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Objective: To investigate the effects of porous poly lactide-co-glycolide (PLGA) modified by type I collagen on the adhesion, proliferation, and differentiation of rabbit marrow-derived mesenchymal stem cells (MSCs). Methods: The third generation MSCs isolated from mature rabbits by density gradient centrifugation were cultured at different initial concentrations on (0.3 cm)×(1.2 cm)×(2.0 cm) 3-D porous PLGA coated by type I collagen in RPMI 1640 containing 10% fetal calf serum, while cultured on PLGA without type I collagen as control. The cells adhesive and proliferative behavior at 7, 14, and 21 days after inoculation was assessed by determining the incorporation rate of [~3H]-TdR. In order to examine MSCs differentiation, the expression of osteoblasts marker genes, osteocalcin (OCN), alkaline phosphatase (ALP), osteopontin (OPN) mRNA, were evaluated by reverse transcription-polymerase chain reaction (RT-PCR), and further more, the cell morphology at 21 days was also observed by scanning electron microscope (SEM). Results: Type I collagen promoted cell adhesion on PLGA. The valve was significantly higher than controls (6 h, 2144 cpm±141cpm vs. 1797 cpm±118 cpm, P=(0.017); 8 h, 2311 cpm±(113 cpm) vs. 1891 cpm±103 cpm, P=(0.01)). The cells which cultured on PLGA coated with type I collagen showed significantly higher cell proliferation than controls on the 7th day ((1021 cpm)±159 cpm vs. 451 cpm±67 cpm, P=(0.002)), the 14th day (1472 cpm±(82 cpm) vs. 583 cpm±67 cpm, P<(0.001)) and 21th day (1728 cpm±78 cpm vs. 632 cpm±55 cpm, P<(0.001)). Osteoblasts markers, OCN, ALP, OPN mRNA, were all detected on PLGA coated by type I collagen on the 21th day, but OCN, OPN mRNA could not be found in controls. Spindle and polygonal cells well distributed on the polymer coated by type I collagen while cylindric or round cells in controls. Conclusions: Type I collagen is effective in promoting the adhesion, proliferation and differentiation of MSCs on PLGA.
Objective: To investigate the effects of porous polylactide-co-glycolide (PLGA) modified by type I collagen on the adhesion, proliferation, and differentiation of rabbit marrow-derived mesenchymal stem cells (MSCs). Methods: The third generation MSCs isolated from mature rabbits by density gradient centrifugation were cultured at different initial concentrations on (0.3 cm) × (1.2 cm) × (2.0 cm) 3-D porous PLGA coated by type I collagen in RPMI 1640 containing 10% fetal calf serum, while cultured on PLGA without type I collagen as control. The cells adhesive and proliferative behavior at 7, 14, and 21 days after inoculation was assessed by incorporation rate of [~ 3H] -TdR. In order to examine MSCs differentiation, the expression of osteoblasts marker genes, osteocalcin (OCN), alkaline phosphatase (ALP), osteopontin (OPN) mRNA, were evaluated by reverse transcription-polymerase chain reaction (RT- PCR), and further more, the cell morphology at 21 days was also observed by The valve was significantly higher than controls (6 h, 2144 cpm ± 141 cpm vs. 1797 cpm ± 118 cpm, P = (0.017); 8 h, 2311 cpm ± 113 cpm vs. 1891 cpm ± 103 cpm P = 0.01). The cells which cultured on PLGA coated with type I collagen showed significantly higher cell proliferation than controls on the 7th day ((1021 cpm) ± (1472 cpm ± (82 cpm) vs. 583 cpm ± 67 cpm, P <(0.001)) and 21 th day (1728 cpm ± 78 cpm vs. 632 cpm ± 55 cpm, P <(0.001)). Osteoblasts markers, OCN, ALP, OPN mRNA, were all detected on PLGA coated by type I collagen on the 21st day, but OCN, OPN mRNA could not be found in controls. Spindle and polygonal cells well distributed on the polymer coated by type I collagen while cylindric or round cells in controls. Conclusions: Type I collagen is effective in promoting the adhesion, proliferation and differentiation of MSCs on PLGA.