title: Continuous supply of TGFβ3 via adenoviral vector promotes type I collagen and viability of fibroblasts in alginate hydrogel.
abstract: In recent years, transforming growth factor-β3 (TGFβ3) has interested more and more researchers with its competence in engineered histogenesis. In the present study we employed recombinant adenoviral vectors to deliver the constitutively active TGFβ3 gene to human dermal fibroblasts, which could maintain the continuous secretion of TGFβ3 from the cells. The expression of type I collagen in the Ad-TGFβ3 group increased significantly in comparison with other three groups: Neg (cells without treatment of the adenovirus), Ad-null (cells with treatment of the adenovirus, without the inserted gene) and Ad-shRNA (cells with treatment of the adenovirus encoding shRNA specific for type I collagen). Additionally, we demonstrated that TGFβ3 enhanced the expression of Smad4 while inhibiting that of MMP-9, thus promoting the collagen transcription via the Smad signal transduction pathway and restraining collagen degradation by MMP-9, which contributed to the increasing type I collagen expression level. As type I collagen mediates cell-material interactions by providing anchorage, the viability of encapsulated fibroblasts in Ad-TGFβ3 group was significantly higher than that in other three groups. Accordingly, this approach forms an effective way to improve the compatibility of non-adhesive hydrogels containing anchorage-dependent cells.
authors.affiliation: Division of Bioengineering, School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore.
authors.forename: Yongchang
authors.initials: Y
authors.lastname: Yao
authors.forename: Feng
authors.initials: F
authors.lastname: Zhang
authors.forename: Ruijie
authors.initials: R
authors.lastname: Zhou
authors.forename: Meng
authors.initials: M
authors.lastname: Li
authors.forename: Dong-An
authors.initials: DA
authors.lastname: Wang
mesh_terms: D000256:Adenoviridae; D000464:Alginates; D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D018914:Cells, Immobilized / Q000166:cytology / Q000378:metabolism; D024042:Collagen Type I / Q000096:biosynthesis* / Q000235:genetics; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D005822:Genetic Vectors; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D006801:Humans; D020100:Hydrogels; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics; D051901:Smad4 Protein / Q000235:genetics / Q000378:metabolism; D013552:Swine; D014158:Transcription, Genetic; D053782:Transforming Growth Factor beta3 / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D000464:Alginates; D024042:Collagen Type I; D006603:Hexuronic Acids; D020100:Hydrogels; C497796:SMAD4 protein, human; D051901:Smad4 Protein; D053782:Transforming Growth Factor beta3; D020723:Glucuronic Acid; D020780:Matrix Metalloproteinase 9