Karl Andreas Schlegel

TiO2 Nanotube Surfaces: 15 nm—An Optimal Length Scale of Surface Topography for Cell Adhesion and Differentiation†

Studies of biomimetic surfaces in medicine and biomaterial fields have explored extensively how the micrometer-scale topography of a surface controls cell behavior, but only recently has the nanoscale environment received attention as a critical factor for cell behavior. Several investigations of cell interactions have been performed using surface protrusion topographies at the nanoscale; such topographies are typically based on polymer demixing, ordered gold cluster arrays, or islands of adhesive ligands at distinct length scales. Recent work has indicated that the fabrication of ordered TiO2 nanotube layers with controlled diameters can be achieved by anodization of titanium in adequate electrolytes. Such surfaces can almost ideally be used as nanoscale spacing models for size-dependent cellular response. This is particularly important as these studies are carried out on titanium surfaces—a material used for clinical titanium implantations for the purpose of bone, joint, or tooth replacements. Therefore, principles elucidated from this work can guide implant surface modifications toward an optimized surface geometry and profile to best fit and cell interactions for adequate bone growth.

In vivo evaluation of anodic TiO2 nanotubes: an experimental study in the pig.

Because of their ability to mimic the dimensions of constituent components of natural bone and the possibility to serve as a gene and drug-delivery carrier, nanotubes seem to be a promising coating for medical implants. Aim of this study was to investigate the effects of a TiO(2) nanotube structured surface on periimplant bone formation in vivo when compared with an untreated standard titanium surface. Twenty-five titanium implants covered with an ordered TiO(2) nanotube layer with an individual tube diameter of 30 nm and 25 commercially pure titanium (cp-Ti) implants were placed in the frontal skull of 25 domestic pigs. To evaluate the effects of the nanotube structured implants on the periimplant bone formation, bone-implant contact (BIC), and immunohistochemistry analysis were performed at day 3, 7, 14, 30, and 90. Evaluating immunohistochemistry, a significantly higher collagen type- I expression occurred at day 7 (p = 0.003), day 14 (p = 0.016), and day 30 (p = 0.044), for the nanostructured implants in comparison with the control group. It could be found that a nanotube structured implant surface with a diameter of 30 nm does influence bone formation and bone development by enhancing osteoblast function. SEM evaluation of the specimen surfaces revealed that the nanotube coatings do resist shearing forces that evoked by implant insertion. Because of their simple, low cost, flexible manufacturing and the possibility for the usage as drug or growth factor delivery system, nanotubes seem to be a promising method for future medical implant coatings.

In vivo performance of selective electron beam-melted Ti-6Al-4V structures.

Highly porous titanium structures are widely used for maxillofacial and orthopedic surgery because of their excellent mechanical properties similar to those of human bone and their facilitation of bone ingrowth. In contrast to common methods, the generation of porous titaniumproducts by selective electron beam melting (SEBM), an additive manufacturing technology, overcomes difficulties concerning the extreme chemical affinity of liquid titanium to atmospheric gases which consequently leads to strongly reduced ductility of the metal. The purpose of this study was to assess the suitability of a smooth compact and a porous Ti-6Al-4V structure directly produced by the SEBM process as scaffolds for bone formation. SEBM-processed titanium implants were placed into defects in the frontal skull of 15 domestic pigs. To evaluate the direct contact between bone and implant surfaces and to assess the ingrowth of osseous tissue into the porous structure, microradiographs and histomorphometric analyses were performed 14, 30, and 60 days after surgery. Bone ingrowth increased significantly during the period of this study. After 14 days the most outer regions of the implants were already filled with newly formed bone tissue (around 14%). After 30 days the bone volume inside the implants reached almost 30% and after 60 days abundant bone formation inside the implants attained 46%. During the study only scarce bone-implant contact was found around all implants, which did not exceed 9% around compact specimens and 6% around porous specimens after 60 days. This work demonstrates that highly porous titanium implants with excellent interconnectivity manufactured using the SEBM method are suitable scaffolds for bone ingrowth. This technique is a good candidate for orthopedic and maxillofacial applications.

Radiographic evaluation of marginal bone levels adjacent to parallel-screw cylinder machined-neck implants and rough-surfaced microthreaded implants using digitized panoramic radiographs.

OBJECTIVE The purpose of this split-mouth study was to compare macro- and microstructure implant surfaces at the marginal bone level during a stress-free healing period and under functional loading. MATERIAL AND METHODS From January to February 2006, 133 implants (70 rough-surfaced microthreaded implants and 63 machined-neck implants) were inserted in the mandible of 34 patients with Kennedy Class I residual dentitions and followed until February 2008. The marginal bone level was radiographically determined, using digitized panoramic radiographs, at four time points: at implant placement (baseline level), after the healing period, after 6 months of functional loading, and at the end of follow-up. RESULTS The median follow-up time was 1.9 (range: 1.9-2.1) years. The machined-neck group had a mean crestal bone loss of 0.5 mm (range: 0-2.3) after the healing period, 0.8 mm after 6 months (range: 0-2.4), and 1.1 mm (range: 0-3) at the end of follow-up. The rough-surfaced microthreaded implant group had a mean bone loss of 0.1 mm (range: -0.4-2) after the healing period, 0.4 mm (range: 0-2.1) after 6 months, and 0.5 mm (range: 0-2.1) at the end of follow-up. The two implant types showed significant differences in marginal bone levels (healing period: P=0.01; end of follow-up: P<0.01). CONCLUSIONS Radiographic evaluation of marginal bone levels adjacent to machined-neck or rough-surfaced microthreaded implants showed that implants with the microthreaded design caused minimal changes in crestal bone levels during healing (stress-free) and under functional loading.

Biofunctionalization of titanium implants with a biomimetic active peptide (P‐15) promotes early osseointegration

OBJECTIVES The early stages of peri-implant bone formation play an essential role in the osseointegration and long-term success of dental implants. By incorporating bioactive coatings, this biofunctionalization of implant surfaces may enhance the attachment of the implant to the surrounding bone and stimulate bone regeneration. MATERIAL AND METHODS To demonstrate faster osseointegration, the surfaces of dental implants were grit-blasted and acid-etched. They were then coated with hydroxyapatite (HA) and experimental implants were further coated with a biomimetic active peptide (P-15) in one of two concentrations. These biofunctionalized samples and controls with no peptide were placed in the forehead region of 12 adult pigs. Six animals were evaluated for a period of 14 or 30 days. RESULTS Histomorphometric analysis demonstrated that the implants with the high concentration of P-15 had significantly higher percentage of bone-to-implant contact (BIC) at 14 (P=0.018) and 30 (P=0.015) days compared with the other groups. Both concentrations of P-15 showed increased peri-implant bone density compared to the control group at 30 days. CONCLUSION Biofunctionalization of the implant surface with a biomimetic active peptide leads to significantly increased BIC rates at 14 and 30 days and higher peri-implant bone density at 30 days.

Effects of bioactive glass and beta-TCP containing three-dimensional laser sintered polyetheretherketone composites on osteoblasts in vitro

Because of their excellent physical properties nonresorbable thermoplastic polymers have become more important for the field of reconstructive surgery. Aim of the present study was to investigate the effects of laser sintered polyetheretherketone (PEEK) with incorporated osteoconductive and bioactive bone substitution materials on osteoblasts in vitro. Human osteoblasts (hFOB 1.19) were seeded onto laser sintered PEEK samples containing nano-sized carbon black, beta-tricalciumphosphate (beta-TCP), and bioactive glass 45S5. Osteoblasts were investigated for cell viability, cell proliferation and cell morphology. A constant proliferation of osteoblasts could be observed on all samples with the highest values for bioactive glass containing samples at day 7 (OD 1.76 +/- 0.22) and day 14 (OD 3.75 +/- 0.31) and lowest values for beta-TCP containing probes throughout the study compared with the PEEK pure control group. Highest cell viability was observed for Bioglass containing probes (95.5 +/- 3.32)% whereas osteoblasts seeded on beta-TCP containing probes showed reduced viability (84.4 +/- 4.32)%. Laser sintered PEEK implants seem to be attractive candidates for use as bone substitutes for reconstructive surgery because of their biocompatibility, individual shape, and the possibility of compounding bioinert polymer powder with osteoconductive and bioactive materials which might benefit bone formation in vivo.

Evaluation of the difference in accuracy between implant placement by virtual planning data and surgical guide templates versus the conventional free-hand method – a combined in vivo – in vitro technique using cone-beam CT (Part II)

PURPOSE The purpose of this study was to assess the accuracy of implant placement after virtual planning of implant positions using cone-beam CT data and surgical guide templates, and to match the results with those achieved with the conventional free-hand method. MATERIALS AND METHODS Twenty-three implants were placed in 10 patients with a Kennedy Class II with 3-dimensional (3-D) planned surgical guide template. Manual implantation was performed in anatomical casts of the same patients by a prosthodontist and a maxillofacial surgeon. Postoperative images of casts were superimposed onto the preoperative image of virtual planned ideal position of the implant. RESULTS The 3-D surgical guide template produced significantly smaller variation between the planned and actual implant positions at the implant shoulder (0.9 mm (0-4.5)) and apex (0.6-0.9 mm (0.0-3.4)) compared with the free-hand implantation (2.4-3.5 mm (0.0-7.0); p=0.000 and 2.0-2.5 mm (0.0-7.7); p=0.002). Accuracy of axis was also significantly improved. CONCLUSIONS Accuracy of implant placement after virtual planning of implant position using cone-beam CT data and surgical templates is high and significantly more accurate than free-hand insertion. The demonstrated method of superimposing radiographic images of postoperative casts and virtual planning images is a useful method, which allows reduced patient radiation exposure.

Bone regeneration after topical BMP-2-gene delivery in circumferential peri-implant bone defects.

OBJECTIVES The aims of this study were to evaluate the rate of bone formation and osseointegration after topical gene delivery with a liposomal vector system carrying bone morphogenetic protein (BMP)-2 cDNA in combination with a collagen carrier and autologous bone as a carrier in freshly created peri-implant bone defects. MATERIALS AND METHODS Eight domestic pigs received nine calvariae defects each (10 x 7 mm). A dental implant was inserted into the centre of each defect. In the test groups, the remaining space was filled with the liposomal vector/BMP-2 complex combined with a collagen carrier (n=18) or an autologous bone graft (n=18). Control groups were collagen only (n=18) and autologous bone graft only (n=18). RESULTS There was a significant difference in mineralisation rate in the BMP-2/bone graft (29.9%+/- 4.8 and 68.3%+/- 7.2) and bone graft only (22.6%+/- 2.6 and 49.4%+/- 13.9) groups after 7 and 28 days. Mineralisation values were also significantly higher in the BMP-2/collagen group (21.2%+/- 16.2 and 53.1%+/- 12.5) compared with the collagen-only group (8.2%+/- 7 and 41%+/- 8.1) in two different regions after 28 days. Also the bone-to-implant contact was significantly increased in the BMP-2/bone graft group after 28 days and in the BMP-2/collagen group after 7 and 28 days compared with their control groups. CONCLUSIONS The results of this study show a significantly positive effect of liposomal vector/BMP-2 on bone regeneration and osseointegration in bony circumferential peri-implant defects.

The diameter of anodic TiO2 nanotubes affects bone formation and correlates with the bone morphogenetic protein-2 expression in vivo

INTRODUCTION Recently, it has been demonstrated that the nanoscale environment is a critical factor for cellular behaviour. It has been shown that the diameter of TiO2 nanotube layers controls the cellular behaviour of cells involved in the bone-forming process in vitro. Therefore, the aim of the present study was to investigate the effects of the diameter of TiO2 nanotubes on peri-implant bone formation and the expression of bone matrix proteins in vivo. MATERIALS AND METHODS Ninety experimental implants with a nanotube diameter ranging from 15 up to 100 nm were placed in the frontal skulls of six domestic pigs, whereas untreated implants served as controls. The bone-implant contact (BIC) as well as the expression of bone morphogenetic protein (BMP)-2, collagen type-I and osteocalcin were histomorphometrically and immunohistochemically analysed after 30 days. RESULTS Evaluating the BIC, a significant higher value, could be found for the 50, 70 and 100 nm groups compared with the controls, whereas a correlation with the BMP-2 expression was present. The BMP-2 expression within the 50, 70 and 100 nm groups was statistically different compared with the control group. Significant difference was found for the osteocalcin expression in the 70 nm group. No statistical difference was found evaluating collagen type-I. SEM evaluation of the specimen surfaces revealed that the nanotube coatings do resist shearing forces evoked by implant insertion. CONCLUSION The nanotube diameter can be designed to support cellular functions of osteoblasts and osteoclasts in vivo, including differentiation and protein expression and therefore offer a powerful tool for the controlled formation of peri-implant bone around medical implant devices.

Vestibuloplasty: Porcine Collagen Matrix Versus Free Gingival Graft: A Clinical and Histologic Study

BACKGROUND A free gingival graft (FGG) is currently the gold standard for augmenting small areas of keratinized mucosa. The porcine collagen matrix (CM) represents an alternative to autologous tissue harvesting. This study aims to compare the CM versus FGGs for augmenting keratinized peri-implant mucosa based on clinical and histologic evaluations. METHODS The study included 14 patients who underwent a vestibuloplasty with either a FGG from the palate (n = 7) or the CM (n = 7). An implant-fixed vestibular retention splint was inserted for 30 days. Follow-up examinations were performed at 4, 10, 30, and 90 days after surgery. Width of keratinized mucosa was measured in the region of each implant (days 10, 30, and 90). After 90 days, a biopsy was harvested for histologic and immunohistologic analyses. To characterize newly formed soft tissue, the authors stained for tissue-and differentiation-specific markers, cytokeratin (CK) 5/6, 13, and 14, to detect presence or absence of keratinization. RESULTS The groups showed similar healing, with increased peri-implant keratinized mucosa. The CM group had overall significantly shorter operation times than the FGG group. Both groups showed similar overall shrinkage (32.98% CM versus 28.35% FGG). All biopsies showed a multilayered, keratinized, squamous epithelium. CKs 5/6 and 14 were detected in the basal and suprabasal layers, and spots of CK 13 were detected in the suprabasal layer. CONCLUSIONS During the whole observation period, both groups showed comparable clinical and histologic outcomes. Within the limitations of the present study, CM seems to be a promising alternative for the regeneration of keratinized mucosa without tissue harvesting. Comparative long-term studies are needed to investigate changes over time.