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Brain & Heart Human DPSCs attenuated amyotrophic lateral sclerosis in mice

1. Introduction teeth, a bone chisel was used to segment the tooth tissue.
The pulp tissue was extracted and rinsed repeatedly,
Amyotrophic lateral sclerosis (ALS) is a progressive, after which it was cut with curved scissors into small
debilitating disorder that is characterized by the pieces of approximately 0.5 mm . Subsequently, 0.3%
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deterioration of motor neurons that results in muscle collagenase type Ⅰ (MP Biomedicals LLC, CA, USA) and
weakness and atrophy, bulbar palsy, and pyramidal tract 0.4% dispase II (Sigma, SLZ, USA) were added to digest
signs. The incidence of ALS is between 0.6 and 3.8/100,000 for 1 h at 37°C. Digestion was terminated with Dulbecco’s
1,2
person-years and increases with age. This devastating phosphate-buffered saline (PBS), and the cells were
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neurodegenerative disease has a poor prognosis, with 50% inoculated into a six-well cell culture plate and cultured
of patients passing away within 30 months of symptom in a cell incubator at 37°C and 5% CO . Then, the cells
onset and only 20% surviving for 5 – 10 years. Hence, 2
5
the primary aim of current treatment was to extend life were washed and resuspended with 0.9% sterile saline
expectancy in patients with ALS. solution for counting. Two batches of cells were used in the
experiment, and cells were identified by flow cytometry.
The pathophysiology of ALS is the result of complex The hDPSCs were positive for the surface antigens
interactions between genetic and molecular pathways. CD73/CD90/CD44/CD166/CD105/CD146 and negative
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Mutations in the copper-zinc superoxide dismutase 1 for CD11b/CD19/CD34/CD45/HLA-DR (Figure S1).
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(SOD1) gene may be a major factor, because they cause Next, 1 × 10 cells were resuspended in 200 µl of 0.9%
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conformational instability and misfolding of the SOD1 sterile physiological saline and injected into SOD1-G93A
peptide, leading to the formation of intracellular aggregates mice through the tail vein. The SOD1-G93A vehicle group
and disruption of axonal transport systems and cellular and wild-type (WT) mice group were administered 200 µL
functions. At present, no definitive cure exists for ALS. of 0.9% sterile physiological saline. The hDPSCs or 0.9%
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Riluzole and edaravone are the only approved medications sterile physiological saline were infused once through the
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with limited beneficial effects. Nonetheless, there has tail vein at disease onset.
been increasing research in recent years on the use of
stem cells for treating neurodegenerative diseases. 10-12 This 2.2. Mice
therapy works by replacing cells and secreting neurotrophic High-copy SOD1-G93A 72 transgenic mice
factors, which help regenerate neurons and glial cells. 10,11 (B6.Cg-Tg[SOD1∗G93A]1Gur/J) were obtained from
Mesenchymal stem cells (MSCs) originate from the the Jackson Laboratory (Strain#: 004435). PCR genotype
mesoderm during early development and can differentiate identification was performed on mice according to the
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into neurons under certain stimulation conditions. MSCs genotype identification scheme provided by the Jackson
may be a promising potential therapy for ALS treatment. Laboratory. Mice containing transgenic bands (molecular
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Human dental pulp stem cells (hDPSCs) are a type of MSCs size, 236 bp) were identified as SOD1-G93A mice and those
found in dental pulp tissue and possess strong self-renewal without magnetic bands were WT controls. Hemizygous
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and multidirectional differentiation potential. Compared carriers (males) were bred with C57BL/6J inbred mice. The
with other MSCs, hDPSCs are easier to obtain, have SOD1-G93A transgenic mice were obtained by genotyping.
higher proliferation rates, and can also differentiate into All mice were housed in a specific pathogen-free environment
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a variety of somatic cells in an appropriate environment. 16,17 with a 12 – 12-h dark–light cycle and had free access to food
Furthermore, hDPSCs have a lower potential to transform and water. The animals were randomly assigned to individual
into tumors. Therefore, this study was conducted to groups. The endpoint in this study was based on the criteria
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investigate the efficacy of hDPSCs treatment based on
clinical and pathological changes in an ALS mouse model. previously reported by the ALS-Therapy Development
Institute (ALS-TDI), that is, the loss of self-righting ability
2. Methods within 10 s (neurological score = 4) or the inability to move to
reach food on the cage floor. Mice that reached the humane
2.1. Administration of hDPSCs endpoint were euthanized within 3 h. No animal deaths

Here, hDPSCs were extracted from the third molars of occurred during the evaluation, and there were no post-
healthy volunteers at the Dental Clinic of the Beijing infusion deaths. All experimental procedures were conducted
Stomatological Hospital, with ethical approval (Approval according to the guidelines of the Institutional Animal Care
Number: KJ-2021- 016-C-01-CS, Beijing, China). and Use Committee of Beijing Tiantan Hospital and the
Written informed consent was provided to all donors Chinese Small Animal Protection Association Experimental
who participated in the study. The hDPSCs injection was Protocol with approval from the Institutional Animal Care
prepared and appraised by Beijing Chengnuo Medical and Use Committee Institutional Animal Care and Use
Technology Co., Ltd. After cleaning and disinfecting the Committee (IACUC) under the protocol number 202203004.

Volume 2 Issue 4 (2024) 2 doi: 10.36922/bh.3996