In vitro biocompatibility of bioresorbable polymers: poly(L, DL-lactide) and poly(L-lactide-co-glycolide).

The acute toxicity of two degradable polymers, a 70:30 poly (L-D, L-lactide) (PLDLA) and a 90:10 poly(L-lactide-co-glycolide) (PLGA), was evaluated by the agar diffusion test and the filter test with L929 mouse fibroblasts. Extracts of the materials prepared in phosphate-buffered saline at 37 and 70 degrees C were assessed for mitochondrial succinate dehydrogenase activity by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT assay) and the incorporation of 5-bromo-2'-deoxyuridine (BrdU) into DNA of BALB 3T3 cells. Both materials revealed no signs of cytotoxicity during the agar diffusions and filter tests. In the MTT and BrdU assays PLDLA and PLGA showed similar results. Cells treated with extracts prepared at 37 degrees C caused slight stimulation of mitochondrial activity. In contrast, cells incubated with the 70 degrees C media revealed a concentration-dependent decrease of mitochondrial activity. DNA synthesis was significantly decreased by the 37 degrees C extracts. As in the MTT assay, the effect of the extracts prepared at 70 degrees C was significantly greater. From these in vitro results it is suggested that PLDLA and PLGA have satisfactory biocompatibility. High concentrations of the degradation products, however, had a toxic influence on the cell culture systems used.

New bioresorbable pin for the reduction of small bony fragments: design, mechanical properties and in vitro degradation.

The design, material properties, and in vivo degradation characteristics of a new resorbable pin for the reductions of small bony fragments are described. The Polypin, made of 70:30 poly (L, DL-lactide), had an initial bending strength of 155-163 MPa, as measured by a three-point bending test. Ethylene oxide (EO)- and gamma-sterilization did not substantially affect its initial mechanical properties. The initial molecular weight (Mw) of 523,000 to 600,000, however, decreased 60-75% after gamma-sterilization. Incubation of the EO-sterilized pins in 37 degrees C saline solution produced a complete loss of bending strength at 18 months. An accelerated test at 70 degrees C led to a complete loss of strength after only 96 h. Degradation of the gamma-sterilized pin at 70 degrees C was about 30% faster than that of the EO-sterilized pin. Bending strength and molecular weight were unaffected by storage at room temperature for 46 months. The relatively slow strength loss rate of the Polypin potentially extends the application of resorbable devices to slow-healing fractures. The new pin design allows application of light interfragmentary compression, thus reducing the risk of pin loosening, and an X-ray marker is provided.

IL-6 and PGE2 release by human osteoblasts on implant materials.

Regarding orthopaedic implant loosening it has been hypothesized that particle-activated macrophages release interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-alpha). This in turn stimulates osteoblasts to release interleukin-6 (IL-6) and prostaglandin E(2) (PGE(2)). These mediators recruit and activate osteoclasts and may therefore lead to bone resorption and loss of implant fixation. In this study we compared the ability of different materials to induce the release of IL-6 and PGE(2) from primary isolated, human osteoblasts without preceding activation by macrophages. We tested stainless steel, cobalt-chromium alloy (CoCrMo), commercially pure titanium (cpTi), Ti-6Al-7Nb and Ti-6Al-4V processed in the same manner as corresponding clinical implants. After 12 and 24h the cells had actively secreted IL-6 and PGE(2). There were no clear differences among the implant materials or with the plastic control. The amount of factors the cells released in our study compare well with the findings of other authors who investigated osteoblasts on plastic. In comparison with the literature these amounts are lower than secretion levels of osteoblasts stimulated with implant particles, IL-1 or TNF-alpha. Moreover, other authors found that osteoclasts require higher concentrations of PGE(2) to become activated than the concentrations measured in our experiments. Therefore, the amount of PGE(2) released from the osteoblasts in our study is probably not sufficient to induce osteolytic activity. Because of contradictory statements in the literature it is unclear if the measured IL-6 concentrations promote osteolytic activity. Differences in material composition does not significantly influence the release of these factors if the materials have similar surface roughnesses.

[Diclofenac inhibits proliferation and matrix formation of osteoblast cells]. / Diclofenac hemmt die Proliferation und Matrixbildung osteoblastärer Zellen.

Cell culture studies have shown that NSAID may influence osteogenic activities of osteoblast cultures. However, these studies did not consider long-term effects on differentiating cells. The influence of Voltaren with the non-steroidal agent diclofenac on proliferation and gene expression of the osteoblast-like cell line SaOS-2 was investigated 2, 9, and 16 days after incubation. Two days after 24 h of incubation, 50 microg/ml diclofenac reduced the proliferation and collagen type I expression while 9 and 16 days later no effect was found on either of the parameters. In contrast, 50 microg/ml NSAID has no effect on alkaline phosphatase expression 2 days after incubation while 9 and 16 days later expression had been reduced. Lower concentrations (1.56 and 0.19 microg/ml) had no effects on the studied parameters. BrdU and MTT test showed that 50 microg/ml diclofenac reduced proliferative and metabolic activity. Lower concentrations (< or =25 microg/ml) had a lower or no influence. The findings indicate that the NSAID impairment depends on cellular differentiation stage and is not confined to the time during or immediately after NSAID incubation. According to these results in vitro testing of drugs should be performed over a longer time period to detect possible long-term impacts.

Proliferation and differentiation parameters of human osteoblasts on titanium and steel surfaces.

Commercially pure titanium (cpTi), titanium alloys, and steel are often used for dental and orthopedic implants. In these applications titanium is considered the "gold standard." However, tissue reactions around titanium implants and the changing trend to leave orthopedic devices in the body have led to a new examination of the preferred material. This in vitro study tested the behavior of osteoblasts on cpTi, Ti-6Al-7Nb, and stainless steel with surface designs similar to clinical implants. After surface characterization by scanning electron microscopy and profilometry, cell proliferation and the differentiation parameters of alkaline phosphatase (ALP) activity and osteocalcin were measured. For all materials tested, the growth curves showed a similar kinetic. On Ti-6Al-7Nb, ALP activity was significantly lower when compared with steel, and cpTi and did not change over the time. ALP activity increased moderately on steel and cpTi. Osteocalcin levels were higher on both titanium materials than on steel. Based on undisturbed cell growth and the relatively high alkaline phosphatase and osteocalcin levels, we suggest that cpTi provides the best biocompatibility with regard to proliferation, in addition to more reliable early and late differentiation markers of human osteoblasts in vitro.

Degradation behavior of composite pins made of tricalcium phosphate and poly(L,DL-lactide).

Combining a bioactive ceramic with a resorbable polymer may improve the biocompatibility and the osseointegration of degradable fracture fixation devices. This study reports on the mechanical properties and degradation behavior of two composite pins made of poly(L,DL-lactide) and 10 and 30% beta-tricalcium phosphate (TCP), respectively. The pins were compared to a pin made of 100% poly(L,DL-lactide). The failure force, bending strength, and molecular weight were determined during in vitro degradation at 37 degrees C up to 78 weeks. The blending with 10 or 30% beta-TCP decreased the initial mechanical properties and led to an accelerated degradation rate. The pins with 30% TCP lost half of their strength after 16 weeks, i.e. faster than the unmodified pin (40 weeks). The pins with 10% TCP, however, showed a decreased initial strength (128+/-9 MPa) compared to the unmodified pin (152+/-9 MPa) but very similar degradation characteristics. The drop of the molecular weight was not significantly different between the three types of pins. It was concluded that the mechanical requirements for a pin for the fixation of small bony fragments with improved biocompatibility were fulfilled by the composite pin with 10% TCP but not by the pin with 30% TCP.

In vitro biocompatibility of resorbable experimental glass ceramics for bone substitutes.

Tricalcium phosphate ceramics (TCPs) are increasingly used as bone substitutes. They demonstrate good biocompatibility and degrade relatively slowly. New glass ceramics based on calcium alkali orthophosphates (Ca(2)KNa(PO(4))(2)) were developed that degrade faster than TCP but could have reduced biocompatibility due to their high solubility. Therefore, they were modified by a neutralizing surface treatment. The aim of this study was to evaluate the biocompatibility of some of these ceramics, GB1a, GB9, and GB14, which differ in the amount of added Na, K, Mg, or Si ions, with standard and modified surfaces. The in vitro cytotoxicity of the ceramics GB1a, GB9, and GB14 was determined by the agar diffusion and filter test and the microculture tetrazolium (MTT) assay. In order to investigate the influence of surface modification, these three ceramics were compared to their surface-treated counterparts, GB1aN, GB9N, and GB14N. GB1a, the ceramic with the highest in vitro solubility, showed the strongest toxic influence in all cell culture tests. GB9 and GB14 produced better results. In contrast, the counterparts with modified surfaces exhibited no (GB9N, GB14N) or weak (GB1aN) signs of cytotoxicity. It is concluded that the toxicity of the ceramics GB1a, GB9, and GB14 depends on their solubility. A positive influence of the surface treatment on in vitro biocompatibility was demonstrated. Therefore, the surface-treated glass ceramics could be promising materials for bone replacement.

A composite polymer/tricalcium phosphate membrane for guided bone regeneration in maxillofacial surgery.

The aim of the study was the development of a resorbable membrane for guided bone regeneration (GBR) with improved biocompatibility, which should be stiff enough to avoid membrane collapse during bone healing. Combining a bioactive ceramic with a resorbable polymer may improve the biocompatibility and osteoconductivity of resorbable devices. The present article describes the preparation, the mechanical properties, and the in vitro degradation characteristic of a composite membrane made of poly(L, DL-lactide) and alpha-tricalcium phosphate in comparison to a membrane made of pure poly(L, DL-lactide). The tensile strength and the elastic modulus as well as the molecular weight of the membranes were measured after in vitro degradation in buffer at 37 degrees C up to 28 weeks. The initial tensile strength of the composite and the polymer membrane was 37.3 +/- 2.4 MPa and 27.7 +/- 2.3 MPa and the elastic modulus 3106 +/- 108 MPa and 3101 +/- 104 MPa, respectively. The mechanical properties remained constant up to 8 weeks and then decreased slowly until week 28. The molecular weight of both membranes decreased steadily from 170,000 D to 30,000 D. It was concluded that the mechanical requirements for a membrane for GBR were fulfilled by the composite membrane.

Composites made of rapidly resorbable ceramics and poly(lactide) show adequate mechanical properties for use as bone substitute materials.

Porous composite materials made of poly(L, DL-lactide) and a ceramic component, alpha-tricalcium phosphate (alpha-TCP) or one of the rapidly resorbable glass ceramics, GB9N or GB14N, respectively, were developed to be used as bone substitutes. The present article describes the mechanical properties and the in vitro degradation characteristic of the different composite materials. The yield strength, the elastic modulus, and the molecular weight were measured after in vitro degradation up to 78 weeks. The initial strengths of the alpha-TCP composite (12.5 +/- 0.7 MPa) was higher than that of the GB9N and GB14N composites (8.3 +/- 0.2 MPa and 10.9 +/- 0.2 MPa, respectively). The initial elastic moduli of the composites were between 450 and 650 MPa. The mechanical properties remained constant until a degradation period of 26 weeks. Then they decreased continuously until they were completely lost at week 52. The molecular weight (M(w)) decreased steadily from 91,000 D in the case of the alpha-TCP composite and 78,000 D and 85,000 D in the case of the GB9N or GB14N composites, respectively, to about 10,000 D at week 78. It was concluded that the composites show adequate mechanical properties in the range of cancellous bone and a suitable degradation characteristic to be used as bone substitute materials.

In vivo investigations on composites made of resorbable ceramics and poly(lactide) used as bone graft substitutes.

Porous composites made of poly(L, DL-lactide) (PLA) and alpha-tricalcium phosphate (alpha-TCP) or the glass ceramic, GB14N, respectively, were investigated in a loaded implant model in sheep. Six, 12 and 24 months after implantation histological and biomechanical evaluation were performed and compared to autogenous bone transplants. No significant differences were observed between the composites. After 6 months, the interconnecting pores of the alpha-TCP-composite and the GB14N-composite were filled with newly formed bone (14 +/- 5% or 29 +/-15% of the implant, respectively) and soft tissue (30 +/-9% or 21 +/-12% of the implant, respectively). Only a mild inflammatory response was observed. The reaction was similar after 12 months. However, after 24 months a strong inflammatory reaction was seen. The newly formed bone was partly osteolytic. The adverse reaction occurred simultaneously to a significant reduction of the PLA component. The histological results were reflected by the biomechanical outcomes. Both composites showed compression strengths in the range of the autologous bone graft until 12 months of implantation. After 2 years, however, the strengths were significantly decreased. It is concluded that the new composites cannot yet be used for clinical application. An improvement in biocompatibility might be reached by a better coordination of the degradation times of the polymer and the ceramic component.

A new bioresorbable polymer for screw augmentation in the osteosynthesis of osteoporotic cancellous bone: a biomechanical evaluation.

The aim of the study was to assess the mechanical efficacy of a new resorbable polymer developed on the basis of alkylene bis(dilactoyl)-methacrylate to improve the anchorage of osteosynthesis material in cancellous bone. Cancellous bone screws were inserted in bovine as well as in human vertebrae and human femoral condyles and were augmented with the new polymer or polymethylmethacrylate (PMMA), respectively. Nonaugmented screws were used as controls. A removal torque test, a dynamic fatigue test, and a pullout test were performed. Augmentation with the new polymer increased the removal torque by 84% in human femoral bone. In the dynamic fatigue test of bovine vertebrae, the removal torque after cyclic loading was 115% higher for the new polymer compared to the nonaugmented controls. In the human vertebrae, the reinforcement with the new polymer increased the removal torque after dynamic loading by 114%. The augmentation with the new polymer increased the pullout force by 88% in bovine vertebrae and by 118% in human vertebrae in comparison to nonaugmented screws. It was concluded that augmentation by the new resorbable polymer significantly enhanced the anchorage of bone screws in cancellous bone. The mechanical efficiency of the new polymer was comparable to that of PMMA cement.