Chondrogenic differentiation on perlecan domain I, collagen II, and bone morphogenetic protein-2-based matrices.
Extracellular matrix (ECM) molecules in cartilage cooperate with growth factors to regulate chondrogenic differentiation and cartilage development. Domain I of perlecan (Pln) bears heparan sulfate chains that bind and release heparin binding growth factors (HBGFs). We hypothesized that Pln domain I (PlnDI) might be complexed with collagen II (P-C) fibrils to improve binding of bone morphogenetic protein-2 (BMP-2) and better support chondrogenesis and cartilage-like tissue formation in vitro. Our results showed that P-C fibrils bound more BMP-2 than collagen II fibrils alone, and better sustained BMP-2 release. Polylactic acid (PLA)-based scaffolds coated with P-C fibrils immobilized more BMP-2 than either PLA scaffolds or PLA scaffolds coated with collagen II fibrils alone. Multipotential mouse embryonic mesenchymal cells, C3H10T1/2, were cultured on 2-dimensional P-C fibrils or 3-dimensional P-C/BMP-2-coated (P-C-B) PLA scaffolds. Chondrogenic differentiation was indexed by glycosaminoglycan (GAG) production, and expression of the pro-chondrogenic transcription factor, Sox9, as well as cartilaginous ECM proteins, collagen II, and aggrecan. Immunostaining for aggrecan, perlecan, tenascin, and collagen X revealed that both C3H10T1/2 cells and primary mouse embryonic fibroblasts cultured on P-C-B fibrils showed the highest expression of chondrogenic markers among all treatment groups. Safranin O-Fast Green staining indicated that cartilage-like tissue was formed in the P-C-B scaffolds, while no obvious cartilage-like tissue formed in other scaffolds. We conclude that P-C fibrils provide an improved biomimetic material for the binding and retention of BMP-2 and support chondrogenic differentiation.
Yang
Weidong
W
Department of Biological Sciences, University of Delaware, Newark, Delaware 19716, USA.
Gomes
Ronald R
RR
Brown
Anissa J
AJ
Burdett
Ashley R
AR
Alicknavitch
Michael
M
Farach-Carson
Mary C
MC
Carson
Daniel D
DD
D000818:Animals; D000943:Antigens, Differentiation / Q000378:metabolism; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D002460:Cell Line; D020219:Chondrogenesis / Q000502:physiology*; D024043:Collagen Type II; D004622:Embryo, Mammalian / Q000166:cytology / Q000502:physiology*; D005109:Extracellular Matrix; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology*; D019812:Heparan Sulfate Proteoglycans; D051379:Mice; D013234:Stem Cells / Q000166:cytology / Q000502:physiology*; D016212:Transforming Growth Factor beta
D000943:Antigens, Differentiation; C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D024043:Collagen Type II; D019812:Heparan Sulfate Proteoglycans; D016212:Transforming Growth Factor beta; C071927:perlecan