Cryopreservation of periodontal ligament cells with magnetic field for tooth banking.
The purpose of this study was to establish a long-term tooth cryopreservation method that can be used for tooth autotransplantation. Human periodontal ligament (PDL) cells were frozen in 10% dimethyl sulfoxide (Me(2)SO) using a programmed freezer with a magnetic field. Cells were cryopreserved for 7 days at -150 degrees C. Immediately after thawing, the number of surviving cells was counted and the cells were cultured; cultured cells were examined after 48 h. Results indicated that a 0.01 mT of a magnetic field, a 15-min hold-time, and a plunging temperature of -30 degrees C led to the greatest survival rate of PDL cells. Based on these findings, whole teeth were cryopreserved under the same conditions for 1 year. The organ culture revealed that the PDL cells of cryopreserved tooth with a magnetic field could proliferate as much as a fresh tooth, although the cells did not appear in the cryopreserved tooth without a magnetic field. Histological examination and the transmission electron microscopic image of cryopreserved tooth with a magnetic field did not show any destruction of cryopreserved cells. In contrast, severe cell damage was seen in cells frozen without a magnetic field. These results indicated that a magnetic field programmed freezer is available for tooth cryopreservation.
Kaku
M
M
Department of Orthodontics and Craniofacial Developmental Biology, Hiroshima University Graduate School of Biomedical Science, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, Japan. mkaku@hiroshima-u.ac.jp
Kamada
H
H
Kawata
T
T
Koseki
H
H
Abedini
S
S
Kojima
S
S
Motokawa
M
M
Fujita
T
T
Ohtani
J
J
Tsuka
N
N
Matsuda
Y
Y
Sunagawa
H
H
Hernandes
R A M
RA
Ohwada
N
N
Tanne
K
K
D002470:Cell Survival / Q000187:drug effects / Q000528:radiation effects; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004574:Electromagnetic Fields; D006801:Humans; D008280:Magnetics; D046529:Microscopy, Electron, Transmission; D009924:Organ Culture Techniques; D010513:Periodontal Ligament / Q000166:cytology* / Q000187:drug effects / Q000528:radiation effects; D014015:Tissue Banks
D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide