• Journal of Periodontal and Implant Science
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• v.53 no.6
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• pp.467-477
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• 2023

Purpose: The purpose of this study was to evaluate the regenerative capacity of stem cells combined with bone graft material and a collagen matrix in rabbit calvarial defect models according to the type and form of the scaffolds, which included type I collagen matrix and synthetic bone. Methods: Mesenchymal stem cells (MSCs) were obtained from the periosteum of participants. Four symmetrical 6-mm-diameter circular defects were made in New Zealand white rabbits using a trephine drill. The defects were grafted with (1) group 1: synthetic bone (β-tricalcium phosphate/hydroxyapatite [β-TCP/HA]) and 1×10⁵ MSCs; (2) group 2: collagen matrix and 1×10⁵ MSCs; (3) group 3: β-TCP/HA, collagen matrix covering β-TCP/HA, and 1×10⁵ MSCs; or (4) group 4: β-TCP/HA, chipped collagen matrix mixed with β-TCP/HA, and 1×10⁵ MSCs. Cellular viability and cell migration rates were analyzed. Results: Uneventful healing was achieved in all areas where the defects were made at 4 weeks, and no signs of infection were identified during the healing period or at the time of retrieval. New bone formation was more evident in groups 3 and 4 than in the other groups. A densitometric analysis of the calvarium at 8 weeks post-surgery showed the highest values in group 3. Conclusions: This study showed that the highest regeneration was found when the stem cells were applied to synthetic bone along with a collagen matrix.

• Journal of Veterinary Clinics
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• v.39 no.2
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• pp.59-64
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• 2022

A dog aged two years and seven months and a cat aged seven years were referred owing to fractures of long bones. Preoperative radiographs revealed comminuted bone fractures close to joints. Conventionally, long-bone fractures are treated using intramedullary pins, plate and screw systems, or an external fixator system. In cases of non-reducible fractures, various graft materials have been used in fracture treatments to stimulate bone repair. Here, recombinant human bone morphogenetic protein-2 (rhBMP-2) and a collagen membrane were applied. Four weeks after surgery, fractured bone fragments began to unite and the bone union was observed using radiography four months after surgery. No complications occurred related to grafted materials. We successfully applied rhBMP-2 and collagen membranes in two different species to support the healing process of comminuted fractures, according to the concept of guided bone regeneration.

• Preventive Nutrition and Food Science
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• v.16 no.4
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• pp.291-298
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• 2011

Yam extracts (Dioscorea batatas) have been reported to possess a variety of functions. However, studies on its osteogenic properties are limited. In this study, we investigated the effect of ethanol and water extracts on osteoblast proliferation and bone matrix protein synthesis, type I collagen and alkaline phosphatase (ALP), using osteoblastic MC3T3-E1 cell model. MC3T3-E1 cells were cultured with yam ethanol and water extracts (0~30 mg/L) within 39 days of osteoblast differentiation period. Cell proliferation was measured by MTT assay. Bone matrix proteins were assessed by the accumulation of type I collagen and ALP activity by staining the cell layers for matrix staining. Also, the secreted (media) matrix protein concentration (type I collagen) and enzyme activity (ALP) were measured colorimetrically. Yam ethanol and water extracts stimulated cell proliferation within the range of 15~30 mg/L at 15 day treatment. The accumulation of type I collagen in the extracellular matrix, as well as secreted collagen in the media, increased with increasing doses of yam ethanol (3~15 mg/L) and water (3~30 mg/L) extracts. ALP activity was not affected by yam ethanol extracts. Our results demonstrated that yam extracts stimulated osteoblast proliferation and enhanced the accumulation of the collagenous bone matrix protein type I collagen in the extracellular matrix. These results suggest that yam extracts may be a potential activator for bone formation by increasing osteoblast proliferation and increasing bone matrix protein type I collagen. Before confirming the osteogenic action of yam, further studies for clarifying how and whereby yam extracts can stimulate this ostegenesis action are required.

• Journal of Periodontal and Implant Science
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• v.49 no.2
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• pp.90-104
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• 2019

Purpose: The aim of this study was to evaluate clinical and radiographic changes and the survival rate after periodontal surgery using deproteinized bovine bone mineral (DBBM) with 10% collagen or DBBM with a collagen membrane in endo-periodontal lesions. Methods: A total of 52 cases (41 patients) with at least 5 years of follow-up were included in this study. After scaling and root planing with or without endodontic treatment, periodontal regenerative procedures with DBBM with 10% collagen alone or DBBM with a collagen membrane were performed, yielding the DBBM + 10% collagen and DBBM + collagen membrane groups, respectively. Changes in clinical parameters including the plaque index, bleeding on probing, probing pocket depth, gingival recession, relative clinical attachment level, mobility, and radiographic bone gains were evaluated immediately before periodontal surgical procedures and at a 12-month follow-up. Results: At the 12-month follow-up after regenerative procedures, improvements in clinical parameters and radiographic bone gains were observed in both treatment groups. The DBBM + 10% collagen group showed greater probing pocket depth reduction ($4.52{\pm}1.06mm$) than the DBBM + collagen membrane group ($4.04{\pm}0.82mm$). However, there were no significant differences between the groups. Additionally, the radiographic bone gain in the DBBM + 10% collagen group ($5.15{\pm}1.54mm$) was comparable to that of the DBBM + collagen membrane group ($5.35{\pm}1.84mm$). The 5-year survival rate of the teeth with endo-periodontal lesions after periodontal regenerative procedures was 92.31%. Conclusions: This study showed that regenerative procedures using DBBM with 10% collagen alone improved the clinical attachment level and radiographic bone level in endo-periodontal lesions. Successful maintenance of the results after regenerative procedures in endo-periodontal lesions can be obtained by repeated oral hygiene education within strict supportive periodontal treatment.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.34 no.1
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• pp.36-48
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• 2008

To repair bone defects in the oral and maxillofacial field, bone grafts including autografts, allografts, and artificial bone are used in clinical dentistry despite several disadvantages. The purpose of this study was to evaluate new bone formation and healing in rat calvarial bone defects using hydroxyapatite (HA, $Ca_{10}[PO_4]_6[OH]_2,\;Bongros^{(R)}$, Bio@ Co., KOREA) and tricalcium phosphate (${\beta}-TCP,\;Ca_3[PO_4]_2$, Sigma-Aldrich Co., USA) mixed at various ratios. Additionally, this study evaluated the effects of type I collagen (Rat tail, BD Biosciences Co., Sweden) as a basement membrane organic matrix. A total of twenty, 8-week-old, male Sprague-Dawley rats, weighing 250-300g, were divided equally into a control group (n=2) and nine experimental groups (n=2, each). Bilateral, standardized transosseous circular calvarial defects, 5.0 mm in diameter, were created. In each experimental group, the defect was filled with HA and TCP at a ratio of 100:0, 80:20, 70:30, 60:40, 50:50, 40:60, 30:70, 20:80, and 0:100 with or without type I collagen. Rats were sacrificed 4 and 8 weeks post-operation for radiographic (standardized plain film, Kodak Co., USA), histomorphologic (H&E [Hematoxylin and Eosin], MT [Masson Trichrome]), immunohistochemical staining (for BMP-2, -4, VEGF, and vWF), and elementary analysis (Atomic absorption spectrophotometer, Perkin Elmer AAnalyst $100^{(R)}$). As the HA proportion increased, denser radiopacity was seen in most groups at 4 and 8 weeks. In general radiopacity in type I collagen groups was greater than the non-collagen groups, especially in the 100% HA group at 8 weeks. No new bone formation was seen in calvarial defects in any group at 4 weeks. Bridging bone formation from the defect margin was marked at 8 weeks in most type I collagen groups. Although immunohistochemical findings with BMP-2, -4, and VEGF were not significantly different, marked vWF immunoreactivity was present. vWF staining was especially strong in endothelial cells in newly formed bone margins in the 100:0, 80:20, and 70:30 ratio type I collagen groups at 8 weeks. The calcium compositions from the elementary analysis were not statistically significant. Many types of artificial bone have been used as bone graft materials, but most of them can only be applied as an inorganic material. This study confirmed improved bony regeneration by adding organic type I collagen to inorganic HA and TCP mixtures. Therefore, these new artificial bone graft materials, which are under strict storage and distribution systems, will be suggested to be available to clinical dentistry demands.

• Journal of Periodontal and Implant Science
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• v.51 no.1
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• pp.40-52
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• 2021

Purpose: Various crosslinking methods have been introduced to increase the longevity of collagen membranes. The aim of this study was to compare and evaluate the degradation and bone regeneration patterns of 3 collagen membranes. Methods: Four 8-mm-diameter circular bone defects were created in the calvaria of 10 rabbits. In each rabbit, each defect was randomly allocated to 1) the sham control group, 2) the non-crosslinked collagen sponge (NS) group, 3) the chemically crosslinked collagen membrane (CCM) group, or 4) the biphasic calcium phosphate (BCP)-supplemented ultraviolet (UV)-crosslinked collagen membrane (UVM) group. Each defect was covered with the allocated membrane without any graft material. Rabbits were sacrificed at either 2 or 8 weeks post-surgery, and radiographic and histologic analyses were done. Results: New bone formed underneath the membrane in defects in the CCM and UVM groups, with a distinctive new bone formation pattern, while new bone formed from the base of the defect in the NS and control groups. The CCM maintained its shape until 8 weeks, while the UVM and NS were fully degraded at 8 weeks; simultaneously, sustained inflammatory infiltration was found in the margin of the CCM, while it was absent in the UVM. In conclusion, the CCM showed longer longevity than the UVM, but was accompanied by higher levels of inflammation. Conclusions: Both the CCM and UVM showed distinctive patterns of enhancement in new bone formation in the early phase. UV crosslinking can be a biocompatible alternative to chemical crosslinking.

• Journal of Periodontal and Implant Science
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• v.48 no.4
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• pp.236-250
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• 2018

Purpose: Resorption of the alveolar bone is an unavoidable consequence of tooth extraction when appropriate alveolar ridge preservation (ARP) measures are not taken. The objective of this trial was to test the hypothesis that dimensional changes in the alveolar bone after tooth extraction would be reduced by inserting an equine collagen membrane and a collagen cone to fill and seal the alveolus (as ARP), in comparison to extraction with untreated alveoli. Methods: In this randomized clinical trial, 31 patients were directly treated with the collagen material after extraction of a tooth from the maxilla (the ARP group). Twenty-nine patients served as the control group. After extraction, no further treatment (i.e., no socket preservation measures) was performed in the control group. Changes in the alveolar process immediately after extraction and after an 8 (${\pm}1$)-week healing period were evaluated 3-dimensionally. Blinded analyses were performed after superimposing the data from the digitalized impressions and surfaces generated by cone-beam computed tomography. Results: Both the ARP and control groups showed a reduction of bone in the alveolar area after tooth extraction. However, significantly less bone resorption was detected in the clinically relevant buccal region in the ARP group. The median bone reduction was 1.18 mm in the ARP group and 5.06 mm in the control group (P=0.03). Conclusions: The proposed hypothesis that inserting a combination material comprising a collagen cone and membrane would lead to a difference in alveolar bone preservation can be accepted for the clinically relevant buccal distance. In this area, implantation of the collagen material led to significantly less alveolar bone resorption. German Clinical Trials Register at www.drks.de, DRKS00004769.

• Journal of Periodontal and Implant Science
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• v.50 no.1
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• pp.14-27
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• 2020

Purpose: To overcome several drawbacks of chemically-crosslinked collagen membranes, modification processes such as ultraviolet (UV) crosslinking and the addition of biphasic calcium phosphate (BCP) to collagen membranes have been introduced. This study evaluated the efficacy and biocompatibility of BCP-supplemented UV-crosslinked collagen membrane for guided bone regeneration (GBR) in a rabbit calvarial model. Methods: Four circular bone defects (diameter, 8 mm) were created in the calvarium of 10 rabbits. Each defect was randomly allocated to one of the following groups: 1) the sham control group (spontaneous healing); 2) the M group (defect coverage with a BCP-supplemented UV-crosslinked collagen membrane and no graft material); 3) the BG (defects filled with BCP particles without membrane coverage); and 4) the BG+M group (defects filled with BCP particles and covered with a BCP-supplemented UV-crosslinked collagen membrane in a conventional GBR procedure). At 2 and 8 weeks, rabbits were sacrificed, and experimental defects were investigated histologically and by micro-computed tomography (micro-CT). Results: In both micro-CT and histometric analyses, the BG and BG+M groups at both 2 and 8 weeks showed significantly higher new bone formation than the control group. On micro-CT, the new bone volume of the BG+M group (48.39±5.47 ㎣) was larger than that of the BG group (38.71±2.24 ㎣, P=0.032) at 8 weeks. Histologically, greater new bone area was observed in the BG+M group than in the BG or M groups. BCP-supplemented UV-crosslinked collagen membrane did not cause an abnormal cellular reaction and was stable until 8 weeks. Conclusions: Enhanced new bone formation in GBR can be achieved by simultaneously using bone graft material and a BCP-supplemented UV-crosslinked collagen membrane, which showed high biocompatibility and resistance to degradation, making it a biocompatible alternative to chemically-crosslinked collagen membranes.

• Journal of Life Science
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• v.18 no.11
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• pp.1578-1583
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• 2008

The purpose of this study was to investigate the effects of sea tangle (ST) extracts on formation of collagen and collagen cross-link in ovariectomized rats. From day 3 until 42 after the ovariectomy, Sprague-Dawley female rats were randomly assigned to the following groups: sham-operated rats(Sham), ovariectomized control rats (OVX-control), ovariectomized rats supplemented with ST at 50 mg/kg bw/day (OVX-ST50), 200 mg/kg bw/day (OVX-ST200). The ethanol extraxcr of ST was orally administrated at 1 ml/day. The change of collagen content was investigated in bone, cartilage and skin of ovariectomized rats. Effects of ST extract on the amount of collagen was examined by measuring the hydroxyproline, which is a specific amino acid existing in collagen. The ovariectomy resulted in a decrease in the levels of collagen content in bone and cartilage tissues. However, the supplementation with the sea tangle extract prevented the decrease in the collagen level in bone and cartilage tissues. Pyridinoline is pyridinium cross-link formed in the mature form of collagen from lysine and hydroxylysine residues. Although the pyridinoline content in bone collagen declined after ovariectomy, it was recovered to a normal level of Sham group by the supplementation with the ST extracts. In addition, the deoxypyridinoline content in bone collagen, which was reduced after ovariectomy was enhanced to normal level by the supplementation with the ST extract. These results was consistent with the conclusions based on estrogenic activities of ST.

• BMB Reports
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• v.47 no.2
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• pp.110-114
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• 2014

Fibrin is a natural provisional matrix found in wound healing, while type I collagen is a major organic component of bone matrix. Despite the frequent use of fibrin and type I collagen in bone regenerative approaches, their comparative efficacies have not yet been evaluated. In the present study, we compared the effects of fibrin and collagen on the proliferation and differentiation of osteoblasts and protein adsorption. Compared to collagen, fibrin adsorbed approximately 6.7 times more serum fibronectin. Moreover, fibrin allowed the proliferation of larger MC3T3-E1 pre-osteoblasts, especially at a low cell density. Fibrin promoted osteoblast differentiation at higher levels than collagen, as confirmed by Runx2 expression and transcriptional activity, alkaline phosphatase activity, and calcium deposition. The results of the present study suggest that fibrin is superior to collagen in the support of bone regeneration.