In vitro proliferation and differentiation of human mesenchymal stem cells on hydroxyapatite versus human demineralised bone matrix with and without osteogenic protein-1

Abstract

Background: Allografting is currently used in lower limb reconstruction surgery. Demineralised bone matrix (DBM) is more osteoinductive compared with allografts but lacks mechanical strength. Osteogenic protein-1 (OP-1) can improve the osteoinductivity of the allograft, however recent reports indicate significant allograft resorption when it is combined with OP-1. Objectives: Our hypothesis was that hydroxyapatite (HA) with human-mesenchymal stem cells (h-MSCs) and OP-1 (HA+h-MSCs+OP-1) has similar osteoinductive properties to human-DBM+h-MSCs. The objective was to evaluate h-MSC proliferation (by tritiated thymidine incorporation, total DNA Hoechst 33258 and scanning electron microscopy) and osteogenic differentiation (from alkaline phosphatase activity) in human demineralised bone matrix (h-DBM) and HA, with or without OP-1. Results: H-MSC proliferation on HA+OP-1 was significantly higher compared with that on HA at all time points (p < 0.05) and compared with DBM alone [day 1, (198.4 vs 95.4) p = 0.042; day 14 (286.1 vs 119.9), p < 0.001]. H-MSC proliferation was higher in DBM+OP-1, at all time points compared with HA+OP-1 but the difference was not statistically significant (p > 0.05). H-MSC differentiation was significantly higher in HA+OP-1 compared with HA (p < 0.05) but not significantly different from diffferentiation on DBM alone (p > 0.05). Differentiation was significantly higher on DBM+OP-1 at all time points compared with HA (p < 0.05) and with HA+OP-1 [day 1, (21.1 vs 10.1) (p = 0.03); day 7 (39.4 vs 7.1) (p < 0.01); day 14 (40.2 vs 14.4) (p < 0.001)]. Conclusions: When HA+h-MSCs is combined with OP-1 in vitro its osteogenic potential is similar to that of DBM+h-MSCs alone which may be adequate for non-weight-bearing applications. Mechanical testing however is of great importance for weight-bearing applications and the in vivo testing of the composite graft HA+h-MSCs+OP-1 vs DBM+h-MSCs is recommended.