Institut de Recherche Expérimentale et Clinique, Neuro Musculo-Skeletal Lab, Université Catholique de Louvain, Avenue E. Mounier, 52-B1.52.04 - 1200, Bruxelles, Belgium; Institut de Recherche Expérimentale et Clinique, Pôle Chirurgie Expérimentale et Transplantation, Université Catholique de Louvain, Avenue E. Mounier, 52-B1.52.04 - 1200, Bruxelles, Belgium; Service de Chirurgie Orthopédique et Traumatologique, Cliniques Universitaires Saint-Luc, Avenue Hippocrate 10-1200, Bruxelles, Belgium; Unité de Thérapie Tissulaire et Cellulaire de l'Appareil Locomoteur, Cliniques Universitaires Saint-Luc, Avenue Hippocrate 10-1200, Bruxelles, Belgium.
Institut de Recherche Expérimentale et Clinique, Neuro Musculo-Skeletal Lab, Université Catholique de Louvain, Avenue E. Mounier, 52-B1.52.04 - 1200, Bruxelles, Belgium; Service de Chirurgie Orthopédique et Traumatologique, Cliniques Universitaires Saint-Luc, Avenue Hippocrate 10-1200, Bruxelles, Belgium; Institut de Recherche Expérimentale et Clinique, Pôle Morphologie, Université Catholique de Louvain, Avenue E. Mounier, 52-B1.52.04 - 1200, Bruxelles, Belgium; Unité de Thérapie Tissulaire et Cellulaire de l'Appareil Locomoteur, Cliniques Universitaires Saint-Luc, Avenue Hippocrate 10-1200, Bruxelles, Belgium.
Bone. 2024 Oct;187:117213. doi: 10.1016/j.bone.2024.117213. Epub 2024 Jul 29.
Critical bone loss can have several origins: infections, tumors or trauma. Therefore, massive bone allograft can be a solution for limb salvage. Such a biological reconstruction should have the ideal biomechanical qualities. However, their complication rate remains too high. Perfusion-decellularization of massive allografts could promote the vitality of these grafts, thereby improving their integration and bone remodeling. Three perfusion-decellularized massive bone allografts were compared to 3 fresh frozen massive bone allografts in a preclinical in vivo porcine study using an orthopedic surgery model. Three pigs each underwent a critical diaphyseal femoral defects followed by an allogeneic intercalary femoral graft on their both femurs (one decellularized and one conventional fresh frozen as "native") to reconstruct the defect. Clinical imaging was performed over 3 months of follow-up. The grafts were then explanted and examined by non-decalcified histology, fluoroscopic microscopy and immunohistochemistry. Bone consolidation was achieved in both groups at the same time. However, the volume of bone callus appeared to be greater in the decellularized group. Histology demonstrated a superior bone remodeling in the decellularized group, with a higher number of osteoclasts (p < 0.001) and larger areas of osteoid matrix and newly formed bone as compared to the "native" group. Immunohistochemistry showed a superior vitality and remodeling in both the cortical and medullary cavities for osteocalcin (p < 0.001), Ki67 (p < 0.001), CD3 (p < 0.001) and α-SMA (p < 0.001) as compared the "native" group. Three months after implantation, the decellularized grafts were proven to be biologically more active compared to native grafts. Fluoroscopic microscopy revealed more ossification fronts in the depth of the decellularized grafts (p = 0.021). This pilot study provides the first in vivo demonstration on the enhanced biological capacities of massive bone allograft decellularized by perfusion as compared to conventional massive bone allografts.
感染、肿瘤或外伤。因此，大量同种异体骨移植可以作为保肢的一种解决方案。这种生物重建应该具有理想的生物力学特性。然而，其并发症发生率仍然过高。大量同种异体骨的灌注去细胞化可以促进这些移植物的活力，从而改善其整合和骨重塑。在一项使用骨科手术模型的临床前猪体内研究中，将 3 例灌注去细胞化的同种异体大块骨移植与 3 例新鲜冷冻的同种异体大块骨移植进行了比较。每只猪的双侧股骨均接受了临界骨干缺损，然后接受同种异体间插股骨移植（1 例去细胞化，1 例常规新鲜冷冻，作为“天然”）以重建缺损。在 3 个月的随访过程中进行临床影像学检查。然后将移植物取出，通过非脱钙组织学、荧光显微镜和免疫组织化学检查。两组均在同一时间实现了骨愈合。然而，去细胞化组的骨痂体积似乎更大。组织学显示，去细胞化组的骨重塑更好，破骨细胞数量更多（p<0.001），类骨质基质和新形成的骨面积也大于“天然”组。免疫组织化学显示，与“天然”组相比，骨钙素（p<0.001）、Ki67（p<0.001）、CD3（p<0.001）和α-SMA（p<0.001）在皮质和髓腔中均具有更高的活力和重塑。植入 3 个月后，与天然移植物相比，去细胞化移植物被证明具有更高的生物活性。荧光显微镜显示，去细胞化移植物深部有更多的骨化前沿（p=0.021）。这项初步研究首次在体内证明了灌注去细胞化的同种异体大块骨移植与传统同种异体大块骨移植相比，具有增强的生物学能力。
