Lars Adolfsson

A Novel Biodegradable Delivery System for Bone Morphogenetic Protein-2

Background: The efficacy of recombinant growth factors in vivo is highly dependent on the delivery vehicle. The authors investigated the osteoinductive effects of recombinant human bone morphogenetic proteins (BMP)-2 implanted together with a complex of heparin and chitosan. Methods: Sixty rats were used. Three different carriers in gel formulation (type I collagen, heparin/type I collagen, and heparin/chitosan) were mixed with either 0, 10, or 50 &mgr;g of BMP-2, making the number of groups nine. The gels were injected into the quadriceps muscles of both legs in 45 rats (n = 10 per group). Freeze-dried formulations of the carriers were also tested with the same amounts of BMP-2 using 15 rats (n = 5 per group). Four weeks after implantation, the quality and amount of newly formed bone were assessed. Results: Chitosan was shown to protect the heparinase-mediated degradation of heparin in vitro. The osteoinductive effects of BMP-2 in combination with heparin/chitosan were superior as compared with BMP-2 implanted together with type I collagen. Interestingly, the heparin/chitosan complex induced a small amount of bone also without BMP-2 added. The heparin/chitosan was completely absorbed after 4 weeks as determined by histologic evaluation, and a normal active bone formation was present. The freeze-dried formulations of the carriers demonstrated similar osteoinductive effects as the gels. Conclusions: An osteoinductive formula for clinical use is needed for general bone reconstruction. Heparin in complex with chitosan has the ability to stabilize or activate the growth factor in vivo and induce the generation of new bone in good yields.

Cytokines in blood from septic patients interact with surface-immobilized heparin.

When passing blood from septic patients through a column packed with surface heparinized beads, we were able to significantly reduce concentrations of the proinflammatory cytokine tumor necrosis factor (TNF)-&agr; from initially very high levels. Passage of blood over nonheparinized beads did not affect the TNF levels. Meanwhile, concentrations of the regulated on activation, normal T-cells expressed, and secreted leukocyte activating cytokine (RANTES) remained unchanged following passage through the heparinized column, but rose significantly after passage through a column packed with the nonheparinized control beads. We conclude that surface heparinization may be a useful technique for selectively regulating the levels of heparin-binding cytokines from whole blood. This may have potential implications for the treatment of hyper-inflammatory conditions such as severe sepsis. Our data also suggests that surface activation and its associated inflammatory response may be avoided by using heparinization of the extracorporeal circuit.