Effect of polymer molecular weight on the bone biological activity of biodegradable polymer/calcium phosphate cement composites.

Previous studies demonstrated that the addition of biodegradable polymer microparticles to calcium phosphate (CaP) cement improves the cement's degradative behavior without affecting its handling characteristics, especially its injectability and moldability. We investigated the influence of molecular weight of polymeric microparticles included in CaP cement on implant degradation and bone formation in critical-sized defects. Forty rats received cranial defects filled with formulations of CaP cement and poly(DL-lactic-co-glycolic acid) (PLGA) microparticles. Microparticles consisted of 100% high- (HMW) or low-molecular-weight (LMW) PLGA or mixtures of these (25%, 50%, or 75%). Implantation time was 12 weeks. Porosity measurements showed that the 100% HMW group was significantly less porous than the other groups. Histology and histomorphometry revealed significantly greater implant degradation in the 100% LMW group. Defect bridging was mainly seen in the 75% and 100% LMW groups, with the highest amount of bone in the 100% LMW formulation. These results suggest that LMW PLGA microparticles are associated with better bone formation than HMW PLGA, which is most likely explained by the greater degradation of LMW PLGA microparticles. In conclusion, CaP cement composites with high percentages of LMW PLGA microparticles show good bone transductive behavior, with complete defect bridging. The 100% LMW group turned out to be the best formulation.

No increased bone formation around alendronate or omeprazole loaded bioactive bone cements in a femoral defect.

Alendronate and omeprazole have been found to influence bone healing by interfering with osteoclastic activity, resulting in increased bone formation. The biological effect of these conventional drugs, incorporated into bioactive bone cement (G2B1), was investigated in a rabbit model. The 2 materials and a control were inserted in defects created in the femoral condyle of rabbits. Implantation time was 6 and 12 weeks. After retrieval, micro-computed tomography and histomorphometry were performed to quantify bone mineral density (BMD) and bone volume (BV) of the implant-surrounding bone mass and the percentage of bone-to-implant contact. BMD and BV were similar in all groups. The percentage of bone-to-implant contact was significantly lower in the alendronate and omeprazole groups than in controls after 6 weeks of implantation. After 12 weeks, this difference in bone contact disappeared for the omeprazole but not for the alendronate implants, which were almost completely surrounded by a fibrous capsule, associated with a limited inflammatory response. In conclusion, in the current study, alendronate and omeprazole did not result in better bone healing when incorporated into bioactive bone cement than did plain control implants. Moreover, an additional cytotoxicity assay revealed that alendronate evoked a toxic response.

The kinetic and biological activity of different loaded rhBMP-2 calcium phosphate cement implants in rats.

The healing of large bone defects can be improved by osteogenic bone graft substitutes, due to growth factor inclusion. A sustained release of these growth factors provides more efficient bioactivity when compared with burst release and might reduce the dose required for bone regeneration, which is desirable for socioeconomical and safety reasons. In this study, we compared different rhBMP-2 loadings in a sustained release system of CaP cement and PLGA-microparticles and were able to couple kinetic to biological activity data. Fifty-two rats received a critical-size cranial defect, which was left open or filled with the cement composites. The implants consisted of plain, high, and five-fold lower dose rhBMP-2 groups. Implantation time was 4 and 12 weeks. Longitudinal in vivo release was monitored by scintigraphic imaging of (131)I-labeled rhBMP-2. Quantitative analysis of the scintigraphic images revealed a sustained release of (131)I-rhBMP-2 for both doses, with different release profiles between the two loadings. However, around 70% of the initial dose was retained in both implant formulations. Although low amounts of rhBMP-2 were released (2.4 +/- 0.8 mug in 5 weeks), histology showed defect bridging in the high-dose implants. Release out of the low-dose implants was not sufficient to enhance bone formation. Implant degradation was limited in all formulations, but was mainly seen in the high-dose group. Low amounts of sustained released rhBMP-2 were sufficient to bridge critically sized defects. A substantial amount of rhBMP-2 was retained in the implants because of the slow release rate and the limited degradation.

Closing capacity of segmental radius defects in rabbits.

In the research of synthetic bone graft substitutes, the relevance for bone regeneration can be confirmed in a critical-sized model. In this study the rabbit radial defect was investigated as an ingenious model of critical size, due to its defect immobilizing intact ulna. In addition, the influence of poly(DL-lactic-co-glycolic acid) (PLGA) on bone regeneration was determined. Sixteen, 4-month-old rabbits received bilateral segmental radial defects of 15 or 20 mm. The osteotomy ends were marked with small titanium pins. Half of the group received injected PLGA microparticle/carboxymethylcellulose implants. Implantation time was 12 weeks. Evaluation consisted of radiographs after surgery and sacrifice, microcomputed tomography and histology. The radiographs revealed that the created defects were significantly smaller after sacrifice. Further a number of radii showed fibrocartilaginous interposition. Both findings indicated instability of the created defect. All evaluation techniques revealed that 15 and 20 mm were not of critical size, as most defects were more or less regenerated. PLGA microparticles did not influence bone regeneration significantly. In conclusion, 15- and 20-mm radius defects in 4-month-old rabbits were not a suitable model for bone regeneration as these defects were neither critical size nor stable. PLGA-microparticle degradation did not influence bone regeneration.

Investigation as to the osteoinductivity of macroporous calcium phosphate cement in goats.

For bone formation in critical-sized or poor healing defects, osteoinductive behavior of synthetic bone grafts is crucial. Although the osteoconductive behavior of calcium phosphate (CaP) cement is generally accepted, its osteoinductive potential is less reported. In this study, osteoinduction of porous CaP cement was investigated. Four goats received each six subcutaneous placed prehardened porous CaP cement implants. Implantation time was 3 and 6 months. After explantation, histological evaluation and scoring with a histological grading scale for soft-tissue implants were performed. The histological sections revealed that the implants degraded for more than 50% over time. The implants had lost their macroporous structure from 3 months on. A medium-thick fibrous capsule with a few inflammatory cells surrounded the implants after 3 months. This capsule significantly decreased in thickness after 6 months. Throughout the implant ingrowth of fibrous tissue was seen with scattered foci of inflammatory cells. Cement particles were surrounded by a layer of inflammatory cells. The massive inflammatory response in the interstice was seen after 3 months, which disappeared after 6 months implantation. No bone formation was detected in any of the specimens. The fast degradation and thereby collapsing of the porous structure of our CaP cement implant might have prevented osteoinduction.

Bone regeneration of porous beta-tricalcium phosphate (Conduit TCP) and of biphasic calcium phosphate ceramic (Biosel) in trabecular defects in sheep.

In this study bone regeneration between porous beta-tricalcium phosphate (Conduit TCP) and biphasic calcium phosphate ceramic (Biosel), with a hydroxyapatite/beta-TCP ratio of 75/25, was compared. The ceramic particles were implanted in sheep trabecular bone for 3, 12, and 26 weeks. Histomorphometrical analysis revealed that Conduit degraded significantly during time and only 36% of the material was left at 26 weeks implantation time. Biosel, in contrast, remained nearly intact. The degradation of Conduit was due to dissolution as well as cell-mediated. Biosel showed a high cellular intervention, although this material did not degrade. Both materials were osteoconductive. The amount of newly formed bone appeared greater in the Conduit group after 26 weeks (46% +/- 8% as compared to 37% +/- 8% for Biosel), but this difference was not significant. Bone distribution over the defect was homogeneous in Conduit, whereas Biosel showed significantly more bone in the periphery of the defect after 26 weeks in comparison to the center. In conclusion, both ceramics are biocompatible and osteoconductive. Degradation showed a difference in amount and in cellular events, with more degraded Conduit TCP with less cellular intervention as compared to Biosel.

Osteochondral repair in the rabbit model utilizing bilayered, degradable oligo(poly(ethylene glycol) fumarate) hydrogel scaffolds.

In this study, hydrogel scaffolds, based on the polymer oligo(poly(ethylene glycol) fumarate) (OPF), were implanted into osteochondral defects in the rabbit model. Scaffolds consisted of two layers-a bottom, bone forming layer and a top, cartilage forming layer. Three scaffold formulations were implanted to assess how material composition and transforming growth factor-beta1 (TGF-beta1) loading affected osteochondral repair. Critical histological evaluation and scoring of the quantity and quality of tissue in the chondral and subchondral regions of defects was performed at 4 and 14 weeks. At both time points, no evidence of prolonged inflammation was observed, and healthy tissue was seen to infiltrate the defect area. The quality of this tissue improved over time with hyaline cartilage filling the chondral region and a mixture of trabecular and compact bone filling the subchondral region at 14 weeks. A promising degree of Safranin O staining and chondrocyte organization was observed in the newly formed surface tissue, while the underlying subchondral bone was completely integrated with the surrounding bone at 14 weeks. Material composition within the bottom, bone-forming layer did not appear to affect the rate of scaffold degradation or tissue filling. However, no bone upgrowth into the chondral region was observed with any scaffold formulation. TGF-beta1 loading in the top layer of scaffolds appeared to exert some therapeutic affect on tissue quality, but further studies are necessary for scaffold optimization. Yet, the excellent tissue filling and integration resulting from osteochondral implantation of these OPF-based scaffolds demonstrates their potential in cartilage repair strategies.

Donor-site morbidity after free vascularized autogenous fibular transfer: subjective and quantitative analyses.

The purpose of this study was to determine the subjective and quantitative donor-site morbidity after removal of a free vascularized fibula flap for autoreconstruction. Ten patients and six age-matched, healthy control subjects were included in this study. The postoperative periods ranged from 6 to 87 months. Subjective donor-site morbidity was assessed with a patient questionnaire and the Enneking system. For quantification of donor-site morbidity, gait was evaluated during normal walking, walking under visual and cognitive constraints, and walking at a velocity higher than the preferred one. In general, the patient perception of donor-site morbidity was low. Complaints were frequently mentioned, however, including pain (60 percent), dysesthesia (50 percent), a feeling of ankle instability (30 percent), and inability to run (20 percent). Gait analyses revealed that patients walked at a lower preferred velocity, compared with control subjects. Furthermore, they demonstrated significant increases in the coefficients of variation of stride time during walking under visual and cognitive loads and during walking at a velocity higher than the preferred one, compared with normal walking. These increases were not observed for control subjects. These findings suggest that the reautomatization of gait is affected among patients. This study demonstrates that fibula harvesting is associated with low subjective morbidity but frequent complaints. Walking during complex tasks and at high velocities reveals that restoration of gait is not complete after partial fibulectomy.