• Maxillofacial Plastic and Reconstructive Surgery
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• 제39권
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• pp.8.1-8.8
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• 2017

Background: For an effective bone graft for reconstruction of the maxillofacial region, an adequate vascular network will be required to supply blood, osteoprogenitor cells, and growth factors. We previously reported that the secretomes of bone marrow-derived mesenchymal stem cells (MSC-CM) contain numerous growth factors such as insulin-like growth factor (IGF)-1, transforming growth factor $(TGF)-{\beta}1$, and vascular endothelial growth factor (VEGF), which can affect the cellular characteristics and behavior of regenerating bone cells. We hypothesized that angiogenesis is an important step for bone regeneration, and VEGF is one of the crucial factors in MSC-CM that would enhance its osteogenic potential. In the present study, we focused on VEGF in MSC-CM and evaluated the angiogenic and osteogenic potentials of MSC-CM for bone regeneration. Methods: Cytokines in MSC-CM were measured by enzyme-linked immunosorbent assay (ELISA). Human umbilical vein endothelial cells (HUVECs) were cultured with MSC-CM or MSC-CM with anti-VEGF antibody (MSC-CM + anti-VEGF) for neutralization, and tube formation was evaluated. For the evaluation of bone and blood vessel formation with micro-computed tomography (micro-CT) and for the histological and immunohistochemical analyses, a rat calvarial bone defect model was used. Results: The concentrations of IGF-1, VEGF, and $TGF-{\beta}1$ in MSC-CM were $1515.6{\pm}211.8pg/mL$, $465.8{\pm}108.8pg/mL$, and $339.8{\pm}14.4pg/mL$, respectively. Tube formation of HUVECs, bone formation, and blood vessel formation were increased in the MSC-CM group but decreased in the MSC-CM + anti-VEGF group. Histological findings suggested that new bone formation in the entire defect was observed in the MSC-CM group although it was decreased in the MSC-CM + anti-VEGF group. Immunohistochemistry indicated that angiogenesis and migration of endogenous stem cells were much more abundant in the MSC-CM group than in the MSC-CM + anti-VEGF group. Conclusions: VEGF is considered a crucial factor in MSC-CM, and MSC-CM is proposed to be an adequate therapeutic agent for bone regeneration with angiogenesis.

• Journal of Veterinary Science
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• 제24권6호
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• pp.83.1-83.12
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• 2023

Background: Ellipticine (Ellip.) was recently reported to have beneficial effects on the differentiation of adipose-derived stem cells into mature chondrocyte-like cells. On the other hand, no practical results have been derived from the transplantation of bone marrow stem cells (BMSCs) in a rabbit osteoarthritis (OA) model. Objectives: This study examined whether autologous BMSCs incubated with ellipticine (Ellip.+BMSCs) could regenerate articular cartilage in rabbit OA, a model similar to degenerative arthritis in human beings. Methods: A portion of rabbit articular cartilage was surgically removed, and Ellip.+BMSCs were transplanted into the lesion area. After two and four weeks of treatment, the serum levels of proinflammatory cytokines, i.e., tumor necrosis factor α (TNF-α) and prostaglandin E2 (PGE2), were analyzed, while macroscopic and micro-computed tomography (CT) evaluations were conducted to determine the intensity of cartilage degeneration. Furthermore, immuno-blotting was performed to evaluate the mitogen-activated protein kinases, PI3K/Akt, and nuclear factor-κB (NF-κB) signaling in rabbit OA models. Histological staining was used to confirm the change in the pattern of collagen and proteoglycan in the articular cartilage matrix. Results: The transplantation of Ellip.+BMSCs elicited a chondroprotective effect by reducing the inflammatory factors (TNF-α, PGE2) in a time-dependent manner. Macroscopic observations, micro-CT, and histological staining revealed articular cartilage regeneration with the downregulation of matrix-metallo proteinases (MMPs), preventing articular cartilage degradation. Furthermore, histological observations confirmed a significant boost in the production of chondrocytes, collagen, and proteoglycan compared to the control group. Western blotting data revealed the downregulation of the p38, PI3K-Akt, and NF-κB inflammatory pathways to attenuate inflammation. Conclusions: The transplantation of Ellip.+BMSCs normalized the OA condition by boosting the recovery of degenerated articular cartilage and inhibiting the catabolic signaling pathway.

• 한국임상수의학회지
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• 제27권4호
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• pp.325-329
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• 2010

본 연구에서는 hydroxyapatite/poly $\varepsilon$-caprolactone composite (HA/PCL) 지지체와 matrigel을 랫드의 두개골 결손부 모델에 함께 이식 시의 골형성 정도를 평가하였다. 두개골결손부는 Sprague Dawley rat (n = 18)에서 수술적으로 형성하였으며 실험군은 Matrigel과 함께 HA/PCL 지지체를 이식한 군(M-HA/PCL group, n = 6)과 HA/PCL 지지체단독이식군(HA/PCL group, n = 6)으로 나누었고 대조군(CD group, n = 6)에는 아무 것도 이식하지 않았다. 수술 4주 후, 골형성은 방사선촬영, micro CT 및 조직검사를 통해 평가되었다. 방사선상에서 CD군의 골형성은 관찰되지 않았으나 HA/PCL과 M-HA/PCL군에서는 관찰되었고 골과 유사한 방사선비투과성이 M-HA/PCL군에서 더 많이 관찰되었다. Micro CT 평가에서 골부피는 HA/PCL군보다 M-HA/PCL군에서 더 높았으나 두 군 사이의 유의적 차이는 관찰할 수 없었다. 그러나 골밀도에서는 HA/PCL군보다 M-HA/PCL군이 더 유의적으로 높음을 확인할 수 있었다(p < 0.05). 조직학적 검사에서는 CD군에서 새로운 골은 원래 존재하던 골로부터만 형성되었으며 두개골결손부 내의 골형성은 보이지 않았다. HA/PCL군에서 새로운 골형성은 원래 존재하던 골로부터만 유래되었으나 M-HA/PCL군은 가장 많은 골형성을 보여주었으며 새로운 골이 원래 존재하던 골과 HA/PCL지지체 주변에서도 관찰되었다. 이러한 결과로 미루어볼 때 HA/PCL 지지체와 matrigel을 함께 사용하는 것이 골의 임계결손부에서 골형성을 증대시키는 효과적인 방법이 될 수 있을 것으로 생각된다.

• 대한한의학회지
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• 제34권1호
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• pp.116-135
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• 2013

Objectives: This study was intended to clarify how Jakyakkamchobuja-tang (hereinafter referred to JKBT) affects mice of C57BL/10 whose osteoarthritis was induced by papain. Methods: Osteoarthritis was induced in mice by injecting papain in the knee joint. Mice were divided into 4 groups (n=6). The normal group were not treated at all whereas the control group (OAC-control) were induced for osteoarthritis by papain and oral medicated with 200 ul of physiological saline per day. The positive comparison group (OAC-$Joins^{(R)}$) were injected with papain and after 7 days, 100 mg/kg of $Joins^{(R)}$ were medicated with 200 ul of physiological saline mixed. The experimental group (OAC-JKBT) were injected with papain and after 7 days were medicated with 400 mg/kg of JKBT mixed with 200 ul of physiological saline. OAC-$Joins^{(R)}$ and OAC-JKBT were oral medicated for each substance for a total of 4 weeks, once per day. After experiments (from 1 week after injection of papain to 4 weeks elapsed), the function of liver and kidney, inflammation cytokine values within serum, degree of revelation for inflammation cytokine genes, immune cells within blood, metabolism of arachidonic acid and amount of cartilage were measured and histopathological variations for knee joint structures were observed. Results: Functions of liver and kidney were not affected. IL-$1{\beta}$ (interleukin-$1{\beta}$), MCP-1 (monocyte chemoattractant protein-1) and TNF-${\alpha}$ (tumor necrosis factor-${\alpha}$) were significantly reduced and IL-6 (interleukin-6) was also reduced but not significantly. After analyzing inflammation cytokine in joints with mRNA (messenger ribonucleic acid), revelation of IL-6, TNF-${\alpha}$, COX-2 (cyclooxygenase-2) and iNOS-II (inducible nitric oxide synthase-II) were all significantly reduced. Revelation of IL-$1{\beta}$ gene was also reduced but not significantly. Neutrophil for WBC (white blood cell) within serum was significantly reduced; monocyte was also reduced but not significantly. PGE2 (prostaglandin E2), TXB2 (thromboxane B2) were significantly reduced and LTB4 (leukotriene B4) was also reduced but not significantly. Destruction of cartilage on micro CT (computed tomography)-arthrography was reduced but had no significant differences. In terms of histopathology, infiltration of inflammation, proliferation of synovial membrane, subsidence of cartilage and bone due to penetration of excessive formation of synovial cell and destruction of cartilage were small (H&E (hematoxylin and eosin), safranine O staining). Conclusions: Based on these results, Jakyakkamchobuja-tang (JKBT) is believed to be useful for suppressing the progress of osteoarthritis and its treatments because of its anti-inflammatory effects and alleviation of pain with histopathological effective efficacy.

• International Journal of Industrial Entomology and Biomaterials
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• 제30권1호
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• pp.17-25
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• 2015

This study aimed to investigate the effects of silk fibroin on bone metabolism in ovariectomized rats. A total of 30 Sprague-Dawley rats were randomized into sham-operated (SHAM), ovariectomized control (OVX), alendronate (OVX+ALEN, 10 mg/kg body weight/d), low silk fibroin (OVX+SF100, 100 mg/kg body weight/d), and high silk fibroin (OVX+SF300, 300 mg/kg body weight/d) groups. All the rats were fed by gavage for 12 wk. At the end of 12 wk, blood and urine were collected for analysis of bone turnover markers, and bone mineral density (BMD) was measured by micro-computed tomography. The results show that the OVX group (p < 0.05) displayed the highest mean body weight gain. Among the five groups, serum levels of bone alkaline phosphatase (ALP) and urine levels of deoxypyridinoline (DPD) were highest in the OVX group (p < 0.05). Bone ALP levels in the ALEN group were significantly lower than that of the silk-treated groups. On the other hand, DPD levels were not significantly different between the ALEN and silk-fibroin-treated groups (p < 0.05). The trabecular BMD was significantly higher in the ALEN and silk-treated groups compared to the OVX group (p < 0.05). In conclusion, this study showed that silk fibroin has similar effects as alendronate, which is used in osteoporosis medication. Therefore silk fibroin might be a new candidate for the prevention and treatment of osteoporosis in patients.

• 대한기계학회논문집B
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• 제36권11호
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• pp.1081-1090
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• 2012

외상성 관절염 발생시 슬관절에서의 해면골 미세구조 변화 특성에 대한 연구는 부족한 실정이다. 따라서 본 연구에서는 생체 내 미세 단층 촬영을 통해, 소동물(SD rat; 10 마리)의 경골 골단 해면골 미세구조 변화패턴을 연속적으로 추적 관찰하여 정량적으로 분석하였다. 외상성 관절염 군(5 마리)에는 전방십자인대절제술을 실시하였고, 정상군(5 마리)에는 아무런 외과적 처치를 가하지 않았다. 외상성 관절염군은 정상군과 비교시 수술후 8 주까지 약 4-16%의 유의한 골 미세구조 차이를 나타내었다(P<0.05). 동일 기간 동안 골 밀집 정도 및 골 형성 정도는 5-15% 정도로 유의한 차이를 나타내었다(P<0.05). 본 연구는 외상성 관절염 발생에 의한 경골 골단 해면골의 골 미세구조 변화 특성 기준 정립 및 외상성 관절염의 이해와 치료에 도움이 될 것으로 기대된다.

• Journal of Korean Dental Science
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• 제8권2호
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• pp.65-73
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• 2015

Purpose: Parathyroid hormone (PTH) therapy has drawn attention, as an alternative to anti-resorptive drugs since PTH accelerates bone density by anabolic action. The purpose of this study was to identify the effect of intermittent PTH administration on jaw bones of rat undergone bilateral ovariectomy. Materials and Methods: Nine female Sprague-Dawley rats were divided into three groups. PTH group was ovariectomized (OVX) to induce osteoporosis and PTH $30{\mu}g/kg$ was administered 1 week after the surgery. In OVX group, ovariectomy was performed and only vehicle was administered by subcutaneous injection 3 times per week. Control group was subjected to sham surgery. The animals were sacrificed 8 weeks after the surgery and specimens were obtained from ilium and upper and lower jaw bones. Histological investigation was carried out by using an optical microscope and micro-computed tomography was taken to examine structural property changes in each bone sample. Result: In the ilium, the bone volume ratio (bone volume/total volume, BV/TV) of PTH, OVX and control groups was $53.75%{\pm}7.57%$, $50.61%{\pm}12.89%$, $76.20%{\pm}5.92%$ (P=0.061) and bone mineral density (BMD) was $1.12{\pm}0.09$, $0.88{\pm}0.48$, $1.38{\pm}0.07g/cm^3$ (P=0.061). In the mandible, BV/TV of PTH, OVX and control groups was $64.60%{\pm}12.17%$, $58.26%{\pm}9.63%$, $67.54%{\pm}14.74%$(P=0.670) and BMD was $1.21{\pm}0.17$, $1.19{\pm}0.13$, $1.27{\pm}0.18g/cm^3$ (P=0.587). In the maxilla, BV/TV of PTH, OVX and control groups was $61.19%{\pm}8.92%$, $52.50%{\pm}11.22%$, $64.60%{\pm}12.17%$ (P=0.430) and BMD was $1.20{\pm}0.11$, $1.11{\pm}0.16$, $1.21{\pm}0.17g/cm^3$ (P=0.561). No statistically significant difference was found in any variables in all groups. Histological observation revealed that the ilium in OVX group demonstrated sparsely formed trabecular bones compared with other groups. However, upper and lower trabecular bones did not present significant differences. Conclusion: Intermittent administration of PTH appears to affect the microstructure of rat jaw bones, but statistical significance was not found. However, the measurements in this study partly implicated the possible anabolic effect of PTH in vivo.

• Journal of Periodontal and Implant Science
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• 제49권6호
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• pp.382-396
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• 2019

Purpose: The purpose of this study was to elucidate the efficacy and safety of carbonate apatite (CO₃Ap) granules in 2-stage sinus floor augmentation through the radiographic and histomorphometric assessment of bone biopsy specimens. Methods: Two-stage sinus floor augmentation was performed on 13 patients with a total of 17 implants. Radiographic assessment using panoramic radiographs was performed immediately after augmentation and was also performed 2 additional times, at 7±2 months and 18±2 months post-augmentation, respectively. Bone biopsy specimens taken from planned implant placement sites underwent micro-computed tomography, after which histological sections were prepared. Results: Postoperative healing of the sinus floor augmentation was uneventful in all cases. The mean preoperative residual bone height was 3.5±1.3 mm, and this was increased to 13.3±1.7 mm by augmentation with the CO₃Ap granules. The mean height of the augmented site had decreased to 10.7±1.9 mm by 7±2 months after augmentation; however, implants with lengths in the range of 6.5 to 11.5 mm could still be placed. The mean height of the augmented site had decreased to 9.6±1.4 mm by 18±2 months post-augmentation. No implant failure or complications were observed. Few inflammatory cells or foreign body giant cells were observed in the bone biopsy specimens. Although there were individual differences in the amount of new bone detected, new bone was observed to be in direct contact with the CO₃Ap granules in all cases, without an intermediate layer of fibrous tissue. The amounts of bone and residual CO₃Ap were 33.8%±15.1% and 15.3%±11.9%, respectively. Conclusions: In this first demonstration, low-crystalline CO₃Ap granules showed excellent biocompatibility, and bone biopsy showed them to be replaced with bone in humans. CO₃Ap granules are a useful and safe bone substitute for two-stage sinus floor augmentation.

• Journal of Korean Neurosurgical Society
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• 제63권4호
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• pp.433-443
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• 2020

Objective : Osteoporosis is a disease of unbalanced bone metabolism that results in low bone mineral density with increased bone fragility and propensity for fractures. The increased rate of bone fracture due to osteoporosis places a significant burden on public health care expenditures. Therefore, numerous studies have been designed and performed to identify the drugs or health foods that can improve the bone quality or quantity. This study was designed to evaluate and analyze the therapeutic effects of rutin on histomorphometric values of the spine and femur in an osteoporotic mouse model induced by bilateral ovariectomy. Methods : Thirty female ICR mice (8 weeks old) underwent either a sham operation (only abdominal incision, sham group, n=10) or bilateral ovariectomy (n=20). The ovariectomized (OVX) animals were randomly divided into two groups : untreated OVX group (OVX-C, n=10), or rutin-administered group (OVX-R, n=10). The OVX-C group received weight-adjusted doses of saline vehicle and the OVX-R group received 50 mg/kg of rutin intraperitoneally, starting 1 day after surgery. At 4 and 8 weeks after surgery, serum estrogen, osteocalcin, alkaline phosphatase (ALP), and the telopeptide fragment of type I collagen C-terminus (CTX-1) were analyzed. Interleukin (IL)-1β, IL-6, IL-10, and tumor necrosis factor (TNF)-α were also analyzed. Bone histomorphometric parameters of the 4th lumbar vertebra and femur were determined by micro-computed tomography. Results : In OVX-C group, ALP, osteocalcin, CTX-1, IL-1β, IL-6, and TNF-α levels were significantly increased at 4 and 8 weeks compared to sham operation group. Rutin administration after OVX statistically significantly reduced ALP, CTX-1, IL-1β, IL-6, and TNF-α levels at 4 and 8 weeks. Rutin administration also improves bone histomorphometric parameters including trabecular bone volume fraction, trabecular thickness, and trabecular number. Trabecular separation was also decreased in OVX-R group compared to OVX-C group. Conclusion : The present study demonstrated that rutin has therapeutic effects on improving bone histomorphometric values in an OVX mouse model. The improvement in histomorphometric values may be associated with the reduction of osteoclastic activity via inhibition of IL-1β, IL-6, and TNF-α. In future studies, the mechanism for the effect of rutin on osteoporosis should be demonstrated more clearly to use rutin in human osteoporosis.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제34권6호
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• pp.391-397
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• 2012

Purpose: The purpose of this study was to evaluate the effectiveness of the platelet-rich fibrin (PRF) used in combination with the porcine cancellous bone as a scaffold, in promoting bone regeneration in the bone defects ofthe rabbit calvaria. Methods: Ten rabbits were used in the study. Three round-shaped defects (diameter 8.0 mm) were created in the rabbit calvaria and were filled with nothing (control group), porcine cancellousbone (Experimental Group 1, porcine bone) and PRF-mixed porcine cancellous bone (Experimental Group 2). TS-GBB is a xenogenic bone-substitute product comprised of a high heat-treated mineralized porcine cancellous bone. Animals were sacrificed at 6 weeks and 12 weeks for the histological and radiographic evaluations. Results: In the micro computed tomography and histological results, the experimental groups 1 and 2 showed more bone formation, remodeling, and calcification than the control group. The new bone formation ratio showed theGroup 2 to be larger than Group 1 at6 and 12 weeks. However, there was no significant difference between the experimental groups 1 and 2 in the new bone formation area, at the 6 and 12 weeks (P>0.05). Conclusion: The PRF-mixed group showed more bone formation than the porcine cancellousbonegroup (TS-GBB), butthere was a no significant difference. The PRF may not lead to enhanced bone healing when grafted with the porcine cancellous bone.