Extended magnetic resonance imaging studies on the effect of classically activated microglia transplantation on white matter regeneration following spinal cord focal injury in adult rats

Marcol,Wiesław; Ślusarczyk,Wojciech; Larysz‑Brysz,Magdalena; Łabuzek,Krzysztof; Kapustka,Bartosz; Staszkiewicz,Rafał; Rosicka,Paulina; Kalita,Katarzyna; Węglarz,Władysław; Lewin‑Kowalik,Joanna

doi:10.3892/etm.2017.5130

Extended magnetic resonance imaging studies on the effect of classically activated microglia transplantation on white matter regeneration following spinal cord focal injury in adult rats

Authors:
- Wiesław Marcol
- Wojciech Ślusarczyk
- Magdalena Larysz‑Brysz
- Krzysztof Łabuzek
- Bartosz Kapustka
- Rafał Staszkiewicz
- Paulina Rosicka
- Katarzyna Kalita
- Władysław Węglarz
- Joanna Lewin‑Kowalik
View Affiliations

Affiliations: Department of Physiology, School of Medicine in Katowice, Medical University of Silesia, 40‑752 Katowice, Poland, Department of Internal Medicine and Clinical Pharmacology, School of Medicine in Katowice, Medical University of Silesia, 40‑752 Katowice, Poland, Department of Magnetic Resonance Imaging, Institute of Nuclear Physics Polish Academy of Sciences, 31‑342 Kraków, Poland
Published online on: September 19, 2017 https://doi.org/10.3892/etm.2017.5130
Pages: 4869-4877
Copyright: © Marcol et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Spinal cord injuries are still a serious problem for regenerative medicine. Previous research has demonstrated that activated microglia accumulate in spinal lesions, influencing the injured tissues in various ways. Therefore, transplantation of activated microglia may have a beneficial role in the regeneration of the nervous system. The present study examined the influence of transplanted activated microglial cells in adult rats with injured spinal cords. Rats were randomly divided into an experimental (M) and control (C) group, and were subjected to non‑laminectomy focal injury of spinal cord white matter by means of a high‑pressured air stream. In group M, activated cultured microglial cells were injected twice into the site of injury. Functional outcome and morphological features of regeneration were analyzed during a 12‑week follow‑up. The lesions were characterized by means of magnetic resonance imaging (MRI). Neurons in the brain stem and motor cortex were labeled with FluoroGold (FG). A total of 12 weeks after surgery, spinal cords and brains were collected and subjected to histopathological and immunohistochemical examinations. Lesion sizes in the spinal cord were measured and the number of FG‑positive neurons was counted. Rats in group M demonstrated significant improvement of locomotor performance when compared with group C (P<0.05). MRI analysis demonstrated moderate improvement in water diffusion along the spinal cord in the group M following microglia treatment, as compared with group C. The water diffusion perpendicular to the spinal cord in group M was closer to the reference values for a healthy spinal cord than it was in group C. The sizes of lesions were also significantly smaller in group M than in the group C (P<0.05). The number of brain stem and motor cortex FG‑positive neurons in group M was significantly higher than in group C. The present study demonstrated that delivery of activated microglia directly into the injured spinal cord gives some positive effects for the regeneration of the white matter.

View Figures

View References

1	Schwartz M, Cohen I, Lazarov-Spiegler O, Moalem G and Yoles E: The remedy may lie in ourselves: Prospects for immune cell therapy in central nervous system protection and repair. J Mol Med (Berl). 77:713–717. 1999. View Article : Google Scholar : PubMed/NCBI
2	Schwartz M: Macrophages and microglia in central nervous system injury: Are they helpful or harmful? J Cereb Blood Flow Metab. 23:385–394. 2003. View Article : Google Scholar : PubMed/NCBI
3	Moalem G, Leibowitz-Amit R, Yoles E, Mor F, Cohen IR and Schwartz M: Autoimmune T cells protect neurons from secondary degeneration after central nervous system axotomy. Nat Med. 5:49–55. 1999. View Article : Google Scholar : PubMed/NCBI
4	Loane DJ and Byrnes KR: Role of microglia in neurotrauma. Neurotherapeutics. 7:366–377. 2010. View Article : Google Scholar : PubMed/NCBI
5	Del Río Hortega P: La neuroglía normal. Conceptos de neurogliona y angiogliona. Arch Histol Norm Patol. 1:5–71. 1942.
6	Walker PA, Shah SK, Jimenez F, Aroom KR, Harting MT and Cox CS Jr: Bone marrow-derived stromal cell therapy for traumatic brain injury is neuroprotective via stimulation of non-neurologic organ systems. Surgery. 152:790–793. 2012. View Article : Google Scholar : PubMed/NCBI
7	Miyake T, Tsuchihashi Y, Kitamura and Fujita S: Immunohistochemical studies of blood monocytes infiltrating into the neonatal rat brain. Acta Neuropathol. 62:291–297. 1984. View Article : Google Scholar : PubMed/NCBI
8	Perry VH, Hume DA and Gordon S: Immunohistochemical localization of macrophages and microglia in the adult and developing mouse brain. Neuroscience. 15:313–326. 1985. View Article : Google Scholar : PubMed/NCBI
9	David S and Kroner A: Repertoire of microglial and macrophage responses after spinal cord injury. Nat Rev Neurosci. 12:388–399. 2011. View Article : Google Scholar : PubMed/NCBI
10	Fujita S and Kitamura T: Origin of brain macrophages and the nature of the so-called microglia. Acta Neuropathol Suppl. Suppl 6:S291–S296. 1975.
11	Li Y, Li D and Raisman G: Transplanted schwann cells, not olfactory ensheathing cells, myelinate optic nerve fibres. Glia. 55:312–316. 2007. View Article : Google Scholar : PubMed/NCBI
12	Aloisi F: Immune function of microglia. Glia. 36:165–179. 2001. View Article : Google Scholar : PubMed/NCBI
13	Yaguchi M, Ohta S, Toyama Y, Kawakami Y and Toda M: Functional recovery after spinal cord injury in mice through activation of microglia and dendritic cells after IL-12 administration. J Neurosci Res. 86:1972–1980. 2008. View Article : Google Scholar : PubMed/NCBI
14	Marcol W, Slusarczyk W, Gzik M, Larysz-Brysz M, Bobrowski M, Grynkiewicz-Bylina B, Rosicka P, Kalita K, Węglarz W, Barski JJ, et al: Air gun impactor-a novel model of graded white matter spinal cord injury in rodents. J Reconstr Microsurg. 28:561–568. 2012. View Article : Google Scholar : PubMed/NCBI
15	Labuzek K, Kowalski J, Gabryel B and Herman ZS: Chlorpromazine and loxapine reduce interleukin-1beta and interleukin-2 release by rat mixed glial and microglial cell cultures. Eur Neuropsychopharmacol. 15:23–30. 2005. View Article : Google Scholar : PubMed/NCBI
16	Mosmann T: Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Methods. 65:55–63. 1983. View Article : Google Scholar : PubMed/NCBI
17	Satoh J, Lee YB and Kim SU: T-cell costimulatory molecules B7-1 (CD80) and B7-2 (CD86) are expressed in human microglia but not in astrocytes in culture. Brain Res. 704:92–96. 1995. View Article : Google Scholar : PubMed/NCBI
18	Giri S, Nath N, Smith B, Viollet B, Singh AK and Singh I: 5-aminoimidazole-4-carboxamide-1-beta-4-ribofuranoside inhibits proinflammatory response in glial cells: A possible role of AMP-activated protein kinase. J Neurosci. 24:479–487. 2004. View Article : Google Scholar : PubMed/NCBI
19	Basso DM, Beattie MS and Bresnahan JC: A sensitive and reliable locomotor rating scale for open field testing in rats. J Neurotrauma. 12:1–21. 1995. View Article : Google Scholar : PubMed/NCBI
20	Karimi-Abdolrezaee S, Eftekharpour E, Wang J, Morshead CM and Fehlings MG: Delayed transplantation of adult neural precursor cells promotes remyelination and functional neurological recovery after spinal cord injury. J Neurosci. 26:3377–3389. 2006. View Article : Google Scholar : PubMed/NCBI
21	Węglarz WP, Jasiński A, Krzyżak AT, Kozłowski P, Adamek D, Sagnowski P and Pindel J: MR microscopy of water diffusion tensor in biological systems. Appl Magn Reson. 15:333–341. 1998. View Article : Google Scholar
22	Weglarz WP, Adamek D, Markiewicz J, Skórka T, Kulinowski P and Jasiński A: Analysis of the diffusion weighted MR microscopy data of excised spinal cord of a rat on the basis of model of restricted diffusion. Solid State Nucl Magn Reson. 25:88–93. 2004. View Article : Google Scholar : PubMed/NCBI
23	Minati L and Węglarz WP: Physical, foundations, models, and methods of diffusion magnetic resonance imaging of the brain: A review. Concepts Magnetic Resonance. 30A:278–307. 2007. View Article : Google Scholar
24	Knoller N, Auerbach G, Fulga V, Zelig G, Attias J, Bakimer R, Marder JB, Yoles E, Belkin M, Schwartz M and Hadani M: Clinical experience using incubated autologous macrophages as a treatment for complete spinal cord injury: Phase I study results. J Neurosurg Spine. 3:173–181. 2005. View Article : Google Scholar : PubMed/NCBI
25	Fujimoto Y, Yamasaki T, Tanaka N, Mochizuki Y, Kajihara H, Ikuta Y and Ochi M: Differential activation of astrocytes and microglia after spinal cord injury in the fetal rat. Eur Spine J. 15:223–233. 2006. View Article : Google Scholar : PubMed/NCBI
26	Prewitt CM, Niesman IR, Kane CJ and Houlé JD: Activated macrophage/microglial cells can promote the regeneration of sensory axons into the injured spinal cord. Exp Neurol. 148:433–443. 1997. View Article : Google Scholar : PubMed/NCBI
27	Hynds DL, Rangappa N, Ter Beest J, Snow DM and Rabchevsky AG: Microglia enhance dorsal root ganglion outgrowth in Schwann cell cultures. Glia. 46:218–223. 2004. View Article : Google Scholar : PubMed/NCBI
28	Yu TB, Cheng YS, Zhao P, Kou DW, Sun K, Chen BH and Wang AM: Immune therapy with cultured microglia grafting into the injured spinal cord promoting the recovery of rat's hind limb motor function. Chin J Traumatol. 12:291–295. 2009.PubMed/NCBI
29	Mukaino M, Nakamura M, Yamada O, Okada S, Morikawa S, Renault-Mihara F, Iwanami A, Ikegami T, Ohsugi Y, Tsuji O, et al: Anti-IL-6-receptor antibody promotes repair of spinal cord injury by inducing microglia-dominant inflammation. Exp Neurol. 224:403–414. 2010. View Article : Google Scholar : PubMed/NCBI
30	Morino T, Ogata T, Takeba J and Yamamoto H: Microglia inhibition is a target of mild hypothermic treatment after the spinal cord injury. Spinal Cord. 46:425–431. 2008. View Article : Google Scholar : PubMed/NCBI
31	Wu B, Matic D, Djogo N, Szpotowicz E, Schachner M and Jakovcevski I: Improved regeneration after spinal cord injury in mice lacking functional T- and B-lymphocytes. Exp Neurol. 237:274–285. 2012. View Article : Google Scholar : PubMed/NCBI
32	Emmetsberger J and Tsirka SE: Microglial inhibitory factor (MIF/TKP) mitigates secondary damage following spinal cord injury. Neurobiol Dis. 47:295–309. 2012. View Article : Google Scholar : PubMed/NCBI
33	Klusman I and Schwab ME: Effects of pro-inflammatory cytokines in experimental spinal cord injury. Brain Res. 762:173–184. 1997. View Article : Google Scholar : PubMed/NCBI