In vitro models to study angiogenic effects of dermal papilla cells as cellular component of skin substitutes

OPPENHEIMER, FLORENCIA; CERUTI, JULIETA MARÍA; LEIRÓS, GUSTAVO JOSÉ; BALAÑA, MARÍA EUGENIA

VIRTUAL

Congreso; European Society for Dermatological Research Annual Meeting; 2021

European Society for Dermatological Research

Tissue-engineered skin represents a useful strategy for the treatment of deep skin injuries.However, its correct vascularization remains a major challenge. We have shown that thepresence of dermal papilla cells (DPCs) in these constructs favors the vascularization process,resulting in a better wound healing and graft take. We also have seen DPC-spheres cultureincrease the expression of angiogenic genes as VEGF, angiogenin and FGF. Since angiogenesis in scaffolds is essential for grafts to survive and integrate with existing host tissue, ouraim was to develop two in vitro models to quantify angiogenesis stimulation of monolayerand spheres [G1] cultures of DPC. The first model was a migration assay, in which DPC ordermal fibroblasts (DF) were co-cultured as monolayers with Human Umbilical Vein Endothelial Cells (HUVEC) in a transwell system. We observed more migrated HUVEC in thecoculture with DPC than in the DF or medium controls. The second model was an endothelialcell tube formation assay where HUVEC, resuspended in DPC-conditioned medium andseeded on Matrigel, showed an increase in the total tubule’s length and in the number ofsegments and joints, compared to the medium control. To be able to compare the inductivemolecules secretion between monolayer and spheres [G2] -DPC cultures, we looked for thecondition in which both systems contained the same amount of metabolically active cells. Inour culture conditions, 55 spheres [G3] were metabolically equivalent to 104 cells/cm2monolayer seeded cells. Using this equivalence, we are evaluating the effects that sphereculture conditions have on the angiogenic abilities of DPCs. We conclude that these twoangiogenic models are useful to measure, in vitro, the stimulation of angiogenesis of cellcultures to be used as dermal components in skin substitutes. In this way, the use of DPC inskin substitutes could favor the vascularization of the grafts, and thus the closure of the woundand the graft take.