• Title/Summary/Keyword: rat

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Induction of Hepatic Microsomal Cytochrome P450 by N,N-dimethylformamide in Sprague-Dawley Rats (흰쥐에서 N,N-dimethylformamide에 의한 간장의 Microsomal Cytochrome P450의 유도)

  • Koh, Sang-Baek;Cha, Bong-Suk;Kang, Seung-Kyu;Joung, Hyo-Seok;Kim, Ki-Woong
    • Journal of Preventive Medicine and Public Health
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    • v.32 no.1
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    • pp.88-94
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    • 1999
  • Objectives. In order to gain a better understanding of the mechanism of DMF toxicity, recent studies have focused on hepatic drug metabolizing enzymes. In this study, we investigated the effects of DMF on the induction of P450 and the activities of other related enzymes in rat liver microsomes. Methods. DMF was administered to male Sprague Daweley rats by intraperitoneal injection at 0(control), 450(D1), 900(D2), 1,800(D3) mg DMF/kg body weight in olive oil once a day for three days. Hepatic P450 was measured by method of Omura and Sato. We evaluated selective assays for the three drug metabolizing cytochrome P450 isoenzymes 1A1, 2B1 and 2E1. Results. The content of microsomal protein, P450 and b5 were tended to be decreased in DMF treated group, but they were not statistically significant. The activity of NADPH-cytochrome P450 reductase was significantly increased dose dependently(p<0.01), but the activity of NADH-b5 reductase was decreased in the treated group(p<0.01). The activities of PROD and EROD were not significant between control and treated group. The activities of pNPH in the DMF treated groups were higher than that of the control group(p<0.01). When Western immunoblottings were carried out utilizing three monoclonal antibodies which were specific against P4501A1/1, P4502B1/2 and P4502E1, the strong density band corresponding to P4502E1 was observed with the microsomes obtained from the rats treated with DMF. But there were no significant increased in the P4501A1/2 and P4502B1/2 band densities in immunoblotting. Conclusions. These result suggested that P4502E1 was inducible by DMF and P4502E1 isozyme might be responsible for the hydroxylation of DMF to HMMF.

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On the Biological Functions of Equine Chorionic Gonadotropin (말의 융모성 성선자극 호르몬의 생화학적 기능)

  • 민관식;윤종택
    • Korean Journal of Animal Reproduction
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    • v.26 no.3
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    • pp.299-308
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    • 2002
  • In horse, a single gene encodes both eCG and eLH $\beta$ subunits. The difference between eCG and eLH lies in the structure of their glycoresidues, which are both sialylated and sulfated in LH and sialylated in CG eCG consists of highly glycosyiated $\alpha$- and $\beta$-subunits and is an unique member of the gonadotropin family because it elicits response characteristics of both FSH and LH in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of gonadotropin structure-function relationships and the understanding of the molecular bases of the specific interactions of these hormones with their receptors. Thus, eCG is a dintinct molecule from the view points of its biological function and glycoresidue structures. The oligosaccharide at Asn 56 of the $\alpha$-subunit plays an indispensable role, whereas the carboxyl-terminal extension of the eCG $\beta$-subunit with its associated O-linked oligosaccharides is not improtant for, the in vitro LH-like activity of eCG. In contrast, both N- and O-linked oligosaccharides play important roles for FSH-like activity and increase FSH-like activity by removal of N- and O-linked oligosaccharides. Therefore, the dual LH- and FSH-like activities of eCG can be clearly separated by removal of either the N-linked oligosaccharide on the $\alpha$-subunit or CTP-associated O-linked oligosaccharides from its $\beta$-subunit. The glycoresidues seem to play crucial roles fer biological activities. The tethered-eCG was effciently secreted and showed similar LH-like activity to the dimeric eCG $\alpha$/ $\beta$ and native eCG. FSH-like activity of the tethered-eCG was also shown similarly in comparison with the native and wild type eCG $\alpha$/ $\beta$. Our data for the first time suggest that the tethered-eCG can be expressed efficiently and the produced product by the CHO-Kl cells is fully LH- and FSH-like activities in rat in vitro bioassay system. Our results also suggest that this molecular can imply particular models ot FSH-like activity not LH-like activity in the eCG. Taken together, these data indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion.

Effects of Hagocho (Prunella vulgaris L.), Gamgook (Chrysanthemum indicum L.) and Galgeun (Pueraria Radix) on Plasma Lipid Compositin and Histological Consideration in Hyperlipidemic Rat (하고초, 감국 및 갈근 증류액이 비만쥐의 지질구성 및 각종 장기의 해부조직학적 소견에 미치는 영향)

  • Park, Chan-Hee;Kim, Nam-Young;Nam, Eun-Jeung;Kim, Sang-Hyeun;Lee, Ji-Hyang;Lee, Eun
    • Korean Journal of Plant Resources
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    • v.20 no.1
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    • pp.22-27
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    • 2007
  • Effects of prunella vulgaris L., chrysanthemum indicum L. and pueraria Radix on plasma lipid composition and histological consideration were investigated in hyperlipidemic rats. Concentration of plasma ${\beta},-lipoprotein$ showed a tendence to decrease in prunella vulgaris L., chrysanthemum indicum L. and pueraria Radix treatment groups. However these values were showed not significantly different from control group. Concentration of plasma FFA in prunella vulgaris L., chrysanthemum indicum L. and pueraria Radix treatment groups showed a lower values compared to control group and concentration of plasma FFA of prunella vulgaris L. and pueraria Radix treatment group showed a lower values than prunella vulgaris L. and chrysanthemum indicum L. treatment group. Concentration of plasma glucose and triglyceride showed a tendence to decrease in prunella vulgaris L., chrysanthemum indicum L. and pueraria Radix treatment groups. However plasma glucose values showed not significantly different from control group. Plasma total cholesterol concentration showed a low and HDL-cholesterol concentration showed a high in prunella vulgaris L. and pueraria Radix treatment group. However LDL-cholesterol concentrations were not significantly different in treatment groups. Histological consideration of heart, liver and kidney in prunella vulgaris L. and pueraria Radix treatment groups showed a good features in fat accumulation condition than those of other treatment groups. However in the condition of high fat accumulation in tissues, heart, liver and kidney were showed a slight congestion and a bashed cell nucleus.

Nutritional Evaluation of Tofu Containing Dried Soymilk Residue(DSR) 1. Evaluation of Protein Quality (건조비지 첨가 두부의 영양적 품질평가 1. 단백질의 영양가)

  • Kweon, Mi-Na;Ryu, Hong-Soo;Moon, Jeung-Hye
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.3
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    • pp.255-261
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    • 1993
  • The effect of dried soymilk residue (DSR) on protein quality of tofu was studied. The amount of added DSR into soybean water extract was corresponding 10% (dry basis) of soybean used in tofu manufacturing. Proximate composition and in vitro protein qualities of soybeans at different stages of the conversion into tofu have also been investigated. Partially substituted tofu with DSR (TDSR) had higher moisture content (80.6%) than that of tofu prepared in traditional manner (TT). TDSR contained lower content of protein (38.9%) and total lipid (26.9%) compared to 45.8% of protein and 34.3% of total lipid for TT. A large amount of trypsin inhibitor (TI) in raw soybeans was diminished and extracted through tofu processing, and only 10~13% of TI in raw soybean remained in both tofu products (TDSR and TT). There was not a considerable difference in amino acid profiles between TT and TDSR, but TDSR had a higher content of lysine than that in TT. in vitro studies showed that TDSR and TT were comparable in terms of both in vitro digestibilities (90% over for four-enzyme digestibility and predicted digestibility) and discriminant computed protein efficiency ratio (2.07~2.14, DC-PER). Unlike those in vitro indices for protein quality, computed protein efficiency ratio (C-PER) of TDSR was much lower (1.4) than that of TT (1.95). It was revealed that C-PERs of tofu products were not in agreement with rat-PERs (1.7~1.9) in previous reports except for TT. However, DC-PER assay was more recommendable for protein quality of tofu products than C-PER assay.

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Effects of Pueraia mirifica on the Experimental Hepatocarcinogenesis in Rats and Gap Junctional Intercellular Communication (랫드의 실험적 간암 발생과정과 Gap Junction을 통한 세포간 정보전달에서 Pueraia mirfica의 효과)

  • 강경선;김경배;이재해;조성대;조종호;박준석;안남식;양세란;정지원
    • Journal of Food Hygiene and Safety
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    • v.16 no.3
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    • pp.212-220
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    • 2001
  • To investigate the modifying effect of Kwao Kreu, Pueraria mirifica (PM), we performed two kind of studies which are the non-surgical medium-term carcinogenicity study and the modulation of gap junctional intercellular communication study. The first study, a non-surgical medium-term carcinogenicity bioassay was done to investigate the modifying effect of Kwao Keru, Pueyaria mirifca (PH), a rejuvenating folk medicine from Thailand, on the male F344 rat liver. Specific pathogen free, male 6-week-old F3444 rats were divided into ten groups. To induce hepatocarcinogenesis, those in all groups were given a single i.p. injection of DEN (200 mg/kg) and were received two i.p. injection of DGA (300 mg/kg) at the ends of weeks 2 and 5. Rats of group 3-6 were given sodium phenobarbital (PB 0.05% in drink). A diet containing 10 mg/kg PM was given to group 2 during the post-initiation phase and to groups 4 and 5 during promotion and initiation phase, respectively. Group 6 was given the experimental diet alone throughout the experiment (8 weeks). Rats of group 7, 8, 9 and 10 were fed 1000 mg/kg PH in the same manner as group 2, 4, 5 and 6. All animals were sacrificed at 8 weeks after DEN administration. Result of the immunohistochemical staining of the glutathione S-transferase placental form (GST-p) indicated that the numbers and areas of the preneoplastic leisions were not significantly changed in all PM treatment group comparing to control group. Also the numbers and areas of GST-p positive foci among group 7, 8, 9 and 10 were not significantly changed in comparing to control group. To study the effect of PM on the modulation of gap junctional intercellular communication, the present study was performed scrape-loading dye transfer (SL/DT) assay in human keratinocytes. The results showed that PM could not modulate GJIC. These results indicate that Pueraria mirifica may have no carcinogenic effects on experimental hepatocarcinogenesis in rats and gap junctional intercellular communication in human keratinocyte.

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The Antioxidative Activity of Glutathione-Enriched Extract from Saccharomyces cerevisiae FF-8 in In Vitro Model System (In Vitro 과산화지질에 미치는 glutathione 고함유 효모 Saccharomyces cerevisiae FF-8의 항산화효과)

  • Lee Chi-Hyeoung;Cha Jae-Young;Jun Bang-Sil;Lee Ho-Jun;Lee Young-Chun;Cho Yong-Lark;Cho Young-Su
    • Journal of Life Science
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    • v.15 no.5 s.72
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    • pp.819-825
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    • 2005
  • The Antioxidative accvities of the cell free extracts containing high glutathione by Saccharomyces cerevisiae FF-8 were tested in vitro experimental models : DPPH method for radical scavenging activity, ferric TBA method and ferric thiocyanate method using linoleic acid and tissue microsome for lipid peroxidation inhibitions. The concentration of intercellular glutathione by cultivating S. cerevisiae FF-8 in the YM optimal medium obtained $204\mug/ml$, which was increased by 2.76-fold from $74\mug/ml$ in the YM basal medium. A comparition between the YM basal medium and the YM optimal medium on antioxidative substance produced by S. cerevisiae FF-8 was investigated. In DPPH ($\alpha, \alpha-diphenyl-\beta-picrylhydrazyl$) method, the electron donating activity of the glutathione produced by S. cerevisiae FF-8 cultured in the YM optimal medium was as high as that of BHT ($ 0.05\%w/v $). The antioxidative a.tivity was measured by inhibition against lipid peroxidation of rat tissues' microsomes. The results of anti-oxidant activity of the cell free extracts by S. rerevisiae FF-8 cultured in the YM optimal medium was shown in the following order . $ liver 60.98\% > kidney 56.43\% > heart 52.91\% > brain 52.13\% > testis 45.57\% > spleen 42.95\% $. In antioxidative activities determined by ferric thiocyanate method and TBA methods against lipid peroxidation, the lipid peroxidation in the control mixture increased more rapidly than the typical peroxidation curve of linoleic acid from one day. The antioxidative activity of the cell free extracts by cultivating S. cerevisine FF-8 in the YM optimal medium were higher than that of the YM basal medium. These data indicate that the cell free extracts containing a high intercellular glutathione of S. cerevisiae FF-8 cultured in YM optimal medium showed strong antioxidative capacities by DPPH radical scavenging activity and ferric thiocyanate and TBARS measurements.

Effects of a Combined Diet of Jerusalem Artichoke's Inulin, Lotus Leaf and Herb Extracts in Obesity-induced White Rat with Fat Diet (돼지감자의 이눌린, 연잎, 허브의 병합식이가 고지방식이로 유도된 비만흰쥐에 미치는 영향)

  • Lee, Eun-Hye;Lee, Ye-Jin;Choi, Ok-Byung;Kang, Sang-Mo
    • Applied Biological Chemistry
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    • v.50 no.4
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    • pp.295-303
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    • 2007
  • A preliminary diet experiment utilizing Jerusalem artichoke's inulin, lotus leaf powder, nettle powder and eucalyptus powder extract indicated that combining all four elements gave the most effective result. Therefore, a study was conducted to evaluate the effectiveness of combined diet for weight loss. In this study, Sprague-Dawley, male white rats about 200 g in weight was fed with high fat diet for 8 weeks in order to induce obesity followed by 4 week administration of combined diet to look into the effect of the diet. After a total of 12 weeks of feeding, factors relevant to weight, blood, and lipid metabolism by liver in the body were researched and histologic change was examined with optical microscope. In terms of weight change, both high fat diet group and regular diet group gained weight from high fat diet for 8 weeks compared to normal group. Then, for another 4 weeks, while normal group and high fat diet group kept gaining weight, combined diet group which was provided with high fat diet for 8 weeks, lost weight to the normal group level after 3 week administration of diet. However, after the 4th week of administration, the group weighted significantly less than the normal group and the efficiency of diet also significantly dropped. In the biochemical analysis of blood, total cholesterol, LDL-cholesterol, triglyceride, glucose, GOT, GPT, ${\gamma}-GTP$ and creatine showed significant increase in high fat diet group and there was no significant difference between diet group and normal group except for GPT, ${\gamma}-GTP$ and creatine. In the biochemical analysis of liver, there was significant increase in LDL-cholesterol, triglyceride of high fat diet group compared to normal group, while there was no significant difference in term of total cholesterol and HDL-cholesterol. Compared to normal group, diet group had higher HDL-cholesterol, while total cholesterol dropped significantly. There was no significant difference in terms of LDL-cholesterol and triglyceride. Besides, in high fat diet group, observation of histologic change in liver and change in ultrastructure showed volume increase of hepatic cell and severe fatty degeneration in hepatic cell around hepatic vein. However in diet group, like normal group, no pathological change was observed in terms of cytoplasm, nucleus and capillary in hepatocyte and the alignment of hepatocyte had regularity thanks to the administration of combined diet. Therefore, combined diet utilizing Jerusalem artichoke's inulin, lotus leaf powder, nettle powder and eucalyptus powder was proven to be an effective measure to prevent and improve obesity as a result of abnormal adipose deposition.

Effect of Korean Mistletoe Extract and Lectin on the Preneoplastic Hepatic Lesion and Apoptosis in Experimental Hepatocarcinogenesis (실험적 간암모델에서 한국산 겨우살이 추출물 및 렉틴 투여가 전암성 병변의 생성 및 Apoptosis에 미치는 영향)

  • 김미정;김정희;이미숙
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.5
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    • pp.782-787
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    • 2002
  • This study was done to investigate the effects of Korean mistletoe water extract and lectin on the apoptosis and preneoplastic lesion in chemically induced rat hepatocarcinogenesis. To attain the above objectives, weanling Sprague-Dawley male rats were fed modified AIN-76 diets containing 10% corn oil for 9 weeks. One week after feeding starts, rats were intraperitoneally injected twice with a dose of diethylnitrosamine (DEN, 50 mg/kg body weight (BW). Rats were provided with 0.05% phenobarbital (PB) in drinking water from one week after DEN treatment until the end of experiment. During the period of PB treatment, rats were injected with mistletoe extract (100 $\mu\textrm{g}$/kg BW) and lectin (10 $\mu\textrm{g}$/kg BW) twice a week. At the end of 9th week, rats were sacrificed and the formation of hepatic glutathione S-transferase placental form positive (GST-P$^{+}$) foci, apoptosis, DNA fragmentation and apoptosis related proteins were determined respectively. The formation of GST-P$^{+}$foci was significantly decreased by mistletoe extract or lectin treatment. Although there was no effect on apoptosis and DNA fragmentation in hepatic tissue by mistletoe extract or lectin treatment, caspase-9 and fas-L were increased. These results suggest that Korean mistletoe extract and lectin have a potential to inhibit hepatocarcinogenesis by increasing apoptosis.sis.

HISTOMORPHOLOGIC CHANGE AND REGENERATION OF THE ADVENTITIA-STRIPPED AND FROZEN RAT FEMORAL ARTERY:HISTOLOGIC, HISTOMORPHOMETRIC, IMMUNOHISTOCHEMICAL AND SCANNING ELECTRON MICROSCOPIC STUDY (백서 대퇴동맥 외막층 제거후 혈관동결시 조직형태학적 변화 및 재생에 관한 연구)

  • Kim, Tae-Deug;Kim, Kyung-Wook;Lee, Jae-Hoon;Kim, Chang-Jin
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.25 no.4
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    • pp.281-294
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    • 1999
  • Vasospasm causes microvascular surgery to fail as a main factor in the loss of transferred flap dye to the diminution of blood flow in reconstruction surgery. Although there has been extensive research to resolve the vasospasm problem, no one has reached an ideal solution to date. However, cryotherapy, which is often used for destruction of tumor lesions, is being presented as a new way of releasing vasospasm. After making a histomorphometric measurement on vasodialation during the course of 1, 3 and 7 days, 2 and 4 weeks, and 5 months periods and observing the change of blood vessel in a histologic, immunohistochemical, and scanning electronic microscopic approach, the results were as follows : 1. Vascular inner diameters of the experimental 1 and 3 days groups were measured $476.3{\pm}28.20{\mu}m$, $497.15{\pm}48.79{\mu}m$ respectively showing statistically meaningful vasodilation(P<0.05), which continued by the experiment 4 weeks group. However, in the experimental 5 months group, the vascular inner diameter appeared similar to the control groups. Even though the thickness of smooth muscular layers come out to be thinner in all the experimental groups compared to the control group, it was difficult to find any statistical meaningfulness. In addition, the vascular external diameters of every experimental groups were shown to be longer than the control group. 2. In light microscopic view, severe injury was evident on the smooth muscular layer cell from the experimental 1 day group, started recovering partially from the experimental 7 days group, and was mostly restored in the experimental 4 weeks group and layer of adventitial stripping were nearly recoverd 2 weeks group. 3. The PCNA positive cells of smooth muscular layer were observed from the experimental 7 days group and had a tendency to increase by the experimental 2 weeks group. In the experimental 4 weeks and 5 months group, the number of PCNA possitive cells observed was comparable to the control group. 4. ${\alpha}$-SMA level of smooth muscular layer cells, having been significantly lower than the control group in the severly damaged experimental 1 day group. It was seen to be increased in the experimental 7 days group and turned out to show similar ${\alpha}$-SMA level in 4 weeks to the control group. 5. In the view of SEM, the endothelial cells were destructed and falling off, and also present the appearance of flattening in the experiment 1 day group. The endothelial layer cells started partially recovering from the 7 days group after the freezing injury. On 4 weeks and 5 months, the endothelial cells were fully coverd the damaged area, also it's appearance is similar to control group. In conclusion, the vascular freezing after the removal of adventitia caused damages to smooth muscular layer cells, and brought about vasodilation, which continued by the 4th week. The smooth muscular layer cells started partially reviving from the 7rd day after the damage by vascular freezing, and recovered their similar figure to the control group's 4 weeks later. This was considered the result of cells which surround the damaged blood vessel being influxed into the smooth muscular layers. Therefore, this local freezing injury on the blood vessel was thought to be applied clinically to relieve severe vasospasm which cannot be treated by vasodilation drug, a microvascular surgery.

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The effect of silk fibroin and rhBMP-2 on bone regeneration in rat calvarial defect model (백서 두개골결손모델에서 실크단백과 골형성단백 이식체가 골재생에 미치는 영향)

  • Nam, Jeong-Hun;Noh, Kyung-Lok;Pang, Eun-O;Yu, Woo-Geun;Kang, Eung-Sun;Kweon, Hae-Yong;Kim, Seong-Gon;Park, Young-Ju
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.36 no.5
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    • pp.366-374
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    • 2010
  • Introduction: This study evaluated the capability of silk fibroin (SF) and recombinant human bone morphogenetic protein-2 loaded SF (SF-BMP) as a bone defect replacement matrix when grafted in a calvarial bone defect of rats in vivo. Materials and Methods: A total 70 calvarial critical size defects (5.0 mm in diameter) made on 35 adult female Sprague-Dawley rats were used in this study. The defects were transplanted with (1) rhBMP-2 loaded silk fibroin graft (SF-BMP: 0.8+$10\;{\mu}g$), (2) Silk fibroin (SF: $10\;{\mu}g$), and (3) no graft material (Raw). The samples were evaluated with soft x-rays, alkaline phosphatase activity, calcium/phosphate quantification, histological and histomorphometric analysis at postoperative 4 and 8 weeks. Results: The SF-BMP group ($48.86{\pm}14.92%$) had a significantly higher mean percentage bone area than the SF group ($24.96{\pm}11.01%$) at postoperative 4 weeks.(P<0.05) In addition, the SF-BMP group ($40.01{\pm}12.43%$) had a higher % bone area at postoperative 8 weeks than the SF group ($33.26{\pm}5.15%$). The mean ratio of gray scale levels to the host bone showed that the SF-BMP group ($0.67{\pm}0.08$) had a higher mean ratio level than the SF group ($0.61{\pm}0.09$) at postoperative 8 weeks. These differences were not statistically significant.(P=0.168 and P=0.243, respectively) The ratio of the calcium and phosphate contents of the SF-BMP ($0.93{\pm}0.22$) group was lower than that of the SF ($1.90{\pm}1.42$) group at postoperative 4 weeks. However, the SF-BMP group ($0.75{\pm}0.31$) had a higher Ca/$PO_4$ ratio than the SF ($0.68{\pm}0.04$) at postoperative 8 weeks. These differences were not statistically significant.(P=0.126 and P=0.627, respectively) For the bone-specific alkaline phosphatase (ALP) activity, which is recognized as a reliable indicator of the osteoblast function, the SF-BMP ($23.71{\pm}8.60\;U/L$) groups had a significantly higher value than the SF group ($12.65{\pm}6.47\;U/L$) at postoperative 4 weeks.(P<0.05) At postoperative 8 weeks, the SF-BMP ($21.65{\pm}10.02\;U/L$) group had a lower bone-specific ALP activity than the SF group ($16.72{\pm}7.35\;U/L$). This difference was not statistically significant.(P=0.263) For the histological evaluation, the SF-BMP group revealed less inflammation, lower foreign body reactions and higher bone healing than the SF group at postoperative 4 and 8 weeks. The SF group revealed more foreign body reactions at postoperative 4 weeks. However, this immunogenic reaction decreased and the remnant of grafted material was observed at postoperative 8 weeks. For histomorphometric analysis, the SF-BMP group had a significantly longer bone length to total length ratio than those of the SF group at postoperative 4 and 8 weeks.(P<0.05) Conclusion: The rhBMP-2 loaded silk fibroin graft revealed fewer immunoreactions and inflammation as well as more new bone formation than the pure silk fibroin graft. Therefore, silk fibroin may be a candidate scaffold for tissue engineered bone regeneration.