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One property that can be added is the production\/release of bioactive soluble factors, either directly from the biomaterial, or from cells embedded within the biomaterial. We reasoned that pursuing this strategy would be appropriate to setup a cell-based therapy for RANKL-deficient autosomal recessive osteopetrosis, a very rare skeletal genetic disease in which lack of the essential osteoclastogenic factor RANKL impedes osteoclast formation. The exogenously administered RANKL cytokine is effective in achieving osteoclast formation and function in vitro and in vivo, thus, we produced murine Rankl\u2212\/\u2212 mesenchymal stromal cells (MSCs) overexpressing human soluble RANKL (hsRL) following lentiviral transduction (LVhsRL). Here, we described a three-dimensional (3D) culture system based on a magnesium-doped hydroxyapatite\/collagen I (MgHA\/Col) biocompatible scaffold closely reproducing bone physicochemical properties. MgHA\/Col-seeded murine MSCs showed improved properties, as compared to two-dimensional (2D) culture, in terms of proliferation and hsRL production, with respect to LVhsRL-transduced cells. When implanted subcutaneously in Rankl\u2212\/\u2212 mice, these cell constructs were well tolerated, colonized by host cells, and intensely vascularized. Of note, in the bone of Rankl\u2212\/\u2212 mice that carried scaffolds with either WT or LVhsRL-transduced Rankl\u2212\/\u2212 MSCs, we specifically observed formation of TRAP+ cells, likely due to sRL released from the scaffolds into circulation. Thus, our strategy proved to have the potential to elicit an effect on the bone; further work is required to maximize these benefits and achieve improvements of the skeletal pathology in the treated Rankl\u2212\/\u2212 mice. Stem Cells Translational Medicine \u00a02019;8:22\u201334<\/jats:p>","DOI":"10.1002\/sctm.18-0085","type":"journal-article","created":{"date-parts":[[2018,9,5]],"date-time":"2018-09-05T16:42:10Z","timestamp":1536165730000},"page":"22-34","source":"Crossref","is-referenced-by-count":40,"title":["Mesenchymal Stromal Cell-Seeded Biomimetic Scaffolds as a Factory of Soluble RANKL in Rankl-Deficient Osteopetrosis"],"prefix":"10.1093","volume":"8","author":[{"given":"Ciro","family":"Menale","sequence":"first","affiliation":[{"name":"a CNR-IRGB, Milan Unit, Milan, Italy"},{"name":"b Humanitas Clinical and Research Institute, Rozzano, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elisabetta","family":"Campodoni","sequence":"additional","affiliation":[{"name":"c CNR-ISTEC, Faenza, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eleonora","family":"Palagano","sequence":"additional","affiliation":[{"name":"b Humanitas Clinical and Research 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Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elena","family":"Fontana","sequence":"additional","affiliation":[{"name":"a CNR-IRGB, Milan Unit, Milan, Italy"},{"name":"b Humanitas Clinical and Research Institute, Rozzano, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Francesca","family":"Schena","sequence":"additional","affiliation":[{"name":"e Clinica Pediatrica e Reumatologia UOSD Centro Malattie Autoinfiammatorie e Immunodeficienze, Genoa, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rob","family":"van\u2019t Hof","sequence":"additional","affiliation":[{"name":"f Bone Research Group, Institute of Ageing & Chronic Disease University of Liverpool, Liverpool, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Monica","family":"Sandri","sequence":"additional","affiliation":[{"name":"c CNR-ISTEC, Faenza, 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