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168 DIFFERENTIATION OF MC3T3-E1 PREOSTEOBLASTIC CELLS ON TWO-DIMENSIONAL POLY(LACTIDE-CO-GLYCOLIDE) (PLGA) FILMS AND THREE-DIMENSIONAL PLGA SCAFFOLDS: A REAL-TIME REVERSE TRANSCRIPTASE-POLYMERASE CHAIN REACTION STUDY.
  1. S. A. Farnad1,2,
  2. M. Hakimi1,2,
  3. W. Huang2,3,
  4. G. H. Rudkin2,3,
  5. K. Ishida2,
  6. C. Huang2,3,
  7. D. T. Yamaguchi2,
  8. T. A. Miller2,3
  1. 1David Geffen School of Medicine at UCLA, Los Angeles, CA
  2. 2 Plastic Surgery Research Laboratory, VA Greater LA Healthcare System, Los Angeles, CA
  3. 3Division of Plastic Surgery, Department of Surgery, UCLA, Los Angeles, CA.

Abstract

Purpose This study aimed to investigate the expression of a number of osteogenic and angiogenic markers in MC3T3-E1 cells cultured on two-dimensional (2-D) (PLGA films and three-dimensional (3-D) PLGA scaffolds.

Methods MC3T3-E1 cells were incubated in mineralized medium in the absence or presence of 100 ng/mL recombinant human BMP-2 for 2 weeks. Cells were cultured either on 2-D PLGA films or in 3-D PLGA scaffolds and harvested on days 0, 1, 2, 3, 5, 7, 10, and 14. Total RNA was extracted and subjected to quantitative real-time reverse transcriptase-polymerase chain reaction analysis for expression of genes involved in osteogenesis and angiogenesis.

Results Expression of BSP and OCN increased when cells cultured on 2-D PLGA films were subjected to differentiation medium containing ascorbic acid and beta-glycerol phosphate for a period of 2 weeks. Treatment with recombinant human BMP-2 resulted in further increases in expression of both genes after day 5. Cells cultured on 3-D PLGA scaffolds showed significantly slowed differentiation as measured by expression of the above two genes. Both genes remained highly responsive to BMP-2 treatment after day 5. OPN expression was elevated when cells were transplanted into 3-D scaffolds. Its expression in cells cultured in 3-D scaffolds was less responsive to BMP-2 treatment than on 2-D films. Expression of VEGF was increased as differentiation proceeded on 2-D films and was enhanced with BMP-2 treatment. Its expression was significantly induced when cells were cultured in 3-D scaffolds. A 2-week culture of cells in 3-D scaffolds resulted in decreased VEGF expression. BMP-2 inhibited VEGF expression in 3-D scaffolds at days 10 and 14.

Conclusions The results indicate that MC3T3-E1 cells commit to osteogenic differentiation at a slower rate in 3-D scaffolds than in 2-D scaffolds.

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