In vitro 3D Spheroid Wound Modeling: An Alternative to Experimental Animal Studies

Abstract

ABSTRACT

Laboratory animals have frequently been used in scientific and preclinical pharmaceutical drug safety and efficacy research. Although the introduction of new in silico assays and computer modeling for drug discovery has shown promise in reducing laboratory animal trials, there is still a need to develop in vitro alternatives to in vivo animal models. The in vitro spheroid wound model is one of the best options for developing alternative techniques to animal research as it is the most widely used laboratory animal model. The aim of the study is to using 3D in vitro wound modeling as an alternative to in vivo wound healing assays. In the study, a three-dimensional cell culture (organoid culture) with cell/cell and cell/matrix junctions was generated using the most common Fibroblast and HaCaT cell lines hanging drop technique to replicate the healing stages in the injured skin area. After spheroid epidermal structures were formed, inhibitors and activators were added to the culture medium and their effects on the wound line and 3D cells produced were determined. It was noted that the number of spheroid structures increased significantly and cell-cell interactions became visible in the additional activator groups compared to the control groups. When the inhibitor-treated group was compared with the control groups, it was observed that the formed structures completely disappeared or decreased in amount and cell-cell interactions could not be established. In conclusion, this study offers an alternative to using laboratory animals to evaluate potential medicines and/or extracts in wound healing experiments.