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. 2022 Jul 29:13:20417314221111868.
doi: 10.1177/20417314221111868. eCollection 2022 Jan-Dec.

Peptide nanogels as a scaffold for fabricating dermal grafts and 3D vascularized skin models

Wafaa T Arab  1 , Hepi H Susapto  1   2 , Dana Alhattab  1   2 , Charlotte A E Hauser  1   2
Affiliations

Peptide nanogels as a scaffold for fabricating dermal grafts and 3D vascularized skin models

Wafaa T Arab et al. J Tissue Eng. .

Abstract

Millions of people worldwide suffer from skin injuries, which create significant problems in their lives and are costly to cure. Tissue engineering is a promising approach that aims to fabricate functional organs using biocompatible scaffolds. We designed ultrashort tetrameric peptides with promising properties required for skin tissue engineering. Our work aimed to test the efficacy of these scaffolds for the fabrication of dermal grafts and 3D vascularized skin tissue models. We found that the direct contact of keratinocytes and fibroblasts enhanced the proliferation of the keratinocytes. Moreover, the expression levels of TGF-β1, b-FGF, IL-6, and IL-1α is correlated with the growth of the fibroblasts and keratinocytes in the co-culture. Furthermore, we successfully produced a 3D vascularized skin co-culture model using these peptide scaffolds. We believe that the described results represent an advancement in the fabrication of skin tissue equivalent, thereby providing the opportunity to rebuild missing, failing, or damaged parts.

Keywords: Ultrashort peptides; fibroblast; keratinocytes; self-assembly; wound healing.

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Conflict of interest statement

Declaration of conflicting interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Figures

Figure 1.
Figure 1.
The morphology of self-assembling IVFK and IVZK peptide hydrogels at a concentration of 4 mg/mL and 3 mg/mL, respectively. The images were taken using scanning electron microscopy (SEM) and transmission electron microscopy (TEM).
Figure 2.
Figure 2.
Bright-field images evidencing the enhancement of keratinocyte proliferation in co-culture with fibroblasts for different scaffolds: Matrigel (a, d, g), IVFK (b, e, h), and IVZK (c, f, i). Scale bars 50 μm.
Figure 3.
Figure 3.
The viability of co-cultured cells on different scaffolds (4 mg/mL IVFK and 3 mg/mL IVZK). Matrigel was used as a positive control (4 mg/mL). Fibroblasts are stained with tracker green fluorescence and keratinocytes with tracker red fluorescence. Significant fibroblast growth on day 7 in Matrigel and IVZK compared to IVFK. On Day 14, a layer of keratinocytes (red fluorescence) has been formed on top of fibroblast (green fluorescence) on IVZK 3D culture compared to IVFK and Matrigel. Scale bars 100 μm.
Figure 4.
Figure 4.
The mitotic activity of fibroblast (green), keratinocytes (red), and nucleus (blue) in 3D co-cultured constructs. The cells were cultured on different scaffolds (4 mg/mL IVFK and 3 mg/mL IVZK), and Matrigel was used as a positive control (4 mg/mL). Scale bars 100 μm.
Figure 5.
Figure 5.
Volume scope scanning electron microscopy images of skin co-culture model after 18 days in the submerged culture at different magnification. Cryosections indicated the formation of different keratinocyte layers (1–3), with cells showing keratohyalin granules (KG) and keratin filaments (KF). Red arrows denoted the collagen secreted from human dermal fibroblast cells (white arrows) within the middle dermis layer.
Figure 6.
Figure 6.
The in vitro release profile of cytokines IL-1α (a), IL-6 (b), and growth factors b-FGF (c) and TGF-β1 (d) in 3D monoculture and co-culture system after 72 h. Fibroblasts and keratinocytes were used for 3D monoculture and co-culture within IVFK and IVZK. Matrigel was used as 3D positive control and TCP (tissue culture plate) as negative 2D control.
Figure 7.
Figure 7.
Fabrication of 3D co-culture vascularized skin constructs using (4 mg/mL IVFK and 3 mg/mL IVZK) Matrigel was used as a positive control (4 mg/mL). This figure shows Z-stack images across the entire depth of vascularized skin constructs. Vimentin, CD31, and CK14 antibodies were used to stain fibroblast, endothelial cells, and keratinocytes, respectively. Scale bars 100 μm.
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