• Journal of Veterinary Clinics
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• v.28 no.5
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• pp.486-496
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• 2011

A special mesenchymal tissue layer called perichondrium has a chondrogenic capacity and is a candidate tissue for engineering of cartilage. To overcome limited potential for chondrocyte proliferation and re-absorption, we studied a method of cartilage tissue engineering comprising chondrocyte-hydrogel pluronic complex (CPC) and cultured perichondrial cell sheet (cPCs) which entirely cover CPC. For effective cartilage regeneration, cell-sheet engineering technique of high-density culture was used for fabrication of cPCs. Hydrogel pluronic as a biomimetic cell carrier used for stable and maintains the chondrocytes. The human cPCs was cultured as a single layer and entirely covered CPC. The tissue engineered constructs were implanted into the dorsal subcutaneous tissue pocket on nude mice (n = 6). CPC without cPCs were used as a controls (N = 6). Engineered cartilage specimens were harvested at 12 weeks after implantation and evaluated with gross morphology and histological examination. Biological analysis was also performed for glycosaminoglycan (GAG) and type II collagen. Indeed, we performed additional in vivo studies of cartilage regeneration using canine large fullthickness chondrial defect model. The dogs were allocated to the experimental groups as treated chondrocyte sheets with perichondrial cell sheet group (n = 4), and chondrocyte sheets only group (n = 4). The histological and biochemical studies performed 12 weeks later as same manners as nude mouse but additional immunofluorescence study. Grossly, the size of cartilage specimen of cPCs covered group was larger than that of the control. On histological examination, the specimen of cPCs covered group showed typical characteristics of cartilage tissue. The contents of GAG and type II collagen were higher in cPCs covered group than that of the control. These studies demonstrated the potential of such CPC/cPCs constructs to support chondrogenesis in vivo. In conclusion, the method of cartilage tissue engineering using cPCs supposed to be an effective method with higher cartilage tissue gain. We suggest a new method of cartilage tissue engineering using cultured perichondrial cell sheet as a promising strategy for cartilage tissue reconstruction.

• Journal of Korean Neurosurgical Society
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• v.48 no.5
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• pp.391-398
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• 2010

Objective : This study was designed to validate the cell trafficking efficiency of the in vivo bioluminescence image (BLI) study in the setting of transplantation of the luciferase expressing bone marrow-derived mesenchymal stem cells (BMSC), which were delivered at each different time after transient middle cerebral artery occlusion (MCAO) in a mouse model. Methods : Transplanting donor BMSC were prepared by primary cell culture from transgenic mouse expressing luciferase (LUC). Transient focal infarcts were induced in 4-6-week-old male nude mice. The experiment mice were divided into five groups by the time of MSC transplantation : 1) sham-operation group, 2) 2-h group, 3) 1-day group, 4) 3-day group, and 5) 1-week group. BLI for detection of spatial distribution of transplanted MSC was performed by detecting emitted photons. Migration of the transplanted cells to the infarcted area was confirmed by histological examinations. Differences between groups were evaluated by paired t-test. Results : A focal spot of bioluminescence was observed at the injection site on the next day after transplantation by Signal intensity of bioluminescence. After 4 weeks, the mean signal intensities of 2-h, 1-day, 3-day, and 1-week group were $2.6{\times}10^7{\pm}7.4{\times}10^6$. $6.1{\times}10^6{\pm}1.2{\times}10^6$, $1.7{\times}10^6{\pm}4.4{\times}10^5$, and $8.9{\times}10^6{\pm}9.5{\times}10^5$, respectively. The 2-h group showed significantly higher signal intensity (p<0.01). The engrafted BMSC showed around the infarct border zones on immunohistochemical examination. The counts of LUC-positive cells revealed the highest number in the 2-h group, in agreement with the results of BLI experiments (p<0.01). Conclusion : In this study, the results suggested that the transplanted BMSC migrated to the infarct border zone in BLI study and the higher signal intensity of LUC-positive cells seen in 2 hrs after MSC transplantation in MCAO mouse model. In addition, noninvasive imaging in real time is an ideal method for tracking stem cell transplantation. This method can be widely applied to various research fields of cell transplantation therapy.

• Korean Journal of Food Science and Technology
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• v.50 no.4
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• pp.451-456
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• 2018

The current study was conducted to confirm the anti-cancer effect of Sarijang, which is a mixture of extracts from purple bamboo salt, Rhynchosia nulubilis, garlic, and Ulmi cortex. Nude mice were injected with a human-derived colorectal cancer cell (HCT116 cell line) and subsequently administered Sarijang for 4 weeks, following which the body weight, organ weight, and tumor size were measured. To evaluate the anti-cancer mechanism of Sarijang, the levels of p16 and extracellular signal-regulated kinase (ERK), cell cycle regulators in colorectal cancer, were measured. To evaluate the toxicity of Sarijang on liver and kidney, aspartate transaminase, alanine transaminase, blood urea nitrogen, and creatinine were analyzed. Sarijang not only reduced the tumor size by enhancing p16 and suppressing ERK, but also showed no side-effect in the liver and kidneys. Taken together, Sarijang has the potential to inhibit tumor growth without side effects, and may be used as a useful functional food.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.41
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• pp.47.1-47.10
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• 2019

Background: Hyaluronic acid (HA) has been applied as a primary biomaterial for temporary soft tissue augmentation and as a carrier for cells and the delivery of growth factors to promote tissue regeneration. Although HA derivatives are the most versatile soft tissue fillers on the market, they are resorbed early, within 3 to 12 months. To overcome their short duration, they can be combined with cells or growth factors. The purpose of this study was to investigate the stimulating effects of human fibroblasts and basic fibroblast growth factors (bFGF) on collagen synthesis during soft tissue augmentation by HA hydrogels and to compare these with the effects of a commercial HA derivative (Restylane^®). Methods: The hydrogel group included four conditions. The first condition consisted of hydrogel (H) alone as a negative control, and the other three conditions were bFGF-containing hydrogel (HB), human fibroblast-containing hydrogel (HF), and human fibroblast/bFGF-containing hydrogel (HBF). In the Restylane^® group (HGF), the hydrogel was replaced with Restylane^® (R, RB, RF, RBF). The gels were implanted subdermally into the back of each nude mouse at four separate sites. Twelve nude mice were used for the hydrogel (n = 6) and Restylane^® groups (n = 6). The specimens were harvested 8 weeks after implantation and assessed histomorphometrically, and collagen synthesis was evaluated by RT-PCR. Results: The hydrogel group showed good biocompatibility with the surrounding tissues and stimulated the formation of a fibrous matrix. HBF and HF showed significantly higher soft tissue synthesis compared to H (p < 0.05), and human collagen type I was well expressed in HB, HF, and HBF; HBF showed the strongest expression. The Restylane^® filler was surrounded by a fibrous capsule without any soft tissue infiltration from the neighboring tissue, and collagen synthesis within the Restylane^® filler could not be observed, even though no inflammatory reactions were observed. Conclusion: This study revealed that HA-based hydrogel alone or hydrogel combined with fibroblasts and/or bFGF can be effectively used for soft tissue augmentation.

• The Korean Journal of Zoology
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• v.31 no.1
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• pp.49-55
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• 1988

A few SV4O-transformed human cells such as SV8O are potentially tumorigenic but rejected by athymic hosts. However, one cell line in this group (W118IVA-2) is known to be fully tumorigenic. Two clones were obtained after the injection of W118IVA-2, of which NW1SC1-1 was tumorigenic but NW18C1-2 was not in nude mice. As examined by Southern blot analysis, NW18C1-1 appears to contain more copy number of SV40 sequences than NW18C1-2 does. However, it was unable to demonstrate that this difference elicits the tumorigenicity in NW18C1-1 but not in NW18C1-2. Therefore, the latter clone was tested if it expresses SV40 early genes to produce large T as well as small t antigens using indirect immunofluorescent assay and immunoprecipitation. In addition, mouse NIH3T3 cells were transfected with the cellular DNA of NW1SC1-2 as well as that of NW18C1-1 to examine if the viral genomes in the clones can make the nontransformed cells to acquire malignant growth potential in vivo. The transformed cells expressed large T antigen and became tumorigenic. Thus, the transforming functions of NW1SC1-2 cell appers to be intact. These results clearly suggest that the inability of NW18C1-2 cell to form tumor in nude mice is not because they are inherently nontumorigenic. However, the possibility that the interaction of SV40 with its host differs in these clones can not he ruled out.

• Journal of Biomedical Engineering Research
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• v.23 no.2
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• pp.147-153
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• 2002

The purpose of this study was to evacuate the effect of different types of Poly(lactic-co-glycolic acid) (PLGA) scaffolds on the formation of human auricular and septal cartilages. All of the scaffolds were formed in a tubular shape for potential application for artificial trachea or esophagus with either 110,000 g/mol PLGA. 220,000 g/mol PLGA. or a combination of both. In order to maintain the tubular shape in vivo, two methods were used. One method was inserting polyethylene tube at the center of scaffolds made of 110,000 g/mol PLGA. The other method involved combination of the two different molecular weight PLGA's. The inner surface of tubular shaped scaffold made with 110,000 g/mol PLGA was coated with 220,000 9/mol PLGA to give more mechanical rigidity. Elastic cartilage was taken from the ear of a patient aged under 20 nears old and hyaline cartilage was taken from the nasal septum. The chondrocytes were then isolated. After second passage, the chondrocytes were seeded on the PLGA scaffolds followed by in vitro culture for one week. The cells-PLGA scaffold complex were implanted subcutaneously on the back of nude mice for 8 weeks. The tissue engineered cartilages were separated from nude mice and examined histologically after staining with the Hematoxylin Eosin. The morphology of the scaffolds were examined by scanning electron microscopy. The pores were well formed and uniformly distributed in the various PLGA scaffolds. After 8 weeks in vivo culture, cartilage was well formed with 110,000 g/mol PLGA. however lumen had collapsed. In contrast. a minimal amount of neocartilage was formed with 220,000 g/mol PLGA, while the architecture of scaffold and lumen were well preserved. Elastic cartilage formed more neocartilage than hyaline. Hyaline and elastic neocartilage were well formed on 110,000 g/mol PLGA with the polyethylene tube, exhibiting mature chondrocytes and preservation of the tubular shape. It was found that 110,000 g/mol PLGA was more appropriate for cartilage formation but higher molecular weight polymer was necessary to maintain the three dimensional shape of the scaffold.

• KSBB Journal
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• v.23 no.4
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• pp.291-296
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• 2008

Amniotic membrane (AM) has been used in various medical application such as biomaterials and it has a biocompatibility and wound healing effects. In this studies, we made AM sponge that was homogenized with AM and then lyophilized. And osteoinduction efficacy of AM sponge was evaluated with collagen sponge by mesenchymal stem cell culture and implantation in nude mouse. As a result of this study, adhesion and proliferation of MSC cells on AM sponge and collagen sponge were not different, but AM sponge was more superior to collagen sponge for induction of collagen secretion and calcium adhesion in matrix in vivo. Besides, AM sponges were more positive stained than collagen sponge about osteocalcin and osteonectin. As a results of this study, there is possibility of doing that AM could increase osteoinduction.

• Journal of Veterinary Clinics
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• v.30 no.6
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• pp.403-408
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• 2013

The objective of the present study is to detect the effect of ${\beta}$-glucan originated from Aureobasidium on the proliferation and collagen production in human dermal fibroblast cells with wound repopulation in vitro. The proliferative effects were assessed using a MTT assay as well as cell counts at 24 and 48 hr after treatment. Hydroxyproline was measured as an index of procollagen production with reverse-phase high pressure liquid chromatography. Oncostatin M was used as a reference agent. In glucagon treated group, dose-dependent and significant increase of optical density or fibroblast cell numbers was demonstrated, when compared with those of control from 0.1 mg/ml concentration. In addition, the numbers of cells which had migrated into the wound defects were more significantly and dose-dependently increased than those of non-treated control. However, no meaningful effects on the procollagen production were observed.

• BMB Reports
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• v.45 no.8
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• pp.470-475
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• 2012

Protein arginine methyltransferase 1 (PRMT1), a type-I arginine methyltransferase, has been implicated in diverse cellular events. We have focused on the role of PRMT1 in gliomagenesis. In this study, we showed that PRMT1 expression was up-regulated in glioma tissues and cell lines compared with normal brain tissues. The knock-down of PRMT1 resulted in an arrest in the G1-S phase of the cell cycle, proliferation inhibition and apoptosis induction in four glioma cell lines (T98G, U87MG, U251, and A172). Moreover, an in vivo study confirmed that the tumor growth in nude mouse xenografts was significantly decreased in the RNAi-PRMT1 group. Additionally, we found that the level of the asymmetric dimethylated modification of H4R3, a substrate of PRMT1, was higher in glioma cells than in normal brain tissues and decreased after PRMT1 knock-down. Our data suggest a potential role for PRMT1 as a novel biomarker of and therapeutic target in gliomas.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.33 no.1
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• pp.1-9
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• 2011

Purpose: Tumor associated angiogenesis and/or lymphangiogenesis are known to be linked by VEGFR signaling pathways. These processes are regulated by several growth factors including VEGFR-2, VEGFR-3. E7080 is an orally active inhibitor of multiple tyrosine kinases including VEGFR-2, 3. Therefore, it was proposed that E7080 may inhibit angiogenesis and lymphangiogenesis. The aim of this study was to determine the effect of E7080 in a nude mouse model of OSCC. Methods: KB cells were xenografted into the submucosal tissue of the mouth floor of athymic mice. Seven days after the xenograft, the mice were randomized into 2 groups. E7080 were administered orally to the experimental group once per day. The mice were sacrificed 3 weeks after the treatment. The tumors were examined histopathologically. Immunohistochemical assays with anti- VEGF-C, VEGFR-2, VEGFR-3, phosphorylated VEGFR-2/3 (pVEGFR-2/3), and D2-40 antibodies were then performed. Results: The transplantation of human OSCC tumor cells into the mouth floor resulted in the formation of orthotopic tumors. The experimental (E7080 treatment) group showed a slowly increased tumor volume. Moreover, immunohistochemical staining demonstrated higher levels of VEGF-C, VEGFR-2, VEGFR-3, pVEGFR-2/3 and D2-40 expression in the control group than in the experimental group. Conclusion: These results suggest that E7080 may provide therapeutic benefits in OSCC.