Many researches have been reported that collagen as cellular stroma, matrix of grafting materials, mediator of agents for the purpose of promoting healing process invivo, but the responses in vivo were seen various. The goal of this experiment is to assess the effect of collagen on bony healing, through histological evaluation of implanted collagen on the calvarial defect in rats. 2-month-old Sprague-Dawley, 24 rats were used and 12 rats assigned to each group of control and test. Defect of 5mm in diameter was made on the calvarial bone with trephine bur. Following thorough saline rinse, defect of control group was left in empty and that of experimental group was filled with fibrillar collagen($COLLATAPE^{(R)}$, COLLA-TEC. INC. U.S.A.) soaked in saline. 3 rats in each group were sacrificed at 3, 7, 14, 21 days after operation respectively, and the tissue blocks were prepared for light microscope with H-E for evaluation of overall healing, with TRAP(tartrate resistant acid phosphatase) for evaluation of osteoclastic activity and with immunohistochemical staining for macrophages. The results were as follows : 1. In the control group, inflammatory responses were disappeared at day 14, but, in the experimental group inflammatory infiltrates were reduced at day 21. Thus, the experimental group showed more severe soft tissue inflammation than control group. 2. Both control and experimental group showed slight appositional growth at day 7 and gradual bony growth to 21th day. But, complete bony healing of the defect was not shown. There was no significant difference in bony healing between control and experimental group 3. Specific response of macrophages for implanted collagen was observed at day 14 in the experimental group. In conclusion, although fibrillar collagen caused inflammation of soft tissue during initial healing period, inflammatory responses by fibrillar collagen didn't inhibit bony regeneration and implanted collagen was biodegradaded by macrophages. Thus, we expect that fibrillar collagen can be used for useful mediator of graft materials or growth factors.
The purpose of this study was to evaluate the efficacy of adding autogenous bone to the toothash-plaster mixture in the healing process of bone. Full-thickness round osseous defects with the diameter of 20mm were made at the calvarial bone of adult dogs (n=19) bilaterally, which were thought to be critical size defect. The right defects were repaired with the toothash-plaster mixture plus autogenous bone (compressed volume 0.3cc) and the left defects with only toothash-plaster mixture. At 2-, 4-, 8-, 12- and 20- week after implantation, dogs were sacrificed and evaluated the osseous healing of bony defects clinically, radiographically, and microscopically. The results were as follows; 1. At the clinical observation, the wound healed very well without any problem except severe swelling in the early period after operation. Slight depression was recognized at the both sides when the portions of cranial defect were palpated. 2. There were statistically significant differences between toothash-plaster mixture groups and autogenous bone added groups at the same period, and among the groups in the bone density of the digital radiograms (P<0.001). There was a tendency that bone density was increasing with time. 3. In light microscopic examination, new bone formation was more active in the autogenous bone added groups than toothash-plaster mixture groups at the early period after implantation but there is little difference at 20-week after implantation. 4. In fluorescent microscopic examination, the fluorescent band could be observed at the area of active bone formation and the band was more distinct in the autogenous bone added groups then toothash-plaster mixture groups. 5. In transmitted electron microscopic examination, organelles such as rER, Golgi complex and secretory granule and osteoblast were observed. In summary higher volume ratio of autogenous bone is needed to improve the bone healing in that there is little difference between toothash-plaster mixture group and autogenous bone added group at the 20-week after implantation in spite of new bone formation was more active in the autogenous bone added groups than toothash-plaster mixture groups at the early period after operation.
Purpose: Recombining bone morphogenetic protein (BMP) is usually acquiredfrom high level animals. Though this method is effective, its high cost limits its use. The purpose of this study was to evaluate the effect of bone morphogenetic protein-2 with protein transduction domain (BMP-2/PTD;TATBMP-2) on bone regeneration. Rat calvarial defect model and osteoblastic differentiation model using MC3T3 cell were used for the purpose of the study. Materials and Methods: MC3T3 cells were cultured until they reached a confluence stage. The cells were treated with 0, 0.1, 1, 10, 100, 500 ng/ml of BMP-2/PTD for 21 days and at the end of the treatment, osteoblastic differentiation was evaluated usingvon Kossa staining. An 8mm, calvarial, critical-size osteotomy defect was created in each of 48 male Spraque-Dawley rats (weight $250{\sim}300\;g$). Three groups of 16 animals each received either BMP-2/PTD (0.05mg/ml) in a collagen carrier, collagen only, or negative surgical control. And each group was divided into 2 and 8 weeks healing intervals. The groups were evaluated by histologic analysis(8 animals/group/healing intervals) Result: In osteoblastic differentiation evaluation test, a stimulatory effect of BMP-2/PTD was observed in 10ng/ml of BMP-2/PTD with no observation of dose-dependent manner. The BMP-2/PTD group showed enhanced local bone formation in the rat calvarial defect at 2 weeks. New bone was observed at the defect margin and central area of the defect. However, new bone formation was observed only in 50% of animals used for 2weeks. In addition, there was no new bone formation observed at 8 weeks. Conclusion: The results of the present study indicated that BMP-2/PTD(TATBMP-2) have an positive effect on the bone formation in vitro and in vivo. However, further study should be conducted for the reproducibility of the outcomes.
Montevecchi, Marco;Parrilli, Annapaola;Fini, Milena;Gatto, Maria Rosaria;Muttini, Aurelio;Checchi, Luigi
Journal of Periodontal and Implant Science
/
v.46
no.5
/
pp.303-319
/
2016
Purpose: The purpose of this animal study was to perform a 3-dimensional micro-computed tomography (micro-CT) analysis in order to investigate the influence of root surface distance to the alveolar bone and the periodontal ligament on periodontal wound healing after a guided tissue regeneration (GTR) procedure. Methods: Three adult Sus scrofa domesticus specimens were used. The study sample included 6 teeth, corresponding to 2 third mandibular incisors from each animal. After coronectomy, a circumferential bone defect was created in each tooth by means of calibrated piezoelectric inserts. The experimental defects had depths of 3 mm, 5 mm, 7 mm, 9 mm, and 11 mm, with a constant width of 2 mm. One tooth with no defect was used as a control. The defects were covered with a bioresorbable membrane and protected with a flap. After 6 months, the animals were euthanised and tissue blocks were harvested and preserved for micro-CT analysis. Results: New alveolar bone was consistently present in all experimental defects. Signs of root resorption were observed in all samples, with the extent of resorption directly correlated to the vertical extent of the defect; the medial third of the root was the most commonly affected area. Signs of ankylosis were recorded in the defects that were 3 mm and 7 mm in depth. Density and other indicators of bone quality decreased with increasing defect depth. Conclusions: After a GTR procedure, the periodontal ligament and the alveolar bone appeared to compete in periodontal wound healing. Moreover, the observed decrease in bone quality indicators suggests that intrabony defects beyond a critical size cannot be regenerated. This finding may be relevant for the clinical application of periodontal regeneration, since it implies that GTR has a dimensional limit.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
/
v.30
no.4
/
pp.282-291
/
2004
Pure-phase beta-tricalcium phosphate(${\beta}$-TCP) proved to be a bone regeneration material, providing the patient with vital bone at the defect site in a reasonable time, making a second surgical procedure for bone harvesting unnecessary. This study compares bone healing and BMP 2/4 expression in cranial defects in rabbits grafted with autogenous bone and ${\beta}$-TCP. Thirty New Zealand White rabbits was divided into 3 group of 10 animals each. Bilateral calvarial defects were made in the parietal bones of each animal. ${\beta}$-TCP placed in one defect and the other defects was filled with autogenous bone. The animal were sacrificed at 4, 8 and 12 weeks. Immunohistochemical analysis was used to investigate the expression of BMP 2/4. 1. The new bone formation around autogenous bone from 4 weeks and ${\beta}$-TCP from 8 weeks. 2. In autogenous bone graft, BMP 2/4 expression was decreased from 4 to 12 weeks. 3. In ${\beta}$-TCP graft, BMP 4 expression was increased from 8 to 12 weeks. But, BMP 2 was observed from 12 weeks. This study showed that bone healing, regeneration and, BMP 2/4 expression are delayed in grafted ${\beta}$-TCP than autogenous bone.
An, Min Kuk;Kim, Hyun Ju;Choi, Jin Uk;Kim, Kyoung-Hwa;Lee, Yong-Moo;Rhyu, In-Chul;Seol, Yang-Jo
Journal of Periodontal and Implant Science
/
v.52
no.5
/
pp.422-434
/
2022
Purpose: The purpose of this study was to evaluate and compare the healing patterns of 2-mm and 4-mm proximal infrabony defects adjacent to dental implants in canine mandibles. Methods: Four male beagles were used. Two groups were created: a 2-mm group (n=4) and a 4-mm group (n=4) depending on the horizontal dimension of proximal infrabony defects adjacent to implants. Bone healing patterns between the 2 groups were evaluated and compared at 8 and 16 weeks using radiographic, histological, histomorphometric, and fluorescent labelling analyses. Results: According to microcomputed tomography, the median bone volume fraction, bone mineral density, and the percentage of radiographic distance from the defect bottom to the most coronal bone-to-implant contact (radio-mcBIC) were 32.9%, 0.6 g/cm3, and 73.7% (8 weeks) and 45.7%, 0.7 g/cm3, and 76.0% (16 weeks) in the 2-mm group and 57.7%, 0.8 g/cm3, and 75.7% (8 weeks) and 50.9%, 0.8 g/cm3, and 74.7% (16 weeks) in the 4-mm group, respectively. According to histomorphometry, the median bone area fraction, mcBIC and the percentage of BIC amounted to 36.7%, 3.4 mm, and 58.4% (8 weeks) and 49.2%, 3.4 mm, and 70.2% (16 weeks) in the 2-mm group and 50.0%, 3.0 mm, and 64.8% (8 weeks) and 55.7%, 3.0 mm, and 69.6% (16 weeks) in the 4-mm group, respectively. No statistically significant differences were found between the groups for any variables (P>0.05). Conclusions: The proximal defects that measured 2 mm and 4 mm showed similar healing patterns at 8 and 16 weeks, and the top of bone formation in the defects was substantially limited to a maximum of 1.6 mm below the implant shoulder in both groups.
Purpose: Among the materials for cranioplasty, autogenous bone is ideal because it is less susceptible to infection and has lower rates of subsequent exposure. However, the procedure is technically demanding to perform and requires a donor site. Disadvantages further exist when the defect is large and there are attendant limitations in donor site. The authors present their experience with reconstruction of large skull defect using right-angled zigzag osteotomized outer table of autogenous calvarial bone, overcoming the limitation in donor site. Methods: From 2000 to 2006, 9 patients were retrospectively reviewed, who had undergone reconstruction with right angled zigzag osteotomized outer table of autogenous calvarial bone. Results: Aesthetically satisfactory skull shape was achieved. Major complications of infection, hematoma, plate exposure, and donor site complications of dural tear with bleeding, cerebrospinal fluid leak, and meningitis were not seen. One patient had delayed wound healing and was successfully managed conservatively. Conclusion: Autogenous bone is the material of choice for cranioplasty, especially in complicated cases. Right angled zigzag osteotomy is a useful method in reconstruction of large skull defects with less donor site morbidity.
Introduction : The purpose of this study was to evaluate the possibility of the acellular dermal matrix (ADM) as a barrier membrane for bone regeneration, and to evaluate the osteogenic effect of ADM as a carrier system for rhBMP-2 in the rat calvarial defect model. Materials and Methods: An 8-mm, calvarial, critical-size osteotomy defect was created in each of 60 male Spraque-Dawley rats(weight $250{\sim}300g$). Three groups of 20 animals, each received either rhBMP-2(0.025mg/ml) in an ADM carrier, ADM only, or negative surgical control. And each group was divided into 2- and 8-weeks healing intervals. The groups were evaluated by histologic and histomorphometric parameters(10 animals/group/healing intervals). Data were expressed as $means{\pm}standard$ deviations($m{\pm}SD$). Comparisons between experimental and control groups were made using two-way ANOVA and post hoc t-test. Comparisons between 2 weeks and 8 weeks were made using paired t-test. The level of statistical difference was defined as P< 0.05. Results : The ADM group and rhBMP-2/ADM group results in enhanced local bone formation in the rat calvarial defect at both 2 and 8 weeks. The amount of defect closure and new bone formation were significantly greater in the rhBMP-2/ADM group relative to ADM group(P<0.05). At 8 weeks, the majority of ADM in the defect was contracted, and integrated with surrounding host tissues. In addition, host cell infiltration and neovascularization of the ADM in the absence of an inflammatory response were observed, and the newly formed bone around ADM showed a continuous remodeling and consolidation. Conclusion : The results of the present study indicated that ADM may be used as a barrier membrane for bone regeneration and that may be employed as a delivery system for BMPs.
Kim, Mi-Suk;Yeo, Hwan-Ho;Kim, Su-Gwan;Lim, Sung-Chul
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.28
no.4
/
pp.274-279
/
2002
The purpose of this study is to evaluate the critical maintenance period of absorbable membrane for guided bone regeneration. Fortynine Sprague-Dawley rats weighing about 300g were divided into seven groups. An 8 mm circular full-thickness defect in calvarial bone was made and then cellular acetate porous filter (Millipore $filter^{(R)}$.) was placed on the calvarial bone defect. The filter was removed at 2, 3, 4, 5, 6, 8 and 11 weeks after placement. Rats were sacrificed at 12 weeks the placement of cellular acetate porous filter. The specimens were stained with Hematoxylin-Eosin and observed under light microscope. The amount of regenerated bone was measured from both margin of calvarial bone defect (unit : mm). The results were as follows. Bone regeneration of each experimental group was increased gradually and the bond defect was almost completely filled with new bone in 5-, 6-, 8-, and 11-week experimental group. Histologic findings showed mild inflammatory response and granulation tissue formation without apparent adverse effects on the healing process. In 11-week experimental group, the bone defect was completely filled with new bone containing abundant osteoid which was oriented to the dural side and contribute to bony thickening. We suggest that non-absorbable membrane and bioabsorbable membrane presumably should remain intact for longer than 5 weeks to be effective.
Purpose: The aim of this study was to assess the possible paracrine effect of bone morphogeneticprotein-2 (BMP-2) at the experimental site on the adjacent control site for validating a rabbit calvarial defect model as a means of verifying the effect of BMP-2. Methods: Sixteen rabbits were divided into two groups (n=8 in each) according to whether or not BMP-2 would be used. Two circular defects (8 mm in diameter) were created side by side, 2 mm apart, in the calvarium of all of the rabbits. In each animal, one of the defects was grafted with either BMP-2-loaded carrier or carrier material alone. The control defects adjacent to these grafted defects, designated CB (the nongrafted defect adjacent BMP-2-loaded carrier-grafted defect) and CC (the nongrafted defect adjacent to carrier only-grafted defect), respectively, were the focus of this study, and were filled only with a blood clot in all of the animals. Histologic observation and histomorphometric analysis were performed at 2 and 8 weeks (n=4 animals per point in time) after surgery. Results: There was no noteworthy difference in the healing pattern, and no statistically significant differences in histomorphometric parameters such as the defect closure, new bone area, or total augmented area between the CC and CB groups. Conclusions: The results of this study suggest that rabbit calvarial defects separated by a distance of 2 mm are suitable for evaluating the effects of BMP-2 and the control defect can be regarded not to be affected by BMP-2 applied defect.
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