Apatite nanosheets inhibit initial smooth muscle cell proliferation by damaging cell membrane.

Understanding how nanostructured coatings interact with cells is related to how they manipulate cell behaviors and is therefore critical for designing better biomaterials. The apatite nanosheets were deposited on metallic substrates via biomimetic precipitation. Cell viability of apatite nanosheets towards to smooth muscle cells (SMCs) were investigated, and the underlying mechanism was proposed. Apatite nanosheets presented inhibitory activity on SMC growth, and caused rupture of cell membranes. On the basis of measuring changes in intracellular calcium ([Ca2+]i), observing cell contraction and apatite nanosheets - SMC interaction, it was found that calcium ions released from apatite led to rises in [Ca2+]i, which induced vigorous SMC contraction on apatite nanosheets. Consequently, the cell membrane of individual SMCs was cut/penetrated by the sharp edges of apatite nanosheets, resulting in cell inactivation. This damage of cell membranes suggests a novel mechanism to manipulate cell viability, and may offer insights for the better design of calcium-based nanostructured coatings or other biomedical applications.

Bone regeneration in ceramic scaffolds with variable concentrations of PDRN and rhBMP-2.

This study evaluated the bone regeneration capacity and mechanical properties of block-type hydroxyapatite (HA)/tricalcium phosphate (TCP) scaffolds in response to different concentrations of polydeoxyribonucleotide (PDRN) and recombinant human bone morphogenic protein 2 (rhBMP-2). Thirty-two male white rabbits were used as a model of calvarial bone defect and classified into eight groups according to type and concentration of growth factor administered, viz., control group (only HA/TCP scaffold), scaffold + PDRN (0.1, 1, 5, and 10 mg/mL each) and scaffold + rhBMP-2 (0.01, 0.05, and 0.1 mg/mL each). The specimens were evaluated using histomorphometric and radiological analyses. Histomorphometric analyses indicated that the administration of PDRN did not increase bone formation. However, significant increases in bone formation were observed with the administration of rhBMP-2 at 0.05 and 0.10 mg/mL on week 8 compared to the control (p < 0.05). Radiological analyses revealed a significant increase in bone formation at week 8 with the administration of PDRN at 5 mg/mL and 10 mg/mL, and rhBMP-2 at 0.05 or 0.10 mg/mL compared to the control (p < 0.05). Our findings show that block-type HA/TCP scaffolds possess sufficient mechanical strength and bone regeneration capacity when used with optimal concentrations of growth factors.

Lateral Window Design for Maxillary Sinus Graft Based on the Implant Position.

The purpose of this study was to devise a classification and lateral window design method based on implants and to evaluate whether these classifications and methods are applicable to clinical practice. When applying the maxillary sinus elevation technique using the lateral window, possible situations were classified into four: (A) two or more sites for implants are required for maxillary sinus augmentation, (B) a single implant is required when there are no adjacent teeth, (C) a single implant is required when one adjacent tooth is present at the mesial or distal area, and (D) a single implant is required when both mesial and distal adjacent teeth are present. In order to verify whether this classification can be used in all situations, 76 patients who underwent maxillary sinus elevation with a lateral window were selected and investigated. Of them, 47 (62%) were included in Group A, 9 (12%) in Group B, 8 (11%) in Group C, and 12 (15%) in Group D. Lateral window designing in the lateral approach of sinus augmentation can be classified into four clinical situations. There were no unclassified cases. This classification and window positioning method can be applied to most cases.

3D-Printed Ceramic Bone Scaffolds with Variable Pore Architectures.

This study evaluated the mechanical properties and bone regeneration ability of 3D-printed pure hydroxyapatite (HA)/tricalcium phosphate (TCP) pure ceramic scaffolds with variable pore architectures. A digital light processing (DLP) 3D printer was used to construct block-type scaffolds containing only HA and TCP after the polymer binder was completely removed by heat treatment. The compressive strength and porosity of the blocks with various structures were measured; scaffolds with different pore sizes were implanted in rabbit calvarial models. The animals were observed for eight weeks, and six animals were euthanized in the fourth and eighth weeks. Then, the specimens were evaluated using radiological and histological analyses. Larger scaffold pore sizes resulted in enhanced bone formation after four weeks (p < 0.05). However, in the eighth week, a correlation between pore size and bone formation was not observed (p > 0.05). The findings showed that various pore architectures of HA/TCP scaffolds can be achieved using DLP 3D printing, which can be a valuable tool for optimizing bone-scaffold properties for specific clinical treatments. As the pore size only influenced bone regeneration in the initial stage, further studies are required for pore-size optimization to balance the initial bone regeneration and mechanical strength of the scaffold.

Bone Regeneration Capability of 3D Printed Ceramic Scaffolds.

In this study, we evaluated the bone regenerative capability of a customizable hydroxyapatite (HA) and tricalcium phosphate (TCP) scaffold using a digital light processing (DLP)-type 3D printing system. Twelve healthy adult male beagle dogs were the study subjects. A total of 48 defects were created, with two defects on each side of the mandible in all the dogs. The defect sites in the negative control group (sixteen defects) were left untreated (the NS group), whereas those in the positive control group (sixteen defects) were filled with a particle-type substitute (the PS group). The defect sites in the experimental groups (sixteen defects) were filled with a 3D printed substitute (the 3DS group). Six dogs each were exterminated after healing periods of 4 and 8 weeks. Radiological and histomorphometrical evaluations were then performed. None of the groups showed any specific problems. In radiological evaluation, there was a significant difference in the amount of new bone formation after 4 weeks (p < 0.05) between the PS and 3DS groups. For both of the evaluations, the difference in the total amount of bone after 8 weeks was statistically significant (p < 0.05). There was no statistically significant difference in new bone between the PS and 3DS groups in both evaluations after 8 weeks (p > 0.05). The proposed HA/TCP scaffold without polymers, obtained using the DLP-type 3D printing system, can be applied for bone regeneration. The 3D printing of a HA/TCP scaffold without polymers can be used for fabricating customized bone grafting substitutes.

Osteogenesis of 3D printed macro-pore size biphasic calcium phosphate scaffold in rabbit calvaria.

To investigate the osteogenesis of macro-pore sized bone scaffolds, biphasic calcium phosphate scaffolds with accurately controlled macro-pore size (0.8, 1.2, and 1.6 mm) and identical porosity of 70% were fabricated by the 3D printing technology. Eight New Zealand rabbits were selected in the present study, while four 8-mm-diameter calvarial defects were created in each rabbit to place BCP scaffolds with different macro-pore size. The harvested specimens of four and eight weeks were used to evaluate the bone forming ability by micro CT and histological examination. All 3D-printed BCP scaffolds exhibited excellent mechanical properties and had better bone-forming ability than the control at both four and eight weeks. Among them, scaffold with 0.8 mm pore size was superior for initial bone formation and maturation, resulting in the highest value of total bone formation.

Biomimetic apatite formed on cobalt-chromium alloy: A polymer-free carrier for drug eluting stent.

In this study, sirolimus (SRL) was loaded within biomimetic apatite formed on cobalt-chromium (Co-Cr) alloy, which has been reported for the first time, to inhibit the in-stent restenosis. Two different groups of loading SRL within biomimetic apatite were prepared: Group A (mono-layer of apatite/SRL) and Group B (bi-layer of apatite/SRL). Group A and Group B showed the biphasic pattern of SRL release up to 40 and 90days, respectively. The attachment of human artery smooth muscle cell (HASMC) for both Group A and Group B was significantly inhibited, and proliferation dramatically decreased with the release of SRL. Noteworthily, biomimetic apatite alone also suppressed the SMC proliferation. The porous biomimetic apatite uniformly covered Co-Cr stent without crack or webbings. After balloon expansion, the integrity of biomimetic apatite was sufficient to resist delamination or destruction. Thus, this study demonstrated that biomimetic apatite is a promising drug carrier for potential use in stents.

Silk fibroin membrane used for guided bone tissue regeneration.

With the aim to develop a novel membrane with an appropriate mechanical property and degradation rate for guided bone tissue regeneration, lyophilized and densified silk fibroin membrane was fabricated and its mechanical behavior as well as biodegradation property were investigated. The osteoconductive potency of the silk fibroin membranes were evaluated in a defect rabbit calvarial model. Silk fibroin membrane showed the modulated biodegradable and mechanical properties via ethanol treatment with different concentration. The membrane could prevent soft tissue invasion from normal tissue healing, and the amounts of new bone and defect closure with silk fibroin membrane were similar to those of commercially available collagen membrane.

Evaluation of Different Combinations of Biphasic Calcium Phosphate and Growth Factors for Bone Formation in Calvarial Defects in a Rabbit Model.

The aim of the present preclinical study was to investigate the capability of a new formulation of biphasic calcium phosphate (BCP) in achieving new bone formation either by itself or in combination with different concentrations of growth factors. Twenty-four 3-month-old male New Zealand white rabbits (weight range, 2.5 to 3.0 kg) that had been bred exclusively for biomedical research purposes and obtained from a licensed vendor were used. Four calvarial defects were created in each animal, for a total of 96 defects. Each defect received alloplastic BCP (Osteon III, Genoss) that was composed of 60% hydroxyapatite and 40% ß-tricalcium phosphate) (porosity, ~80%; macropore size, 200 to 400 µm; crystallinity, 95%) combined with different concentrations of recombinant human platelet-derived growth factor BB (rhPDGF-BB), human recombinant basic fibroblast growth factor-2 (rhFGF-2), or recombinant human bone morphogenetic protein-2 (rhBMP-2). A custom-made polycarbonate tube was fixed to each defect site by applying slight pressure, and a mixture of bone graft and growth factor was implanted into the tubes. Data were collected 2, 4, and 8 weeks after creation of the defects to assess early and late healing. Various amounts of newly formed bone and remnant BCP particles formed inside of the tube throughout the study period. The BCP + 0.5 mg/mL rhBMP-2 group exhibited the most bone formation. At 8 weeks, more new bone formation was noted in the Osteon III + rhBMP-2 combined group than in other groups. The present study results indicate that BCP can be combined with different concentrations of rhBMP-2, rhFGF-2, and rhPDGF-BB to produce new bone formation within a polycarbonate tube in calvarial defects in a rabbit model.

A 10-year retrospective radiographic study of implantium dental implants.

A retrospective radiographic study was conducted from five private dental offices on patients requesting dental implant therapy to replace their missing teeth. All implants were placed and restored by early 2000, and patients were encouraged to continue their dental care at the same office. The records were reviewed and analyzed by the clinicians, who had more than 15 years of dental experience at the time of the initial patient treatment. A total of 74 patients with 242 implants were recalled up to 10 years (mean follow-up: 9.21 ± 1.7 years) after loading. There were five implant failures from this radiographic observation period, resulting in a 97.9% dental implant survival rate. The mean crestal bone level change on the mesial aspect was -0.36 ± 1.05 mm, while the mean crestal bone level change on the distal aspect was -0.18 ± 0.96 mm. Thus, the overall mean bone loss was -0.28 ± 0.05 mm. The dental implants, which had a sandblasted, large-grit, acid-etched surface, appeared to achieve successful osseointegration in this long-term observation period, and the implant system's unique design and surface features resulted in a stable osseous crest without bone loss to the first thread.

Comparative study of Chinese hamster ovary cell versus Escherichia coli-derived bone morphogenetic protein-2 using the critical-size supraalveolar peri-implant defect model.

BACKGROUND: Chinese hamster ovary (CHO) cell-derived recombinant human bone morphogenetic protein-2 (rhBMP-2) has been introduced for spine, long bone, and craniofacial indications. Escherichia coli- (E. coli) derived rhBMP-2 displays comparable efficacy to CHO cell-derived rhBMP-2 in vitro and in small-animal models. The objective of this study is to evaluate the efficacy of E. coli-derived rhBMP-2 compared to the benchmark CHO cell-derived rhBMP-2 using an established large-animal model. METHODS: Contralateral, critical-size supraalveolar peri-implant defects in six adult male Hound Labrador mongrel dogs received CHO cell- or E. coli-derived rhBMP-2 (0.2 mg/mL) in an absorbable collagen sponge (ACS) carrier. In each quadrant, three dental implants were placed. A titanium mesh device was used to support space provision. The animals received fluorescent bone markers for qualitative evaluations. Animals were euthanized at 8 weeks for histopathologic and histometric evaluation. RESULTS: Clinical healing included significant swelling, but none of the animals experienced wound dehiscences. CHO cell- and E. coli-derived rhBMP-2 supported comparable bone formation (new bone area, 35.8 ± 3.6 versus 30.1 ± 2.2 mm(2); bone density, 31.8% ± 1.6% versus 35.6% ± 2.5%; and osseointegration, 32.9% ± 7.4% versus 33.7% ± 8.1%) without statistically significant differences between treatments. Newly formed immature delicate trabecular bone in fibrovascular marrow filled the space underneath the titanium mesh and extended coronally above the mesh. Seroma formation was frequently observed. There were no discernable qualitative histologic differences between treatments. CONCLUSION: CHO cell- and E. coli-derived rhBMP-2 in an ACS carrier appear equally effective at inducing local bone formation in support of dental implant osseointegration.

Comparison of implant stability after different implant surface treatments in dog bone.

OBJECTIVES: The purpose of this investigation was to evaluate the effects of different implant surface treatments on implant stability in dog mandibles. MATERIAL AND METHODS: A total of 30 implants (Dentium Co, Seoul, Korea) were placed in 5 dog mandibles. Bone quality was assessed at each site. Implant stability was evaluated using 2 different methods. An Osstell resonance frequency analyzer (RFA) was used to determine the stability at baseline (day 1), and 3, 6 and 10 weeks after surgery. Animals were euthanized 10 weeks after implant installation. Specimens were obtained and submitted to the laboratory processing. Sections were stained with hematoxylin and eosin for histologic and histomorphometric analyses. All implantation sites in dog mandibles demonstrated bone types II and III. RESULTS AND CONCLUSIONS: All implants showed good primary stability at baseline in terms of insertion torque. The results of this study suggest that surface treatment may have significant effects on biological stability 3 weeks after implant placement. Further studies are needed to confirm these initial observations in poor quality bone.

Enhanced bone forming ability of SLA-treated Ti coated with a calcium phosphate thin film formed by e-beam evaporation.

With an electron-beam evaporation process, a calcium phosphate (Ca-P) thin film of approximately 500 nm thick was deposited on sand blasted with large grits and acid etched (SLA) Ti without changing the typical morphology of the SLA surface. Dissolution behavior was investigated by measuring the amount of dissolved phosphate ions with ion chromatography after immersing the SLA Ti sample coated with a Ca-P film in 1 ml de-ionized water maintained at 37 degrees C for different periods of soaking time, and the surface morphology was observed with field emission scanning electron microscopy. The amount of phosphate ions increased quickly right after immersion but began to decrease after 2 days of immersion by redeposition with Ca ions as apatite, and the amount of biomimetic apatite increased with the extended soaking time. The Saos-2 cell was more attached on the coated surface, and the in vivo evaluation was that the Ca-P deposited SLA implant greatly improved the new bone formation ability.

Comparison of implant stability after different implant surface treatments in dog bone

OBJECTIVES: The purpose of this investigation was to evaluate the effects of different implant surface treatments on implant stability in dog mandibles. MATERIAL AND METHODS: A total of 30 implants (Dentium Co, Seoul, Korea) were placed in 5 dog mandibles. Bone quality was assessed at each site. Implant stability was evaluated using 2 different methods. An OsstellTM resonance frequency analyzer (RFA) was used to determine the stability at baseline (day 1), and 3, 6 and 10 weeks after surgery. Animals were euthanized 10 weeks after implant installation. Specimens were obtained and submitted to the laboratory processing. Sections were stained with hematoxylin and eosin for histologic and histomorphometric analyses. All implantation sites in dog mandibles demonstrated bone types II and III. RESULTS AND CONCLUSIONS: All implants showed good primary stability at baseline in terms of insertion torque. The results of this study suggest that surface treatment may have significant effects on biological stability 3 weeks after implant placement. Further studies are needed to confirm these initial observations in poor quality bone.

Effects of calcium phosphate coating to SLA surface implants by the ion-beam-assisted deposition method on self-contained coronal defect healing in dogs.

The aim of this study was to evaluate the healing of self-contained coronal defects on a sand-blasted, large-grit, acid-etched (SLA) surface implant, which had a calcium phosphate (CaP) coating applied by ion-beam-assisted deposition (IBAD). We also evaluated the effect of heating the coating to different temperatures. The CaP-coated SLA implants exhibited a slightly larger bone healing capacity in the self-contained coronal defect than SLA implants, indicating that combining SLA surface implants and a CaP coating by the IBAD method had synergistic effects on bone healing. There was no difference in the healing capacity between 350 degrees C and 450 degrees C heat treatment of the coating layer.

Characterizations of the TiO(2-x) films synthesized by e-beam evaporation for endovascular applications.

Different chemical states of titanium oxide films were deposited on commercially pure Ti (CP Ti) by electron-beam evaporation at different oxygen flow rates to examine a possibility of their applications to endovascular stents. The surface morphology, chemical composition and crystal structure of the obtained titanium oxide films were analyzed by FE-SEM, XPS and XRD, respectively. As a function of the deposition parameters employed, the obtained titanium oxide films demonstrated different mixtures of anatase phase, Ti2O3 and TiO. By the formation of titanium oxide film on the CP Ti plate, the contact angle was decreased and the cellular activity of porcine aortic smooth muscle cells was increased. Post-deposition annealing was also found to be an important factor to achieve advantageous biocompatibility. When haemocompatibility was investigated by observing adhesion of blood platelets from platelet-rich plasma, less platelet adhesion was observed on titanium oxide films. These results indicated that titanium oxide film synthesized by e-beam evaporation could be applicable to coronary stents.

Evoked electromyographic technique for quantitative assessment of the innervation status of laryngeal muscles.

OBJECTIVES: The purpose of this study was to develop a minimally invasive, noninjurious evoked electromyographic technique that could accurately quantitate the level of innervation of laryngeal muscles with recurrent laryngeal nerve stimulation. METHODS: A four-phase study was conducted in 24 canines, including 1) identification of the best stimulation-recording configuration, 2) statistical analysis of sensitivity and accuracy, 3) evaluation of safety, and 4) identification of the laryngeal muscle(s) that contribute to the evoked response. RESULTS: The results demonstrated that an entirely noninvasive technique is not feasible. The stimulating cathode must be invasive to ensure discrete activation of the recurrent laryngeal nerve, whereas both recording electrodes should remain on the surface with one overlying the thyroid ala. This configuration proved to be highly accurate, with an error rate of only 6% to 7%, and with sensitivity sufficient to detect a signal in a nerve with fewer than 1% of the axons intact. There was no evidence of nerve injury in any animal over the course of 350 stimulus needle penetrations. By use of neuromuscular blockade to identify those muscles generating the surface response, the thyroarytenoid muscle was found to be the primary contributor, whereas the posterior cricoarytenoid muscle was uninvolved. CONCLUSIONS: This evoked electromyographic technique could provide quantitative information regarding the extent of muscle innervation during denervation and regeneration in case of laryngeal paralysis.

Plasma cell granuloma in cyclosporine-induced gingival overgrowth: a report of two cases with immunohistochemical positivity of interleukin-6 and phospholipase C-gamma1.

We report two cases of gingival plasma cell granuloma in a 34-yr-old and 40-yr-old two male renal transplant recipients with cyclosporine A (CsA)-induced gingival overgrowth (GO). Histologically, these lesions were composed of mature plasma cells, showing polyclonality for both lambda and kappa light chains and fibrovascular connective tissue stroma. In addition to the fact that CsA-induced plasma cell granuloma is rare, the salient features of our cases were the secretion of interleukin-6 and overexpression of phospholipase C- gamma 1 of the tumor cells, which may explain the mechanisms of CsA- induced GO.