The effect of herbal components, pomegranate and icariin on the chondrogenesis of stem cells in the fibrin- micromass hydrogel system

AbstractIntroduction: Tissue engineering has been employed to encourage cartilage renewal using natural or synthetic scaffolds, stem cells, and growth factors. The present study introduces a novel approach based on the chondrogenesis of human adipose-derived mesenchymal stromal cells (ASC)s in the fibrin hydrogel by applying the micro mass culture, using pomegranate and icariin as the herbal components.Materials and Methods: To prepare fibrin hydrogel, a thrombin solution was extracted from human fresh-frozen plasma, and the cryoprecipitate was used as the fibrinogen component following thawing. The ASCs were seeded on the fibrin hydrogel via micro mass culture system. To investigate the chondrogenesis of the ASCs, the cell-seeded hydrogels were cultured in the different culture medias containing TGF-β۳, pomegranate extract, icariin, and the combined medium containing the Icariin and pomegranate.Results: Gene expression measurements and histological staining confirmed that the ASC-seeded hydrogels cultured in the chondrogenic media containing the herbal components could express cartilage-specific genes and synthesize the cartilage-related macromolecules. However, in the presence of TGF-β۳, the hydrogels exhibited a significant expression of these markers.Conclusion: These findings suggest that a combination of these two herbal components could significantly stimulate ASCs and increased cellular proliferation, the expression of chondrogenic genes, the enhanced glycosaminoglycan and collagen synthesis, and the reduction of the hypertrophic phenotypes compared to the TGF-β۳.Keywords: Cartilage tissue engineering, Fibrin hydrogel, Herbal components, Micromass cultureAbstract Introduction: Tissue engineering has been employed to encourage cartilage renewal using natural or synthetic scaffolds, stem cells, and growth factors. The present study introduces a novel approach based on the chondrogenesis of human adipose-derived mesenchymal stromal cells (ASC)s in the fibrin hydrogel by applying the micro mass culture, using pomegranate and icariin as the herbal components. Materials and Methods: To prepare fibrin hydrogel, a thrombin solution was extracted from human fresh-frozen plasma, and the cryoprecipitate was used as the fibrinogen component following thawing. The ASCs were seeded on the fibrin hydrogel via micro mass culture system. To investigate the chondrogenesis of the ASCs, the cell-seeded hydrogels were cultured in the different culture medias containing TGF-β۳, pomegranate extract, icariin, and the combined medium containing the Icariin and pomegranate. Results: Gene expression measurements and histological staining confirmed that the ASC-seeded hydrogels cultured in the chondrogenic media containing the herbal components could express cartilage-specific genes and synthesize the cartilage-related macromolecules. However, in the presence of TGF-β۳, the hydrogels exhibited a significant expression of these markers. Conclusion: These findings suggest that a combination of these two herbal components could significantly stimulate ASCs and increased cellular proliferation, the expression of chondrogenic genes, the enhanced glycosaminoglycan and collagen synthesis, and the reduction of the hypertrophic phenotypes compared to the TGF-β۳. Keywords: Cartilage tissue engineering, Fibrin hydrogel, Herbal components, Micromass culture