title: Biotechnological approach for systemic delivery of membrane Receptor Activator of NF-κB Ligand (RANKL) active domain into the circulation.
abstract: Deficiency of Receptor Activator of NF-κB Ligand (RANKL) prevents osteoclast formation causing osteopetrosis. RANKL is a membrane-bound protein cleaved into active soluble (s)RANKL by metalloproteinase 14 (MMP14). We created a bio-device that harbors primary osteoblasts, cultured on 3D hydroxyapatite scaffolds carrying immobilized MMP14 catalytic domain. Scaffolds were sealed in diffusion chambers and implanted in RANKL-deficient mice. Mice received 1 or 2 diffusion chambers, once or twice and were sacrificed after 1 or 2 months from implants. A progressive increase of body weight was observed in the implanted groups. Histological sections of tibias of non-implanted mice were negative for the osteoclast marker Tartrate-Resistant Acid Phosphatase (TRAcP), consistent with the lack of osteoclasts. In contrast, tibias excised from implanted mice showed TRAcP-positive cells in the bone marrow and on the bone surface, these latter morphologically similar to mature osteoclasts. In mice implanted with 4 diffusion chambers total, we noted the highest number and size of TRAcP-positive cells, with quantifiable eroded bone surface and significant reduction of trabecular bone volume. These data demonstrate that our bio-device delivers effective sRANKL, inducing osteoclastogenesis in RANKL-deficient mice, supporting the feasibility of an innovative experimental strategy to treat systemic cytokine deficiencies.
authors: Regenerative Medicine Unit, Ospedale Pediatrico Bambino Gesù Istituto di Ricovero e Cura a Carattere Scientifico, Piazza S. Onofrio 4, 00165 Rome, Italy.
authors: Alfredo
authors: A
authors: Cappariello
authors: Department of Biotechnological and Applied Clinical Sciences, University of L'Aquila, Via Vetoio - Coppito 2, 67100 L'Aquila, Italy.
authors: Riccardo
authors: R
authors: Paone
authors: Department of Biotechnological and Applied Clinical Sciences, University of L'Aquila, Via Vetoio - Coppito 2, 67100 L'Aquila, Italy.
authors: Antonio
authors: A
authors: Maurizi
authors: Department of Biotechnological and Applied Clinical Sciences, University of L'Aquila, Via Vetoio - Coppito 2, 67100 L'Aquila, Italy.
authors: Mattia
authors: M
authors: Capulli
authors: Department of Biotechnological and Applied Clinical Sciences, University of L'Aquila, Via Vetoio - Coppito 2, 67100 L'Aquila, Italy.
authors: Nadia
authors: N
authors: Rucci
authors: Regenerative Medicine Unit, Ospedale Pediatrico Bambino Gesù Istituto di Ricovero e Cura a Carattere Scientifico, Piazza S. Onofrio 4, 00165 Rome, Italy.
authors: Maurizio
authors: M
authors: Muraca
authors: Department of Biotechnological and Applied Clinical Sciences, University of L'Aquila, Via Vetoio - Coppito 2, 67100 L'Aquila, Italy. Electronic address: annamaria.teti@univaq.it.
authors: Anna
authors: A
authors: Teti
mesh_terms: D000818:Animals; D001709:Biotechnology / Q000379:methods*; D001862:Bone Resorption / Q000473:pathology; D002449:Cell Aggregation / Q000187:drug effects; D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000494:pharmacology; D016503:Drug Delivery Systems; D017886:Durapatite / Q000494:pharmacology; D006801:Humans; D020007:Implants, Experimental; D008168:Lung / Q000473:pathology; D008214:Lymphocytes / Q000187:drug effects / Q000473:pathology; D053511:Matrix Metalloproteinase 14 / Q000378:metabolism; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010006:Osteoblasts / Q000187:drug effects / Q000378:metabolism; D010010:Osteoclasts / Q000187:drug effects / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects; D017434:Protein Structure, Tertiary; D053245:RANK Ligand / Q000097:blood* / Q000737:chemistry*; D012995:Solubility
chemical_list: D017077:Culture Media, Conditioned; D053245:RANK Ligand; D017886:Durapatite; D053511:Matrix Metalloproteinase 14
keywords: Diffusion chambers; Osteoclast; Osteopetrosis; RANKL; Receptor Activator of NF-κB Ligand