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Scooter ! Fabulous reference! The use of opioids is commonly associated with opioid tolerance OT and opioid-induced hyperalgesia OIH , which limit efficacy and compromise safety. We hypothesized that mesenchymal stem cells MSCs attenuate OT and OIH in rats and mice based on the understanding that MSCs possess remarkable anti-inflammatory properties and that both OT and chronic pain are associated with neuroinflammation in the spinal cord.
Remarkably, established OT and OIH were significantly reversed by either intravenous or intrathecal MSCs when cells were transplanted after repeated morphine injections.
The animals did not show any abnormality in vital organs or functions. Immunohistochemistry revealed that the treatments significantly reduced activation level of microglia and astrocytes in the spinal cord.
We have thus demonstrated that MSC-TP promises to be a potentially safe and effective way to prevent and reverse two of the major problems of opioid therapy.
Introduction Chronic pain is a significant public health problem. Opioids, such as morphine, play an indispensable role in pain relief but are often associated with two major problems: opioid tolerance OT and opioid-induced hyperalgesia OIH 3 , 4 , 5 , 6 , 7.
OT is a physiological process where the body adjusts to a medication of frequent exposure and requires escalating doses to achieve the same effect. OIH is a phenomenon, in which individuals taking opioids to treat pain paradoxically develop an increased sensitivity to noxious stimuli. These facts underscore an urgent need for finding effective therapies to treat pain and OT and to reduce the disastrous outcomes associated with opioid treatment.
Distinct molecular mechanisms are indicated for the two closely related but different phenomena 9.
Neuroinflammation, mediated by immune cells and glial cells, appears to play a central role 10 , Opioids such as morphine can cause neuroinflammation 12 through acting on Toll-like receptor 4 on microglia and lead to development of OT We chose to use MSCs because of their powerful paracrine functions, as shown in animal models of diseases such as traumatic brain injury 15 , peripheral neuropathy 16 , 17 , and neuropathic pain Immunomodulatory and anti-inflammatory effects of MSCs were related to neuroprotection, neuroregeneration, and neuroneuromodulation in these studies.
For example, intravenous IV injection of human adipose-derived MSCs hAD-MSCs induced a significant reduction in mechanical allodynia and complete reversal of thermal hyperalgesia in a dose-dependent fashion in a mouse model of diabetic neuropathy We tested these hypotheses by using intrathecal and intravenous routes of transplantation in rats and mice and studied the distribution of the transplanted cells and the level of activation of microglia and astrocytes in the spinal cord in response to morphine and MSC-TP.
These cells showed morphological properties and cell markers characteristic of stem cells and differentiated into osteoblast cells and adipose cells in specific culture media Fig. Figure 1: Isolation and characterization of MSCs from the rat bone marrow. Unstained controls are indicated as blue.
These data represent three individual experiments. Full size image We then tested the preventive and therapeutic effects of intrathecal and intravenous MSC-TPs on OT, which was induced by daily morphine injections. Acute OT was induced after 3 days of daily injections. Chronic OT was induced by daily morphine injections for 3 to 4 weeks and evaluated by measuring two sets of paw withdrawal thresholds PWTs to mechanical and thermal stimulation.
The first were measured before daily morphine injection and the second were measured 50 min after the injection. The differences between the two sets reflect the level of tolerance. A large difference indicates no or low tolerance while a small difference indicates high tolerance.
The differences decreased gradually and significantly after 7 days of morphine treatment, reached a minimal difference at day 12, and maintained a small difference thereafter Supplementary Fig.
OT was further evaluated by the tail flick test. In contrast, it significantly and consistently attenuated the development of OT.
Both intrathecal and intravenous MSCs were remarkably effective Fig. A one-time transplantation significantly mitigated OT for the whole course of the experiments of up to 26 days Fig. The effects were almost identical when the transplantation was performed one day or 7 days before morphine treatment. These experiments were repeated separately by two groups of experimenters who were blinded to the treatments Supplementary Fig.
The effects were further evaluated by the tail flick tests Fig. Pain-like behaviors in rats were assessed by von Frey filament b,c and tail flick tests d,e. OT was induced by repeated daily MS injections at 7. Pain-like behavior was evaluated by von Frey filament in rats f and tail flick tests in rats g and mice h. IT: intrathecal; IV: intravenous. MS, morphine sulfate; PWT, paw withdrawal threshold. Morphine-induced OT reached its peak at day 12 of daily morphine injections in both the mechanical and thermal tests Fig.
Both intrathecal and intravenous MSCs significantly and consistently restored the sensitivity to morphine. This therapeutic effect took place rather rapidly and lasted for the whole course of the experiment to day 28 with no sign of waning. The therapeutic effects were further tested in mice.
Compared to the control group, the mean tail flick latencies were significantly increased in the transplantation group Fig. Consistent with the rat experiments, this effect took place within 2 days of the transplantation.
Next, we tested the preventive and therapeutic effects of MSCs on OIH, which was also induced by daily morphine injections. OIH was reflected by the progressive decline of PWTs from the baseline values established 3 days before morphine treatment Supplementary Fig.
The effects were long-lasting with no sign of waning over time. Consistent results were observed whether the transplantation was performed one day or seven days before morphine treatment.
These results were replicated by two groups of experimenter who were blinded to the treatments. The transplantation effectively and rapidly reversed OIH. This effect lasted for the whole duration of the experiments Fig.
The therapeutic effect was further tested in mice Fig. OIH was induced by daily MS injections of 7. Pain-like behavior was assessed by von Frey filament c or tail flick tests d. MSC-TP of both routes consistently and significantly increased the mean PWT or the mean tail flick latency, indicating reversal of mechanical allodynia and thermal hyperalgesia in both rats and mice.
Full size image All animals survived the entire course of the experiments up to 68 days and had normal locomotion, food and fluid intake, body weight gain, and biochemical parameters for liver and kidney functions Supplementary Fig.