• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제36권5호
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• pp.392-401
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• 2010

Introduction: Diabetes mellitus, as a major health problem for the elderly has been shown to alter the properties of the bone and impair bone healing around a titanium implant in both humans and animals. The aim of this study was to examine the effect of adipose-derived stem cells on the healing process around a titanium implant in streptozotocin-induced diabetic rats. Materials and Methods: Thirteen rats were divided into two groups: adipose-derived stem cells injected group and a control group. A titanium screw implant (diameter: 2.0 mm, length: 3.5 mm) was placed into both tibia of 13 rats: 13 right tibia as the control group and 13 left tibia as the experimental group. The rats were sacrificed at different intervals (1, 2, and 4 weeks) after implantation for histopathology observations and immunohistochemistric analysis. Results: The histopathological findings revealed earlier new formed bone in the experimental group than the control group. In particular, at 1 week after implantation, the experimental group showed more newly formed bone and collagen around the implant than the control group. In immunohistochemistric analysis, osteoprotegerin (OPG) expression in the experimental group increased early compared to that of the control group until 2 weeks after implantation. However, after 2 weeks, OPG expression in the experimental group was similar to OPG expression in the control group. The receptor activator of nuclear factor ${\kappa}B$ ligand (RANKL) expression in the experimental group increased early compared to that of the control group, and then decreased at 2 weeks. After 2 weeks, the level of RANKL expression was similar in both groups. Conclusion: These results suggest that adipose-derived stem cells in implantation can promote bone healing around titanium, particularly in diabetes mellitus induced animals.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제37권5호
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• pp.386-395
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• 2011

Introduction: Osteoporosis is a major health problem in the elderly that involves changes in the properties of bone as well as impaired bone healing around a titanium implant in both humans and animals. This study examined effect of low intensity pulsed ultrasound (LIPUS) on the bone healing process around a titanium implant in osteoporosis-induced rats. Materials and Methods: Sixteen rats were divided into two groups. A control group with osteoporosis induced by removing both ovaries and an experimental group of rats that were applied with LIPUS after osteoporosis had been induced. A screw type titanium implant (diameter, 2.0 mm: length, 3.5 mm, Cowell-Medi, KOREA) was placed into the tibias of 16 rats. The control and experimental group contained 8 rats each. The rats were sacrificed at 1, 2, 4, and 8 weeks after implantation to examine the histopathology and immunochemistry. Results: The histopathology examination revealed earlier new bone formation in the experimental group than the control group. In particular, at 1 week after implantation, more new bone matrix and collagen were observed around the implant of the experimental group compared to the control group. Immunochemistry analysis showed that the level of OPG expression of the experimental group was higher in the early stages than in the control group. After 8 weeks, the levels of OPG expression were similar in both groups. The expression level of receptor activator of nuclear factor kB ligand (RANKL) was stronger in the experimental group than the control group. After 4 weeks, the level of RANKL expression was similar in both groups. Conclusion: These results suggest that the application of LIPUS to implantation can promote bone healing around titanium in osteoporosis animals.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제33권3호
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• pp.199-209
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• 2011

Purpose: Osteoporosis, is a major health problem for the elderly and post-menopausal women and shown to alter the properties of bone as well as impair bone healing around titanium implants in both human and animals. The objective of this study was to examine the effect of LIPUS with adipose-derived stem cells on the healing process around a titanium implant in rats with osteoporosis. Methods: Sixteen osteoporosis-induced rats were divided into two groups: an adipose-derived stem cell injected with Low Intensity Pulsed Ultrasound (LIPUS) application group and a control group. Titanium screw implants (diameter, 2.0 mm: length, 3.5 mm, Cowell Medi, Korea) were placed into both tibia of 16 rats, on 8 rats as the control group and the other 8 rats as the experimental group. Rats were sacrificed at different intervals from 1, 2, 4 and 8 weeks after implantation for histopathologic and immunohistochemical analyses. Results: Histopathological analysis revealed newly formed bone in experimental group earlier than that in control group. Especially at 1 week after implantation, more amounts of new bone matrix and collagen around the implant in the experimental group were seen compared with the control group. Immunohistochemical analysis showed that the levels of osteoprotegerin (OPG) expression in the experimental group were increased at early stages compared with that of control group until 2 weeks after implantation. But after 2 weeks, the expression level of OPG similar in both groups. The expression levels of receptor activator of nuclear factor kB ligand (RANKL) were stronger in the experimental group than the control group until 2 weeks after implantation. After 4 weeks, expression of RANKL in experimental group was similar to the control group. Conclusion: The results of this study suggest that LIPUS with Adipose-Derived Stem Cells in implantation could promote bone healing around titanium implants in rats with osteoporosis.

• 생명과학회지
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• 제25권2호
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• pp.223-230
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• 2015

RCAN1은 calcineurin을 억제하는 내인성 단백질로 calcineurin-NFATc1 신호전달 경로와 관련된 질환의 병인에 중요한 역할을 담당한다. 특히 RCAN1-4 아형 유전자의 경우 NFATc1 전사인자에 의해 조절된다. RANKL 자극은 calcineurin-NFATc1 경로로 파골세포 분화를 유도하는데, RCAN1과 파골세포의 분화에 관련된 연구는 보고 된 바 없다. 따라서 본 연구는 RANKL 처리에 의해 파골세포 분화가 유도될 때 RCAN1이 calcineurin-NFATc1 경로에 미치는 영향을 in vitro에서 조사했다. 마우스로부터 분리한 골수단핵세포에 RANKL을 처리하여 파골세포 분화를 유도했다. RANKL 처리 후 조사 대상 유전자의 mRNA 발현과 단백질 발현을 각각 RT-PCR과 Western blot로써 측정했다. 마우스 RCAN1-4 vector를 파골전구세포인 RAW 264.7 단핵세포주와 골수단핵세포에 형질도입(transfection)시켜 RCAN1-4 유전자의 과발현을 유도했다. 형질도입 후 파골세포 분화의 형태적 변화는 TRAP 염색을 통해 관찰했다. RANKL 처리 후 NFATc1, calcineurin, RCAN1-4 mRNA 발현은 크게 증가했다. 단백질 발현의 경우 NFATc1과 RCAN1은 증가했으나 calcineurin은 대조군과 차이가 없었다. RCAN1-4 유전자의 과발현 유도 시 RCAN1-4 mRNA는 크게 증가되었으나 RCAN1 단백질 발현은 증가되지 않았다. 특히 RANKL 존재 시 RCAN1 유전자를 knock-down시켜도 RCAN1 발현은 정상적으로 유지되었다. 한편, NFATc1 발현은 과발현 유도시 감소했고 knock-down 유도 시 증가하는 경향을 보였다. RCAN1-4 유전자 과발현을 유도한 골수단핵세포에서 배양 5일 후 파골세포 분화는 대조군과 차이가 없었다. 이러한 결과는 RANKL에 의한 파골세포 분화 시 RCAN1이 calcineurin-NFATc1 경로를 통해 파골세포 분화에 미치는 영향은 제한적일 것으로 사료된다.

• 한국식품영양과학회지
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• 제37권8호
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• pp.992-997
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• 2008

본 연구는 MC3T3-E1 조골전구세포에 실크피브로인 처리 시 파골세포분화 관련 국소인자 중 파골세포로의 분화를 강력히 유도하는 IL-$1{\beta}$의 발현 및 파골세포로의 분화억제에 관여하는 국소인자인 OPG의 발현변화를 조골세포 조건 배양액에서 살펴보았으며, 이 조골세포 조건배양액을 RAW264.7 파골전구세포에 처리 시 파골세포로의 분화를 억제하는지를 알아보았다. 조골세포 조건배양액에서 OPG의 발현은 농도별($0.001\;mg/mL{\sim}0.1\;mg/mL$) 실크피브로인 처리 시 $86%{\sim}100%$로 양성대조군(+C)과 유의적 차이가 없었다. 그러나 IL-$1{\beta}$의 발현은 0.1 mg/mL 실크피브로인 처리 시 양성대조군(+C)의 1.8% 수준으로 그 발현을 현저히 억제 하였다. 조골세포조건배양액을 처리한 RAW264.7 세포의 파골세포로의 분화정도는 농도 의존적으로 현저히 감소 하였다. 특히, 0.1 mg/mL의 실크피브로인을 처리한 조골세포 조건배양액 처리 시 파골세포로 분화하지 않은 음성대조군(-RANKL) 수준으로 파골세포 분화를 현저히 억제한 것은 OPG의 발현 증가가 아닌 IL-$1{\beta}$의 발현 억제에 의한 것으로 사료되며, 추후 연구를 통해 관련 기전의 연구가 필요할 것으로 생각되어진다.

• 생명과학회지
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• 제30권11호
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• pp.983-989
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• 2020

파골세포에 의한 뼈의 재흡수와 조골세포에 의한 뼈 형성의 균형은 뼈 건강에 매우 중요한 요소이다. 따라서 이들의 불균형은 골다공증, 골연화증 및 골석화증과 같은 다양한 골 질환을 유발할 수 있다. 특히 파골세포의 기능이 비정상적으로 항진된 경우 골 파괴가 증가되어 골다공증이 야기되며 이런 현상은 류마티스 관절염 같은 염증성 질환의 골 소실과 밀접한 연관이 있다. 그러나 현재 사용중인 골 흡수 억제제는 장기간 사용시 부작용이 보고되고 있어 천연물 소재를 기반으로한 새로운 골 흡수 억제제 개발이 요구되고 있다. 따라서 본 연구에서는 갈색거저리 유충 에탄올 추출물이 RANKL에 의해 유도되는 파골세포 분화에 미치는 영향을 확인하고 그 작용기작을 구명하고자 하였다. 갈색거저리 유충 에탄올 추출물이 파골세포 분화에 미치는 영향을 조사하기 위하여 RAW264.7 세포에 RANKL을 단독 처리 및 추출물과 함께 5일간 처리한 후 TRAP 활성을 비교하였다. 그 결과 RANKL에 의해 증가한 파골세포 분화는 갈색거저리 유충 추출물 2 mg/ml 농도까지 세포독성 없이 농도 의존적으로 감소하는 것을 확인할 수 있었다. 또한 갈색거저리 추출물이 TRAP, NFATc1 및 CtsK와 같은 파골세포 분화마커 유전자 및 단백질 발현량에 미치는 변화를 확인한 결과, RANKL 처리에 의해 현저히 증가한 이들 유전자가 갈색거저리 추출물에 의해 현저하게 감소하는 것을 확인하였다. 또한 갈색거저리 추출물의 파골세포 분화 억제작용기작을 확인한 결과 mitogen activated protein kinases (MAPKs)중 p38의 신호 전달 억제 기작을 통하여 파골세포 분화가 억제됨을 확인 할 수 있었다. 이러한 결과로 보아 갈색거저리 유충 에탄올 추출물 및 그 생리활성 물질들은 골다공증과 같은 골 질환 치료 및 예방을 위한 잠재적 기능성 소재로 사용될 수 있음을 확인하였다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제37권4호
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• pp.301-311
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• 2011

Introduction: This study examined the effect of the application of low intensity pulsed ultrasound on bone healing after an injection of adipose tissue-derived stem cells (ADSCs) during the implantation of a titanium implant in the tibia of diabetes-induced rats. Materials and Methods: Twelve Sprague-Dawely rats were used. After inducing diabetes, the ADSCs were injected into the hole for the implant. Customized screw type implants, 2.0 mm in diameter and 3.5 mm in length, were implanted in both the tibia of the diabetes-induced rats. After implantation, LIPUS was applied with parameters of 3 MHz, 40 mW/$cm^2$, and 10 minutes for 7 days to the left tibiae (experimental group) of the diabetesinduced rats. The right tibiae in each rat were used in the control group. At 1, 2 and 4 week rats were sacrificed, and the bone tissues of both tibia were harvested. The bone tissues of the three rats in each week were used for bone-to-implant contact (BIC) and bone area (BA) analyses and the bone tissues of one rat were used to make sagittal serial sections. Results: In histomorphometric analyses, the BIC in the experimental and control group were respectively, $39.00{\pm}18.17%$ and $42.87{\pm}9.27%$ at 1 week, $43.74{\pm}6.83%$ and $32.27{\pm}6.00%$ at 2 weeks, and $32.62{\pm}11.02%$ and $47.10{\pm}9.77%$ at 4 weeks. The BA in experimental and control group were respectively, $37.28{\pm}3.68%$ and $31.90{\pm}2.84%$ at 1 week, $20.62{\pm}2.47%$ and $15.64{\pm}2.69%$ at 2 weeks, and $11.37{\pm}4.54%$ and $17.69{\pm}8.77%$ at 4 weeks. In immunohistochemistry analyses, Osteoprotegerin expression was strong at 1 and 2 weeks in the experimental group than the control group. Receptor activator of nuclear factor kB ligand expression showed similar staining at each week in the experimental and control group. Conclusion: These results suggest that the application of low intensity pulsed ultrasound after an injection of adipose tissue-derived stem cells during the implantation of titanium implants in the tibia of diabetes-induced rats provided some positive effect on bone regeneration at the early stage after implantation. On the other hand, this method is unable to increase the level of osseointegration and bone regeneration of the implant in an uncontrolled diabetic patient.