Interactive effects of growth factors and three-dimensional scaffolds on multipotent mesenchymal stromal cells.
The creation of tissue-engineered constructs with autologous cells is a central goal in regenerative medicine. With respect to ligament replacement, we have evaluated the influences of matrix and growth factors on hMSCs (human mesenchymal stromal cells). hMSCs were seeded in two different 3D (three-dimensional) systems consisting of either a collagen type I gel or a synthetic PLA [poly-(L-lactic acid)] scaffold. After cultivation for 14 days with rhTGFbeta1 (recombinant human transforming growth factor beta1), rhPDGF-BB (recombinant human platelet-derived growth factor homodimer of B-chain) or rhBMP13 (recombinant human bone morphogenetic protein 13), we assessed the proliferation potential, mRNA expression and protein expression of various matrix-interacting and matrix-degrading molecules by quantitative real-time RT (reverse transcriptase)-PCR, immunohistochemistry and gelatin zymography in comparison with unstimulated cells. Cellular reactions to the type of scaffold or soluble factors could be found in the expression of tenascin-C as well as integrin subunits alpha1, alpha3 and beta1. Collagen type X expression was induced by 3D culture and stimulated by rhTGFbeta1 on PLA. The expression of MMP-1 (matrix metalloproteinase 1) tended to increase, and MMP-13 was induced in the collagen culture system. The activation of MMP-2 was stimulated by the cultivation of MSCs within the collagenous matrix. These results demonstrated that various interactive effects of growth factors and scaffolds influence the cell-biological behaviour of MSCs. It is important to take these complex interactions, which partly differ from differentiated cells, into account in further tissue-engineering approaches.
Heckmann
Leslie
L
Division for Biochemistry of Joint and Connective Tissue Diseases, Department of Orthopedics, University of Ulm, Ulm, Federal Republic of Germany.
Fiedler
Jörg
J
Mattes
Thomas
T
Dauner
Martin
M
Brenner
Rolf E
RE
D000328:Adult; D000077214:Becaplermin; D019485:Bone Morphogenetic Proteins / Q000494:pharmacology; D049109:Cell Proliferation; D003094:Collagen / Q000235:genetics; D024042:Collagen Type I; D005260:Female; D005782:Gels; D055431:Growth Differentiation Factor 6; D006801:Humans; D016023:Integrins / Q000235:genetics; D019344:Lactic Acid; D008297:Male; D053505:Matrix Metalloproteinases, Secreted / Q000235:genetics / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D039902:Multipotent Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology*; D011091:Polyesters; D011108:Polymers; D020574:Proto-Oncogene Proteins c-sis; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology; D019063:Tenascin / Q000235:genetics; D023822:Tissue Engineering; D054457:Tissue Scaffolds; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology*
D019485:Bone Morphogenetic Proteins; D024042:Collagen Type I; C525906:GDF6 protein, human; D005782:Gels; D055431:Growth Differentiation Factor 6; D016023:Integrins; D010982:Platelet-Derived Growth Factor; D011091:Polyesters; D011108:Polymers; D020574:Proto-Oncogene Proteins c-sis; D012333:RNA, Messenger; D011994:Recombinant Proteins; D019063:Tenascin; D053773:Transforming Growth Factor beta1; D000077214:Becaplermin; D019344:Lactic Acid; C033616:poly(lactide); D003094:Collagen; D053505:Matrix Metalloproteinases, Secreted