Collagen/silk fibroin scaffold combined with human umbilical cord-derived mesenchymal stem cells in the treatment of traumatic brain injury in dogs

doi:10.12307/2023.725

Abstract

Abstract: BACKGROUND: So far, the subversive treatment of traumatic brain injury is very limited. The combination of medicine and engineering has brought new prospects for the repair and regeneration of central nervous system nerves.
OBJECTIVE: To investigate the therapeutic effect of collagen/silk fibroin scaffold that can carry seed cells and is safe and porous, combined with human umbilical cord-derived mesenchymal stem cells on traumatic brain injury in beagle dogs.
METHODS: (1) Passage 3 human umbilical cord-derived mesenchymal stem cells were inoculated onto collagen/silk fibroin scaffold. The growth of human umbilical cord-derived mesenchymal stem cells was observed by inverted phase contrast microscope, scanning electron microscope, immunofluorescence staining and hematoxylin-eosin staining. (2) Twenty-four beagle dogs were randomly divided into four groups. In the trauma group, only the traumatic brain injury model was established without other treatment. In the stem cell group, human umbilical cord-derived mesenchymal stem cells were transplanted after model establishment. In the scaffold group, the collagen/silk fibroin scaffold was transplanted after model establishment. In the combination group, human umbilical cord-derived mesenchymal stem cells and collagen/silk fibroin scaffold were transplanted after model establishment. The grafts were all transplanted locally in the injured area. At 1 day, 1, 4, 8, 12, 16, 20 and 24 weeks after operation, a modified Glasgow Coma Scale was performed respectively. Motor-evoked potentials were detected at 1, 3 and 6 months after operation. At 6 months after operation, magnetic resonance imaging and in situ hybridization of brain tissue repair RNA were performed to evaluate the recovery of traumatic brain injury.
RESULTS AND CONCLUSION: (1) Human umbilical cord-derived mesenchymal stem cells attached to collagen/silk fibroin scaffold surface grew well and extended many pseudopods. (2) Modified Glasgow Coma Scale score of the combination group was significantly higher than that of the trauma group, stem cell group and scaffold group at 1, 4, 8, 12, 16, 20 and 24 weeks after operation (P < 0.05). (3) The latency and amplitude of motor-evoked potential of the combination group at 1, 3 and 6 months were significantly better than those of the trauma group under different constant pressure stimuli (P < 0.01). (4) Diffusion tensor imaging of magnetic resonance scanning showed that the integrity of the corticospinal tract in the combination group was better than that in the trauma, stem cell and scaffold groups, and a new corticospinal tract could be seen on the injured side. (5) The results of in situ hybridization of brain tissue repair RNA demonstrated that the expression levels of synaptophysin, microtubule-associated protein 2, von Willebrand factor, neurofilament protein and MBP in the combination group were more than those in the trauma, stem cell and scaffold groups. (6) It is indicated that human umbilical cord-derived mesenchymal stem cells combined with collagen/silk fibroin scaffold play an active role in corticospinal tract regeneration, motor function recovery, angiogenesis and neurite growth in dogs with traumatic brain injury.

Key words: biomedical engineering, collagen/silk fibroin, human umbilical cord-derived mesenchymal stem cell, traumatic brain injury, beagle dog

CLC Number:

Wang Ziqi, Li Xiaoyin, Jiang Jipeng, Song Zhen, Li Zhengchao, Chen Shulian, Chen Xuyi. Collagen/silk fibroin scaffold combined with human umbilical cord-derived mesenchymal stem cells in the treatment of traumatic brain injury in dogs[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(34): 5483-5490.

Figures/Tables 11

References

[1] MENON DK, SCHWAB K, WRIGHT DW, et al. Position statement: definition of traumatic brain injury. Arch Phys Med Rehabil. 2010;91(11):1637-1640.
[2] GBD 2016 NEUROLOGY COLLABORATORS. Global, regional, and national burden of neurological disorders, 1990-2016: a systematic analysis for the Global Burden of Disease Study 2016. Lancet Neurol. 2019;18(5):459-480.
[3] MAAS AIR, FITZGERALD M, GAO G, et al. Traumatic brain injury over the past 20 years: research and clinical progress. Lancet Neurol. 2022;21(9):768-770.
[4] JIANG JY, GAO GY, FENG JF, et al. Traumatic brain injury in China. Lancet Neurol. 2019;18(3):286-295.
[5] ANCANS J. Cell therapy medicinal product regulatory framework in Europe and its application for MSC-based therapy development. Front Immunol. 2012;3:253.
[6] HOANG DM, PHAM PT, BACH TQ, et al. Stem cell-based therapy for human diseases. Signal Transduct Target Ther. 2022;7(1):272.
[7] MAAS AIR, MENON DK, ADELSON PD, et al. Traumatic brain injury: integrated approaches to improve prevention, clinical care, and research. Lancet Neurol. 2017;16(12):987-1048.
[8] DEWAN MC, RATTANI A, GUPTA S, et al. Estimating the global incidence of traumatic brain injury. J Neurosurg. 2019;130(4):1080-1097.
[9] ISMAIL H, SHAKKOUR Z, TABET M, et al. Traumatic Brain Injury: Oxidative Stress and Novel Anti-Oxidants Such as Mitoquinone and Edaravone. Antioxidants (Basel). 2020;9(10):943.
[10] NAKAO M, INANAGA D, NAGASE K, et al. Characteristic differences of cell sheets composed of mesenchymal stem cells with different tissue origins. Regen Ther. 2019;11:34-40.
[11] ZHAO J, YU G, CAI M, et al. Bibliometric analysis of global scientific activity on umbilical cord mesenchymal stem cells: a swiftly expanding and shifting focus. Stem Cell Res Ther. 2018;9(1):32.
[12] EL OMAR R, BEROUD J, STOLTZ JF, et al. Umbilical cord mesenchymal stem cells: the new gold standard for mesenchymal stem cell-based therapies? Tissue Eng Part B Rev. 2014;20(5):523-544.
[13] SUN W, GREGORY DA, TOMEH MA, et al. Silk Fibroin as a Functional Biomaterial for Tissue Engineering. Int J Mol Sci. 2021;22(3):1499.
[14] BUITRAGO JO, PATEL KD, EL-FIQI A, et al. Silk fibroin/collagen protein hybrid cell-encapsulating hydrogels with tunable gelation and improved physical and biological properties. Acta Biomater. 2018;69:218-233.
[15] TU Y, CHEN C, SUN HT, et al. Combination of temperature-sensitive stem cells and mild hypothermia: a new potential therapy for severe traumatic brain injury. J Neurotrauma. 2012;29(14):2393-2403.
[16] LIU X, ZHANG G, WEI P, et al. Three-dimensional-printed collagen/chitosan/secretome derived from HUCMSCs scaffolds for efficient neural network reconstruction in canines with traumatic brain injury. Regen Biomater. 2022; 9:rbac043.
[17] JIANG J, DAI C, NIU X, et al. Establishment of a precise novel brain trauma model in a large animal based on injury of the cerebral motor cortex. J Neurosci Methods. 2018;307:95-105.
[18] CAMERON S, WELTMAN JG, FLETCHER DJ. The prognostic value of admission point-of-care testing and modified Glasgow Coma Scale score in dogs and cats with traumatic brain injuries (2007-2010): 212 cases. J Vet Emerg Crit Care (San Antonio). 2022;32(1):75-82.
[19] ZHU W, CHEN L, WU Z, et al. Bioorthogonal DOPA-NGF activated tissue engineering microunits for recovery from traumatic brain injury by microenvironment regulation. Acta Biomater. 2022;150:67-82.
[20] HSU RS, LI SJ, FANG JH, et al. Wireless charging-mediated angiogenesis and nerve repair by adaptable microporous hydrogels from conductive building blocks. Nat Commun. 2022;13(1):5172.
[21] SUN D, LIU K, LI Y, et al. Intrinsically Bioactive Manganese–Eumelanin Nanocomposites Mediated Antioxidation and Anti‐Neuroinflammation for Targeted Theranostics of Traumatic Brain Injury. Adv Healthc Mater. 2022; 11(16):e2200517.
[22] WEIGHTMAN AP, PICKARD MR, YANG Y, et al. An in vitro spinal cord injury model to screen neuroregenerative materials. Biomaterials. 2014;35(12): 3756-3765.
[23] CHAU AL, GETTY PT, RHODE AR, et al. Superlubricity of pH-responsive hydrogels in extreme environments. Front Chem. 2022;10:891519.
[24] JIANG S, YU Z, ZHANG L, et al. Effects of different aperture-sized type I collagen/silk fibroin scaffolds on the proliferation and differentiation of human dental pulp cells. Regen Biomater. 2021;8(4):rbab028.
[25] MARTÍN-MARTÍN Y, FERNÁNDEZ-GARCÍA L, SANCHEZ-REBATO MH, et al. Evaluation of Neurosecretome from Mesenchymal Stem Cells Encapsulated in Silk Fibroin Hydrogels. Sci Rep. 2019;9(1):8801.
[26] SHAH R, STODULKA P, SKOPALOVA K, et al. Dual Crosslinked Collagen/Chitosan Film for Potential Biomedical Applications. Polymers (Basel). 2019; 11(12):2094.
[27] JIANG X, WU S, KUSS M, et al. 3D printing of multilayered scaffolds for rotator cuff tendon regeneration. Bioact Mater. 2020;5(3):636-643.
[28] MEBARKI M, ABADIE C, LARGHERO J, et al. Human umbilical cord-derived mesenchymal stem/stromal cells: a promising candidate for the development of advanced therapy medicinal products. Stem Cell Res Ther. 2021;12(1):152.
[29] LI J, XU SQ, ZHAO YM, et al. Comparison of the biological characteristics of human mesenchymal stem cells derived from exfoliated deciduous teeth, bone marrow, gingival tissue, and umbilical cord. Mol Med Rep. 2018;18(6):4969-4977.
[30] XIE Q, LIU R, JIANG J, et al. What is the impact of human umbilical cord mesenchymal stem cell transplantation on clinical treatment? Stem Cell Res Ther. 2020;11(1):519.
[31] SUN L, WANG F, CHEN H, et al. Co-Transplantation of Human Umbilical Cord Mesenchymal Stem Cells and Human Neural Stem Cells Improves the Outcome in Rats with Spinal Cord Injury. Cell Transplant. 2019;28(7):893-906.
[32] CAO T, CHEN H, HUANG W, et al. hUC-MSC-mediated recovery of subacute spinal cord injury through enhancing the pivotal subunits β3 and γ2 of the GABAA receptor. Theranostics. 2022;12(7):3057-3078.
[33] CHEN C, HU N, WANG J, et al. Umbilical cord mesenchymal stem cells promote neurological repair after traumatic brain injury through regulating Treg/Th17 balance. Brain Res. 2022;1775:147711.
[34] CUI L, LUO W, JIANG W, et al. Human umbilical cord mesenchymal stem cell-derived exosomes promote neurological function recovery in rat after traumatic brain injury by inhibiting the activation of microglia and astrocyte. Regen Ther. 2022;21:282-287.
[35] OSIER N, DIXON CE. The Controlled Cortical Impact Model of Experimental Brain Trauma: Overview, Research Applications, and Protocol. Methods Mol Biol. 2016;1462:177-192.
[36] JIANG J, LIU X, CHEN H, et al. 3D printing collagen/heparin sulfate scaffolds boost neural network reconstruction and motor function recovery after traumatic brain injury in canine. Biomater Sci. 2020;8(22):6362-6374.
[37] OLSEN A, BABIKIAN T, DENNIS EL, et al. Functional Brain Hyperactivations Are Linked to an Electrophysiological Measure of Slow Interhemispheric Transfer Time after Pediatric Moderate/Severe Traumatic Brain Injury. J Neurotrauma. 2020;37(2):397-409.
[38] SHARMA D, HOLOWAYCHUK MK. Retrospective evaluation of prognostic indicators in dogs with head trauma: 72 cases (January-March 2011). J Vet Emerg Crit Care (San Antonio). 2015;25(5):631-639.
[39] SAENUBOL P, AKATVIPAT A, PLEUMSAMRAN A, et al. Correlation between bispectral index value and modified Glasgow Coma Scale score in dogs with altered level of consciousness. J Vet Emerg Crit Care (San Antonio). 2021;31(1):52-58.
[40] JANG SH, LEE SJ. Corticoreticular Tract in the Human Brain: A Mini Review. Front Neurol. 2019;10:1188.
[41] LI D, JIAO YM, WANG L, et al. Surgical outcome of motor deficits and neurological status in brainstem cavernous malformations based on preoperative diffusion tensor imaging: a prospective randomized clinical trial. J Neurosurg. 2018;130(1):286-301.
[42] JANG SH, KIM SH, KWON YH. Extensive traumatic axonal injury of brain due to violence: A case report. Medicine (Baltimore). 2018;97(48): e13315.
[43] DOUGLAS DB, RO T, TOFFOLI T, et al. Neuroimaging of Traumatic Brain Injury. Med Sci (Basel). 2018;7(1):2.
[44] LACALLE-AURIOLES M, CASSEL DE CAMPS C, ZORCA CE, et al. Applying hiPSCs and Biomaterials Towards an Understanding and Treatment of Traumatic Brain Injury. Front Cell Neurosci. 2020;14:594304.
[45] LV B, ZHANG X, YUAN J, et al. Biomaterial-supported MSC transplantation enhances cell-cell communication for spinal cord injury. Stem Cell Res Ther. 2021;12(1):36.
[46] MONTICELLI S, NATOLI G. Short-term memory of danger signals and environmental stimuli in immune cells. Nat Immunol. 2013;14(8): 777-784.
[47] ZHU D, JIANG Z, LI N, et al. Insights into the use of genetically modified decellularized biomaterials for tissue engineering and regenerative medicine. Adv Drug Deliv Rev. 2022;188:114413.