Decellularized Non-cross-linked Collagen Membranes for Guided Bone Regeneration in Rabbit Calvarial Defects

Purpose@#The aim of this study was to evaluate the bio-durability and bone regeneration capacity of the non-crosslinked 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 noncross-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.

Distinctive bone regeneration of calvarial defects using biphasic calcium phosphate supplemented ultraviolet-crosslinked collagen membrane

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 mm3) was larger than that of the BG group (38.71±2.24 mm3, 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.

Bioactive characteristics of an implant surface coated with a pH buffering agent: an in vitro study

PURPOSE: The purpose of this study was to evaluate the effectiveness of conventional sandblasted, large-grit, acid-etched (SLA) surface coated with a pH buffering solution based on surface wettability, blood protein adhesion, osteoblast affinity, and platelet adhesion and activation.METHODS: Titanium discs and implants with conventional SLA surface (SA), SLA surface in an aqueous calcium chloride solution (CA), and SLA surface with a pH buffering agent (SOI) were prepared. The wetting velocity was measured by the number of threads wetted by blood over an interval of time. Serum albumin adsorption was tested using the bicinchoninic acid assay and by measuring fluorescence intensity. Osteoblast activity assays (osteoblast adhesion, proliferation, differentiation, mineralization, and migration) were also performed, and platelet adhesion and activation assays were conducted.RESULTS: In both the wetting velocity test and the serum albumin adsorption assay, the SOI surface displayed a significantly higher wetting velocity than the SA surface (P=0.000 and P=0.000, respectively). In the osteoblast adhesion, proliferation, differentiation, and mineralization tests, the mean values for SOI were all higher than those for SA and CA. On the osteoblast migration, platelet adhesion, and activation tests, SOI also showed significantly higher values than SA (P=0.040, P=0.000, and P=0.000, respectively).CONCLUSIONS: SOI exhibited higher hydrophilicity and affinity for proteins, cells, and platelets than SA. Within the limits of this study, it may be concluded that coating an implant with a pH buffering agent can induce the attachment of platelets, proteins, and cells to the implant surface. Further studies should be conducted to directly compare SOI with other conventional surfaces with regard to its safety and effectiveness in clinical settings.

Biomimetic characteristics of mussel adhesive protein-loaded collagen membrane in guided bone regeneration of rabbit calvarial defects

PURPOSE: The aim of the present study was to evaluate the biocompatibility and barrier function of mussel adhesive protein (MAP)-loaded collagen membranes in guided bone regeneration (GBR). METHODS: Eight male New Zealand white rabbits were used. Four circular defects (diameter: 8 mm) were created in the calvarium of each animal. The defects were randomly assigned to 1) a negative control group, 2) a cyanoacrylate (CA)-loaded collagen membrane group (the CA group), 3) a MAP-loaded collagen membrane group (the MAP group), and 4) a group that received a polycaprolactone block with MAP-loaded collagen membrane (the MAP-PCL group). Specimens were harvested at 2 weeks (n=4) and 8 weeks (n=4) postoperatively for observational histology and histometric analysis. RESULTS: In the histologic analysis, MAP was completely absorbed without any byproducts. In contrast, some of the CA adhesive remained, showing an inflammatory reaction, at 8 weeks. In the MAP-PCL group, the MAP-loaded collagen membranes served as a barrier membrane despite their fast degradation in GBR. No significant difference was found in the amount of new bone between the MAP-PCL and MAP groups (1.82±0.86 mm2 and 2.60±0.65 mm2, respectively). CONCLUSIONS: The MAP-loaded collagen membrane functioned efficiently in this rabbit calvarial GBR model, with excellent biocompatibility. Further research is needed to assess clinical applications in defect types that are more challenging for GBR than those used in the current model.

Bone Regeneration Using Block-type Deproteinized Porcine Bone Mineral with Collagen Membrane Using 3,4-Dihydroxyphenylalanine as Bone Adhesive

PURPOSE: The purpose of this study was to assess the adhesiveness and cytotoxicity of 3, 4-dihydroxyphenylalanine (DOPA), and to evaluate the role of collagen membrane with DOPA in the guided bone regeneration. MATERIALS AND METHODS: Peel resistance and cell cytotoxicity test were performed. Four defect types in nine rabbit calvaria were randomly allocated: i) control, ii) membrane, iii) deproteinized porcine bone mineral (DPBM) covered by membrane with DOPA, and iv) DPBM covered by membrane with cyanoacrylate. Animals were sacrificed at 2 (n=4) and 8 weeks (n=5) for microcomputed tomography and histomorphometric analysis. DOPA showed low peel resistance but high cell viability. RESULT: Cyanoacrylate and DOPA groups showed significantly higher mineralized tissue volume (MTV) compared to control and membrane groups at 2 weeks (P < 0.05). At 8 weeks, DOPA group showed the highest MTV. Significantly higher new bone area was found in DOPA group at 8 weeks (P < 0.05). Bone formation increased from 2 to 8 weeks in DOPA group (P < 0.05). CONCLUSION: DOPA showed high cell viability and in vivo study revealed predictable performance in bone regeneration.

Impact of immediate and non-immediate provisionalization on the soft tissue esthetics of final restorations on immediately placed implants / 대한치과보철학회지

STATEMENT OF PROBLEM: Interpoximal papilla could be re-established without immeidate support with a provisional resotration following an immdiate implant placement. PURPOSE: Successful esthetic outcomes were reported utilizing immediate provisionalization following immediate implant placements. The aim of this study was to evaluate the soft tissue esthetics around immediately placed single tooth implant restorations with or without immediate provisional restorations. METHODS: A total of ten patients, who had a hopeless maxillary anterior tooth, were enrolled in this study. Screw retained provisional restorations were delivered to the randomly chosen 5 patients (immediate provisionalization group) on the day of immediate implant placement and maintained for about 5 months. For the remaining five patients (non-immediate provisionalization group), healing abutments were delivered on the day of surgery, replaced with screw retained provisional restorations approximately 3 months afterwards, and the provisional restorations were maintained for about 3 months. Digital photographs were taken at the delivery of final restorations in order to assess following variables; mesial papilla, distal papilla, soft tissue level, soft tissue contour and facial soft tissue prominence. The variables were compared to those of the contralateral natural tooth and scored by prosthodontists, periodontists, orthodontists and dental students. RESULTS: The immediate provisionalization group marked significantly higher scores on the following variables; soft tissue level and facial soft tissue prominence. In evaluating each variable, there were no notable differences in opinion between four dentist groups. CONCLUSION: Immediate provisionalization can be a treatment option to achieve superior soft tissue esthetics around immediately placed single implant restorations rather than non-immediate provisionalization approaches.

Clinical Effect of the Subantimicrobial Dose of Doxycycline(SDD) on the Chronic Periodontitis / 대한치주과학회지

Periodontal disease is a complex infectious disease caused by bacteria in the oral mucosa, which results in gingival inflammation, breakdown of periodontal tissues, bone resorption, and finally tooth loss. Mechanical plaque control methods-scaling and root planing are effective methods to stop the progression of such periodontal disease. It was reported that subantimicrobial dose of doxycycline(SDD) regimen could improve clinical conditions of periodontal tissues without causing the overgrowth of opportunistic organisms that was a typical antibiotic side effect. Therefore pharmacological therapy, used in conjunction with mechanical therapy could be considered a useful treatment modality in the treatment of chronic periodontal disease. In this study, 30 patients diagnosed as moderate to advanced chronic periodontitis were divided into 2 groups. In this double-blind, placebo-controlled study, the patients were administered 20mg doxycycline capsule or placebo capsule b.i.d. for 4months, after scaling and root planing. Clinical parameters-bleeding on probing, pocket depth and clinical attachment level were compared and evaluated between these groups at periods of first visit, 1 month, 2 months, 3 months, 4 months. The results were as follows ; 1. In case of moderate periodontitis, pocket depth showed significant reduction after treatment in both the control & experiment groups, when compared with the baseline values(p<0.01), but in case of advanced periodontitis, only the experiment group showed significant reduction after treatment when compared with the baseline values(p<0.05). Statistically significant reduction in pocket depth was observed in the experiment group compared to the control group(p<0.05). 2. In case of moderate periodontitis, clinical attachment level showed significant reduction after treatment in both the control & experiment groups, when compared with the baseline values(p<0.01), but in case of advanced periodontitis, only the experiment group showed significant reduction after treatment when compared with the baseline values(p<0.05). Statistically significant reduction in clinical attachment level was observed in the experiment group compared to the control group(p<0.05). 3. Bleeding on probing improved after treatment in both the groups. In case of moderate periodontitis, the experiment group showed statistically significant reduction of bleeding on probing when compared with the control group at 1 and 4 months after treatment(p<0.05). In case of advanced periodontitis, treatment resulted in statistically significant reduction of bleeding on probing in both the groups(p<0.05). These results indicate that the use of subantimicrobial dose of doxycycline is a useful supplement to mechanical treatment for periodontal patients in ameliorating the clinical parameters such as periodontal pocket, attachment level, and bleeding on probing.

The effect of LiF-maleic acid added calcium aluminate bone cement & CA-PMMA composite bone cement on the healing of calvarial defect6) / 대한치주과학회지

The purpose of this study was to evaluate histologically the effect of LiF-maleic acid added calcium aluminate(LM-CA) bone cement & CA-PMMA composite bone cement on the healing of calvarial defect in Sprague-Dawley rats. The critical size defects were surgically produced in the calvarial bone using the 8mm trephine bur. The rats were divided in three groups : In the control group, nothing was applied into the defect of each rat. LM-CA bone cement was implanted in the experimental group 1 and CA-PMMA composite bone cement was implanted in the experimental group 2. Rats were sacrificed at 2, 8 weeks after surgical procedure. The specimens were examined by histologic analysis, especially about the bone-cement interface and the response of surrounding tissue. The results are as follows; 1. In the control group, inflammatory infiltration was observed at 2 weeks. At 8 weeks, periosteum and dura mater were continuously joined together in the defect area. But the center of defect area was filled up with the loose connective tissue. 2. In the experimental group 1, the bonding between implanted bone cement and the existing bone was seen, which more increased in 8 weeks than 2 weeks. Inflammatory infiltration and the dispersion of implanted bone cement particles were seen in both 2 weeks and 8 weeks. 3. In the experimental group 2, implanted bone itself had a dimensional stability and no bonding between implanted bone cement and the existing bone was seen in both 2 weeks and 8 weeks. Implanted bone cement was encapsulated by fibrous connective tissue. In addition, inflammatory infiltration was seen around implanted bone cement. On the basis of these results, when LM-CA bone cement or CA-PMMA composite bone cement was implanted in rat calvarial defect, LM-CA bone cement can be used as a bioactive bone graft material due to ability of bonding to the existing bone and CA-PMMA can be used as a graft material for augmentation of bone-volume due to dimensional stability.

The effects of calcium aluminate cement according to particle sizes on calvarial bone defects in rats / 대한치주과학회지

This present study was carried out to find the effects of calcium aluminate cement(CaO, Al2O3, CAC), which has been developed with bio-compatibility and mechanical properties, in biological environments. Two different particle sizes of CAC - 3.5micrometer vs. 212~250micrometer which is recommended in periodontal bone grafting procedures- were filled in 8mm calvarial defect in Sprague-Dawley rat. The specimens were examined histologically, especially the bone-cement interface and the response of surrounding tissues. The results are as follows; 1. In the control group, inflammatory cells were observed at 2 weeks. At 8 weeks, periosteum and dura mater were continuously joined together in the defect areas. But in the center of defect area were filled up with the loose connective tissues. 2. In the experimental group 1(212micrometer~250micrometer particle), immature bone was formed and outermost layer was surrounded by osteoid layer at 2 weeks. Osteoblasts were arranged between immature bone and osteoid layer. And, osteoid layer was remained until 8 weeks after surgery. 3. In the experimental group 2, periosteum and dura mater lost its continuity at 2 weeks. Scattering of CAC particles and infiltration of inflammatory cells were observed, which this findings deepened at 8 weeks. The result of this study shows that when calvarial defects in white rats are filled with calcium aluminate cement of 212~250micrometer, the materials are to be bio-compatible in growth and healing on surrounding tissues. When further researches are fulfilled, such as direct bone adhesion and bone regeneration ability, it's possible that CAC could be applied to various periodontology fields in the future.

The Study of Implant Patient's Type and Implant Distribution / 대한치주과학회지

It has been approximately 40 years since Branemark first introduced osseo-integration for implants in the early 1960s. Unlike crown and bridge or denture treatment, implant treatment helps preserve existing bone and improve masticatory functions. Thus, the awareness of implant treatment has grown rapidly among dentists and patients alike in Korea, as it becomes a widely accepted treatment. The following results on patient type and implant distribution were compiled from 1814 implant cases of 640 patients treated at the periodontal dept. of Y University Hospital during 1992 to 2001. 1. There are no dissimilarities between men and women, with patients in their 40, 50s accounting for 49% of patients and 56% of implant treatments; the largest share of patients and implant treatments. 2. Mn. posterior area accounted for 59% of implant treatments followed by Mx. posterior area(21%), Mx anterior area(14%) and Mn anterior area(6%). 3. Partial edentulous patients treated by single crown and bridge-type prosthesis accounted for 98% and fully edentulous patient accounted for the remaining 2%. 4. The major cause of tooth loss is periodontal disease, followed by dental caries, trauma and congenital missing. Compared to women, men are more likely to suffer from tooth loss due to periodontal disease. Also, older people are more likely to suffer from tooth loss due to periodontal disease rather than dental caries. 5. The distribution of bone quality for maxillae was 52% for type III, followed by 28% for type II, 20% for type IV and 0% for type I. As for mandible, the distribution was 52% for type II, followed by 37% for type III, 7% for type IV and 4% for type I. 6. The distribution of bone quantity for maxillae was 49% for type C, followed by 34% for type B, 14% for type D, 3% for type A, and 0% for type E. As for mandible, the distribution was 52% for type B, followed by 39% for type C, 6% for type D, 3% for type A and 0% for type E. 7. The majority of implants were those of 10-14mm in length (80%) and regular diameter in width (79%). The results provided us with basic data on patient type, implant distribution, bone condition, etc. We wish that our results coupled with other research data helps assist in the further study for better implant success/survival rates, etc.

A Clinical Results of Subepithelial Connective Tissue Graft for Root Coverage / 대한치주과학회지

Exposed root surfaces can cause esthetic problems, hypersensitivity, and root caries. Numerous efforts have been tried to cover the recessed root surfaces, and various techniques have been developed and introduced. Among these, subepithelial connective tissue graft which shows high coverage rate in various researches, has the advantage of good color match, less discomfort to the donor site, rich vascularity, and high predictability. Following results were obtained after investigating 6 and 18 months post operatively, 98 cases of subepithelial connective tissue graft from 48 patients who underwent subepithelial connective tissue graft procedure in the department of periodontology, college of dentistry, Yonsei university. 1. The total average root coverage of Miller class I, II & III were 76.2?24% at 6 months follow-up and 75?25.2% at 18 months follow-up with no statistically significant difference between the follow-up periods.(p<0.05) 2. The percentage of teeth showing complete coverage were 41.9% at 6 months follow-up and 39.2% at 18 months follow-up. 3. At 6 months follow-up, Miller classification I showed 84.9?20.7%, class II showed 82.5?17.7%, and class III showed 62.3?24.5% of coverage. In class III recession, statistically significantly less root coverage was observed compared to class I & II.(p<0.05) 4. At 18 months follow-up, Miller classification I showed 92.2?13.5%, class II showed 84.3?17.4%, and class III showed 59.5?24.5% of coverage. In class III recession, statistically significantly less root coverage was observed compared to class I & II.(p<0.05) In conclusion, subepithelial connective tissue graft for class I and II recession can be used as a clinically predictable treatment modality for root coverage.

Clinical effects of combination anorganic bovine-derived hydroxyapatite matrix(ABM)/cell binding peptide (P-15) in periodontal intrabony defects / 대한치주과학회지

The ultimate goal of periodontal theraphy is the regeneration of periodontal tissue which has been lost due to destructive periodontal disease. To achieve periodontal regeneration, various kinds of methods have been investigated and developed, including guided tissue regeneration and bone graft. Bone graft can be catagorized into autografts, allografts, xenografts, bone substitutes. And materials of all types have different biological activity and the capacity for periodontal regeneration, but ideal graft material has not been developed that fits all the requirement of ideal bone graft material. Intensive research is underway to identity, purify, synthesize a variety biologic modulators that may enhance wound healing and regeneration of lost tissues in periodontal therapy. The present study evaluates the effects of ABM/P-15 on the periodontal regeneration in intrabony defects of human. We used thirty four 2-wall or 3-wall osseous defects in premolars and molars of chronic periodontitis patient that have more than 5mm pockets and more than 3mm in intrabony defect. 12 negative control group underwent flap procedure only, 11 positive control group received DFDBA graft with flap procedure, and 11 experimental group received ABM/P-15 graft with flap procedure. The changes of probing pocket depth, loss of attachment and bone probing depth following 6months after treatment revealed the following results: 1. The changes of probing pocket depth showed a statistically significant decrease between after scaling and 6months after treatment in negative control(2.0+/-0.9mm), positive control(3.0+/-0.9mm), and experimental group (3.4+/-1.5mm) (P<0.01). Significantly more reduction was seen in experimental group compared to negative control group (P<0.05). 2. The changes of loss of attachment showed a statistically significant decrease between after scaling and 6months after treatment in positive control(2.0+/-0.6mm), and experimental group (2.2+/-1.0mm) except negative control group(0.1+/-0.7mm) (P<0.01). Significantly more reduction was seen in both experimental and positive control group compared to negative control group (P<0.05). 3. The changes of bone probing depth showed a statistically significant decrease between after scaling and 6months after treatment in positive control(2.7+/-1.0mm), and experimental group (3.4+/-1.3mm) except negative control(0.1+/-0.9mm) (P<0.01). Significantly more reduction was seen in both experimental and positive control group compared to negative control group (P<0.05). The results suggest that the use of ABM/P-15 in the treatment of periodontal intrabony defects can reduce loss of attachment and bone probing depth more than flap operation only. It suggests that ABM/P-15 may be an effective bone graft material for the regeneration of periodontal tissue in intrabony defects.

A Retrospective Anaylsis of Root-Resected Teeth / 대한치주과학회지

The purpose of this study was to evaluate the clinical validity of multi-rooted teeth subjected to root-resection treatment. Over a period of 1-7 years, 60 root-resected molars in 59 patients were examined clinically and radiographically. All patients were periodically recalled once or twice a year. Root-resections were due to periodontal, more specifically furcal bone loss or marginal bone loss, in 34 cases and extensive dental carious destruction in 10 cases. In other cases, root-resections resulted from 6 cases of root fracture, 6 cases of periodontal-endodontic combined lesion, and 1 case of endodontic problem. Root-resection was carried out on 26 maxillary molars and 34 mandibular molars. The results are as follows; 1. 14 cases(23.3%) were considered failures. 8 cases(13.3%) of them occurred within the first year(8 cases, 13.3%), 4 cases(6.7%) between 1-3 years, and 2 cases(3.3%) between 4-7 years. 2. 8 cases(13.3%) were considered failures due to periodontal reasons, 3 cases(5%) due to root fracture, 2 cases(3.3%) due to endodontic problem, and 1 case(1.7%) due to prosthetic problem. 3. 37 cases(61.7%) showed up for the recall appointments, and the percentage of failures(13.5%) was lower compared with that of all patients(23.3%). The results of the present study indicate that the prognosis of root-resected teeth is favourable if attention is paid to the selection of proper case and to achieving optimal oral hygiene and periodic check up.

The effects of chitosan on the human periodontal ligament fibroblasts in vitro / 대한치주과학회지

Periodontal therapy has dealt primarily with attempts at arresting progression of disease, however, more recent techniques have focused on regenerating the periodontal ligament having the capacity to regenerate the periodontium. The effect of chitosan(poly-N-acetyl glucosaminoglycan), a carbohydrate biopolymer extracted from chitin, on periodontal ligament regeneration is of particular interest. The purpose of this study was to evaluate the effect of chitosan on the human periodontal ligament fibroblasts(hPDLFs) in vitro, with special focus on their proliferative properties by MTT assay, the synthesis of type I collagen by reverse transcription-polymerase chain reaction(RT-PCR) and the activity of alkaline phosphatase(ALP). Fibroblast populations were obtained from individuals with a healthy periodontium and cultured with alpha-MEM as the control group. The experimental groups were cultured with chitosan in concentration of 0.01, 0.1, 1, 2mg/ml. The results are as follows; 1. Chitosan-induced proliferative responses of hPDLFs reached a plateau at the concentration of 0.1mg/ml (p<0.05). 2. When hPDLFs were stimulated with 0.1mg/ml chitosan, mRNA expression of type I collagen was up-regulated. 3. When hPDLFs were stimulated with 0.1mg/ml chitosan, ALP activity was significantly upregulated(p<0.05). In summary, chitosan(0.1mg/ml) enhanced the type I collagen synthesis in the early stage, and afterwards, facilitated differentiation into osteogenic cells. The results of this in vitro experiment suggest that chitosan potentiates the differentiation of osteoprogenitor cells and may facilitate the formation of bone.