• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.44 no.2
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• pp.79-85
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• 2018

Objectives: The aim of this study was to evaluate the effects of herbal extracts on bone regeneration. Two known samples were screened. Materials and Methods: We previously established a rat calvaria defect model using a combination of collagen scaffold and herbal extracts. An 8 mm diameter trephine bur with a low-speed dental hand piece was used to create a circular calvaria defect. The experimental group was divided into 4 classifications: control, collagen matrix, Danshen with collagen, and Ge Gan with collagen. Animals in each group were sacrificed at 4, 6, 8, and 10 weeks after surgery, and bone regeneration ability was evaluated by histological examination. Results: Results revealed that both Danshen and Ge Gan extracts increased bone formation activity when used with collagen matrix. All groups showed almost the same histological findings until 6 weeks. However, after 6 weeks, bone formation activity proceeded differently in each group. In the experimental groups, new bone formation activity was found continuously up to 10 weeks. In the Danshen and Ge Gan groups, grafted materials were still present until 10 weeks after treatment, as evidenced by foreign body reactions showing multinucleated giant cells in chronic inflammatory vascular connective tissue. Conclusion: Histological analyses showed that Danshen and Ge Gan extractions increased bone formation activity when used in conjunction with collagen matrix.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.33 no.2
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• pp.112-119
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• 2011

Purpose: Collagen membranes are used extensively as bioabsorbable barriers in guided bone regeneration. However, collagen has different effects on tissue restoration depending on the type, structure, degree of cross-linking and chemical treatment. The purpose of this study was to evaluate the inflammatory reaction, bone formation, and degradation of dehydrothermal treated porcine type I atelocollagen (CollaGuide$^{(R)}$) compared to of the non-crosslinked porcine type I, III collagen (BioGide$^{(R)}$) and the glutaldehyde cross-linked bovine type I collagen (BioMend$^{(R)}$) in surgically created bone defects in rat mandible. Methods: Bone defect model was based upon 3 mm sized full-thickness transcortical bone defects in the mandibular ramus of Sprague-Dawley rats. The defects were covered bucolingually with CollaGuide$^{(R)}$, BioMend$^{(R)}$, or BioGide$^{(R)}$ (n=12). For control, the defects were not covered by any membrane. Lymphocyte, multinucleated giant cell infiltration, bone formation over the defect area and membrane absorption were evaluated at 4 weeks postimplantation. For comparison of the membrane effect over the bone augmentation, rats received a bone graft plus different covering of membrane. A $3{\times}4$ mm sized block graft was harvested from the mandibular angle and was laid and stabilized with a microscrew on the naturally existing curvature of mandibular inferior border. After 10 weeks postimplantation, same histologic analysis were done. Results: In the defect model at 4 weeks post-implantation, the amount of new bone formed in defects was similar for all types of membrane. Bio-Gide$^{(R)}$ membranes induced significantly greater inflammatory response and membrane resorption than other two membranes; characterized by lymphocytes and multinucleated giant cells. At 10 weeks postoperatively, all membranes were completely resorbed. Conclusion: Dehydrotheramal treated cross-linked collagen was safe and effective in guiding bone regeneration in alveolar ridge defects and bone augmentation in rats, similar to BioGide$^{(R)}$ and BioMend$^{(R)}$, thus, could be clinically useful.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.8
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• pp.1390-1393
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• 2003

Measuring in vivo rate of bone collagen synthesis has so far been technically difficult and often subject to quite large errors. In the present study, bone collagen synthesis rate was measured using a precursor-product method, based on the exchange of $^2$$H_2O$ into amino acids. Mass isotopomer abundance in hydroxyproline from bone collagen was analyzed by gas chromatography/mass spectrometry. The $^2$$H_2O$ labeling protocol consisted of an initial intraperitoneal injection of 99.9% $^2$$H_2O$, to achieve approximately 2.5% body water enrichment followed by administration of 4% $^2$$H_2O$ in drinking water for 9 weeks. Body $^2$$H_2O$ enrichments were stable at 2.7 ∼ 3.0% over labeling Period. In growing rats, the fractional synthesis rate ( $k_{s}$) of bone collagen was 0.066 $\pm$ 0.049 w $k^{-1}$ . The unique features of stable $^2$$H_2O$ pools and label incorporation allowed the precursor-product approach to be used for measuring bone collagen synthesis rate..

• Journal of Periodontal and Implant Science
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• v.34 no.1
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• pp.223-241
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• 2004

The purpose of this present study evaluated the osseous response around Ca-P coated xenogenic bone and compared osteogenic potential of Ca-P coated xenogenic bone to that of combination with type I collagen derived from bovine tendon as a biocompatible binder to prevent migration of bone particle on the repair of calvarial defects in rabbits. To study the effects of Ca-P coated xenogenic bone and collagen on bone healing, four 5-mm-diameter skull defect were made in calvaria with trephine filled with an autogenous bone chip or Ca-P coated xenogenic bone or Ca-P coated xenogenic bone and type I collagen (1:1 mixture by volume) or left empty. The defects were evaluated histologically at 1, 2, 4 and 8 weeks following implantation. Ca-P coated xenogenic bone at the calvarial defects of rabbits showed osteoconductivity at the margin of defect in the early stage of bony healing, but no direct contact with new bone was observed. With time passed by, it was resorbed slowly and showed consistent inflammatory reaction. An additional use of type I collagen derived from bovine tendon improved clinical handling, but no new bone formation was observed histologically. Above all, autogenous bone graft showed most prominent healing in quantity and density of new bone formation. According to this study, the use of Ca-P coated xenogenic bone alone and combination with type I collagen did not showed effective healing in quantity and density of new bone formation.

• Proceedings of the Materials Research Society of Korea Conference
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• 2011.05a
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• pp.54.2-54.2
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• 2011

DTBP (dimethyl 3,3`-dithiobispropionimidate) was applied to collagen and gelatin coating on BCP granules and a crosslinking agent. The DTBP crosslinking was done for decreasing the solubility of the coating and hence increasing the stability. The nanostructure of collagen and gelatin coating surfaces were observed by SEM technique. Based on the DSC thermograms and FT-IR spectrums, the crosslinkings were confirmed between collagen molecules and gelatin molecules. The compressive strength was measured before crosslinking and after that. In-vitro study was carried out by measuring cell viability and observing cell morphology after DTBP crosslinking. Moreover, the proliferation ability of MG-63 osteoblast-like cells on the crosslinked BCP granules was evaluated by Western blot assay. The BCP granules were implanted into rabbit femur for 4 weeks and 12 weeks. The bone tissue formation was analyzed with micro-computed tomography (micro-CT) and histological analysis was also carried out by hematoxylin and eosin (H&E) staining for visualization of cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.2
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• pp.324-330
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• 2004

The purpose of this study was to investigate the effects of Porphyra tenera (PT) extracts on formation of 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 PT at 50 mg/kg bw/day (OVX-PT5O), 200 mg/kgbw/day (OVX-PT200). The PT ethanol extracts were orally administrated 1 mL per day. Body weight gain, food intake and food efficiency ratio were significantly different among the groups. The change of collagen content was studied in lung, bone, cartilage and skin of ovariectomized rats. The effects of PT extracts on the amount of collagen were examined by measuring the hydroxyproline, which is a specific amino acid existing in collagen. Pyridinoline is pyridinium cross-link formed in the mature form of collagen from lysine and hydroxylysine residues. Pyridinoline content was analyzed by HPLC. Pyridinoline content in bone collagen was decreased by ovariectomy but supplementation with the PT extracts was similarly increased to Sham. These results suggest that the PT supplementation could decrease bone loss in postmenopausal women.

• Journal of Biomedical Engineering Research
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• v.9 no.2
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• pp.251-252
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• 1988

To develop an artificial bone substitute that is gradually degraded and replaced by the regenerated natural bone, the authors designed a composite that is consisted of calcium phosphate and collagen. To use as the structural matrix of the composite, collagen was purified from human umbilical cord. The obtained collagen was treated by pepsin to remove telopeptides, and finally, the immune-free atelocollagen was produced: The cross linked atelocollagen was highly resistant to the collagenase induced collagenolysis. The cross linked collagen demonstrated an improved tensile strength.

• Journal of Periodontal and Implant Science
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• v.49 no.4
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• pp.258-267
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• 2019

Purpose: Increased bone regeneration has been achieved through the use of stem cells in combination with graft material. However, the survival of transplanted stem cells remains a major concern. The purpose of this study was to evaluate the viability of transplanted mesenchymal stem cells (MSCs) at an early time point (24 hours) based on the type and form of the scaffold used, including type I collagen membrane and synthetic bone. Methods: The stem cells were obtained from the periosteum of the otherwise healthy dental patients. Four symmetrical circular defects measuring 6 mm in diameter were made in New Zealand white rabbits using a trephine drill. The defects were grafted with 1) synthetic bone (${\beta}$-tricalcium phosphate/hydroxyapatite [${\beta}-TCP/HA$]) and $1{\times}10^5MSCs$, 2) collagen membrane and $1{\times}10^5MSCs$, 3) ${\beta}-TCP/HA+collagen$ membrane and $1{\times}10^5MSCs$, or 4) ${\beta}-TCP/HA$, a chipped collagen membrane and $1{\times}10^5MSCs$. Cellular viability and the cell migration rate were analyzed. Results: Cells were easily separated from the collagen membrane, but not from synthetic bone. The number of stem cells attached to synthetic bone in groups 1, 3, and 4 seemed to be similar. Cellular viability in group 2 was significantly higher than in the other groups (P<0.05). The cell migration rate was highest in group 2, but this difference was not statistically significant (P>0.05). Conclusions: This study showed that stem cells can be applied when a membrane is used as a scaffold under no or minimal pressure. When space maintenance is needed, stem cells can be loaded onto synthetic bone with a chipped membrane to enhance the survival rate.

• Journal of Korean Dental Science
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• v.14 no.1
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• pp.12-25
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• 2021

Purpose: (i) To evaluate the biologic properties of a bi-layered 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride-cross-linked collagen membrane (CCM) in vitro. (ii) To assess the efficacy of CCM for localized bone regeneration in vivo. Materials and Methods: Biodegradation of CCM compared to a native collagen membrane (NCM) was assessed in vitro. In vivo, twelve male New Zealand White rabbits were used. Four calvarial, circular defects (diameter 8 mm) were created in each animal. The sites were randomly allocated to i) CCM+biphasic calcium phosphate (BCP) (CCM-BCP group), ii) CCM alone (CCM), iii) BCP alone (BCP) and, iv) negative control (control). Animals were sacrificed at 2 (n=6) and 8 weeks (n=6). Outcome measures included: micro-computed tomography (μCT) analysis (total augmented volume [TAV], new bone volume) and histomorphometry (total augmented area [TAA], newly formed bone, remaining membrane thickness [RMT]). Result: CCM was more resistant to degradation than NCM. μCT analysis showed CCM-BCP (196.43±25.30 mm³) and BCP (206.23±39.13 mm³) groups had significantly (P<0.01) larger TAV than the control (149.72±12.28 mm³) after 8 weeks. Histomorphometrically, CCM-BCP group (17.75±5.97 mm²) had significantly (P<0.01) greater TAA compared to the CCM group (7.74±2.25 mm²) and the control (8.13±1.81 mm²) after 8 weeks. After 8 weeks, RMT was reduced by 67%. Conclusion: CCM can be a favorable choice of barrier membrane when performing guided bone regeneration (GBR) in localized bone defects. CCM has better resistance to degradation than the natural collagen membrane, in vitro. In vivo, CCM provides an advantageous integration of prolonged barrier function and biocompatibility for GBR.

• Journal of Korean Dental Science
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• v.15 no.1
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• pp.51-60
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• 2022

Purpose: The aim of this study was to evaluate the bio-durability and bone regeneration capacity of the non-cross-linked collagen membrane in rabbit calvarial defect models. Materials and Methods: Four circular defects with 8 mm diameter were made in each of calvarium of 10 male rabbits. The following groups was randomly assigned to each defect - 1) Control, 2) membrane group containing non-cross-linked collagen membrane only (M), 3) bone graft group (B), 4) bone graft with membrane group (B+M). Animals were sacrificed and samples were harvested at 2 weeks (n=5) and 8 weeks (n=5). Histologic sections were prepared and histomorphometric analysis was performed. Result: Histologic results showed well adaptation of the non-cross-linked membrane on each defect and normal healing response at 2 weeks. At 8 weeks, the membranes were partially biodegraded. Histomorphometrically, B and B+M group showed the significantly greater total augmented area (B+M group, 10.44±1.49, P=0.016; B group, 9.13±0.53, P=0.032) and new bone formation (B+M group, 2.89±0.93, P=0.008; B group, 2.85±1.15, P=0.008) compared to control group. Collapsing of the central portion of the membrane, membrane group showed greater value in new bone formation at 8 weeks (1.78±0.68, P=0.032). Conclusion: Within the limitations of this study, the non-cross-linked collagen membrane fabricated using the improved decellularized method was shown to be effective for the regeneration of calvarial bone defects. In addition, prolonged barrier function might be provided using this collagen membrane.