Bone tissue modelling and remodelling following guided bone regeneration in combination with biphasic calcium phosphate materials presenting different microporosity.

OBJECTIVES: The aim of this study was to investigate bone regeneration following application of a novel biphasic calcium phosphate (BCP I) composed of microstructured granules of 90% ß-tricalcium phosphate (ß-TCP)/10% hydroxyapatite (HA) compared to BCP non-microstructured biphasic calcium phosphate with a composite of 60% hydroxyapatite/40% ß-TCP (BCP II) and a deproteinized bovine bone mineral (DBBM) at surgically created defects in the mandible of minipigs in a combined approach with guided bone regeneration (GBR). MATERIAL AND METHODS: Sixteen minipigs were used for the study. Lower premolars P2, P3, P4 and first molar M1 were extracted. Following 3 months of healing, two defects with a width and depth of 7 mm were created bilaterally in the mandible. The different grafting materials were randomly placed in the created defects and covered by means of a collagen membrane. After 3 and 8 weeks, biopsies were sampled. All specimens were evaluated with descriptive histology and histomorphometric evaluations complemented by micro-CT scan analysis. RESULTS: All three biomaterials presented with higher bone volume at 8 weeks compared to 3 weeks (P < 0.0442). BCP I and DBBM demonstrated a significant higher amount of bone formation compared to BCP II at 8 weeks (P < 0.0328). BCP I also demonstrated a significant higher percentage of remaining graft volume compared to the other test groups both at 3 and 8 weeks (P < 0.0001 to P < 0.0003). Congruently, defects containing BCP I showed a significant higher amount of mineralized tissue compared to the other groups. CONCLUSIONS: All the three test materials performed well with regard to bone formation at 8 weeks. BCP I showed significant higher amounts of newly formed bone despite a higher remaining graft volume compared to the other groups. With regard to the regenerative outcome, all the three materials can be recommended for clinical use.

Nanostructures and hydrophilicity influence osseointegration: a biomechanical study in the rabbit tibia.

OBJECTIVE: Implant surface properties have long been identified as an important factor to promote osseointegration. The importance of nanostructures and hydrophilicity has recently been discussed. The aim of this study was to investigate how nanostructures and wettability influence osseointegration and to identify whether the wettability, the nanostructure or both in combination play the key role in improved osseointegration. MATERIALS AND METHODS: Twenty-six adult rabbits each received two Ti grade 4 discs in each tibia. Four different types of surface modifications with different wettability and nanostructures were prepared: hydrophobic without nanostructures (SLA), with nanostructures (SLAnano); hydrophilic with two different nanostructure densities (low density: pmodSLA, high density: SLActive). All four groups were intended to have similar chemistry and microroughness. The surfaces were evaluated with contact angle measurements, X-ray photoelectron spectroscopy, scanning electron microscopy, atomic force microscopy and interferometry. After 4 and 8 weeks healing time, pull-out tests were performed. RESULTS: SLA and SLAnano were hydrophobic, whereas SLActive and pmodSLA were super-hydrophilic. No nanostructures were present on the SLA surface, but the three other surface modifications clearly showed the presence of nanostructures, although more sparsely distributed on pmodSLA. The hydrophobic samples showed higher carbon contamination levels compared with the hydrophilic samples. After 4 weeks healing time, SLActive implants showed the highest pull-out values, with significantly higher pull-out force than SLA and SLAnano. After 8 weeks, the SLActive implants had the highest pull-out force, significantly higher than SLAnano and SLA. CONCLUSIONS: The strongest bone response was achieved with a combination of wettability and the presence of nanostructures (SLActive).

A novel three-dimensional analysis of standardized bone defects by means of confocal scanner and micro-computed tomography.

OBJECTIVES: The aim of the study was to introduce a novel three-dimensional (3D) method to quantify the relative amount of different tissue components in bone substitute-treated defects by means of integration of confocal laser imaging into micro-computed tomography (µCT) analysis. MATERIALS AND METHODS: One standardized semisaddle intraosseous defect was prepared in the mandibles of six minipigs and scanned by an optical scanner to capture the surface of the fresh defect in a 3D manner. Subsequently, all the defects were filled with a biphasic calcium phosphate material. The animals were divided into two groups of three animals each, which were allowed to heal for 3 and 8 weeks, respectively. µCT analysis followed the two healing periods and was performed on all defect locations. The data from optical scanning and µCT were used for three-dimensional evaluation of bone formation, nonmineralized tissue ratio, and graft degradation. The integration of confocal laser scanning into µCT analysis through a superimposition imaging procedure was conducted using the software Amira (Mercury Computer Systems, Chelmsford, MA, USA). RESULTS: The feasibility of combining the confocal imaging into µCT data with regard to obtaining accurate 3D quantification was demonstrated. The amount of tissue components was identified and quantified in all the investigated samples. Quantitative analysis demonstrated that a significant increase in the amount of bone filling the defect was observed in vivo (p < 0.02) while a significant decrease in the amount of nonmineralized tissue occurred (p < 0.04). No difference in the amount of residual grafting material was detected between 3 and 8 weeks in vivo (p > 0.38). CONCLUSIONS: The combination of confocal imaging and micro-computed tomography techniques allows for analysis of different tissue types over time in vivo. This method has revealed to be a feasible alternative to current bone regeneration quantification methods. CLINICAL RELEVANCE: Assessment of bone formation in a large animal model is a key step in assessing the performance of new bone substitute materials. Reliable and accurate methods are needed for the analysis of the regenerative potential of new materials.

A Comparison of Tapered and Nontapered Implants in the Minipig.

PURPOSE: Tapered implants are thought to result in equivalent long-term stability and marginal peri-implant bone height in comparison to cylindrical implants. The goal of this study was to compare the primary stability and osseointegration of a novel bone-level tapered-wall implant to a control bone-level cylindrical implant with exactly the same surface characteristics, in a direct side-by-side comparison in an animal model. MATERIALS AND METHODS: Mandibular premolars and first molars were extracted in 11 G.ttingen minipigs, and healing was allowed for 3 months. Six implants were placed with a 4-week surgical lag time between hemimandibles (three per side), and either immediately loaded (first implantation time point) or submerged (second implantation time point). Eight-mm-long × 4.1-mm-diameter titanium screw-type bone-level implants with tapered (two per side) and cylindrical bodies (one per side) were used (Institut Straumann); insertion torque and implant stability were measured, and the soft tissue was closed. Standardized radiographs were taken at implant placement and at sacrifice. Following a healing period of 1 month from the last surgical intervention (8 weeks total healing time for loaded implants; 4 weeks total healing time for nonloaded implants), the animals were sacrificed and mandibular blocks were harvested for nondecalcified histologic and histomorphometric analysis. RESULTS: All implants were integrated radiographically and osseointegrated histologically. Maximum insertion torque measurements and implant stability quotient values showed no significant difference between tapered and cylindrical implants. Histomorphometric analysis also resulted in comparable bone-to-implant contact values between the implant types and similarly limited marginal peri-implant bone resorption; no significant differences were observed regarding all the evaluated parameters between the groups, regardless of the loading regime. CONCLUSION: In a direct side-by-side comparison, in an intraoral minipig model, a novel bone-level tapered screw implant with an SLActive surface showed comparable clinical and histologic outcomes to a parallel-walled bone-level screw implant with an SLActive surface.

Convergent methods assessing bone growth in an experimental model at dental implants in the minipig.

Implant dentistry demonstrated its reliability in treating successfully an increased amount of patients with dental implants exhibiting hydrophilic (modSLA) or non-hydrophilic (SLA) surfaces. Objectives of this minipig study consisted in assessing bone regeneration at both types of dental implants in a new experimental model (lateral bone defects) by implementing a convergent analysis approach combining histology and contact radiography. In six adult female minipigs standardized acute alveolar defects were created, then receiving two implants in the mandibles bilaterally (modSLA and SLA, one of each combined with Guided Bone Regeneration). Animals were sacrificed after 28 days of healing. Mid-crestal specimen were analyzed assessing missing BIC to implant shoulder (fBIC); vertical bone growth upon implant surface, bone:tissue ratio and grafting material area occupied by material remnants for GBR sites. Values obtained from both analyses were compared and statistical correlations scrutinized. Although dimensions of mean differences and adjusted means given by radiographic method were lower than histological ones, comparison of different implants yielded similar results. Statistical analyses of correlation and concordance coefficients used to evaluate radiological method of measurement showed high level of concordance (concordance coefficient=0.912 and correlation coefficient=0.939) for fBIC. Similar results were observed for vertical new bone and for remnants of graft. Discrepancies for new bone and for mineralized tissue resulted in concordance coefficient of 0.182 and 0.054. The results indicate that contact X-rays can be used for morphometric assessments regarding defect fill; however, histological staining remains beneficial if greater resolution for distinguishing qualitative differences in the tissues is required.

Nonwoven-based gelatin/polycaprolactone membrane proves suitability in a preclinical assessment for treatment of soft tissue defects.

Standard preclinical assessments in vitro often have limitations regarding their transferability to human beings, mainly evoked by their nonhuman and tissue-different/nontissue-specific source. Here, we aimed at employing tissue-authentic simple and complex interactive fibroblast-epithelial cell systems and their in vivo-relevant biomarkers for preclinical in vitro assessment of nonwoven-based gelatin/polycaprolactone membranes (NBMs) for treatment of soft tissue defects. NBMs were composed of electrospun gelatin and polycaprolactone nanofiber nonwovens. Scanning electron microscopy in conjunction with actin/focal contact integrin fluorescence revealed successful adhesion and proper morphogenesis of keratinocytes and fibroblasts, along with cells' derived extracellular matrix deposits. The "feel-good factor" of cells under study on the NBM was substantiated by forming a confluent connective tissue entity, which was concomitant with a stratified epithelial equivalent. Immunohistochemistry proved tissue authenticity over time by abundance of the biomarker vimentin in the connective tissue entity, and chronological increase of keratins KRT1/10 and involucrin expression in epithelial equivalents. Suitability of the novel NBM as wound dressing was evidenced by an almost completion of epithelial wound closure in a pilot mini-pig study, after a surgical intervention-caused gingival dehiscence. In summary, preclinical assessment by tissue-authentic cell systems and the animal pilot study revealed the NBM as an encouraging therapeutic medical device for prospective clinical applications.

Biologic width adjacent to loaded implants with machined and rough collars in the dog.

Dental implant surface technology has evolved from a relatively smooth machined implant surface for osseointegration to more roughened osteoconductive surfaces. Recent studies suggest that peri-implant soft tissue inflammation with progressive bone loss (ie, peri-implantitis) is becoming a prevalent condition. One possibility that could explain such a finding is that more bacterial plaque forms on the roughened implant and abutment surfaces, which may result in the peri-implant inflammation in the soft tissues. This study compared 36 tissue-level implants with a machined transmucosal collar to 36 implants with a relatively roughened (SLActive) transmucosal surface in the dog. The implants were evaluated histologically and histomorphometrically after 3 and 12 months of loading. The results demonstrated that the connective tissue contact was similar between the two implant types but that the junctional epithelium and biologic width dimensions were greater around the implants with the machined collars. Interestingly, the amount of inflammation was similar between the two implant types. Slightly more bone formation and more mature collagen formation occurred around the implants with the roughened collars compared to the implants with machined collars. These results suggested that even if more plaque biofilm forms on the implants with the roughened SLActive surface compared to the machined surface, there is no biologic consequence related to the amount of inflammation or bone loss. In fact, the roughened surface promoted bone formation (was more osteoconductive) and more mature soft collagenous connective tissue.

Use of a perforated scaffold-retaining abutment to achieve vertical bone regeneration around dental implants in the minipig.

OBJECTIVE: To evaluate the effect of perforated scaffold retainers used in conjunction with dental implants and osteoinductive scaffolds to regenerate vertical supracrestal alveolar bone in an intraoral minipig model. MATERIALS AND METHODS: Three months after extraction of mandibular premolars and first molars from six adult minipigs, two titanium (Ti) custom implants were placed bilaterally in the edentulous mandibles for a total of four implants per animal. The upper 2.5 mm of the implant was left above bone level and covered with: (1) wide-neck healing caps; (2) perforated, overhanging custom scaffold retainers (umbrellas); or (3) scaffold retainers and demineralized minipig bone allograft (DBM) and nonglycosylated bone morphogenetic protein 2 (ng/rhBMP-2)-treated implants. All constructs were submerged beneath soft tissue flaps for 8 weeks. Two dental implant surfaces were compared: SLA and SLActive. Samples were retrieved after 8 weeks and analyzed by radiography, micro-computed tomography and histomorphometry. RESULTS: All implants were stable at the end of the experiment. Histomorphometry revealed that the use of the scaffold-retaining umbrellas led to increased, but not statistically significant, vertical bone regeneration as compared to the use of wide-neck healing caps (1.0 ± 0.4 mm vs 0.6 ± 0.3 mm). The combination of DBM and ng/rhBMP-2 released from the surface of the SLA implant resulted in the greatest amount of vertical bone regeneration (2.1 ± 0.2 mm). The bone-to-implant contact was similar for all groups. Mucosal dehiscence areas with healing cap or custom scaffold retainer exposures were reduced in the presence of ng/rhBMP-2. CONCLUSIONS: The combined use of custom perforated Ti scaffold retainers, DBM, and ng/rhBMP-2 regenerated a substantial amount of vertical supracrestal alveolar bone around Ti implants in an intraoral minipig model.

Evaluation of a new titanium-zirconium dental implant: a biomechanical and histological comparative study in the mini pig.

BACKGROUND: Titanium zirconium alloy with 13-17% zirconium (TiZr1317) shows significantly better mechanical attributes than pure Ti with respect to elongation and fatigue strength. This material may be suitable for thin implants and implant components exposed to high mechanical constraints. PURPOSE: The aim of this study was to test the hypothesis that TiZr1317 and Ti implants show comparable osseointegration and stability. MATERIALS AND METHODS: The mandibular premolars (P1, P2, P3) and the first molar (M1) in 12 adult miniature pigs were extracted 3 months prior to the study. Six specially designed implants made from Ti (commercially pure, Grade 4) or TiZr1317 (Roxolid®, Institut Straumann AG, Basel, Switzerland) with a hydrophilic sandblasted and acid-etched (SLActive, Institut Straumann AG, Basel, Switzerland) surface were placed in each mandible; three standard implants modified for evaluation of removal torque (RT) in one side and three bone-chamber implants for histologic observations in the contralateral side. RT tests were performed after 4 weeks when also the bone chamber implants and surrounding tissue were biopsied for histologic analyses in ground sections. RESULTS: The RT results indicated significantly higher stability (p=0.013) for TiZr1317 (230.9±22.4Ncm) than for Ti implants (204.7±24.0Ncm). The histology showed similar osteoconductive properties for both implant types. Histomorphometric measurements showed a statistically significant higher (p=0.023) bone area within the chamber for the TiZr1317 implants (45.5±13.2%) than did the Ti implants (40.2±15.2%). No difference was observed concerning the bone to implant contact between the groups with 72.3±20.5% for Ti and 70.2±17.3% for TiZr1317 implants. CONCLUSION: It is concluded that the TiZr1317 implant with a hydrophilic sandblasted and acid-etched surface showed similar or even stronger bone tissue responses than the Ti control implant.

Tissue integration of a new titanium-zirconium dental implant: a comparative histologic and radiographic study in the canine.

BACKGROUND: This study evaluates a newly developed titanium-zirconium implant (TiZr), comparing it to a commercially available pure titanium (Ti) implant subjected to the same surface treatment. METHODS: In nine dogs, 12 implants (six TiZr and six Ti) were randomly placed in the mandible with the implant shoulder at the bone crest and subjected to submerged healing. Standardized radiographs were taken after implantation, and at the sacrifice of 2 weeks (three dogs), 4 weeks (three dogs), and 8 weeks (three dogs). Histologic and histomorphometric measurements were performed on non-decalcified histologic sections. The main outcome measures included the first bone-implant contact (fBIC) and BIC over time. For statistical analysis, Wilcoxon signed-rank test and mixed model regressions were applied. RESULTS: From baseline to 8 weeks, a mean bone loss of 0.09 ± 0.33 mm for TiZr and a gain of 0.02 ± 0.33 mm for Ti were calculated radiographically. The number of implants with the fBIC coronal to the reference point (implant shoulder) gradually increased over time, reaching 39% of all TiZr implants and 50% of all Ti implants at 8 weeks. The mean fBIC values for Ti and TiZr were 0.29 ± 0.42 mm and 0.26 ± 0.32 mm (2 weeks), -0.01 ± 0.20 mm and 0.10 ± 0.28 mm (4 weeks), and -0.06 ± 0.22 mm and 0.08 ± 0.30 mm (8 weeks), respectively. The mean BIC values peaked at 86.9% ± 6.8% (8 weeks) for TiZr and at 83.4% ± 5.9% (4 weeks) for Ti. No statistically significant differences were observed at any time point. CONCLUSION: TiZr and Ti bone level implants with chemically-modified, sandblasted, and acid-etched surfaces performed similarly in regards to osseointegration in this unloaded canine study.

Clinical applications of glass-ceramics in dentistry.

Glass-ceramics featuring special properties can be used as a basis to develop biomaterials. It is generally differentiated between highly durable biomaterials for restorative dental applications and bioactive glass-ceramics for medical use, for example, bone replacements. In detail, this paper presents one biomaterial from each of these two groups of materials. In respect to the restorative dental biomaterials, the authors give an overview of the most important glass-ceramics for clinical applications. Leucite, leucite-apatite, lithium disilicate and apatite containing glass-ceramics represent biomaterials for these applications. In detail, the authors report on nucleation and crystallization mechanisms and properties of leucite-apatite glass-ceramics. The mechanism of apatite nucleation is characterized by a heterogeneous process. Primary crystal phases of alpha - and beta -NaCaPO4 were determined. Rhenanite glass-ceramics represent biomaterials with high surface reactivity in simulated body fluid, SBF, and exhibit reactive behaviour in tests with bone cells. Cell adhesion phenomena and cell growth were observed. Suitable colonization and proliferation and differentiation of cells as a preliminary stage in the development of a material for bone regeneration applications was established. The authors conclude that the processes of heterogeneous nucleation and crystallization are important for controlling the required reactions in both biomaterial groups.