In vitro analysis of an allogenic scaffold for tissue-engineered meniscus replacement.
Scaffolds play a key role in the field of tissue engineering. Particularly for meniscus replacement, optimal scaffold properties are critical. The aim of our study was to develop a novel scaffold for replacement of meniscal tissue by means of tissue engineering. Emphasis was put on biomechanical properties comparable to native meniscus, nonimmunogenecity, and the possibility of seeding cells into and cultivating them within the scaffold (nontoxicity). For this purpose, native ovine menisci were treated in vitro in a self-developed enzymatic process. Complete cell removal was achieved and shown both histologically and electron microscopically (n = 15). Immunohistochemical reaction (MHC 1/MHC 2) was positive for native ovine meniscus and negative for the scaffold. Compared to native meniscus (25.8 N/mm) stiffness of the scaffold was significantly increased (30.2 N/mm, p < 0.05, n = 10). We determined the compression (%) of the native meniscus and the scaffold under a load of 7 N. The compression was 23% for native meniscus and 29% for the scaffold (p < 0.05, n = 10). Residual force of the scaffold was significantly lower (5.2 N vs. 4.9 N, p < 0.05, n = 10). Autologous fibrochondrocytes were needle injected and successfully cultivated within the scaffolds over a period of 4 weeks (n = 10). To our knowledge, this study is the first to remove cells and immunogenetic proteins (MHC 1/MHC 2) completely out of native meniscus and preserve important biomechanical properties. Also, injected cells could be successfully cultivated within the scaffold. Further in vitro and in vivo animal studies are necessary to establish optimal cell sources, sterilization, and seeding techniques. Cell differentiation, matrix production, in vivo remodeling of the construct, and possible immunological reactions after implantation are subject of further studies.
Department of Orthopaedic Sports Medicine, Technical University of Munich, Munich, Germany. dirk.maier@uniklinik-freiburg.de
Dirk
D
Maier
Klaus
K
Braeun
Erwin
E
Steinhauser
Peter
P
Ueblacker
Michael
M
Oberst
Peter C
PC
Kreuz
Nadine
N
Roos
Vladimir
V
Martinek
Andreas B
AB
Imhoff
D000818:Animals; D001696:Biomechanical Phenomena; D006025:Glycosaminoglycans / Q000378:metabolism; D007150:Immunohistochemistry; D008422:Materials Testing; D008592:Menisci, Tibial / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D008855:Microscopy, Electron, Scanning; D012756:Sheep; D023822:Tissue Engineering; D054457:Tissue Scaffolds; D014182:Transplantation, Autologous
D006025:Glycosaminoglycans