PMID- 12929739
OWN - NLM
STAT- MEDLINE
DCOM- 20030904
LR  - 20160804
IS  - 1081-5589 (Print)
IS  - 1081-5589 (Linking)
VI  - 51
IP  - 4
DP  - 2003 Jul
TI  - Effects of hyperbaric oxygen on proliferative and apoptotic activities and 
      reactive oxygen species generation in mouse fibroblast 3T3/J2 cell line.
PG  - 227-32
LID - 10.1136/jim-51-04-24 [doi]
AB  - BACKGROUND: Hyperbaric oxygen (HBO) therapy is widely used to treat problem 
      wounds associated with pathologic conditions compromising blood supply and tissue 
      oxygenation because increased tissue oxygen levels enhance collagen synthesis, 
      cell proliferation, and angiogenesis. However, little is known about the dose of 
      hyperoxia needed to achieve optimal therapeutic effects. Moreover, HBO, by 
      enhancing the production of reactive oxygen species (ROS), may also exert 
      cytotoxic effects. In vitro models are simplified systems that may aid the 
      development of treatment protocols with HBO. Hence, we have investigated the 
      effects of HBO on the growth and ROS production of the 3T3/J2 fibroblast cell 
      line in relation to the pressure and the duration of exposure. METHODS: 3T3/J2 
      fibroblasts were plated (5 x 10(3) cells/cm2) on six-well microtiter plates in 
      phosphate buffered saline (PBS), put in a compression chamber, and exposed to 
      100% oxygen at a pressure of 1.0 or 2.5 atmosphere absolute (ATA) for 15, 30, 60, 
      or 120 minutes. Then the cells were incubated in Dulbecco's modified minimum 
      essential medium (DMEM) for 24, 48, or 72 hours, and at the end of the post-HBO 
      incubation period, their number was determined. In other experiments, cells were 
      detached just after HBO exposure, seeded on 60 mm Petri dishes, and cultured for 
      10 days in DMEM, and the colony forming units were counted. The effects of HBO 
      exposure (2.5 ATA) on the apoptotic rate of cultured cells were investigated by 
      terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and 
      enzyme-linked immunosorbent assays. To measure ROS production, 60 minutes before 
      HBO exposure, 2',7'-dichlorofluorescin (DCF) diacetate (200 nmol/mL) was added to 
      PBS, and after HBO exposure (2.5 ATA), cells were lysated, and 
      fluorescence-emission intensity was measured and converted to micromol DCF/microg 
      protein. RESULTS: At 1.0 ATA, all HBO exposures increased the proliferation rate 
      of cultured fibroblasts and their clonal growth efficiency. At 2.5 ATA, 15-minute 
      exposure to HBO was ineffective, whereas 30- and 60-minute exposures raised the 
      proliferation rate and clonal growth efficiency. Conversely, a 120-minute 
      exposure significantly decreased these parameters compared with control cultures. 
      The exposure of cells to HBO at 2.5 ATA for 120 minutes raised the apoptotic rate 
      of cultured fibroblasts, whereas shorter exposure times were ineffective. All 
      exposure periods to HBO at 2.5 ATA enhanced ROS production from cultured 
      fibroblasts. CONCLUSIONS: Collectively, our findings allow us to conclude that 
      (1) all of the exposure periods to HBO at 1.0 ATA or 30- and 60-minute periods at 
      2.5 ATA enhance cell growth, (2) 120-minute exposure to HBO at 2.5 ATA exerts a 
      marked proapoptotic effect, and (3) no evident relationships occur between the 
      effects of HBO on cell growth and ROS production.
FAU - Conconi, Maria Teresa
AU  - Conconi MT
AD  - Department of Pharmaceutical Sciences, University of Padua, Padua, Italy.
FAU - Baiguera, Silvia
AU  - Baiguera S
FAU - Guidolin, Diego
AU  - Guidolin D
FAU - Furlan, Claudio
AU  - Furlan C
FAU - Menti, Anna M
AU  - Menti AM
FAU - Vigolo, Simonetta
AU  - Vigolo S
FAU - Belloni, Anna S
AU  - Belloni AS
FAU - Parnigotto, Pier Paolo
AU  - Parnigotto PP
FAU - Nussdorfer, Gastone G
AU  - Nussdorfer GG
LA  - eng
PT  - Journal Article
PL  - England
TA  - J Investig Med
JT  - Journal of investigative medicine : the official publication of the American 
      Federation for Clinical Research
JID - 9501229
RN  - 0 (Reactive Oxygen Species)
RN  - S88TT14065 (Oxygen)
SB  - IM
MH  - 3T3 Cells
MH  - Animals
MH  - Apoptosis/*drug effects
MH  - Cell Division/drug effects
MH  - Clone Cells/drug effects
MH  - Fibroblasts/*drug effects/metabolism/pathology
MH  - *Hyperbaric Oxygenation
MH  - In Situ Nick-End Labeling
MH  - Mice
MH  - Oxygen/*toxicity
MH  - Reactive Oxygen Species/*metabolism
EDAT- 2003/08/22 05:00
MHDA- 2003/09/05 05:00
CRDT- 2003/08/22 05:00
PHST- 2003/08/22 05:00 [pubmed]
PHST- 2003/09/05 05:00 [medline]
PHST- 2003/08/22 05:00 [entrez]
AID - 10.1136/jim-51-04-24 [doi]
PST - ppublish
SO  - J Investig Med. 2003 Jul;51(4):227-32. doi: 10.1136/jim-51-04-24.