Abstract

An important challenge in the fabrication of tissue engineered constructs for regenerative medical applications is the development of processes capable of delivering cells and biomaterials to specific locations in a consistent manner. Electrospraying live cells has been introduced in recent years as a cell seeding method, but its effect on phenotype nor genotype has not been explored. A promising candidate for the cellular component of these constructs are human adipose-derived stem cells (hASCs), which are multipotent stem cells that can be differentiated into fat, bone, and cartilage cells. They can be easily and safely obtained from adipose tissue, regardless of the age and sex of the donor. Moreover, these cells can be maintained and expanded in culture for long periods of time without losing their differentiation capacity. In this study, hASCs directly incorporated into a polymer solution were electrosprayed, inducing differentiation into chondrocytes, without the addition of any exogenous factors. Multiple studies have demonstrated the effects of exposing hASCs to biomolecules—such as soluble growth factors, chemokines, and morphogens—to induce chondrogenesis. Transforming growth factors (e.g., TGF-β) and bone morphogenetic proteins are particularly known to play essential roles in the induction of chondrogenesis. Although growth factors have great therapeutic potential for cell-based cartilage regeneration, these growth factor-based therapies have presented several clinical complications, including high dose requirements, low half-life, protein instability, higher costs, and adverse effects in vivo. The present data suggests that electrospraying has great potential as hASCs-based therapy for cartilage regeneration.

Highlights

  • An important challenge in the fabrication of tissue engineered constructs for regenerative medical applications is the development of processes capable of delivering cells and biomaterials to specific locations in a consistent manner

  • The flow rate was set as 200 μL/min, and the high voltages of 10 and 15 kV direct current (DC) were applied to the nozzle

  • Experiences several forces simultaneously: the electric force Fe ∼ ε0φ2 where ε0 is the electrical permittivity of vacuum, the downward gravitational force is represented by Fg ∼ ρDo3g, the downward inertial force caused by the momentum per unit time injected by the flow into the meniscus is represented by P ∼ ρQ2/Di2, and the surface tension in the opposite direction is indicated by Fγ ∼ Doγ

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Summary

Introduction

An important challenge in the fabrication of tissue engineered constructs for regenerative medical applications is the development of processes capable of delivering cells and biomaterials to specific locations in a consistent manner. A promising candidate for the cellular component of these constructs are human adipose-derived stem cells (hASCs), which are multipotent stem cells that can be differentiated into fat, bone, and cartilage cells They can be and safely obtained from adipose tissue, regardless of the age and sex of the donor. While high precision techniques exist, such as bioprinting for the fabrication of 3D tissue structures via deposition of a bioink composed of cells and b­ iomaterials[2], none of these techniques can provide cellular stimulation in the way electrospraying can This is an electrohydrodynamic process, like electrospinning, which involves the ejection of a stream of polymer solution or “melt” using an electrically charged jet, which can create fine droplets of varying size, depending on the strength of the electric field used. This work is one of the first to study the specific effects of electrospraying on cell differentiation; it builds on prior work demonstrating that cells can survive electrospraying, and will advance a new approach of cell differentiation in tissue engineering

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