Physicobiochemical synergism through gene therapy and functional tissue engineering for in vitro chondrogenesis.
Mechanical and chemical stimulation have been shown to enhance in vitro chondrogenesis. The aim of this study was to analyze and compare combined physicobiochemical effects. Bovine articular chondrocytes were retrovirally transduced to express bone morphogenetic protein-2 (BMP-2) or left as naïve controls. Cells were seeded in three-dimensional polyurethane scaffolds and further cultured under static conditions or exposed to dynamic compression and shear in a joint-specific bioreactor. Four groups: control (A), load (B), BMP-2-infected (C), and BMP-2-infected plus load (D) were analyzed for DNA and glycosaminoglycan (GAG) content; collagen I, II, and X; aggrecan, (cartilage oligomeric protein (COMP), superficial zone protein, matrix metalloproteinase (MMP)-3; MMP-13 mRNA; histology; and immunohistochemistry at 7, 21, and 35 days post-seeding. Synergistic effects (D) were higher than the sum of the individual treatments (B and C) for GAG/DNA, collagen II, and COMP. Histology revealed a functional organization in D including an intense safranin O staining in C and D superior to that in A and B. Immunostaining for collagen II and aggrecan was detected in C and D and was strongest in D. The results show that both stimuli augment in vitro chondrogenesis better than in controls. Biochemical manipulation proved to be predominantly more effective than load, and synergistic effects were demonstrated.
Salzmann
Gian M
GM
Department of Orthopaedic and Trauma Surgery, University Medical Center, Albert-Ludwigs University Freiburg, Freiburg, Germany. giansalzmann@yahoo.com
Nuernberger
Benedikt
B
Schmitz
Philipp
P
Anton
Martina
M
Stoddart
Martin J
MJ
Grad
Sibylle
S
Milz
Stefan
S
Tischer
Thomas
T
Vogt
Stephan
S
Gansbacher
Bernd
B
Imhoff
Andreas B
AB
Alini
Mauro
M
D000818:Animals; D002417:Cattle; D019902:Chondrocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D020219:Chondrogenesis / Q000187:drug effects; D016616:Cryoultramicrotomy; D004247:DNA / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D015316:Genetic Therapy; D006025:Glycosaminoglycans / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007150:Immunohistochemistry; D011140:Polyurethanes / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D023822:Tissue Engineering; D054457:Tissue Scaffolds; D014161:Transduction, Genetic
D006025:Glycosaminoglycans; D011140:Polyurethanes; D012333:RNA, Messenger; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D004247:DNA