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Differential Expression of STATs and Its Function in Osteogenesis of Mesenchymal Stem Cells.

Ji Suk HWANG ; Kyung Mi LEE ; Ho Sun JUNG ; Jin Woo LEE ; Yun Rak CHOI

Journal of Korean Orthopaedic Research Society.2013;16(1):1-9.

PURPOSE: To investigate the role of STAT5 during osteogenesis differentiation of human mesenchymal stem cells(hMSC). MATERIALS AND METHODS: To assess the expression pattern of STATs during osteogenic differentiation, we performed western blot analysis and RT-PCR. By RNA interference, we have performed effect of STAT5A and STAT5B in osteogenesis. As a result of Luciferase assay, the promoter activity of DLX5 was decreased by STAT5A. RESULTS: To assess the expression pattern of STATs during osteogenic differentiation, we have performed western blot and RT-PCR after 14 day differentiation period. STAT1, 2, 3 and 4 showed no change in expression level during differentiation, while STAT5A and STAT5B displayed steady increase compared to the control. When STAT5A was knock down, the level of osteogenesis was increased. The transcriptional activity of DLX5 was decreased about 70% by STAT5A. CONCLUSION: The expression of STAT5A and STAT5B increased during osteogenesis of hMSC. Also, we shown that STAT5A regulated the transcriptional activity of DLX5. These results indicate that STAT5A acts as a pivotal transcription factor in osteogenesis of hMSC.
Blotting, Western ; Humans ; Luciferases ; Mesenchymal Stromal Cells* ; Osteogenesis* ; RNA Interference ; Transcription Factors

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Effect of Traumatic Brain Injury on Healing of Femur shaft Fracture.

Myung Ho KIM ; Moon Jip YOO ; Hee Gon PARK ; Yeon Jun KIM

Journal of Korean Orthopaedic Research Society.2010;13(2):88-94.

PURPOSE: Among the fracture patients, there is a tendency to form more callus and get fracture united earlier in groups with traumatic brain injury. This retrospective study is to evaluate the factors that might accelerate the bone formation by comparing two groups in serologic tests, clinical and radiologic results. MATERIALS AND METHODS: From March 2001 to July 2009, femur shaft fracture patients were divided in two groups 1) without traumatic brain injury (32cases), 2) fracture with traumatic brain injury combined (30cases). We evaluated the routine serologic exams, amount of callus formations during the follow up period. RESULTS: There was no statistical difference in WBC, CRP, total calcium, LDH level between two groups, except Alkaline phosphatase level. Amount of callus formation on AP radiograph at the last follow up period was 74.9% in study, 42.6% in control group. Lateral radiograph showed 73.2% of callus formation rate in study group and 32.0% in control group. CONCLUSION: Two groups had no significant difference with the routine serologic exam except Alkaline phosphatase. Group with traumatic brain injury had much more amount of callus formation but there was no evidence of traumatic brain injury accelerate the fracture healing.
Alkaline Phosphatase ; Bony Callus ; Brain ; Brain Injuries ; Calcium ; Femur ; Follow-Up Studies ; Fracture Healing ; Humans ; Osteogenesis ; Retrospective Studies ; Serologic Tests

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Glucocorticoid Mechanism of Inhibition of the Inflammatory Cells in Lumbar Intervertebral Disc Cells Stimulated by TNF-alpha Production of Nuclear Factor-kappaB.

In Soo OH ; Sang Eun PARK ; Jong Min SON ; Jun Young CHUNG ; Young Yul KIM

Journal of Korean Orthopaedic Research Society.2010;13(2):80-87.

PURPOSE: To analyze the action mechanism of NF-kappaB, IkappaB-alpha and effect of the Dexamethasone (DEXA) in mediating this inflammation, after stimulating cultured herniated intervertebral disc cells with TNF-alpha. MATERIALS AND METHODS: After cultured human intervertebral disc cells passaged three times, they were divided into four groups: A control group (A), DEXA treatment group (B), TNF-alpha treated group (C), TNF-alpha and DEXA were treated at the same time (D). IL-6 and IL-1beta gene expression were measured with semi-quantitative RT-PCR. Western blot analysis was performed to measure protein expression of IkappaB-alpha in the above groups for 10 minutes, 1 hour, 2 hours. In addition, in order to explain the mechanism of NF-kappaB nuclear binding for each group, the nuclear amount of NF-kappaB binding in the nucleus is measured by EMSA. RESULTS: In RT-PCR, expression of IL-6 and IL-1beta was greatest in group C, followed by group D, group A. IkappaB-alpha expression of the group treated with DEXA was not detected in Western blot results within 10 minutes. However, if stimulated by TNF-alpha, the DEXA was not inhibited of IkappaB-alpha concentration. After 1 hour and 2 hours, IkappaB-alpha levels were expressed by cells autonomously (autoregulatory induction). EMSA results expression levels in nuclear protein was maintained in accordance with protein expression. CONCLUSIONS: Our study shows that DEXA inhibits the production of mediators such as inflammatory IL-6 and IL-1beta, however, may not inhibit the transcription of NF-kappaB stimulated by TNF-alpha.
Blotting, Western ; Dexamethasone ; Gene Expression ; Humans ; I-kappa B Proteins ; Inflammation ; Interleukin-6 ; Intervertebral Disc ; Negotiating ; NF-kappa B ; Nuclear Proteins ; Tumor Necrosis Factor-alpha

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Effect of Curettage and DBM-CaSO4 Graft for the Treatment of Ischemic Necrosis of the Capital Femoral Epiphysis in Immature Pigs.

Ki Seok LEE ; Sun Yong KIM ; In Ho CHOI

Journal of Korean Orthopaedic Research Society.2010;13(2):68-79.

PURPOSE: The purpose of this study was to determine the effect of curettage and DBM complex graft as a new treatment modality for LCP disease using piglet capital femoral epiphysis ischemic necrosis model. MATERIALS AND METHODS: Five to six weeks old piglets were used for the experiment. Ischemic necrosis of the capital femoral epiphysis was surgically induced by cervical ligation on both sides. Three weeks following ischemic insult, the left hip joint was approached medially. About 15% of the necrotic capital femoral epiphysis was curetted through a window which was opened at medial cervical cortex, then, demineralized bone matrix complex was engrafted. The right femoral heads served as controls. Piglets were sacrificed three, six, nine, and twelve weeks following were harvested for histologic examination. RESULTS: In control group, photomicrographs of specimens showed central necrosis and fibrovascular invasion in capital femoral necrosis at three weeks after ischemic insult. Six, nine, and twelve weeks following ischemic insult, fibrovascular invasion advanced without noticeable new bone formation and collapse of femoral head progressed. At twelve weeks, definite coxa plana developed. In curettage and DBM complex graft group, there was evident new bone formation observed in the site of DBM complex graft. At three weeks, new bone formation along with fibrovascular invasion was observed around the engrafted DBM complex mainly in the cervical metaphyseal area. At six and nine weeks, new bone formation progressed into the engrafted DBM complex in the cervical metaphysis and around the engrafted DBM complex in the capital femoral epiphysis. At twelve weeks, new bone along with new cartilage formation was observed in the capital femoral epiphysis. CONCLUSION: In conclusion, curettage and DBM complex graft is thought to be an effective treatment modality that promote regeneration of ischemic necrosis of capital femoral epiphysis.
Bone Matrix ; Cartilage ; Curettage ; Epiphyses ; Head ; Hip Joint ; Legg-Calve-Perthes Disease ; Ligation ; Necrosis ; Osteogenesis ; Regeneration ; Swine ; Transplants

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Evaluation the Reliability of Singh Index in Elderly Patients with Proximal Femoral Fractures Using Digital Radiographic Image.

Ho Hyun YUN ; Jong Woo KANG ; Guen Young LEE ; Jae Wuk LEE ; Ju Won LEE ; Hyoung Won JANG ; Sung Chul PARK

Journal of Korean Orthopaedic Research Society.2010;13(2):60-67.

PURPOSE: To evaluate the reliability of Singh index (SI) values, determined on image software processed digital radiographs in elderly patients with proximal femoral fractures, with respect to its value as a simple and inexpensive method to evaluate osteoporosis in acute trauma situations. MATERIALS AND METHODS: The authors retrospectively reviewed 210 patients (98 femur neck fractures, 112 intertrochanter fractures) treated between March 2005 and March 2009. Preoperative digital radiographs of each patient were assessed by four observers to determine SI values. The reliability of SI was expressed in terms of intraobserver and interobserver agreements in pairs using Fleiss's overall Kappa, Stuart's tau-c index, and Kendall's coefficiency of concordance. RESULTS: Fleiss's overall kappa values for intraobserver agreement ranged from 0.278 to 0.586 (mean, 0.452) and did not reach good reliability overall. Fleiss's overall kappa values for interobserver agreement ranged from 0.120 to 0.241 (mean, 0.180), and thus, did not reach acceptable reliability. CONCLUSION: The result suggest that the image adjustment tools provided by digital radiography cannot improve the usefulness of SI as a simple and inexpensive method of assessing the osteoporosis. Therefore, the reliability of SI good enough to be used for clinical and research work is questionable.
Aged ; Femoral Fractures ; Femoral Neck Fractures ; Hip ; Humans ; Osteoporosis ; Radiographic Image Enhancement ; Retrospective Studies

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Effect of Zoledronic Acid on Early Osseointegration of Porous Implant in Rabbit.

Yoo Joon SUR ; Hyoung Min KIM ; Seok Whan SONG ; Nam Jin YOO

Journal of Korean Orthopaedic Research Society.2010;13(2):53-59.

PURPOSE: The purpose of this study was to determine whether intravenous injection of the zoledronic acid could promote osseointegration of the porous implant inserted into the rabbit medullary cavity. MATERIALS AND METHODS: A rabbit intramedullary osseointegration model was used. A specially designed porous nitinol implant (Bio-Smart, Sungnam, Gyeonggi-do, Korea) was inserted in the right distal femur of twenty six rabbits. They were randomized into the control or the experimental groups. In the control group, an intravenous injection of normal saline 2 ml/kg (Daihan Pharm, Seoul, Korea) was given at the end of the operation. In the experimental group, an intravenous injection of zoledronic acid 0.1 mg/kg (AclastaG(R) 2 ml/kg, Norvatis, Stein, Switzerland) was given at the end of the operation. Six weeks later, all animals were sacrificed and undecalcified histologic sections were prepared. Then, histomorphometric measurement of bone affinity index (%) and bone ingrowth area rate (%) was carried out. RESULTS: Six rabbits were excluded due to death and wound infection. Nine rabbits in the control group and eleven rabbits in the experimental group were included for the analysis. The bone affinity indices were 19.9+/-7.9% in the control group, and 28.4+/-7.2% in the experimental group. Although there was no statistical significance (p=0.056), the bone affinity index of the experimental group was higher than that of the control group. The bone ingrowth area rates were 8.7+/-3.7% in the control group, and 12.1+/-4.1% in the experimental group (p=0.046), indicating zoledronic acid had an positive effect on the promotion of bone ingrowth into the porous implant. CONCLUSION: In our rabbit intramedullary osseointegration model, intravenous injection of the zoledronic acid promoted early osseointegration of the porous implant. Zoledronic acid might be useful to promote the early osseointegration of the joint replacement implants.
Alloys ; Animals ; Diphosphonates ; Femur ; Imidazoles ; Injections, Intravenous ; Joints ; Osseointegration ; Rabbits ; Wound Infection

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Reduction of Broach Rotation for Versys Fibermetal Tapered Stem during the Femoral Canal Shaping.

Youngbae PARK ; Deuk Soo HWANG ; Kyeongbin LIM ; Yong San YOON

Journal of Korean Orthopaedic Research Society.2007;10(2):83-89.

PURPOSE: Inaccurate femoral canal shaping can result in post-operative complications in hip arthroplasty. We addressed the amount of broach rotation during shaping of the femoral canal and compared it with respect to newly designed broaches which were modified to minimize the rotation. MATERIALS AND METHOD: we designed the broaches that had canal guide which restricts the broach motion such that it is always aligned with the femoral axis while the broach machines the metaphyseal bone. Conventional broaches and the modified broach applied to 5 pair of fresh-frozen cadaver femurs and its spatial motion was measured with motion tracker. Rotations in coronal, saggital and frontal plane during the final 10 mm of broach advance were measured. RESULTS: 2.4..of axial rotation was occurred during final 10 mm advance of broach in the conventional method, which was the largest component of the rotation. Rotation of the broach during machining was decreased to 37% (p=0.075) and 25% (p=0.042) in the sagittal plane and coronal plane, respectively, by proposed method. CONCLUSION: The canal guide in the proposed method significantly reduced the rotation of the broach without any extra incision or measurement devices, resulting in increased accuracy in the femoral canal shaping.
Arthroplasty ; Axis, Cervical Vertebra ; Cadaver ; Femur ; Hip

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The Change of Stress Distribution according to Fixation Devices and Osteotomy Methods in High Tibial Osteotomy.

Se Hyun CHO ; Sung Hwan AN ; Hyung Bin PARK ; Soon Taek JEONG ; Sun Chul HWANG

Journal of Korean Orthopaedic Research Society.2007;10(2):76-82.

PURPOSE: To evaluate the necessity of lateral translation and the primary stability of the fixation devices in a closed wedge high tibial osteotomy. MATERIALS AND METHODS: The authors studied four pairs of high tibial osteotomy: Pair I, lateral translation with blade plate fixation; Pair II, lateral translation with staple fixation; Pair III, no translation with blade plate fixation; Pair IV, no translation with staple fixation. Four models of bovine tibia were taken and analyzed for stress distribution at the osteotomy site under axial loading. After axial loading, information recorded in pressure sensitive film was transformed to image file. After, by using image analysis software, the mean stress value and maximum stress value was calculated. RESULTS: The mean stress calculated at each osteotomy site is as follows; 3.89 MPa in the first pair; 4.55 MPa in the second pair; 4.62 MPa in the third pair; and 4.67 MPa in the fourth pair. In Group I, stress value was distributed evenly. But in group II, III, IV, the stress was concentrated at posteromedial area of the osteotomy site. CONCLUSION: The primary in the high tibial osteotomy was dependent more on the rigid fixation than on the continuity of the medial cortex. The pairs on which blade plate fixation was used more stable than the pairs on which staple fixation was used regardless of lateral translation.
Osteotomy* ; Tibia

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Cross-talk between Integrin and SMAD Signal Pathway in Articular Chondrocyte.

Soo Bong HAHN ; Min Sung PARK ; Yun Hee KIM ; Jin Woo LEE

Journal of Korean Orthopaedic Research Society.2007;10(2):65-75.

PURPOSE: Identifying the signal cross-talk between integrin signaling cascade and TGF-beta 1 signaling cascade in articular chondrocytes. MATERIALS AND METHODS: To analyze integrin or TGF-beta 1 mediated signaling pathways from extracellular stimuli, type II collagen was coated on the cell culture plate and TGF-beta 1 was added to cell culture media. Chondrocytes were cultured in the conditioned media with each or both stimuli. Altered activation of signaling proteins detected with western blot technique. RESULTS: More rapid attachment of cells was observed in the type II collagen coated group than non-coated group. The phosphorylated SMAD 2 and 3 were expressed in the type II collagen coated group and synergistically up-regulated phosphorylation in the co-treated group. The phosphorylated FAK at tyrosine 925 was activated by TGF-beta 1 treatment and synergistically up-regulated by both stimuli. But there was no meaningfully changed phosphorylation of extracellular signal regulated protein kinase (ERK) 1/2 and p38, as known downstream molecules of FAK cascade. CONCLUSION: This result means that SMAD 2, SMAD 3 and tyrosine 925 of FAK are involved in this signal cross-talking in articular chondrocytes.
Blotting, Western ; Cell Culture Techniques ; Chondrocytes* ; Collagen Type II ; Culture Media, Conditioned ; Phosphorylation ; Protein Kinases ; Signal Transduction* ; Transforming Growth Factor beta ; Tyrosine

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The Effect of Human Joint Fluid in the Surviviorship and Proliferation of Bone-Marrow Derived Progenitor Cell.

Kyu Bum SEO ; Dong Kee JUNG ; Sang Rim KIM ; Kwang Bok LEE

Journal of Korean Orthopaedic Research Society.2007;10(2):56-64.

PURPOSE: To investigate the effect of human joint fluid media on the survival and proliferation of bone marrow derived precusor cell, and to provide the basic data of the intraarticular injection with scaffold-free progenitor cell in advanced degenerative osteoarthritis (OA). MATERIALS AND METHODS: We obtained the joint fluid and bone marrow from 15 patients who had total knee arthroplasty due to degenerative OA, and isolated the mesenchymal progenitor cells (MPCs) from bone marrow by washing and ten times subculture. We devided the control and experiment groups according to the addition of joint fluid at various ratios (1/100, 1/10, 1, 10, 100, 1000 microliter), and statistically analyzed the numbers of mesenchymal progenitor cell proliferated according to the culture period. RESULTS: The experiments using joint fluid without centrifuge showed the increase of MPCs as the culture poriod was extended and was independent to the existence of fetal bovine serum, the dose dependent pattern in the increase of MPCs in proportion to the dose of joint fluid, and statistically significant increase MPCs in 10, 100, 1000 microliter of serum contained groups, 1000 microliter of serum-free groups (p=0.039, p=0.017, p=0.077, p=0.004). The experiments using joint fluid with centrifuge showed the increase of MPCs as the culture period was extended and was independent of fetal bovine serum, and the dose dependent pattern in the increase of MPCs in proportion to the dose of joint fluid, and statistically significant increase MPCs in 1000 microliter of both serum contained and serum-free groups (p=0.006, p=0.024). CONCLUSION: MSCs not only can survive, but also proliferate in human joint fluid. The rate of proliferaton is increased faster by the adding of joint fluid than only using common media in cell culture. And the experiment shows the dose dependent pattern in the increase of MPCs in proportion to the dose of joint fluid.
Arthroplasty ; Bone Marrow ; Cell Culture Techniques ; Humans* ; Injections, Intra-Articular ; Joints* ; Knee ; Mesenchymal Stromal Cells ; Osteoarthritis ; Stem Cells*

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