title: Improved neurological outcome by intramuscular injection of human amniotic fluid derived stem cells in a muscle denervation model.
abstract: OBJECTIVE
  The skeletal muscle develops various degrees of atrophy and metabolic dysfunction following nerve injury. Neurotrophic factors are essential for muscle regeneration. Human amniotic fluid derived stem cells (AFS) have the potential to secrete various neurotrophic factors necessary for nerve regeneration. In the present study, we assess the outcome of neurological function by intramuscular injection of AFS in a muscle denervation and nerve anastomosis model.
  
  
  METHODS
  Seventy two Sprague-Dawley rats weighing 200-250 gm were enrolled in this study. Muscle denervation model was conducted by transverse resection of a sciatic nerve with the proximal end sutured into the gluteal muscle. The nerve anastomosis model was performed by transverse resection of the sciatic nerve followed by four stitches reconnection. These animals were allocated to three groups: control, electrical muscle stimulation, and AFS groups.
  
  
  RESULTS
  NT-3 (Neurotrophin 3), BDNF (Brain derived neurotrophic factor), CNTF (Ciliary neurotrophic factor), and GDNF (Glia cell line derived neurotrophic factor) were highly expressed in AFS cells and supernatant of culture medium. Intra-muscular injection of AFS exerted significant expression of several neurotrophic factors over the distal end of nerve and denervated muscle. AFS caused high expression of Bcl-2 in denervated muscle with a reciprocal decrease of Bad and Bax. AFS preserved the muscle morphology with high expression of desmin and acetylcholine receptors. Up to two months, AFS produced significant improvement in electrophysiological study and neurological functions such as SFI (sciatic nerve function index) and Catwalk gait analysis. There was also significant preservation of the number of anterior horn cells and increased nerve myelination as well as muscle morphology.
  
  
  CONCLUSIONS
  Intramuscular injection of AFS can protect muscle apoptosis and likely does so through the secretion of various neurotrophic factors. This protection furthermore improves the nerve regeneration in a long term nerve anastomosis model.
authors.affiliation: Department of Medical Research, Taichung Veterans General Hospital, Taichung, Taiwan.
authors.forename: Chun-Jung
authors.initials: CJ
authors.lastname: Chen
authors.affiliation: Department of Medical Research, Taichung Veterans General Hospital, Taichung, Taiwan.
authors.forename: Fu-Chou
authors.initials: FC
authors.lastname: Cheng
authors.affiliation: Institute of Life Sciences, National Chung-Hsing University, Taichung, Taiwan.
authors.forename: Hong-Lin
authors.initials: HL
authors.lastname: Su
authors.affiliation: Institute of Biomedical Sciences, National Chung-Hsing University, Taichung, Taiwan.
authors.forename: Meei-Ling
authors.initials: ML
authors.lastname: Sheu
authors.affiliation: Institute of Biomedical Sciences, National Chung-Hsing University, Taichung, Taiwan.
authors.forename: Zong-Han
authors.initials: ZH
authors.lastname: Lu
authors.affiliation: Institute of Biomedical Sciences, National Chung-Hsing University, Taichung, Taiwan.
authors.forename: Chien-Yi
authors.initials: CY
authors.lastname: Chiang
authors.affiliation: Department of Neurosurgery, Chang Bing Show Chwan Memorial Hospital, Changhua, Taiwan.
authors.forename: Dar-Yu
authors.initials: DY
authors.lastname: Yang
authors.affiliation: Department of Neurosurgery, University of Virginia, Charlottesville, VA, United States of America.
authors.forename: Jason
authors.initials: J
authors.lastname: Sheehan
authors.affiliation: Department of Neurosurgery, Taichung Veterans General Hospital, Taichung, Taiwan; Faculty of Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan.
authors.forename: Hung-Chuan
authors.initials: HC
authors.lastname: Pan
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology*; D000714:Anastomosis, Surgical; D000818:Animals; D000870:Anterior Horn Cells / Q000502:physiology; D019208:Brain-Derived Neurotrophic Factor / Q000378:metabolism; D064987:Cell- and Tissue-Based Therapy / Q000379:methods; D020934:Ciliary Neurotrophic Factor / Q000378:metabolism; D003893:Desmin / Q000096:biosynthesis; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000378:metabolism; D006801:Humans; D007273:Injections, Intramuscular; D009121:Muscle Denervation; D018482:Muscle, Skeletal / Q000294:innervation; D009133:Muscular Atrophy / Q000628:therapy; D009414:Nerve Growth Factors / Q000378:metabolism*; D009416:Nerve Regeneration / Q000502:physiology*; D020933:Neurotrophin 3; D051381:Rats; D017207:Rats, Sprague-Dawley; D011950:Receptors, Cholinergic / Q000096:biosynthesis; D012584:Sciatic Nerve / Q000293:injuries / Q000378:metabolism; D020426:Sciatic Neuropathy / Q000503:physiopathology / Q000628:therapy*; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000378:metabolism; D014183:Transplantation, Heterologous; D051028:bcl-2-Associated X Protein / Q000378:metabolism; D051025:bcl-Associated Death Protein / Q000378:metabolism
chemical_list: C494821:Bad protein, rat; C494828:Bax protein, rat; D019208:Brain-Derived Neurotrophic Factor; D020934:Ciliary Neurotrophic Factor; D003893:Desmin; D051100:Glial Cell Line-Derived Neurotrophic Factor; C000595808:NTF3 protein, human; D009414:Nerve Growth Factors; D020933:Neurotrophin 3; D011950:Receptors, Cholinergic; D051028:bcl-2-Associated X Protein; D051025:bcl-Associated Death Protein; C579455:BDNF protein, human