Pretreatment of poly(l-lactide-co-glycolide) scaffolds with sodium hydroxide enhances osteoblastic differentiation and slows proliferation of mouse preosteoblast cells.
BACKGROUND
Surface topography is important in the creation of a scaffold for tissue engineering. Chemical etching of poly(l-lactide-co-glycolide) with sodium hydroxide has been shown to enhance adhesion and function of numerous cell types. The authors investigated the effects of sodium hydroxide pretreatment of three-dimensional poly(l-lactide-co-glycolide) scaffolds on the adhesion, differentiation, and proliferation of MC3T3-E1 murine preosteoblasts.


METHODS
MC3T3-E1 cells were seeded onto three-dimensional poly(l-lactide-co-glycolide) scaffolds with and without 1 M sodium hydroxide pretreatment. Cells were then cultured in osteogenic medium and harvested at varying time points for RNA extraction. Quantitative real-time reverse-transcriptase polymerase chain reaction was performed to measure mRNA expression of several osteogenic marker genes. In addition, cell numbers were determined at varying time points during the culture period. All experiments were performed in triplicate.


RESULTS
Pretreatment of three-dimensional poly(l-lactide-co-glycolide) scaffolds with sodium hydroxide resulted in statistically significant up-regulation of mRNA expression of alkaline phosphatase, bone sialoprotein, osteocalcin, and vascular endothelial growth factor during the first 10 days of culture. Histologic analysis demonstrated a striking increase in mineralized cell matrix deposition in the sodium hydroxide-treated group. Cell number was statistically higher in the sodium hydroxide-treated group immediately after cell seeding, suggesting improved adhesion. During the first 24 hours of culture, cells grew faster in the control group than in the sodium hydroxide-treated group.


CONCLUSIONS
Chemical etching of poly(l-lactide-co-glycolide) scaffolds with sodium hydroxide strongly influences the behavior of MC3T3-E1 preosteoblasts in vitro by enhancing adhesion and differentiation and slowing proliferation. Sodium hydroxide treatment may represent a simple and inexpensive way of improving scaffolds for use in bone tissue engineering.
Carpizo
Katherine H
KH
Los Angeles, Calif. From the Plastic Surgery Section, Veterans Affairs Greater Los Angeles Healthcare System, and the Division of Plastic and Reconstructive Surgery, Department of Surgery, University of California, Los Angeles.
Saran
Madeleine J
MJ
Huang
Weibiao
W
Ishida
Kenji
K
Roostaeian
Jason
J
Bischoff
David
D
Huang
Catherine K
CK
Rudkin
George H
GH
Yamaguchi
Dean T
DT
Miller
Timothy A
TA
D000818:Animals; D001672:Biocompatible Materials / Q000494:pharmacology; D002424:Caustics / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D049109:Cell Proliferation; D004337:Drug Carriers; D015870:Gene Expression / Q000187:drug effects; D058574:Integrin-Binding Sialoprotein; D019344:Lactic Acid / Q000494:pharmacology*; D051379:Mice; D010006:Osteoblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects / Q000235:genetics*; D011100:Polyglycolic Acid / Q000494:pharmacology*; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011108:Polymers / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012795:Sialoglycoproteins / Q000096:biosynthesis / Q000235:genetics; D012972:Sodium Hydroxide / Q000494:pharmacology*; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects; D023822:Tissue Engineering; D054457:Tissue Scaffolds
D001672:Biocompatible Materials; D002424:Caustics; D004337:Drug Carriers; C546872:Ibsp protein, mouse; D058574:Integrin-Binding Sialoprotein; D011108:Polymers; D012333:RNA, Messenger; D012795:Sialoglycoproteins; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D012972:Sodium Hydroxide