• Title/Summary/Keyword: histological evaluation

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Bone Healing in Ovariectomized-rabbit Calvarial Defect with Tricalcium Phosphate Coated with Recombinant Human Bone Morphogenetic Protein-2 Genetically Engineered in Escherichia coli

  • Kim, Jung-Han;Kim, Chang-Joo;Shin, Sang-Hun
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.36 no.2
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    • pp.37-49
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    • 2014
  • Purpose: This study compares the bone formation ability of tricalcium phosphate (TCP) with and without recombinant human bone morphogenetic protein-2 (rhBMP-2) and assesses TCP as a carrier of rhBMP-2. Methods: Bilateral round defects (diameter: 8.0 mm) were formed in the cranium of eight New Zealand white rabbits. The defects were grafted with TCP only (control group) or with rhBMP-2-coated TCP (experimental group). The animals were sacrificed at 1st week, 2nd week, 4th week, and 8th week postoperatively; two rabbits sacrificed each time. The skulls were harvested and subjected to radiographic and histological examination. Results: Radiologic evaluation showed faster bone remodeling in the experimental group than in the control group. Histologic evaluation (H&E, Masson's trichrome stain) showed rapid bone formation, remodeling and calcification in the 1st and 2nd week in the experimental group. Immunohistochemical evaluation showed higher expression rate of osteoprotegerin, receptor activator of nuclear factor ${\kappa}B$ ligand, and receptor activator of nuclear factor ${\kappa}B$ in the experimental group at the 1st and 2nd week than in the control group. Conclusion: rhBMP-2 coated TCP resulted in rapid bone formation, remodeling, and calcification due to rhBMP-2's osteogenic effect. TCP performed properly as a carrier for rhBMP-2. Thus, the use of an rhBMP-2 coating on TCP had a synergic effect on bone healing and, especially, bone remodeling and maturation.

Computer-aided design/computer-aided manufacturing of hydroxyapatite scaffolds for bone reconstruction in jawbone atrophy: a systematic review and case report

  • Garagiola, Umberto;Grigolato, Roberto;Soldo, Rossano;Bacchini, Marco;Bassi, Gianluca;Roncucci, Rachele;De Nardi, Sandro
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.38
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    • pp.2.1-2.9
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    • 2016
  • Background: We reviewed the biological and mechanical properties of porous hydroxyapatite (HA) compared to other synthetic materials. Computer-aided design/computer-aided manufacturing (CAD/CAM) was also evaluated to estimate its efficacy with clinical and radiological assessments. Method: A systematic search of the electronic literature database of the National Library of Medicine (PubMed-MEDLINE) was performed for articles published in English between January 1985 and September 2013. The inclusion criteria were (1) histological evaluation of the biocompatibility and osteoconductivity of porous HA in vivo and in vitro, (2) evaluation of the mechanical properties of HA in relation to its porosity, (3) comparison of the biological and mechanical properties between several biomaterials, and (4) clinical and radiological evaluation of the precision of CAD/CAM techniques. Results: HA had excellent osteoconductivity and biocompatibility in vitro and in vivo compared to other biomaterials. HA grafts are suitable for milling and finishing, depending on the design. In computed tomography, porous HA is a more resorbable and more osteoconductive material than dense HA; however, its strength decreases exponentially with an increase in porosity. Conclusions: Mechanical tests showed that HA scaffolds with pore diameters ranging from 400 to $1200{\mu}m$ had compressive moduli and strength within the range of the human craniofacial trabecular bone. In conclusion, using CAD/CAM techniques for preparing HA scaffolds may increase graft stability and reduce surgical operating time.

The Effect of Fibrillar Collagen on Bony Healing of Calvarial Defect in Rats (골 조직 치유과정에서 Collagen 막의 효과)

  • Kim, Jae-Bung;Lee, Jae-Mok;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.29 no.2
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    • pp.355-373
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    • 1999
  • Many researches have been reported that collagen as cellular stroma, matrix of grafting materials, mediator of agents for the purpose of promoting healing process invivo, but the responses in vivo were seen various. The goal of this experiment is to assess the effect of collagen on bony healing, through histological evaluation of implanted collagen on the calvarial defect in rats. 2-month-old Sprague-Dawley, 24 rats were used and 12 rats assigned to each group of control and test. Defect of 5mm in diameter was made on the calvarial bone with trephine bur. Following thorough saline rinse, defect of control group was left in empty and that of experimental group was filled with fibrillar collagen($COLLATAPE^{(R)}$, COLLA-TEC. INC. U.S.A.) soaked in saline. 3 rats in each group were sacrificed at 3, 7, 14, 21 days after operation respectively, and the tissue blocks were prepared for light microscope with H-E for evaluation of overall healing, with TRAP(tartrate resistant acid phosphatase) for evaluation of osteoclastic activity and with immunohistochemical staining for macrophages. The results were as follows : 1. In the control group, inflammatory responses were disappeared at day 14, but, in the experimental group inflammatory infiltrates were reduced at day 21. Thus, the experimental group showed more severe soft tissue inflammation than control group. 2. Both control and experimental group showed slight appositional growth at day 7 and gradual bony growth to 21th day. But, complete bony healing of the defect was not shown. There was no significant difference in bony healing between control and experimental group 3. Specific response of macrophages for implanted collagen was observed at day 14 in the experimental group. In conclusion, although fibrillar collagen caused inflammation of soft tissue during initial healing period, inflammatory responses by fibrillar collagen didn't inhibit bony regeneration and implanted collagen was biodegradaded by macrophages. Thus, we expect that fibrillar collagen can be used for useful mediator of graft materials or growth factors.

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Study of bone healing pattern in extraction socket after application of demineralized dentin matrix material (자가치아 뼈 이식재의 발치와내 이식 후 골 치유 양상에 관한 연구)

  • Chung, Jae-Ho;Lee, Jae-Hoon
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.37 no.5
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    • pp.365-374
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    • 2011
  • Introduction: Research on dental bone graft material has been actively conducted. Recently, demineralized dentin matrix material has been developed and introduced. This study examined the effect of demineralized dentin matrix material on bone healing. Subjects and Methods: The patients who received no treatment after extraction were used as the control group and patients who underwent demineralized dentin matrix material application in the extraction socket after extraction were used as the experimental group. Panorama radiography was performed at the baseline and at 3.5 months after graft material placement and CT was taken at 3.5 months after graft material placement for a radiologic evaluation. Bony tissue specimens were collected from the alveolar crest in the middle of the extraction socket using a 2 mm trephine bur after 3.5 months for the histology and hostomorphometric study. Results: 1. On the panoramic view, a higher bone density was observed in the subject group. 2. On the panoramic view, the bone density increased significantly in the extraction socket, from the baseline to 3.5 months: a 7 and 10 gray-level scale was observed in the control and experimental group, respectively (P<0.05). 3. The CT view evaluation at 3.5 months revealed significantly higher bone density in the subject group than the control group (P<0.05). 4. The histological findings showed more active new bone and lamellar bone formation in the subject group. Dentin with osteoinduction ability and enamel with osteoconduction ability appeared. 5. On histomorphometric analysis, the subject group showed significantly more new bone, lamellar bone area and lower soft tissue area (P<0.05). The difference between the groups was significant (P<0.05). Conclusion: Bone healing was improved after the application of demineralized dentin matrix material and there was active new bone and lamellar bone formation.

Guided bone regeneration using demineralized allogenic bone matrix with calcium sulfate: case series

  • Kim, Young-Kyun;Lee, Ji-Young;Kim, Su-Gwan;Lim, Seung-Chul
    • The Journal of Advanced Prosthodontics
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    • v.5 no.2
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    • pp.167-171
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    • 2013
  • PURPOSE. The purpose of this case series was to evaluate the effect of guided bone regeneration using demineralized allogenic bone matrix with calcium sulfate. MATERIALS AND METHODS. Guided bone regeneration using Demineralized Allogenic Bone Matrix with Calcium Sulfate ($AlloMatrix^{TM}$, Wright. USA) was performed at the time of implant placement from February 2010 to April 2010. At the time of the second surgery, clinical evaluation of bone healing and histologic evaluation were performed. The study included 10 patients, and 23 implants were placed. The extent of bony defects around implants was determined by measuring the horizontal and vertical bone defects using a periodontal probe from the mesial, distal, buccal, and lingual sides and calculating the mean and standard deviation of these measurements. Wedge-shaped tissue samples were obtained from 3 patients and histologic examination was performed. RESULTS. In clinical evaluation, it was observed that horizontal bone defects were completely healed with new bones, and in the vertical bone defect area, 15.1% of the original defect area remained. In 3 patients, histological tests were performed, and 16.7-41.7% new bone formation was confirmed. Bone graft materials slowly underwent resorption over time. CONCLUSION. $AlloMatrix^{TM}$ is an allograft material that can be readily manipulated. It does not require the use of barrier membranes, and good bone regeneration can be achieved with time.

Evaluation of Bilayer Polycaprolactone Scaffold for Osteochondral Regeneration in Rabbits

  • Park, Min-hyeok;Hwang, Ya-won;Jeong, Do-Sun;Kim, Gon-hyung
    • Journal of Veterinary Clinics
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    • v.33 no.6
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    • pp.332-339
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    • 2016
  • Polycaprolactone (PCL) scaffold have been developed as an alternative to natural donor tissue to repair a large osteochondral defect. The objective of this study is to evaluate efficacy and biocompatibility of bilayer PCL scaffold implanted for osteochondral repair in rabbit. Twenty-two male New Zealand White rabbits were used in this animal experiment. Rabbits were divided into three groups. Experimental surgery was carried out under general anesthesia. Osteochondral defects (5 mm diameter and 5 mm deep) were made in the center of the patellar groove using a 5 mm diameter biopsy punch. In group I (3D plotting) and group II (salt-leaching), the scaffold was implanted using the press-fitted technique into the defect. In control group, after osteochondral defect was created, the defect was left without implant. After four and eight weeks, rabbits were sacrificed and the defects were evaluated by macro -and microscopical methods. There were not found animal death and severe inflammatory evidence during the experimental periods. There were no significant differences between the experimental groups in gross evaluation. However the group I scored significantly higher than group II at 8 weeks in histological evaluation (P < 0.05). The 3-D plotting PCL scaffold was more suitable method for reconstruction of osteochondral defect than a salt-leaching PCL scaffold.

Chronic HBV Infection in Children: The histopathologic classification and its correlation with clinical findings (소아의 만성 B형 간염: 새로운 병리조직학적 분류와 임상 소견의 상관 분석)

  • Lee, Seon-Young;Ko, Jae-Sung;Kim, Chong-Jai;Jang, Ja-June;Seo, Jeong-Kee
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.1 no.1
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    • pp.56-78
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    • 1998
  • Objective: Chronic hepatitis B infection (CHB) occurs in 6% to 10% of population in Korea. In ethinic communities where prevalence of chronic infection is high such as Korea, transmission of hepatitis B infection is either vertical (ie, by perinatal infection) or by close family contact (usually from mothers or siblings) during the first 5 years of life. The development of chronic hepatitis B infection is increasingly more common the earlier a person is exposed to the virus, particularly in fetal and neonatal life. And it progress to cirrhosis and hepatocellular carcinoma, especially in severe liver damage and perinatal infection. Histopathology of CHB is important when evaluating the final outcomes. A numerical scoring system which is a semiquantitatively assessed objective reproducible classification of chronic viral hepatitis, is a valuable tool for statistical analysis when predicting the outcome and evaluating antiviral and other therapies. In this study, a numerical scoring system (Ludwig system) was applied and compared with the conventional histological classification of De Groute. And the comparative analysis of cinical findings, family history, serology, and liver function test by histopathological findings in chronic hepatitis B of children was done. Methods: Ninety nine patients [mean age=9 years (range=17 months to 16 years)] with clinical, biochemical, serological and histological patterns of chronic HBV infection included in this study. Five of these children had hepatocelluar carcinoma. They were 83 male and 16 female children. They all underwent liver biopsies and histologic evaluation was performed by one pathologist. The biopsy specimens were classified, according to the standard criteria of De Groute as follows: normal, chronic lobular hepatitis (CLH), chronic persistent hepatitis (CPH), mild to severe chronic active hepatitis (CAH), or active cirrhosis, inactive cirrhosis, hepatocellular carcinoma (HCC). And the biopsy specimens were also assessed and scored semiquantitatively by the numerical scoring Ludwig system. Serum HBsAg, anti-HBs, HBeAg, anti-HBe, anti-HBc (IgG, IgM), and HDV were measured by radioimunoassays. Results: Male predominated in a proportion of 5.2:1 for all patients. Of 99 patients, 2 cases had normal, 2 cases had CLH, 22 cases had CPH, 40 cases had mild CAH, 19 cases had moderate CAH, 1 case had severe CAH, 7 cases had active cirrhosis, 1 case had inactive cirrhosis, and 5 cases had HCC. The mean age, sex distribution, symptoms, signs, and family history did not differ statistically among the different histologic groups. The numerical scoring system was correlated well with the conventional histological classification. The histological activity evaluated by both the conventional classification and the scoring system was more severe as the levels of serum aminotransferases were higher. In contrast, the levels of serum aminotransferases were not useful for predicting the degree of histologic activity because of its wide range overlapping. When the histological activity was more severe and especially the cirrhosis more progressing, the prothrombin time was more prolonged. The histological severity was inversely related with the duration of seroconversion of HBeAg. Conclusions: The histological activity could not be accurately predicted by clinical and biochemical findings, but by the proper histological classification of the numerical scoring system for the biopsy specimen. The numerical scoring system was correlated well with the conventional histological classification, and it seems to be a valuable tool for the statistical analysis when predicting the outcome and evaluating effects of antiviral and other therapies in chronic hepatitis B in children.

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Biochemical and Nutritional Effects of Rice, Barley, and Millet on Rats (백미대맥(白米大麥) 및 속(粟) 사료(飼料)가 실험용(實驗用) 백서(白鼠)에 미치는 영양학적(營養學的) 및 생물화학적(生物化學的) 연구(硏究))

  • Haw, Kum
    • Journal of Nutrition and Health
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    • v.1 no.1
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    • pp.9-18
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    • 1968
  • Dividing into six(6) experimental groups; Stock Diet(SD) Group, Rice Diet(RD) Group, Rice Diet supplemented with Barley(RD-BD) Group, Barley Diet(BD) Group, Rice Diet supplemented with Millet(RD-MD) Group, and Millet Diet(MD) Group, biochemical and nutritional effects of rice, barley, and millet on rats are studied. The grains used in this experiment are polished ones and purchased from market. Six(6) weaning male albino rats are housed into the individual cages in each group and fed with each experimental diet and water ad libitum for 21 weeks. The composition of the experimental diets are shown in Table 1. At the end of experiment, the appearences of rats are observed and the bloods are sampled by heart puncture method following ether anesthesia for determinations of serum protein and serum cholesterol. And the livers are sampled immediately right after heart after heart puncture for determinations of liver weight, liver nitrogen, liver fat, and liver glycogen. And also liver tissues are observed histologically. The result of growth experiment is shown in Table 2 and Figure 7. The results of blood analysis are shown in Table 3 and the results of liver analysis are shown in Table 4. And the histological observations of liver tissue are shown in Figures 1, 2, 3, 4, 5, and 6. According to the above results, it could be summarized as follows; 1) The normal appearences are observed at SD Group and BD Group. Alopecia is observed at RD, RD-­BD, RD-MD, and MD groups and the wet appearences of hair are observed at RD-MD and MD groups. The color of hair is changed to yellowish white at RD, RD-MD groups. The maximum growth inhibition, is observed at MD group and the growths of RD, RD-BD, BD, and RD-MD groups are significantly lower than that of SD group. The growth of RD group is lower than those of RD-BD and BD groups, though this difference is not significant. 2) At the blood analysis, the highest serum protein content is shown at SD group and the lowest serum protein is observed at MD group. The serum cholesterol contents are not significantly different among the six(6) experimental groups. 3) The highest liver weight is observed at SD group and the lowest weight is shown at MD group. However, oppositely the highest liver weight per 100 grams of body weight at MD group and the lowest liver weight per 100 grams of body weight at SD group are observed as shown in Table 6. 4) The differences of liver nitrogen contents of RD, RB-BD, BD, RD-MD, and MD Groups are not significant except SD group showing the highest content. 5) The significant highest content of liver fat is observed at RD group and the lowest content of liver fat is shown at SD group. And no significant differences in contents of liver glycogen are observed among the six(6) experimental groups. 6) No significant abnormal histological changes of liver tissue are observed at SD, RD-BD, and BD groups. However, the fat metamorphosis at peripheral zone of liver tissue is observed at RD Group and the some histological abnormal changes are observed at RD-MD and MD groups. According to the above summaries, the followings might be estimated and recommended; 1) The BD group shows histological normality of liver tissue, improvement of liver fat and liver nitrogen, normality of appearence, and better growth compared with RD Group. This could be indicated that barley is nutritionally and biochemically better than rice as the feeding diet on rat. And it might be considered that there might be the species speciality for nutritionary evaluation of food. 2) Recently, Marshall indicated that the high consumption of carbohydrate causes fatty liver and Kondo reported that the higher consumption of rice causes the more heart disease and the shorter life span. Considering these reports, the higher consumption of rice as a main dish in our country should be considered as an important national nutritional problem. And this might be improved by supplementing with barley or other grains to rice.

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3-Dimensional Micro-Computed Tomography Study on Bone Regeneration with Silk Fibroin, rh-Bone Morphogenetic Protein Loaded-Silk Fibroin and Tricalcium Phosphate Coated-Silk Fibroin in Rat Calvaria Defect

  • Pang, Eun-O;Park, Young-Ju;Park, Su-Hyun;Kang, Eung-Sun;Kweon, Hae-Yong;Kim, Soeng-Gon;Ko, Chang-Yong;Kim, Han-Sung;Nam, Jeong-Hun;Ahn, Jang-Hun;Chun, Ji-Hyun;Lee, Byeong-Min
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.34 no.1
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    • pp.1-11
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    • 2012
  • Purpose: The purpose of this study was to evaluate the bone regeneration capacity of silk fibroin (SF) when combined with beta tricalcium phosphate (${\beta}$-tricalcium phosphate [TCP]) and rh-bone morphogenetic protein (BMP) in vivo by micro-computed tomography (CT), soft x-ray, and histological analysis. Methods: A total of 56 critical size defects formed by a trephine bur made on 28 adult female Spague-Dawley rats were used for this study and the defect size was 5.0 mm in diameter. The defects were transplanted with (1) no graft material (raw defect), (2) autogenous bone, (3) SF ($10{\mu}g$), (4) SF-BMP ($10{\mu}g$, $0.8{\mu}g$ each), and (5) SF+${\beta}$-TCP ($10{\mu}g$). At 4 and 8 weeks after operation, the experimental animals were sacrificed. Samples were evaluated with soft x-ray, histological examinations and 3-dimensional micro-CT analysis. Results: In the 3-dimensional micro-CT evaluation, bone volume and bone surface data were higher in the SF-BMP ($12.8{\pm}1.5$, $138.6{\pm}45.0$ each) (P<0.05) and SF-TCP ($12.3{\pm}1.5$, $144.9{\pm}30.9$ each) group than in the SF group ($6.1{\pm}3.3$, $77.2{\pm}37.3$ each) (P<0.05), except for the autogenous group ($15.0{\pm}3.0$, $190.7{\pm}41.4$ each) at 4 weeks. At 8 weeks, SF-BMP ($16.8{\pm}3.5$, $173.9{\pm}34.2$ each) still revealed higher (P<0.05) bone volum and surface, but SF-TCP ($11.3{\pm}1.5$, $1132.9{\pm}52.1$ each) (P=0.5, P=0.2) revealed the same or lower amount compared with the SF group ($13.8{\pm}2.7$, $127.5{\pm}44.8$ each). The % of bone area determined by radiodensity was higher in the SF-TCP ($31.4{\pm}9.1%$) and SF-BMP ($36.2{\pm}16.2%$) groups than in the SF ($19.0{\pm}10.4$) group at the period of 4 weeks. Also, in the histological evaluation, the SF-BMP group revealed lower inflammation reaction, lower foreign body reaction and higher bone healing than the SF group at postoperative 4 weeks and 8 weeks. The SF-TCP group revealed lower inflammation at 4 weeks, but accordingly, as the TCP membrane was absorbed, inflammatory and foreign body reaction are increased at 8 weeks. Conclusion: The current study provides evidence that the silk fibrin can be used as an effective grafted material for tissue engineering bone generation through a combination of growth factor or surface treatment.

Evaluation of Neoadjuvant Chemotherapy Effect in Osteosarcoma (골육종에서 술전 항암화학요법의 효과 판정)

  • Joo, Min Wook;Kang, Yong-Koo;Yoo, Ie Ryung;Choi, Woo Hee;Chung, Yang-Guk;Kim, Dong-Hyun;Kang, Jin-Woo
    • The Journal of the Korean bone and joint tumor society
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    • v.20 no.2
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    • pp.66-73
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    • 2014
  • Purpose: Various diagnostic imaging modalities have been used to evaluate the effect of neoadjuvant chemotherapy for osteosarcoma early and noninvasively. We evaluated the effectiveness of imaging studies of plain radiographs and positron-emission tomography/computed tomography (PET/CT) in predicting neoadjuvant chemotherapy effect for osteosarcoma and tried to establish a general principle in interpretation of PET/CT parameters. Materials and Methods: Eighteen patients who underwent two cycles of neoadjuvant chemotherapy and surgical excision for osteosarcoma were enrolled. There were 13 males and 5 females, with a median age of 19 (11-63) years. Fifteen patients of 18 had the American Joint Committe on Cancer (AJCC) stage IIB. They had plain radiographs and PET/CT before and after neoadjuvant chemotherapy. The resected tumor specimens were pathologically examined to determine histological response grade using a conventional mapping method. Statistical analysis was performed to evaluate the correlation between histopathological necrosis rate, and radiographic finding category, post-chemotherapy maximum standardized uptake value (SUVmax), average standardized uptake value and metabolic tumor volume (MTV) as well as reduction rates of them. Results: Eight patients were good responders to neoadjuvant chemotherapy based on histological evaluation. Median SUVmax reduction rate was 73 (23-77) % in good responders and 42 (-32-76) % in poor responders. Median MTV reduction rate was 93.5 (62-99) % in good responders and 46 (-81-100) % in poor responders. While radiographic finding category was not different according to histological response (p=1.0), SUVmax reduction rate was significantly different (p=0.041). Difference in MTV reduction rates approached statistical significance as well (p=0.071). Conclusion: While radiographic finding category was not reliable to assess neoadjuvant chemotherapy effect for osteosarcoma, reduction rate of SUVmax was a useful indicator in this study. As parameters of PET/CT can be influenced by various factors of settings, different centers have to make an effort to establish their own standard of judgement with reference of previous studies.