Transplantation of the LGR6+ Epithelial Stem Cell into Full-Thickness Cutaneous Wounds Results in Enhanced Healing, Nascent Hair Follicle Development, and Augmentation of Angiogenic Analytes

Abstract

The LGR6+ epithelial stem cells appear to hold great promise for the development of a clinically useful stem cell–based therapy for the repair of full-thickness wounds and hair regeneration. These results indicate that transplantation of LGR6+ epithelial stem cells promotes epithelialization, hair growth, and angiogenesis in tissues destined for scar formation.

Transplantation of an LGR6+ Epithelial Stem Cell–Enriched Scaffold for Repair of Full-Thickness Soft-Tissue Defects

“…were labeled on the dorsum skin to develop a 2 × 3 grid of 3-mm diameter burn wound beds spaced 1 cm apart as described by Lough et al 3,4 Viability of the LGR6 + green fluorescent protein epithelial stem cells was validated using a IVIS Lumina (PerkinElmer, Akron, Ohio) in vivo bioluminescence imaging system in addition to confocal microscopy for each subgroup at 24 hours. Daily digital imaging was acquired using a Sony PMW-10MD (Sony, Tokyo, Japan) adapted Leica Wild M680 microsurgical scope.…”

“…3,4,[9][10][11][12][13] Previously published literature suggests that successful isolation of the LGR6 + epithelial stem cell from the adnexal follicular bulge can result in a viable and culture-ready stem cell population, which has shown tremendous promise in the development of tissueengineered functional skin constructs. 4,12 In this study, we sought to determine whether the LGR6 + epithelial stem cell could be isolated, expanded in culture, and seeded onto a spectrum of acellular scaffolds to augment healing through the migration, proliferation, and differentiation of the epithelial stem cell into a form of functional skin-capable of producing nascent hair follicles. 3,4 LGR6 + green fluorescent protein epithelial stem cells were obtained from full-thickness C57BL/6(UBC-GFP) murine skin and labeled with fluorescently conjugated antibodies (LGR6, CD34, and CD73) for fluorescence-activated cell sorting and seeded onto a spectrum of 1-cm 2 scaffolding substrates (Fig.…”

“…3,12,13 In addition, cells expressing LGR5 and LGR6 have been shown to promote wound healing, angiogenesis, and nascent hair follicle development in damaged tissues through the modulation of vascular endothelial growth factor (VEGF) and wingless-type MMTV integration site family/β-catenin pathways, suggesting an added role in the coordination of cutaneous healing. 3,4,18,19 Understanding the LGR5 + and LGR6 + epithelial stem cell populations' unique location and regenerative potential, we hypothesize that simple preformed scaffolds, seeded with a self-sustaining LGR6 + epithelial stem cell population, will provide full-thickness wounds with a viable and proliferating stem cell-enriched early tissue substitute. We believe that this enriched hybrid graft system will in turn reinstitute an epithelial stem cell/mesenchymal stem cell interface and potentially increase healing, local tissue polarization, and nascent hair follicle growth.…”

“…14 Recent discovery of LGR5 and LGR6 as extracellular phenotypic defining markers for proliferating adult epithelial stem cells in mammalian skin and intestine permits enhanced clinical utility in the application of the epithelial stem cell in regenerative medicine, wound healing, hair restoration, and biomedical engineering. [3][4][5][15][16][17] These cells, in addition to naturally migrating from the periwound region into defects, have also been found to be capable of providing differentiated tissue elements within both in vitro and in vivo regenerative models. 3,12,13 In addition, cells expressing LGR5 and LGR6 have been shown to promote wound healing, angiogenesis, and nascent hair follicle development in damaged tissues through the modulation of vascular endothelial growth factor (VEGF) and wingless-type MMTV integration site family/β-catenin pathways, suggesting an added role in the coordination of cutaneous healing.…”

“…[4][5][6] Contemporary research has repeatedly validated the importance of both cellular and tissue polarity with respect to fate decisions on stem cell division (symmetric or asymmetric) and for the maintenance of stem cell adhesion, quiescence, and progenitor lineage development. [7][8][9][10] Within the skin, the variety of mesenchymal stem cell populations act as interdependent bioreactors through which the development of local extracellular matrix and the delivery of growth factors promote structural interfaces, leading to eventual tissue architecture and subsequent angiogenesis.…”