Chondrogenesis using mesenchymal stem cells and PCL scaffolds.
We tested the in vitro feasibility of porous PCL (poly(epsilon-caprolactone)) as a scaffold for cartilage tissue engineering from mesenchymal stem cells (MSCs) and determined the effects of various surface treatments. Three porous PCL scaffold modifications were examined: (1) PCL/Pluronic F127, (2) PCL/collagen, and (3) PCL/Pluronic F127/collagen, in addition to (4) PCL-only. MSCs (5 x 10(5)) were seeded in PCL scaffolds of pore size 100-150 microm, and after 3 weeks of in vitro culture, MSC-scaffolds were investigated for gross appearance, DNA amount, glycosaminoglycan (GAG) content, chondrogenic gene expression, and histology. Grossly, the cell-scaffold complexes became harder, and were more easily manipulated with a forceps after 3 weeks of culture. The three surface-treated scaffolds had higher DNA contents than did the PCL-only scaffold, and the GAG contents in PCL/collagen and PCL/F127/collagen scaffolds were higher than those seen in the PCL-only scaffold. Real-time PCR showed that Sox-9 and COL2A1 mRNA levels were remarkably elevated in PCL/collagen and PCL/F127/collagen scaffolds versus the PCL-only scaffold. On the other hand, Col1A1 and Col10A1 mRNA levels were lower in the three modified PCL scaffolds than in the PCL-only scaffold. Histological findings generally concurred with GAG and RT-PCR findings, and demonstrated the affinity of PCL-based scaffolds for MSCs and the potentials of these scaffold to induce chondrogenic differentiation. Cells showed more differentiated appearance and more abundant extracellular matrix formation in PCL/collagen and PCL/collagen/F127 scaffolds. Our findings suggest that PCL-based porous scaffolds may be useful carriers for MSC transplantation in the cartilage tissue engineering field, and that collagen-based surface modifications further enhance the chondrogenic differentiation of MSCs.
Kim
Hye-Joung
HJ
Department of Orthopaedics, Dongguk University International Hospital, 814 Siksa-dong, Goyang 410-773, Korea.
Lee
Jin-Ho
JH
Im
Gun-Il
GI
D000368:Aged; D000369:Aged, 80 and over; D001672:Biocompatible Materials / Q000737:chemistry*; D001854:Bone Marrow Cells / Q000502:physiology; D002469:Cell Separation; D002478:Cells, Cultured; D020219:Chondrogenesis / Q000502:physiology*; D024042:Collagen Type I / Q000096:biosynthesis; D004396:Coloring Agents; D004247:DNA / Q000096:biosynthesis / Q000235:genetics; D005260:Female; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000096:biosynthesis / Q000235:genetics; D006025:Glycosaminoglycans / Q000737:chemistry; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D008875:Middle Aged; D010619:Phenazines; D020442:Poloxamer; D011091:Polyesters / Q000737:chemistry*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055758:SOX9 Transcription Factor / Q000096:biosynthesis / Q000235:genetics; D013501:Surface-Active Agents; D054457:Tissue Scaffolds / Q000737:chemistry*
D001672:Biocompatible Materials; D024042:Collagen Type I; D004396:Coloring Agents; D006025:Glycosaminoglycans; D010619:Phenazines; D011091:Polyesters; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D013501:Surface-Active Agents; D020442:Poloxamer; C016240:polycaprolactone; D004247:DNA; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; C009195:safranine T