• Title/Summary/Keyword: Rabbit. Histological Analysis

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BONE HEALING CAPACITY IN THE FRACTURE OF RABBIT MANDIBULAR BONE USING LOW-LEVEL LASER (가토 하악골 절단부의 저수준 레이저 처치가 골치유에 미치는 영향)

  • Bae, Yong-Hyeon;Han, Se-Jin;Kim, Kyung-Wook
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.35 no.2
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    • pp.120-124
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    • 2009
  • The concept of biostimulation of wounds by low-level laser therapy(LLLT) is attracting considerable attention. Although its effect on whole tissues has been studied quite extensively, the biological and cellular mechanisms underlying LLLT have not been clarified. In an experimental radius fracture in rabbits, Tang and Chai reported that LLLT enhanced the activity of red blood cells, macrophages, fibroblasts, chondrocytes, and osteoclasts within the fracture area. The purpose of the present study was to evaluate the effect of LLLT with a GaAlAs diode laser device on bone healing in rabbit mandibular fractures. We use 12 rabbits for this study. All rabbits were fractured mandible angle area using saw in anesthetic condition. In control group(n=6), none treatment was performed at fracture site. In experimental group(n=6), LLLT with a GaAlAs diode laser was radiated at fracture site daily for 7 days. All rabbits were sacrificed at 6 weeks later from performed fracture day. We studied the immunohistochemical staining of CD34 and Vimentin and the histochemical analysis for calcium and phosphorus content. The results were as follows. 1. In the histological and immunohistological staining, after 6week, fibroblasts, osteogenic cells and collgen fibers were observed more in experimental group than in control group. 2. In the histochemical analysis, the amount of calcium and phosphorus contents of the experimental group were more than the control group. From the results obtained, we suggest that the bone healing is stimulated by low-level laser irradiation in bone fractures.

Improvement of osteogenic potential of biphasic calcium phosphate bone substitute coated with synthetic cell binding peptide sequences

  • Choi, Hyunmin;Park, Nho-Jae;Jamiyandorj, Otgonbold;Hong, Min-Ho;Oh, Seunghan;Park, Young-Bum;Kim, Sungtae
    • Journal of Periodontal and Implant Science
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    • v.42 no.5
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    • pp.166-172
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    • 2012
  • Purpose: The aim of this study was to evaluate the improvement of osteogenic potential of biphasic calcium phosphate (BCP) bone substitute coated with synthetic cell-binding peptide sequences in a standardized rabbit sinus model. Methods: Standardized 6-mm diameter defects were created bilaterally on the maxillary sinus of ten male New Zealand white rabbits, receiving BCP bone substitute coated with synthetic cell binding peptide sequences on one side (experimental group) and BCP bone substitute without coating (control group) on the other side. Histologic and histomorphometric analysis of bone formation was carried out after a healing period of 4 or 8 weeks. Results: Histological analysis revealed signs of new bone formation in both experimental groups (4- and 8-week healing groups) with a statistically significant increase in bone formation in the 4-week healing group compared to the control group. However, no statistically significant difference in bone formation was found between the 8-week healing group and the control group. Conclusions: This study found that BCP bone substitute coated with synthetic cell-binding peptide sequences enhanced osteoinductive potential in a standardized rabbit sinus model and its effectiveness was greater in the 4-week healing group than in the 8-week healing group.

Histological Comparative Study of Rabbit Maxillary Sinus Augmentation with Bio-Oss and β-TCP (Bio-Oss와 β-TCP를 이용한 토끼 상악동 거상술 후의 조직학적 비교 연구)

  • Moon, Yong-Suk
    • Journal of Life Science
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    • v.28 no.10
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    • pp.1220-1232
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    • 2018
  • The purpose of this animal study was to evaluate, by histological analysis, bone regeneration in rabbit maxillary sinuses with an anorganic bovine graft (Bio-Oss) and a ${\beta}-tricalcium$ phosphate (${\beta}-TCP$) grafting. Bilateral sinus augmentation procedures were performed in 12 adult male rabbits. Rectangular replaceable bony windows were made with a piezoelectric thin saw insert. In the Bio-Oss group, Bio-Oss was grafted and in the ${\beta}-TCP$ group, ${\beta}-TCP$ was grafted and covered by replaceable bony windows. The animals were sacrificed at 2, 4, and 8 weeks after the surgical procedure. The augmented sinuses were evaluated by histomorphometric analysis using hematoxylin-eosin, Masson trichrome, and tartrate-resistant acid phosphatase stains and also by immunohistochemical analysis of proliferating cell nuclear antigen (PCNA), type I collagen, and osteocalcin content. Histologically, new bone formation was found on the surface of Bio-Oss and ${\beta}-TCP$ particles from 2 weeks and continued to 8 weeks. Significant higher new bone formation was revealed in the ${\beta}-TCP$ group than in the Bio-Oss group at 8 weeks. The amount of graft materials was significantly decreased in the ${\beta}-TCP$ group and the number of osteoclasts was significantly increased in the ${\beta}-TCP$ group from 4 to 8 weeks. Immunoreactivity to PCNA was reduced at 8 weeks. The expression of type I collagen was significantly increased in the ${\beta}-TCP$ group at 2 weeks, but was significantly increased in the Bio-Oss group at 8 weeks. Immunoreactivity to osteocalcin was increased from 2 to 8 weeks. These histological results can help in the selection of graft materials for implants. Both Bio-Oss and ${\beta}-TCP$ are proven graft materials, however, these results indicate that ${\beta}-TCP$ showed better bone regeneration results in rabbit maxillary sinus augmentation.

Comparison of the bone healing capacity of autogenous bone, demineralized freeze dried bone allograft, and collagen sponge in repairing rabbit cranial defects

  • Hur, Jung-Woo;Yoon, Suk-Ja;Ryu, Sun-Youl
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.38 no.4
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    • pp.221-230
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    • 2012
  • Objectives: This study sought to evaluate the efficacy of collagen graft materials, as compared to other graft materials, for use in healing calvarial defects in rabbits. Materials and Methods: Ten mm diameter calvarial defects were made in ten rabbits. The rabbits were then divided into 4 groups: control, autogenous bone graft, SureOss graft, and Teruplug graft. Bone regeneration was evaluated using histological and radiographic methods. Results: Based on visual examination, no distinct healing profile was observed. At 4 weeks after treatment, histological analysis showed there was no bone regeneration in the control group; however, at 8 weeks after treatment, new bone formation was observed around the margin of the defective sites. In the autogenous bone graft group, new bone formation was observed at 4 weeks after treatment and mature bone was detected around the grafted bone after 8 weeks. In the SureOss graft group, at 4 weeks after treatment, acute inflammatory and multinuclear cells were noted around the grafted materials; at 8 weeks after treatment, a decrease in graft materials coupled with new bone formation were observed at the defective sites. In the Teruplug graft group, new bone formation was detected surrounding the bone margin and without signs of inflammation. There were statistically significant differences observed between the graft and control group in terms of bone density as evidenced by radiographic analysis using computed tomography (P<0.05), particularly for the autogenous bone graft group (P<0.001). Conclusion: These results suggested that autogenous bone, SureOss and Teruplug have the ability to induce bone regeneration as compared to an untreated control group. The osteogenic potential of Teruplug was observed to be lower than that of autogenous bone, but similar to that of SureOss.

Comparative analysis of carrier systems for delivering bone morphogenetic proteins

  • Jung, Im-Hee;Lim, Hyun-Chang;Lee, Eun-Ung;Lee, Jung-Seok;Jung, Ui-Won;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.45 no.4
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    • pp.136-144
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    • 2015
  • Purpose: The objective of this study was to comparatively assess the bone regenerative capacity of absorbable collagen sponge (ACS), biphasic calcium phosphate block (BCP) and collagenated biphasic calcium phosphate (CBCP) loaded with a low dose of recombinant human bone morphogenetic protein-2 (rhBMP-2). Methods: The CBCP was characterized by X-ray diffraction and scanning electron microscopy. In rabbit calvaria, four circular 8-mm-diameter defects were created and assigned to one of four groups: (1) blood-filled group (control), (2) rhBMP-2-soaked absorbable collagen sponge (0.05 mg/mL, 0.1 mL; CS group), (3) rhBMP-2-loaded BCP (BCP group), or (4) rhBMP-2-loaded CBCP (CBCP group). The animals were sacrificed either 2 weeks or 8 weeks postoperatively. Histological and histomorphometric analyses were performed. Results: The CBCP showed web-like collagen fibrils on and between particles. Greater dimensional stability was observed in the BCP and CBCP groups than in the control and the CS groups at 2 and 8 weeks. The new bone formation was significantly greater in the BCP and CBCP groups than in the control and CS groups at 2 weeks, but did not significantly differ among the four groups at 8 week. The CBCP group exhibited more new bone formation in the intergranular space and in the center of the defect compared to the BCP group at 2 weeks, but a similar histologic appearance was observed in both groups at 8 weeks. Conclusions: The dose of rhBMP-2 in the present study enhanced bone regeneration in the early healing period when loaded on BCP and CBCP in rabbit calvarial defects.

The Effect of a Hydroxyapatite and 4-hexylresorcinol Combination Graft on Bone Regeneration in the Rabbit Calvarial Defect Model (가토의 두정골 결손부 모델에서 수산화인회석에 4-hexylresorcinol을 혼합하여 만든 인공합성골이식재의 골형성효과에 대한 연구)

  • Kim, Min Keun;Park, Yong Tae;Kim, Seong-Gon;Park, Young-Wook;Lee, Suk-Keun;Choi, Weon-Sik
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.34 no.6
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    • pp.377-383
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    • 2012
  • Purpose: The aim of this study was to evaluate the effect of 4-hexylresorcinol and hydroxyapatite combination graft on bone regeneration in the rabbit calvarial defect model. Methods: Ten New Zealand white rabbits were used for this study. Bilateral round shaped defects (diameter: 8.0 mm) were created on the parietal bone. 4-hexylresorcinol and hydroxyapatite combination graft material was grafted into the right parietal bone defect area (experimental). The left bone defect area was not filled with anything (control). The animals were sacrificed at 4 weeks and 8 weeks after grafting. A micro-computerized tomography of each specimen was taken, and the specimens were stained for histological analysis. Results: The average value of bone mineral density (BMD) and Bone volume (BV) was higher in the experimental group than in the control group at 4 weeks and 8 weeks after surgery. However, the difference was not statistically significant (P>0.05) at 8 weeks after grafting. The BMD and BV in the experimental group at 4 weeks after surgery was significantly higher than those in the control group (P<0.05). Conclusion: 4-hexylresorcinol and hydroxyapatite combination graft material showed higher initial bone formation than the control, however, there was no difference at 8weeks after operation.

Effects of Ischemic Preconditioning of Different Intraoperative Ischemic Times of Vascularized Bone Graft Rabbit Models

  • Halim, Ahmad Sukari;Wan Ahmad Kamal, Wan Syazli Rodzaian;Noor, Norizal Mohd;Abdullah, Shafie
    • Archives of Plastic Surgery
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    • v.40 no.6
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    • pp.687-696
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    • 2013
  • Background Ischemic preconditioning has been shown to improve the outcomes of hypoxic tolerance of the heart, brain, lung, liver, jejunum, skin, and muscle tissues. However, to date, no report of ischemic preconditioning on vascularized bone grafts has been published. Methods Sixteen rabbits were divided into four groups with ischemic times of 2, 6, 14, and 18 hours. Half of the rabbits in each group underwent ischemic preconditioning. The osteomyocutaneous flaps consisted of the tibia bone, from which the overlying muscle and skin were raised. The technique of ischemic preconditioning involved applying a vascular clamp to the pedicle for 3 cycles of 10 minutes each. The rabbits then underwent serial plain radiography and computed tomography imaging on the first, second, fourth, and sixth postoperative weeks. Following this, all of the rabbits were sacrificed and histological examinations were performed. Results The results showed that for clinical analysis of the skin flaps and bone grafts, the preconditioned groups showed better survivability. In the plain radiographs, except for two non-preconditioned rabbits with intraoperative ischemic times of 6 hours, all began to show early callus formation at the fourth week. The computed tomography findings showed more callus formation in the preconditioned groups for all of the ischemic times except for the 18- hour group. The histological findings correlated with the radiological findings. There was no statistical significance in the difference between the two groups. Conclusions In conclusion, ischemic preconditioning improved the survivability of skin flaps and increased callus formation during the healing process of vascularized bone grafts.

Evaluation of bone substitutes for treatment of peri-implant bone defects: biomechanical, histological, and immunohistochemical analyses in the rabbit tibia

  • dos Santos, Pamela Leticia;de Molon, Rafael Scaf;Queiroz, Thallita Pereira;Okamoto, Roberta;de Souza Faloni, Ana Paula;Gulinelli, Jessica Lemos;Luvizuto, Eloa Rodrigues;Garcia, Idelmo Rangel Junior
    • Journal of Periodontal and Implant Science
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    • v.46 no.3
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    • pp.176-196
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    • 2016
  • Purpose: We sought to evaluate the effectiveness of bone substitutes in circumferential periimplant defects created in the rabbit tibia. Methods: Thirty rabbits received 45 implants in their left and right tibia. A circumferential bone defect (6.1 mm in diameter/4 mm depth) was created in each rabbit tibia using a trephine bur. A dental implant ($4.1mm{\times}8.5mm$) was installed after the creation of the defect, providing a 2-mm gap. The bone defect gaps between the implant and the bone were randomly filled according to the following groups: blood clot (CO), particulate Bio-Oss$^{(R)}$ (BI), and Bio-Oss$^{(R)}$ Collagen (BC). Ten animals were euthanized after periods of 15, 30, and 60 days. Biomechanical analysis by means of the removal torque of the implants, as well as histologic and immunohistochemical analyses for protein expression of osteocalcin (OC), Runx2, OPG, RANKL, and TRAP were evaluated. Results: For biomechanics, BC showed a better biological response ($61.00{\pm}15.28Ncm$) than CO ($31.60{\pm}14.38Ncm$) at 30 days. Immunohistochemical analysis showed significantly different OC expression in CO and BC at 15 days, and also between the CO and BI groups, and between the CO and BC groups at 60 days. After 15 days, Runx2 expression was significantly different in the BI group compared to the CO and BC groups. RANKL expression was significantly different in the BI and CO groups and between the BI and BC groups at 15 days, and also between the BI and CO groups at 60 days. OPG expression was significantly higher at 60 days postoperatively in the BI group than the CO group. Conclusions: Collectively, our data indicate that, compared to CO and BI, BC offered better bone healing, which was characterized by greater RUNX2, OC, and OPG immunolabeling, and required greater reversal torque for implant removal. Indeed, along with BI, BC presents promising biomechanical and biological properties supporting its possible use in osteoconductive grafts for filling peri-implant gaps.

EPMA analysis of bone formation around RBM surface implant (RBM surface 임플란트의 골형성에 대한 EPMA 분석)

  • Jung, Hwan-Jong;Yim, Sung-Bin;Chung, Chin-Hyung;Hong, Ki-Seok
    • Journal of Periodontal and Implant Science
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    • v.38 no.3
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    • pp.503-510
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    • 2008
  • Purpose: To evaluate bone formation on the implant surface between machined fixture and RBM surface fixture through analyzing of distribution and concentration of Ca and P by EPMA. Material and Methods: Fixtures had divided in 2 group: Machined(Group I), RBM (Group II). Total 4 fixtures were implanted on rabbit which sacrificed on 2 week and 4 week for the histological specimens. By these specimens EPMA value were measured, compared and analysed by each group to figure out the evidence of clinical use of RBM implant. Result: After 2 weeks and 4 weeks, it was analyzed that bone formation area, distribution and concentration of Ca and P by EPMA. In distribution and concentration of P Group II was higher than Group I, but there were no statistical significances. In new bone formation area, Group II was more higher than Group I with statistically significances. Both of group, after 4 weeks area is little bit higher than after 2 weeks area but there is no statistically significances. Conclusion: RBM implant was better than machined implant on the early bone formation.

THE EXPRESSION OF TGF-$\beta$1, IGF-I, BFGF IN DISTRACTION OSTEOGENESIS ACCORDING TO DIFFERENT DISTRACTION RATES IN RABBIT'S MANDIBLE (가토 하악골에서 신연 골형성술시 신연속도에 따른 TGF-$\beta$1, IGF-I, bFGF의 발현)

  • Shin, Sun-A;Jee, Yu-Jin;Song, Hyun-Chul
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.27 no.3
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    • pp.205-217
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    • 2005
  • Distraction osteogenesis is a technique of lengthening bone including soft tissue by gradual separation of surgically divided bone surfaces. Although the biomechanical, histological, and ultrastructural changes associated with distraction osteogenesis have been widely described, the molecular mechanisms governing the formation of new bone in distracted bone segments remain largely unclear. However, such information has significant clinical implications because it may enable targeted therapeutic manipulations designed to accelerate osseous regeneration. The purpose of this study was to evaluate the expression of TGF-$\beta$1, IGF-I and bFGF in distraction osteogenesis according to different distraction rates in a rabbit's mandible. When twenty-four adult rabbits underwent open osteotomy between the premolar and mental foramen, an external bilateral distraction device was applied. Latency was allowed for five days before distraction. Three different distraction rates were 0.7 mm/day (A, n=8), 1.4 mm/day (B, n=8) and 2.4 mm/day (C, n=8). The distraction device was activated with the same distraction rhythms of twice a day until 4.9 mm (A & B group) and 8.4 mm (C group) length gains was achieved. The animals were sacrificed at postoperative 3, 7, 14 and 28 days. The bony specimens were stained with H&E for histologic examination, and RT-PCR analysis was done for the identification of the expression of TGF-$\beta$1, IGF-I and bFGF. The results obtained from this study were as follows : The 0.7 mm/day and 1.4 mm/day distraction rate groups were shown to improve regenerative bone formation on radiographic and histologic examination. Also, TGF-$\beta$1, IGF-I and bFGF expression increased in the 0.7 mm/day and 1.4 mm/day distraction rate groups. But the 2.4 mm/day distraction rate group specimen was different with adjacent normal bone and hardly expressed of growth factors. These findings suggest that improved new bone formation in the 0.7 mm/day and 1.4 mm/day distraction rates is associated with enhanced expression of TGF-$\beta$1, IGF-I and bFGF by mechanical tension stress. Additionally, the 0.7 mm/day and 1.4 mm/day distraction rate groups were significantly different from the 2.4 mm/day distraction rate group in the expression of growth factors. According to the above results, it seems possible to apply a distraction rate of up to 1.4 mm/day a day in rabbit's mandible. And further studies are needed to evaluate growth factors of TGF-$\beta$1 and IGF-I, which are excellent in expression.