PMID- 25945496 OWN - NLM STAT- MEDLINE DCOM- 20160425 LR - 20201209 IS - 1932-6203 (Electronic) IS - 1932-6203 (Linking) VI - 10 IP - 5 DP - 2015 TI - Improved neurological outcome by intramuscular injection of human amniotic fluid derived stem cells in a muscle denervation model. PG - e0124624 LID - 10.1371/journal.pone.0124624 [doi] LID - e0124624 AB - PURPOSE: 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. MATERIALS AND 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. CONCLUSION: 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. FAU - Chen, Chun-Jung AU - Chen CJ AD - Department of Medical Research, Taichung Veterans General Hospital, Taichung, Taiwan. FAU - Cheng, Fu-Chou AU - Cheng FC AD - Department of Medical Research, Taichung Veterans General Hospital, Taichung, Taiwan. FAU - Su, Hong-Lin AU - Su HL AD - Institute of Life Sciences, National Chung-Hsing University, Taichung, Taiwan. FAU - Sheu, Meei-Ling AU - Sheu ML AD - Institute of Biomedical Sciences, National Chung-Hsing University, Taichung, Taiwan. FAU - Lu, Zong-Han AU - Lu ZH AD - Institute of Biomedical Sciences, National Chung-Hsing University, Taichung, Taiwan. FAU - Chiang, Chien-Yi AU - Chiang CY AD - Institute of Biomedical Sciences, National Chung-Hsing University, Taichung, Taiwan. FAU - Yang, Dar-Yu AU - Yang DY AD - Department of Neurosurgery, Chang Bing Show Chwan Memorial Hospital, Changhua, Taiwan. FAU - Sheehan, Jason AU - Sheehan J AD - Department of Neurosurgery, University of Virginia, Charlottesville, VA, United States of America. FAU - Pan, Hung-Chuan AU - Pan HC AD - Department of Neurosurgery, Taichung Veterans General Hospital, Taichung, Taiwan; Faculty of Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20150506 PL - United States TA - PLoS One JT - PloS one JID - 101285081 RN - 0 (Bad protein, rat) RN - 0 (Bax protein, rat) RN - 0 (Brain-Derived Neurotrophic Factor) RN - 0 (Ciliary Neurotrophic Factor) RN - 0 (Desmin) RN - 0 (Glial Cell Line-Derived Neurotrophic Factor) RN - 0 (NTF3 protein, human) RN - 0 (Nerve Growth Factors) RN - 0 (Neurotrophin 3) RN - 0 (Receptors, Cholinergic) RN - 0 (bcl-2-Associated X Protein) RN - 0 (bcl-Associated Death Protein) RN - 7171WSG8A2 (BDNF protein, human) SB - IM MH - Amniotic Fluid/*cytology MH - Anastomosis, Surgical MH - Animals MH - Anterior Horn Cells/physiology MH - Brain-Derived Neurotrophic Factor/metabolism MH - Cell- and Tissue-Based Therapy/methods MH - Ciliary Neurotrophic Factor/metabolism MH - Desmin/biosynthesis MH - Glial Cell Line-Derived Neurotrophic Factor/metabolism MH - Humans MH - Injections, Intramuscular MH - Muscle Denervation MH - Muscle, Skeletal/innervation MH - Muscular Atrophy/therapy MH - Nerve Growth Factors/*metabolism MH - Nerve Regeneration/*physiology MH - Neurotrophin 3 MH - Rats MH - Rats, Sprague-Dawley MH - Receptors, Cholinergic/biosynthesis MH - Sciatic Nerve/injuries/metabolism MH - Sciatic Neuropathy/physiopathology/*therapy MH - *Stem Cell Transplantation MH - Stem Cells/metabolism MH - Transplantation, Heterologous MH - bcl-2-Associated X Protein/metabolism MH - bcl-Associated Death Protein/metabolism PMC - PMC4422615 COIS- Competing Interests: The authors have declared that no competing interests exist. EDAT- 2015/05/07 06:00 MHDA- 2016/04/26 06:00 PMCR- 2015/05/06 CRDT- 2015/05/07 06:00 PHST- 2013/11/21 00:00 [received] PHST- 2015/03/17 00:00 [accepted] PHST- 2015/05/07 06:00 [entrez] PHST- 2015/05/07 06:00 [pubmed] PHST- 2016/04/26 06:00 [medline] PHST- 2015/05/06 00:00 [pmc-release] AID - PONE-D-13-48494 [pii] AID - 10.1371/journal.pone.0124624 [doi] PST - epublish SO - PLoS One. 2015 May 6;10(5):e0124624. doi: 10.1371/journal.pone.0124624. eCollection 2015.