In the present study, we evaluated an autologous bone graft substitute (ABGS) composed of recombinant human BMP6 (rhBMP6) dispersed within autologous blood coagulum (ABC) used as a physiological carrier for new bone formation in spine fusion sheep models. The application of ABGS included cervical cage for use in the anterior lumbar interbody fusion (ALIF), while for the posterolateral lumbar fusion (PLF) sheep model allograft devitalized bone particles (ALLO) were applied with and without use of instrumentation. In the ALIF model, ABGS (rhBMP6/ABC/cage) implants fused significantly when placed in between the denuded L4-L5 vertebrae as compared to control (ABC/cage) which appears to have a fibrocartilaginous gap, as examined by histology and micro CT analysis at 16 weeks following surgery. In the PLF model, ABGS implants with or without ALLO showed a complete fusion when placed ectopically in the gutter bilaterally between two decorticated L4-L5 transverse processes at a success rate of 88% without instrumentation and at 80% with instrumentation; however the bone volume was 50% lower in the instrumentation group than without, as examined by histology, radiographs, micro CT analyses and biomechanical testing at 27 weeks following surgery. The newly formed bone was uniform within ABGS implants resulting in a biomechanically competent and histologically qualified fusion with an optimum dose in the range of 100 μg rhBMP6 per mL ABC, while in the implants that contained ALLO, the mineralized bone particles were substituted by the newly formed remodeling bone via creeping substitution. These findings demonstrate for the first time that ABGS (rhBMP6/ABC) without and with ALLO particles induced a robust bone formation with a successful fusion in sheep models of ALIF and PLF, and that autologous blood coagulum (ABC) serves as a preferred physiological native carrier to induce new bone at low doses of rhBMP6 and to achieve a successful spinal fusion.

Segmental defects of long bones are among the most challenging and debilitating conditions in clinical medicine. Osteogrow-C is a novel osteoinductive device composed of recombinant human bone morphogenetic protein 6 (rhBMP6) delivered within autologous blood coagulum (ABC) with calcium phosphate ceramics that was proven efficacious in preclinical models of spinal fusion. This study aimed to evaluate the efficacy of Osteogrow-C in comparison to that of other osteoinductive therapies in a clinically relevant segmental defect model in rabbits. Segmental defects (15 mm) of rabbit ulna were treated with Osteogrow-C containing different synthetic ceramic particles (tricalcium phosphate [TCP] and TCP/hydroxyapatite 40%/60%), Osteogrow (rhBMP6/ABC), Infuse (rhBMP2/absorbable collagen sponge), and control implants without bone morphogenetic proteins. Defect healing was evaluated by in vivo x-ray scans 4, 8, and 17 weeks after the surgery, and animals were killed after 17 weeks for further radiographical and histological assessment. Evaluation of x-ray images, micro-computed tomography, and histological sections revealed that both Osteogrow-C formulations as well as Osteogrow and Infuse promoted healing of the ulnar segmental defect. However, radiographic scores were higher in animals treated with Osteogrow-C than those for the other used therapies. Moreover, evaluation of in vivo x-ray scans revealed that Osteogrow-C with TCP ceramics induced the most rapid defect bridging. On the other hand, control implants (ABC/TCP and ABC/biphasic calcium phosphate) promoted limited osteogenesis without defect bridging. The findings of this study suggest that Osteogrow-C is a promising safe therapeutic solution for the treatment of large bone defects, providing relief to millions of patients suffering from this debilitating condition.

Extracellular matrix (ECM) has been intensively used in cardio surgery. The main goal of this research was to determine if Achilles tendon healing could be promoted by applying extracellular matrix scaffold (CorMatrix®, USA). Sixteen (n = 16) New Zealand white mature rabbits (Oryctolagus cuniculus) were randomly allocated into two groups. Following complete surgical transection, rabbits in group A (ECM applied) (n = 8) had their Achilles tendons reconstructed using both, nylon suture and extracellular matrix scaffold, whereas in group B (without ECM) the tendons were reconstructed using nylon suture only. After four weeks, the rabbits were euthanized and tendon samples harvested and stained with hematoxylin eosin, Mallory, and Gomory and subsequently histologically analyzed according to modified Bonnar scale. Group B had significantly stronger inflammatory response, including abundant cell infiltration and neovascularization. In group A collagen fibers were predominantly found, whereas in group B reticular fibers were more abundant. Extracellular matrix scaffold has been found to have the real potential for promoting tendon healing through accelerating collagen formation, which is crucial for restoring biomechanical properties of a tendon, decreasing peritendineous adhesion formation, and reducing inflammatory edema and subsequently pain.