신연 골형성술시 신연속도에 따른 골형성 관여 인자의 발현

지유진(Yu-Jin Jee),김여갑(Yeo-Gab Kim) 대한구강악안면외과학회 2008 대한구강악안면외과학회지 Vol.34 No.3

Distraction osteogenesis is a well-established clinical treatment for limb length discrepancy and skeletal deformities. Appropriate mechanical tension-stress is believed not to break the callus but rather to stimulate osteogenesis. In contrast to fracture healing, the mode of bone formation in distraction osteogenesis is primarily intramembranous ossification. Although the biomechanical, histological, and ultrastructural changes associated with distraction osteogenesis have been widely described, the basic biology of the process is still not well known. Moreover, the molecular mechanisms in distraction osteogenesis remain largely unclear. Recent studies have implicated the growth factor cascade is likely to play an important role in distraction. And current reserch suggested that mechanical tension-stress modulates cell shape and phenotype, and stimulates the expression of the mRNA for bone matrix proteins. The purpose of this study is to examine the pattern of expression of growth factors(TGF-β1, IGF-I, bFGF) and extracellular matrix proteins(osteoclacin, osteonectin) related to osteogenesis by osteodistraction of the mandible in rabbits. 24 rabbits is used for this experiment. Experimental group are gradual distraction(0.7mm, twice/day), acute distraction(1.4mm, twice/day) and control group is only osteotomized. After 5 days latency, osteotomic site is distracted for each 7 days and 3.5 days. Consolidation period is 28 days. The animal is sacrificed at the 3th, 7th, 14th, 28th. The distracted bone is examined by immunohistochemical analysis and RT-PCR analysis. The results obtained from this study were as follow : No significant difference was found on clinical examination according to distraction rate, but gradual distraction was shown to improve regenerate bone formation on radiographic and histologic examination. Growth factors and extracelluar matrix proteins expression increased in distraction group than control group. From these results, it could be stated that graudal distraction is shown to improve and accelerate bone formation and mechanical stress like distraction has considerable effects on osteogenesis related factors. And rabbit is the most appropriate animal model for further reseach on the molecular mechanisms that mediate osteodistraction. It is believed that understanding the biomolecular mechanisms that mediate distraction osteogenesis may guide the development of targeted strategies designed to improve distraction osteogenesis and accelerate bone healing.

백서 하악골에서 신연골형성술시 압축력의 효능에 관한 실험적 연구

강항립(Hang-Rip Kang),김철훈(Cheol-Hun Kim),신상훈(Sang-Hoon Shin),정인교(In-Kyo Chung),김욱규(Uk-Kyu Kim) 대한구강악안면외과학회 2004 대한구강악안면외과학회지 Vol.30 No.5

Distraction osteogenesis is a biologic process of new bone formation between the surfaces of bone segments that are gradually separated by incremental traction. Distraction osteogenesis is clinically applied as a new treatment modality of mandibular hypoplasia or bony defect area in maxillofacial area by many studies of distraction devices and method. But disadvantage of distraction osteogenesis shows unfavorably long consolidation period and relapse tendency. Therefore. this experiment was designed to investigate the effectiveness of combined application of distraction and compression force for improving of bone quality and shortening of treatment period during distraction osteogenesis. Twenty-one Sprague-Dawley rats with 300~350gm were used. These were divided into two group as distraction group and combination group was applied with compression force in the consolidation period. The rat were sacrificed for gross finding, radiographic and histologic findings. at 2, 4 weeks after distraction. The result were follow : 1. On radiographic finding, all experimental groups appeared more radiopacity than control groups both at 2, 4 weeks after distraction. 2. On histologic finding, trabeculae of bone and mature lamellar bone were showed increasingly in experimental group. Ossification occured rapidly. From this study, we may suggest that compression force application in consolidation period during distraction osteogenesis can be useful method improve bone quality and to shorten the treatment period. But more experimental and clinical studis are necessitated on effects of compression force application during distraction osteogenesis.

하악골 신장술 후 신생 골막조직에서의 혈관내피세포성장인자 및 혈관내피세포성장인자 수용체 발현에 대한 연구

황등욱,변준호,박봉욱,김종렬,Hwang, Deung-Uc,Byun, June-Ho,Park, Bong-Wook,Kim, Jong-Ryoul 대한악안면성형재건외과학회 2006 Maxillofacial Plastic Reconstructive Surgery Vol.28 No.6

During distraction osteogenesis, the angiogenic activity is crucial factor in the new bone formation. The aim of this study was to detect the autocrine growth activity in the cellular components of the distracted periosteum with observation of the expression of vascular endothelial growth factor (VEGF) and its receptors following the mandibular distraction osteogenesis. Unilateral mandibular distraction (0.5 mm twice per day for 10 days) was performed in six mongrel dogs. Two animals were sacrificed at 7, 14, and 28 days after completion of distraction, respectively. The distracted lingual periosteum was harvested and processed for immunohistochemical examinations. After then, we observed the expression of VEGF, Flt-1 (VEGFR-1), and Flk-1 (VEGFR-2) in the osteoblasts and immature mesenchymal cells of the distracted periosteum. At 7 days after distraction, the expression of VEGF and its receptors were significantly increased in the cellular components of the distracted periosteum. Up to 14 days following distraction, the increased expressions were maintained in the osteoblastic cells. At 28 days after distraction, the expression of VEGF and its receptors decreased, but VEGF was still expressed weak or moderate in the osteoblastic cells of distracted periosteum. The expression pattern of VEGF and its receptors shown here suggested that VEGF play an important role in the osteogenesis, and these osteoblastic cell-derived VEGF might act as autocrine growth factor during distraction osteogenesis. In the other word, the cellular components in the distracted periosteum, such as osteoblasts and immature mesenchymal cells, might have autocrine growth activity during distraction osteogenesis.

수축력과 신장력을 병용한 골신장술에서의 다양한 힘의 비와 부여시기에 따른 신연골 반응

김욱규,신상훈,정인교,김철훈,허준,윤일,Kim, Uk-Kyu,Shin, Sang-Hun,Chung, In-Kyo,Kim, Cheol-Hun,Huo, Jun,Yun, Il 대한악안면성형재건외과학회 2005 Maxillofacial Plastic Reconstructive Surgery Vol.27 No.5

The purpose of this study was to identify the effectiveness of the modified distraction osteogenesis (DO) method with the concept of overdistraction and compression stimulation which have been previously suggested by the authors in 2002 and to explore the optimal distraction-compression ratio and appropriate latency period for the compression force application during consolidation. The experimental specimens were assessed with radiography, histologic findings, and dual energy x-ray absorptiometry (DEXA) after the conventional DO method and the modified DO technique had been applied on rat mandibles. Total 60 rats were used for the study. In experimental group of 54 adult rats, mandibular osteotomies between the first and second molar areas were performed and customized external distractors were applied. The surgeries on 6 rats of control group also were done with same osteotomy technique and DO device application. Final amount of distraction was set-up as 2 mm on both groups. But, in a experimental group of 54 rats, distraction osteogenesis with a compression force were performed with the different distraction-compression ratio and variant latency periods for compression. The three ratio-subgroups were made as distraction 4 mm group with compression 2 mm, distraction 3 mm group with compression 1 mm, and distraction 2.5 mm group with compression 0.5 mm. In addition, The three subgroups with 3, 7, 11 days latency period prior compression were allocated on each ratio-subgroups. Total 9 subgroups consisted of 6 rats on each subgroup. In control group of 6 rats, conventional distraction technique were routinely performed. The rats of control groups were sacrificed on postoperative 3, 6 weeks after 2 mm distraction. The rats in the experimental groups also were sacrificed on the same euthanasia days of control groups to compare the wound healing. Final available specimens were 55 rats except 5 due to osteomyelitis, device dislodgement. Distraction-compression combined group on 6 weeks generally had showed increased bone mineral density than the same period group of conventional distraction technique on the DEXA study. More matured lamellar bone state and extended trabecular pattern in the combined group than those of control group were also observed in the histologic findings on 6 weeks. In the distraction-compression combined groups, the bone density of 2.5 mm distraction subgroups with 0.5 mm compression showed the highest value on the DEXA study among various force ratio groups. In the distraction-compression combined groups, the bone density of 3 day latency period subgroups showed the highest value on the DEXA study among various latency period groups for the compression application. From this study, we could deduce that 1/5 force ratio for the compression versus distraction, 3 day latency period prior compression application would be the most effective condition if modified distraction osteogenesis technique might be applicable. The modified DO method with a compression force may improve the quality of bone regenerate and shorten total treatment period in comparison with conventional DO technique clinically.

성견 하악골 신장술 후 신생골조직에서 Osteopontin 발현에 대한 연구

변준호(June-Ho Byun),박봉욱(Bong-Wook Park),성일용(Iel-Yong Sung),조영철(Yeong-Cheol Cho),김종렬(Jong-Ryoul Kim) 대한구강악안면외과학회 2006 대한구강악안면외과학회지 Vol.32 No.5

Purpose: This study was aimed at evaluating the histological changes of new bone and expression of osteopontin (OPN) after mandibular distraction osteogenesis. Materials and Methods: Unilateral mandibular distraction (0.5 mm twice per day for 10 days) was performed in eight adult dogs. Two animals were sacrificed at 7, 14, 28 and 56 days after completion of distraction, respectively. The distracted bones and contralateral non-distracted control bones were harvested and processed for histological and immunohistochemical examinations. Results: The new bone was arranged to tension direction after distraction osteogenesis. 7 days after distraction, numerous osteoblasts lining the immature trabecular bone and fibroblast-like cells in the fibrous intrezone were observed. 14 days after distraction, the new formed trabecular bones were thickened compared with 7 days after distraction. 28 days after distraction, the fibrous interzone was almost filled with newly calcified bone, and it was more hardened at 56 days after distraction. Increased OPN signals detected in the osteoblasts lining the trabecular bone and fibroblast-like cells in the fibrous interzone at 7 and 14 days after distraction. At 28 days after distraction, the OPN was weakly expressed in the osteoblasts, and it was not detected in all cellular components of distracted bone at 56 days later of distraction. Conclusions: After distraction osteogenesis, the distracted zone was completely calcified during the 56 days of consolidation period. In this study, the staining intensity of OPN increased in the osteoblasts and fibroblast-like cells at 7 and 14 days after distraction. The expression pattern of this protein shown here suggested that OPN play an important role in the osteogenesis during the early consolidation period.

가토 하악골에서 신연 골형성술시 신연속도에 따른 TGF-$\beta$1, IGF-I, bFGF의 발현

신선아,지유진,송현철,Shin, Sun-A,Jee, Yu-Jin,Song, Hyun-Chul 대한악안면성형재건외과학회 2005 Maxillofacial Plastic Reconstructive Surgery Vol.27 No.3

Distraction osteogenesis is a technique of lengthening bone including soft tissue by gradual separation of surgically divided bone surfaces. Although the biomechanical, histological, and ultrastructural changes associated with distraction osteogenesis have been widely described, the molecular mechanisms governing the formation of new bone in distracted bone segments remain largely unclear. However, such information has significant clinical implications because it may enable targeted therapeutic manipulations designed to accelerate osseous regeneration. The purpose of this study was to evaluate the expression of TGF-$\beta$1, IGF-I and bFGF in distraction osteogenesis according to different distraction rates in a rabbit's mandible. When twenty-four adult rabbits underwent open osteotomy between the premolar and mental foramen, an external bilateral distraction device was applied. Latency was allowed for five days before distraction. Three different distraction rates were 0.7 mm/day (A, n=8), 1.4 mm/day (B, n=8) and 2.4 mm/day (C, n=8). The distraction device was activated with the same distraction rhythms of twice a day until 4.9 mm (A & B group) and 8.4 mm (C group) length gains was achieved. The animals were sacrificed at postoperative 3, 7, 14 and 28 days. The bony specimens were stained with H&E for histologic examination, and RT-PCR analysis was done for the identification of the expression of TGF-$\beta$1, IGF-I and bFGF. The results obtained from this study were as follows : The 0.7 mm/day and 1.4 mm/day distraction rate groups were shown to improve regenerative bone formation on radiographic and histologic examination. Also, TGF-$\beta$1, IGF-I and bFGF expression increased in the 0.7 mm/day and 1.4 mm/day distraction rate groups. But the 2.4 mm/day distraction rate group specimen was different with adjacent normal bone and hardly expressed of growth factors. These findings suggest that improved new bone formation in the 0.7 mm/day and 1.4 mm/day distraction rates is associated with enhanced expression of TGF-$\beta$1, IGF-I and bFGF by mechanical tension stress. Additionally, the 0.7 mm/day and 1.4 mm/day distraction rate groups were significantly different from the 2.4 mm/day distraction rate group in the expression of growth factors. According to the above results, it seems possible to apply a distraction rate of up to 1.4 mm/day a day in rabbit's mandible. And further studies are needed to evaluate growth factors of TGF-$\beta$1 and IGF-I, which are excellent in expression.

성견에서 하악골 신장술 후 하악과두 연골의 조직학적 변화와 Matrix Metalloproteinase-2 (MMP-2)와 Tissue Inhibitor of Matrix Metalloproteinase-2 (TIMP-2)의 발현

변준호,박봉욱,조영철,성일용,손재희,김종렬,Byun, June-Ho,Park, Bong-Wook,Cho, Yeong-Cheol,Sung, Iel-Yong,Son, Jae-Hee,Kim, Jong-Ryoul 대한악안면성형재건외과학회 2006 Maxillofacial Plastic Reconstructive Surgery Vol.28 No.5

Purpose : This study was to clarify the changes in mandibular condyle after unilateral mandibular distraction osteogenesis throughout histological changes and expression of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of matrix metalloproteinase-2 (TIMP-2). Materials & Methods : Intraoral distractors were placed via submandibular incision in 8 dogs. Two unoperated animals served as controls. Distraction was performed five days after osteotomy as a rate of 0.5 mm twice per day for 10 days. Two animals were sacrificed on 7, 14, 28, and 56 days after completion of distraction, respectively. Ipsilateral condyles were harvested and processed for histological and immunohistochemical examinations. Results : The condyle cartilage is separated into four layers: fibrous layer, proliferative layer, hypertrophic layer, and calcified layer. At 7 days and 14 days after distraction, the condylar cartilage showed the decreased thickness of the articular cartilage and reduced cellularity. At 28 days after distraction, there was an increase in cellularity of fibrous, proliferative, and hypertrophic layer. However, it demonstrated reduced cellularity compared to the control. At 56 days of after distraction, the articular cartilage was an almost normal histologic structure. Positive Safranin-O staining, indicative of sulfated proteoglycans, was examined in the condylar cartilge of nonloaded control. At 7 days and 14 days after distraction, the sulfated proteoglycans is almost completely depleted from the noncalcified part of the condylar cartilage. At 28 days after distraction, there was an increase in Safranin-O staining intensity. However, the staining intensity of the experimental condyle was weaker than that of the control. At 56 days of after distraction, the condylar cartilage showed almost normal Safranin-O staining pattern. In control condyle, MMP-2 immunostaining was seen in fibrous, proliferative, and hypertrophic layer of condylar cartilage, however, it demonstrated lack of staining in fibrous and proliferative layer. At 7 days and 14 days after distraction, strong MMP-2 immunoreactivity was seen in the fibrous, proliferative and hypertrophic layer of the condylar cartilage. At 28 days after distraction, MMP-2 immunostaining was seen in the fibrous and hypertrophic layer of condylar cartilage, however, their immunoactivity was reduced. At 56 days after distraction, MMP-2 immunoreactivity showed almost normal immunostaining pattern. In control condyle, TIMP-2 immunostaining was primarily seen in fibrous and hypertrophic layer of condylar cartilage, however, it demonstrated lack of staining in proliferative layer. At 7 days after distraction, very weak TIMP-2 immunoreactivity appeared in fibrous, proliferative and hypertrophic layer of the condylar cartilage. At 14 days after distraction, weak TIMP-2 immunoreactivity was seen in the fibrous, proliferative and hypertrophic layer of the condylar cartilage. At 28 days after distraction, TIMP-2 immunoreactivity was increased in the fibrous and hypertrophic layer of condylar cartilage. At 56 days after completion of distraction, TIMP-2 immunoreactivity showed almost normal immunostaining pattern. Conclusions : The results show that short-term outcome of physiologic distraction osteogenesis may lead to degenerative changes in the condylar cartilage. These alterations in the condylar cartilage may be considered as a pressure-related degeneration of the cartilage tissue. However, the long-term results suggest that the condylar cartilage display repair activity after mandibular distraction osteogenesis.

가토 하악골 골신장 후 맥동전자기장이 조기 골경화에 미치는 효과에 대한 연구

황경균,조태형,송윤미,김도균,한성희,김인숙,황순정,Hwang, Kyung-Kyun,Cho, Tae-Hyung,Song, Yun-Mi,Kim, Do-Kyun,Han, Sung-Hee,Kim, In-Sook,Hwang, Soon-Jung 대한악안면성형재건외과학회 2007 Maxillofacial Plastic Reconstructive Surgery Vol.29 No.2

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.

구순구개열 환자에서의 상악골 열성장의 골신장술을 위한Internal distraction device와 Rigid external distraction의 비교

팽준영,명훈,황순정,서병무,최진영,이종호,정필훈,백승학,김명진 대한악안면성형재건외과학회 2005 Maxillofacial Plastic Reconstructive Surgery Vol.27 No.4

Distraction osteogenesis for the advancement of hypoplastic maxilla of cleft patients has shown successful results. In this report, rigid external distraction(RED) system and internal distraction device were used for maxillary advancement. Each system has its advantages and disadvantages. Larger amount of advancement can be achieved with RED system. But complex external device may give patients psychological stress. Internal device is invisible. However its distraction amount have limitation for the advancement (< 20mm) and the vector cannot be changed freely during distraction. The authors treated five cleft patients with maxillary hypoplasia(three with RED system and two with internal distractor). Their results were satisfactory. We present the pros and cons of RED and internal system for maxillary distraction osteogenesis.

개의 하악에서 막성골 중첩이식후 신연골 형성에 관한 연구

박세정,이동훈,이정형,조병채,백봉수 大韓成形外科學會 1999 Archives of Plastic Surgery Vol.26 No.3

The purpose of this study was to investigate the possibility of distraction osteogenesis in membranous onlay bone graft on the mandible and to clarify the histology of bone repair during distraction osteogenesis in the membranous onlay bone in a dog model. Four dogs, 5 months of age at the beginning of the experiment, were used for this study. The zygomatic arch was exposed in the subperiosteal plane and the full-thickness zygomatic arch was harvested to 3 centimeters in length. The lateral surface of the mandibular body was exposed in the subperiosteal plane and the membranous onlay bone graft was performed with firm contact using screws. The osteotomy on the membranous onlay bone graft and underlying mandibular body was carried down week 1 in dog 1, week 2 in dog 2, week 3 in dog 3, and week 4 in dog 4 after membranous onlay bone graft. The external distraction device was applied to the mandibular body. Mandibular distraction was started 7 days after the operation at a rate of 1mm per day for a total of 10 mm distraction over 10 days. After completion of distraction, the distraction device was left in place for 6 weeks bony consolidation of the distracted area. Radiographs were carried out at 2 weeks, 4 weeks and 6 weeks after distraction. New bone between the native underlying mandibular segments was generated in the distracted zone in all dogs. The new bone between the native underlying mandibular segments was generated in the distracted zone in all dogs. The new bone between segments of membranous onlay bone graft was not generated in dog 1, but it was generated in dogs 2, 3 and 4. However, in dog 2 and 3, the new bone between segments of the distracted membranous onlay bone graft presented less firmness with fibrous tissue than that of the native underlying mandibular segment. Histologically, the distracted gap between segments of the membranous onlay bone graft was composed of much fibrous tissue in the central zone while activated osteoblastic cells formed new bone in the margins of the distracted gap in dogs 2 and 3. In dog 4, there were abundant osteoblastic activities in the distracted gap and the new bone appeared as nearly-normal cortical bone. In conclusion, these findings suggested that membranous onlay bone graft had an osteogenic capacity and that distraction osteogenesis was possible in membranous onlay bone graft.