Combination effect of p-hydroxybenzyl alcohol and mesenchymal stem cells on the recovery of brain damage in a rat model of brain ischemia

Phatcharida Kaengkan,백승은,최용원,감경윤,김지영,Yan Ru Wu,도병록,강성구 한국통합생물학회 2013 Animal cells and systems Vol.17 No.3

p-Hydroxybenzyl alcohol (HBA), an active component of Gastrodia elata blume, has been reported to provide neuroprotection by preventing brain damage. Transplantation of mesenchymal stem cell (MSC) has been shown to ameliorate ischemic brain injury in animals. To explore a way that might enhance neuroprotection after brain stroke,we investigated whether the transplantation of neural progenitor cells (NPCs) derived from MSCs from adipose tissue combined with HBA may enhance neuroprotective effects in an animal model of brain stroke. Intracarotid injection of combination therapy groups showed a significant reduction of infarct volume by 2,3,5-triphenyltetrazolium chloride staining and an improvement of neurological functions were observed at 3 days after middle cerebral artery occlusion (MCAO), compared to monotherapy groups. In our studies, immunohistochemistry showed that NPCs are more likely to enter a damaged brain than a contralateral nonischemic brain. Coadministration of HBA and NPCs enhanced the anti-apoptotic effect and reduced the number of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling-positive apoptotic cells compared to the vehicle and NPCs at 7 days after MCAO. HBA and the combination of HBANPCs induced the expression of genes encoding antioxidant proteins, including PDI, Nrf2, endogenous neurotrophic factor gene brain-derived neurotrophic factor, NGF, and VEGF, which enhances angiogenesis in an ischemic brain. These effects might be responsible for the survival of NPCs and improved functional behavior. Our finding indicates that combination therapy of HBA and NPCs enhances neuroprotection against ischemic brain injury.

Combination effect of memantine and mesenchymal stem cells on the recovery of brain damage in a rat model of brain ischemia

Phatcharida Kaengkan,감경윤,도병록,강성구 한국통합생물학회 2015 Animal cells and systems Vol.19 No.2

Whether the transplantation of neural progenitor cells (NPCs) derived from mesenchymal stem cells combined with memantine enhanced neuroprotective effects in an animal model of stroke was investigated. In combination therapy groups, significant reduction of infarct volume and improvement of neurological functions were observed at 3 days after middle cerebral artery occlusion (MCAO) compared to monotherapy. Co-administration of memantine and NPCs enhanced the antiapoptotic effect and reduced the number of terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)- positive cells compared to the NPCs alone group at 7 days after MCAO. Intracarotid transplanted NPCs were localized to the ischemic boundary zone. Moreover, the number of transplanted NPCs on day 7 with combination therapy was significantly higher than those of NPCs monotherapy. Memantine and the combination of memantine + NPCs induced the expression of endogenous neurotrophic factor genes brain-derived neurotrophic factor, nerve growth factor, and gliaderived neurotrophic factor and also upregulated Arc (activity-regulated cytoskeleton associated protein). These effects might be responsible for the induced survival of NPCs and improved functional behavior. These finding indicates that combination therapy of memantine and NPCs enhances neuroprotection against ischemic brain injury.

Administration of Mesenchymal Stem Cells and Ziprasidone Enhanced Amelioration of Ischemic Brain Damage in Rats

Phatcharida Kaengkan,강성구,백승은,김지영,감경윤,도병록,이은신 한국분자세포생물학회 2013 Molecules and cells Vol.36 No.6

Ziprasidone is a benzisothiazolyl piperazine derivative that was developed from the chemically related antipsychotic drug tiospirone, and it improves neurological functions of the ischemic brain and is effective in treatment of schi-zophrenia. Mesenchymal stem cells (MSCs) are considered as a leading candidate for neurological regenerative therapy because of their neural differentiation properties in damaged brain. We investigated whether the transplanta-tion of neural progenitor cells (NPCs) derived from adipose mesenchymal stem cells combined with ziprasidone enhances neuroprotective effects in an animal model of focal cerebral ischemia. In combination therapy groups, significant reduction of infarct volume and improvement of neurological functions were observed at 3 days after middle cerebral artery occlusion (MCAO) compared with monotherapy. Co-administration of ziprasidone and NPCs enhanced the anti-apoptotic effect and reduced the num-ber of terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive apoptotic cells compared with the NPCs alone group at 7 days after MCAO. Ziprasi-done or the combination of ziprasidone and NPCs induced the expression of endogenous neurotrophic factor gene brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), and glial cell-derived neurotrophic factor (GDNF). The immu-nohistochemical investigation revealed that the ziprasidone and NPCs attenuated the increased intensity of microglial marker (Iba-1) in the infarcted cortical area. Moreover, the number of transplanted NPCs on day 7 with combination therapy was significantly higher than with NPCs alone. These effects might be responsible for improved functional behavior and increased survival of NPCs. Our finding indicates that combination therapy of ziprasidone and NPCs enhances neuroprotection against ischemic brain injury.

Dopaminergic neural differentiation of human adipose-derived mesenchymal stem cells transplanted in a rat model of Parkinson's disease

A-Reum Kang,Chae Yeon Lee,Kaengkan Phatcharida,Yong-Won Choi,Kyung Yoon Kam,Byung-Rok Do,Sang Hae Kim,Sung Goo Kang 한국발생생물학회 2010 한국발생생물학회 학술발표대회 Vol.29 No.-

Mesenchymal stem cells constitute an potential cellular source to promote brain regeneration with Parkinson's disease. Mesenchymal stem cells have significant advantages over other stem cell types and greater potential for immediate clinical application. The purpose of this study was to investigate whether hMSCs from the human adipose tissue could be induced to differentiate into dopaminergic cells and to assess the developmental potential of hMSC for selectively replacing the midbrain dopamine neurons lost in Parkinson's disease in vitro and in vivo. MSCs were cultured under conditions that promote differentiation of dopaminergic neuron. Using media that include SHH, FGF8, and GDNF. the MSCs were induced in vitro to become dopaminergic neurons. The expressions of the LIM homeobox transcription factor 1, alpha (Lmx1a), tyrosine hydroxylase(TH) proteins were determined by immunofluorescence. Lmx1a has been shown sufficient to confer neurogenic activity on mesencephalic floor plate cells and to determine a mesencephalic dopaminergic neurons fate. This result suggests that hMSCs have the ability to differfentiate into dopaminergic neurons. hMSCs were then transplanted into the striatal in a rat model of Parkinson's disease. The rats were unilaterally lesioned in the substantia nigra with 6-hydroxydopamine and were tested for rotational apomorphine-induced behavior. Following differentiation of dopaminergic neuron, cells displayed dopaminergic morphology and that they expressed dopaminergic marks genes. Finally transplantation of hMSCs into the striatal of Parkinsonian rats resulted in improvement of their behavioral deficits by apomorphine-induced rotational behavior. The hMSCs transplanted rats were proved to be better than compared with the transplantation of PBS. Immunohistochemical analysis of grafted brains revealed that abundant hMSCs survived from the grafts and some of them displayed dopaminergic marks. Our results indicate that hMSC may serve as a good cell source for the treatment of neurodegenerative diseases and have high potential for being used in multiple applications. This cellular approach might become a restorative therapy in Parkinson's disease.

Neuroprotective properties and neural differentiation of the human adipose tissue-derived mesenchymal stem cells transplanted in ischemic rat brain injury

Chae Yeon Lee,A-Reum Kang,Kaengkan Phatcharida,Yong-Won Choi,Kyung Yoon Kam,Byung-Rok Do,Sang Hae Kim,Sung Goo Kang 한국발생생물학회 2010 한국발생생물학회 학술발표대회 Vol.29 No.-

Mesenchymal stem cells (MSCs) has been reported as multipotent progenitor cells that can be expanded rapidly in vitro and differentiated into multiple mesodermal cell type. Human MSCs have been reported to be associated with neural differentiation especially in the cholinergic phenotype in several neural system. In this study, We investigated the ability of MSCs derived human aipose tissue to differentiation into neural cells expressing Islet-1 and further differentiates into cholinergic neurons in cholinergic differentiation media. Immunocytochemistry was performed to detect the expression of Islet-1 and demonstrate characteristic of neurons and cholinergic neurons. Islet-1 was massively detected in the induction stage. Following cholinergic differentiation from Islet-1-expressing MSCs, Cholinergic neuron marker ChAT was higly expressed. Also we examined the neuroprotective effects and neural differentiation of transplanted human adipose tissue-derived mesenchymal stem cells (AT-MSCs) in ischemic stroke. For transplantation, after 3days after MCAO. animal were divided into 2 group: Group A : injected phosphate buffered saline (PBS;5 ㎕ n=10), Group B: transplanted AT-MSCs (5×105 cells, n=10). Each animal received an injection into the right penumbra region (from bregma : AP;-1.3 ㎜, ML;-4.0 ㎜, DV;-5.9 ㎜). In all animals, behavior test were performed at 1, 3, 6, 9, 12, 15 days after MCAO, that was conducted by investigators who were blined to the experimental groups. mNSS test demonstrated that motor, sensory, and balance behavior were impaired after MCAO ischemic insult. Ischemic rats that received AT-MSCs exhibited significantly improved functional performance compared with PBS injected animals and histological analysis revealed that transplanted AT-MSCs expressed marker for neuron. These results suggest that AT-MSCs can be differentiated into neuron especially in cholinergic neuron and may be a potential source of treatment for neurodegenerative disease such as stroke.