In vitro model of the glial scar

² State Key Laboratory for Manufacturing System Engineering, School of Mechanical Engineering, Xi’an Jiaotong University, Xi’an, ShaanXi 710054, China

³ Department of Neurosurgery, Xijing Hospital, Fourth Military Medical University, Xi’an, Shaanxi Province 710032, China

⁴ Departamento de Ingeniería Mecánica, Universidad de Las Palmas de Gran Canaria, 35017 Las Palmas de Gran Canaria, Spain

IJB 2019, 5(2), 235 https://doi.org/10.18063/ijb.v5i2.235

Published: 30 July 2019

© 2019 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )

Download PDF

Cite

XML

Abstract

The trauma of central nervous system (CNS) can lead to glial scar, and it can limit the regeneration of neurons at the injured area, which is considered to be a major factor affecting the functional recovery of patients with CNS injury. At present, the study of the glial scar model in vitro is still limited to two-dimensional culture, and the state of the scar in vivo cannot be well mimicked. Therefore, we use a collagen gel and astrocytes to construct a three-dimensional (3D) model in vitro to mimic natural glial scar tissue. The effects of concentration changes of astrocytes on cell morphology, proliferation, and tissue performance were investigated. After 8 days of culture in vitro, the results showed that the tissue model contracted, with a measured shrinkage rate of 4.5%, and the compressive elastic modulus increased to nearly 4 times. Moreover, the astrocytes of the 3D tissue model have the ability of proliferation, hyperplasia, and formation of scar clusters. It indicates that the model we constructed has the characteristics of glial scar tissue to some extent and can provide an in vitro model for the research of glial scar and brain diseases.

Keywords

Glial scar

In vitro

Three-dimensional

References

1. Kawano H, Kimura-Kuroda J, Komuta Y, et al., 2012, Role of the Lesion Scar in the Response to Damage and Repair of the Central Nervous System. Cell Tissue Res, 349:169-80. DOI 10.1007/s00441-012-1336-5.

2. Aand JM, Berry M, 1985, Observation on Astrocyte Response to a Cerebral Stab Wound in Adult Rats. Brain Res, 327:61-9.

3. Maxwell W, Follows R, Ashhurst D, et al., 1990, The Response of the Cerebral Hemisphere of the Rat to Injury. I. The Mature Rat. Philos Trans R Soc Lond B Biol Sci, 328:479- 500. DOI 10.1098/rstb.1990.0121.

4. Mand CS, Fawcett J, 2001, The Astrocyte/Meningeal Cell Interface a Barrier to Successful Nerve Regeneration? Cell Tissue Res, 305:267-73. DOI 10.1007/s004410100384.

5. Yoshioka N, Kimura-Kuroda J, Saito T, et al., 2011, Small Molecule Inhibitor of Type I Transforming Growth Factorbeta Receptor Kinase Ameliorates the Inhibitory Milieu in Injured Brain and Promotes Regeneration of Nigrostriatal Dopaminergic Axons. J Neurosci Res, 89(3):381-93. DOI 10.1002/jnr.22552.

6. Yoshioka N, Hisanaga S, Kawano H, 2010, Suppression of Fibrotic Scar Formation Promotes Axonal Regeneration Without Disturbing Blood Brain Barrier Repair and Withdrawal of Leukocytes After Traumatic Brain Injury. J Comp Neurol, 518:3867-81. DOI 10.1002/cne.22431.

7. Mand P, Nilsson M, 2005, Astrocyte Activation and Reactive Gliosis. Glia, 50:427-34.

8. Anderson MF, Blomstrand F, Blomstrand C, et al., 2003, Astrocytes and Stroke: Networking for Survival? Neurochem Res, 28(2):293-305. DOI 10.1023/a:1022385402197.

9. Tatsumi K, Haga S, Matsuyoshi H, et al., 2005, Characterization of Cells with Proliferative Activity after a Brain Injury. Neurochem Int, 46(5):381-9. DOI 10.1016/j.neuint.2004.12.007.

10. Norenberg MD, 1994, Astrocyte Responses to CNS Injury. J Neuropathol Exp Neurol, 53(3):213-20.

11. Gao K, Wang CR, Jiang F, et al., 2013, Traumatic Scratch Injury in Astrocytes Triggers Calcium Influx to Activate the JNK/c-Jun/AP-1 Pathway and Switch on GFAP Expression. Glia, 61(12):2063-77. DOI 10.1002/glia.22577.

12. Sofroniew MV, 2015, Astrogliosis. Cold Spring Harb Perspect Biol, 7(2):a20420.

13. Kozai TDY, Jaquins-Gerstl AS, Vazquez AL, et al., 2015, Brain Tissue Responses to Neural Implants Impact Signal Sensitivity and Intervention Strategies. ACS Chem Neurosci, 6(1):48-67. DOI 10.1021/cn500256e.

14. Woeppel K, Qand Y, Cui XT, 2017, Recent Advances in Neural Electrode-Tissue Interfaces. Curr Opin Biomed Eng, 4:21-31.

15. Chen N, Luo B, Yang IH, et al., 2018, Biofunctionalized Platforms Towards Long-term Neural Interface. Curr Opin Biomed Eng, 6:81-91.

16. Jand S, Miller JH, 2014, Regeneration Beyond the Glial Scar. Exp Neurol, 253(1):197-207.

17. Rand FA, Silver J, 2016, Targeting Astrocytes in CNS Injury and Disease: A Translational Research Approach. Prog Neurobiol, 144:173-87.

18. Zhan JS, Gao K, Chai RC, et al., 2017, Astrocytes in Migration. Neurochem Res, 42(1):272-82.

19. Yu AC, Lee YL, Eng LF, 1993, Astrogliosis in Culture: I. The Model and the Effect of Antisense Oligonucleotides on Glial Fibrillary Acidic Protein Synthesis. J Neurosci Res, 34(3):295-303. DOI 10.1002/jnr.490340306.

20. Kimura-Kuroda J, Teng X, Komuta Y, et al., 2010, An in vitro Model of the Inhibition of Axon Growth in the Lesion Scar Formed after Central Nervous System Injury. Mol Cell Neurosci, 43(2):177-87. DOI 10.1016/j.mcn.2009.10.008.

21. Spencer KC, Sy JC, Falcón-Banchs R, et al., 2017, A Three Dimensional in vitro Glial Scar Model to Investigate the Local Strain Effects from Micromotion Around Neural Implants. Lab A Chip, 17(5):795-804. DOI 10.1039/c6lc01411a.

22. Rocha DN, Ferraz-Nogueira JP, Barrias CC, et al., 2015, Extracellular Environment Contribution to Astrogliosis Lessons Learned from a Tissue Engineered 3D Model of the Glial Scar. Front Cell Neurosci, 9(1):377. DOI 10.3389/fncel.2015.00377.

23. García-Fernández L, Halstenberg S, Unger RE, et al., 2010, Anti-Angiogenic Activity of Heparin-like Polysulfonated Polymeric Drugs in 3D Human Cell Culture. Biomaterials, 31(31):7863-72. DOI 10.1016/j.biomaterials.2010.07.022.

24. Jand S, Pompe T, 2018, Biomimetic Tumor Microenvironments Based on Collagen Matrices. Biomater Sci, 6:2009-24. DOI 10.1039/c8bm00303c.

25. Bell E, Ivarsson B, Merrill C, 1979, Production of a Tissue-like Structure by Contraction of Collagen Lattices by Human Fibroblasts of Different Proliferative Potential in vitro. Proc Natl Acad Sci U S A, 76(3):1274-8. DOI 10.1073/pnas.76.3.1274.

26. Lázaro J, Dechmann DKN, Lapoint S, et al., 2017, Profound Reversible Seasonal Changes of Individual Skull Size in a Mammal. Curr Biol, 27(20):3576. DOI 10.1016/j.cub.2017.10.064.

27. Lapoint S, Keicher L, Wikelski M, et al., 2017, Growth Overshoot and Seasonal Size Changes in the Skulls of Two Weasel Species. R Soc Open Sci, 4(1):160947. DOI 10.1098/rsos.160947.

28. Lázaro J, Hertel M, Sherwood CC, et al., 2018, Profound Seasonal Changes in Brain Size and Architecture in the Common Shrew. Brain Struct Funct, 223:2823-40. DOI 10.1007/s00429-018-1666-5.

29. Tallinen T, Chung JY, Rousseau F, et al., 2016, On the Growth and form of Cortical Convolutions. Nat Phys, 12(6):588-93.

30. Checa S, Rausch MK, Petersen A, et al., 2014, The Emergence of Extracellular Matrix Mechanics and Cell Traction Forces as Important Regulators of Cellular Self-organization. Biomech Model Mechanobiol, 14(1):1-13. DOI 10.1007/s10237-014-0581-9.

31. Galbraith CG, Mand YK, Sheetz MP, 2002, The Relationship between Force and Focal Complex Development. J Cell Biol, 159(4):695-705. DOI 10.1083/jcb.200204153.

32. Riveline D, Zamir E, Balaban NQ, et al., 2001, Focal Contacts as Mechanosensors. Externally Applied Local Mechanical Force Induces Growth of Focal Contacts by an Mdia1-Dependent and Rock-Independent Mechanism. J Cell Biol, 153(6):1175-86. DOI 10.1083/jcb.153.6.1175.

33. Corin KA, Gibson LJ, 2010, Cell Contraction Forces in Scaffolds with Varying Pore Size and Cell Density. Biomaterials, 31(18):4835-45. DOI 10.1016/j. biomaterials.2010.01.149.

23. García-Fernández L, Halstenberg S, Unger RE, et al., 2010,
Anti-Angiogenic Activity of Heparin-like Polysulfonated Polymeric Drugs in 3D Human Cell Culture. Biomaterials, 31(31):7863-72. DOI 10.1016/j.biomaterials.2010.07.022.

24. Jand S, Pompe T, 2018, Biomimetic Tumor Microenvironments
Based on Collagen Matrices. Biomater Sci, 6:2009-24. DOI
10.1039/c8bm00303c.

25. Bell E, Ivarsson B, Merrill C, 1979, Production of a Tissuelike
Structure by Contraction of Collagen Lattices by Human Fibroblasts of Different Proliferative Potential in vitro. Proc Natl Acad Sci U S A, 76(3):1274-8. DOI 10.1073/ pnas.76.3.1274.

26. Lázaro J, Dechmann DKN, Lapoint S, et al., 2017, Profound
Reversible Seasonal Changes of Individual Skull Size in a Mammal. Curr Biol, 27(20):3576. DOI 10.1016/j. cub.2017.10.064.

27. Lapoint S, Keicher L, Wikelski M, et al., 2017, Growth Overshoot and Seasonal Size Changes in the Skulls of Two Weasel Species. R Soc Open Sci, 4(1):160947. DOI 10.1098/ rsos.160947.

28. Lázaro J, Hertel M, Sherwood CC, et al., 2018, Profound Seasonal Changes in Brain Size and Architecture in the Common Shrew. Brain Struct Funct, 223:2823-40. DOI 10.1007/s00429-018-1666-5.

29. Tallinen T, Chung JY, Rousseau F, et al., 2016, On the Growth
and form of Cortical Convolutions. Nat Phys, 12(6):588-93.

30. Checa S, Rausch MK, Petersen A, et al., 2014, The Emergence
of Extracellular Matrix Mechanics and Cell Traction Forces as Important Regulators of Cellular Self-organization. Biomech Model Mechanobiol, 14(1):1-13. DOI 10.1007/ s10237-014-0581-9.

31. Galbraith CG, Mand YK, Sheetz MP, 2002, The Relationship
between Force and Focal Complex Development. J Cell Biol, 159(4):695-705. DOI 10.1083/jcb.200204153.

33. Corin KA, Gibson LJ, 2010, Cell Contraction Forces in Scaffolds
with Varying Pore Size and Cell Density. Biomaterials,
31(18):4835-45. DOI 10.1016/j.biomaterials.2010.01.149.

Previous article in this issue

Next article in this issue

International Journal of Bioprinting, Electronic ISSN: 2424-8002 Print ISSN: 2424-7723, Published by AccScience Publishing