In this study, we aim to determine the in vivo effect of human umbilical cord blood-derived multipotent stem cells (hUCB-MSCs) on neuropathic pain, using three, principal peripheral neuropathic pain models. Four weeks after hUCB-MSC transplantation, we observed significant antinociceptive effect in ...
In this study, we aim to determine the in vivo effect of human umbilical cord blood-derived multipotent stem cells (hUCB-MSCs) on neuropathic pain, using three, principal peripheral neuropathic pain models. Four weeks after hUCB-MSC transplantation, we observed significant antinociceptive effect in hUCB-MSC-transplanted rats compared to that in the vehicle-treated control. Spinal cord cells positive for c-fos, CGRP, p-ERK, p-p 38, MMP-9 and MMP 2 were significantly decreased in only CCI model of hUCB-MSCs-grafted rats, while spinal cord cells positive for CGRP, p-ERK and MMP-2 significantly decreased in SNL model of hUCB-MSCs-grafted rats and spinal cord cells positive for CGRP and MMP-2 significantly decreased in SNI model of hUCB-MSCs-grafted rats, compared to the control 4 weeks or 8weeks after transplantation (p<0.05). However, cells positive for TIMP-2, an endogenous tissue inhibitor of MMP-2, were significantly increased in SNL and SNI models of hUCB-MSCs-grafted rats. Taken together, subcutaneous injection of hUCB-MSCs may have an antinociceptive effect via modulation of pain signaling during pain signal processing within the nervous system, especially for CCI model. Thus, subcutaneous administration of hUCB-MSCs might be beneficial for improving those patients suffering from neuropathic pain by decreasing neuropathic pain activation factors, while increasing neuropathic pain inhibition factor.
In this study, we aim to determine the in vivo effect of human umbilical cord blood-derived multipotent stem cells (hUCB-MSCs) on neuropathic pain, using three, principal peripheral neuropathic pain models. Four weeks after hUCB-MSC transplantation, we observed significant antinociceptive effect in hUCB-MSC-transplanted rats compared to that in the vehicle-treated control. Spinal cord cells positive for c-fos, CGRP, p-ERK, p-p 38, MMP-9 and MMP 2 were significantly decreased in only CCI model of hUCB-MSCs-grafted rats, while spinal cord cells positive for CGRP, p-ERK and MMP-2 significantly decreased in SNL model of hUCB-MSCs-grafted rats and spinal cord cells positive for CGRP and MMP-2 significantly decreased in SNI model of hUCB-MSCs-grafted rats, compared to the control 4 weeks or 8weeks after transplantation (p<0.05). However, cells positive for TIMP-2, an endogenous tissue inhibitor of MMP-2, were significantly increased in SNL and SNI models of hUCB-MSCs-grafted rats. Taken together, subcutaneous injection of hUCB-MSCs may have an antinociceptive effect via modulation of pain signaling during pain signal processing within the nervous system, especially for CCI model. Thus, subcutaneous administration of hUCB-MSCs might be beneficial for improving those patients suffering from neuropathic pain by decreasing neuropathic pain activation factors, while increasing neuropathic pain inhibition factor.
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제안 방법
In this study, we showed that a single transplantation of hUCB-MSCs reduced mechanical allodynia 1 month after treatment, with a significant difference in peak levels in all three neuropathic pain models studied. This increased mechanical threshold in vivo was confirmed in terms of antibody expression levels.
The aim of this study is to evaluate if subcutaneous transplantation of the hUCB-MSCs can exert antinociceptive effects in vivo, using three different kinds of rat peripheral neuropathic pain models. Regarding pharmacological mechanisms of hUCB-MSCs for its antinociceptive effects in vivo, we studied the antinociceptive effects of the transplanted hUCB-MSCs on biomolecular expression levels of MMP family and TIMP-2, an endogenous tissue inhibitor of MMP-2, and pain-related molecules such as c-fos, calcitonin gene-related peptide (CGRP), phosphorylated extracellular signal-related kinase (p-ERK), and phosphorylated p38 (p-p38).
A series of ten von Frey monofilaments were applied to the entire plantar surface of the hind paw of CCI rats, the 3rd and 4th interdigit of SNL rats, and the external site of lateral plantar surface of the hind paw of SNI rats, which are innervated by the sural nerve [4,24]. The tests determined pain behavior such as paw withdrawal or licking response to three of five repetitive stimuli.
The present study was carried out in principal, rodent models of peripheral neuropathic pain. Using three different models, we circumvented the limitations of each and simultaneously achieved a comparison of their effects on hUCB-MSCs.
대상 데이터
Male Sprague-Dawley rats (Orient Bio Inc., Gyeonggido, Republic of Korea) weighing 150 to 200 g were used in this study, maintained in cages covered with soft bedding, with 3 animals per cage. The animals were divided into 2 groups, where one was subcutaneously administered phosphate buffer solution (PBS; control group, n=6~7), while the other was subcutaneously administered hUCB-MSCs (hUCB-MSC group, n=10~11).
데이터처리
Statistical significance of difference was determined by Duncan’s t-test.
이론/모형
The rats were sacrificed using the carbon dioxide method [22]. All experiments followed the Guidelines on Ethical Standards for Investigation of Experimental Pain in Animals [23]. Additionally, the study protocol was approved by the Institutional Animal Care and Use Committee of Seoul National University.
The rats were allowed free access to food and water in a temperature and humidity-controlled room (20℃, 60%) with a 12/12 h day/ night cycle (7 am/7 pm). The rats were sacrificed using the carbon dioxide method [22]. All experiments followed the Guidelines on Ethical Standards for Investigation of Experimental Pain in Animals [23].
성능/효과
In nerve injury-induced neuropathic pain models in rats and mice, MMP-9 shows a transient upregulation in injured DRG primary sensory neurons and spinal microglia during early-phase neuropathic pain, whereas MMP-2 shows a delayed response in DRG satellite cells and spinal astrocytes during late-phase [21,40]. Our results showed that MMP-2 expression decreased, with TIMP-2 expression simultaneously increasing in the transplanted group. This is in accordance with other studies showing that MMP-2 activity is regulated by its endogenous inhibitor, TIMP-2 [42,43].
1001 in SNI). p-p38 expression in the hUCB-MSCs-treated rats showed a significant decrease in CCI model (p=0.04) while p-p38 expression in the hUCB-MSCs-treated rats was not significant in SNL and SNI models (Fig. 2D, p=0.56 for SNL, p=0.78 for SNI).
후속연구
In conclusion, our current study shows that transplantation of hUCB-MSCs is a novel strategy to relieve neuropathic pain, with potential as an alternative clinical therapy. However, further studies are required to determine clinical feasibility for long-term use to rule out adverse effects due to immune response.
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