Many studies have highlighted the role of silicon in human bone formation and maintenance. Silicon, in fact, is considered to nucleate the precipitation of hydroxyapatite and to reduce the bone resorption. For this reason, we have combined silk fibroin (SF) with silicon-releasing diatom particles (DPs), as potential material for bone tissue engineering applications. Sponges of fibroin loaded with different amounts and sizes of DPs were prepared by solvent casting-particulate leaching method, and their morphology, porosity and mechanical properties were evaluated. The biological effect of diatom addition was assessed on human osteosarcoma cell line MG63, a suitable osteoblast-like model, through cell adhesion, metabolic activity and proliferation assays. In addition, alkaline phosphatase activity, osterix and collagen type I production in MG63 cell line were assessed as markers of early bone formation to demonstrate a pro-mineralization potential of scaffolds. Results of the studies showed that addition to fibroin of diatoms particles improved the osteogenic properties of osteoblast-like cells compared with the pure SF.