In Vitro에서 골 형성과 흡수에 대한 Endothelin-1의 영향

사석진,안도환 고신대학교 의과대학 2010 고신대학교 의과대학 학술지 Vol.25 No.2

Background: Balanced regulation of the receptor activator of nuclear factor-kB (RANK) ligand (RANKL) and osteoprotegerin (OPG) by osteoblasts is important for osteoclastogenesis. Sex hormones, glucocorticoids, and prostaglandin E2 (PGE2) are known to modulate osteoblast proliferation and osteoclast formation. Endothelin (ET)-1 is a mitogen as well as a strong vasoconstrictor.It stimulates proliferation of osteoblasts, but its effects on differentiation are controversial. In addition, little is known about ET-1 regulation of osteoclast formation. Thus, the present study was undertaken to investigate whether ET-1 canregulate the expression of RANKL and OPG genes in osteoblasts and affect RANKL-induced osteoclastogenesis. Methods: Osteoblasts were derived from neonatal calvariae and monocytic preosteoclasts from the bone marrow of adult mice, respectively. Cells were cultured in a-minimum essential medium containing 10 nM ET-1. The gene expressions of RANKL and OPG in osteoblasts and RANK in preosteoclasts were measured by real-time RT-PCR. Mineralization by osteoblasts was determined by Alizarin-red staining. Osteoclastogenesis was examined using tartrate-resistant acid phosphatase (TRAP) staining and resorption pit assay. Results: Osteoblasts expressed both ETA and ETB receptors (ETAR and ETBR). ET-1 (10 nM) increased osteoblast proliferation 1.6-fold compared with the control after 3 days in culture and stimulated differentiation, which was indicated by increased formation of mineralized matrix. Proliferation and differentiation of osteoblasts was blocked by 1 uM BQ123, an ETAR antagonist. ET-1 suppressed RANKL gene expression by 50% but did not affect OPG gene expression, and thus reduced the ratio of RANKL to OPG mRNA. PGE2 production by osteoblasts was increased by ET-1. In preosteoclast cultures, ET-1 had little effect on RANK mRNA expression and suppressed RANKL-induced osteoclastogenesis by decreasing the number of TRAP-positive osteoclasts and the resorbed areas. Conclusions: These findings demonstrate that ET-1 can increase bone formation by stimulating proliferation and differentiation of osteoblasts and decrease bone resorption by suppressing RANKL-induced osteoclastogenesis as well as suppressing RANKL mRNA expression In vitro에서 골 형성과 흡수에 대한 endothelin-1의 영향 배경: 조골세포에서 생성되는 receptor activator of nuclear factor-kB ligand (RANKL)과 osteoprotegerin (OPG) 사이의 균형은 파골세포 형성에 매우 중요하다. Enodothelin-1(ET-1)은 강력한 혈관수축제일뿐만 아니라 다양한 세포에서 세포 증식을 촉진시키는 물질이다. 그러나 ET-1의 조골세포의 분화에 대한 영향은 상반될 뿐만 아니라 ET-1이 조골세포의 RANKL과 OPG의 생성을 변화시켜 간접적으로 파골세포 형성에 영향을 주는지 혹은 파골세포의 분화에 직접적으로 영향을 주는지에 대해서는 연구된 바 없다. 따라서 본 연구에서는 ET-1에 의한 조골세포의 분화와 RANKL 및 OPG 유전자 발현이 파골세포 형성에 미치는 영향을 조사하였다. 재료와 방법: 조골세포는 생쥐의 두개골로부터 그리고 파골전구세포는 골수로부터 각각 분리하였다. 이들 세포들을 10 nM ET-1이 포함된 α-MEM 배지에서 배양하였다. 유전자의 발현은 real-time RT-PCR을 통해 정량적으로 측정하였다. 조골세포의 분화는 alizarin-red 염색으로, 파골세포의 형성과 활성은 tartrate-resistant acid phosphatase 염색과 골 흡수와(resorption pit)의 크기로 평가하였다. 결과: ET-1은 조골세포의 증식과 분화를 촉진시켰으며, 이는 ETA 수용체를 통해 이루어졌다. 조골세포에서 ET-1은 PGE2 생성을 크게 증가시켰을 뿐만 아니라 RANKL 유전자 발현을 대조군에 비해 50% 감소시켰다. 그러나 OPG 유전자의 발현에 영향을 주지 않았다. 파골 전구세포에서 ET-1은 RANK 유전자의 발현에 영향을 주지 않았지만 RANKL에 의해 유도된 파골세포 형성과 활성은 억제시켰다. 결론: ET-1은 조골세포의 증식과 분화를 촉진시키고 RANKL mRNA 발현을 감소시켜 파골세포 형성을 간접적으로 억제한다. 또한 RANKL에 의해 유도되는 파골세포 형성과 활성을 직접적으로 억제한다. 이러한 결과로 미루어 ET-1은 골형성은 촉진하고 골흡수는 억제할 것으로 사료된다.

고립성 골낭종의 치료

김재도,사석진,이덕희 고신대학교(의대) 고신대학교 의과대학 학술지 1997 고신대학교 의과대학 학술지 Vol.12 No.1

-Abstract- Simple bone cyst is a true benign lesion of primary intraosseous origin that almost always begins on the metaphyseal side of the epiphyseal plate of long bone in children. The pathogenesis is yet undetermined, but its clinical importance is stressed due to prone to pathologic fracture and high recurrence rate. Regardless the variable methods of treatment have been performed, but there is no an established mode of treatment. Therefore we evaluated the results according to the mode of treatment. We have studied 9 cases of simple bone cyst which where treated from Jun. 1987 to May 1995. The average duration of follow-up was 2 years. 3 cases were treated with curettage and bone graft, and 6 cases were treated with steroid injection. The clinical results were as follows: 1. Age distribution was from 4 years to 13 years (mean age 8.4 years). There were 7 boys and 2 girls. The male to female ratio was 3.5 to 1. 2. Skeletal location was proximal humerus in 5 cases, proximal femur & diatal femur, proximal radius and proximal ulna in 1 case respectively. 3. In cases of curettage and bone graft, 2 cases were healed and 1 case was recurred. Then, additional steroid injection was performed. And in 6 cases of steroid injection, 3 cases were healed, 2 cases were improved and 1 case was recurred. However, all cases were completely healed. 4. There was no difference in the results between treatment of steroid injection and curettage & bone graft. And age, sex, location and type of the cyst did not seem to influence the results of treatment. In conclusion, the steroid injection is a simple and safe procedure, and more favorable results can be expected than other surgical methods. The steroid injection should be repeated in persisting or recurrent cases, but it is highly recommended as the method of choice in the initial treatment of simple bone cyst without accompanying a pathologic fracture.

고립성 골낭종의 치료

김재도,사석진,이덕희 고신대학교 의학부 1997 高神大學校 醫學部 論文集 Vol.12 No.1-2

Simple bone cyst is a true benign lesion of primary intraosseous origin that almost always begins on the metaphyseal side of the epiphyseal plate of long bone in children. The pathogenesis is yet undetermined, but its clinical importance is stressed due to prone to pathologic fracture and high recurrence rate. Regardless the variable methods of treatment have been performed, but there is no an established mode of treatment. Therefore we evaluated the results according to the mode of treatment. We have studied 9 cases of simple bone cyst which where treated from Jun. 1987 to May 1995. The average duration of follow-up was 2 years. 3 cases were treated with curettage and bone graft, and 6 cases were treated with steroid injection. The clinical results were as follows : 1. Age distribution was from 4 years to 13 years (mean age 8.4 years). There were 7 boys and 2 girls. The male to female ratio was 3.5 to 1. 2. Skeletal location was proximal humerus in 5 cases, proximal femur & diatal femur, proximal radius, and proximal ulna in 1 case respectively. 3. In cases of curettage and bone graft, 2 cases were healed, and 1 case was recurred. Then, additional steroid injection was performed. And in 6 cases of steroid injection, 3 cases were healed, 2 cases were improved, and 1 case was recurred. However, all cases were completely healed. 4. There was no difference in the results between treatment of steroid injection and curettage & bone graft. And age, sex, location, and type of the cyst did not seem to influence the results of treatment. In conclusion, the steroid injection is a simple and safe procedure, and more favorable results can be expected than other surgical methods. The steroid injection should be repeated in persisting or recurrent cases, but it is highly recommended as the method of choice in the initial treatment of simple bone cyst without accompanying a pathologic fracture.

고립성 골낭종의 치료

김재도,사석진,이덕희 高神大學 醫學部 1997 高神大學醫學部 論文集 Vol.12 No.1-2

가토의 대퇴골에서 이식한 자가골의 체외 방사선 조사에 따른 조직 형태학적 변화

김재도,임언,사석진 고신대학교(의대) 고신대학교 의과대학 학술지 1999 고신대학교 의과대학 학술지 Vol.14 No.1

Background : Skeletal reconstruction is an inevitable procedure when limb sparing operation is indicated in the treatment of the malignant bone tumor of extremities. After a bone tumor is widely resected, several procedures have been used such as tumor prosthesis, allograft, allograft-prosthesis composite, ordinary autograft, recycling autograft (autoclave, low heat treated, extracorporeal irradiated) to fill the large osseous defect. Methods : The rabbits were divided into three groups according to the irradiation dose given to the separated femoral segment : 0 (control), 20, and 50 Gy, respectively. A cortical bone segment of the unilateral femoral diaphysis in rabbit was resected, irradiated extracorporeally, and reimplanted immediately as an autograft, and then examined over a 8 week period by plain roentgenography, bone scan, gross and histologic findings. Results : There were no definite difference among three groups. : The osteotomy site was grossly stable with external callus formation.On plain roentgenography, the gap of osteotomy site was disappeared with abundant external callus formation. On bone scan, the uptake of radioactive isotope was increased in incorporated site compared with contralateral one. Histologically, the resorption of the graft began on the outer surface of cortex. New bone formation was confined around the osteotomy sites. It was much more prominent on the periosteal side than in the endosteal side in both the control and irradiated groups. Conclusion : These results suggest that bone union is not affected by irradiation of 20 Gy to 50 Gy as used in this study. Therefore, extracorporeal irradiated autogenous bone graft can be used as a limb preserving procedure for the malignant bone tumor.

가토의 대퇴골에서 이식한 자가골의 체외 방사선 조사에 따른 조직 형태학적 변화

김재도,임언,사석진 고신대학교 의학부 1999 高神大學校 醫學部 論文集 Vol.14 No.1-2

Background Skeletal reconstruction is an inevitable procedure when limb sparing operation is indicated in the treatment of the malignant bone tumor of extremities. After a bone tumor is widely resected, several procedures have been used such as tumor prosthesis, allograft, allograft-prosthesis composite, ordinary autograft, recycling autograft(autoclave, low heat treated, extracorporeal irradiated) to fill the large osseous defect. Methods The rabbits were divided into three groups according to the irradiation dose given to the separated femoral segment ; 0 (control), 20, and 50 Gy, respectively. A cortical bone segment of the unilateral femoral diaphysis in rabbit was resected, irradiated extracorporeally, and reimplanted immediately as an autograft, and then examined over a 8-week period by plain roentgenography, bone scan, gross and histologic findings. Results There were no definite difference among three groups. ; The osteotomy site was grossly stable with external callus formation. On plain roentgenography, the gap of osteotomy site was disappeared with abundant external callus formation. On bone scan, the uptake of radioactive isotope was increased in incorporated site compared with contralateral one. Histologically, the resorption of the graft began on the outer surface of cortex. New bone formation was confined around the osteotomy sites. It was much more prominent on the periosteal side than in the endosteal side in both the control and irradiated groups. Conclusion These results suggest that bone union is not affected by irradiation of 20 Gy to 50 Gy as used in this study. Therefore, extracorporeal irradiated autogenous bone graft can be used as a limb preserving procedure for the malignant bone tumor.

선택적 척추신경근 차단술을 통한 방사통의 치료

홍영기(Young Gi Hong),사석진(Sok Jin Sa),김재도(Jae Do Kim) 대한정형외과학회 1997 대한정형외과학회지 Vol.32 No.4