Histological evaluation of osseointegration between conventional and novel bone-level tapered implants in healed bone-A preclinical study.

AIMS: To histologically compare osseointegration and crestal bone healing between newly introduced tapered, self-cutting bone-level test implants and tapered bone-level control implants in sites with fully healed sites. METHODS: Sixty-six implants (33 test, 33 control) were placed 1 mm subcrestally in a minipig model and underwent qualitative histologic and quantitative histometric analyses after 3, 6 and 12 weeks of submerged healing. The primary and secondary outcomes were the bone-to-implant contact (BIC) and first bone-to-implant contact (fBIC). Outcomes between the test and control implants were statistically compared. RESULTS: The BIC values of the test implants were comparable and non-inferior over the time points studied, except for the 12 weeks time point which showed statistically significantly higher BIC values of the test (88.07 ± 5.35%) compared to the control implants (80.88 ± 7.51%) (p = .010). Similarly comparable and non-inferior were the fBIC values, except for the 6-week outcome, which showed statistically higher values for the test (-546.5 ± 450.80 µm) compared to the control implants (-75.7 ± 100.59 µm). fBIC results for the test implants were qualitatively more stable and consistent between test time points. CONCLUSION: Novel self-cutting bone-level test implants demonstrated superior osseointegration and similar bone levels compared to conventional bone-level implants after a healing period of 12 weeks in healed ridges.

Determining primary stability for adhesively stabilized dental implants.

OBJECTIVES: To examine factors influencing the primary stability of dental implants when stabilized in over-sized osteotomies using a calcium phosphate-based adhesive cement was the objective. METHODS: Using implant removal torque measurements as a surrogate for primary stability, we examined the influence of implant design features (diameter, surface area, and thread design), along with cement gap size and curing time, on the resulting primary implant stability. RESULTS: Removal torque values scaled with implant surface area and increasing implant diameters. Cement gap size did not alter the median removal torque values; however, larger gaps were associated with an increased spread of the measured values. Among the removal torque values measured, all were found to be above 32 Ncm which is an insertion torque threshold value commonly recommended for immediate loading protocols. CONCLUSION: The adhesive cement show potential for offering primary implant stability for different dental implant designs. In this study, the primary parameters influencing the measured removal torque values were the implant surface area and diameter. As the liquid cement prevents the use of insertion torque, considering the relationship between insertion and removal torque, removal torque can be considered a reliable surrogate for primary implant stability for bench and pre-clinical settings. CLINICAL RELEVANCE: At present, the primary stability of dental implants is linked to the quality of the host bone, the drill protocol, and the specific implant design. The adhesive cement might find applications in future clinical settings for enhancing primary stability of implants under circumstances where this cannot be achieved conventionally.

Immediate loading of a fully tapered implant with deep apical threads placed in healed alveolar ridges vs. immediate extraction sockets.

OBJECTIVES: Immediate implant placement and loading is a practice that continues to gain traction in implant dentistry because it reduces treatment time and improves satisfaction. Novel implant designs that facilitate increased primary stability, while not compromising osseointegration and long-term survival are important to offer immediate solutions for missing teeth. Here, we hypothesize that fully tapered implants can obtain successful osseointegration with high survival rates after immediate loading in fresh extraction sockets and healed sites. MATERIALS AND METHODS: A total of 13 swine with 73 implants were evaluated. Fully tapered or apically tapered implants were placed in extraction sockets and healed sites. Insertion torque and resonance frequency analysis were determined at placement and euthanasia. Animals were evaluated at: placement, and 1-week and 12-weeks after placement. Bone to Implant Contact (BIC), Bone Area/Total Area (BA/TA), and first BIC (fBIC) analyses were conducted. RESULTS: The fully tapered implant achieved similar primary stability with lower insertion torque at placement. Apically and fully tapered implants had comparable BIC (50.1% vs 59.4%) and ISQ (82.5 vs 80.3) values by 12 weeks in healed sites. In extraction sockets, BIC and ISQ for the apically tapered implant was 35.8% and 73.2 and 37.8% and 79.2 for the fully tapered implants, respectively. CONCLUSIONS: In this short-term study, immediately loaded fully tapered implants obtained high survival with similar osseointegration ability as apically tapered implants when placed in healed sites and fresh extraction sockets. Fully tapered implants show promise for use in immediate loading and immediate placement.

Microtomographic reconstruction of mandibular defects treated with xenografts and collagen-based membranes: A pre-clinical minipig model / Reconstrucción microtomográfica de defectos mandibulares tratados con Xenoinjertos y membranas a base de colágeno: un modelo preclínico de minicerdo

Background: The goal of this study was to evaluate hard tissue response following guided bone regeneration us-ing commercially available bovine bone grafts and collagen membranes; bilayer collagen membrane and porcinepericardium-based membrane, by means of a non-destructive three-dimensional (3D) computerized volumetricanalysis following microtomography reconstruction.Material and Methods: Bone regenerative properties of various bovine bone graft materials were evaluated in theGöttingen minipig model. Two standardized intraosseous defects (15mm x 8mm x 8mm) were created bilaterallyof the mandible of eighteen animals (n=72 defects). Groups were nested within the same subject and randomlydistributed among the sites: (i) negative control (no graft and membrane), (ii) bovine bone graft/bilayer collagenmembrane (BOB) (iii) Bio-Oss® bone graft/porcine pericardium-based membrane (BOJ) and (iv) cerabone® bonegraft/porcine pericardium-based membrane (CJ). Samples were harvested at 4, 8, and 12-week time points (n=6animal/time point). Segments were scanned using computerized microtomography (μCT) and three dimensionallyreconstructed utilizing volumetric reconstruction software. Statistical analyses were performed using IBM SPSSwith a significance level of 5% Results: From a temporal perspective, tridimensional evaluation revealed gradual bone ingrowth with the presenceof particulate bone grafts bridging the defect walls, and mandibular architecture preservation over time. Volumetricanalysis demonstrated no significant difference between all groups at 4 weeks (p>0.127). At 8 and 12 weeks therewas a higher percentage of new bone formation for control and CJ groups when compared to BOB and BOJ groups(p<0.039). The natural bovine bone graft group showed more potential for graft resorption over time relative to bo-vine bone graft, significantly different between 4 and 8 weeks (p<0.003)...(AU)

Comparative barrier membrane degradation over time: Pericardium versus dermal membranes.

OBJECTIVE: The effectiveness of GBR procedures for the reconstruction of periodontal defects has been well documented. The objective of this investigation was to evaluate the degradation kinetics and biocompatibility of two resorbable collagen membranes in conjunction with a bovine xenograft material. MATERIALS AND METHODS: Lower premolars and first molars were extracted from 18 male Yucatan minipigs. After 4 months of healing, standardized semi-saddle defects were created (12 mm × 8 mm × 8 mm [l˙̇ × W˙ × d]), with 10 mm between adjacent defects. The defects were filled with a bovine xenograft and covered with a either the bilayer collagen membrane (control) or the porcine pericardium-derived collagen membrane (test). Histological analysis was performed after 4, 8, and 12 weeks of healing and the amount of residual membrane evaluated. Non-inferiority was calculated using the Brunner-Langer mixed regression model. RESULTS: Histological analysis indicated the presence of residual membrane in both groups at all time points, with significant degradation noted in both groups at 12 weeks compared to 4 weeks (p = .017). No significant difference in ranked residual membrane scores between the control and test membranes was detected at any time point. CONCLUSIONS: The pericardium-derived membrane was shown to be statistically non-inferior to the control membrane with respect to resorption kinetics and barrier function when utilized for guided bone regeneration in semi-saddle defects in minipigs. Further evaluation is necessary in the clinical setting.

Peri-implant bone preservation of a novel, self-cutting, and fully tapered implant in the healed crestal ridge of minipigs: submerged vs. transgingival healing.

OBJECTIVES: The aim of this study was to assess the influence of transgingival compared with submerged healing on peri-implant bone maintenance around a novel, fully tapered implant in a healed crestal ridge in minipigs. MATERIALS AND METHODS: In each of 12 minipigs, two implants (Straumann® BLX, Roxolid® SLActive®, Ø 3.75 × 8 mm) were placed. Implants were either left for submerged or for transgingival healing for 12 weeks. Measurements performed were bone-to-implant contact (BIC), first bone-to-implant contact (fBIC), bone area to total area (BATA), perpendicular bone crest to implant shoulder (pCIS), bone height change from placement, and bone overgrowth (for submerged implants). RESULTS: No significant differences were found between transgingival and submerged healing in any of the measured parameters, except for BATA on the buccal aspect in which significantly more bone formation was found for the transgingival healing group. For both groups, there was a gain in crestal bone height during the 12-week healing period. CONCLUSIONS: Loaded compared with unloaded implants displayed comparable levels of osseointegration and equivalent marginal bone levels. This qualifies the implant placement protocol with respect to the osteotomy dimensions and subcrestal placement protocol for immediate loading. CLINICAL RELEVANCE: The here presented results related to osseointegration and crestal bone maintenance after submerged or transgingival healing have demonstrated a high level of consistency in the used in vivo translational model. The obtained results support the translation of the novel implant type in conjunction with the developed surgical workflow and placement protocol into further clinical investigation and use.

Greater Osseointegration Potential with Nanostructured Surfaces on TiZr: Accelerated vs. Real-Time Ageing.

Surface chemistry and nanotopography of dental implants can have a substantial impact on osseointegration. The aim of this investigation was to evaluate the effects of surface chemistry and nanotopography on the osseointegration of titanium-zirconium (TiZr; Roxolid®) discs, using a biomechanical pull-out model in rabbits. Two discs each were placed in both the right and left tibiae of 16 rabbits. Five groups of sandblasted acid etched (SLA) discs were tested: (1) hydrophobic without nanostructures (dry/micro) (n = 13); (2) hydrophobic with nanostructures, accelerated aged (dry/nano/AA) (n = 12); (3) hydrophilic without nanostructures (wet/micro) (n = 13); (4) hydrophilic with nanostructures, accelerated aged (wet/nano/AA; SLActive®) (n = 13); (5) hydrophilic with nanostructures, real-time aged (wet/nano/RTA). The animals were sacrificed after four weeks and the biomechanical pull-out force required to remove the discs was evaluated. Adjusted mean pull-out force was greatest for group wet/nano/RTA (64.5 ± 17.7 N) and lowest for group dry/micro (33.8 ± 10.7 N). Multivariate mixed model analysis showed that the pull-out force was significantly greater for all other disc types compared to the dry/micro group. Surface chemistry and topography both had a significant effect on pull-out force (p < 0.0001 for both), but the effect of the interaction between chemistry and topography was not significant (p = 0.1056). The introduction of nanostructures on the TiZr surface significantly increases osseointegration. The introduction of hydrophilicity to the TiZr implant surface significantly increases the capacity for osseointegration, irrespective of the presence or absence of nanotopography.

A novel fully tapered, self-cutting tissue-level implant: non-inferiority study in minipigs.

OBJECTIVES: To assess the osseointegration and crestal bone level maintenance of a novel fully tapered self-cutting tissue-level implant for immediate placement (test) compared to a clinically established tissue-level implant (control) in moderate bone quality. MATERIALS AND METHODS: Test and control implants were compared in 3 groups, i.e., small-, medium-, and large-diameter implants in an edentulous mandibular minipig model with moderate bone quality after 12 weeks of healing. Histometrically derived bone-to-implant contact (BIC) and first bone-to-implant contact (fBIC) were subjected to statistical non-inferiority testing. Maximum insertion torque values in artificial bone were assessed for comparison. RESULTS: BIC values for the tests and control implants for all 3 diameters were comparable and non-inferior: small diameter (61.30 ± 10.63% vs. 54.46 ± 18.31%) (p=0.99), medium diameter (60.91 ± 14.42 vs. 54.68 ± 9.16) (p=0.55), and large diameter (45.60 ± 14.67 vs. 52.52 ± 14.76) (p=0.31). fBIC values for test implants were higher and non-inferior compared to control implants in all three groups. Test implants further showed distinctly higher maximum insertion torque values compared to control implants. CONCLUSION: The investigated novel tissue-level implant is able to achieve high levels of primary and secondary implant stability under simultaneous preservation of crestal bone levels. This qualifies the studied implant as an attractive candidate for immediate placement in bone of limited quality. CLINICAL RELEVANCE: This pilot pre-clinical study investigated a novel tissue-level implant for immediate placement. With the aim of translating the studied prototype into clinical application pre-clinical models, procedures and controls have been chosen with the aim of reflecting its future clinical indication and use.

Impact of Implant Surface Material and Microscale Roughness on the Initial Attachment and Proliferation of Primary Human Gingival Fibroblasts.

Due to the rising demand for zirconia (Zr) based implant systems, it is important to understand the impact of Zr and titanium (Ti) implants and particularly their topography on soft tissue healing. As human gingival fibroblasts (hGFs) are the predominant cells in peri-implant soft tissue, we focused on examining the effect of implant material and surface roughness on hGFs' initial attachment, growth and the expression of proteins involved in the focal adhesion. hGFs isolated from eight healthy donors were cultured on the following surfaces: smooth titanium machined surface (TiM), smooth zirconia machined surface (ZrM), moderately rough titanium surface (SLA), or moderately rough zirconia surface (ZLA) for up to 14 days. The initial attachment of hGFs was evaluated by scanning electron microscopy. Cell proliferation/viability was assessed by cell counting kit 8. Focal adhesion and cytoskeleton were visualized by a focal adhesion staining kit. The gene expression of focal adhesion kinase (FAK), α-smooth muscle actin (α-SMA), and integrin subunits ITG-ß1, ITG-ß4, ITG-α4, ITG-α5, ITG-α6, was evaluated by qPCR. Cell proliferation/viability was slightly decreased by moderately rough surfaces, whereas no effect of surface material was observed. Cell morphology was strikingly different between differently treated surfaces: on machined surfaces, cells had elongated morphology and were attached along the grooves, whereas on moderately rough surfaces, cells were randomly attached. Surface roughness had a more pronounced effect on the gene expression compared to the surface material. The expression of FAK, α-SMA, ITG-ß4, ITG-α5, and ITG-α6 was enhanced by moderately rough surfaces compared to smooth surfaces. Within the limitations of this in vitro study, it can be concluded that the behavior of primary hGFs is primarily affected by surface structure, whereas no apparent advantage of Zr over Ti could be observed.

Thermal exposure of implant osteotomies and its impact on osseointegration-A preclinical in vivo study.

OBJECTIVES: Thermal and mechanical stresses during osteotomy preparation can impair implant osseointegration. This study investigated implant osseointegration following the measurement of temperature exposure during osteotomy drilling, varying drill design, sequence, and drill wear. MATERIALS AND METHODS: 36 tapered implants were placed in a mandibular minipig model after guided drilling of implant osteotomies using 4 different groups: (1) control drills with a conservative, sequential drilling sequence, (2) control drills using a shortened drill sequence (PF), (3) novel test drill displaying an optimized drill design and surface treatment, PF, and (4) aged test drill, PF. Intraosseous temperatures during drilling were measured using a temperature probe. BIC, fBIC, and tissue reactions were histomorphometrically derived after 2 and 8 weeks of healing. RESULTS: Compared to control drills (1) or (2), test drills (3) resulted in significantly lower maximum temperatures ((35.4 (CI 30.2-40.5)°C vs. (46.5 (CI 41.0-52.0)°C, p = .0021)) and shorter drill times ((4.5 (CI 1.6-7.3)sec vs. 10.3 (7.3-13.4)sec). Lower osteotomy temperature values and shorter drill times corroborated with significantly higher BIC after 2 and 8 weeks healing for the test (3) compared to control groups (2) (2 weeks: (44.9 (CI 34.1-55. 7)% vs. (31.3 (CI 20.5-42.2)%, p = <.0001 and 8 weeks: (73.7 ( CI 64.2-83.2)% vs. (66.2 (CI 57.0-75.4)%, p = <.0455). CONCLUSION: The improved osseointegration of implants placed after osteotomy preparation with novel test drills using a shortened drill sequence compared to standard drills and conventional drill protocols might be attributed to more favorable thermal profiles and less mechanical stress exerted on the bone surrounding the implant osteotomy.

Evaluation of air polishing with a sterile powder and mechanical debridement during regenerative surgical periimplantitis treatment: a study in dogs.

OBJECTIVES: To evaluate the effectiveness of mechanical debridement and/or air polishing on the healing of ligature-induced buccal periimplantitis dehiscence defects in dogs. MATERIAL AND METHODS: Forty-eight implants were placed in the mandibles of twelve beagle dogs, and periimplantitis was induced for 2 months using ligatures. The resulting buccal dehiscence-type defects were surgically cleaned and augmented (xenogenic filler and resorbable membrane) according to one of the following treatments: (1) Cleaning with carbon curette (debridement - D) and guided bone regeneration (GBR/G): DG, (2) air polishing cleaning (A) and GBR: AG, (3) a combination of D/A/G: DAG, and (4) D/A without GBR: DA. After 2 months, histomorphometric and inflammatory evaluations were conducted. RESULTS: The median bone gain after therapy ranged between 1.2 mm (DG) and 2.7 mm (AG). Relative bone gain was between 39% (DG) and 59% (AG). The lowest inflammation scores were obtained in DA without GBR (5.84), whereas significantly higher values between 8.2 and 9.4 were found in the groups with augmentation. At lingual sites without defects, scores ranged from 4.1 to 5.9. According to ISO, differences above 2.9 were considered representative for irritative properties. CONCLUSIONS: All treatments resulted in partial regeneration of the defects. No treatment group showed a significantly (p < 0.05) better outcome. However, pretreatment with air polishing showed a tendency for less inflammation. Noteworthy, inflammation assessment showed an overall irritative potential after GBR in the evaluated early healing phase. CLINICAL RELEVANCE: Periimplantitis treatment still represents a big issue in daily practice and requires additional preclinical research in order to improve treatment concepts.

Osseointegration of a novel injection molded 2-piece ceramic dental implant: a study in minipigs.

OBJECTIVES: This study compared the osseointegrative potential of a novel injection molded zirconia dental implant (Neodent Zi ceramic implant, test) and a commercially available titanium implant (Neodent Alvim implant, control) in terms of histomorphometrically derived bone-to-implant contact (BIC), first bone-to-implant contact (fBIC), and the ratio of bone area to total area (BATA) around the implant. MATERIALS AND METHODS: A total of 36 implants, 18 per individual test device, were implanted in a split-mouth arrangement in either side of the edentulous and fully healed mandible of 6 minipigs. Histomorphometric analysis of BIC, fBIC, and BATA were performed 8 weeks post implantation and subjected to statistical non-inferiority testing. Surface characteristics of both implant types were compared in terms of contact angle, surface topography, and elemental composition. RESULTS: BIC, fBIC, and coronal BATA values of test and control implants were statistically comparable and non-inferior. BIC values of 77.8 ± 6.9% vs. 80.7 ± 6.9% (p = 0.095) were measured for the test and control groups. fBIC lingual values were - 238 ± 328 µm compared with - 414 ± 511 µm (p = 0.121) while buccal values were - 429 ± 648 µm and - 588 ± 550 µm (p = 0.230) for the test and control devices, respectively. BATA in the apical segment was significantly higher in the test group compared with the control group (67.2 ± 11.8% vs. 59.1 ± 11.4%) (p = 0.0103). Surface topographies of both implant types were comparable. Surface chemical analysis indicated the presence of carbonaceous adsorbates which correlated with a comparable and predominantly hydrophobic character of the implants. CONCLUSION: The results demonstrate that the investigated zirconia implants, when compared with a commercially available titanium implant, show equivalent and non-inferior bone integration, bone formation, and alveolar bone level maintenance. This qualifies the investigated zirconia implant as a potential candidate for clinical development. CLINICAL RELEVANCE: This study investigated the osseointegration of a novel zirconia 2-piece dental implant prototype intended for clinical development. With the aim of translating this prototype into clinical development preclinical models, procedures and materials within this study have been selected as close to clinical practice and human physiological conditions as possible.

A Novel Xenograft Bone Substitute Supports Stable Bone Formation in Circumferential Defects Around Dental Implants in Minipigs.

PURPOSE: The aim of this study was to evaluate and compare bone growth and implant integration in circumferential defects with two commercially available bone substitutes (demineralized bovine bone mineral [DBBM]). MATERIALS AND METHODS: Circumferential defects were created in the mandibles of minipigs (n = 10), and Bone Level Tapered implants (Straumann Roxolid with SLActive surface) were placed. The defects (4-mm-deep circumferential defect, 2 mm around each implant) were augmented with either sintered bovine bone mineral (test, cerabone) or natural bovine bone mineral (control, Bio-Oss). Bone formation and tissue composition in augmented sites were histomorphometrically assessed after 8 and 12 weeks of healing time (n = 5 each), respectively, in terms of the percentage of area of newly formed bone to total area, bone-to-implant contact (BIC), and crestal bone height relative to the implant shoulder (first bone-to-implant contact [fBIC]). RESULTS: Bone formation in all defect sites was adequate and equivalent for both groups at individual healing time points. The amount of residual graft material was comparable in both groups after 8 and 12 weeks, with no significant resorption in either group. The mean newly formed bone area in the test group amounted to 46.7% ± 5.1% and 48.7% ± 4.0% after 8 and 12 weeks vs 47.0% ± 4.8% and 47.8% ± 7.3% in the control group, respectively. BIC and fBIC as individually assessed for the lingual and buccal aspects were comparable at both healing time points without any statistically significant differences between the groups. A slightly greater variability of fBIC was observed within the test group. CONCLUSION: The results of this study indicate that test and control materials both represent viable bovine bone graft material that equivalently support the formation of new and stable bone volume specifically when used for simultaneous augmentation around implants.

Surface modification of ultrafine-grained titanium: Influence on mechanical properties, cytocompatibility, and osseointegration potential.

OBJECTIVE: The main objective of this study was to demonstrate that dental implants made from ultrafine-grain titanium (UFG-Ti) can be created that replicate state of the art surfaces of standard coarse-grain titanium (Ti), showing excellent cytocompatibility and osseointegration potential while also providing improved mechanical properties. MATERIAL AND METHODS: UFG-Ti was prepared by continuous equal channel angular processing (ECAP), and surfaces were treated by sandblasting and acid etching. Mechanical properties (tensile and fatigue strength), wettability, and roughness parameters were evaluated. Human trabecular bone-derived osteoblast precursor cells (HBCs) were cultured on all samples to examine cytocompatibility and mineralization after 4 and 28 days, respectively. Biomechanical pull-out measurements were performed in a rabbit in vivo model 4 weeks after implantation. RESULTS: Both yield and tensile strength as well as fatigue endurance were higher for UFG-Ti compared to Ti by 40%, 45%, and 34%, respectively. Fatigue endurance was slightly reduced following surface treatment. Existing surface treatment protocols could be applied to UFG-Ti and resulted in similar roughness and wettability as for standard Ti. Cell attachment and spreading were comparable on all samples, but mineralization was higher for the surfaces with hydrophilic treatment with no significant difference between UFG-Ti and Ti. Pull-out tests revealed that osseointegration of surface-treated UFG-Ti was found to be similar to that of surface-treated Ti. CONCLUSION: It could be demonstrated that existing surface treatments for Ti can be translated to UFG-Ti and, furthermore, that dental implants made from surface-treated UFG-Ti exhibit superior mechanical properties while maintaining cytocompatibility and osseointegration potential.

Ascorbic Acid Attenuates Senescence of Human Osteoarthritic Osteoblasts.

The accumulation of senescent cells is implicated in the pathology of several age-related diseases. While the clearance of senescent cells has been suggested as a therapeutic target for patients with osteoarthritis (OA), cellular senescence of bone-resident osteoblasts (OB) remains poorly explored. Since oxidative stress is a well-known inducer of cellular senescence, we here investigated the effect of antioxidant supplementation on the isolation efficiency, expansion, differentiation potential, and transcriptomic profile of OB from osteoarthritic subchondral bone. Bone chips were harvested from sclerotic and non-sclerotic regions of the subchondral bone of human OA joints. The application of 0.1 mM ascorbic acid-2-phosphate (AA) significantly increased the number of outgrowing cells and their proliferation capacity. This enhanced proliferative capacity showed a negative correlation with the amount of senescent cells and was accompanied by decreased expression of reactive oxygen species (ROS) in cultured OB. Expanded cells continued to express differentiated OB markers independently of AA supplementation and demonstrated no changes in their capacity to osteogenically differentiate. Transcriptomic analyses revealed that apoptotic, cell cycle-proliferation, and catabolic pathways were the main pathways affected in the presence of AA during OB expansion. Supplementation with AA can thus help to expand subchondral bone OB in vitro while maintaining their special cellular characteristics. The clearance of such senescent OB could be envisioned as a potential therapeutic target for the treatment of OA.

In vitro evaluation of an injectable biphasic calcium phosphate (BCP) carrier system combined with recombinant human bone morphogenetic protein (rhBMP)-9.

Bone morphogenetic protein 9 (BMP9) has previously been characterized as the strongest osteoinductive growth factor among the BMP family. The aim of the present study was to evaluate the possibility of combining rhBMP9 with an injectable biphasic calcium phosphate (I-BCP, maxresorb inject®), since I-BCP is an easy to handle biomaterial with ideal properties for bone augmentation procedures. The adsorption potential of rhBMP9 as well as the cell behavior of bone stromal ST2 cells were investigated on cell viability, adhesion, proliferation and osteogenic differentiation for I-BCP combined with/without rhBMP9 in vitro. I-BCP demonstrated excellent adsorption/retention potential of rhBMP9 with a slow and steady release over a 10 day period by ELISA. Cell attachment at 8 hours and cell proliferation at 1, 3 and 5 days was decreased on I-BCP with/without rhBMP9 when compared to control tissue-culture plastic. While I-BCP had little influence on osteoblast differentiation, its combination with rhBMP9 significantly increased ALP activity at 7 days and mRNA levels of osteoblast differentiation markers including ALP and osteocalcin at 14 days. I-BCP served as an excellent carrier for rhBMP9 clearly demonstrating its osteoinductive potential. We therefore confirm the great potential of rhBMP9 to serve as a future regenerative growth factor for bone applications.

Absorbable collagen sponges loaded with recombinant bone morphogenetic protein 9 induces greater osteoblast differentiation when compared to bone morphogenetic protein 2.

The use of growth factors for the regeneration of soft and hard tissues has been utilized extensively in dental medicine over the past decade. Recently our group found that recombinant human bone morphogenetic protein 9 (rhBMP9) was more osteopromotive than recombinant human bone morphogenetic protein 2 (rhBMP2) when combined with a deprotenized bovine bone mineral bone grafting material. The aim of the present in vitro study was to evaluate the regenerative potential of an absorbable collagen sponge(ACS) specifically designed for extraction socket healing loaded with rhBMP9 when compared to rhBMP2. The adsorption and release kinetics of rhBMP2 and rhBMP9 were first investigated by enzyme-linked immunosorbent assay quantification. Then, the cellular effects of stromal cell line (ST2) preosteoblasts were investigated utilizing four groups including rhBMP2 and rhBMP9 at both low(10 ng/ml) and high(100 ng/ml) concentrations loaded onto ACS. Cellular attachment(8 hours) and proliferation(1, 3, and 5 days) as well as osteoblast differentiation were investigated by real-time polymerase chain reaction (PCR) at 3 and 14 days, alkaline phosphatase (ALP) activity at 7 days, and alizarin red staining at 14 days. ACS fully adsorbed both rhBMP2 and rhBMP9 that were slowly released up to 10 days. Although neither rhBMP2 nor rhBMP9 had any effects on cell attachment or proliferation, pronounced effects were observed on osteoblast differentiation. ALP activity was increased seven-fold with rhBMP2-high, whereas a marked 10-fold and 20-fold increase was observed with rhBMP9-low and high loaded to ACS, respectively. Furthermore, mRNA levels of collagen1, ALP, bone sialoprotein, and osteocalcin were all significantly higher for rhBMP9 when compared to control or rhBMP2 groups. Alizarin red staining further confirmed that rhBMP9-low and high demonstrated marked increases in mineralization potential when compared to rhBMP2-high. The results demonstrate the marked effect of rhBMP9 on osteoblast differentiation when combined with ACS in comparison to rhBMP2 at doses as much as 10 times lower. Further in vivo studies are necessary to investigate whether the regenerative potential is equally as potent.

Pooled thrombin-activated platelet-rich plasma: a substitute for fetal bovine serum in the engineering of osteogenic/vasculogenic grafts.

The use of fetal bovine serum (FBS) as a culture medium supplement in cell therapy and clinical tissue engineering is challenged by immunological concerns and the risk of disease transmission. Here we tested whether human, thrombin-activated, pooled, platelet-rich plasma (tPRP) can be substituted for FBS in the engineering of osteogenic and vasculogenic grafts, using cells from the stromal vascular fraction (SVF) of human adipose tissue. SVF cells were cultured under perfusion flow into porous hydroxyapatite scaffolds for 5 days, with the medium supplemented with either 10% tPRP or 10% FBS and implanted in an ectopic mouse model. Following in vitro culture, as compared to FBS, the use of tPRP did not modify the fraction of clonogenic cells or the different cell phenotypes, but increased by 1.9-fold the total number of cells. After 8 weeks in vivo, bone tissue was formed more reproducibly and in higher amounts (3.7-fold increase) in constructs cultured with tPRP. Staining for human-specific ALU sequences and for the human isoforms of CD31/CD34 revealed the human origin of the bone, the formation of blood vessels by human vascular progenitors and a higher density of human cells in implants cultured with tPRP. In summary, tPRP supports higher efficiency of bone formation by SVF cells than FBS, likely by enhancing cell expansion in vitro while maintaining vasculogenic properties. The use of tPRP may facilitate the clinical translation of osteogenic grafts with intrinsic capacity for vascularization, based on the use of adipose-derived cells. Copyright © 2015 John Wiley & Sons, Ltd.

Dimensional Changes Following Immediate and Delayed Implant Placement: A Histomorphometric Study in the Canine.

PURPOSE: The placement of immediate implants has become a routine treatment modality for the replacement of missing teeth. The aim of the present study was to evaluate dimensional changes of hard tissues following tooth extraction, implant bed preparation, and the subsequent integration of titanium implants placed immediately or following an 8-week healing period. MATERIALS AND METHODS: All mandibular premolars were extracted and used as experimental sites in five beagle dogs. The left quadrant was used to compare the healing of alveolar bone following regular tooth extraction or surgical implant bed preparation. The right quadrant was used to compare immediate with delayed implant placement (3.3 mm, Bone Level SLActive, Roxolid, Straumann) after an 8-week healing period. All samples were assessed histologically for new bone area/total area (BATA), bone-to-implant contact (BIC), and bone height changes from the buccal and lingual midlines. RESULTS: Implant bed preparation led to significantly lower BATA values and bone height loss compared with regular extraction sockets (-2.46 mm vs -1.44 mm, respectively; P = .0007). Placement of immediate implants demonstrated buccal and lingual bone height loss of 1.51 mm and 0.91 mm, respectively, whereas only 0.12 mm and 0.12 mm of bone height loss was observed during delayed implant placement. A significantly higher BIC was observed for implants placed immediately compared with delayed placement (52% vs 35%, P < .05). CONCLUSION: Implant bed preparation further increased dimensional changes when compared with routine extraction sockets. Although immediate implant placement produced better BIC than did delayed placement, a significantly higher bone height loss at both the buccal and lingual midlines was reported. Further investigation is underway to minimize dimensional changes following immediate implant placement.

Osteoinductive potential of 4 commonly employed bone grafts.

OBJECTIVES: Guided bone regeneration (GBR) aims to predictably restore missing bone that has been lost due to trauma, periodontal disease or a variety of systemic conditions. Critical to this procedure is the ability of a bone grafting material to predictably serve as a 3-dimensional scaffold capable of inducing cell and bone tissue in-growth at the material surface. Although all bone grafts are osteoconductive to bone-forming osteoblasts, only a small number of commercially available bone grafts with FDA approval are osteoinductive including demineralized freeze-dried bone allographs (DFDBA) and scaffolds containing bone morphogenetic proteins (BMPs). Recently, a class of synthetic bone grafts fabricated from biphasic calcium phosphate (BCP) sintered at a low temperature have been shown to form ectopic bone formation in non-skeletal sites without the use of growth factors. Therefore, the present study aimed to compare the osteoinductive potential of this group of synthetic BCP alloplasts with autografts, allografts and xenografts. MATERIALS AND METHODS: In the present study, 4 types of bone grafting materials including autogenous bone harvested with a bone mill, DFDBA (LifeNet, USA), a xenograft derived from bovine bone mineral (NBM, BioOss, Geistlich, Switzerland) and a novel synthetic biphasic calcium phosphate (BCP, Straumman, Switzerland) were implanted into intramuscular pouches of 24 rats and analysed histologically for their ability to form ectopic bone formation around grafting particles. A semi-quantitative osteoinductive score was used to quantify the osteoinductive ability of each bone graft. RESULTS: The results from the present study reveal that (1) autogenous bone resorbed rapidly in vivo, (2) the xenograft showed no potential to form ectopic bone formation and (3) both DFDBA and BCP were able to stimulate ectopic bone formation. CONCLUSION: These studies demonstrate that these newly developed synthetic bone grafts have potential for inducing ectopic bone formation similar to DFDBA. Future clinical testing is necessary to reveal their bone-inducing properties in clinical scenarios including GBR procedures and in combination with implant dentistry. CLINICAL RELEVANCE: Novel BCP scaffolds are able to induce ectopic bone formation without the use of osteoinductive growth factors such as BMP2 and thus demonstrate a large clinical possibility to further enhance bone formation for a variety of clinical procedures.