ridges-like topographic microporous structure, which provided the paracrine crosstalk in the epithelial-mesenchymal interactions, was formed. Chondroitin-6-sulfate and hyaluronic acid were incorporated within the gelatin membrane to mimic skin composition and create an appropriate microenvironment for cell proliferation, differentiation, and migration. In the study, the lower layer of the membrane (pore size: 150 microm) was seeded with dermal fibroblasts and acted as the feeder layer for keratinocyte inoculation. Meanwhile, the upper layer (pore size: 20-50 microm) was seeded with keratinocytes for epidermalization. The dermal fibroblasts were dynamically seeded in a self-designed spinner flask for more even cell distribution. The keratinocytes were cultured in submerged conditions for 5 days and then in an air-liquid interface condition for further differentiation. After being cultured for 21 days, the upper layer, seeded with keratinocytes, developed into an epidermis-like structure while the lower part, which was seeded with dermal fibroblasts developed into a dermis-like structure. A histological examination and immunostain were used to prove that keratinocytes maintain their phenotype and stratified epidermis layers were formed within 21 days. In brief, the bilayered skin substitute with biological dermal analog and epidermal structure was successfully fabricated. From this study, we can suggest that the culture model is suitable for autologous skin equivalent preparation.

The Effect of Sterilization Methods on the Physical Properties of Silk Sericin Scaffolds

“…The SEM and pore size measurements indicate that the ethanol treatment significantly reduced the pore size of the sericin scaffolds. However, the pore sizes of all of the sericin scaffolds in this study still remained within the range of 20-50 μm, which is suitable for the proliferation of keratinocytes (29).…”

Fabrication of gelatin–hyaluronic acid hybrid scaffolds with tunable porous structures for soft tissue engineering

“…HA was also demonstrated to have an important role in promoting the penetration of cells into scaffolds due to its high water-holding capacity and intrinsic swelling property in biological media [7]. As a consequence, HA and its derivatives have been extensively explored as scaffolding materials for a variety of tissue engineering applications including skin [8][9][10], cartilage [11][12][13], bone [14], ligament [15], brain [16], and nerve [17,18]. However, the glucuronic acid residues of HA contain a carboxyl group which is negatively charged at physiological pH and ionic strength.…”

Bioengineering the Skin–Implant Interface: The Use of Regenerative Therapies in Implanted Devices

“…65 HA is charged, but the only non-sulfated glycosaminoglycan. For this reason, some investigators have been using scaffolds containing chondroitins, such as chondroitin-6-sulfate in skin applications 155 to combine structural characteristics with charge and sulfation. In addition, analogs of extracellular matrix containing chondroitins inhibit scar synthesis by inducing partial regeneration of the dermis and the epidermis.…”

“…83 During our previous discussion of hyaluronic acid, we have already indicated the main beneficial effects of chondroitin. It can provide scaffolding and support, 154 can play a role in the reconstruction of the cellular glycocalyx, 51 combine structural characteristics with charge and sulfation, 155 and may help in reducing or inhibit scars by inducing partial regeneration of the dermis and the epidermis. 129 A possible issue in the use of chondroitin is currently their commercial presentation, which comes from a variety of sources.…”