PMID- 20210582
OWN - NLM
STAT- MEDLINE
DCOM- 20100602
LR  - 20100309
IS  - 1746-076X (Electronic)
IS  - 1746-0751 (Linking)
VI  - 5
IP  - 2
DP  - 2010 Mar
TI  - In vitro study of stem cell communication via gap junctions for fibrocartilage 
      regeneration at entheses.
PG  - 221-9
LID - 10.2217/rme.09.86 [doi]
AB  - BACKGROUND: Entheses are fibrocartilaginous organs that bridge ligament with bone 
      at their interface and add significant insertional strength. To replace a 
      severely damaged ligament, a tissue-engineered graft preinstalled with 
      interfacial fibrocartilage, which is being regenerated from stem cells, appears 
      to be more promising than ligament-alone graft. Such a concept can be realized by 
      a biomimetic approach of establishing a dynamic communication of stem cells with 
      bone cells and/or ligament fibroblasts in vitro. AIM: The current study has two 
      objectives. The first objective is to demonstrate functional coculture of bone 
      marrow-derived stem cells (BMSCs) with mature bone cells/ligament fibroblasts as 
      evidenced by gap-junctional communication in vitro. The second objective is to 
      investigate the role of BMSCs in the regeneration of fibrocartilage within the 
      coculture. MATERIALS & METHODS: Rabbit bone/ligament fibroblasts were 
      dual-stained with DiI-Red and calcein (gap-junction permeable dye), and 
      cocultured with unlabeled BMSCs at fixed ratio (1:10). The functional gap 
      junction was demonstrated by the transfer of calcein from donor to recipient 
      cells that was confirmed and quantified by flow cytometry. Type 2 collagen 
      (cartilage extracellular matrix-specific protein) expressed by the mixed cell 
      lines in the cocultures were estimated by real-time reverse transcription PCR and 
      compared with that of the ligament-bone coculture (control). RESULTS: Significant 
      transfer of calcein into BMSCs was observed and flow cytometry analyses showed a 
      gradual increase in the percentage of BMSCs acquiring calcein with time. 
      Cocultures that included BMSCs expressed significantly more type 2 collagen 
      compared with the control. CONCLUSION: The current study, for the first time, 
      reported the expression of gap-junctional communication of BMSCs with two 
      adherent cell lines of musculoskeletal system in vitro and also confirmed that 
      incorporation of stem cells augments fibrocartilage regeneration. The results 
      open up a path to envisage a composite graft preinstalled with enthesial 
      fibrocartilage using a stem cell-based coculture system.
FAU - Nayak, Bibhukalyan Prasad
AU  - Nayak BP
AD  - Department of Biotechnology & Medical Engineering, National Institute of 
      Technology, Rourkela, India. crl.bpnayak@gmail.com
FAU - Goh, James Cho Hong
AU  - Goh JC
FAU - Toh, Siew Lok
AU  - Toh SL
FAU - Satpathy, Gyan Ranjan
AU  - Satpathy GR
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Regen Med
JT  - Regenerative medicine
JID - 101278116
RN  - 0 (Collagen Type II)
RN  - 0 (Coloring Agents)
SB  - IM
MH  - Animals
MH  - Bone Marrow Cells/cytology/metabolism
MH  - *Cell Communication
MH  - Cell Lineage
MH  - Coculture Techniques
MH  - Collagen Type II/genetics/metabolism
MH  - Coloring Agents/metabolism
MH  - Fibrocartilage/cytology/*physiology
MH  - Flow Cytometry
MH  - Gap Junctions/*metabolism
MH  - Gene Expression Regulation
MH  - Ligaments/cytology/*physiology
MH  - Male
MH  - Microscopy, Fluorescence
MH  - Rabbits
MH  - Regeneration/*physiology
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Stem Cells/*cytology/metabolism
EDAT- 2010/03/10 06:00
MHDA- 2010/06/03 06:00
CRDT- 2010/03/10 06:00
PHST- 2010/03/10 06:00 [entrez]
PHST- 2010/03/10 06:00 [pubmed]
PHST- 2010/06/03 06:00 [medline]
AID - 10.2217/rme.09.86 [doi]
PST - ppublish
SO  - Regen Med. 2010 Mar;5(2):221-9. doi: 10.2217/rme.09.86.