Bone regeneration in critical-size defects of the mandible using biomechanically adapted CAD/CAM hybrid scaffolds: An in vivo study in miniature pigs.

The study aimed to analyze bone regeneration in critical-size defects using hybrid scaffolds biomechanically adapted to the specific defect and adding the growth factor rhBMP-2. For this animal study, ten minipigs underwent bilateral defects in the corpus mandibulae and were subsequently treated with novel cylindrical hybrid scaffolds. These scaffolds were designed digitally to suit the biomechanical requirements of the mandibular defect, utilizing finite element analysis. The scaffolds comprised zirconium dioxide-tricalcium phosphate (ZrO2-TCP) support struts and TCP foam ceramics. One scaffold in each animal was loaded with rhBMP-2 (100 µg/cm³), while the other served as an unloaded negative control. Fluorescent dyes were administered every 2 weeks, and computed tomography (CT) scans were conducted every 4 weeks. Euthanasia was performed after 3 months, and samples were collected for examination using micro-CT and histological evaluation of both hard and soft tissue. Intravital CT examinations revealed minor changes in radiographic density from 4 to 12 weeks postoperatively. In the group treated with rhBMP-2, radiographic density shifted from 2513 ± 128 (mean ± SD) to 2606 ± 115 Hounsfield units (HU), while the group without rhBMP-2 showed a change from 2430 ± 131 to 2601 ± 67 HU. Prior to implantation, the radiological density of samples measured 1508 ± 30 mg HA/cm³, whereas post-mortem densities were 1346 ± 71 mg HA/cm³ in the rhBMP-2 group and 1282 ± 91 mg HA/cm³ in the control group (p = 0.045), as indicated by micro-CT measurements. The histological assessment demonstrated successful ossification in all specimens. The newly formed bone area proportion was significantly greater in the rhBMP-2 group (48 ± 10%) compared with the control group without rhBMP-2 (42 ± 9%, p = 0.03). The mean area proportion of remaining TCP foam was 23 ± 8% with rhBMP-2 and 24 ± 10% without rhBMP-2. Successful bone regeneration was accomplished by implanting hybrid scaffolds into critical-size mandibular defects. Loading these scaffolds with rhBMP-2 led to enhanced bone regeneration and a uniform distribution of new bone formation within the hybrid scaffolds. Further studies are required to determine the adaptability of hybrid scaffolds for larger and potentially segmental defects in the maxillofacial region.

Effect of enriched bone-marrow aspirates on the dimensional stability of cortico-cancellous iliac bone grafts in alveolar ridge augmentation.

BACKGROUND: The objective of the current study was to assess the clinical and radiological outcomes following autologous grafting from the iliac crest treated with autologous stem cells in-situ to reduce the postoperative bone graft resorption rate. MATERIALS AND METHODS: The study group consisted of patients who underwent vertical augmentation of the jaws via bone grafts harvested from the iliac crest enriched with bone-marrow aspirate concentrates (stem cell group). The first control group (control) included 40 patients underwent a vertical augmentation with autologous bone grafts from the iliac crest. In the second control group, 40 patients received identical surgical procedure, whereas the autologous bone graft was covered with a thin layer of deproteinized bovine bone matrix and a collagen membrane (DBBM group). Clinical complications, implant survival, radiological assessment of the stability of the vertical height and histological evaluation at the recipient site have been followed up for 24 months postoperatively. RESULTS: No differences in terms of implant survival were observed in the groups. In the stem cell group, the resorption after 4-6 months was 1.2 ± 1.3 mm and significantly lower than the resorption of the control group with 1.9 ± 1.6 mm (P = 0.029) (DBBM group: 1.4 ± 1.2 mm). After 12 months, the resorption of the stem cell group was 2.1 ± 1.6 mm and significantly lower compared to the control group (4.2 ± 3.0 mm, P = 0.001) and DBBM group (resorption 2.7 ± 0.9 mm, P = 0.012). The resorption rate in the second year was lower compared to the first year and was measured as 2.7 ± 1.7 mm in the stem cell group (1-year bone loss in the time period of 12-24 months of 0.6 mm compared to 2.1 mm in the first 12 months). The resorption was significantly lower compared to the control group (4.7 ± 2.9 mm; P = 0.003, DBBM group: 3.1 ± 0.5 mm, P = 0.075). CONCLUSIONS: Autologous bone-marrow aspirate concentrate could enhance the dimensional stability of the bone grafts and improve the clinical standard of complex reconstruction of the alveolar ridge. Even though the intraoperative cell enrichment requires an additional equipment and technical specification, it represents an alternative method for in-situ regeneration by osteogenic induction with a contribution of a manageable cost factor.

BMP-2 Long-Term Stimulation of Human Pre-Osteoblasts Induces Osteogenic Differentiation and Promotes Transdifferentiation and Bone Remodeling Processes.

Bone morphogenic protein (BMP-) 2 plays an important role in the regeneration of bone defects by promoting osteogenic differentiation. However, several animal studies have reported adverse side effects of BMP-2, including osteoclast activation, induction of peroxisome proliferator- activated receptor gamma (PPARG)expression, and inflammation. High BMP-2 concentrations are thought to be responsible for these side effects. For this reason, primary pre-osteoblasts were exposed to lower BMP-2 concentrations (1 and 2 µg/mL). Long-term exposure (up to 28 days) was performed to investigate whether this stimulation protocol may promote osteogenic differentiation without causing the side effects mentioned above. The results showed that BMP-2 treatment for 14 or 28 days resulted in increased osteogenesis, through an increase in runt-related transcription factor 2, osterix, alkaline phosphatase, and integrin-binding sialoprotein expression. However, an increase in tumor necrosis factor alpha and receptor activator of nuclear factor kappa-Β ligand protein levels was observed after BMP-2 exposure, indicating also an increased potential for osteoclast activation by osteoblasts. Additionally, morphological changes like intracellular, filled vacuoles could be detected. Enhanced PPARG and perilipin 1 mRNA transcripts and lipid droplets indicated an induced adipogenic differentiation. Overall, the data demonstrate that long-term BMP-2 exposure promotes not only osteogenic differentiation but also adipogenesis and regulates mediators involved in osteoclast activation in vitro.

In vivo biocompatibility evaluation of 3D-printed nickel-titanium fabricated by selective laser melting.

Nickel-titanium (NiTi) belongs to the group of shape-memory alloys (SMAs), which are characterized by flexibility and reversible deformability. Advanced techniques in 3D printing by selective laser-melting (SLM) process allow the manufacturing of complex patient-specific implants from SMAs. Osteosynthesis materials made of NiTi could be used for minimally invasive surgical approaches in oral- and maxillofacial surgery. However, the in vivo biocompatibility has not yet been fully investigated, especially in load-sharing and load-bearing implants. The aim of this study was to evaluate the in vivo biocompatibility of SLM-produced NiTi for intraosseous and subperiosteal applications. Test specimens were implanted into the frontonasal bone of ten miniature pigs. To assess peri-implant bone metabolism, fluorescent dye was administered after 2, 4, 6, 10, 12, and 14 weeks intraperitoneally. Specimens and the surrounding tissues were harvested after 8 and 16 weeks for histological analysis. While the NiTi implants presented a higher bone-to-implant contact ratio (BIC) after 8 than after 16 weeks (43.3 vs. 40.3%), the titanium implants had a significantly higher BIC after 16 weeks (33.6 vs. 67.7%). Histologically, no signs of peri-implant inflammation or foreign-body reaction were detectable. With respect to this preliminary study design, 3D-printed NiTi shows sufficient biocompatibility for intraosseous and subperiosteal implant placement.

Therapy of antigen-induced arthritis of the temporomandibular joint via platelet-rich plasma injections in domestic pigs.

The aim of this animal study was to investigate the effect of platelet-rich plasma (PRP) injections into the temporomandibular joint (TMJ) to treat antigen-induced arthritis AIA. AIA was induced via the application of bovine serum albumin (BSA) into the TMJ. Clear histological signs and protein analysis results indicating inflammation of the TMJ were observed. Afterwards, two PRP injections were performed over an interval of 2 weeks. Concentration levels of the proinflammatory cytokines IL-1ß (PRP: 33.7 ± 5.6 pg/mg, untreated: 50.0 ± 2.9 pg/mg; p = 0.04) and TNF-α (PRP: 20.7 ± 2.5 pg/mg, untreated: 31.4 ± 2.7 pg/mg; p = 0.03) were significantly decreased in the PRP-treated joints. A significant reduction in signs of histological inflammation, such as hyperplasia of the synovial membrane, leucocyte infiltration, cartilage surface alterations, and an increase in cartilage-specific glycosaminoglycan content, was observed. This animal study supports the understanding of the underlying effects of PRP treatment in the TMJ, and may enhance novel PRP therapies in the future.

Bone tissue engineering in the greater omentum with computer-aided design/computer-aided manufacturing scaffolds is enhanced by a periosteum transplant.

Aim: This study aimed to evaluate two different vascularized bone flap scaffolds and the impact of two barrier membranes for the reconstruction of critical-size bone defects. Materials & methods: 3D-printed scaffolds of biodegradable calcium phosphate and bioinert titanium were loaded with rhBMP-2 bone marrow aspirate, wrapped by a collagen membrane or a periosteum transplant and implanted into the greater omentum of miniature pigs. Results: Histological evaluation demonstrated significant bone formation within the first 8 weeks in both scaffolds. The periosteum transplant led to enhanced bone formation and a homogenous distribution in the scaffolds. The omentum tissue grew out a robust vascular supply. Conclusion: Endocultivation using 3D-printed scaffolds in the greater omentum is a very promising approach in defect-specific bone tissue regeneration.

Transcatheter mitral valve implantation: supra-annular and subvalvular fixation techniques.

OBJECTIVES: Transcatheter mitral valved stent implantation provides an off-pump treatment option for mitral valve regurgitation, especially for secondary mitral valve regurgitation. The aim of this study was to evaluate novel fixation strategies: direct fixation (SUPRA) and subvalvular fixation (sub-VALV) to successfully implement alternative fixation methods at the mitral annulus and to reduce radial stent and apical tether forces. METHODS: Specific concepts were developed for the supra-annular hook-shaped fixation (SUPRA) and the subvalvular fixation (sub-VALV). These prototypes were compared with the sole apical tether fixation (AP) methods. Thirty-three pigs underwent mitral valved stent implantation accompanied by standardized transoesophageal echocardiographic and haemodynamic evaluation of heart function and the stent performance 1 h after implantation. Additionally, animals were followed up for 3 months. RESULTS: Secure deployment and correct positioning with low transvalvular gradients were achieved in all cases with mitral valved stent implantation. Nevertheless, 2 pigs died due to rhythm disturbances during dissection and pre-transcatheter mitral valve implantation. Paravalvular leakages were trace or less in prototypes with supra-annular fixation and sole apical fixation. In contrast, moderate paravalvular leakages were observed in the sub-VALV group (P < 0.001). In addition, the effect of a specific stent design on heart function was demonstrated: an increased rate of ischaemia and arrhythmia (P = 0.037) and a small left ventricular ejection fraction reduction (P < 0.05) were observed in the group with subvalvular fixation. In all but 1 case, gross evaluation demonstrated a good ingrowth of the valved stents of between 36% and 100% tissue coverage after a follow-up of 1 month or longer. CONCLUSIONS: The low degree of paravalvular leakages in the supra-annular and apical fixation groups and the normal longitudinal function highlight the good alignment of these new mitral valved stent designs within the native anatomy. The novel, hook-shaped, supra-annular fixation elements were well tolerated in the hearts resulting in excellent health of the animals at long-term follow-up times of up to 3 months.

Novel stent design for transcatheter mitral valve implantation.

OBJECTIVES: In this study, results of a functional in vitro study of 2 newly developed valved stents for transcatheter mitral valve implantation are presented. METHODS: Two novel stent designs, an oval-shaped and a D-shaped stent with a strut fixation system were developed. The fixation force of the novel stents were tested in vitro in porcine hearts with a tensile test set-up. In further experiments, the stents were equipped with a circular valved stent, and the valve performances were investigated in a pulsatile heart valve tester. RESULTS: Sufficient mean stent fixation forces in the range of 24.2 ± 0.9 N to 28.6 ± 1.9 N were measured for the different stent models. The novel valved stents showed good performance in an in vitro pulsatile heart valve tester. A sufficient opening area and low opening pressures were measured for all tested mitral valved stents. Compared with an established reference valve, the D-shaped stent and the oval-shaped valved stent showed a lower systolic transvalvular pressure gradient, which indicates slightly greater extent of valvular leakage of the closed valved stents. However, the mitral nitinol valved stents demonstrated adequate durability. CONCLUSIONS: This study indicates a sufficient annular fixation force of the tested transcatheter mitral valve implantation valved stent prototypes. Therefore, these mitral valved stents demonstrate a new type of mitral valved stent design.