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.

Osseointegration of dental implants in ectopic engineered bone in three different scaffold materials.

The in vivo regeneration of bone flaps might be an alternative to autogenous bone grafting. The first human case of mandibular reconstruction using the greater omentum as a bioreactor was reported in 2016. However, whether engineered bone will support the osseointegration of dental implants has not yet been investigated. In this study, bone tissue engineering was performed in the greater omentum of nine miniature pigs using bone morphogenetic protein 2, bone marrow aspirate, and three different scaffolds: hydroxyapatite, biphasic calcium phosphate (BCP), and titanium. After 8 weeks, two implants were placed in each scaffold; after another 8 weeks, the bone blocks were harvested for radiographic, histological, and histomorphometric analysis. All implants exhibited sufficient primary stability, and the success rate was 100%. The bone-to-implant contact ratios (BICs) were 38.2%, 68.5%, and 42.9%; the inter-thread bone densities were 29.4%, 64.9%, and 33.5%; and the peri-implant bone-scaffold densities were 56.4%, 87.6%, and 68.6% in the hydroxyapatite, BCP, and titanium groups, respectively. The BIC showed a strong correlation (r = 0.76) with the peri-implant bone-scaffold density. This study shows that de novo engineered bone leads to successful osseointegration and therefore may allow implant-based prosthodontic rehabilitation.

Bone tissue engineering in the greater omentum is enhanced by a periosteal transplant in a miniature pig model.

AIM: Reconstruction of bone defects with autologous grafts has certain disadvantages. The aim of this study is to introduce a new type of living bioreactor for engineering of bone flaps and to evaluate the effect of different barrier membranes. MATERIALS & METHODS: Scaffolds loaded with bone morphogenetic proteins and bone marrow aspirate wrapped with either a collagen membrane or a periosteal flap were implanted in the greater omentum of miniature pigs. RESULTS: Both histological and radiographic evaluation showed proven bone formation and increased density after 8 and 16 weeks, with an enhanced effect of the periosteal transplant. CONCLUSION: The greater omentum is a suitable bioreactor for bone tissue engineering. Endocultivation is both an innovative and promising approach in regenerative medicine.

* Endocultivation of Scaffolds with Recombinant Human Bone Morphogenetic Protein-2 and VEGF165 in the Omentum Majus in a Rabbit Model.

The reconstruction of defects in the mandible are still challenging. Despite several adequate microvascular bone reconstruction techniques, there is a need for ectopic bone endocultivation without drawbacks by donor-site morbidity. The omentum majus is described as a good vascularized fleece with undifferentiated cells with potential for bone culturing. In the omentum majus of six rabbits, two hydroxyapatite blocks were incorporated for 12 weeks each. The blocks were prepared with recombinant human bone morphogenetic protein-2 (rhBMP-2) or VEGF165 + rhBMP-2 and wrapped into the omentum. For ectopic bone endocultivation observation computed tomography (CT) scans were performed, and fluorescence markers were applied. After harvesting the block, histological sections were performed with hematoxylin and eosin and toluidine blue staining. In the CT scans, the Hounsfield units of the blocks increased within the trail. In some sections, new bone formation was observed within the hydroxyapatite blocks, however, the histological staining showed soft-tissue invasion only, no gross bone formation was observed. The ectopic bone endocultivation in the omentum majus is technically a good approach. An adequate mixture of osteoinductive proteins is still missing.

Endocultivation: continuous application of rhBMP-2 via mini-osmotic pumps to induce bone formation at extraskeletal sites.

The continuous presence of recombinant human bone morphogenetic protein 2 (rhBMP-2) inside a scaffold may be crucial to the outcome in bone tissue engineering. This study investigated whether the release of the growth factor rhBMP-2 via different continuous application schemes influences histomorphological aspects of the hard and soft tissues induced. Three-dimensionally printed hydroxyapatite scaffolds were implanted into one latissimus dorsi muscle of 42 female Lewis rats. Simultaneously implanted mini-osmotic pumps were used to provide a continuous application of rhBMP-2 over 1, 2, or 4 weeks (total dose 200µg). A reference group received rhBMP-2 at the time of implantation only, and a control group received only block implantation. Bone density and histological examinations were performed after 8 weeks. No significant difference in bone density was found between the groups; however, the blood vessel count differed significantly between the groups receiving continuous treatments and both the control group and simultaneous rhBMP-2 treatment group (P<0.0001). Soft tissue types were distributed differently among the study groups. RhBMP-2 application via mini-osmotic pumps is as suitable for inducing bone formation as a single application at the time of implantation. The time interval over which rhBMP-2 was administered had no impact on the amount of new bone formation, probably due to the study duration and low local concentrations of growth factor.

Endocultivation: Histomorphological effects of repetitive rhBMP-2 application into prefabricated hydroxyapatite scaffolds at extraskeletal sites.

The timing of application of recombinant human bone morphogenic protein-2 (rhBMP-2) may be important in determining the final outcome of engineered bone tissue. This study investigates the impact of repetitive rhBMP-2 application on hard and soft tissue morphology in endocultivation. A 3D-printed scaffold was implanted into a pouch in the latissimus dorsi muscle in 40 Lewis rats. RhBMP-2 was injected at defined time points and animals received a total of 200 µg each. Control groups received either rhBMP-2 simultaneously with scaffold implantation, or solely a scaffold with no rhBMP-2. Fluorescence markers were injected after operation. CT-scans and histological examination were performed after 8 weeks. Multiple comparisons revealed significant differences of bone density between the groups who received delayed injections at two separate time points in time compared to those who had simultaneous rhBMP-2 application (p = 0.0038; p = 0.0063) and the control group (p = 0.017, p = 0.0284). The blood vessel count was significantly higher in groups with repetitive injections compared with both control groups. Two soft tissue types were identified and found to have different distributions in the different study groups. Fluorescence labeling showed active new bone formation after 4-5 weeks in all groups where rhBMP-2 was administered. Multiple repetitive injections were more effective than simultaneous application regarding bone density indicating time-dependent effects of rhBMP-2. Bone formation processes were detectable several weeks after rhBMP-2 application indicating long-term effects.