• YAKHAK HOEJI
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• v.49 no.6
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• pp.495-504
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• 2005

In our study, osteoporosis was induced by ovariectomized in female Sprague-Daweley rats, and the prevention and treatment efficacy of the climacteric disease the postmenopausal Type I pattern was examined by using the experimental substance soybeans and arrowroot. After ovariectomy; the amount of estrogen was found to be reduced by 21.37$\%$ and it showed the tendency to be increased by 4.49$\%$ in T1 (extraction of soybeans group) and 7.62$\%$ in T2 (extraction of ferment soybeans group). The change of bone density, 14 weeks after ovariectomy in the femur, was decreased by 9.33$\%$ in N.C. group, and 5.46$\%$ in T3 (extraction of ferment arrowroot group). As pathological findings, the trabeculae of femur were detected to be abundant as normal and the frequency of the appearance of osteoclasts was very minimal. In the T3, the frequency of the appearance of the trabeculae of femur was increased to the level that could be distinguished. In regard to the accumulation of lipid in the bone marrow, the accumulation of lipid in the bone marrow was low as normal. However, in T3, the decrease to the level that could be distinguished was detected. In addition, in regard to the pathological findings of the uterus, normal uterine tissue findings were detected. In the T3, the minimal atrophy findings were detected. In conclusion, when the T3 was orally administered continuously for 14 weeks, it was thought that a certain proportion of the hormonal balance was maintained that functioned as a substance interfering the accumulation of fat, and it was considered to be of help in the treatment of not only osteoporosis Type I, but also for the prevention and treatment of various endocrinal diseases.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.29 no.6
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• pp.397-406
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• 2003

The purpose of this study was to observe the titanium implant osseointegration in the osteoporosis-induced animal model. Seventy rats, 11 weeks of age, were divided into two groups : an ovariectomized group and a control group. Titanium screw implants(diameter, 2.0mm; length, 3.5mm) were placed into left tibias of 70 rats, 35 in the control group and 35 in the experimental group. The rats were sacrificed at different time interval (1, 2, 3, 4, 6, 8, and 12 weeks after implantation) for histopathologic observation, histomorphometric analysis and immunohistochemistry with fibronectin and CD34 antibody. The results obtained from this study were as follows: 1. Histopathologically findings, newly formed bone was seen at 3 weeks and became lamellar bone at 8 weeks, and mature trabecullar bone was seen at 12 weeks control group. In experimental group, thickness of regenerated bone increased till 8 weeks gradually and mature trabecullar bone was seen at 12 weeks. 2. By histomorphometric analysis, marrow bone density and contact ratio of marrow bone to implant decreased significantly from 8 to 12 weeks in experimental group compared to control group and also total bone to implant contact ratio decreased significantly from 8 to 12 weeks in experimental group. 3. Fibronectin immunoreactivity was strong at 3 weeks control group and reduced after 8weeks gradually. But it was continuously strong from 3 to 8 weeks in experimental group. 4. CD34 immunoreactivity was very strong in the newly formed osteoblasts from 3 to 8 weeks control group. But it reacted minimally later. While in experimental group, it reacted continuously strong from 3 to 12 weeks. The results of this study suggest that osteoporosis is not an absolute contraindication to dental implantation if sufficient period suggested after fixture installation till second stage surgery.

• Journal of Korean Neurosurgical Society
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• v.56 no.5
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• pp.375-382
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• 2014

Objective : To assess the effect of bone marrow mononuclear cells (BMMNCs) transplantation in the expression of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) in spinal cord injury (SCI) in rats. Methods : BMMNCs were isolated from tibia and femur by a density gradient centrifugation. After establishment of acute transection SCI, rats were divided into experiment (BMMNCs), experiment control (0.1 M PBS infused) and sham surgery groups (laminectomy without any SCI). Locomotor function was assessed weekly for 5 weeks post-injury using BBB locomotor score and urinary bladder function daily for 4 weeks post-injury. Activity of NF-${\kappa}B$ in spinal cord was assessed by immunohistochemistry and reverse transcriptase polymerase chain reaction. Results : At each time point post-injury, sham surgery group had significantly higher Basso, Beattie, Bresnahan locomotor and urinary bladder function scores than experiment and experiment control group (p<0.05). At subsequent time interval there were gradual improvement in both experiment and experiment control group, but experiment group had higher score in comparison to experiment control group (p<0.05). Comparisons were also made for expression of activated NF-${\kappa}B$ positive cells and level of NF-${\kappa}B$ messenger RNA in spinal cord at various time points between the groups. Activated NF-${\kappa}B$ immunoreactivity and level of NF-${\kappa}B$ mRNA expression were significantly higher in control group in comparison to experiment and sham surgery group (p<0.05). Conclusion : BMMNCs transplantation attenuates the expression of NF-${\kappa}B$ in injured spinal cord tissue and thus helps in recovery of neurological function in rat models with SCI.

• Journal of Embryo Transfer
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• v.27 no.3
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• pp.183-187
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• 2012

Adult stem cell transplantation has been increased every year, because of the lack of organ donors for regenerative medicine. Therefore, development of reliable and safety cryopreservation and bio-baking method for stem cell therapy is urgently needed. The present study investigated safety of dimethyl sulfoxide (DMSO) such as common cryoprotectant on porcine bone marrow derived mesenchymal stem cells (pBM-MSCs) by evaluating the activation of Caspase-3 and -7, apoptosis related important signal pathway. pBM-MSCs used for the present study were isolated density gradient method by Ficoll-Paque Plus and cultured in A-DMEM supplemented 10% FBS at $38.5^{\circ}C$ in 5% $CO_2$ incubator. pBM-MSCs were cryopreserved in A-DMEM supplemented either with 5%, 10% or 20% DMSO by cooling rate at $-1^{\circ}C$/min in a Kryo 360 (planner 300, Middlesex, UK) and kept into $LN_2$. Survival rate of cells after thawing did not differ between 5% and 10% DMSO but was lowest in 20% DMSO by 0.4% trypan blue exclusion. Activation of Caspase-3 and -7 by Vybrant FAM Caspase-3 and -7 Assay Assay Kit (Molecular probes, Inc.OR, USA) was analyzed with a flow cytometer. Both of cryopreserved and control groups (fresh pBM-MSCs) were observed after the activation of Caspase-3 and -7. The activation did not differ between 5% and 10% DMSO, but was observed highest in 20% DMSO. Therefore 5% DMSO can be possibly used for cell cryopreservation instead of 10% DMSO.

• Genomics & Informatics
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• v.2 no.2
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• pp.75-80
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• 2004

In a variety of animal species, the perinatal exposure of experimental animals to the 2,3,7,8-tetrachlorodibenzo­p-dioxin (TCDD) leads to the immune dysfunction, which is more severe and persistent than that caused by adult exposure. We report here the changes of gene expression and the identification of the marker-genes representing the dioxin exposure. The expressions of the transcripts were analyzed using the 11 K oligonucleotide­microarray from the bone marrow cells of male C57BL/6J mice after an intraperitoneal injection of $1{\mu}g$ TCDD/kg body weight at various time intervals: gestational 6.5 day(G6.5), 13.5 day(G13.5), 18.5 day(G18.5), and postnatal 3 (P3W)and 6 week (P6W). The type of self-organizing maps(SOM) representing the specific exposure dioxin could be identified as follows; G6.5D(C14), G13.5D(C0, C5, C10, C18), G18.5D(7): P3W(C2, C21), and P6W(C4, C15, C20). The candidate marker-genes were restricted to the transcripts, which could be consistently expressed greater than $\pm$2-fold in three experiments. The resulting candidates were 85 genes, the characteristics of that were involved in cell physiology and cell functions such as cell proliferation and immune function. We identified the biomarker-genes for dioxin exposure: smc -like 2 from SOM C14 for the dioxin exposure at G6.5D, focal adhesion kinase and 6 other genes from C0, and protein tyrosine phosphatase 4a2 and 3 other genes from C5 for G13.5D, platelet factor 4 from C7 for G18.5D, fos from C2 for P3W.

• Polymer(Korea)
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• v.28 no.3
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• pp.218-224
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• 2004

The adhesion and proliferation of mammalian cells on polymeric biomaterials depend on the surface characteristics such as wettability, chemistry, charge and roughness. In order to recognize the correlation between the adhesion and proliferation of human bone marrow derived stem cells (BMSCs) and surface property, radio frequency generated plasma treatment on low density polyethylene (LDPE) has been carried out. The modified LDPE surfaces were characterized by measuring the static water contact angle. The adhesion and proliferation of cells on LDPE films were characterized by cell counting and reverse transcription-polymerase chain reaction (RT-PCR). The water contact angle of the film surface decreased with plasma treatment time. Proto-oncogenes (c-myc, c-fos) and tumor suppressor gene (p153) showed maximum expression with contact angle of 60 ∼ 70$^{\circ}$ range of LDPE film. By cell counting, we confirmed that the rate of cell proliferation appeared the higher on the film surface of the contact angle of 60∼70$^{\circ}$ We concluded that the surface wettability is an important role for the growth and differentiation of BMSCs.

• BMB Reports
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• v.52 no.11
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• pp.659-664
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• 2019

Vav1 is a Rho/Rac guanine nucleotide exchange factor primarily expressed in hematopoietic cells. In this study, we investigated the potential role of Vav1 in osteoclast (OC) differentiation by comparing the ability of bone marrow mononuclear cells (BMMCs) obtained from Vav1-deficient ($Vav1^{-/-}$) and wild-type (WT) mice to differentiate into mature OCs upon stimulation with macrophage colony stimulating factor and receptor activator of nuclear kappa B ligand in vitro. Our results suggested that Vav1 deficiency promoted the differentiation of BMMCs into OCs, as indicated by the increased expression of tartrate-resistant acid phosphatase, cathepsin K, and calcitonin receptor. Therefore, Vav1 may play a negative role in OC differentiation. This hypothesis was supported by the observation of more OCs in the femurs of $Vav1^{-/-}$ mice than in WT mice. Furthermore, the bone status of $Vav1^{-/-}$ mice was analyzed in situ and the femurs of $Vav1^{-/-}$ mice appeared abnormal, with poor bone density and fewer number of trabeculae. In addition, Vav1-deficient OCs showed stronger adhesion to vitronectin, an ${\alpha}_v{\beta}_3$ integrin ligand important in bone resorption. Thus, Vav1 may inhibit OC differentiation and protect against bone resorption.

• Archives of Plastic Surgery
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• v.38 no.4
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• pp.438-444
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• 2011

Purpose: Adipose-derived stromal cells (ASCs) are readily harvested from lipoaspirated tissue or subcutaneous adipose tissue fragments. The stromal vascular fraction (SVF) is a heterogeneous set of cell populations that surround and support adipose tissue, which includes the stromal cells, ASCs, that have the ability to differentiate into cells of several lineages and contains cells from the microvasculature. The mechanisms that drive the ASCs into the osteoblast lineage are still not clear, but the process has been more extensively studied in bone marrow stromal cells. The purpose of this study was to investigate the osteogenic capacity of adipose derived SVF cells and evaluate bone formation following implantation of SVF cells into the bone defect of human phalanx. Methods: Case 1 a 43-year-old male was wounded while using a press machine. After first operation, segmental bone defects of the left 3rd and 4th middle phalanx occurred. At first we injected the SVF cells combined with demineralized bone matrix (DBM) to defected 4th middle phalangeal bone lesion. We used P (L/DL)LA [Poly (70L-lactide-co-30DL-lactide) Co Polymer P (L/DL)LA] as a scaffold. Next, we implanted the SVF cells combined with DBM to repair left 3rd middle phalangeal bone defect in sequence. Case 2 was a 25-year-old man with crushing hand injury. Three months after the previous surgery, we implanted the SVF cells combined with DBM to restore right 3rd middle phalangeal bone defect by syringe injection. Radiographic images were taken at follow-up hospital visits and evaluated radiographically by means of computerized analysis of digital images. Results: The phalangeal bone defect was treated with autologous SVF cells isolated and applied in a single operative procedure in combination with DBM. The SVF cells were supported in place with mechanical fixation with a resorbable macroporous sheets acting as a soft tissue barrier. The radiographic appearance of the defect revealed a restoration to average bone density and stable position of pharyngeal bone. Densitometric evaluations for digital X-ray revealed improved bone densities in two cases with pharyngeal bone defects, that is, 65.2% for 4th finger of the case 1, 60.5% for 3rd finger of the case 1 and 60.1% for the case 2. Conclusion: This study demonstrated that adipose derived stromal vascular fraction cells have osteogenic potential in two clinical case studies. Thus, these reports show that cells from the SVF cells have potential in many areas of clinical cell therapy and regenerative medicine, albeit a lot of work is yet to be done.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.29 no.6
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• pp.379-391
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• 2003

The present study was undertaken to evaluate bone regenerative capacity around titanium screw implants placed in irradiated rat's tibiae. At one week after single 15-Gy dose irradiation, miniaturized titanium screw implants were inserted into anterior aspect of the upper tibia of rats weighing 200-250g. Seventy rats were involved: 35 rats were control and 35 rats radiation group. The rats were killed at different intervals as 1, 2, 3, 4, 6, 8, 12 weeks after implantation for histologic observation, histomorphometric analysis and immunohistochemical study with fibronectin and CD34 antibody. 1. Histologically, various stages of bone maturation and ossification can be seen at 4 weeks and regenerated bone close to edges demonstrates more advanced calcification, and network of new bone are well formed at 12 weeks in non-irradiated group. In contrast, active bone formation with increased contact of newly formed bone to implant surface was noted at 4 weeks and a significant amount of new bone formation and bone-implant contact is oberved at 12 weeks in irradiated group. 2. Histomorphometrical analysis confirmed these histologic findings. A significant difference in implant-bone contact and bone density was measured between the control and radiation group. Mean MBD was 62.2% in control group and 27.5% in radiation group, mean MBIC was 86.6% in control group and 47.7% in radiation group, and mean TBIC was 87.3% in control group and 45.6% in radiation group at 12 weeks after implantation. 3. In immunohistochemical study with fibronectin and CD34, radiation reduced hematopoietic progenitor cells severely and disturbed differentiation of osteoblast in bone marrow. The results of this study revealed bone healing capacity around implant after radiation therapy was severely impaired and irradiation reduces the capacity for osseointegration of titanium implants. Many factors including radiation dose, period between radiation and implantation, bone quality, time elapse between first and second surgery, type of prosthetics and hyperbaric oxygen therapy must be considered carefully in postradiation implantation.

• Journal of Physiology & Pathology in Korean Medicine
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• v.34 no.1
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• pp.14-23
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• 2020

This study was conducted to evaluate the efficacy of Cheongawongagam on osteoporosis rat. A total of 35 rats were divided into seven groups; Normal control(SD-Nr), experimental control group(OVX-CTL), positive control group(OVX-17β-E2) and herb extracts group[Eucommia ulmoides(OVX-EU-E), Juglandis semen(OVX-JR-SE), Acanthopanax senticosus(OVX-AS-E) extract and Cheongawongagam extracts(OVX-JAEG-E)]. All control group, and herb extracts group were ovariectomized. After the 3 weeks recovery period, herb extract group were orally administered 200 mg / kg of the EU-E, JR-SE, AS-E and JAEG-E for 12 weeks. In the OVX-CTL, 17β-estradiol(E2) was administered subcutaneously on the back of the rats at a dose of 0.03 ug/sc. Their body weight, serum total cholesterol, triglyceride, aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), Leukotriene B4 (LTB4), calcium (Ca), estradiol, osteocalcin, and deoxypyridinoline (DPD) concentration were measured. Also, we investigated mRNA expression of inflammatory cytokine, MMP-2, MMP-9, and bone tissue. As a result, total cholesterol was significantly decreased in the OVX-AS-E and OVX-JAEG-E. ALP was significantly increased and osteocalcin, DPD was significantly decreased in OVX-JAEG-E. The expression of inflammatory mediators (TNF-α, IL-1β, LTB4, COX-2, NOS-2), inflammatory cytokines IL-1β and MMP-9 mRNA were significantly decreased in OVX-JAEG-E. Histologic examination of the femur showed that bone mineral density, and bone mass were increased and bone marrow were decreased in the OVX-JAEG-E group. The above experiment shows that cheongawongagam extract were effective in the prevention and treatment of osteoporosis.