• Journal of Microbiology and Biotechnology
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• v.15 no.2
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• pp.259-264
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• 2005

Transplantation of cultured chondrocytes can regenerate cartilage tissues in cartilage defects in humans. However, this method requires a long culture period to expand chondrocytes to a large number of cells for transplantation. In addition, chondrocytes may dedifferentiate during long-term culture. These problems can potentially be overcome by the use of undifferentiated or partially developed cartilage precursor cells derived from neonatal cartilage, which, unlike chondrocytes from adult cartilage, have the capacity for rapid in vitro cell expansion and may retain their differentiated phenotype during long-term culture. The purpose of this study was to compare the cell growth rate and phenotypic modulation during in vitro culture between adult chondrocytes and neonatal chondrocytes, and to demonstrate the feasibility of regenerating cartilage tissues in vivo by transplantation of neonatal chondrocytes expanded in vitro and seeded onto polymer scaffolds. When cultured in vitro, chondrocytes isolated from neonatal (immediately postpartum, 2 h of age) rats exhibited much higher growth rate than chondrocytes isolated from adult rats. After 5 days of culture, more neonatal chondrocytes were in the differentiated state than adult chondrocytes. Cultured neonatal chondrocytes were seeded onto biodegradable polymer scaffolds and transplanted into athymic mice's subcutaneous sites. Four weeks after implantation, neonatal chondrocyte-seeded scaffolds formed white cartilaginous tissues. Histological analysis of the implants with hematoxylin and eosin showed mature and well-formed cartilage. Alcian blue/ safranin-O staining and Masson's trichrome staining indicated the presence of highly sulfated glycosarninoglycans and collagen, respectively, both of which are the major extracellular matrices of cartilage. Immunohistochemical analysis showed that the collagen was mainly type II, the major collagen type in cartilage. These results showed that neonatal chondrocytes have potential to be a cell source for cartilage tissue engineering.

• The korean journal of orthodontics
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• v.19 no.2
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• pp.57-73
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• 1989

The purpose of this study was to investigate the tissue response of the rat molar periodontium incident to intermittent orthodontic force. The author intended to observe the healing process of injured periodontium and the response of injured tissue to the resumed force. Oxytetracyclin 50mg/Kg was given to each rat intraperitonially. 5 days later, maxillary 1st molars were moved mesially from the incisors with closed coil spring of 100gram. 7 days later, the appliances were removed and 20mg/Kg of calcein were given intraperitonially to each rat. At the same time, maxillary left 1st molars of 15 rats were moved by the same method, but force was lowered to 20 gram. After 1 day, maxillary left 1st molars of another 15 rats were moved by the same method and 50mg/Kg of oxytetracycline was given intraperitonially. After 4 days, another 15 rats were treated as above. After 7 days, another 15 rats were treated as above. 1,4,7,10 and 14 days after change of force, 3 rats were sacrificed in each group respectively. 2 rats were decalcified, embedded in paraffin, and stained with hematoxylin-eosin stain and with Masson's trichrome stain. Another rat was embedded in polyester resin and undecalcified specimen were made. Microradiograms were taken with the undecalcified sections. Observations were made with light and fluorescence microscope. Following conclusions were made. 1. Connective tissue cells and vessels were infiltrated into the hyalinized tissue from the bony cleft and along the border of the hyalinized tissue with bone and root surface. At the same time, elimination of hyalinized tissue, bone and root resorption occurred. 2. Bone and root were resorbed directly and indirectly. 3. Hyalinized tissue was removed within 5 days after force removal. 4. Hyalinized zone was less extensive and easily removed as the rest period prolonged. 5. Hyalinized tissue developed more rapidly and extensively and lasted over 10 days as the force resumed on the already formed hyalinized tissue.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.35 no.1
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• pp.13-17
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• 2013

Purpose: This study evaluated the capability of bone formation of silk fibroin particles coated with hydroxyapatites (HA/SF), as bone graft material when put into the calvarial defect of rats. Methods: Twenty Sprague Dawley rats were used for this study and round shaped defects were formed in the center of parietal bones (diameter: 8.0 mm). The defect was filled with (1) HA/SF (experimental group), or (2) left as a vacant space (control group). The animals were sacrificed at 4 or 8 weeks, postoperatively. The specimens were decalcified and stained with Masson's trichrome for histomorphometric analysis. Results: The average of new bone formation was $33.18{\pm}3.10%$ in the experimental group and $20.49{\pm}5.79%$ in the control group at 4 weeks postoperatively. That was $42.52{\pm}7.74%$ in the experimental group and $25.50{\pm}7.31%$ in the control group at 8 weeks postoperatively. The difference between the groups was significantly higher at both 4 weeks and 8 weeks postoperatively (P<0.05). Conclusion: The rat calvarial defect was successfully repaired by HA/SF graft. The HA/SF graft showed more new bone formation compared with the unfilled control.

• The korean journal of orthodontics
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• v.14 no.1
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• pp.53-65
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• 1984

The purpose of this study was to investigate the histopathological change and adaptation process of the mandibular joint of the rat by muscle alteration. For this study, one hundred and twenty eight rats of 25 - and 60 - day old of age were used. Unilateral and bilateral detachment, with anterior positioning of the Masseter muscle, was performed under anesthesia. The animal was sacrified 10, 20, 50, 80 days postoperatively. This alteration in muscle function led to change in neuromuscular activity and demonstrated the adaptive nature of the condyle cartilage to functional demand. The results were as follows : 1. In the right muscle detached group, operated at 25 days of age, marked decrease on the chondroblastic zone was found in the condyle head on the right side of animals examined 10 days postoperatively. Comparing with the control group, no difference was found on the chondroblastic zone in the condylar head of animals examined 20, 50 and 80 days postoperatively. 2. In the bilateral muscle detached group, operated at 25 days of age, the chondroblastic zone was slightly decreased in the anterior parts of condylar head of animals examined 10 days postoperatively. 3. In the unilateral and bilateral muscle detached group, operated at 60 days of age, no significant change was found in the mandibular joint regardless of the post operative experimental periods. 4. Under Toluidine blue staining, slightly decreased metachromasia was found in the condyle head on the right side of unilateral experimental animals, operated at 25 days of age and examined 10 days postoperatively. 5. Under Masson's trichrome staining, increased metachromasia was found in the condyle head on the right side of unilateral experimental animals, operated at 25 days of age and examined 10 days postoperatively. In summary, the condyle of the rat could respond to changes in neuromuscular activity depend on the level of maturation of the tissue, because the endochondral bone formation of the condyle of the rat was almost ended within 3 months.

• Biomedical Science Letters
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• v.13 no.1
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• pp.55-60
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• 2007

Computerized image processing and analysis system was used for quantitative assessment of skin tissue components in color histological sections. The purpose of this study was to determine that the changes in the collagen fiber density and elastic fiber density in dermis in the rat skin as aging. And also to determine that the correlation between fibroblast density and collagen fiber density and elastic fiber density in the aged rat skin. Ten weeks old ($130{\sim}150g$) eight and fifty-four weeks old ($300{\sim}350g$) eight female Sprague-Dawley rats were used. The full-thickness skin biopsy specimens were prepared serial sections and stained with hematoxylin and eosin, Masson's trichrome and Verhoeff-van Gieson. The collagen fiber and the elastic fiber were identified using the image analysis processing system and then calculated the collagen fiber density rate and the elastic fiber density rate in the dermis. It also identified fibroblast and calculated fibroblast density in the dermis. By using a Student's t-test, a decrease in the collagen fiber density rate (t=-4.650, P<0.001) and the elastic fiber density rate (t=-6.494, P<0.001) of dermis can be observed in aged rats as compared with the young rats. A Student's t-test showed a significantly less fibroblast density in the aged rats than the young rats (t=-4.497, P<0.001). There were significantly positive correlation between the fibroblast density and the collagen fiber density rate (r=.69, P<0.001) and the elastic fiber density rate (r=.91, P<0.001). These results indicate that the aging may decrease the collagen fiber density and elastic fiber density due to reduced the proliferative and synthetic activity of fibroblast in the dermis.

• Clinical and Experimental Pediatrics
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• v.59 no.6
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• pp.262-270
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• 2016

Purpose: Pulmonary arterial hypertension (PAH) leads to right ventricular failure (RVF) as well as an increase in pulmonary vascular resistance. Our purpose was to study the effect of sildenafil on right ventricular remodeling in a rat model of monocrotaline (MCT)-induced RVF. Methods: The rats were distributed randomly into 3 groups. The control (C) group, the monocrotaline (M) group (MCT 60 mg/kg) and the sildenafil (S) group (MCT 60 mg/kg+ sildenafil 30 mg/kg/day for 28 days). Masson Trichrome staining was used for heart tissues. Western blot analysis and immunohistochemical staining were performed. Results: The mean right ventricular pressure (RVP) was significantly lower in the S group at weeks 1, 2, and 4. The number of intra-acinar arteries and the medial wall thickness of the pulmonary arterioles significantly lessened in the S group at week 4. The collagen content also decreased in heart tissues in the S group at week 4. Protein expression levels of B-cell lymphoma-2 (Bcl-2)-associated X, caspase-3, Bcl-2, interleukin (IL)-6, matrix metalloproteinase (MMP)-2, endothelial nitric oxide synthase (eNOS), endothelin (ET)-1 and ET receptor A (ERA) in lung tissues greatly decreased in the S group at week 4 according to immunohistochemical staining. According to Western blotting, protein expression levels of troponin I, brain natriuretic peptide, caspase-3, Bcl-2, tumor necrosis factor-${\alpha}$, IL-6, MMP-2, eNOS, ET-1, and ERA in heart tissues greatly diminished in the S group at week 4. Conclusion: Sildenafil alleviated right ventricular hypertrophy and mean RVP. These data suggest that sildenafil improves right ventricular function.

• Biomedical Science Letters
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• v.16 no.4
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• pp.365-376
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• 2010

The purpose of this study was to clarify the effect of light emitting diode (LED) irradiation on healing of impaired wound and alteration of mast cells in experimental diabetic rats. Twenty-four male Sprague-Dawley rats were divided into four groups: excision (Ex), excision-LED irradiation (Ex-LED), diabetes + excision (DM) and diabetes + excision + LED irradiation (DM-LED). Diabetes was induced in rats by streptozotocin (STZ) injection (70 mg/kg, single dose) and 6 mm punch excision wounds were created on the back after shaving hair. The LED-irradiated rats were treated to a daily dose of $5\;J/cm^2$ LED (630 nm) light for 11 days after surgery, and were killed at day 1, 3, 7 and 11. The lesion and adjacent skin tissues were excised, fixed with 10% buffered formalin and embedded with paraffin. For evaluation of wound healing, hematoxylin-eosin (HE) and Masson trichrome staining were performed. Mast cells (MCs) were stained with toluidine blue (pH 0.5) and quantified using a computerized image analysis system. The proliferation activity of keratinocyte in skin tissues was analyzed on sections immunostained with proliferative cell nuclear antigen (PCNA). The results showed that wound healing rate, collagen density and neo-epidermis length, number of PCNA-positive cells, fibroblasts and mast cells were significantly higher in the LED-irradiated rats than in the DM and Ex rats throughout the periods of experiment. Exceptionally, the number of MCs was significantly lower at day 11 compared with day 7 after surgery in the all groups. These findings suggest that the LED irradiation may promote the tissue repair process by accelerating keratinocyte and fibroblast proliferation and collagen production in normal rats as well as in diabetic rats, and MCs may play an important role at an early stage of skin wound healing in normal and diabetic rats.

• Current Optics and Photonics
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• v.3 no.6
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• pp.485-495
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• 2019

Photobiomodulation (PBM) using organic light emitting diodes (OLEDs) surface light sources have recently been claimed to be the next generation of PBM light sources. However, the differences between light emitting diodes (LEDs) and OLED mechanisms in vitro and in vivo have not been well studied. In vivo mouse models were used to investigate the effects of OLED irradiation on cellular function and cutaneous wound healing compared to LED irradiation. Mice in the LED- and OLED-irradiated groups were subjected to irradiation with 6 J/㎠ LED and OLED (630 nm), respectively, for 14 days after wounding, and some mice were sacrificed for the experiments on days 3, 7, 10, and 14. To evaluate wound healing, we performed hematoxylin-eosin and Masson's trichrome staining and quantified collagen density by computerized image analysis. The results showed that the size of the wound, collagen density, neo-epidermis thickness, number of new blood vessels, and number of fibroblasts and neutrophils was significantly influenced by LED and OLED irradiation. The tissue levels of interleukin (IL)-β, IL-6 and tumor necrosis factor (TNF)-α were investigated by immunohistochemical staining. LED and OLED irradiation resulted in a significant increase in the tissue IL-β and IL-6 levels at the early stage of wound healing (P < 0.01), and a decrease in the tissue TNF-α level at all stages of wound healing (P < 0.05), compared to the no-treatment group. The expression levels of the genes encoding vascular endothelial growth factor and transforming growth factor-beta 1 were significantly increased in LED and OLED-irradiated wound tissue at the early stage of wound healing (P < 0.01) compared to the no-treatment group. Thus, OLED as well as LED irradiation accelerated wound healing by modulating the synthesis of anti-inflammatory cytokines and the expression levels of genes encoding growth factors, promoting collagen regeneration and reducing scarring. In conclusion, this suggests the possibility of OLED as a new light source to overcome the limitations of existing PBMs.

• Development and Reproduction
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• v.19 no.3
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• pp.145-152
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• 2015

Diphlorethohydroxycarmalol (DPHC) is a known to modulate the expression of extracellular matrix (ECM) components in 3T3-L1. However, the possible role of DPHC in integument stability during obesity induction is not clear yet. We evaluated the effects of DPHC on collagen or elastic fiber quantity in integument during obesity induction with high-fat diet. The dorsal back integument sections were stained with hematoxylin-eosin, Masson trichrome, and Verhoff-Van Gieson. The intensities of collagen fibers and elastin fibers were analyzed with ImageJ. The number of fibroblasts was counted at ${\times}1,000$ fields. The number of fibroblast was increased by obesity induction, but DPHC suppressed it in a concentration-dependent manner both in lean and obese mice. On the other hand, the intensities of collagen fibers were increased by DPHC treatment in obese mice groups but not in lean mice groups. The intensities of collagen fibers of obese mice were lower than that of the lean mice in 0% group. However, the number became similar between lean and obese mice by the treatment of DPHC. The intensity of elastic fibers was increased in the lean mice with the concentration of DPHC. In the obese mice group, there were increasing patterns but only significant at 10% DPHC group. The intensity of elastic fibers of obese mice was higher than lean mice in 0%, 1%, and 10% groups. Histologically epithelial cells and follicle cells which were diffused nuclear staining forms were increased by DPHC treatment. The results suggest that the activity of integument cells during obesity induction can be modulated by DPHC.

• Journal of Physiology & Pathology in Korean Medicine
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• v.26 no.5
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• pp.732-737
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• 2012

The aim of this study is examine the protection effect for artery and antithrombotic effect of Sokmyong-tang(SMT; xuming-tang) on carotid artery thrombosis in a rat model. Thirty minutes before $FeCl_3$ treatment, SD rats were intraperitoneal injected with SMT. We tested the effects of SMT on time to occlusion (TTO) in thrombosis model by induced $FeCl_3$ using the laser Doppler flow meter and accessed thrombus weight (TW) inhibition and measured collagen fibers in the vessel after injury using Masson's trichrome stain. SMT(100 mg/kg, i.p.) showed significantly delayed TTO ($13.17{\pm}1.33$ min, P < 0.001) compared to vehicle control group ($8.63{\pm}0.92$ min) and inhibiting effect on TW with $0.72{\pm}0.02$ mg/mm (P < 0.05). In addition, SMT prevented collagen fibres damage in injured vessel ($22.24{\pm}4.48%$, P < 0.001). These results provide experimental evidence for SMT can be used to prevent vascular injury and thrombosis such as hypertension, arteriosclerosis, and so on.