PMID- 10779676
OWN - NLM
STAT- MEDLINE
DCOM- 20000630
LR  - 20190818
IS  - 0304-3959 (Print)
IS  - 0304-3959 (Linking)
VI  - 86
IP  - 1-2
DP  - 2000 May
TI  - Lumbar transplant of neurons genetically modified to secrete brain-derived 
      neurotrophic factor attenuates allodynia and hyperalgesia after sciatic nerve 
      constriction.
PG  - 195-210
AB  - Chronic delivery of anti-nociceptive molecules by means of cell grafts near the 
      pain processing centers of the spinal cord is a newly developing technique for 
      the treatment of neuropathic pain. The rat neuronal cell line, RN33B, derived 
      from E13 rat brainstem raphe and immortalized with the SV40 temperature-sensitive 
      allele of large T antigen (tsTag), was transfected with rat brain-derived 
      neurotrophic factor cDNA (BDNF), and the BDNF-synthesizing cell line, 33BDNF.4, 
      was isolated. The 33BDNF.4 cells synthesized mature BDNF protein at permissive 
      temperature (33 degrees C), when the cells were proliferating, and during 
      differentiation at non-permissive temperature (39 degrees C) in vitro. The 
      bio-active BDNF protein was also secreted by the cells during both growth 
      conditions, as measured by ELISA analysis of BDNF content and secretion. The 
      bio-activity of the BDNF in 33BDNF.4 cell conditioned media was assessed by 
      neurite outgrowth from E15 dorsal root ganglion (DRG) cultures. A control cell 
      line, 33V1, transfected with the vector alone, did not synthesize or secrete any 
      significant BDNF at either growth condition. Both cell lines were used as grafts 
      in a model of chronic neuropathic pain induced by unilateral chronic constriction 
      injury (CCI) of the sciatic nerve. Pain-related behaviors, including cold and 
      tactile allodynia and thermal and tactile hyperalgesia, were evaluated after CCI 
      in the affected hindpaw. When 33BDNF.4 and 33V1 cells were transplanted in the 
      lumbar subarachnoid space of the spinal cord 1 week after CCI, they survived 
      greater than 7 weeks on the pia mater around the spinal cord and the 33BDNF.4 
      cells continued to synthesize BDNF in vivo. Furthermore, the tactile and cold 
      allodynia and tactile and thermal hyperalgesia induced by CCI was significantly 
      reduced during the 2-7 week period after grafts of 33BDNF.4 cells. The maximal 
      effect on chronic pain behaviors with the BDNF grafts occurred 2-3 weeks after 
      transplant and the anti-nociceptive effects of the BDNF cell grafts was 
      permanent. Transplants of the control 33V1 cells had no effect on the allodynia 
      and hyperalgesia induced by CCI and these cells did not synthesize BDNF in vivo. 
      These data suggest that a chronically applied, low local dose of BDNF supplied by 
      transplanted cells near the spinal dorsal horn was able to reverse the 
      development of chronic neuropathic pain following CCI. The use of neural cell 
      lines that are able to deliver anti-nociceptive molecules, such as BDNF, in a 
      model of chronic pain offers a novel approach to pain management and such 
      'biologic minipumps' can be developed for safe use in humans.
FAU - Cejas, P J
AU  - Cejas PJ
AD  - The Miami Project to Cure Paralysis, University of Miami School of Medicine, FL 
      33136, USA.
FAU - Martinez, M
AU  - Martinez M
FAU - Karmally, S
AU  - Karmally S
FAU - McKillop, M
AU  - McKillop M
FAU - McKillop, J
AU  - McKillop J
FAU - Plunkett, J A
AU  - Plunkett JA
FAU - Oudega, M
AU  - Oudega M
FAU - Eaton, M J
AU  - Eaton MJ
LA  - eng
GR  - 36438-03/PHS HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Pain
JT  - Pain
JID - 7508686
RN  - 0 (Brain-Derived Neurotrophic Factor)
SB  - IM
MH  - Animals
MH  - Behavior, Animal/physiology
MH  - Brain-Derived Neurotrophic Factor/*genetics/*metabolism
MH  - Cell Line
MH  - Cell Transplantation/*physiology
MH  - Clone Cells
MH  - Cold Temperature
MH  - Enzyme-Linked Immunosorbent Assay
MH  - Ganglia, Spinal/cytology/metabolism
MH  - Graft Survival
MH  - Hot Temperature
MH  - Hyperalgesia/*therapy
MH  - Immunohistochemistry
MH  - Ligation
MH  - Mice
MH  - Neurons/*metabolism/*transplantation
MH  - Pain/psychology
MH  - *Pain Management
MH  - Pain Measurement
MH  - Physical Stimulation
MH  - Rats
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Sciatic Neuropathy/*therapy
MH  - Transfection/genetics
EDAT- 2000/04/26 09:00
MHDA- 2000/07/08 11:00
CRDT- 2000/04/26 09:00
PHST- 2000/04/26 09:00 [pubmed]
PHST- 2000/07/08 11:00 [medline]
PHST- 2000/04/26 09:00 [entrez]
AID - S0304-3959(00)00245-1 [pii]
AID - 10.1016/s0304-3959(00)00245-1 [doi]
PST - ppublish
SO  - Pain. 2000 May;86(1-2):195-210. doi: 10.1016/s0304-3959(00)00245-1.