Cytotoxicity testing with three-dimensional cultures of transfected pulp-derived cells.
SV40 large T-antigen-transfected bovine pulp-derived cells were grown three-dimensionally on polyamide meshes. For optimal cell growth, various cell numbers and mesh coatings were tested. Next the three-dimensional cultures were used in a dentin barrier test device, and the system was evaluated by testing a set of dental filling materials. After 24 hr exposure with or without perfusion of the pulpal part of the test device, cell survival was evaluated using an MTT assay. In all experiments pulp-derived cells transfected with SV40 large T-antigen grew three-dimensionally on polyamide meshes and showed growth kinetics similar to those on cell culture plates with lag, log, and plateau phases (reached after about 14 days of incubation). Cross-sections of the three-dimensional cell cultures revealed about 15 to 20 cell layers. In vitro cytotoxicity tests resulted in cell survival rates which are in good agreement with in vivo data and with results obtained from cytotoxicity tests with three-dimensional cultures of human foreskin fibroblasts.
Schuster
U
U
Department of Operative Dentistry and Periodontology, University of Regensburg, Germany.
Schmalz
G
G
Thonemann
B
B
Mendel
N
N
Metzl
C
C
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics; D002417:Cattle; D002452:Cell Count; D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004396:Coloring Agents; D003470:Culture Media; D003782:Dental Pulp / Q000166:cytology / Q000187:drug effects*; D003804:Dentin; D015194:Diffusion Chambers, Culture; D005347:Fibroblasts / Q000187:drug effects; D006801:Humans; D010477:Perfusion; D019800:Phenol / Q000633:toxicity; D012387:Root Canal Filling Materials / Q000633:toxicity*; D012680:Sensitivity and Specificity; D012867:Skin / Q000166:cytology / Q000187:drug effects; D018709:Statistics, Nonparametric; D013778:Tetrazolium Salts; D013844:Thiazoles; D013997:Time Factors; D014048:Tolonium Chloride; D014162:Transfection; D015035:Zinc Oxide-Eugenol Cement / Q000633:toxicity; D015036:Zinc Phosphate Cement / Q000633:toxicity
D000952:Antigens, Polyomavirus Transforming; D004396:Coloring Agents; D003470:Culture Media; D012387:Root Canal Filling Materials; D013778:Tetrazolium Salts; D013844:Thiazoles; D015035:Zinc Oxide-Eugenol Cement; D014048:Tolonium Chloride; D019800:Phenol; D015036:Zinc Phosphate Cement; C022616:thiazolyl blue