Effect of Living Cellular Sheets on the Angiogenic Potential of Human Microvascular Endothelial Cells

Background: A fundamental issue limiting the efficacy of surgical approaches designed to correct periodontal mucogingival defects is that new tissues rely on limited sources of blood supply from the adjacent recipient bed. Accordingly, therapies based on tissue engineering that leverage local self‐healing potential may represent promising alternatives for the treatment of mucogingival defects by inducing local vascularization. The aim of this study is to evaluate the effect of commercially available living cellular sheets (LCS) on the angiogenic potential of neonatal dermal human microvascular endothelial cells (HMVEC‐dNeo). Methods: The effect of LCS on HMVEC‐dNeo proliferation, migration, capillary tube formation, gene expression, and production of angiogenic factors was evaluated over time. Results: LCS positively influenced HMVEC‐dNeo proliferation and migration. Moreover, HMVEC‐dNeo incubated with LCS showed transcriptional profiles different from those of untreated cells. Whereas increased expression of angiogenic genes predominated early on in response to LCS, late‐phase responses were characterized by up‐ and downregulation of angiostatic and angiogenic genes. However, this trend was not confirmed at the protein level, as LCS induced increased production of most of the angiogenic factors tested (i.e., epidermal growth factor [EGF], heparin‐binding EGF‐like growth factor, interleukin 6, angiopoietin, platelet‐derived growth factor‐BB, placental growth factor, and vascular endothelial growth factor) throughout the investigational period. Finally, although LCS induced HMVEC‐dNeo proliferation, migration, and expression of angiogenic factors, additional factors and environmental pressures are likely to be required to promote the development of complex, mesh‐like vascular structures. Conclusion: LCS favor initial mechanisms that govern angiogenesis but failed to enhance or accelerate HMVEC‐dNeo morphologic transition to complex vascular structures.