Decontamination methods using a dental water jet and dental floss for microthreaded implant fixtures in regenerative periimplantitis treatment.

OBJECTIVE: This study evaluated decontamination methods using a dental water jet and dental floss on microthreaded implants for regenerative periimplantitis therapy. MATERIALS AND METHODS: In 6 beagle dogs, experimental periimplantitis was induced, and decontamination procedures, including manual saline irrigation (control group), saline irrigation using a dental water jet (group 1) and saline irrigation using a dental water jet with dental flossing (group 2), were performed. After in situ decontamination procedures, some of the implant fixtures (n = 4 per group) were retrieved for analysis by SEM, whereas other fixtures (n = 4 per group) underwent regenerative therapy. After 3 months of healing, the animals were killed. RESULTS: The SEM examination indicated that decontamination of the implant surfaces was the most effective in group 2, with no changes in implant surface morphology. The histological examination also revealed that group 2 achieved significantly greater amounts of newly formed bone (6.75 ± 2.19 mm; P = 0.018), reosseointegration (1.88 ± 1.79 mm; P = 0.038), and vertical bone fill (26.69 ± 18.42%; P = 0.039). CONCLUSION: Decontamination using a dental water jet and dental floss on microthreaded implants showed positive mechanical debridement effects and positive bone regeneration effects.

Determination of osteogenic or adipogenic lineages in muscle-derived stem cells (MDSCs) by a collagen-binding peptide (CBP) derived from bone sialoprotein (BSP).

Bone sialoprotein (BSP) is a mineralized, tissue-specific, non-collagenous protein that is normally expressed only in mineralized tissues such as bone, dentin, cementum, and calcified cartilage, and at sites of new mineral formation. The binding of BSP to collagen is thought to be important for initiating bone mineralization and bone cell adhesion to the mineralized matrix. Several recent studies have isolated stem cells from muscle tissue, but their functional properties are still unclear. In this study, we examined the effects of a synthetic collagen-binding peptide (CBP) on the differentiation efficiency of muscle-derived stem cells (MDSCs). The CBP sequence (NGVFKYRPRYYLYKHAYFYPHLKRFPVQ) corresponds to residues 35-62 of bone sialoprotein (BSP), which are located within the collagen-binding domain in BSP. Interestingly, this synthetic CBP inhibited adipogenic differentiation but increased osteogenic differentiation in MDSCs. The CBP also induced expression of osteoblastic marker proteins, including alkaline phosphatase (ALP), type I collagen, Runt-related transcription factor 2 (Runx2), and osteocalcin; prevented adipogenic differentiation in MDSCs; and down-regulated adipose-specific mRNAs, such as adipocyte protein 2 (aP2) and peroxisome proliferator-activated receptor γ. The CBP increased Extracellular signal-regulated kinases (ERK) 1/2 protein phosphorylation, which is important in lineage determination. These observations suggest that this CBP determines the osteogenic or adipogenic lineage in MDSCs by activating ERK1/2. Taken together, a novel CBP could be a useful candidate for regenerating bone and treating osteoporosis, which result from an imbalance in osteogenesis and adipogenesis differentiation.

Ex vivo bone morphogenetic protein-2 gene delivery using gingival fibroblasts promotes bone regeneration in rats.

AIM: The aim of the present study was to investigate bone regeneration following ex vivo bone morphogenetic protein-2 (BMP-2) gene delivery using human gingival fibroblasts (HGFs) in rat calvarial defects. MATERIALS AND METHODS: An 8 mm craniotomy defect was created in Sprague-Dawley rats. The animals were divided into four groups: (1) non-grafted group, the defect was left empty; (2) collagen matrix group, the defect was filled with collagen matrix only; (3) HGF group, the defect was filled with non-transduced HGFs on collagen matrix; (4) BMP-2/HGF group, the defect was filled with BMP-2 gene-transduced HGFs on collagen matrix. Animals were sacrificed at 2 and 4 weeks after surgery, and micro-computed tomographic and histologic observations were performed. RESULTS: The BMP-2/HGF group showed promoted osseous healing of calvarial defects, as compared with the other groups. At both 2 and 4 weeks, regenerated bone area was significantly greater in the BMP-2/HGF group than the other three groups. Quite a few number of transplanted HGFs were observed within the regenerated bone tissues. CONCLUSIONS: The results of this study suggest that ex vivo BMP-2 gene delivery induces prominent bone regeneration in vivo and HGFs may be useful as target cells for ex vivo gene therapy.

Bone regeneration into side openings and hollow inner channel of a dental implant.

The current study examined whether bone can regenerate into an open space fabricated inside the metal implant and maintain its quantity and quality at the early post-implantation healing periods. 12 conventional one piece screw type titanium dental implants (control group) and 12 hybrid dental implants with spiral side openings (0.58 mm wide) connected to hollow inner channel (experimental group) were bilaterally placed in each quadrant at the P3, P4 and M1 positions in mandible of 4 adult beagles following 2 months of post-extraction healing. Fluorescent bone labels to qualitatively evaluate newly formed bone tissues were administered at 2 and 4 weeks of post-implantation periods, respectively. 3 control and 3 experimental bone-implant constructs for each animal were dissected from 2 animals at each 3 and 6 weeks of post-implantation healing periods. Undecalcified specimens were prepared from each construct for histological analyses to measure bone-to-implant contact (BIC) and interfacial bone area (BA), and also for nanoindentation and scanning electron microscopy to assess elastic modulus (E) and composition of bone tissues surrounding the implants, respectively. A substantial amount of newly formed bone tissues were observed at the implant interfaces of both implant groups. Bone tissues successfully regenerate through the side openings and hollow inner channel of the experimental implant as early as 3 weeks of post-implantation healing. The E values of the newly formed bone tissues were measured comparable to those of normal bone tissues. The current results indicate that the new hybrid implant can conduct bone regeneration into the inner architecture, which likely improves stability of the implant system by enhancing integrity of implant with interfacial bone.

Identification of cell-penetrating osteogenic peptide from copine-7 protein and its delivery system for enhanced bone formation.

Peptide and proteins are recognized as highly selective and therapeutically active biomaterials, as well as relatively safe in clinical application. A calcium phospholipid-binding protein, copine 7 (CPNE7), has been recently identified to induce hard tissue regeneration, including bone and dentin by internalizing into the cells. However, the clinical application of the full length of CPNE7 has limited due to its large size with short half-life. Herein, as an alternative to CPNE7, six bioactive synthetic peptides are designed from CPNE7 (CPNE7-derived peptides, CDP1-CDP6) and investigated their osteogenic potential. Among the CDPs, CDP4 have the highest level of cell-penetrating activity as well as osteogenic efficiency in dental pulp stem cells (DPSCs). CDP4 increased the expression of osteogenesis-related genes and proteins, which was comparable to that by BMP-2. The cell penetration capacity of CDP4 may synergistically induce the osteogenic potential of DPSCs. Moreover, the implantation of the mixture of CDP4 with injectable collagen gel increased bone formation with recovery in the mouse calvarial defect model, comparable to full-length CPNE7 and even BMP-2. In conclusion, these results suggest that our synthetic peptide, CDP4, can be applied in combination with biomaterial to provide high osteogenic efficacy in the field of bone tissue engineering.

Effect of scaling and root planing on alveolar bone as measured by subtraction radiography.

BACKGROUND: Scaling and root planing of diseased periodontal pockets is fundamental to the treatment of periodontal disease. Although various clinical parameters have been used to assess the efficacy of this therapy, radiographic analysis of changes in bone density following scaling and root planing has not been extensively researched. In this study, digital subtraction radiography was used to analyze changes that occurred in the periodontal hard tissues following scaling and root planing. METHODS: Thirteen subjects with a total of 39 sites that presented with >3 mm of vertical bone loss were included in this study. Clinical examinations were performed and radiographs were taken prior to treatment and were repeated 6 months following scaling and root planing. Radiographic analysis was performed with computer-assisted radiographic evaluation software. Three regions of interest (ROI) were defined as the most coronal, middle, and apical portions of each defect. A fourth ROI was used for each site as a control region and was placed at a distant, untreated area. Statistical analysis was carried out to evaluate changes in the mean gray level at the coronal, middle, and apical region of each treated defect. RESULTS: Digital subtraction radiography revealed an increase in radiographic density in 101 of the 117 test regions (83.3%). A 256 gray level was used, and a value >128 was assumed to represent a density gain in the ROI. The average gray level increase was 18.65. Although the coronal, middle, and apical regions displayed increases in bone density throughout this study, the bone density of the apical ROI (gray level = 151.27 +/- 20.62) increased significantly more than the bone density of the coronal ROI (gray level = 139.19 +/- 21.78). A significant increase in bone density was seen in probing depths >5 mm compared to those <5 mm in depth. No significant difference was found with regard to bone-density changes surrounding single- versus multiple-rooted teeth. CONCLUSION: Scaling and root planing of diseased periodontal pockets can significantly increase radiographic alveolar bone density as demonstrated through the use of digital subtraction radiography.

Assembly of collagen-binding peptide with collagen as a bioactive scaffold for osteogenesis in vitro and in vivo.

Bioactive scaffolds inducing cell adhesion, differentiation have been premise for optimal formation of target tissue. Collagen has been employed as a tissue regenerative scaffold especially for bone regeneration and has been chemically surface-modified to present bioactivity. Herein, we show that peptide, denoted as collagen-binding motif (CBM, GLRSKSKKFRRPDIQYPDATDEDITSHM) identified from osteopontin (OPN) protein, was able to specifically bind collagen without chemical conjugation, while presenting apatite forming capability in vitro and in vivo. Collagen surface alone was not able to induce noticeable apatite nucleation however, mineralization was evident when assembled with CBM peptide, implying that the collagen-CBM assembly played a pivotal role in biomineralization. In vivo result further demonstrated that the CBM peptide in complex with material was able to induce bone formation by helping mineralization in the bone defect. Taken together, the CBM peptide herein and its assembly with collagen can be applied as an inducer of biomineralization as well as a bioactive scaffold for bone regeneration.

Enhanced bone regeneration in beagle dogs with bovine bone mineral coated with a synthetic oligopeptide.

BACKGROUND: Recombinant human bone morphogenic protein-2 stimulates bone augmentation in animal models. The aim of this study was to evaluate the capacity of bovine bone mineral coated with synthetic oligopeptides to enhance guided bone regeneration in the beagle 3-wall defect model and the clinical implications. METHODS: The second and fourth mandibular premolars of four adult beagle dogs were extracted bilaterally, and the extraction sites were allowed to heal for 2 months. An L-shaped defect was prepared at the central part of the extraction site with a round bur on a low-speed motor. Peptide-coated bone mineral was implanted on one side, and uncoated bone mineral was implanted on the other side. The membrane was tucked underneath the mobilized lingual flap. New bone formation at the test and control sites was determined at 4 weeks. RESULTS: No specimen revealed any evidence of infection or foreign body reaction, and all wounds showed a good healing response. Sites augmented with peptide-coated bone mineral and uncoated mineral exhibited excellent maintenance of the ridge contour. There was more new bone at sites with peptide-coated bone mineral than at control sites. The new bone in sites with peptide-coated bone mineral was deposited evenly around the graft material, and bone mineral was integrated fully into the new bone. CONCLUSION: Deproteinized cancellous bovine bone coated with synthetic oligopeptides enhanced new bone formation, and it seemed to be a better material for guided bone regeneration in the beagle L-shaped defect model.

Osteopromotion with synthetic oligopeptide-coated bovine bone mineral in vivo.

BACKGROUND: Recombinant human bone morphogenetic protein (rhBMP)-2 has been shown to stimulate significant regeneration of alveolar bone and cementum in periodontal defects clinically. The aim of this study was to evaluate the osteopromotive effect of oligopeptide domain-coated bovine bone mineral granules in a rabbit calvarial defect model. METHODS: The peptide-coated bovine bone was fabricated by incubating the graft material in a solution of oligopeptide domain. The cell attachment and proliferation were measured in vitro. Peptide-coated (test group) or uncoated (control group) bone minerals were implanted into 10-mm calvarial defects in rabbits, and the animals were sacrificed at 1, 2, or 4 weeks post-surgery. RESULTS: The cells grown with peptide-coated bone showed significantly higher proliferation activity at all times compared to cells grown with peptide-uncoated bone mineral. At 2 weeks post-surgery, the control wounds showed a limited amount of osteoid formation in a centripetal pattern around the grafted bone, but the oligopeptide domain-coated grafted bone had formed new bone around the grafted area. At 4 weeks post-surgery, the control sites showed increased new bone formation, but they still showed a significant difference from the peptide-coated bone-implanted sites. CONCLUSIONS: At 2 and 4 weeks, accelerated new bone formation was observed within the experimental sites compared to control groups. The use of deproteinized bovine bone combined with a synthetic oligopeptide seems to be a more beneficial material for bone regeneration in the early healing period.

Influence of RGD-containing oligopeptide-coated surface on bone formation in vitro and in vivo.

New bone formation on an RGD-containing oligopeptide-coated surface in vitro and in vivo was investigated. The surface showed two-fold higher osteoblastic cell adhesion and differentiation in vitro, and revealed statistically significant in vivo bone formation compared with the control (P < 0.05).

Comparative Study of rhPDGF-BB Plus Equine-Derived Bone Matrix Versus rhPDGF-BB Plus ß-TCP in the Treatment of Periodontal Defects.

The purpose of this study was to evaluate the efficacy and safety of equine-derived bone matrix as a carrier for recombinant human platelet-derived growth factor BB (rhPDGF-BB) versus beta-tricalcium phosphate (ß-TCP) for the treatment of intraosseous periodontal defects in adult patients. This study was performed on 32 adults with advanced periodontal disease. Eligible subjects were randomized in 1:1 ratio into a test (rhPDGF-BB-coated equine-derived bone matrix) or control group (rhPDGF-BB-coated ß-TCP). Probing pocket depth (PD), clinical attachment level (CAL), gingival recession (GR), and defect depth on radiographs were measured at 2 weeks before surgery, on the day of surgery (DOS), and 6 months postsurgery (6MPS). The clinical and radiographic data were analyzed over the test period. Statistically significant PD reductions and CAL gain between baseline and 6MPS and between ODS and 6MPS were seen in both groups (P < .01). No statistically significant differences in PD reduction were found between groups. However, the test group showed significant CAL gain between DOS and 6MPS. The radiographic bone level change was statistically significant compared to baseline (P < .01) in both groups. The results suggested that equine-derived bone matrix is a viable, effective, and safe carrier scaffold for rhPDGF in periodontal defects.

Enhanced osteogenic promotion around dental implants with synthetic binding motif mimicking bone morphogenetic protein (BMP)-2.

Synthetic receptor binding motif mimicking bone morphogenetic protein-2 (BMP-2) was covalently linked to titanium (Ti) surfaces through a chemical conjugation process. The composition and properties of surface-modified Ti were investigated by XPS as well as by measuring surface radioactivity. In vitro tests were conducted with osteoblast-like MC3T3-E1 cells to assess cell attachment, morphology, and expression of osteogenic marker in the cells grown on modified Ti surfaces. In addition, in vivo experiments involved implants in mandibular bone defects of beagles to evaluate the effect of surface modification on bone regeneration. Results of XPS measurements showed a complete and homogeneous peptide overlayer on the Ti surfaces; the content was further measured by gamma counting. Biological evaluations showed that the biochemically modified Ti samples were active in terms of cell attachment behavior. The MC3T3-E1 cell growth rate, marker protein expression, and alkaline phosphatase production of the peptide-modified surfaces were all higher than those of control Ti. Importantly, the implants in the canine mandibles showed significant increase of bone growth when modified with bioactive peptide, thereby confirming that biochemical modifications of Ti surfaces can enhance the rate of bone healing as compared with untreated Ti surfaces.

Multiplex polymerase chain reaction detection of black-pigmented bacteria in infections of endodontic origin.

The purpose of this study was to detect the presence of Porphyromonas endodontalis, P. gingivalis, Prevotella intermedia, P. nigrescens, and P. tannerae from clinical samples using multiplex polymerase chain reactions (PCR). Two different multiplex PCR protocols were used (one for the two Porphyromonas species and the other for the three Prevotella species), each one using a primer pair specific for each target species. The results were compared to those of the conventional culture procedures. Microbial samples were taken aseptically from 40 infected root canals and abscesses from patients. Samples were cultured in an anaerobic condition for conventional identification using a Rapid ID 32 A kit. Multiplex PCR was processed using the DNA extracted from each sample. At least one of the five species of black-pigmented bacteria (BPB) were detected in 65% (26 of 40) of the samples using multiplex PCR, and in 15% (6 of 40) using the conventional culture procedures. Multiplex PCR was more rapid, sensitive, specific, and effective in detecting BPB than the conventional culture procedures.

The effect of the surface modification of titanium using a recombinant fragment of fibronectin and vitronectin on cell behavior.

The surface of titanium implants is in direct contact with host tissue and plays a critical role in determining biocompatibility. Fibronectin (FN) and vitronectin (VN) are major cell adhesive proteins found in the extracellular matrix (ECM) of various tissues, and in circulating blood. The aim of this study was to evaluate the engineered biomimetic surface of titanium by using recombinant fragment of FN(8-10) and VN(NTD) that contains the binding site for integrins. MC3T3-E1 cells seeded upon the FN(8-10)-coated titanium showed a marked increase in cell adhesion, proliferation, and differentiation over VN(NTD)-coated titanium. In addition, we confirmed that the surface properties of titanium prefer for FN(8-10) over VN(NTD) (p<0.05) in protein adhesion. These results suggest that the FN(8-10)-modified titanium surface can be used to improve the osseointegration of titanium implants by enhancing bone formation.

Biological efficacy of silk fibroin nanofiber membranes for guided bone regeneration.

The favorable biological properties of silk fibroin (SF) nanofiber membrane make it a good candidate for clinical applications as a device in bone and periodontal regenerative therapy. The purpose of this study is to evaluate the biocompatibility of the SF nanofiber membrane, and to examine its effect on bone regeneration in a rabbit calvarial model. To examine the biocompatibility of the electrospun SF membrane, we investigated cell proliferation, morphology, and differentiation. The bone regenerative efficacy of the membrane was evaluated in the calvarial defect of rabbits. The cell numbers and osteocalcin production labels were significantly increased in accordance with culture period. Cells had a stellate shape and broad cytoplasmic extensions on the membrane. The cells showed activity of ALPase that was comparable to culture dishes, and were calcified similarly to culture dishes. In in vivo tests, a complete bony union across the defects was observed after 8 weeks. At 12 weeks, the defect had completely healed with new bone. In conclusion, the SF nanofiber membrane was shown to possess good biocompatibility with enhanced bone regeneration and no evidence of any inflammatory reaction. These results strongly suggest that the SF membrane should be useful as a tool for guided bone regeneration.

Effects of adhesion molecules on the behavior of osteoblast-like cells and normal human fibroblasts on different titanium surfaces.

This study examined the influences of titanium (Ti) discs with similar surface roughnesses (R(a) values), but with different topographies and chemical compositions, on the adhesion, spreading, and the alkaline phosphatase (ALP) activity of osteoblast-like cells and normal human fibroblasts. The presence of adhesion molecules on the Ti surfaces and their effects on cell activity were also investigated. Two types of Ti discs were prepared. One kind was a mechanically polished Ti disc, and the other type was a disc obtained by the heating of hydroxyapatite (HA) dip-coated Ti. Scanning electron microscopy, optical interferometry, and scanning Auger electron spectroscopy were used to examine the surface morphology, roughness, and chemical composition, respectively, of the superficial Ti layer. The two types of Ti discs had different topographies and chemical compositions, but had similar R(a) values. The cells on both surface types had similar behaviors and ALP activities. A biological evaluation of the surface-modified Ti discs showed that the type I collagen coating was functionally active in terms of cell spreading in both types of Ti discs. In the mechanically polished Ti discs, fibronectin was functionally active in the normal human fibroblasts, but not in the osteoblast-like cells. Cell adhesion was slightly better on the heat-treated HA dip-coated Ti discs, but not on the mechanically polished Ti discs. Type I collagen and fibronectin mediated the adhesion and spreading of osteoblast-like cells through alpha2beta1 integrin and alpha5beta1 integrin, respectively. These results suggest that type I collagen might be a good candidate for the biochemical modification of Ti surfaces, particularly those surfaces obtained by heating of HA dip-coated Ti.

Biological evaluation of chitosan nanofiber membrane for guided bone regeneration.

BACKGROUND: Chitosan is known as a biodegradable and non-toxic natural polymer that enhances wound healing and bone formation. The aims of this study are to evaluate the biocompatibility of chitosan nanofiber membranes and to examine the effect of the chitosan nanofiber membranes on bone regeneration in rabbit calvarial defects. METHODS: In vitro cell proliferation tests using human osteosarcoma cell line MG63 and reverse transcription-polymerase chain reaction (RT-PCR) to evaluate the expression of alkaline phosphatase (ALP), collagen, osteocalcin (OCN), and GAPDH were done on chitosan nanofiber membranes. Chitosan nanofiber membranes were implanted in subcutaneous connective tissue and observed for 2, 4, and 6 weeks in rats. Ten-millimeter diameter round cranial defects were made in rabbits and covered by chitosan nanofiber membranes for 4 weeks. Specimens were decalcified and observed by a light microscope. RESULTS: MG63 cells proliferated for 28 days on the chitosan nanofiber membranes and expressed ALP, collagen, OCN, and GAPDH at 2 weeks. Chitosan nanofiber membranes that were grafted into rat subcutaneous tissue maintained their shape and space for bone regeneration for as long as 6 weeks. No inflammation could be seen on the membrane surface or in the surrounding tissues. In the rabbit calvarial defects, new bone filled the defects and fused to the original old bone in 4 weeks. CONCLUSIONS: The biocompatibility of the chitosan nanofiber membrane was confirmed, with enhanced bone regeneration and no evidence of an inflammatory reaction. This experiment shows that the novel biodegradable chitosan nanofiber membrane may be useful as a tool for guided bone regeneration.

Effect of subantimicrobial dose doxycycline as an effective adjunct to scaling and root planing.

BACKGROUND: This study evaluated the efficacy and safety of a subantimicrobial dose of doxycycline (SDD) in conjunction with scaling and root planing (SRP). METHODS: The study was a 9-month, double masked, randomized, placebo-controlled, parallel-group trial. A total of 41 patients with moderate chronic periodontitis who received SRP were randomly allocated to receive either a doxycycline hyclate or a placebo 2 weeks after SRP. Clinical attachment level (CAL), the probing depth (PD), gingival crevicular fluid (GCF) levels, and matrix metalloproteinase (MMP)-8 and -13 levels were measured throughout the study. The effect of SDD in conjunction with SRP on the dynamics of the periodontal microflora was also assessed using dark-field microscopic and culture analysis. Information on adverse events was collected throughout the study. RESULTS: During the treatment period, per-patient reductions in PD and CAL were demonstrated for both treatment groups, with a significantly greater reduction for the SDD group. The mean value of per-patient change in the GCF was much greater for the SDD group. Microbial analysis showed there were a general tendency for cocci, non-motile rods, and aerobes to increase with increasing treatment duration and a general decreasing tendency for spirochetes, motile rods, and anaerobes and black pigmented bacteria in both treatment groups, but no significant difference between the groups. The MMP-8 and -13 levels of the SDD group gradually reduced with time, and the mean perpatient average was significantly higher than in the placebo group. The adverse events in the SDD group were similar to those in the placebo group. CONCLUSION: This study suggests that a submicrobial dose of doxycycline as an adjunct therapy with SRP might be safe and effective in the long-term management of chronic periodontitis.

Biomimetic approach on human periodontal ligament cells using synthetic oligopeptides.

BACKGROUND: Periodontal ligament (PDL) cells, connecting root cementum with alveolar bone, are important for periodontal wound healing. In order to obtain a predictable periodontal regeneration, selective adhesion and proliferation of PDL cells are essential. The purpose of this study was to investigate the effects of synthetic peptides mimicking cell-binding domain of fibronectin (FN) on human PDL cells. METHODS: Two types of oligopeptides, Gly3-Pro-His-Ser-Arg-Asn-Gly6-Arg-Gly-Asp-Gly (G3PHSRNG6RGDG) and Gly3-His-Pro-Asn-Arg-Ser-Gly6-Arg-Gly-Asp-Gly (G3HPNRSG6RGDG), were constructed using a solid-phase peptide synthesizer. Fibronectin type III ninth to tenth domain (FN III 9-10) and Arg-Gly-Asp-Ser (RGDS) were prepared for comparison with the effects of synthetic peptides. These peptides were coated onto 96-well cell culture plates with 0.001 approximately 100 microM concentrations. Cultured human PDL cells were then applied to the peptide-coated wells at a density of 1 x 10(4)/well. After 1 hour incubation at 37 degrees C, adhered cells were fixed, stained, and examined by phase contrast microscopy for cell spreading assay. Attached PDL cells were solubilized with 2% sodium dodecyl sulfate (SDS) for the cell attachment assay by measuring absorbance at 595 nm in microplate reader. Western blot analysis was performed to determine extracellular signal-regulated kinase (ERK1/2) activity. RESULTS: Cell attachment and spreading assays revealed that G3PHSRNG6RGDG (> or = 10 microM) possesses similar adhesive behavior to FN III 9-10. G3PHSRNG6RGDG showed a comparable ERK1/2 activity when compared to FN III 9-10. CONCLUSIONS: G3PHSRNG6RGDG enhanced an attachment and spreading of human PDL cells thereby increasing ERK1/2 activity. Taken together, it is anticipated that this peptide might be a potential tool for arranging a biologically attractive environment for PDL cells, which would enhance periodontal regeneration efficacy.

Enhanced bone augmentation by controlled release of recombinant human bone morphogenetic protein-2 from bioabsorbable membranes.

BACKGROUND: The present study was undertaken to determine the effect of recombinant human bone morphogenetic protein-2 (rhBMP-2)-loaded biodegradable membranes on bone augmentation in a rabbit calvarial model. METHODS: Five microg of rhBMP-2 was loaded into a stiff hemispherical dome membrane made of poly(L-lactide) and tricalcium phosphate (PLLA/TCP). The release kinetics of rhBMP-2 from the membrane were determined in vitro using a human BMP-2 immunoassay. Twelve rhBMP-2-loaded dome membranes (test group) and 12 control dome membranes (control group) were placed on the partial-thickness calvarial defects of 24 rabbits. The animals were sacrificed at 4 and 8 weeks, and undecalcified ground sections were prepared. Newly formed bone area and height were measured histomorphometrically and calculated by percentage ratio to the total submembranous space area and height below the dome. RESULTS: In vitro release results demonstrated that rhBMP-2 was released consistently over a 4-week period following a high initial burst release on the first day. At both 4 and 8 weeks, histomorphometric analysis revealed that the test group showed significantly higher newly formed bone heights and areas than the control group (P < 0.01). In the control group, new bone height was 36.3% of the dome height and the new bone area reached 8.2% of the submembranous space area at 8 weeks, while the test group reached 87.3% and 35.4%, respectively. CONCLUSION: These results suggest that the use of rhBMP-2-loaded PLLA/TCP membranes can result in additional bone augmentation, which is due to the osteoinductive properties of rhBMP-2 released from the membrane during healing.