Kye-Yong Song

Effect of Ascorbic Acid on Bone Marrow-Derived Mesenchymal Stem Cell Proliferation and Differentiation

Mesenchymal stem cells (MSCs) derived from bone marrow are an important tool in tissue engineering and cell-based therapies because of their multipotent capacity. Majority of studies on MSCs have investigated the roles of growth factors, cytokines, and hormones. Antioxidants such as ascorbic acid can be used to expand MSCs while preserving their differentiation ability. Moreover, ascorbic acid can also stimulate MSC proliferation without reciprocal loss of phenotype and differentiation potency. In this study, we evaluated the effects of ascorbic acid on the proliferation, differentiation, extracellular matrix (ECM) secretion of MSCs. The MSCs were cultured in media containing various concentrations (0-500 microM) of L-ascorbate-2-phosphate (Asc-2-P) for 2 weeks, following which they were differentiated into adipocytes and osteoblasts. Ascorbic acid stimulated ECM secretion (collagen and glycosaminoglycan) and cell proliferation. Moreover, the phenotypes of the experimental groups as well as the differentiation potential of MSCs remained unchanged. The apparent absence of decreased cell density or morphologic change is consistent with the toxicity observed with 5-250 microM concentrations of Asc-2-P. The results demonstrate that MSC proliferation or differentiation depends on ascorbic acid concentration.

Molecular cloning and immunological characterization of phosphoglycerate kinase from Clonorchis sinensis

The parasite Clonorchis sinensis was determined to utilize a large amount of external glucose to carry its energy metabolism. Phosphoglycerate kinase (PGK), a glycolytic enzyme, found in many parasites, has been identified as one of the candidate molecules distinguished from human counterparts for vaccine and drug developments. A cDNA clone purified by screening a C. sinensis cDNA library using a heterologous cDNA probe encoded a putative peptide of 415 amino acids with over 60% identities with PGKs from a number of animals. The putative peptides revealed domains corresponding to 12 beta-sheets and inner loops forming a substrate-binding cleft of animal PGKs. The gene product was overexpressed in Escherichia coli and showed a PGK-like enzyme activity. A polyclonal antibody raised against the recombinant C. sinensis PGK was specific to native C. sinensis PGK and localized it to the muscular tissue and tegument of the adult flukes. The C. sinensis PGK elicited antibodies in C. sinensis-infected rabbits. Therefore, it is proposed that C. sinensis PGK could be used as an immunoreagent in the serodiagnosis for clonorchiasis.

Increase in Cell Migration and Angiogenesis in a Composite Silk Scaffold for Tissue-Engineered Ligaments

The purpose of this study was to evaluate the biocompatibility of silk and collagen‐hyaluronan (HA) in vitro by assessing anterior cruciate ligament (ACL) cell and T‐lymphocyte cultures on scaffolds. The use of composite scaffolds as artificial ligaments in ACL reconstruction and their effects on angiogenesis were evaluated in vivo. The silk scaffold was knitted by hand and dry coated with collagen‐HA, whereas the composite silk scaffold was made by covering a silk scaffold with a lyophilized collagen‐HA substrate. The initial attachment and proliferation of human ACL cells on the composite silk scaffold was superior to the attachment and proliferation observed on the silk scaffold. The immune response was higher in both scaffolds after 72 h (p < 0.05) compared with the control culture condition without scaffolding, as assessed by T‐lymphocyte cultures in vitro. There was no significant difference in the immune response in vitro between the silk and composite silk scaffolds. Silk and composite silk scaffolds were implanted as artificial ligaments in ACLs removed from the knees of dogs, and they were harvested 6 weeks after implantation. On gross examination, the onset of an inflammatory tissue reaction, such as synovitis, was seen in both the silk scaffold and the composite silk scaffold groups. An histological evaluation of the artificial ligament implants revealed the presence of monocytes in the silk composite scaffold and the absence of giant cells in all cases. MT staining in the composite silk scaffold‐grafted group showed granulation tissue consisting of fibroblasts, lymphocytes, monocytes, and collagen fibers. In addition, CD31 staining revealed the formation of new blood vessels. On the other hand, no reparative tissues, such as blood vessels, collagen, and cells, were observed in the silk scaffold‐grafted group. These results suggest that the lyophilized collagen‐HA substrate is biocompatible in vitro and enhances new blood vessel and cell migration in vivo.

A intense-focused ultrasound tightening for the treatment of infraorbital laxity

Abstract Background: Infraorbital laxity is a common problem that increases with age. Blepharoplasty with lipectomy is a very commonly performed surgical procedure to treat this problem; however, it is invasive and is associated with the potential for re-emergence. Therefore, young patients may prefer a non-surgical procedure rather than to a surgical procedure. Intense-focused ultrasound (IFUS) has emerged as an effective, non-surgical, tissue-tightening procedure. Objective: This study assessed the safety and efficacy of IFUS (UltheraTM system, Ulthera Inc, Mesa, AZ, USA) for facial tightening in Asian patients with infraorbital laxity. Methods: We studied 15 patients who were treated with an IFUS device applied to both lower eyelids. The primary outcome measure was an objective improvement in a paired comparison of pre-treatment and post treatment (6 months) photographs. A secondary outcome measure was patient satisfaction as measured by a questionnaire. Results: The mean patient age was 50 years (range, 27–69). All patients received one to two treatments with intense-focused ultrasound. All patients in the study experienced both subjective and objective improvement. Conclusion: IFUS can be used as a non-invasive, skin-tightening procedure for infraorbital laxity. No serious, permanent, or delayed side effects were noted up to 6 months post treatment. Thus, this procedure can be effective and safe in the treatment of decreased laxity of the lower eyelids.

Neural stimulation on human bone marrow-derived mesenchymal stem cells by extremely low frequency electromagnetic fields.

Adult stem cells are considered multipotent. Especially, human bone marrow‐derived mesenchymal stem cells (hBM‐MSCs) have the potential to differentiate into nerve type cells. Electromagnetic fields (EMFs) are widely distributed in the environment, and recently there have been many reports on the biological effects of EMFs. hBM‐MSCs are weak and sensitive pluripotent stem cells, therefore extremely low frequency‐electromagnetic fields (ELF‐EMFs) could be affect the changes of biological functions within the cells. In our experiments, ELF‐EMFs inhibited the growth of hBM‐MSCs in 12 days exposure. Their gene level was changed and expression of the neural stem cell marker like nestin was decreased but the neural cell markers like MAP2, NEUROD1, NF‐L, and Tau were induced. In immunofluorescence study, we confirmed the expression of each protein of neural cells. And also both oligodendrocyte and astrocyte related proteins like O4 and GFAP were expressed by ELF‐EMFs. We suggest that EMFs can induce neural differentiation in BM‐MSCs without any chemicals or differentiation factors.

Application of mesenchymal stem cells derived from bone marrow and umbilical cord in human hair multiplication

BACKGROUND The methods currently used for treating alopecia have some limitations. The drug treatment is so temporary that medication discontinuance may progress depilation immediately. The number of hair transplantation restricts because total transplantable hair number is no increase. To overcome these problems, researchers have attempted the in vitro culturing of hair follicle cells and implanting these cells in the treatment area. OBJECTIVES In the present study, culture-expanded mesenchymal stem cells (MSCs) that do not possess aggregative activity were used to produce self-aggregated cell-aggregated spheroidal dermal papilla like tissues (DPLTs) with the aid of a special culture condition in vitro, and hair bulb structure inductive capacity pertinent to the aggregative activity was then evaluated. Then hair inducing activity of self-aggregated DPLTs employing MSCs was tested in athymic mice. METHODS We isolated and cultivated MSCs from bone marrow and umbilical cord in vitro. After propagated MSCs underwent preconditioning in dermal papilla forming medium (DPFM), then subcultured MSCs formed self-aggregated DPLTs. We compared real human scalp dermal papilla cells (hDPCs) with DPLTs employing DPCs, DPLTs employing hBM-MSCs and DPLTs employing hUC-MSCs. RESULTS Light microscopy and immunohistochemical staining were used to confirm that reconstructed DPLTs generated by this procedure had the size, shape, and expression of protein similar to actual DP. CONCLUSIONS The DPLTs have the same hair bulb structure inductive ability as natural DPLTs in vitro. Transplanted DPLTs can induce new hair follicle in athymic mice. As a result, UC-MSCs and BM-MSCs may be an applicable and novel cell source for the generation of human hair cell therapy.

Herbal extract prevents bone loss in ovariectomized rats

This research aims to test a new drug candidate based on a traditional medicinal herb, F1, an herbal extract obtained fromAstragalus membranaceus and its main ingredient, 1-monoli-nolein that may have fewer side effects and less uterine hypertrophy.In vitro experiments, human osteoblast-like cell lines, MG-63 and Saos-2, were analyzed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and an alkaline phosphatase (ALP) assays. Mouse osteoclasts were induced through a calcium-deficient diet and inhibition effects were measured.In vivo experiments were done using ovariectomized (OVX) rats for 9 weeks. At necropsy, uterus weights were measured, trabecular bone area (TBA) of tibia and lumbar vertebra were measured bone histomorphology. In results, cell proliferation and ALP activity in Saos-2 by ether F1 or 1-monolinolein did not increased significantly compared to the control. The F1 inhibited osteoclast development (IC25= 3.37×10−5 mg/mL) less than 17²-estradiol. The OVX rats administered F1 (2 mg/kg/day and 10 mg/kg/day) showed an increase in TBA of the tibia significantly (136.3±4.2% and 138.5±10.3% of control). In conclusions, the herbal extract, F1 inhibited tibia and lumbar bone loss and did not cause uterine hypertrophy. However, 1-monolinolein, the main ingredient of the herbal extract, did not inhibit bone loss.

Treatment of striae distensae combined enhanced penetration platelet-rich plasma and ultrasound after plasma fractional radiofrequency

Abstract Background: Striae distensae (striae alba) is a challenging cosmetic problem for which various treatment modalities have been applied. However, the treatment of striae distensae has not been satisfactory. Objectives: This study was done to evaluate the effectiveness and the safety of enhanced penetration of platelet rich plasma with ultrasound after plasma fractional radiofrequency for the treatment of striae distensae. Subjects and method: Eighteen participants with striae distensae were treated with a Legato system (Alma Lasers, Israel) every two weeks for a total of four sessions. Thereafter, in order to enhance platelet-rich plasma penetration, ultrasound is applied. Clinical photographs were taken before first treatment and two months after the final treatment. Objective and subjective improvement scores were evaluated to demonstrate the efficacy. Abdominal skin biopsies were obtained from three individuals and histological changes were analyzed by light microscopy. Results: During the two months after the last treatment, the average width of the widest striae had decreased from 0.75 to 0.27 mm. In the objective assessment, 71.9% of the participants reported ‘‘good” or “very good’’ overall improvement. In the subjective assessment, and 72.2% of the participants reported ‘‘very satisfied” or “extremely satisfied’’ with overall improvement. The only reported side effect was post-inflammatory hyperpigmentation (11.1%). Conclusions: The plasma fractional radiofrequency and transepidermal delivery of platelet-rich plasma using ultrasound is useful in the treatment of striae distensae.

Hemolytic activity and developmental expression of pore-forming peptide, clonorin

Peptides pore-forming in cell membrane have been identified from a wide range of animals. A putative pore-forming peptide deduced from a cDNA clone of Clonorchis sinensis (clonorin) was predicted to consist of four amphipathic alpha-helices. Clonorin contained six invariably conserved cysteine residues, identified to form three disulfide bonds. These predicted structural features are highly homologous with pore-forming peptides, the amoebapores. Recombinant clonorin showed hemolytic activity toward rabbit erythrocytes. The hemolytic activity of C. sinensis extract increased dose-dependently and was inhibited by anti-clonorin immune sera. The clonorin was expressed developmentally in juvenile and adult flukes and localized in the intestinal epithelium of adult flukes. It is proposed that, through lysing host cellular components, clonorin could enhance proteolytic digestion in the intestine of C. sinensis.

Papular Elastorrhexis in Childhood Improved by Intralesional Injections of Triamcinolone

Papular elastorrhexis is a rare disease developing asymptomatic skin‐colored small papules in adolescence with histopathological loss of elastic fibers. There has been no established treatment for this disease. A 4‐year‐old Korean boy had multiple, hard, whitish papules on his chest and back for one year. Histopathologic examination revealed focal loss of elastic fibers in the dermis, and X‐ray examination showed no bony abnormalities. His skin lesions were improved by intralesional injections of triamcinolone but recurred after four months.