A novel, multi-barrier, drug eluting calcium sulfate/biphasic calcium phosphate biodegradable composite bone cement for treatment of experimental MRSA osteomyelitis in rabbit model.

This article discloses the development of an effective and versatile technology to prepare a novel antibiotics-loaded biodegradable composite bone cement to treat methicillin-resistant Staphylococcal (MRSA) osteomyelitis and reports its detail in vitro characterization, drug loading efficiency, physico-mechanical properties, drug elution in simulated body fluid (SBF) and human plasma, merits and demerits over poly-methyl methacrylate (PMMA) cement. Chronic osteomyelitis in rabbit tibia (42) was induced by MRSA and composite cement was implanted to evaluate its safety and efficacy over PMMA cement and parenteral treated animals with histopathology, radiographs, bone/plasma drugs concentration, and SEM for 90days. The composite cement showed higher setting time, degradability, pH rise, injectability, in vitro drug elution but lesser mechanical strength than PMMA cement. Antibiotics release from cement beads was faster in SBF than plasma. Further, in vivo antibiotics elution from composite (42days) showed effective concentration against MRSA without eliciting drug-toxicity. Platelets activation by composite was an extraordinary feature. The in vivo studies also proved the superiority of composite cement than other treatment methods in terms of faster infection control and osteosynthesis. Based particularly on drug elution and in vivo results, this newly developed cement can successfully be used in clinical cases of chronic osteomyelitis.

Influence of different modifications of a calcium phosphate bone cement on adhesion, proliferation, and osteogenic differentiation of human bone marrow stromal cells.

Collagen and noncollagenous proteins of the extracellular bone matrix are able to stimulate bone cell activities and bone healing. The modification of calcium phosphate bone cements used as temporary bone replacement materials with these proteins seems to be a promising approach to accelerate new bone formation. In this study, we investigated adhesion, proliferation, and osteogenic differentiation of human bone marrow stromal cells (hBMSC) on Biocement D/collagen composites which have been modified with osteocalcin and O-phospho-L-serine. Modification with osteocalcin was carried out by its addition to the cement precursor before setting as well as by functionalization of the cement samples after setting and sterilization. hBMSC were cultured on these samples for 28 days with and without osteogenic supplements. We found a positive impact especially of the phosphoserine-modifications but also of both osteocalcin-modifications on differentiation of hBMSC indicated by higher expression of the osteoblastic markers matrix metalloproteinase-13 and bone sialo protein II. For hBMSC cultured on phosphoserine-containing composites, an increased proliferation has been observed. However, in case of the osteocalcin-modified samples, only osteocalcin adsorbed after setting and sterilization of the cement samples was able to promote initial adhesion and proliferation of hBMSC. The addition of osteocalcin before setting results in a finer microstructure but the biological activity of osteocalcin might be impaired due to the sterilization process. Thus, our data indicate that the initial adhesion and proliferation of hBMSC is enhanced rather by the biological activity of osteocalcin than by the finer microstructure.

Altered bioreactivity and limited osteoconductivity of calcium sulfate-based bone cements in the osteoporotic rat spine.

BACKGROUND CONTEXT: Previous studies documenting the osteoconductive nature of calcium sulfate (CaSO(4))-based biomaterials have been largely limited to animal models exhibiting nonosteoporotic bone biology. In addition to diminished bone mineral density (BMD) and altered bone microarchitecture, the osteoporosis phenotype is associated with a proinflammatory and pro-osteolytic state. Thus, osteoporosis may elicit an amplified bioreactivity to common orthopedic biomaterials, potentially limiting their full osteoconductive capabilities in vivo. PURPOSE: The purpose of this study is to test the hypothesis that CaSO(4)-based bone cements exhibit altered bioreactivity and limited osteoconductivity in response to osteoporotic conditions. STUDY DESIGN: 1) Microcomputed tomography (micro-CT) radiomorphometry study and 2) histological analysis. METHODS: Our laboratory has previously established a preclinical model of osteoporosis using the rodent osteoporotic spine (OS). Caudal vertebral defects were filled with either CaSO(4) or CaSO(4)/CaPO(4) (Hybrid) cement for each group (n=4). Over 8 weeks, cement resorption profiles, BMD, average cortical thickness, average trabecular thickness, average trabecular spacing, and diaphyseal bone volume fraction were assessed via micro-CT radiomorphometry. Histological analysis was performed on vertebrae obtained postsurgery and at Week 8. RESULTS: Both materials displayed an accelerated cement resorption profile after implantation into the OS vertebrae. Hybrid cement exhibited slower resorption compared with that of CaSO(4) under both normal female rats (NL) and OS conditions. The cement-mediated bone augmentation observed in the NL spine was altered under OS conditions. CONCLUSIONS: This study suggests that cement bioreactivity is heightened and osteoconductivity may be limited in a preclinical model of the OS. The disparity between the two resorption profiles suggests that this accelerated cement resorption is a material-dependent phenomenon. The proinflammatory and pro-osteolytic bone environment associated with the osteoporosis disease state may contribute to the accelerated resorption and altered osteoconductivity exhibited by both materials. Future study of potential biomaterials intended for use within the OS may necessitate further exploration of the relationship between biomaterial performance and osteoporosis bone biology.

Repair of calvarial defects in rats by prefabricated hydroxyapatite cement implants.

The aim of the present study was to test the hypothesis that calvarial defects can be repaired by using preformed implants of calcium phosphate bone cement (CPBC) in rats. Sixty adult female Sprague-Dawley rats received full-thickness calvarial nonhealing defects with a diameter of 8 mm. Three different CPBCs were used: group 1: tetracalcium phosphate-based powder; group 2: a blend of amorphous and crystalline calcium phosphate precursors; and group 3: an alpha-tricalcium phosphate (alpha-TCP)-based powder. Implants were left to cure for 25-40 min at room temperature in a silicon mold of 7.9 mm and inserted press fit into the defects. Fifteen animals served as unfilled controls. After 13, 26, and 52 weeks, the material was analyzed qualitatively by using surface-stained undecalcified thick-section specimens and quantitatively by using semiautomated histometry. Kruskal-Wallis tests were applied to compare mean values of periimplant bone formation at a significance level of p < 0.05. Three implants of group 1 fractured during insertion. Resorption of CPBC without complementary bone formation was noticed in these implants. Unfractured implants were resorbed with simultaneous apposition of bone on the implant surface. After 52 weeks, the resorption rate varied between 23.1 and 39.3%. Periimplant bone formation increased continuously on average around all implant types, but it reached statistical significance only in group 2. The results showed that repair of calvarial defects can be achieved by preformed CPBC implants. The rate of resorption of preformed implants is, however, much lower than that reported for in vivo cured CPBC.

Biological evaluation of an ionomeric bone cement by osteoblast cell culture methods.

Periosteal derived bovine osteoblast-like cells migrated in culture onto an ionomeric cement. Cell cultures were maintained for 4 weeks and used to study the in vitro behaviour of cells on the ionomeric bone cement (IC). The cells produced bone matrix proteins (osteocalcin, bone sialoprotein II) and were osteoblast-like. The osteoblast-like cells colonized the substrate in monolayers and produced an extracellular matrix as seen by light and scanning electron microscopy. Morphological comparison between cells growing on the ionomeric bone cement and cortical bone revealed no significant difference in phenotypic expression. Staining for aluminium in osteoblasts growing on the IC showed an uptake and storage of aluminium in the cells. Energy dispersive X-ray microanalysis revealed high concentrations of aluminium and silicon in the periosteal tissue. Despite the known toxic effect of aluminium in vivo and in vitro on osteoblasts, no signs of toxicity were apparent on light and scanning electron microscopy analysis.

Biomechanical evaluation of a biodegradable composite as an adjunct to internal fixation of proximal femur fractures.

Internal fixation of comminuted unstable fractures of the severely osteoporotic proximal femur is sometimes supplemented with polymethyl-methacrylate (PMMA). We here report an in vitro biomechanical evaluation of a biodegradable particulate composite that might be used for similar purposes. The composite includes a matrix phase consisting of a hydrolyzable prepolymer [polypropylene fumarate (PPF)] cross-linked with methacrylate monomer, and a particulate phase consisting of tricalcium phosphate and calcium carbonate. We implanted dynamic hip screws in 22 cadaveric proximal femora and measured the yield load for an oblique force applied to the femoral head. The hip screws were then reinforced with either PMMA or the PPF composite and tested again. On the basis of analysis of variance, the average increases in yield load for PMMA and PPF reinforcement of 1,750 and 1,130 N were statistically significant (p less than 0.00005), suggesting that both materials enhance congruence between implant and bone and thereby increase the projected load-bearing area of the implant. The increase in yield force with PMMA was slightly higher than the increase with PPF (p less than 0.05), but both values after reinforcement were close (3,790 +/- 561 N for PMMA vs. 3,240 +/- 669 N for PPF). If we can demonstrate that appropriate rates of degradation, bony ingrowth, and static and fatigue properties can be achieved in vivo with this system, our data suggest that this PPF composite may have potential as an adjunct to the internal fixation of unstable fractures of the osteoporotic hip.