Comparison of chondrogensis in static and perfused bioreactor culture

被引:195
作者
Pazzano, D
Mercier, KA
Moran, JM
Fong, SS
DiBiasio, DD
Rulfs, JX
Kohles, SS
Bonassar, LJ
机构
[1] Univ Massachusetts, Sch Med, Ctr Tissue Engn, Worcester, MA 01655 USA
[2] Worcester Polytech Inst, Dept Chem Engn, Worcester, MA 01608 USA
[3] Worcester Polytech Inst, Dept Biol, Worcester, MA 01608 USA
[4] Worcester Polytech Inst, Dept Biomed Engn, Worcester, MA 01608 USA
关键词
D O I
10.1021/bp000082v
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
As a result of the low yield of cartilage from primary patient harvests and a high demand for autologous cartilage for reconstructive surgery and structural repair, primary explant cartilage must be augmented by tissue engineering techniques. In this study, chondrocytes seeded on PLLA/PGA scaffolds in static culture and a direct perfusion bioreactor were biochemically and histologically analyzed to determine the effects of fluid flow and media pH on matrix assembly. A gradual media pH change was maintained in the bioreactor within 7.4-6.96 over 2 weeks compared to a more rapid decrease from 7.4 to 6.58 in static culture over 3 days. Seeded scaffolds subjected to 1 mu m/s flow demonstrated a 118% increase (p < 0.05) in DNA content, a 184% increase (p < 0.05) in GAG content, and a 155% (p < 0.05) increase in hydroxyproline content compared to static culture. Distinct differences were noted in tissue morphology, including more intense staining for proteoglycans by safranin-O and alignment of cells in the direction of media flow. Culture of chondrocyte seeded matrices thus offers the possibility of rapid in vitro expansion of donor cartilage for the repair of structural defects, tracheal injury, and vascularized tissue damage.
引用
收藏
页码:893 / 896
页数:4
相关论文
共 13 条
[1]  
BLOEBAUM RD, 1987, STAIN TECH, V62, P401
[2]   Tissue-engineered nipple reconstruction [J].
Cao, YL ;
Lach, E ;
Kim, TH ;
Rodríguez, A ;
Arévalo, CA ;
Vacanti, CA .
PLASTIC AND RECONSTRUCTIVE SURGERY, 1998, 102 (07) :2293-2298
[3]   Transplantation of chondrocytes utilizing a polymer-cell construct to produce tissue-engineered cartilage in the shape of a human ear [J].
Cao, YL ;
Vacanti, JP ;
Paige, KT ;
Upton, J ;
Vacanti, CA .
PLASTIC AND RECONSTRUCTIVE SURGERY, 1997, 100 (02) :297-302
[4]   IMPROVED QUANTITATION AND DISCRIMINATION OF SULFATED GLYCOSAMINOGLYCANS BY USE OF DIMETHYLMETHYLENE BLUE [J].
FARNDALE, RW ;
BUTTLE, DJ ;
BARRETT, AJ .
BIOCHIMICA ET BIOPHYSICA ACTA, 1986, 883 (02) :173-177
[5]  
Freed LE, 1997, IN VITRO CELL DEV-AN, V33, P381
[6]   NEOCARTILAGE FORMATION INVITRO AND INVIVO USING CELLS CULTURED ON SYNTHETIC BIODEGRADABLE POLYMERS [J].
FREED, LE ;
MARQUIS, JC ;
NOHRIA, A ;
EMMANUAL, J ;
MIKOS, AG ;
LANGER, R .
JOURNAL OF BIOMEDICAL MATERIALS RESEARCH, 1993, 27 (01) :11-23
[7]   MECHANICAL AND PHYSICOCHEMICAL DETERMINANTS OF THE CHONDROCYTE BIOSYNTHETIC RESPONSE [J].
GRAY, ML ;
PIZZANELLI, AM ;
GRODZINSKY, AJ ;
LEE, RC .
JOURNAL OF ORTHOPAEDIC RESEARCH, 1988, 6 (06) :777-792
[8]   THE ROLE OF CARTILAGE STREAMING POTENTIAL, FLUID-FLOW AND PRESSURE IN THE STIMULATION OF CHONDROCYTE BIOSYNTHESIS DURING DYNAMIC COMPRESSION [J].
KIM, YJ ;
BONASSAR, LJ ;
GRODZINSKY, AJ .
JOURNAL OF BIOMECHANICS, 1995, 28 (09) :1055-1066
[9]   FLUOROMETRIC ASSAY OF DNA IN CARTILAGE EXPLANTS USING HOECHST-33258 [J].
KIM, YJ ;
SAH, RLY ;
DOONG, JYH ;
GRODZINSKY, AJ .
ANALYTICAL BIOCHEMISTRY, 1988, 174 (01) :168-176
[10]  
Klagsbrun M, 1979, Methods Enzymol, V58, P560