Fabrication of β-Sitosterol-Incorporated Oral/Dental Tissue Scaffolds for Angiogenesis Stimulation

Objectives: β-sitosterol is a phytosterol derived from many plants with a chemical structure similar to cholesterol. This chemical has been demonstrated to have positive angiogenic effects, as well as antioxidant and antimicrobial properties. However, its hydrophobic nature has constrained its application in dental/biomedical constructs. The aim of the present study was to develop an approach to fabricate β-sitosterol-incorporated gelatin scaffolds tailored for effective tissue regeneration.
Methods: To fabricate the scaffolds, β-sitosterol vesicle was first formed using a nonpolar surfactant (TWEEN 20) to address the hydrophobicity challenge. The toxicity of the resulting construct was next evaluated on Human Umbilical Vein Endothelial Cells (HUVECs) using the MTT assay. Subsequently, this vesicle was incorporated in a specific weight ratio (0, 1, 2, and 5%) with 8% wt. gelatin solution. Using a combination of foaming and freeze-drying methods, final samples were produced, and their internal structure, porosity, and mechanical properties were characterized. Furthermore, the incorporation of β-sitosterol in gelatin was investigated through the FTIR method. The values of all tests were expressed as mean ± SD (n = 3).
Results: FTIR analysis demonstrated that β-sitosterol was successfully incorporated in gelatin. The results of the MTT assay indicated that for concentrations less than 0.31 mg/mL, the components were non-toxic for HUVEC. According to the findings, increasing the concentration of β-sitosterol did not have any significant effect on the value of the porosity. Furthermore, the mechanical properties of the samples containing β-sitosterol were comparable to those of gelatin ones. High interconnectivity between the pores was also observed which shows the superior properties of the final products in improving diffusion kinetics.
Conclusions: The method proposed in this study can be used to advance material progress in tissue regeneration in medical and dental settings, particularly in the development of application of cholesterol-like structures, such as β-sitosterol, with promoted angiogenic properties.