PMID- 30092096
OWN - NLM
STAT- MEDLINE
DCOM- 20190131
LR  - 20190131
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 13
IP  - 8
DP  - 2018
TI  - High glucose-induced oxidative stress impairs proliferation and migration of 
      human gingival fibroblasts.
PG  - e0201855
LID - 10.1371/journal.pone.0201855 [doi]
LID - e0201855
AB  - Delayed gingival wound healing is widely observed in periodontal patients with 
      diabetes. However, the molecular mechanisms of the impaired function of gingival 
      fibroblasts in diabetes remain unclear. The purpose of this study was to 
      investigate changes in the properties of human gingival fibroblasts (HGFs) under 
      high-glucose conditions. Primary HGFs were isolated from healthy gingiva and 
      cultured with 5.5, 25, 50, and 75 mM glucose for 72 h. In vitro wound healing, 
      5-ethynyl-2'-deoxyuridine (EdU), and water-soluble tetrazolium salt (WST-8) 
      assays were performed to examine cell migration and proliferation. Lactase 
      dehydrogenase (LDH) levels were measured to determine cytotoxicity. The mRNA 
      expression levels of oxidative stress markers were quantified by real-time PCR. 
      Intracellular reactive oxygen species (ROS) were also measured in live cells. The 
      antioxidant N-acetyl-l-cysteine (NAC, 1 mM) was added to evaluate the involvement 
      of ROS in the glucose effect on HGFs. As a result, the in vitro wound healing 
      assay showed that high glucose levels significantly reduced fibroblast migration 
      and proliferation at 6, 12, 24, 36, and 48 h. The numbers of cells positive for 
      EdU staining were decreased, as was cell viability, at 50 and 75 mM glucose. A 
      significant increase in LDH was proportional to the glucose concentration. The 
      mRNA levels of heme oxygenase-1 and superoxide dismutase-1 and ROS levels were 
      significantly increased in HGFs after 72 h of exposure to 50 mM glucose 
      concentration. The addition of NAC diminished the inhibitory effect of high 
      glucose in the in vitro wound healing assay. The results of the present study 
      show that high glucose impairs the proliferation and migration of HGFs. 
      Fibroblast dysfunction may therefore be caused by high glucose-induced oxidative 
      stress and may explain the delayed gingival wound healing in diabetic patients.
FAU - Buranasin, Prima
AU  - Buranasin P
AD  - Department of Periodontology, Graduate School of Medical and Dental Sciences, 
      Tokyo Medical and Dental University (TMDU), Tokyo, Japan.
FAU - Mizutani, Koji
AU  - Mizutani K
AUID- ORCID: 0000-0002-1610-7430
AD  - Department of Periodontology, Graduate School of Medical and Dental Sciences, 
      Tokyo Medical and Dental University (TMDU), Tokyo, Japan.
FAU - Iwasaki, Kengo
AU  - Iwasaki K
AD  - Department of Nanomedicine, Graduate School of Medical and Dental Sciences, Tokyo 
      Medical and Dental University (TMDU), Tokyo, Japan.
FAU - Pawaputanon Na Mahasarakham, Chantida
AU  - Pawaputanon Na Mahasarakham C
AD  - Department of Restorative Dentistry, Faculty of Dentistry, Khon Kaen University, 
      Khon Kaen, Thailand.
FAU - Kido, Daisuke
AU  - Kido D
AD  - Department of Periodontology, Graduate School of Medical and Dental Sciences, 
      Tokyo Medical and Dental University (TMDU), Tokyo, Japan.
FAU - Takeda, Kohei
AU  - Takeda K
AD  - Department of Periodontology, Graduate School of Medical and Dental Sciences, 
      Tokyo Medical and Dental University (TMDU), Tokyo, Japan.
FAU - Izumi, Yuichi
AU  - Izumi Y
AD  - Department of Periodontology, Graduate School of Medical and Dental Sciences, 
      Tokyo Medical and Dental University (TMDU), Tokyo, Japan.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20180809
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Antioxidants)
RN  - 0 (RNA, Messenger)
RN  - 0 (Reactive Oxygen Species)
RN  - 0 (SOD1 protein, human)
RN  - EC 1.14.14.18 (HMOX1 protein, human)
RN  - EC 1.14.14.18 (Heme Oxygenase-1)
RN  - EC 1.15.1.1 (Superoxide Dismutase-1)
RN  - IY9XDZ35W2 (Glucose)
RN  - WYQ7N0BPYC (Acetylcysteine)
SB  - IM
MH  - Acetylcysteine/pharmacology
MH  - Adult
MH  - Aged
MH  - Antioxidants/pharmacology
MH  - Cell Movement/drug effects/*physiology
MH  - Cell Proliferation/drug effects/*physiology
MH  - Cell Survival/drug effects/physiology
MH  - Cells, Cultured
MH  - Female
MH  - Fibroblasts/drug effects/*metabolism/pathology
MH  - Gingiva/drug effects/injuries/*metabolism/pathology
MH  - Glucose/*adverse effects/metabolism
MH  - Heme Oxygenase-1/metabolism
MH  - Humans
MH  - Male
MH  - Middle Aged
MH  - Oxidative Stress/drug effects/*physiology
MH  - RNA, Messenger/metabolism
MH  - Reactive Oxygen Species/metabolism
MH  - Superoxide Dismutase-1/metabolism
MH  - Wound Healing/drug effects/physiology
PMC - PMC6084939
COIS- The authors have declared that no competing interests exist.
EDAT- 2018/08/10 06:00
MHDA- 2019/02/01 06:00
PMCR- 2018/08/09
CRDT- 2018/08/10 06:00
PHST- 2018/05/15 00:00 [received]
PHST- 2018/07/23 00:00 [accepted]
PHST- 2018/08/10 06:00 [entrez]
PHST- 2018/08/10 06:00 [pubmed]
PHST- 2019/02/01 06:00 [medline]
PHST- 2018/08/09 00:00 [pmc-release]
AID - PONE-D-18-14590 [pii]
AID - 10.1371/journal.pone.0201855 [doi]
PST - epublish
SO  - PLoS One. 2018 Aug 9;13(8):e0201855. doi: 10.1371/journal.pone.0201855. 
      eCollection 2018.