Meniscal regeneration using tissue engineering with a scaffold derived from a rat meniscus and mesenchymal stromal cells derived from rat bone marrow.
The purpose of this study was to regenerate a meniscus using a scaffold from a normal meniscus and mesenchymal stromal cells derived from bone marrow (BM-MSCs). Thirty Sprague-Dawley rat menisci were excised and freeze-thawed three times with liquid nitrogen to kill the original meniscal cells. Bone marrow was aspirated from enhanced green fluorescent protein transgenic Sprague-Dawley rats. BM-MSCs were isolated, cultured for 2 weeks, and 2 x 10(5) cells were then seeded onto the meniscal scaffolds. Using a fluorescent microscope and immunohistochemical staining, repopulation of enhanced green fluorescent protein positive cells was observed in the superficial zone of the scaffold after 1 week of culture, and then in the deep zone after 2 weeks. At 4 weeks, expression of extracellular matrices was detected histologically and expression of mRNA for aggrecan and type X collagen was detected. Stiffness of the cultured tissue, assessed by the indentation stiffness test, had increased significantly after 2 weeks in culture, and approximated the stiffness of a normal meniscus. From this study, we conclude that a scaffold derived from a normal meniscus seeded with BM-MSCs can form a meniscus approximating a normal meniscus.
Yamasaki
Takuma
T
Department of Orthopaedic Surgery, Graduate School of Biomedical Sciences, Hiroshima University, 1-2-3, Kasumi, Minami-ku, Hiroshima, 734-8551, Japan. tacyama@hiroshima-u.ac.jp
Deie
Masataka
M
Shinomiya
Rikuo
R
Izuta
Yasunori
Y
Yasunaga
Yuji
Y
Yanada
Shinobu
S
Sharman
Patrick
P
Ochi
Mitsuo
M
D000818:Animals; D030801:Animals, Genetically Modified; D001672:Biocompatible Materials / Q000737:chemistry*; D001703:Biophysics / Q000379:methods; D001854:Bone Marrow Cells / Q000166:cytology*; D002356:Cartilage / Q000378:metabolism; D024121:Collagen Type X / Q000096:biosynthesis; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D008856:Microscopy, Fluorescence; D009584:Nitrogen / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012038:Regeneration; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017154:Stromal Cells / Q000166:cytology*; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods*
D001672:Biocompatible Materials; D024121:Collagen Type X; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; D009584:Nitrogen