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475 THE EFFECT OF NAOH ON PROLIFERATION AND DIFFERENTIATION OF MC3T3-E1 PREOSTEOBLASTIC CELLS GROWN ON THREE-DIMENSIONAL POLY(L-LACTIDE-CO-GLYCOLIDE) SCAFFOLDS.
  1. M. Hakimi1,
  2. S. Farnad1,
  3. K. Caprizo1,2,
  4. W. Huang1,2,
  5. C. Huang1,2,
  6. K. Ishida2,
  7. T. A. Miller1,2,
  8. G. H. Rudkin1,2
  1. 1David Geffen School of Medicine at UCLA, Los Angeles, CA
  2. 2Plastic Surgery Section, VA Greater LA Health System, Los Angeles, CA

Abstract

Background In creating a biosynthetic scaffold that promotes osteogenesis surface topography is an important variable. It has been demonstrated that chemical etching of poly(l-lactide-co-glycolide) (PLGA) with NaOH enhances adhesion and function of numerous cells types in two-dimensional systems (hepatocytes, endothelial cells, vascular and bladder smooth muscle cells) and three-dimensional (3-D) systems (chondrocytes). This study compares the biological behavior of osteoblasts grown on NaOH-treated PLGA scaffolds and conventional PLGA scaffolds by measuring osteoblast adhesion, differentiation, and proliferation of MC3T3-E1 cells cultured on NaOH-treated 3D PLGA scaffolds and untreated scaffolds.

Methods MC3T3-E1 mouse preosteoblast cells were seeded onto 3-D PLGA scaffolds ± 1 M NaOH for 10 minutes. Quantitative real-time RT PCR was performed to measure mRNA expression of osteogenic marker genes. In addition, cell numbers were determined at varying time points.

Results Pretreatment of 3D PLGA scaffolds with NaOH resulted in statistically significant up-regulation of mRNA expression of alkaline phosphatase (ALP), bone sialoprotein (BSP), osteocalcin (OCN), and vascular endothelial growth factor (VEGF). A striking increase in mineralized cell matrix deposition in the NaOH group was also observed. Cell number was statistically higher in the NaOH group immediately after cell seeding, suggesting improved adhesion. During the first 24 hours of culture, cell proliferative rates were higher in the control group than the NaOH group.

Conclusion Chemical etching of PLGA scaffolds with NaOH enhances adhesion and differentiation and slows proliferation of preosteoblast cells. NaOH treatment may represent an inexpensive way to improve scaffolds for use in bone tissue engineering.

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