Therapeutic Effects of Human Mesenchymal Stem Cells in a Mouse Model of Cerebellar Ataxia with Neuroinflammation.

Cerebellar ataxias (CAs) are neurological diseases characterized by loss of muscle coordination that is a result of damage and inflammation to the cerebellum. Despite considerable efforts in basic and clinical research, most CAs are currently incurable. In this study, we evaluated the therapeutic potential of human mesenchymal stem cells (hMSCs) against CAs associated with neuroinflammation. We observed that hMSC treatment significantly inhibited the symptoms of ataxia in lipopolysaccharide (LPS)-induced inflammatory CA (ICA) mice, which were recently reported as a potential animal model of ICA, through the anti-inflammatory effect of hMSC-derived TNFα-stimulated gene-6 (TSG-6), the protection of Purkinje cells by inhibition of apoptosis, and the modulatory effect for microglial M2 polarization. Thus, our results suggest that hMSC treatment may be an effective therapeutic approach for preventing or improving ataxia symptoms.

Lipopolysaccharide administration for a mouse model of cerebellar ataxia with neuroinflammation.

Most cerebellar ataxias (CAs) are incurable neurological disorders, resulting in a lack of voluntary control by inflamed or damaged cerebellum. Although CA can be either directly or indirectly related to cerebellar inflammation, there is no suitable animal model of CA with neuroinflammation. In this study, we evaluated the utility of an intracerebellar injection of lipopolysaccharide (LPS) to generate an animal model of inflammatory CA. We observed that LPS administration induced the expression of pro-inflammatory molecules following activation of glial cells. In addition, the administration of LPS resulted in apoptotic Purkinje cell death and induced abnormal locomotor activities, such as impaired motor coordination and abnormal hindlimb clasping posture. Our results suggest that intracerebellar LPS administration in experimental animals may be useful for studying the inflammatory component of CA.

Characterization of Mesenchymal Stem Cells Derived from Patients with Cerebellar Ataxia: Downregulation of the Anti-Inflammatory Secretome Profile.

Mesenchymal stem cell (MSC) therapy is a promising alternative approach for the treatment of neurodegenerative diseases, according to its neuroprotective and immunomodulatory potential. Despite numerous clinical trials involving autologous MSCs, their outcomes have often been unsuccessful. Several reports have indicated that MSCs from patients have low capacities in terms of the secretion of neurotrophic or anti-inflammatory factors, which might be associated with cell senescence or disease severity. Therefore, a new strategy to improve their capacities is required for optimal efficacy of autologous MSC therapy. In this study, we compared the secretory potential of MSCs among cerebellar ataxia patients (CA-MSCs) and healthy individuals (H-MSCs). Our results, including secretome analysis findings, revealed that CA-MSCs have lower capacities in terms of proliferation, oxidative stress response, motility, and immunomodulatory functions when compared with H-MSCs. The functional differences were validated in a scratch wound healing assay and neuron-glia co-cultures. In addition, the neuroprotective and immunoregulatory protein follistatin-like 1 (FSTL1) was identified as one of the downregulated proteins in the CA-MSC secretome, with suppressive effects on proinflammatory microglial activation. Our study findings suggest that targeting aspects of the downregulated anti-inflammatory secretome, such as FSTL1, might improve the efficacy of autologous MSC therapy for CA.

No Synergistic Effect of Silibinin and Morin in a Kainic Acid-Induced Epileptic Mouse Model.

Temporal lobe epilepsy (TLE) is the most common form of localization-related epilepsy, with the highest prevalence rate in adulthood. Recently, we reported the beneficial effects of the individual treatment with flavonoids such as silibinin and morin in kainic acid (KA)-treated mouse model for TLE. In this study, we investigated whether there is a synergistic effect of co-treatment with silibinin and morin on the susceptibility to seizure, the frequency of spontaneous recurrent seizures (SRSs), and granule cell dispersion in the dentate gyrus, which could be partially controlled by treatment with each flavonoid in the animal model for TLE. Unfortunately, we did not observe any synergistic effect against the susceptibility of seizure and SRS induced by KA treatment. However, the combination of these flavonoids showed similar antiepileptic effects compared with treatment with each one individually. Therefore, although silibinin and morin are not suitable for combination therapy, our results still suggest that these flavonoids can be used as potent therapeutic compounds for preventing epileptic seizures.

Beneficial Effects of Hesperetin in a Mouse Model of Temporal Lobe Epilepsy.

Abnormal reorganization of the dentate gyrus and neuroinflammation in the hippocampus represent characteristic phenotypes of patients suffering from temporal lobe epilepsy. Hesperetin, a flavanone abundant in citrus fruit, is known to have protective effects by preventing inflammation and oxidative stress in neuronal cultures and in the adult murine brain. However, the protective effects of hesperetin against epileptic seizures in vivo remain unclear, despite one study reporting anticonvulsant effects in vitro. In this study, we report that oral administration of hesperetin not only delays the onset of seizures triggered by kainic acid (KA) but also contributes to the attenuation of granule cell dispersion in the KA-treated hippocampus. Moreover, we observed that hesperetin administration inhibited the expression of pro-inflammatory molecules produced by activated microglia in the hippocampus. Thus, administration of hesperetin might be beneficial for preventing epileptic seizures.

Morin Prevents Granule Cell Dispersion and Neurotoxicity via Suppression of mTORC1 in a Kainic Acid-induced Seizure Model.

An abnormal reorganization of the dentate gyrus and neurotoxic events are important phenotypes in the hippocampus of patients with temporal lobe epilepsy (TLE). The effects of morin, a bioflavonoid constituent of many herbs and fruits, on epileptic seizures have not yet been elucidated, though its beneficial effects, such as its anti-inflammatory and neuroprotective properties, are well-described in various neurodegenerative diseases. In the present study, we investigated whether treatment with morin hydrate (MH) can reduce the susceptibility to seizures, granule cell dispersion (GCD), mammalian target of rapamycin complex 1 (mTORC1) activity, and the increases in the levels of apoptotic molecules and inflammatory cytokines in the kainic acid (KA)-induced seizure mouse model. Our results showed that oral administration of MH could reduce susceptibility to seizures and lead to the inhibition of GCD and mTORC1 activity in the KA-treated hippocampus. Moreover, treatment with MH significantly reduced the increased levels of apoptotic signaling molecules and pro-inflammatory mediators in the KA-treated hippocampus compared with control mice, suggesting a neuroprotective role. Therefore, these results suggest that morin has a therapeutic potential against epilepsy through its abilities to inhibit GCD and neurotoxic events in the in vivo hippocampus.

Beneficial Effects of Silibinin Against Kainic Acid-induced Neurotoxicity in the Hippocampus in vivo.

Silibinin, an active constituent of silymarin extracted from milk thistle, has been previously reported to confer protection to the adult brain against neurodegeneration. However, its effects against epileptic seizures have not been examined yet. In order to investigate the effects of silibinin against epileptic seizures, we used a relevant mouse model in which seizures are manifested as status epilepticus, induced by kainic acid (KA) treatment. Silibinin was injected intraperitoneally, starting 1 day before an intrahippocampal KA injection and continued daily until analysis of each experiment. Our results indicated that silibinin-treatment could reduce seizure susceptibility and frequency of spontaneous recurrent seizures (SRS) induced by KA administration, and attenuate granule cell dispersion (GCD), a morphological alteration characteristic of the dentate gyrus (DG) in temporal lobe epilepsy (TLE). Moreover, its treatment significantly reduced the aberrant levels of apoptotic, autophagic and pro-inflammatory molecules induced by KA administration, resulting in neuroprotection in the hippocampus. Thus, these results suggest that silibinin may be a beneficial natural compound for preventing epileptic events.

Endogenous inspired biomineral-installed hyaluronan nanoparticles as pH-responsive carrier of methotrexate for rheumatoid arthritis.

Methotrexate (MTX), an anchor drug for rheumatoid arthritis (RA), has been suffered from refractoriness and high toxicity limiting effective dosage. To mitigate these challenges, the ability to selectively deliver MTX to arthritis tissue is a much sought-after modality for the treatment of RA. In this study, we prepared mineralized nanoparticles (MP-HANPs), composed of PEGylated hyaluronic acid (P-HA) as the hydrophilic shell, 5ß-cholanic acid as the hydrophobic core, and calcium phosphate (CaP) as the pH-responsive mineral. Owing to the presence of CaP as the diffusion barrier, mineralized HANPs revealed the pH-responsiveness of release kinetics of MTX across neutral to acidic conditions. HANPs were internalized via receptor-mediated endocytosis in macrophages which involved molecular redundancy among major hyaladherins, including CD44, stabilin-2, and RHAMM. Following endocytosis, MP-HANPs loaded with doxorubicin revealed pH-dependent demineralization followed by dramatic acceleration of drug release into the cytosol compared to other HANPs. Furthermore, an in vivo study showed a significantly high paw-to-liver ratio of fluorescent intensity after systemic administration of MP-HANP-Cy5.5, indicating improved biodistribution of nanoparticles into arthritic paws in collagen-induced arthritis mice. Treatment with MTX-loaded MP-HANPs ameliorated inflammatory arthritis with remarkable safety at high dose of MTX. We highlight the distinct advantages of combining key benefits of biomineralization and PEGylation with HA-based nanoparticles for arthritis-selective targeting, thus suggesting MP-HANPs as a promising carrier of MTX for treatment of RA.

Usage report of pharmacopuncture in musculoskeletal patients visiting Korean medicine hospitals and clinics in Korea.

BACKGROUND: Pharmacopuncture is a relatively new acupuncture therapy combining acupuncture with herbal medicine. While pharmacopuncture is applied extensively in Korean medicine treatment, there are no clinical reports regarding what types of pharmacopuncture are used for which diseases. METHODS: Data was extracted retrospectively from the electronic medical records of all inpatients and outpatients at 12 Korean medicine hospitals and clinics during the period of December 17, 2010 to October 2, 2014. Treatment patterns for acupuncture, electroacupuncture and pharmacopuncture were analyzed. Principle diagnosis codes, frequency of treatment, pharmacopuncture type and costs were investigated to assess pharmacopuncture use in clinical settings. RESULTS: During the study period, a total 33,415 inpatients and 373,755 outpatients visited the study sites, and most were musculoskeletal. Among inpatients and outpatients, 98.6 % and 77.6 % received pharmacopuncture, respectively. Administration rate of pharmacopuncture for the 10 most frequent principle diagnosis codes was 97.2-99.3 % in inpatients, and that for outpatients was 73.0-91.5 %. The average number of pharmacopuncture sessions in pharmacopuncture recipients was 8.2 ± 12.3 for outpatients and 25.8 ± 18.7 for inpatients. The mean total cost for pharmacopuncture per patient was $556.24 ± 174.62 among inpatients, and $149.16 ± 243.85 among outpatients. Estimated average cost per pharmacopuncture session was $23-24 for inpatients, and $17-18 for outpatients. Shinbaro1, bee venom, Hwangryunhaedok, and Shinbaro2 were the most frequently used pharmacopuncture types. CONCLUSIONS: This is the first analysis of treatment patterns of pharmacopuncture in a large-scale Korean medicine hospital/clinic patient population. We verified patterns of pharmacopuncture use for musculoskeletal disease treatment in Korea, and use of pharmacopuncture varied depending on disease or symptom severity. These results are expected to contribute to future clinical study design and standardization of pharmacopuncture.

Volume-normalized transfer constant as an imaging biomarker for chronic inflammatory arthritis: A dynamic contrast enhanced MRI study.

PURPOSE: Proper diagnosis and treatment of rheumatoid arthritis are extremely important to optimize treatment outcomes. Dynamic contrast-enhanced MRI (DCE-MRI) may be used as a biomarker to detect inflammatory changes in synovial joints and to discriminate active and inactive stages of the disease. The aim of this study was to investigate vascular permeability changes associated with inflammatory arthritis progression and its treatment. METHODS: Arthritis was induced in DBA/1J mice by immunization with type-II collagen emulsified in complete Freund's adjuvant. Severity of arthritis was monitored using the clinical arthritis index. MR images of mice were obtained at different stages of arthritis progression and at 3 weeks after methotrexate treatment. Immunohistochemical staining using an anti-CD31 antibody was used to assess vessel density. RESULTS: Volume-normalized transfer constant increased progressively until the active stage of arthritis was reached, and thereafter declined gradually. The pattern of volume-normalized transfer constant changes quantified using DCE-MRI correlated with vascular densities and immunohistochemical findings. Furthermore, volume-normalized transfer constant and densities decreased significantly in a dose-dependent manner after treatment with methotrexate. CONCLUSION: Volume-normalized transfer constant assessed by DCE-MRI can be used as an imaging biomarker for tracking disease progression and for monitoring therapeutic efficacy in inflammatory arthritis. Magn Reson Med 76:926-934, 2016. © 2016 Wiley Periodicals, Inc.