Aquapuncture Using Stem Cell Therapy to Treat Mdx Mice

Abstract

Duchenne muscular dystrophy (DMD) occurs due to genetic mutations that lead to absence or decrease of dystrophin protein generating progressive muscle degeneration. Cell therapy using mesenchymal stem cell (MSC) has been described as a treatment to DMD. In this work, MSC derived from deciduous teeth, called stem cells from human exfoliated deciduous teeth (SHED), were injected in acupoint as an alternative therapy to minimize muscle degeneration in twenty-two mdx mice. The treatment occurred three times with intervals of 21 days, and animals were analyzed four times: seven days prior treatment (T-7); 10 days after first treatment (T10); 10 days after second treatment (T31); and 10 days after third treatment (T52). Animals were evaluated by wire test for estimate strength and blood was collected to perform a creatinine phosphokinase analysis. After euthanasia, cranial tibial muscles were collected and submitted to histological and immunohistochemistry analyses. Treated groups presented improvement of strength and reduced creatinine phosphokinase levels. Also, a slight dystrophin increase was observed in tibial cranial muscle when aquapuncture was associated SHED. All therapies have minimized muscle degeneration, but the association of aquapuncture with SHED appears to have better effect, reducing muscle damage, suggesting a therapeutic value.

The Efficacy of Naïve versus Modified Mesenchymal Stem Cells in Improving Muscle Function in Duchenne Muscular Dystrophy: A Systematic Review

“…Only one was non-comparative and was also the only study done on human patients with Duchenne Muscular Dystrophy (Dai et al [ 19 ]). Nine of the studies experimented on x-linked muscular dystrophy (mdx) mice, which are considered equivalent to Duchenne muscular dystrophy, with six using MSCs from humans (Siemionow M et al (2019) [ 20 ], Pang et al [ 21 ], Valadares et al [ 22 ], Esper et al [ 23 ], Siemionow M et al (2021) [ 24 ], and Nitahara-Kashara et al [ 25 ]), three using cells from Sprague-Dawley rats (Geng et al [ 26 ], Li et al [ 27 ], and Nitahara-Kashara et al [ 25 ]), and three studies using cells from utrophin/dystrophin double knockout (dko) mice. Mdx are the most widely used animal model for DMD research overall.…”

“…Two studies investigated cardiac function using echocardiography and/or electromyography (Dai et al [ 19 ], Siemionov et al (2019) [ 20 ]), and one study investigated respiratory function using spirometry (Dai et al). Four studies investigated muscle function using the rotarod performance test as a method (Pang et al [ 21 ], Valadares et al [ 22 ], Geng et al [ 26 ], Li et al [ 27 ]), three studies used electrodes to measure muscle function (Siemionov et al (2021) [ 24 ], Ruehle et al [ 28 ], Rousseau et al [ 29 ]), and two studies used a wire test to measure motor function (Esper et al [ 23 ], Siemionov et al (2021) [ 24 ]). All of the studies also measured other surrogates in conjunction with immunohistochemical and/or histological analysis to confirm muscle function.…”

“…Then, 1 × 10 6 cells (or vehicle) were injected intraperitoneally into the mice once a week for 8 weeks without any immunosuppression. Esper et al [ 23 ] isolated MSCs from human deciduous for pulp digestion. After trypsinization, cells were washed twice and 1 × 10 4 cells were suspended in 20 μL of saline and injected once every three weeks.…”

“…Valadares et al [ 22 ] injected a variety of cell types (fibroblast, myoblast, endometrial pericyte, fallopian tube pericyte, adipose pericyte, and adipose pericyte) into dko mice and used vehicle (HBSS) injection only in dko mice as a control. Esper et al [ 23 ] were interested in the therapeutic effects of aquapuncture and used false aquapuncture with stem cells from human exfoliated deciduous teeth (SHED), true aquapuncture with saline, and true aquapuncture with SHED in mice with no aquapuncture as a control. Rousseau et al [ 29 ] transplanted mdx mice with muscle progenitor cells derived from normal 10 J mice and mdx mice.…”

“…Siemionow et al’s (2019)* [ 20 ] study was the only one to measure cardiac function using echocardiography, and it found significant improvements in ejection fraction and fractional shortening. The remaining seven studies (Pang et al [ 21 ], Esper et al* [ 23 ], Siemionow et al (2021)* [ 24 ], Nitahara-Kashara et al* [ 25 ], Geng et al* [ 26 ], Ruehle et al* [ 28 ], Rousseau et al [ 29 ]) only measured limb function, and all of them were able to find improvements.…”