PMID- 22410147
OWN - NLM
STAT- MEDLINE
DCOM- 20121130
LR  - 20151119
IS  - 1879-1506 (Electronic)
IS  - 0003-9969 (Linking)
VI  - 57
IP  - 9
DP  - 2012 Sep
TI  - Application of eGFP to label human periodontal ligament stem cells in periodontal 
      tissue engineering.
PG  - 1241-50
LID - 10.1016/j.archoralbio.2012.02.017 [doi]
AB  - OBJECTIVES: To establish human periodontal ligament stem cells (hPDLSC) with high 
      and stable expression of enhanced green fluorescent protein (eGFP) and to obtain 
      an ideal vector expression system that suitable for gene therapy in periodontal 
      tissue engineering. METHODS: hPDLSCs were transfected with eGFP for 48h via 
      different MOI (25, 50, 100, 200 and 400) by lentiviral vector, the transfection 
      efficiency was evaluated by fluorescent microscopy and flow cytometry, and 
      transfected hPDLSCs proliferation was evaluated by MTT. Pluripotent, 
      differentiation capacity and ALP expression status were determined further. 
      Osteoblast-associated genes expressions for osteogenesis were evaluated by 
      quantitative-PCR. In addition, rat molar periodontal fenestration defect model 
      was used for evaluating periodontal tissue engineering. RESULTS: The transfection 
      efficiency after 48h were 44.7%, 60.9%, 71.7%, 85.8%, and 86.9% respectively. 
      There was no significant effect of transfection (at different MOI levels of 25, 
      50, 100, and 200) on the proliferation of hPDLSCs (designated as eGFP-hPDLSCs) 
      compared with hPDLSCs (P>0.05). However, proliferation of eGFP hPDLSCs at MOI 400 
      became slower (P<0.05). Both eGFP hPDLSCs and hPDLSCs were able to differentiate 
      into osteocytes and adipocytes under certain conditioned media. At 7 days, 
      expression levels of COL-1, RUNX2 in hPDLSCS were higher than those in eGFP 
      hPDLSCs (P<0.05); expression levels of ALP and OPN in eGFP hPDLSCs were similar 
      to those in hPDLSCs (P>0.05). Newly regenerated bone formation was observed in 
      the defect model used. CONCLUSIONS: Among the transfection conditions, 48h 
      transfection at MOI 200 is optimal for labelling hPDLSCs with eGFP in a 
      lentiviral vector. There is no change in capability of the eGFP hPDLSCs 
      osteogenesis. The lentiviral vector with eGFP is an appropriate expression vector 
      system and hPDLSCs are ideal seeding cells for gene therapy in periodontal tissue 
      engineering.
CI  - Copyright (c) 2012 Elsevier Ltd. All rights reserved.
FAU - Wen, Yong
AU  - Wen Y
AD  - School of Stomatology, Shandong University, No. 44-1 Wenhua Xi Road, Jinan, 
      Shandong, PR China. wenyongsdu@yahoo.com.cn
FAU - Lan, Jing
AU  - Lan J
FAU - Huang, Haiyun
AU  - Huang H
FAU - Yu, Meijiao
AU  - Yu M
FAU - Cui, Jun
AU  - Cui J
FAU - Liang, Jin
AU  - Liang J
FAU - Jiang, Baoqi
AU  - Jiang B
FAU - Xu, Xin
AU  - Xu X
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20120310
PL  - England
TA  - Arch Oral Biol
JT  - Archives of oral biology
JID - 0116711
RN  - 0 (Collagen Type I)
RN  - 0 (Core Binding Factor Alpha 1 Subunit)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Osteoprotegerin)
RN  - 0 (RUNX2 protein, human)
RN  - 0 (TNFRSF11B protein, human)
RN  - 0 (enhanced green fluorescent protein)
RN  - 147336-22-9 (Green Fluorescent Proteins)
RN  - EC 3.1.3.1 (Alkaline Phosphatase)
SB  - IM
MH  - Adipocytes/physiology
MH  - Adipogenesis/physiology
MH  - Alkaline Phosphatase/analysis
MH  - Alveolar Bone Loss/surgery
MH  - Animals
MH  - Bone Regeneration/physiology
MH  - Cell Culture Techniques
MH  - Cell Differentiation/physiology
MH  - Cell Proliferation
MH  - Collagen Type I/analysis
MH  - Core Binding Factor Alpha 1 Subunit/analysis
MH  - Disease Models, Animal
MH  - *Fluorescent Dyes
MH  - *Genetic Therapy
MH  - Genetic Vectors/genetics
MH  - *Green Fluorescent Proteins
MH  - Humans
MH  - Lentivirus/genetics
MH  - Osteoblasts/cytology
MH  - Osteocytes/physiology
MH  - Osteogenesis/physiology
MH  - Osteoprotegerin
MH  - Periodontal Ligament/*cytology
MH  - Pluripotent Stem Cells/cytology
MH  - Rats
MH  - Rats, Sprague-Dawley
MH  - Stem Cells/*cytology
MH  - *Tissue Engineering
MH  - Transfection
EDAT- 2012/03/14 06:00
MHDA- 2012/12/10 06:00
CRDT- 2012/03/14 06:00
PHST- 2011/11/04 00:00 [received]
PHST- 2012/02/07 00:00 [revised]
PHST- 2012/02/13 00:00 [accepted]
PHST- 2012/03/14 06:00 [entrez]
PHST- 2012/03/14 06:00 [pubmed]
PHST- 2012/12/10 06:00 [medline]
AID - S0003-9969(12)00065-9 [pii]
AID - 10.1016/j.archoralbio.2012.02.017 [doi]
PST - ppublish
SO  - Arch Oral Biol. 2012 Sep;57(9):1241-50. doi: 10.1016/j.archoralbio.2012.02.017. 
      Epub 2012 Mar 10.