Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2014 Oct;3(10):1209-19.
doi: 10.5966/sctm.2013-0217. Epub 2014 Aug 26.

Dermal papilla cells improve the wound healing process and generate hair bud-like structures in grafted skin substitutes using hair follicle stem cells

Gustavo José Leirós  1 , Ana Gabriela Kusinsky  1 , Hugo Drago  1 , Silvia Bossi  1 , Flavio Sturla  1 , María Lía Castellanos  1 , Inés Yolanda Stella  1 , María Eugenia Balañá  2
Affiliations

Dermal papilla cells improve the wound healing process and generate hair bud-like structures in grafted skin substitutes using hair follicle stem cells

Gustavo José Leirós et al. Stem Cells Transl Med. 2014 Oct.

Abstract

Tissue-engineered skin represents a useful strategy for the treatment of deep skin injuries and might contribute to the understanding of skin regeneration. The use of dermal papilla cells (DPCs) as a dermal component in a permanent composite skin with human hair follicle stem cells (HFSCs) was evaluated by studying the tissue-engineered skin architecture, stem cell persistence, hair regeneration, and graft-take in nude mice. A porcine acellular dermal matrix was seeded with HFSCs alone and with HFSCs plus human DPCs or dermal fibroblasts (DFs). In vitro, the presence of DPCs induced a more regular and multilayered stratified epidermis with more basal p63-positive cells and invaginations. The DPC-containing constructs more accurately mimicked the skin architecture by properly stratifying the differentiating HFSCs and developing a well-ordered epithelia that contributed to more closely recapitulate an artificial human skin. This acellular dermal matrix previously repopulated in vitro with HFSCs and DFs or DPCs as the dermal component was grafted in nude mice. The presence of DPCs in the composite substitute not only favored early neovascularization, good assimilation and remodeling after grafting but also contributed to the neovascular network maturation, which might reduce the inflammation process, resulting in a better healing process, with less scarring and wound contraction. Interestingly, only DPC-containing constructs showed embryonic hair bud-like structures with cells of human origin, presence of precursor epithelial cells, and expression of a hair differentiation marker. Although preliminary, these findings have demonstrated the importance of the presence of DPCs for proper skin repair.

Keywords: Adult stem cells; Epidermis; Multipotential differentiation; Skin grafts; Tissue regeneration.

PubMed Disclaimer

Figures

Figure 1.
Figure 1.
Procedure scheme to generate skin constructs using a porcine ADM as scaffold. (A): ADM was seeded with DFs or DPCs. (B): Matrix-side inversion. (C): The ADM was seeded with HFSCs. (D): Some matrices were transferred to Transwell plates and maintained in air-liquid interphase. Abbreviations: ADM, acellular dermal matrix; DF, dermal fibroblast; DPC, dermal papilla cell; HFSC, hair follicle stem cell; PET, polyethylene terephthalate.
Figure 2.
Figure 2.
Histology of skin constructs generated in air-liquid interphase. (A): HFSC. (B): HFSC-DF. (C): HFSC-DPC. Black arrow indicates dyskeratosis; white arrow, parakeratosis. Scale bars = 200 µm (×100 fields) and 50 µm (×400 fields). (D): Bar graph shows the number of epidermal layers from each construct. ∗∗, p < .001. Abbreviations: B st, basal stratum; C st, cornified stratum; DF, dermal fibroblast; DPC, dermal papilla cell; G st, granulosum stratum; HFSC, hair follicle stem cell; S st, spinosum stratum.
Figure 3.
Figure 3.
p63 immunohistochemistry and epidermal invaginations of skin constructs generated in air-liquid interphase. (A): HFSC. (B): HFSC-DF. (C): HFSC-DPC. Scale bars = 20 µm (p63 immunostaining) and 200 µm (epidermal invaginations). (D): Bar graph of p63-positive per total epidermal cells in ×1,000 fields. (E): Bar graph of epidermal invaginations in ×100 fields. White arrows show some p63-positive nucleus and black arrows, some epidermal invaginations. ∗∗, p < .001. Abbreviations: ADM, acellular dermal matrix; DF, dermal fibroblast; DPC, dermal papilla cell; HFSC, hair follicle stem cell.
Figure 4.
Figure 4.
Immunoassay for CD34 neovessel marker in skin constructs grafted in nude mice. The constructs were generated with ADM alone (A), HFSC-DF (B), and HFSC-DPC (C). Scale bars = 50 µm. (D): Bar graph of vessels per dermis surface unit in ×400 fields from each construct. White arrows indicate neovessels below graft epidermis; black arrows, dermal-epidermal detachment. ∗∗, p < .001. Abbreviations: ADM, acellular dermal matrix; DF, dermal fibroblast; DPC, dermal papilla cell; HFSC, hair follicle stem cell.
Figure 5.
Figure 5.
Histology of skin constructs grafted in nude mice. Constructs were generated with ADM alone (A), HFSC-DF (B), and HFSC-DPC (C). Insets: Macroscopic views of the grafts. Black arrows indicate dermal-epidermal detachment; white arrows, epithelial cyst-like structures reminiscent of embryonic hair buds. Scale bars = 200 µm. Abbreviations: ADM, acellular dermal matrix; DF, dermal fibroblast; DPC, dermal papilla cell; EN, epidermis necrosis; GT, granulation tissue; HFSC, hair follicle stem cell.
Figure 6.
Figure 6.
Evaluation of embryonic hair bud-like structures in grafted skin constructs containing HFSC-DPC after 30 days. (A): Hematoxylin and eosin staining. (B): Immunoassay for human leukocyte antigen I. (C): Immunoassay for p63 antigen. (D): Immunoassay for k6hf antigen. Black arrows indicate positive cells for the corresponding antigen; white arrows, the sebaceous glands (unspecifically stained). Abbreviations: DPC, dermal papilla cell; HFSC, hair follicle stem cell.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Papini R. Management of burn injuries of various depths. BMJ. 2004;329:158–160. - PMC - PubMed
    1. Stanton RA, Billmire DA. Skin resurfacing for the burned patient. Clin Plast Surg. 2002;29:29–51. - PubMed
    1. Gallico GG, III, O’Connor NE, Compton CC, et al. Permanent coverage of large burn wounds with autologous cultured human epithelium. N Engl J Med. 1984;311:448–451. - PubMed
    1. Carsin H, Ainaud P, Le Bever H, et al. Cultured epithelial autografts in extensive burn coverage of severely traumatized patients: A five year single-center experience with 30 patients. Burns. 2000;26:379–387. - PubMed
    1. Hafemann B, Ensslen S, Erdmann C, et al. Use of a collagen/elastin-membrane for the tissue engineering of dermis. Burns. 1999;25:373–384. - PubMed

Publication types

  NODES
twitter 2