Effects of FGF-2 and OP-1 in vitro on donor source cartilage for auricular reconstruction tissue engineering.
OBJECTIVE
Microtia is a congenital partial or total loss of the external ear with current treatment approaches involving autologous construction from costal cartilage. Alternatively, tissue engineering provides possible use of normal or microtia auricular chondrocytes harvested from patients. This study investigated effects in vitro of basic fibroblast growth factor (FGF-2) and osteogenic protein 1 (OP-1) on human pediatric normal and microtia auricular chondrocytes and their potential proliferation and differentiation for cellular expansion. A working hypothesis was that FGF-2 promotes proliferation and OP-1 maintains an auricular phenotype of these cells.


METHODS
Two patients, one undergoing otoplasty and one an ear construction, yielded normal and microtia auricular chondrocytes, respectively. The two donor sets of isolated chondrocytes were equally divided into four experimental cell groups. These were controls without added growth factors and cells supplemented with FGF-2, OP-1 or FGF-2/OP-1 combined. Cells were cultured 3, 5, 7, and 10 days (3 replicates/time point), counted and assayed by RT-qPCR to determine elastin and types II and III collagen gene expression.


RESULTS
Compared to control counterparts, normal and microtia chondrocytes with OP-1 alone were similar in numbers and varied in elastin and types II and III collagen expression over all culture times. Compared to respective controls and chondrocyte groups with OP-1 alone, normal and microtia cell groups with FGF-2 had statistically significant (p<0.05) enhanced proliferation and statistically significant (p<0.05) decreased elastin and types II and III collagen expression over 10 days of culture.


CONCLUSIONS
FGF-2 effects on normal and microtia chondrocytes support its use for increasing cell numbers while OP-1 maintains a chondrocyte phenotype, otherwise marked by increasing type III collagen expression and cellular dedifferentiation to fibroblasts in culture.
Department of Integrative Medical Sciences, Northeast Ohio Medical University, Rootstown, OH, United States.
Mark
M
Shasti
Department of Polymer Science, University of Akron, Akron, OH, United States.
Robin
R
Jacquet
Department of Polymer Science, University of Akron, Akron, OH, United States.
Phillip
P
McClellan
Department of Integrative Medical Sciences, Northeast Ohio Medical University, Rootstown, OH, United States.
Julianne
J
Yang
Department of Plastic and Reconstructive Surgery, Kinki University Medical School, Osaka-sayama, Osaka, Japan.
Seika
S
Matsushima
Department of Plastic and Reconstructive Surgery, Kinki University Medical School, Osaka-sayama, Osaka, Japan.
Noritaka
N
Isogai
Division of Plastic and Reconstructive Surgery, Children's Hospital Medical Center, Akron, OH, United States.
Ananth
A
Murthy
Department of Integrative Medical Sciences, Northeast Ohio Medical University, Rootstown, OH, United States; Department of Polymer Science, University of Akron, Akron, OH, United States. Electronic address: wlandis@uakron.edu.
William J
WJ
Landis
D055419:Bone Morphogenetic Protein 7 / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D002648:Child; D019902:Chondrocytes / Q000187:drug effects* / Q000502:physiology; D000013:Congenital Abnormalities / Q000175:diagnosis / Q000601:surgery*; D065817:Congenital Microtia; D004423:Ear / Q000002:abnormalities* / Q000601:surgery; D004425:Ear Cartilage / Q000601:surgery; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D006801:Humans; D008297:Male; D019651:Plastic Surgery Procedures / Q000379:methods*; D012016:Reference Values; D014019:Tissue Donors; D023822:Tissue Engineering / Q000379:methods*
D055419:Bone Morphogenetic Protein 7; D016222:Fibroblast Growth Factor 2
Chondrocytes; FGF-2; Microtia; OP-1; Tissue engineering