Selective inhibition of type X collagen expression in human mesenchymal stem cell differentiation on polymer substrates surface-modified by glow discharge plasma.
Recent evidence indicates that a major drawback of current cartilage- and disc-tissue engineering is that human mesenchymal stem cells (MSCs) rapidly express type X collagen-a marker of chondrocyte hypertrophy associated with endochondral ossification. Some studies have attempted to use growth factors to inhibit type X collagen expression, but none to date has addressed the possible effect of the substratum on chondrocyte hypertrophy. Here, we sought to examine the growth and differentiation potential of human MSCs cultured on two polymer types, polypropylene and nylon-6, both of which have been surface-modified by glow discharge plasma treatment in ammonia gas. Cultures were performed for up to 14 days in Dulbecco's modified Eagle medium + 10% fetal bovine serum. Commercial polystyrene culture dishes were used as control. Reverse transcriptase-polymerase chain reaction was used to assess the expression of types I, II, and X collagens and aggrecan using gene-specific primers. Glyceraldehyde-3-phosphate dehydrogenase was used as a housekeeping gene. Types I and X collagens, as well as aggrecan, were found to be constitutively expressed by human MSCs on polystyrene culture dishes. Whereas both untreated and treated nylon-6 partially inhibited type X collagen expression, treated polypropylene almost completely inhibited its expression. These results indicate that plasma-treated polypropylene or nylon-6 may be a suitable surface for inducing MSCs to a disc-like phenotype for tissue engineering of intervertebral discs in which hypertrophy is suppressed.
Nelea
Valentin
V
Department of Engineering Physics, Ecole Polytechnique, 2900 Edouard-Montpetit, Montréal, Québec H3T 1J4, Canada.
Luo
Li
L
Demers
Caroline N
CN
Antoniou
John
J
Petit
Alain
A
Lerouge
Sophie
S
R Wertheimer
Michael
M
Mwale
Fackson
F
D000368:Aged; D053674:Aggrecans; D001842:Bone and Bones / Q000378:metabolism; D002209:Caprolactam / Q000031:analogs & derivatives / Q000737:chemistry; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D048429:Cell Size; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism; D024121:Collagen Type X / Q000037:antagonists & inhibitors* / Q000737:chemistry; D003470:Culture Media / Q000737:chemistry; D016326:Extracellular Matrix Proteins / Q000737:chemistry; D006801:Humans; D006984:Hypertrophy; D037181:Lectins, C-Type; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D008858:Microscopy, Phase-Contrast; D008875:Middle Aged; D008956:Models, Chemical; D010641:Phenotype; D011108:Polymers / Q000737:chemistry*; D011126:Polypropylenes / Q000737:chemistry; D011509:Proteoglycans / Q000737:chemistry; D012313:RNA / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology; D013499:Surface Properties; D013997:Time Factors; D023822:Tissue Engineering
D053674:Aggrecans; D024121:Collagen Type X; D003470:Culture Media; D016326:Extracellular Matrix Proteins; D037181:Lectins, C-Type; D011108:Polymers; D011126:Polypropylenes; D011509:Proteoglycans; C009916:nylon 6; D012313:RNA; D002209:Caprolactam