• Journal of Radiation Protection and Research
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• v.32 no.4
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• pp.140-156
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• 2007

This research was performed to investigate the morphological changes of folliculus ovary according to the radiation dose. The whole body radiation of 200 cGy, 400 cGy, and 600 cGy was given to the each groups of 5 months-aged female mouse. Various staining methods used in this research are: Hematosylin-Eosin method, and immunohistochemistrical methods using BrdU, TUNEL, p53, p21, PCNA and inhibin. The minute structural changes of folliculus ovary were observed through an electron microscope with high magnification. The morphological changes of growing folliculus ovary became distinct as the dose of X-rays increased. Especially, the nuclei of granular cells showed manifest condensation and the changes of the transparent zone were distinct. As a result of histochemical reaction according to Masson's trichrome method and reticular fiber method, the changed granular cells, the deformed basilar membrane of folliculus ovary and the abnormal arrangement of the reticular fiber were observed. In the reaction of BrdU, the granular cells of normal folliculus ovary with positive reaction rapidly decreased according to the increase of the dose of X-rays. In TUNEL study, granular cells showing positive reaction in retarded folliculus ovary were expanded to growing folliculus ovary and primordial folliculus ovary according to the increase of the dose of X-rays. In case of 600 cGy of X-rays, oocyte underwent apoptosis. In p53 immunohistochemistry, p53 manifested to be stronger as the dose of X-rays increased. p53 reactivity was manifested distinctively in all cells comprising folliculus ovary following irradiation of 600 cGy. p21 was manifested in granular cells of folliculus ovary and showed very positive reaction around follicular antrum according to the increase of the dose of X-rays. In PCNA, positive reaction was manifested in growing folliculus ovary, mature folliculus ovary and primordial folliculus ovary, but the extent of the reaction decreased as the dose of the X-rays decreased. The finding that the reaction of granular cells around folliculus ovary was stronger than that near follicular membrane indicates that what was damaged first by X-ray was the cells near folliculus ovary and follicular antrum. The reactivity of $inhibin-{\alpha}$ showed difference according to the growing stage of folliculus ovary: $inhibin-{\alpha}$ showed the most strong reaction in mature folliculus ovary with follicular antrum. There was strong reaction in granular cells around follicular membrane but $inhibin-{\alpha}$ did not occur at all in theca cells comprising follicular membrane. $Inhibin-{\alpha}$ in ovary tissue exposed to 400 cGy of X-rays was manifested more strongly than in ovary tissue exposed to 600 cGy of X-rays, which was related to the phenomenon that granular cells of mature folliculus ovary underwent necrosis or apoptosis increasingly due to X-rays. In an electron microscope with high magnification, nuclei and protoplasm of granular cells in growing folliculus ovary abruptly underwent minute structural changes according to the increase of dose of X-rays. Cell residue, by-product of cell decease, neutrophil and macrophage around follicular antrum were observed. The minute structural changes in granular cells showed typical characteristics of apoptosis: the increase of electronic density due to nuclear condensation, fragmentation of nuclei and atrophy of protoplasm. Necrosis of cells was identified but it was not so remarkable. Macrophage with apoptotic bodies was scattered. Proportional to the radiation dose, we found that the generation of heterogeneous substance of normal ovary texture's follicular fluid, the emergence of dyeing characteristic in the basilar membrane of folicle, the generation of apoptosis, and the transformation of macrophages, etc. From this results, we can infer the possible radiation hazard on the ovary of cervix cancer patient with radiation therapy.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.30 no.2
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• pp.263-278
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• 2004

Ursolic acid (UA) and Oleanolic acid (ONA), known as urson, micromerol and malol, are pentacyclic triterpenoid compounds which naturally occur in a large number of vegetarian foods, medicinal herbs, and plants. They may occur in their free acid form or as aglycones for triterpenoid saponins, which are comprised of a triterpenoid aglycone, linked to one or more sugar moieties. Therefore UA and ONA are similar in pharmacological activity. Lately scientific research, which led to the identification of UA and ONA, revealed that several pharmacological effects, such as antitumor, hepato-protective, anti-inflammatory, anticarcinogenic, antimicrobial, and anti-hyperlipidemic could be attributed to UA and ONA. Here, we introduced the effect of UA and ONA on acutely barrier disrupted and normal hairless mouse skin. To evaluate the effects of UA and ONA on epidermal permeability barrier recovery, both flanks of 8-12 week-old hairless mice were topically treated with either 0.01-0.1mg/mL UA or 0.1-1mg/mL ONA after tape stripping, and TEWL (transepidermal water loss) was measured. The recovery rate increased in those UA or ONA treated groups (0.1mg/mL UA and 0.5mg/mL ONA) at 6h more than 20% compared to vehicle treated group (p < 0.05). Here, we introduced the effects of UA and ONA on acute barrier disruption and normal epidermal permeability barrier function. For verifying the effects of UA and ONA on normal epidermal barrier, hydration and TEWL were measured for 1 and 3 weeks after UA and ONA applications (2mg/mL per day). We also investigated the features of epidermis and dermis using electron microscopy (EM) and light microscopy (LM). Both samples increased hydration compared to vehicle group from 1 week without TEWL alteration (p < 0.005). EM examination using RuO4 and OsO4 fixation revealed that secretion and numbers of lamellar bodies and complete formation of lipid bilayers were most prominent (ONA=UA > vehicle). LM finding showed that thickness of stratum corneum (SC) was slightly increased and especially epidermal thickening and flattening was observed (UA > ONA > vehicle). We also observed that UA and ONA stimulate epidermal keratinocyte differentiation via PPAR Protein expression of involucrin, loricrin, and filaggrin increased at least 2 and 3 fold in HaCaT cells treated with either ONA (10${\mu}$M) or UA (10${\mu}$M) for 24 h respectively. This result suggested that the UA and ONA can improve epidermal permeability barrier function and induce the epidermal keratinocyte differentiation via PPAR Using Masson-trichrome and elastic fiber staining, we observed collagen thickening and elastic fiber elongation by UA and ONA treatments. In vitro results of collagen and elastin synthesis and elastase inhibitory activity measurements were also confirmed in vivo findings. These data suggested that the effects of UA and ONA related to not only epidermal permeability barrier functions but also dermal collagen and elastic fiber synthesis. Taken together, UA and ONA can be relevant candidates to improve epidermal and dermal functions and pertinent agents for cosmeseutical applications.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.3
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• pp.868-873
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• 2004

Renal dysfunction could be developed as the secondary disease of liver cirrhosis. Delayed or suppresed lipid peroxidation by the treatment with physiological active substances could be explained as the antioxidative and protective effect in tissue damage. In this study, we investigated an antioxidative effect and renal function improvement of Hovenia dulcis in liver fibrosis(cirrhosis) induced rats. The female Sprague-Dawley rats (180∼210 g) were divided into 3 groups (Normal, AC: CCl₄ mixture treated group, AC-HV: CCl₄ mixture+ Hovenia dulcis treated group) and renal damage was developed by CCl₄ mixture administration in 4 weeks (0.8 ㎖/rat). The tissue of kidney and liver and sera were used for quantitative measurement of enzyme activity, MDA and Hyp. The histological change and gene expression of collagen α1(III) mRNA and a1(IV) mRNA were observed by Masson's trichrome staining and RT-PCR. As a result, the clinical biochemical parameters of liver function (AST and ALT) in sera of AC-HV group showed significantly 46.4% and 104.8% lower (p<0.005), and the level of ALP and BUN as the parameter of protein urine and azotemia showed 17.8 % and 25.8 % lower than in AC group. In AC-HV group, the concentration of MDA in kidney and liver was decreased significantly 15.8% and 21.3% when compared with AC group (p<0.01 -0.005). The content of Hyp in kidney of AC-HV group is merely higher than in AC group, in contrast to liver tissue. The expression of collagen α1(III) mRNA and collagen α1(IV) mRNA was decreased in AC, but both of collagen mRNA in normal and AC-HV group expressed fast similar. More massive lipid droplets, thicker collagen fiber bundles in portal triads and more formation of portal central septum were observed in the liver of AC group than in AC-HV group. In conclusion, CCl₄ mixture intoxication could be developed not only liver fibrosis(cirrhosis) but also renal dysfunction by the massive lipid peroxidation and suppression of interstitial collagen and basement membrane collagen synthesis. And the water extract of Hovenia dulcis may be possessed the antioxidative and protective effect and improvement of kidney function in renal dysfunction induced rats.

• The korean journal of orthodontics
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• v.27 no.5 s.64
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• pp.825-837
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• 1997

The purpose of this study was to evaluate the effect of splinting methods on the rearrangement of periodontal fibers after experimental tooth movement. Orthodontic force was applied by placing closed coil spring between upper third incisor and canine in seven dogs, weighing 20 kg or more. After 3 weeks of force application, 0.0215 inch multistrand wire and polyethylene ribbon were bonded to each side, as a flexible and rigid splinting respectively in 6 experimental animals. The remaining one served as a control. Each two animals were sacrificed at 4, 8 and 12 weeks after splinting respectively and prepared histologically for hematoxylin-eosin and Masson's trichrome staining. The results of this study were obtained as follows: 1. After tooth movement, periodontal space was narrowed and periodontal titers were thick on pressure side while elongated fibers were observed on tension side. 2. After 4 weeks of retention, the rearrangement of periodontal fibers was observed in the flexible splinting group, but not in the rigid splinting group. 3. After 8 weeks of retention, the rearrangement of periodontal titers was observed in both groups, but the difference could not be detected between two groups. 4. During the retention period, the rearrangement of periodontal fibers was faster in tension side than in pressure side. These results show that the rearrangement of periodontal fibers is also obtained by rigid splinting after tooth movement. It is suggested that the rigid splinting by polyethylene ribbon can be used as a way of postorthodontic retention.

• Korean Journal of Clinical Laboratory Science
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• v.52 no.2
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• pp.112-118
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• 2020

This study was undertaken to examine the effects and to investigate the relevant mechanisms of overexpressing stromal cell-derived factor-1 (SDF-1) produced by engineered mesenchymal stem cells, in a neurogenic bladder (NB) rat model. Sprague-Dawley (SD) rats (N=48) were randomly divided into 4 groups comprising 12 rats each: control group, Injury group, Injury+imMSC group, and Injury+SDF-1 eMSC group. Rats in the Injury+imMSC group were treated with imMSCs, whereas the Injury+SDF-1 eMSC group were administered SDF-1 eMSCs. After 4-weeks therapy, the bladder and pelvic nerve (PN) tissues were examined by subjecting to Masson's trichrome staining and immunofluorescence. Administration of SDF-1 eMSC resulted in improved smooth muscle content in the bladder tissue, significantly increased β-III tubulin expression of the PN, and enhanced SDF-1 expression (P<0.05). The bladder wall repair can be attributed to the overexpression of SDF-1 by SDF-1 eMSCs. Significantly increased SDF-1 expression was obtained in the Injury+SDF-1 eMSC group (P<0.05). The crushed PN also showed significant recovery in the Injury+SDF-1 eMSC group (P<0.05). In conclusion, our results indicate that SDF-1 eMSCs express more SDF-1 in vivo, thereby facilitating the repair of injured nerve and recovery of NB in rats.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.20 no.2
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• pp.112-126
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• 1998

In dentistry, bony defects can be formed by cyst, tumor, inflammation, trauma and surgery in maxilla and mandible. If the overlying soft tissue invades and preoccupies the jaw bony defects, regenerated bony tissue same as adjacent bone can not replace whole space of the defects, thus preventing osteogenesis from occurring. Guided bone regeneration(GBR) is based on the prevention of overlying soft tissue from entering the bony defect during the initial healing periods. E-polytetrafluoroethylene(e-PTFE) is one of an effective and widely used barrier membrane for GBR, but it has the disadvantages such as surgical removal and high price. To overcome such disadvantages of e-PTFE, many investigators have proposed various absorbable barrier membranes. Inexpensive oxidized cellulose($Surgicel^{(R)}$) membrane was shown to have potential for use as an absorbable barrier membrane for regenerative procedure and it would not require surgical removal. The purpose of this study is to investigate the absorption periods of oxidized cellulose at the implant site and usefulness as a mechanical barrier, preventing the ingrowth of the overlying soft tissue into the bony defects. Two bony defects were made in each tibia of a dog using drill and one defect covered with oxidized cellulose and the other covered with periosteum directly as control. The experimental animals were sacrificed at 1st-7th, 10th, 14th, 21th, 28th day postoperatively, Inspection of the specimens was done to evaluate gross changes. Specimens were examined histopathologically by hematoxylin-eosin and Masson's trichrome staining under light microscope. The results were as follows : 1. There was no significant differences of inflammatory reaction between the experimental and the control group. 2. The resorption of oxidized cellulose was almost completed within 14th day. 3. Histologically, bone formation in the experimental group was somewhat more than that of the control group at 10th, 14th, 21th and 28th day postoperatively. The bone forming pattern of the experimental group was more regular than that of the control group. 4. There was no evidence of soft tissue invasion into the bony defect in the experimental group. In conclusion, oxidized cellulose membrane might be used as an alternative absorbable barrier membrane to prevent overlying soft tissue invasion into the bony defects.

• Archives of Reconstructive Microsurgery
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• v.9 no.2
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• pp.139-146
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• 2000

The aim of this study was to assess whether the functional regeneration of a lyophilized autografted cartilage could be improved by implanting a vascularized muscle flap into the medullary canal of autografted proximal humerus. A hemijoint reconstruction using a lyophilized osteochondral autograft in proximal humerus was done in 4 rabbits for control, and combined with an vascularized intramedullary muscle flap in another 4 rabbits for the experimental group. Graft healing and the repair process of osteochondral graft were followed by serial radiographs and histologic changes for 9 weeks after experiments. Each two rabbits in control and in experimental group on 5th and 9th week after implantation of hemijoint were sacrified. The results were as follows: 1. All of control and experimental froups on 5th week united solidly on osteotomized site radiologically, but their articular cartilages were destroyed more seriously in the control than that in experimental group with muscle flap on 5th and 9th week after experiment... 2. Histochemically, the cartilage surface are completely destroyed and revealed with severe osteoarthritic changes on all cartilage layers in control, but cartilaginous erosions are mild to moderate and their arthritic changes are also mild with somewhat regeneration of chondrocytes on deep layers more prominetly on 9th week of the experimental group. 3. The amount of collagen and protenized matrix which was determined by Masson-Trichrome stain was markedly decreased that means the weakness of bony strength and low osteogenic potential in lyophilized cartilage. These results suggest that an intramedullary vascularized muscle flap can improve the functional results of lyophilized osteochondral autograft by providing both increased vascularity and populations of mesenchymal cells to initiate new bone formation on osteotomized site as well as the regeneration of deep layers in articular cartilage. In clinical relevances, this lyophilized hemijoint autograft combined with an intramedullary vascularized muscle pedicle graft might be used very effectively for the treatment of malignant long bone tumors to preserve the joint functions, all or partly, and so to replace it with the artificial joint after tumor excision and hemijoint autograft.

• Journal of Veterinary Clinics
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• v.23 no.3
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• pp.291-299
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• 2006

During long bone lengthening, there are many disadvantages including axial deviation, malalignment and re-fracture which are commonly encountered inspite of its proven abilities. To study the effects of intramedullary K-wire application on the lengthening of long bone, ten skeletally mature mongrel dogs were separated into two groups(Group I, II). Right femurs of group I(5 dogs) were fixed with only monolateral external fixator after subperiosteal osteotomy. Right femurs of group II(5 dogs) were fixed with mono lateral external fixator and intramedullary K-wire after subperiosteal osteotomy. Lengthening was started at 7 days after the surgery with the rate of 0.5 mm per day for 5 weeks and the dogs were sacrificed after 15 weeks postoperatively to examine histologic differences and evaluate bone mineral density. Radiographic examination at an interval of two weeks was done to evaluate the type of callus formed and to analyze complications including instability of external skeletal fixation and axial deviation. Bone mineral density at the lengthened area and contralateral nonlengthened area were measured using quantitative computerized tomography. Histological examination of regenerated bone was performed using Masson's trichrome stain method. The radiographs demonstrated poor callus formation, higher incidence of axial deviation and screw loosening in the group I compared to the group II. The bone mineral density at the lengthened area in the group II was higher than that of the group I(P<0.05). Histological examination showed that the new bone trabeculae in the group II were greater than that of the group I. In conclusion, the combination of monolateral external fixator and intramedullary K-wire can prevent pin loosening, axial deviation and reduce healing period in dogs.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.31 no.5
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• pp.440-453
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• 2005

Objectives : To develop a bioactive membrane for guided bone regeneration (GBR), the biocompatibility and bone regenerating capacity of the cellulose membrane obtained from the Ascidians squirt skin were evaluated. Materials and methods : After processing the pure cellulose membrane from the squirt skin, the morphological study, amino acid analysis and the immunoreactivity of the cellulose membrane were tested. Total eighteen male Spraque-Dawley rats (12 weeks, weighing 250 to 300g) were divided into two control (n=8) and another two experimental groups (n=10). In the first experimental group (n=5), the cellulose membrane was applicated to the 8.0 mm sized calvarial bone defect and the same sized defect was left without cellulose membrane in the first control group (n=4). In the another experimental group (n=5), the cellulose membrane was applicated to the same sized calvarial bone defect after femoral bone graft and the same sized defect with bone graft was left without cellulose membrane in the another control group (n=4). Each group was sacrificed after 6 weeks, the histological study with H&E and Masson trichrome stain was done, and immunohistochemical stainings of angiogenin and VEGF were also carried out. Results : The squirt skin cellulose showed the bio-inductive effect on the bone and mesenchymal tissues in the periosteum of rat calvarial bone. This phenomenon was found only in the inner surface of the cellulose membrane after 6 weeks contrast to the outer surface. Bone defect covered with the bioactive cellulose membrane showed significantly greater bone formation compared with control groups. Mesenchymal cells beneath the inner surface of the bioactive cellulose membrane were positive to the angiogenin and VEGF antibodies. Conclusion : We suppose that there still remains extremely little amount of peptide fragment derived from the basement membrane matrix proteins of squirt skin, which is a kind of anchoring protein composed of glycocalyx. This composition could prevent the adverse immunological hypersensitivity and also induce bioactive properties of cellulose membrane. These properties induced the effective angiogenesis with rapid osteogenesis beneath the inner surface of cellulose membrane, and so the possibilities of clinical application in dental field as a GBR material will be able to be suggested.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.29 no.2
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• pp.123-131
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• 2007

Introduction: Distraction osteogenesis is widely used as for bone lengthening in patients with maxillofacial deformity and alveolar bone atrophy. One of the major problems in distraction osteogenesis is long consolidation period for 2-3 months, in which the devices have to be fixed on the bone to prevent relapse. It results in scar formation on the face, disturbance of mastication and speech. This study was performed to evaluate the stimulating effect of pulsed electromagnetic field on the early bone consolidation in distraction osteogenesis. Materials and methods: Total 10 rabbit were used (5 for control group, 5 for experimental group). A vertical osteotomy in the mandibular body was performed and the distraction device was fixed. After 5 days distraction was done 1mm per a day for 7 days. A pulsed electromagnetic field (38 Gauss, 60 Hz) was applied for 8 hours per day and it continued for 5 days immediately after distraction in the experimental group. Both groups were sacrificed after 2 weeks. Histological specimens with H&E and Masson Trichrome staining were made and histomorphometrically analysed with image analyser. Results: The device for distraction osteogenesis was displaced in one animal for each group, therefore, only four animals in both groups were evaluated. In both groups, a new bone formation was observed in the distracted area after 2 weeks. The bone formation was enhanced in the experimental groups ($31.76{\pm}8.68%$) compared with control group ($9.94{\pm}3.23%$), its difference was statistically significant (p<0.001). Conclusion: This study suggests that electrical stimulation with electromagnectic field may be effective in the early bone formation after distraction osteogenesis. Further studies with large number of animals are needed before clinical application.