title: Growth Factors Cross-Linked to Collagen Microcarriers Promote Expansion and Chondrogenic Differentiation of Human Mesenchymal Stem Cells.
abstract: Tissue engineering is a field in progressive expansion and requires constant updates in methods and devices. One of the central fields is the development of biocompatible, biodegradable, and injectable scaffolds, such as collagen microcarriers. To enhance cell attachment and produce a cost-effective cell culture solution with local stimulation of cells, basic fibroblast growth factor (bFGF) or transforming growth factor-β1 (TGF-β1) was covalently immobilized on microcarriers either by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide (EDC/NHS) or riboflavin/UV (RB/UV) light-mediated cross-linking. Collagen microcarriers cross-linked with bFGF or TGF-β1 were used for expansion and chondrogenic differentiation of human mesenchymal stem cells (MSCs). Evaluation methods included cell viability test, chondrogenic marker expression (aggrecan and collagen type I and type II), histological detection of proteoglycans, and immunohistochemical analysis. Cross-linking strengthened the collagen structure of the microcarriers and reduced collagenase-mediated degradation. MSCs effectively proliferated on microcarriers cross-linked with bFGF, especially by EDC/NHS cross-linking. Chondrogenic differentiation of MSCs was induced by TGF-β1 cross-linked on microcarriers, promoting gene expression and protein accumulation of aggrecan and collagen type I and type II, as well as proteoglycans. Cross-linking by RB/UV enhanced chondrogenesis more than any other group. In addition, cross-linking reduced scaffold shrinkage exerted by MSCs during chondrogenesis, a desirable feature for microcarriers if used as tissue defect filler. In conclusion, cross-linking of bFGF or TGF-β1 to collagen microcarriers supported in vitro proliferation and chondrogenesis, respectively. If translated in vivo and in clinical practice, such approach might lead a step closer to development of a cost-effective and locally acting device for cell-based therapy.
authors.affiliation: 1 Biomedical Laboratories, Swiss Paraplegic Research , Nottwil, Switzerland .
authors.forename: Alessandro
authors.initials: A
authors.lastname: Bertolo
authors.affiliation: 1 Biomedical Laboratories, Swiss Paraplegic Research , Nottwil, Switzerland .
authors.forename: Fanny
authors.initials: F
authors.lastname: Arcolino
authors.affiliation: 1 Biomedical Laboratories, Swiss Paraplegic Research , Nottwil, Switzerland .
authors.forename: Simona
authors.initials: S
authors.lastname: Capossela
authors.affiliation: 2 Center for High Instruments, Tuscia University , Viterbo, Italy .
authors.forename: Anna Rita
authors.initials: AR
authors.lastname: Taddei
authors.affiliation: 3 Cantonal Hospital of Lucerne , Lucerne, Switzerland .|4 Swiss Paraplegic Centre , Nottwil, Switzerland .
authors.forename: Martin
authors.initials: M
authors.lastname: Baur
authors.affiliation: 4 Swiss Paraplegic Centre , Nottwil, Switzerland .
authors.forename: Tobias
authors.initials: T
authors.lastname: Pötzel
authors.affiliation: 1 Biomedical Laboratories, Swiss Paraplegic Research , Nottwil, Switzerland .|5 Institute for Surgical Technology and Biomechanics, University of Bern , Bern, Switzerland .
authors.forename: Jivko
authors.initials: J
authors.lastname: Stoyanov
mesh_terms: D000328:Adult; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D020219:Chondrogenesis / Q000187:drug effects*; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D008875:Middle Aged; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry; D053773:Transforming Growth Factor beta1 / Q000737:chemistry / Q000494:pharmacology
chemical_list: D053773:Transforming Growth Factor beta1; D016222:Fibroblast Growth Factor 2