• Title/Summary/Keyword: Masson's trichrome stain

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Pine bark extract (Pycnogenol®) suppresses cigarette smoke-induced fibrotic response via transforming growth factor-β1/Smad family member 2/3 signaling

  • Ko, Je-Won;Shin, Na-Rae;Park, Sung-Hyeuk;Kim, Joong-Sun;Cho, Young-Kwon;Kim, Jong-Choon;Shin, In-Sik;Shin, Dong-Ho
    • Laboraroty Animal Research
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    • v.33 no.2
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    • pp.76-83
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    • 2017
  • Chronic obstructive pulmonary diseases (COPD) is an important disease featured as intense inflammation, protease imbalance, and air flow limitation and mainly induced by cigarette smoke (CS). In present study, we explored the effects of $Pycnogenol^{(R)}$ (PYC, pine bark extract) on pulmonary fibrosis caused by CS+lipopolysaccharide (LPS) exposure. Mice were treated with LPS intranasally on day 12 and 26, followed by CS exposure for 1 h/day (8 cigarettes per day) for 4 weeks. One hour before CS exposure, 10 and 20 mg/kg of PYC were administered by oral gavage for 4 weeks. PYC effectively reduced the number of inflammatory cells and proinflammatory mediators caused by CS+LPS exposure in bronchoalveolar lavage fluid. PYC inhibited the collagen deposition on lung tissue caused by CS+LPS exposure, as evidenced by Masson's trichrome stain. Furthermore, transforming growth $factor-{\beta}1$ ($TGF-{\beta}1$) expression and Smad family member 2/3 (Smad 2/3) phosphorylation were effectively suppressed by PYC treatment. PYC markedly reduced the collagen deposition caused by CS+LPS exposure, which was closely involved in $TGF-{\beta}1$/Smad 2/3 signaling, which is associated with pulmonary fibrotic change. These findings suggest that treatment with PYC could be a therapeutic strategy for controlling COPD progression.

Tributyltin chloride (TBTCl) toxicity on the survival rate and burrowing activity of the equilateral venus, Gomphina veneriformis (Bivalvia: Veneridae) (대복, Gomphina veneriformis의 생존 및 저질 잠입성에 미치는 TBTCl의 독성)

  • Park, Jung-Jun;Jin, Young-Guk;Lee, Jung-Sick
    • Journal of fish pathology
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    • v.21 no.1
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    • pp.45-56
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    • 2008
  • This study was conducted to find out biological responses of bivalves exposed to organotin compound.The results of the study confirmed that tribultyltin chloride (TBTCl) induce reduction of survival rate andburrowing activity, and histopathological feature in the foot structure of the equilateral venus, Gomphinaveneriformis. The experimental period was 36 weeks. The experimental groups consisted of a control and 3TBTCl exposure groups (0.4, 0.6, 0.8 ym TBTCl L'). The survival rate and burrowing activity were record-ed daily. For histological analysis, foot tissues were fixed in Bouin' s fluid and then stained H-E stain, AB-PAS (PH 2.5) reaction and Masson's trichrome stain after having serially sectioned the tissue by paraffinmethod at thickness of 4-6 ym. The survival rate was not significantly different between the control andexposure groups for 20 weeks, but in 0.8 Um TBTCl L', it was on the decreased ever since the exposure. Theburrowing activity was not significantly different in the exposure group compared to the control up to 12weeks, but in 0.6 and 0.8 ym TBTCl L', it measured the lowest level after 20 weeks. The foot is composedof the epidermal layer, connective tissue, and muscular layer. The epidermal layer is composed of simplecolumnar, cuboidal epithelia and mucous cells. The cilia were well developed on the apical surface ofepithelium, Circular, longitudinal and transverse muscle bundle were well developed in the muscular layer.The majority mucous cells showed blue color (542c) when it subjected to AB-PAS (PH 2.5) reaction. Nohistopathological alterations in the foot were observed up to 12 weeks. After 20 weeks of exposure to 0.8 (anTBTCl L'', the foot samples of exposed G. veneriformis showed disappearance of cilia and striated borderpartially and extension of hemolymph sinus. The mucous cell increased in the marginal of foot. At 28-weekof exposure to 0.4 ym TBTCl L', it observed weekly acid (564c), neutral (264c) and mixed mucous cell. At36-week of exposure to 0.6 ym TBTCl L', it showed fragmentation of the muscle and collagen fiber bundle,and also diappearance of cilia on epithelia and edema of epithelium in 0.8 ym TBTCl L''.

The Effects of Bone Morphogenetic Protein-4 and Resorbable Membrane on the Regeneration of Periodontal Tissues (골형태형성단백질과 흡수성차폐막이 치주조직 재생에 미치는 영향)

  • Lim, Sang-Cheol;Kwon, Young-Hyuk;Lee, Man-Sup;Partk, Joon-Bong
    • Journal of Periodontal and Implant Science
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    • v.30 no.4
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    • pp.757-779
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    • 2000
  • The aim of our study is to achieve complete periodontal tissue regeneration by the application of BMP and resorbable membrane. Three beagle dogs aged over one and half years and weighed 14 to 16 kg were used in this study. Mandibular 1st, 2nd premolars were extracted bilaterally. Horizontal furcation defects were induced around 3rd, 4th premolars bilaterally. BMP-4 were applied in the right side with resorbable membranes and only resorbable membranes were applied in the left side respectively. Each animal was sacrificed at 2, 4, and 8weeks, after regenerative surgery. Specimens were prepared with Hematoxylin-Eosin stain and Goldner's modified Masson Trichrome stain for light microscopic evaluation. The results were as follows: 1. At 2 weeks after regenerative surgery, downgrowth of junctional epithelium was observed both in the membraneapplied site and BMP-4-and-membrane-applied site. 2. At 4 weeks after regenerative surgery, resorbable membranes were completely resolved, therefore would not prevent downgrowth of junctional epithelium. New bone formation, new cementum formation and Sharpey's fiber were observed in BMP-4-andmembrane-applied site. 3. At 8 weeks after regenerative surgery, downgrowth of junctional epithelium was observed in the membrane-applied site. But, new cementum formation was observed in the same site. The extensive regeneration of new bone, new cementum and remarkable formation of Shapey's fiber were showed in BMP-4-and-membrane-applied site. 4. Resorbable membranes were resolved via the cell-mediated processes. 5. Periodontal tissue regeneration were better achieved in the BMP-4-andmembrane-applied site than in the membrane-applied site. Within the above results, BMP-4 may have the strong capability to form the new bone and resorbable membrane may be able to prevent the bony ankylosis. However, resolution rate of resorbable membrane may not be enough to protect rapid epithelial downgrowth for ideal periodontal regeneration. In conclusion, I suggest BMP-4 may have the strong possibility to be utilized in the clinical periodontal treat-

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Guided Tissue Regeneration Using Barrier Membrane and Osseous Grafts in Surgically Created Furcation Defects in Dogs (성견의 외과적 치근이개부 골결손에 차폐막과 골이식재를 이용한 조직유도재생술시 치유양상)

  • Chung, Eun-Hee;Chung, Hyun-Ju
    • Journal of Periodontal and Implant Science
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    • v.26 no.4
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    • pp.967-987
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    • 1996
  • The present study was to evaluate the healing patterns of guided tissue regeneration( GTR) using resorbable $Vicryl^{(R)}$(polyglactin 910) mesh and nonresorbable expanded polytetrafluoroethylene(ePTFE) membrane with or without bone grafting using autogeneous bone and demineralized freeze-dried bone allograft(DFDBA) in the grade II furcation defects. Mucoperiosteal flaps were reflected buccally in the mandibular 2nd, 3rd and 4th premolar areas and furcation defects were created surgically by removing $5{\times}6mm$ alveolar bone in 4 dogs. Root surfaces were thoroughly debrided of periodontal ligament and cementum, and notches were placed on root surface at the most apical bone level. In the right and left mandibular quadrant, each tooth was received $Vicryl^{(R)}$ mesh(ACE Surgical Supply Co., USA) only, $Vicryl^{(R)}$ mesh with DFDBA, $Vicryl^{(R)}$ mesh with autogeneous bone grafts, ePTFE membrane($Core-tex^{(R)}$ membrane, W.L. Gore & Associates Inc., USA) only, ePTFE membrane with DFDBA or ePTFE membrane with autogeneous bone grafts. For the fluorescent microscopic examination, fluorescent agents were injected at 2, 4 and 8 weeks after surgery. Four weeks after surgery, 2 dogs were sacrificed and ePTFE membranes were removed from remaining 2 dogs, which were sacrificed at 12 weeks after surgery. Undecalcified tissues were embedded in methylmethacrylate and $10{\mu}m$ thick sections were cut in a buccolingual direction. These sections were stained with hematoxylin-eosin stain and Masson's trichrome stain, and evaluated by descriptive histology and linear measurements. The results were as follows : 1) $Vicryl^{(R)}$ mesh group showed less connective tissue attachment than ePTFE membrane group. 2) The combination of GTR using $Vicryl^{(R)}$ mesh and osseous grafts resulted in new attachment and new bone formation more than GTR using $Vicryl^{(R)}$ mesh only. 3) GTR using ePTFE membrane, with or without osseous grafts, enhanced periodontal regeneration. 4) Root resorption and dentoalveolar ankylosis were observed in the areas treated with the combination of GTR and DFDBA. It was suggested that the effect of adjunctive bone grafting in GTR procedure depends on the materials and the physical properties of barrier membranes. $Vicryl^{(R)}$ mesh performed a barrier function and the use of adjunctive bone grafting may enhance the periodontal regeneration.

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Suppressive Effect of Kwanjulbang-6 on Collagen-Induced Arthritis in Mice (관절6호방이 Type II collagen으로 유발된 관절염에 미치는 영향)

  • Kim, Hyun-Soo;Hong, Seo-Young;Heo, Dong-Seok;Oh, Min-Suck
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.5
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    • pp.1003-1011
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    • 2009
  • This study was carried out to know the effects of Kwanjulbang-6(GJB-6) on the inhibition of arthritis. GJB-6 was orally administered to mouse with arthritis induced by collagen II . Cytotoxicity, hepatotoxicity, arthritis index, value of immunocyte in draining lymph node and paw joint, rheumatoid factor in serum were measured in vivo. The incidence of arthritis was significantly decreased. Total cell number of draining lymph node was significantly increased compared with control. Total cell number of paw joint was significantly decreased compared with control. The absolute number of $CD19^+$, $CD8^+$, $CD3^+/CD69^+$, $CD3^+/CD49b^+$, $CD4^+/CD44^+$, $CD3^+$, $CD4^+$, $CD4^+/CD25^+$ and $CD3^+/CD8^+$ cells in draining lymph node were significantly increased compared with control. The absolute number of $CD3^+$, $CD4^+$, $CD4^+/CD25^+$ and $CD11b^+/Gr-1^+$ cells in paw joint were significantly decreased compared with control. The absolute number of $B220^+/CD23^+$ and $MHCII^+/CD11c^+$ cell in draining lymph node were significantly decreased compared with control. The levels of IgG was decreased and The levels of IgM was significantly decreased compared with control. Anti-collagen II in serum was significantly decreased compared with control. With the hematoxylin and eosin stain, the cartilage destruction and synovial cell proliferation were decreased compared with control. With the Masson's trichrome stain, the expression of collagen fibers was decreased compared with control. Results showed that GJB-6 had immunomodulatory effects. So we expect that GJB-6 should be used as a effective drugs for not only rheumatoid arthritis but also another auto-immune disease.

Effect of Simvastatin collagen graft on wound healing of defective bone (Simvastatin이 골결손부 회복에 미치는 영향)

  • Kang, Jung-Ho;Kim, Gyu-Tae;Choi, Yong-Suk;Lee, Hyeon-Woo;Hwang, Eui-Hwan
    • Imaging Science in Dentistry
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    • v.38 no.3
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    • pp.133-146
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    • 2008
  • Purpose : To observe and evaluate the effects of Simvastatin-induced osteogenesis on the wound healing of defective bone. Materials and Methods : 64 defective bones were created in the parietal bone of 32 New Zealand White rabbits. The defects were grafted with collagen matrix carriers mixed with Simvastatin solution in the experimental group of 16 rabbits and with collagen matrix carriers mixed with water in the controlled group. The rabbits were terminated at an interval of 3, 5, 7, and 9 days, 2, 4, 6, and 8 weeks after the formation of defective bone. The wound healing was evaluated by soft X-ray radiography. The tissues within defective bones were evaluated through the analysis of flow cytometry for the manifestation of Runx2 and Osteocalcin, and observed histopathologically by using H-E stain and Masson's trichrome stain. Results : 1. In the experimental group, flow cytometry revealed more manifestation of Runx2 at 5, 7, and 9 days and Osteocalcin at 2 weeks than in the controlled groups, but there was few difference in comparison with the controlled group. 2. In the experimental group, flow cytometry revealed considerably more cells and erythrocytes at 5, 7, and 9 days in comparison with the controlled group. 3. In the experimental group, soft x-ray radiography revealed the extended formation of trabeculation at 2, 4, 6, and 8 weeks. 4. Histopathological features of the experimental group showed more fibroblasts and newly formed vessels at 5 and 7 days, and the formation of osteoid tissues at 9 days, and the newly formed trabeculations at 4 and 6 weeks. Conclusion : As the induced osteogenesis by Simvastatin, there was few contrast of the manifestation between Runx2 and Osteocalcin based on the differentiation of osteoblasts. But it was considered that the more formation of cells and erythrocytes depending on newly formed vessels in the experimental group obviously had an effect on the bone regeneration.

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The Preventive Effect of Allergic Inflammation by Induction of Oral Tolerance in a Mouse Model of Chronic Asthma (마우스 만성천식모델에서 경구면역관용 유도에 의한 알레르기 염증의 예방효과)

  • Kim, Jin Sook;Lee, Jung Mi;Kim, Seung Joon;Lee, Sook Young;Kwon, Soon Seog;Kim, Young Kyoon;Kim, Kwan Hyoung;Moon, Hwa Sik;Song, Jeong Sup;Park, Sung Hak
    • Tuberculosis and Respiratory Diseases
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    • v.57 no.5
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    • pp.425-433
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    • 2004
  • Background : Induction of oral tolerance (OT) has been known to prevent allergic inflammation in acute asthma model within 4 weeks. However it is remained whether induction of OT may effectively prevent allergic inflammation in chronic asthma model over 4 weeks. We observed the effect of induction of OT on allergic inflammation and airway remodeling in chronic asthma model up to 8 weeks. Methods : 5-week-old female BALB/c mice divided into 4 groups-control group, asthma group, low dose OT group, and high dose OT group. To induce oral tolerance mice were fed ovalbumin (OVA) before sensitization with OVA and aluminum hydroxide-1 mg for 6 consecutive days in the low dose OT group and 25 mg once in the high dose OT group. Mice in the asthma group were fed phosphate buffered saline instead of OVA. After sensitization followed by repeated challenge with aerosolized 1% OVA during 6 weeks, enhanced pause (Penh), inflammatory cells, IL-13, and IFN-${\gamma}$ levels in bronchoalveolar lavage (BAL) fluids as well as OVA-specific IgE, IgG1, and IgG2a levels in serum were measured. In addition the degree of goblet cell hyperplasia and peribronchial fibrosis were observed from lung tissues by PAS and Masson's trichrome stain. Results : Both OT groups showed a significant decrease in Penh, inflammatory cells, IL-13, and IFN-${\gamma}$ levels in BAL fluids as well as OVA-specific IgE, IgG1, and IgG2a levels in serum compared with the asthma group (P<0.05). In addition, the degree of goblet cell hyperplasia and peribronchial fibrosis were significantly attenuated in both OT groups compared with the asthma group (P<0.01). Conclusion : These results suggest that induction of OT may effectively prevent allergic inflammation as well as airway remodeling even in chronic asthma model up to 8 weeks.

Effects of Intramedullary Vascularized Muscle Flap in Regeneration of Lyophilized, Autografted Humeral Head in Rabbits (골수강내 혈관성 근피판 이식이 동결 건조후 자가 이식된 관절연골의 재생에 미치는 효과)

  • Rhee, Seung-Koo;Kim, Sung-Tae;Park, Jin-Il
    • 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.

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AN EXPERIMENTAL STUDY ON THE HEALING PROCESS AFTER THE IMPLANTATION OF VARIOUS BONE SUBSTITUTES IN THE RATS (백서에서 수종의 골대체재료 매식후 치유과정에 관한 실험적 연구)

  • Kim, Young-Kyun;Kim, Su-Gwan;Lee, Jun-Gil;Lee, Mi-Hyang;Cho, Jae-O
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.27 no.1
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    • pp.15-24
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    • 2001
  • The purpose of this study is to evaluate the tissue response in applying of various bone substitutes included toothash-plaster mixture, resorbable hydroxylapatite (HA) and demineralized freeze-dried bone and to show the clinical usefulness of toothash-plaster mixture for the repair of craniomaxillofacial bone defect. For this experiment, 100 Sprague-Dawley rats weighing 200gm or more were used. There were four experimental groups: group I, toothash-plaster mixture; group II, demineralized freeze-dried bone; group III, resorbable HA; and group IV, control group. A full thickness, round bone defect measuring 10mm in diameter was created in the midcranium, and the substitutes cited above were embedded in the experimental rats based on their group assignment. Blood clot was filled in the rats assigned to the control group. Experimental rats were sacrificed on the 1st, 3rd, 5th, 8th, 12th and 24th week after implantation and stained with the hematoxylineosin, Masson's Trichrome, using Van Gieson's stain method, and were examined under light microscope. The results were as follows: 1. In all the groups, prominent inflammatory reaction and the infiltration of multinucleated giant cells were noted during the early stage. Gradual healing decreased this reaction. 2. Among the rats in the experimental group II, which were given demineralized freeze-dried bone implants, active formation of new bone traveculae manifested. Chondroid tissues appeared, and it was suggested that the defect was filled with newly formed bone by virtue of osteoinductive activity. On the 12th week after the experiments, most of the defect was filled with newly formed bone trabeculae. 3. In experimental groups I and III, it was noted that HA manifested a healing process similar to that characterized by the toothash-plaster mixture, but inflammatory reaction was more prominent in experimental group I. Active osteoblasts were observed along the periphery of osteoid tissues, while newly formed bone trabeculae appeared adjacent to the implanted materials three weeks later. Formation increased to the extent that newly formed bone trabeculae fused directly with the host bone. Increase in new bone ingrowth into the filling materials was revealed by both experimental groups. 4. In the control group, new bone formation adjacent to the host bone was observed, but most of the defect was filled with mature connective tissue 24 weeks after the experiments.

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Effects of Intramedullary K-wire Insertion on Femoral Lengthening in Canine with Monolateral External Fixator (단측성 외고정 기구를 이용한 개의 대퇴골 신연에 있어서 골수강내 K-강선 삽입의 효과)

  • Hwang, Jae-Min;Kim, Young-Ki;Chin, Hee-Tec;Seok, Seong-Hoon;Lee, Hee-Chun;Koh, Phil-Ok;Chang, Hong-Hee;Lee, Hyo-Jong;Yeon, Seong-Chan
    • 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.