Platelet-derived growth factor inhibits demineralized bone matrix-induced intramuscular cartilage and bone formation: A study of immunocompromised mice

Background: Platelet-derived growth factor (PDGF) has been proposed as a therapeutic agent to promote bone-healing. The purpose of this study was to examine the effect of PDGF on the ability of human demineralized bone matrix to induce bone formation in a nude-mouse muscle-implantation model. We also examined whether platelet-rich plasma, which contains PDGF, also modulates osteoinduction in this model. Methods: Human demineralized bone matrix, previously shown to be osteoinductive in the calf muscles of nude mice, was mixed with PDGF-BB (0, 0.1, 1, and 10 μg/10 mg of demineralized bone matrix) and was implanted bilaterally in the calf muscles of immunocompromised (nu/nu) mice (six mice in each group). Heat-inactivated demineralized bone matrix was used as a control. Tissue was harvested at fourteen, twenty-eight, and fifty-six days after implantation. Platelet-rich plasma was prepared from the blood of a healthy donor with use of the Harvest PRP preparation device, activated with thrombin, and mixed with active and inactive demineralized bone matrix. Fifty-six days post-implantation, tissues were harvested. Osteoinduction was assessed with use of a qualitative scoring system and with quantitative histomorphometry. Results: Cartilage was present at fourteen days in all tissues that had received an implant, but the amount decreased as the PDGF concentration increased. PDGF reduced bone formation at twenty-eight days in a dose-dependent manner. This inhibitory effect was resolved by fifty-six days, except in tissues in which demineralized bone matrix and 10 μg of PDGF had been implanted. In sites treated with 10 μg of PDGF, the area of new bone was decreased and the area of bone marrow was reduced at twenty-eight and fifty-six days. PDGF also appeared to retard resorption of demineralized bone matrix in a dose-dependent manner. Platelet-rich plasma reduced osteoinduction by human demineralized bone matrix that had high osteoinductive activity and had no effect on osteoinduction by demineralized bone matrix with low activity. Conclusions: PDGF inhibits, in a dose-dependent manner, intramuscular osteoinduction and chondrogenesis by demineralized bone matrix in immunocompromised mice. Platelet-rich plasma also reduces the osteoinductivity of active demineralized bone matrix. Clinical Relevance: Osteoinduction in the nude mouse may not reflect growth-factor effects in bone. However, these data indicate that PDGF and platelet-rich plasma should not be used with demineralized bone matrix if the purpose is to increase osteoinduction, although these substances may increase other aspects of bone-healing. Additional studies are needed to determine the clinical relevance of these observations.