Morphogenesis of chimeric hair follicles in engineered skin substitutes with human keratinocytes and murine dermal papilla cells

Abstract

Engineered skin substitutes (ESS) have been used successfully to treat life-threatening burns, but lack cutaneous appendages. To address this deficiency, dermal constructs were prepared using collagen-glycosaminoglycan scaffolds populated with murine dermal papilla cells expressing green fluorescent protein (mDPC-GFP), human dermal papilla cells (hDPC) and/or human fibroblasts (hF). Subsequently, human epidermal keratinocytes (hK) or hK genetically modified to overexpress stabilized b-catenin (hK') were used to prepare ESS epithelium. After 10 days incubation at air-liquid interface, ESS were grafted to athymic mice and were evaluated for 6 weeks. Neofollicles were observed in ESS containing mDPC-GFP, but not hDPC or hF, independent of whether or not the hK were genetically modified. Based on detection of GFP fluorescence, mDPC were localized to the dermal papillae of the well-defined follicular structures of grafted ESS. In addition, statistically significant increases in LEF1, WNT10A and WNT10B were found in ESS with neofollicles. These results demonstrate a model for generation of chimeric hair in ESS.

Dermal Papilla Cells Improve the Wound Healing Process and Generate Hair Bud-Like Structures in Grafted Skin Substitutes Using Hair Follicle Stem Cells

“…Sriwiriyanont et al [59] observed neofollicles in grafted engineered skin substitutes with human epidermal keratinocytes and murine dermal papilla cells but not when the dermal cells were of human origin. In other study [60], they grafted in nude mice, chimeric populations of cultured human keratinocytes from neonatal foreskins and cultured murine dermal papilla cells from adult green fluorescent protein transgenic mice.…”

Hair miR‐29a levels are decreased in patients with scleroderma

“…Recently, Lefkowitz et al demonstrated the presence of miRNAs in the hair shafts although they do not have nuclear components (7,8). We also reported that the hair miRNA levels did not correlate with serum miRNA levels (9), suggesting that they can be independent biomarkers.…”

A new COL3A1 mutation in Ehlers–Danlos syndrome type IV

“…Peripheral artery occlusive disease is an uncommon symptom in EDS IV, although fragility of the large vessels and easy bruising are well documented (3)(4)(5)(6)11,12). In an earlier report using intravital capillaroscopy and fluorescence videomicroscopy, SupertiFurga et al (13) described microangiopathy of the skin capillaries, with microbleedings and microaneurysms indicating that EDS IV might not only be restricted to the large vessels.…”

“…The clinical diagnosis of EDS IV is made according to the 1997 Villefranche nosology defining major and minor diagnostic criteria (5). The underlying defect is a mutation in the COL3A1 gene located on the long arm of chromosome 2, in ª 2013 John Wiley & Sons A/S Experimental Dermatology, 2013, 22, 216-238 position 2q24.3-q31 that encodes the pro-a1 chain of type III collagen (3)(4)(5)(6). The resulting impaired function or reduced synthesis of type III collagen leads to highly increased fragility of connective tissues resulting in arterial tears or dissections and uterine and/or bowel ruptures as potentially fatal complications.…”