Autophagy induction in the skeletal myogenic differentiation of human tonsil-derived mesenchymal stem cells

Park, Saeyoung,Choi, Yoonyoung,Jung, Namhee,Kim, Jieun,Oh, Seiyoon,Yu, Yeonsil,Ahn, Jung-Hyuck,Jo, Inho,Choi, Byung-Ok,Jung, Sung-Chul D.A. Spandidos 2017 International journal of molecular medicine Vol.39 No.4

<P>Mesenchymal stem cells (MSCs) are capable of self-renewal and differentiation and are thus a valuable source for the replacement of diseased or damaged organs. Previously, we reported that the tonsils can be an excellent reservoir of MSCs for the regeneration of skeletal muscle (SKM) damage. However, the mechanisms involved in the differentiation from tonsil-derived MSCs (T-MSCs) to myocytes via myoblasts remain unclear. To clarify these mechanisms, we analyzed gene expression profiles of T-MSCs during differentiation into myocytes compared with human skeletal muscle cells (hSKMCs). Total RNA was extracted from T-MSCs, T-MSC-derived myoblasts and myocytes, and hSKMCs and was subjected to analysis using a microarray. Microarray analysis of the three phases of myogenic differentiation identified candidate genes associated with myogenic differentiation. The expression pattern of undifferentiated T-MSCs was distinguishable from the myogenic differentiated T-MSCs and hSKMCs. In particular, we selected FNBP1L, which among the upregulated genes is essential for antibacterial autophagy, since autophagy is related to SKM metabolism and myogenesis. T-MSCs differentiated toward myoblasts and skeletal myocytes sequentially, as evidenced by increased expression of autophagy-related markers (including Beclin-1, LC3B and Atg5) and decreased expression of Bcl-2. Furthermore, we reconfirmed that autophagy has an effect on the mechanism of skeletal myogenic differentiation derived from T-MSCs by treatment with 5-azacytidine and bafilomycin A1. These data suggest that the transcriptome of the T-MSC-derived myocytes is similar to that of hSKMCs, and that autophagy has an important role in the mechanism of myogenic differentiation of T-MSCs.</P>

Transplantation of Differentiated Tonsil-Derived Mesenchymal Stem Cells Ameliorates Murine Duchenne Muscular Dystrophy via Autophagy Activation

Park Saeyoung,Jeong Soyeon,Nam Yu Hwa,Yum Yoonji,Jung Sung-Chul 한국조직공학과 재생의학회 2022 조직공학과 재생의학 Vol.19 No.6

BACKGROUND: Skeletal muscles play many important roles in the human body and any malfunction or disorder of the skeletal muscles can lead to a reduced quality of life. Some skeletal dysfunctions are acquired, such as sarcopenia but others are congenital. Duchenne muscular dystrophy (DMD) is one of the most common forms of hereditary muscular dystrophy and is caused by a deficiency of the protein, Dystrophin. Currently, there is no clear treatment for DMD, there are only methods that can alleviate the symptoms of the disease. Mesenchymal stem cells, including tonsil-derived mesenchymal stem cells (TMSCs) have been shown to differentiate into skeletal muscle cells (TMSC-myocyte) and can be one of the resources for the treatment of DMD. Skeletal muscle cell characteristics of TMSC-myocytes have been confirmed through changes in morphology and expression of skeletal muscle markers such as Myogenin, Myf6, and MYH families after differentiation. MEOTHDS: Based on these characteristics, TMSC-myocytes have been transplanted into mdx mice, a mouse model of DMD, to investigate whether they can help improve the symptoms of DMD. The red fluorescent protein gene was transduced into TMSC (TMSC-R) for tracking transplanted cells. RESULTS: Prior to transplantation (TP), it was confirmed whether TMSC-R-myocytes had the same differentiation potential as TMSC-myocytes. Increased expression of dystrophin and autophagy markers in the TP group compared with the sham group was confirmed in the gastrocnemius muscle 12 weeks after TP. CONCLUSION: These results demonstrate muscle regeneration and functional recovery of mdx via autophagy activation following TMSC-myocyte TP.

Differentiation of Human Tonsil-Derived Mesenchymal Stem Cells into Schwann-Like Cells Improves Neuromuscular Function in a Mouse Model of Charcot-Marie-Tooth Disease Type 1A

Park, Saeyoung,Jung, Namhee,Myung, Seoha,Choi, Yoonyoung,Chung, Ki Wha,Choi, Byung-Ok,Jung, Sung-Chul MDPI 2018 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.19 No.8

<P>Charcot-Marie-Tooth disease type 1A (CMT1A) is the most common inherited motor and sensory neuropathy, and is caused by duplication of <I>PMP22</I>, alterations of which are a characteristic feature of demyelination. The clinical phenotype of CMT1A is determined by the degree of axonal loss, and patients suffer from progressive muscle weakness and impaired sensation. Therefore, we investigated the potential of Schwann-like cells differentiated from human tonsil-derived stem cells (T-MSCs) for use in neuromuscular regeneration in trembler-J (Tr-J) mice, a model of CMT1A. After differentiation, we confirmed the increased expression of Schwann cell (SC) markers, including glial fibrillary acidic protein (GFAP), nerve growth factor receptor (NGFR), S100 calcium-binding protein B (S100B), glial cell-derived neurotrophic factor (GDNF), and brain-derived neurotrophic factor (BDNF), which suggests the differentiation of T-MSCs into SCs (T-MSC-SCs). To test their functional efficiency, the T-MSC-SCs were transplanted into the caudal thigh muscle of Tr-J mice. Recipients’ improved locomotive activity on a rotarod test, and their sciatic function index, which suggests that transplanted T-MSC-SCs ameliorated demyelination and atrophy of nerve and muscle in Tr-J mice. Histological and molecular analyses showed the possibility of in situ remyelination by T-MSC-SCs transplantation. These findings demonstrate that the transplantation of heterologous T-MSC-SCs induced neuromuscular regeneration in mice and suggest they could be useful for the therapeutic treatment of patients with CMT1A disease.</P>

Comparison of human first and third trimester placental mesenchymal stem cell.

Park, Saeyoung,Koh, Seong-Eun,Hur, Chang Young,Lee, Won-Don,Lim, Jinho,Lee, Young-Jay Published for the International Federation for Cel 2013 Cell biology international Vol.37 No.3

<P>Placenta mesenchymal stem cells (PMSCs) have the characteristic features of stem cells including renewability in vitro, surface expression, differentiation potency and ability to adhere to the culture surface. PMSCs expressed genes are normally found in the embryonic tissues before the onset of gastrulation, indicating multipotency. However, the stemness can depend on the stages of the placenta from which the cells were isolated. PMSCs were isolated from two different stages of placenta for comparison, that is the first and third trimesters. Both sets had very similar patterns of surface expression as CD44, CD73, CD90 and CD105, and of self renewability in vitro. Expressions of pluripotency-coupled genes were also confirmed in both sets of cells; however, there was a significant difference in the expression levels: fPMSC (mesenchymal stem cells isolated from the first trimester human placenta) being 2-11-fold higher than tPMSC (mesenchymal stem cells isolated from the third trimester human placenta). Possibly due to the difference in the expression levels of the pluripotency-related genes, induction of genes specific to the ectodermal tissues were more prominent in fPMSC than tPMSC after induced differentiation.</P>

Myogenic differentiation potential of human tonsil-derived mesenchymal stem cells and their potential for use to promote skeletal muscle regeneration

PARK, SAEYOUNG,CHOI, YOONYOUNG,JUNG, NAMHEE,YU, YEONSIL,RYU, KYUNG-HA,KIM, HAN SU,JO, INHO,CHOI, BYUNG-OK,JUNG, SUNG-CHUL UNKNOWN 2016 INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE Vol.37 No.5

<P>Stem cells are regarded as an important source of cells which may be used to promote the regeneration of skeletal muscle (SKM) which has been damaged due to defects in the organization of muscle tissue caused by congenital diseases, trauma or tumor removal. In particular, mesenchymal stem cells (MSCs), which require less invasive harvesting techniques, represent a valuable source of cells for stem cell therapy. In the present study, we demonstrated that human tonsil-derived MSCs (T-MSCs) may differentiate into myogenic cells <I>in vitro</I> and that the transplantation of myoblasts and myocytes generated from human T-MSCs mediates the recovery of muscle function <I>in vivo</I>. In order to induce myogenic differentiation, the T-MSC-derived spheres were cultured in Dulbecco's modified Eagle's medium/nutrient mixture F-12 (DMEM/F-12) supplemented with 1 ng/ml transforming growth factor-β, non-essential amino acids and insulin-transferrin-selenium for 4 days followed by culture in myogenic induction medium [low-glucose DMEM containing 2% fetal bovine serum (FBS) and 10 ng/ml insulin-like growth factor 1 (IGF1)] for 14 days. The T-MSCs sequentially differentiated into myoblasts and skeletal myocytes, as evidenced by the increased expression of skeletal myogenesis-related markers [including α-actinin, troponin I type 1 (TNNI1) and myogenin] and the formation of myotubes <I>in vitro</I>. The <I>in situ</I> transplantation of T-MSCs into mice with a partial myectomy of the right gastrocnemius muscle enhanced muscle function, as demonstrated by gait assessment (footprint analysis), and restored the shape of SKM without forming teratomas. Thus, T-MSCs may differentiate into myogenic cells and effectively regenerate SKM following injury. These results demonstrate the therapeutic potential of T-MSCs to promote SKM regeneration following injury.</P>

자율주행자동차 사고의 책임귀속과 교통법제의 정비

박세영(Saeyoung Park) 전남대학교 법학연구소 2022 법학논총 Vol.42 No.4

과학기술의 집합체인 자율주행자동차는 인간을 운전으로부터 자유롭게 할 것이다. 4단계 자율주행시스템부터는 긴급상황에서도 차량 스스로 제어하며 주행하도록 설계되어 있다. 이 단계부터 주행의 동적제어임무(DDT)를 담당하는 기존 운전자 개념은 사라지게 된다. 교통사고의 책임 귀속에 대한 문제가 여전히 남게 되는데, 아직 우리나라 법령은 운행주체나 책임범위에 대한 대비를 충분히 하고 있지 않다. 이 시점에서 영국의 새로운 자율주행자동차법 권고안은 참고할 만한 부분이 많다. 영국정부는 2025년까지 4단계 자율주행자동차의 상용화를 목표로 많은 투자를 진행하고 있다. 이번 권고안은 자율주행자동차와 관련된 수많은 쟁점들을 다루고 있다. 이 가운데 사고책임과 관련하여 눈여겨 볼 만한 부분은 4단계 자율주행자동차의 주체를 두 가지로 구분하고 있다는 것이다. 탑승한 사람이 자동차를 제어할 수 있는지에 따라 각각 유책이용자(有責利用者)와 NUIC 운영자로 구분되고 각각의 자격과 책임은 다르게 인정된다. 운전자는 사고가 발생하면 주행 관련된 사항은 면책되고 책임은 인가된 제조사나 개발자(ASDE)와 NUIC 운영자에게 귀속된다. 제조사(또는 개발자, ASED)나 NUIC 운영자는 면책을 위해 사고에 대해 명백히 다른 원인이 있음을 입증하여야 한다. 그 밖에 운전면허제도와 신호위반, 각종 교통정보의 수집과 저장 등에 관한 교통법제의 근본적인 변화도 모색하여야 한다. Self-driving cars will free humans from driving. Starting with the Level 4 self-driving ADS, is designed to control itself and drive even in emergency circumstances. From now on, the existing concept of a driver in charge of the dynamic driving task(DDT) of driving will be disappeared. The problem of attribution of responsibility for traffic accidents still remains, but the legal situation in Korea has not yet adequately prepared for the subjects or the scope of liability. At this point, the UK's new self-driving car law recommended in 2022 is worth considering. The UK government is investing heavily with the goal of commercializing Level 4 autonomous vehicles by 2025. A number of topics related to autonomous vehicles are dealed with this recommendation. The most remarkable thing in relation to the liability for accidents is that the two subjects of Level 4 autonomous driving vehicles. Depending on whether the person could engage in DDT, it is divided into a user-in-charge and NUIC operator, respectively, and each qualification and responsibility is different. The driver is exempted from driving-related matters in the event of an accident, and the responsibility rests with the manufacturer and operator. For the manufacturer(the developer ASDE) to be indemnified, it would have to be demonstrated by them that there were obviously other causes. It should be sought to fundamental changes in the traffic law system related to the driver's license system, signal violations, and the collection and storage of various traffic information.

Tonsil-Derived Mesenchymal Stem Cells Differentiate into a Schwann Cell Phenotype and Promote Peripheral Nerve Regeneration

Jung, Namhee,Park, Saeyoung,Choi, Yoonyoung,Park, Joo-Won,Hong, Young Bin,Park, Hyun Ho Choi,Yu, Yeonsil,Kwak, Geon,Kim, Han Su,Ryu, Kyung-Ha,Kim, Jae Kwang,Jo, Inho,Choi, Byung-Ok,Jung, Sung-Chul MDPI AG 2016 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.17 No.11

<P>Schwann cells (SCs), which produce neurotropic factors and adhesive molecules, have been reported previously to contribute to structural support and guidance during axonal regeneration; therefore, they are potentially a crucial target in the restoration of injured nervous tissues. Autologous SC transplantation has been performed and has shown promising clinical results for treating nerve injuries and donor site morbidity, and insufficient production of the cells have been considered as a major issue. Here, we performed differentiation of tonsil-derived mesenchymal stem cells (T-MSCs) into SC-like cells (T-MSC-SCs), to evaluate T-MSC-SCs as an alternative to SCs. Using SC markers such as <I>CAD19</I>, <I>GFAP</I>, <I>MBP</I>, <I>NGFR</I>, <I>S100B</I>, and <I>KROX20</I> during quantitative real-time PCR we detected the upregulation of <I>NGFR</I>, <I>S100B</I>, and <I>KROX20</I> and the downregulation of <I>CAD19</I> and <I>MBP</I> at the fully differentiated stage. Furthermore, we found myelination of axons when differentiated SCs were cocultured with mouse dorsal root ganglion neurons. The application of T-MSC-SCs to a mouse model of sciatic nerve injury produced marked improvements in gait and promoted regeneration of damaged nerves. Thus, the transplantation of human T-MSCs might be suitable for assisting in peripheral nerve regeneration.</P>

Digital subtraction angiography vs. real-time fluoroscopy for detection of intravascular injection during transforaminal epidural block

Park, Kibeom,Kim, Saeyoung Yeungnam University College of Medicine 2019 Yeungnam University Journal of Medicine Vol.36 No.2

Background: Transforaminal epidural block (TFEB) is an effective treatment option for radicular pain. To reduce complications from intravascular injection during TFEB, use of imaging modalities such as real-time fluoroscopy (RTF) or digital subtraction angiography (DSA) has been recommended. In this study, we investigated whether DSA improved the detection of intravascular injection during TFEB at the whole spine level compared to RTF. Methods: We prospectively examined 316 patients who underwent TFEB. After confirmation of final needle position using biplanar fluoroscopy, 2 mL of nonionic contrast medium was injected at a rate of 0.5 mL/s under RTF; 30 s later, 2 mL of nonionic contrast medium was injected at a rate of 0.5 mL/s under DSA. Results: Thirty-six intravascular injections were detected for an overall rate of 11.4% using RTF, with 45 detected for a rate of 14.2% using DSA. The detection rate using DSA was statistically different from that using RTF (p=0.004). DSA detected a significantly higher proportion of intravascular injections at the cervical level than at the thoracic (p=0.009) and lumbar (p=0.011) levels. Conclusion: During TFEB at the whole spine level, DSA was better than RTF for the detection of intravascular injection. Special attention is advised for cervical TFEB, because of a significantly higher intravascular injection rate at this level than at other levels.

The evaluation of implementing smart patient controlled analgesic pump with a different infusion rate for different time duration on postoperative pain management

Kim, Saeyoung,Jeon, Younghoon,Lee, Hyeonjun,Lim, Jung A,Park, Sungsik,Kim, Si Oh The Korean Dental Society of Anesthsiology 2016 Journal of Dental Anesthesia and Pain Medicine Vol.16 No.4

Background: Control of postoperative pain is an important aspect of postoperative patient management. Among the methods of postoperative pain control, patient-controlled analgesia (PCA) has been the most commonly used. This study tested the convenience and safety of a PCA method in which the dose adjusted according to time. Methods: This study included 100 patients who had previously undergone orthognathic surgery, discectomy, or total hip arthroplasty, and wished to control their postoperative pain through PCA. In the test group (n = 50), the rate of infusion was changed over time, while in the control group (n = 50), drugs were administered at a fixed rate. Patients' pain scores on the visual analogue scale, number of rescue analgesic infusions, side effects, and patients' satisfaction with analgesia were compared between the two groups. Results: The patients and controls were matched for age, gender, height, weight, and body mass index. No significant difference in the mount of drug administered was found between the test and control groups at 0-24 h after the operation; however, a significant difference was observed at 24-48 h after the operation between the two groups. No difference was found in the postoperative pain score, number of side effects, and patient satisfaction between the two groups. Conclusions: Patient-controlled anesthesia administered at changing rates of infusion has similar numbers of side effects as infusion performed at a fixed rate; however, the former allows for efficient and safe management of postoperative pain even in small doses.

Immunogenicity of COVID-19 Vaccination in Patients With End-Stage Renal Disease Undergoing Maintenance Hemodialysis: The Efficacy of a Mix-and-Match Strategy

Joon-Sung Park,Dohsik Minn,Susie Hong,Saeyoung Jeong,Soohyun Kim,Chang Hwa Lee,Bongyoung Kim 대한의학회 2022 Journal of Korean medical science Vol.37 No.23

Background: The objective of this study was to evaluate the immunogenicity of coronavirus disease 2019 (COVID-19) vaccination in patients with end-stage renal disease (ESRD) on hemodialysis. Methods: ESRD patients at the hemodialysis center of a tertiary-care university-affiliated hospital and healthy employees at the clinical laboratory center were prospectively recruited between March and June 2021. For severe acute respiratory syndrome coronavirus 2 (SARSCoV-2) antibody analysis, blood samples were collected serially on days 0, 14, 28, and 56 after the first vaccine dose, and on days 7 and 50 after the second dose. Antibodies against the SARS-CoV-2 spike protein were quantified, and SARS-CoV-2 neutralizing antibodies were measured in the serum and plasma. Results: Thirty-one ESRD patients and 55 healthy employees were regularly monitored. Twenty-five (80.6%) ESRD patients on hemodialysis received a mix-and-match strategy with ChAdOx1-BNT162b2 (AZ–Pf group) and six (19.4%) received two doses of ChAdOx1 (AZ–AZ group). ESRD patients on hemodialysis showed lower binding antibody titers and neutralizing antibody activities compared to healthy participants following the first vaccination with ChAdOx1. After the second dose, AZ-Pf group had higher immunogenicity than healthy people on days 7 and 50. The binding antibody titer and neutralizing antibody activities on days 7 and 50 were significantly higher in the AZ–Pf group than in the AZ–AZ group. Conclusion: ESRD patients on hemodialysis receiving the mix-and-match strategy (ChAdOx1–BNT162b2) have COVID-19 vaccine immunogenicity comparable to healthy individuals receiving two doses of ChAdOx1.