Purpose: Adipose tissue is located beneath the skin, around internal organs, and in the bone marrow in humans. Its main role is to store energy in the form of fat, although it also cushions and insulates the body. Adipose tissue also has the ability to dynamically expand and shrink throughout the life of an adult. Recently, it has been shown that adipose tissue contains a population of adult multipotent mesenchymal stem cells and endothelial progenitor cells that, in cell culture conditions, have extensive proliferative capacity and are able to differentiate into several lineages, including, osteogenic, chondrogenic, endothelial cells, and myogenic lineages. Materials and Methods: This study focused on endothelial cell culture from the adipose tissue. Adipose tissues were harvested from buccal fat pad during bilateral sagittal split ramus osteotomy for surgical correction of mandibular prognathism. The tissues were treated with 0.075% type I collagenase. The samples were neutralized with DMEM/and centrifuged for 10 min at 2,400 rpm. The pellet was treated with 3 volume of RBC lysis buffer and filtered through a 100 ${\mu}m$ nylon cell strainer. The filtered cells were centrifuged for 10 min at 2,400 rpm. The cells were further cultured in the endothelial cell culture medium (EGM-2, Cambrex, Walkersville, Md., USA) supplemented with 10% fetal bovine serum, human EGF, human VEGF, human insulin-like growth factor-1, human FGF-$\beta$, heparin, ascorbic acid and hydrocortisone at a density of $1{\times}10^5$ cells/well in a 24-well plate. Low positivity of endothelial cell markers, such as CD31 and CD146, was observed during early passage of cells. Results: Increase of CD146 positivity was observed in passage 5 to 7 adipose tissue-derived cells. However, CD44, representative mesenchymal stem cell marker, was also strongly expressed. CD146 sorted adipose tissue-derived cells was cultured using immuno-magnetic beads. Magnetic labeling with 100 ${\mu}l$ microbeads per 108 cells was performed for 30 minutes at $4^{\circ}C$ a using CD146 direct cell isolation kit. Magnetic separation was carried out and a separator under a biological hood. Aliquous of CD146+ sorted cells were evaluated for purity by flow cytometry. Sorted cells were 96.04% positivity for CD146. And then tube formation was examined. These CD146 sorted adipose tissue-derived cells formed tube-like structures on Matrigel. Conclusion: These results suggest that adipose tissue-derived cells are endothelial cells. With the fabrication of the vascularized scaffold construct, novel approaches could be developed to enhance the engineered scaffold by the addition of adipose tissue-derived endothelial cells and periosteal-derived osteoblastic cells to promote bone growth.
Purpose: To prolong the degradation time of collagen membranes, various cross-linking techniques have been developed. For cross-linking, chemicals such as formaldehyde and glutaraldehyde are added to collagen membranes, but these chemicals could adversely affect surrounding tissues. The aim of this study is to evaluate the ability of porous non-chemical cross-linking porcine-derived collagen nanofibrous membrane to enhance bone and associated tissue regeneration in one-wall intrabony defects in beagle dogs. Methods: The second and third mandibular premolars and the first molars of 2 adult beagles were extracted bilaterally and the extraction sites were allowed to heal for 10 weeks. One-wall intrabony defects were prepared bilaterally on the mesial and distal side of the fourth mandibular premolars. Among eight defects, four defects were not covered with membrane as controls and the other four defects were covered with membrane as the experimental group. The animals were sacrificed 10 weeks after surgery. Results: Wound healing was generally uneventful. For all parameters evaluating bone regeneration, the experimental group showed significantly superior results compared to the control. In new bone height (NBh), the experimental group exhibited a greater mean value than the control ($3.04{\pm}0.23\;mm/1.57{\pm}0.59$, P=0.003). Also, in new bone area (NBa) and new bone volume (NBv), the experimental group showed superior results compared to the control (NBa, $34.48{\pm}10.21%$ vs. $5.09{\pm}5.76%$, P=0.014; and NBv, $28.04{\pm}12.96$ vs. $1.55{\pm}0.57$, P=0.041). On the other hand, for parameters evaluating periodontal tissue regeneration, including junctional epithelium migration and new cementum height, there were no statistically significant differences between two groups. Conclusions: Within the limitations of this study, this collagen membrane enhanced bone regeneration at one-wall intrabony defects. On the other hand, no influence of this membrane on periodontal tissue regeneration could be ascertained in this study.
Kim, Tae-Sung;Lee, Dong-Keun;Lee, Byung-Do;Jung, Sun-Kwan
Maxillofacial Plastic and Reconstructive Surgery
/
v.23
no.2
/
pp.107-114
/
2001
Objectives : To compare jaw bone density of young adults (control group) and post-menopausal women(experimental group) in periapical and panoramic film. Materials and Methods : The bone mineral density values of lumbar and femur were measured by dual-energy X-ray absorptiometry(DEXA) and T scores of lumbar were obtained. T scores were classified into 3 group (T<-2.5, $-2.5{\leqq}T<-1$, $-1{\leqq}T$). Radiographic densities of alveolar bones were measured from interdental bones of premolar, molar areas in the maxilla and mandible and expressed into copper step wedge thickness by Scion $Image^{(R)}$ program. We considered these values of step wedge thickness as bone density of alveolar bone. Panorama mandibular index(PMI) was calculated by the method that the height of the inferior cortex of the mandible was divided by the height from the lower border of the mandible to the superior edge of the mental foramen. Bone density of alveolar bone and PMI were analysed statistically. Results : There were significant differences in bone mineral density of lumbar and femoral neck between control and experimental groups. There were also significant differences in bone density of premolar and molar area of jaw between control and experimental groups by MANOVA test. When considered lumbar T variables, there was only difference in interdental bone density of maxillary molar area between control and experimental group, but there was interaction. Interdental bone density of experimental group was appeared higher in $-1{\leqq}T$ group and lower in T<-2.5 group than control group. There was significant difference in PMI between control and experimental groups, but there was also inter action, thus, PMI of experimental group was appeared higher in $-1{\leqq}T$ group and lower in T<-2.5 group than control group. Conclusion : There were significant differences of alveolar density and cortical bone thickness between young men and post-menopausal women in periapical and panoramic film. These differences were dependent on lumbar T.
Background: The aim of this study was to evaluate the effect of autogenous tooth bone as a graft material for regeneration of bone in vertical bony defects of the minipigs. Material and Methods: Six minipigs were used in this study. Four molars were extracted in the right mandibular dentition and sent to the Korea Tooth Bank for fabrication of autogenous tooth bone. Ten days later, each extraction site was implanted with MS Implant Narrow Ridge $3.0{\times}10mm$ fixture (Osstem, Seoul, Korea) after standardized 2mm-sized artificial vertical bony defect formation. Pineappleshaped Root-On type autogenous tooth bones were applied to the vertical defects around the neck area of the posterior three fixtures and the fore-most one was not applied with autogenous bone as a control group. Each minipig was sacrificed at 4, 8, 12 weeks after fixture installation and examined radiologically and histologically. Histological evaluation was done under light microscope with Villanueva osteochrome bone staining with semi-quantitative histomorphometric study. Percentage of new bone over total area (NBF) and bone to implant contact (BIC) ratio were evaluated using digital software for area calculation. Result: NBF were $48.15{\pm}18.02%$, $45.50{\pm}28.37%$, and $77.13{\pm}15.30%$ in 4, 8, and 12 weeks, respectively for experimental groups. The control group showed $37.00{\pm}11.53%$, $32.25{\pm}26.99%$, and $1.33{\pm}2.31%$ in 4,8,12 weeks, respectively. BIC ratio were $53.08{\pm}19.82%$, $45.00{\pm}28.37%$, and $75.13{\pm}16.55%$ in 4,8,12 weeks, respectively. Those for the control groups were $38.33{\pm}6.43%$, $33.50{\pm}29.51%$, and $1.33{\pm}2.31%$ in 4, 8, 12 weeks, respectively. Conclusion: Autogenous tooth bone showed higher score than control group in NBF and BIC in all the data encompassing 4,8,12 weeks specimens, but statistically significant only 12 weeks data in both NBF and BIC.
The author has arrived at the following result after having carried out multilateral study based on a total of 282 maxillofacial fracture patients who have receive treatment at the Euijeongbu general hospital and Shinchun general hospital in the northern district of Kyunggido from march 1988 to august 1990. 1. Sex distribution of Mx. facial fx. patient was higher in male by 4.6:1 and was predominant in the 3rd decade with 40.4% followed in decreasing order by the 2nd decade and the 4th. 2. A majority were in the Mn. with 40.2% followed in decreasing order by zygoma. nasal bone and maxilla. 3. For the sex distribution according to anatomy, make to female ratio was 6.2:1 in the mandible, followed in decreasing order by zygoma, and nasal bone with predominance in male. 4. Car accident with 42.8% was the most common cause of fx. followed in decreasing order by violence, workmen's accident, and fall down. 5. The involvement of other trauma areas are head. 79.0%, abdomen-thorax, and the extremities in decreasing order. 6. In the mandibular fx. a majority were in the symphysis with 73.9% followed in decreasing order by angle, Condyle, and body. 7. Maxillary fx. of the type LeFort II was estimated to be 41.2% 8. Fracture in the zygoma including zygomatic arch was estimated to be 72.5%
Kim, Uk-Kyu;Shin, Sang-Hun;Chung, In-Kyo;Kim, Cheol-Hun;Huo, Jun;Yun, Il
Maxillofacial Plastic and Reconstructive Surgery
/
v.27
no.5
/
pp.403-414
/
2005
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.
The fibrous layer of mandibular condyle of the neonatal, 1-, 7-, 14-, 27-, 55-days and 1 year old rats were examined in the electron microscope with particular attention to the ultrastructure and diameter of collagen fibrils. In the 1-day rats, most of the cells of the fibrous layer were undifferentiated mesenchymal cells and fibroblasts with rough a little developed rough endoplasmic reticulum(RER) and golgi apparatus(GA). In 7-, 17 and 27-days old rats, most of the fibroblast showed well developed GA and RER with widely distended cisternae containing granular materials. In many of these cells contained intracytoplasmic filaments among the cytoplamic organelle. In 55-day and 1-year old rats, three types of cells were observed, ie, cells containing well developed cytoplasmic organelle presumed to be involved in the collagen fibril synthesis, cells showing well developed lysosomes, golgi apparatus, mitochondria and short cytoplasmic process presumed to be involved in the active resorption of the injured collagen fibrils or cellular debris, cells containing many intracytoplasmic filaments and a little organelle presumed to be cells of inactive state. The average diameters of collagen fibrils were similar in 1- and 7-day old rats as $38.48{\pm}3.81nm$, $38.06{\pm}3.86nm$. That was thickest in 14 days old rats as $50.21{\pm}3.93nm$ among experimental groups. They were gradually thinner in 27-, 55-day rats as $40.05{\pm}2.52nm$, $43.63{\pm}1.20nm$ and thinnest in 1-year old rats as $37.38{\pm}2.17nm$. The distribution pattern of diameters of collagen fibrils were unimordal with peak of 30-60nm in rats from 1-day to 17-day old. With aging from 27-day to 1 year old rats, collagen fibril diameters showed wide distribution pattern and percentage of thin collagen fibrils increased. These results may show the functional adaptation of fibrous layer of mandibular condyle to the increased mechanical forces with aging.
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