A Xenogenic Bone Derivative as a Potential Adjuvant for Bone Regeneration and Implant Osseointegration: An In Vitro Study

Several clinical conditions may limit the success of bone regeneration and/or implant osseointegration. For this reason, many compounds have been tested for their ability to stimulate this biological process. Synthetic hydroxyapatite (HA), mimicking natural bone hydroxyapatite, and extra-cellular matrix proteins, such as type I collagen, are potential candidates. However, the synthetic origin of HA and the denaturing conditions required for extracting collagen from skin and derma are sources of potential drawbacks. This study examines the in vitro effects of a natural bone derivative (NBD) extracted from equine bone and containing both natural, non-synthetic bone hydroxyapatite and native, non-denatured, type I bone collagen as a possible active compound for stimulating bone regeneration and implant osseointegration. The activity of NBD was tested on bone marrow stromal cells (BMSCs), evaluating their growth/viability by the methylthiazol tetrazolium (MTT) assay and their migration potential by a scratch assay. Moreover, expression of the hyaluronic acid receptor (CD44) and the C-X-C chemokine receptor type 4 (CXCR4, CD184) on the surface of BMSCs was assessed by flow cytometry, and the release of Transforming Growth Factor (TGF)-beta, Interleukin (IL)-1 alpha and IL-6 was quantified using an enzyme-linked immunosorbent assay (ELISA). The effect of NBD-coated implants on human osteoblasts was tested by measuring alkaline phosphatase (ALP) activity with the p-nitrophenyl phosphate (pNPP) degradation test. NBD stimulated BMSC growth/viability, migration, CD184 surface expression and the release of TGF-beta 1. NBD-coated implants increased ALP activity of human osteoblasts. These results indicate that NBD may be an adjuvant to accelerate both bone regeneration and osseointegration.