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Study Design: An experimental study of cardiovascular complications arising during vertebroplasty (VP) of multiple levels in sheep., Objectives: To investigate the effect of pressurizing vertebral bodies during VP using different materials in the development of fat embolism (FE) and any associated cardiovascular changes., Summary of Background Data: Polymethylmethacrylate (PMMA) is the material of choice for VP. However, PMMA has several disadvantages, such as toxicity, exothermic curing, uncertain long-term biomechanical effects, and biocompatibility. Alternative materials are being developed for VP; however, there is the concern that an increase in intraosseous pressure and displacement of bone marrow contents could lead to FE and hypotension during VP regardless of what type of materials is used., Methods: In 20 sheep, four vertebral bodies were augmented either with PMMA or bone wax. Heart rate; arterial, central venous, and pulmonary artery pressure; cardiac output; and blood gas values were recorded. Postmortem the lungs were subjected to histologic evaluation., Results: The consecutive augmentation of four vertebral bodies with PMMA induced a cumulative FE that gradually deteriorated baseline mean arterial blood pressure (MABP) and blood gas values. The augmentation with bone wax resulted in similar cardiovascular changes and amount of intravascular fat in the lungs., Conclusion: There are potential cardiovascular complications during VP of multiple levels regardless of the augmentation material used. The deteriorating baseline MABP during VP is associated with the pressurization of the vertebral bodies rather than with the use of polymethylmethacrylate.

Study Design: A biomechanical cadaveric study of thoracic and lumbar vertebrae with simulated metastases quantifying intravertebral pressures during transpedicular vertebroplasty., Objective: To compare intravertebral pressures during percutaneous vertebroplasty in vertebrae with and without simulated lytic metastases., Summary of Background Data: Percutaneous vertebroplasty is designed to provide stability to vertebrae weakened by osteoporosis or metastatic disease. The complication rate is higher when the procedure is used for the treatment of lytic vertebral lesions. The major complications reported are radiculopathy, spinal cord compression, and embolic phenomena., Methods: Ten fresh-frozen cadaveric vertebrae were tested intact (7 lumbar, 3 thoracic) and 7 were tested with simulated lytic defects (4 lumbar, 3 thoracic). Defects were created by replacing a core of cancellous bone with soft tumor tissue in the center of the vertebral body. Simplex P (Howmedica Osteonics, Mahwah, NJ) cement was injected into each vertebra through a unipedicular approach at a constant rate of 3 mL per minute. Cement volume, injection force, and intravertebral pressures at the posterior vertebral body wall were recorded. Following the procedure, the vertebrae were sectioned to visualize cement and tumor disbursement., Results: There was no significant difference between the two groups for age, size, trabecular density, and cement volume. Vertebrae with simulated metastases generated an average maximum pressure of 39.66 kPa during cement injection versus 6.83 kPa in intact vertebrae (P < 0.05). Higher pressures were also generated in smaller vertebrae based on a power relationship (r2 = 0.71 intact, r2 = 0.43 tumor)., Conclusions: Percutaneous vertebroplasty produces higher intravertebral pressures in vertebrae containing a simulated lytic metastasis than in intact vertebrae. Pressures generated in the tumor specimens are sufficiently elevated to cause embolic phenomena.

Aim: In an experimental study the hydroxyapatite cement BoneSource was tested for the ability in relation to the defect size and for its resorption properties., Material and Methods: In an animal study, BoneSource was applied to repair bicortical defects of different sizes in frontal bones of six Goettingen minipigs. The area was evaluated radiographically and histologically 12, 18 and 40 weeks postoperatively., Results: After 40 weeks approximately 90% of the hydroxyapatite cement had been resorbed and replaced with bone. After 12 weeks, approximately 30% had been degraded, and 40% after 18 weeks. In small bone defects, the mucosa of the frontal sinus lined the bone substitute (BoneSource). In the reconstruction of large areas, a membrane was used to stabilize the material. Despite this membranous support, BoneSource material prolapsed into the frontal sinus., Discussion: Hydroxyapatite cement is for the repair of bone defects. It can be moulded to shape the reconstruction. Its use is limited by the defect size and the need for a dry bed., Conclusion: BoneSource is well suited to repair small defects with proven high biocompatibility. However, in large defects, the material is not sufficiently stable.

Study Design: The effect of cement augmentation on an osteoporotic lumbar functional spinal unit was investigated using finite-element analysis., Objective: To evaluate the influence of cement augmentation on load transfer, stresses, and strains., Summary of Background Data: Osteoporosis is the most frequent skeletal disease of the elderly, leading to weakness of the bony structures. Cement injection into vertebral bodies has been used to treat osteoporotic compression fractures of the spine. The clinical results are encouraging. Experimental biomechanical studies showed significant increases in stiffness and strength of treated bodies. However, little is known about the consequences for the adjacent, nontreated levels., Methods: Three-dimensional finite-element models of L2-L3 were developed and the material properties adapted to simulate osteoporosis. The influence of augmentation level as well as uni- and bipedicular filling with polymethylmethacrylate were investigated. Compression, flexion, and lateral bending were simulated., Results: Augmentation increased the pressure in the nucleus pulposus and the deflection of the adjacent endplate. The stresses and strains in the vertebrae next to an augmentation were increased, and their distribution was changed. Larger areas were subjected to higher stresses and strains. The treatment clearly altered the load transfer. Changes to the overall stress and strain distribution were less pronounced for unipedicular augmentation., Conclusions: Cement augmentation restores the strength of treated vertebrae, but leads to increased endplate bulge and an altered load transfer in adjacent vertebrae. This supports the hypothesis that rigid cement augmentation may facilitate the subsequent collapse of adjacent vertebrae. Further study is required to determine the optimal reinforcement material and filling volume to minimize this effect.

Calcium phosphate (CAP) rods and pastes based on dicalcium and tetracalcium phosphate chemistry were modified with a specific osteoclast proton pump inhibitor, Bafilomycin A(1), and implanted in the distal femurs of young male Wistar rats for 7, 10, and 14 days. The extent of osteoclastic resorption of these materials and the amount of regenerative bone formed were qualitatively evaluated. Resorptive activity similar to the remodeling process of natural bone was observed in the controls. In contrast, the resorption of materials containing Bafilomycin A(1) was considerably lower, even though tartrate-resistant acid phosphatase-positive osteoclasts were present at the tissue/material interface. A greater amount of newly formed bone consistently surrounded the CAP rods and pastes containing Bafilomycin A(1) in comparison to the control specimens, which indicated that this reparative bone was not resorbed as quickly as the new bone surrounding the controls. Increased local bone mass around the Bafilomycin A(1)-modified materials resulted from the diffusion of the proton pump inhibitor into the biological tissue at the defect site. Thus, through inhibition of the osteoclast proton pump, Bafilomycin A(1) slowed down not only the resorption of the implant material but also the resorption of the newly formed reparative bone, which resulted in an increased local bone mass.

The aim of this study was to evaluate biologic behavior of a composite of bioactive glass (BAG) (S53P4) and copolymer of poly(epsilon-caprolactone-co-DL-lactide) in experimental bone defects in rabbits. Twenty New Zealand white rabbits were used for the study. Bone defects (4 x 6mm) were prepared in the medial surfaces of the femoral condyles and the tibia. Cavities were filled with three different composites: composite with 60 wt% of small BAG granules (granule size <45 microm) and composites with 40 and 60 wt% of large BAG granules (granule size 90-315 microm). Copolymer without BAG was used as a reference material. Follow-up period was 8 and 16 weeks. In the femur at 8 weeks all the samples were partly surrounded by fibrous capsule. New bone formation was noticed in the areas where glass granules were in direct contact with the bone. At 16 weeks fibrous capsule was thinner in all samples. Bone ingrowth was found in the superficial layers of the composites with large glass granules. However, the percent of direct bone contact decreased between 8 and 16 weeks (p < 0.05). In the tibia at 8 weeks all the samples showed fibrous encapsulation. At 16 weeks fibrous capsules were thinner or occasionally disappeared. Bone ingrowth was noticed in the samples with large glass granules. Further, new bone formation was found in the medullary cavity. No signs of polymer degradation were seen at any time point. It can be concluded that the composite of BAG (S53P4) and copolymer of poly(epsilon-caprolactone-co-DL-lactide) is biocompatible with the bone tissue within the 16 weeks implantation period., (Copyright 2003 Elsevier Science Ltd.)

We conducted an in vivo experiment to evaluate the resorption rate of a calcium phosphate cement (CPC) with macropores larger than 100 microm, using the CPC called Biocement D (Merck Biomaterial, Darmstadt, Germany), which after setting only shows pores smaller than 1 microm. The gas bubble method used during the setting process created macroporosity. Preset nonporous and porous cement implants were inserted into the trabecular bone of the tibial metaphysis of goats. The size of the preset implants was 6 mm and the diameter of the drill hole was 6.3 mm, leaving a gap of 0.3 mm between implant surface and drill wall. After 2 and 10 weeks, the animals were euthanized and cement implants with surrounding bone were retrieved for histologic evaluation. Light microscopy at 2 weeks revealed that the nonporous implants were surrounded by connective tissue. On the cement surface, we observed a monolayer of multinucleated cells. Ten weeks after implantation, the nonporous implants were still surrounded by connective tissue. However, a thin layer of bone now covered the implant surface. No sign of cement resorption was observed. In contrast, the porous cement evoked a completely different bone response. At 2 weeks, bone formation had already occurred inside the implant porosity. Bone formation even appeared to occur as a result of osteoinduction. Also, at their outer surface, the porous implants were completely surrounded by bone. At 2 weeks, about 31% of the initial cement was resorbed. After 10 weeks, 81% of the initial phosphate cement was resorbed and new bone was deposited. On the basis of these observations, we conclude that the creation of macropores can significantly improve the resorption rate of CPC. This increased degradation is associated with almost complete bone replacement., (Copyright 2003 Wiley Periodicals, Inc.)

The aim of this study was to investigate the physicochemical and biological properties of a newly developed calcium phosphate cement (CaP cement) implanted in cortical bone. CaP cement was injected as a paste into tibia cortical bone defects in goats. Polymethylmethacrylate (PMMA) bone cement was used as a control. The animals were killed after 3 days, 2, 8, 16 and 24 weeks. X-ray diffraction and Fourier transform infrared spectroscopy performed at retrieved samples showed that the CaP cement had set as a carbonate apatite and remained stable over time. Light microscopic evaluation showed that after 2 weeks the cement was in tight contact with the bone without any inflammatory reaction or fibrous encapsulation. At later time points, the CaP cement implants were totally covered by a thin layer of bone and osteoclasts, present at the interface, which were clearly resorbing the cement. At locations where CaP cement was resorbed, new bone was deposited. Transmission electron microscopy revealed that indeed a seamless contact existed between CaP cement and bone, as characterized by the occurrence of an electron dense line of 50-60 nm thick that covered the CaP cement. Osteoblasts, in contact with the cement, were depositing new bone. Although the bulk of the material was still in situ after 24 weeks, the progressive osteoclast resorption of the cement followed by new bone formation suggests that all of the material may be replaced eventually. In contrast to the CaP cement, the PMMA reference cement was always surrounded by a thin fibrous capsule. The results indicate that the investigated CaP cement is biocompatible, osteoconductive as well as osteotransductive and is a candidate material for use as a bone substitute.

In this study, the soft tissue reaction to two newly developed injectable calcium phosphate bone cements (cement D and W) was evaluated after implantation in the back of goats. For one of the cements (cement D) the tissue reaction was also investigated after varying the concentration of accelerator Na(2)HPO(4) in the cement liquid (resulting in cement D1 and D2). Eight healthy mature female Saanen goats were used. The cement was applied 10min after mixing while it was still moldable and plastic. The material was given a standardized cylindrical shape. Thirty-two implants of each cement formulation were inserted and left in place for 1, 2, 4, and 8weeks. At the end of the study, eight specimens of each material and healing period were available for further analysis. Two specimens were used for X-ray diffraction (XRD) and Fourier Transform Infrared Spectroscopy (FTIR) and six specimens were used for light microscopical evaluation. XRD and FTIR showed that the cements did set as microcrystalline carbonate apatite with the disappearance of monetite from the cements during implantation. Histological analysis showed that after 8weeks of implantation around all materials a thin soft-tissue capsule was formed (thickness ranging from 5 to 15 cell layers) with almost complete absence of inflammatory cells. Only in some specimens a slightly higher inflammatory reaction was observed. This was due to cement surface defects and a zone of dispersed particles near the cement-soft tissue interface. There was almost no resorption of the material after 8 weeks of implantation. In a few 4 and 8weeks samples, small areas of calcification were found in the fibrous capsule surrounding the implants. On the basis of our observations, we conclude that the tested cements were biocompatible and can be used next to soft tissue.

Calcium phosphate cements (CPCs) augment the healing of bone following craniofacial and maxillofacial surgery. Their ability to function as bone substitutes has accelerated the healing processes from bone fractures or defects. Although CPCs all possess the same general function, small chemical composition differences in these compounds have a profound impact on their in vivo character. The main purpose of this article is to report the differences of several major CPCs. When available, specific clinical trials are referenced. The goal is to expand the knowledge base on CPCs to better inform the clinician and researcher.

The osteoinductivity of demineralized bone matrix (DBM) becomes significantly reduced if the specimens are further delipidated with chloroform-methanol. The addition of phosphatidylcholine (PC), a major constituent of the lipid fraction present in the calcification front during normal bone formation, can restore the biological activity. Active endochondral bone formation is observed in the DBM/PC implants placed in the anterior abdominal wall musculature or subcutaneously for 28 days. When PC is added to generate a putty containing 60% PC and 40% DBM, biochemical parameters associated with osteoinductivity are significantly enhanced. Biological responses evaluated histologically and by determination of alkaline phosphatase activity are in very good agreement. The DBM/PC putty has good handling properties, can be molded into different shapes, and does not wash away from the application site. An advantage of adding PC is that it not only enhances the handling properties, but also boosts the osteoinductivity of the preparation.

For more than 50 years PMMA bone cements have been used in orthopaedic surgery. In this study attempts were made to show whether cultured human bone marrow cells (HBMC) show an osteogenetic response resulting in new bone formation, production of extracellular matrix (ECM) and cell differentiation when they were cultured onto polymerized polymethylmethacrylate (PMMA)-hydroxyapatite (HA), conventional PMMA bone cement being taken as reference. Biocompatibility parameters were collagen-I and -II synthesis, the detection of the osteoblast markers alkaline phosphatase (ALP) and osteocalcin, the number of adherent cells and the cytodifferentiation of immunocompetent cells. Cement surface structure, HA stability in culture medium and chemical element analysis of specimens were considered. Fresh marrow cells were obtained from the human femora during hip replacement. Incubation time was up to ten weeks. We used atomic forced microscopy (AFM) and scanning electron microscopy (SEM) for cement specimen analysis. Fluorescent activated cell sorter (FACS), immunohistochemical staining. SEM and light microscopy (LM) served us to judge the cellular morphology. Products of the extracellular matrix were analyzed by protein dot blot analysis, SEM energy dispersive X-ray analysis (SEM-EDX) and Ca2+/PO(4)3- detection. HA particles increased the osteogenetic potential of PMMA bone cement regarding the cellular production of collagen, alkaline phosphatase (AP), the number of osteoblasts and the cellular differentiation pattern in vitro. Both tested cements showed good biocompatibility in a human long-term bone marrow cell-culture system.

The BiCONTACT endoprosthesis system, in addition to having a distinctly improved material structure (hardness, lubricant couple, reduction of abrasions, etc.), demonstrates characteristics geared towards providing uniform, optimal treatment under economically favourable conditions for the widest possible stock of patients. Furthermore, it secures local stability and freedom from pain as early as possible, sparing or--even better--building bone with a biologically orientated implantation technique and obtaining correspondingly acceptable medium- and long-term results via an appropriate surface coating for osseo-integration. The concept of the hip endoprosthesis, its design and implantation technique--which have not required modifications since its introduction in clinical practice 15 years ago--is capable of doing justice to all the individual, biological, instrumental and economic particularities mentioned.

The effects of phosphorylated chitin (P-chitin) on the tissue responses to two kinds of calcium phosphate cements (CPCs) were investigated using experimental rabbits. One of them consisting of monocalcium phosphate monohydrate, calcium oxide, 1 M phosphate buffer (pH: 7.4) and different amounts of P-chitin (CPC-I or P-CPC-I) with relatively neutral initial pH was filled as paste into tibial defects of the rabbits for 1, 4, 12 and 22 weeks. The other kind of cement made from dicacium phosphate dihydrate/calcium hydroxide/1 M Na2HPO4/different amounts of P-chitin (CPC-II or P-CPC-II) with relatively higher initial pH was implanted as prehardened cylinders into the radial defects of the rabbits for the same periods. Pure CPC-I and CPC-II were used as controls. Histological and histomorphological studies were performed on thin un-decalcified and decalcified sections. Three different bone formation types in the resorption lacuna of the P-CPCs were found during this study. The biodegradation rate of the P-CPCs had a negative relationship with the P-chitin content. Most of the low P-chitin-containing samples were bioabsorbed in 16 weeks, while the high P-chitin-containing samples disappeared in 22 weeks. The newly formed bone was identified with back scattered scanning electron microscopy and X-ray energy-dispersive spectrometry. The results show that with P-chitin component in a certain range, the P-CPCs are biocompatible, bioabsorbable and osteoinductive and could be used as promising candidates of bone repair materials.

Study Design: Transpedicular lumbar interbody fusion (TLIF) was performed in a sheep model comparing three treatment groups: a composite of osteogenic protein (OP)-1 and hydroxyapatite carrier (HA), HA without OP-1, and autograft., Objective: To evaluate the efficacy of the composite of OP-1 and HA (HA-OP-1) in achieving reliable TLIF., Summary of Background Data: Anterior fusion techniques directly address disc-related problems and achieve primary axial stability. However, they are characterized by high morbidity. Alternatively, the theoretically advantageous posterior TLIF technique using autograft fails clinically because it lacks compressive stability., Methods: In 36 sheep, lumbar vertebrae L4 to L6 were instrumented posteriorly. Endoscopically assisted TLIF of L4 to L5 was performed. In 12 sheep, the defect was filled with injectable HA-OP-1. Another 12 sheep were treated with HA and another 12 with autograft. Animals were killed at 8 weeks and evaluated by radiologic, histologic, and histomorphometric analysis and by fluorochrome labeling., Results: Only 10 autograft sheep were available for evaluation. Radiologically and histologically, TLIF with HA-OP-1 led to a fusion rate of 10 in 12 compared with autograft (one in 10 fused) and HA (two in 12 fused) ( = 0.0016). Semiquantitative radiologic and histologic scoring also revealed significant differences with superiority of HA-OP-1 ( = 0.0011). Compared with HA, HA-OP-1 presented significantly more ossification at the bone-cement interface ( = 0.0003) and less cement resorption ( = 0.0209). In four of 12 HA sheep, excessive resorption was responsible for local aseptic inflammation., Conclusions: Biointegration of the osteoconductive HA does not occur, because shear forces cause early HA fracture, subsequent fragmentation, and gross resorption (initiating severe inflammation in four of 12 sheep). In contrast, osteoinductive effects of HA-OP-1 enable bio-integration, resulting in full osseous composite sheathing and solid fusion. By use of this composite, TLIF is successfully applied in sheep. Harvesting autograft and the anterior approach are avoided.

Percutaneous vertebroplasty (PVP) is a radiologically guided therapeutic procedure, which consists of percutaneous injection of a liquid polymer (bone cement) into a destroyed vertebral body. PVP was invented in 1984, in France, first for treating vertebral body haemangioma. Since its introduction the indications have been expanded progressively and today PVP is indicated mainly for treatment of vertebral haemangioma, malignant vertebral tumor and osteoporotic vertebral compression fracture. The unique advantage of this technique is that besides the stabilization of the vertebral body--and partly in connection with this--it affords prompt and lasting pain relief. Based on published data the success rate of the procedure is 80-100% with a complication rate of 1-10%. Thus, PVP is a valuable minimally invasive tool, providing immediate pain relief and early mobility in carefully selected patients. However, further work is needed to define the benefits of PVP compared to the standard treatment. The purpose of this paper is to demonstrate the technique by analyzing scientific reports published to date and summarizing the first author's own experience gained at the University Hospital of Geneva, Department of Neuroradiology, Switzerland.

The development of tissue engineered materials for the treatment of large bone defects would provide attractive alternatives to the autografts, allografts, non-degradable polymers, ceramics, and metals that are currently used in clinical settings. To this end, poly(propylene fumarate) (PPF), a viscous polyester synthesized from diethyl fumarate (DEF), has been studied for use as an engineered bone graft. We have investigated the photocrosslinking of PPF dissolved in its precursor, DEF, using the photoinitiator bis(2,4,6-trimethylbenzoyl) phenylphosphine oxide (BAPO) and low levels of ultraviolet light exposure. A three factor, 2 x 2 x 4 factorial design was developed, studying the effects of PPF number average molecular weight, BAPO initiator content, and DEF content upon photocrosslinking characteristics and mechanical properties. Uncured DEF/PPF solution viscosity fell over three orders of magnitude as DEF content was increased from 0% to 75%. The exothermic photocrosslinking reaction released low levels of heat, with no more than 160J/g released from any formulation tested. As a result, the maximum photocrosslinking temperature remained below 47 degrees C for all samples. Both sol fraction and swelling degree generally increased with increasing DEF content. Compressive mechanical properties were within the range of trabecular bone, with the strongest samples possessing an elastic modulus of 195.3 +/- 17.5 MPa and a fracture strength of 68.8 +/- 9.4MPa. Finally, the results indicate that PPF crosslinking was facilitated at low DEF precursor concentrations, but hindered at higher precursor concentrations. These novel DEF/PPF solutions may be preferred over pure PPF as the basis for an engineered bone graft because they (1) exhibit reduced viscosity and thus are easily handled, (2) form polymer networks with compressive strength at fracture suitable for consideration for trabecular bone replacement, and (3) may be readily fabricated into solids with a wide range of structures.

Hydroxyapatite cement (HAC) has been shown to osseoconduct and may even osseoconvert. These properties render it suitable for repairing cranial defects in growing children. The objective of this pilot study was to determine if growth is restricted after HAC reconstruction in a growing, nonprimate, animal model. Frontoparietal craniotomies were performed on 10 4-week-old Yorkshire piglets. In 3 control piglets, the bone flap was resutured; in 7 experimental piglets, the defect was reconstructed with HAC. Analysis 6 months after surgery included craniometric measurements, biomechanical testing, and histological sectioning. Nine craniofacial measurements did not differ between controls and experimental animals. The mean midline-to-temporal line distance contralateral to the defect was 20% larger in the experimental group (P = 0.006), however, and experimental animals had a larger difference between right and left orbital breadths (+3% difference versus -1% difference; P = 0.003). The mean stiffness of the HAC-repaired defect was not different from the contralateral side in contrast to the resutured bone flap in controls, which was significantly less stiff than unoperated bone (162 N/mm compared with 358 N/mm; P < 0.05). Based on our animal model, HAC seems to be a sound alternative method of craniotomy reconstruction in the growing skull.

Acrylic bone cement formulations with antioxidant character were prepared by incorporation of a methacrylic monomer derived from vitamin E (MVE). Increasing concentrations of this monomer provided decreasing peak temperature values, ranging from 62 to 36 degrees C, and increasing setting time with values between 17 and 25 min. Mechanical properties were evaluated by compression and tension tests. Compressive strength of the new formulations were superior to 70 MPa in all cases. The cement containing 25 wt % MVE, however, showed a significant decrease in tensile properties. Biocompatibility of the new formulations was studied in vitro. The analysis of the effect of leachables from cements into the media showed continued cell proliferation and cell viability with a significant increase for the cement containing 15 wt % MVE. This formulation also showed a significant increase in cellular proliferation over a period of 7 days as indicated by the Alamar Blue test. The cells were able to differentiate and express phenotypical markers in presence of all materials. A significant increase in alkaline phosphatase activity was observed on the cements prepared in presence of 15-25 wt % MVE compared with PMMA. Morphological assessment showed that the human osteoblast (HOB) cells were able to adhere, retain their morphology, and proliferate on all the cements., (Copyright 2002 Wiley Periodicals, Inc.)

Study Design: Destructive biomechanical tests using fresh cadaveric thoracolumbar vertebral bodies., Objectives: To evaluate the compression strength of human vertebral bodies injected with a new calcium phosphate (CaP) cement with improved infiltration properties for augmentation of the vertebral bodies before compression fracture and also for vertebroplasty in comparison with polymethylmethacrylate (PMMA) injection., Summary of Background Data: Vertebroplasty is the percutaneous injection of PMMA cement into the vertebral body. While PMMA has high mechanical strength, it cures fast and thus allows only a short handling time. Other potential problems of using PMMA injection may include damage to surrounding tissues by a high polymerization temperature or by the unreacted toxic monomer, and the lack of long-term biocompatibility. Bone mineral cements, such as calcium carbonate and CaP cements, have longer working time and low thermal effect. They are also biodegradable while having a good mechanical strength. However, the viscosity of injectable mineral cements is high, and the infiltration of these cements into vertebral body has been questioned. Recently, the infiltration properties of a CaP cement have been significantly improved, which is ideal for the transpedicular injection to the vertebral bodies for vertebroplasty or augmentation of osteoporotic vertebral body strength., Methods: The bone mineral densities of 30 vertebral bodies (T2-L1) were measured using dual-energy x-ray absorptiometry. Ten control specimens were compressed at a loading rate of 15 mm/min to 50% of their original height. The other specimens had 6 mL of PMMA (n = 10) or the new CaP (n = 10) cement injected through the bilateral pedicle approach before being loaded in compression. Additionally, after the control specimens had been compressed, they were injected with either CaP (n = 5) or PMMA (n = 5) cement using the same technique, to simulate vertebroplasty. Loading experiments were repeated with the displacement control of 50% vertebral height. Load to failure was compared among groups and analyzed using analysis of variance., Results: Mean bone mineral densities of all five groups were similar and ranged from 0.56 to 0.89 g/cm2. The size of the vertebral body and the amount of cement injected were similar in all groups. Load to failure values for PMMA, the new CaP, and vertebroplasty PMMA were significantly greater than that of control. Load to failure of the vertebroplasty CaP group was higher than control but not statistically significant. The mean stiffness of the vertebroplasty CaP group was significantly smaller than control, PMMA, and the new CaP groups. The mean height gains after injection of the new CaP and PMMA cements for vertebroplasty were minimal (3.56% and 2.01%, respectively)., Conclusion: Results of this study demonstrated that the new CaP cement can be injected and infiltrates easily into the vertebral body. It was also found that injection of the new CaP cement can improve the strength of a fractured vertebral body to at least the level of its intact strength. Thus, the new CaP cement may be a good alternative to PMMA cement for vertebroplasty, although further in vivo animal and clinical studies should be done. Furthermore, the new CaP may be more effective in augmenting the strength of osteoporotic vertebral bodies for preventing compression fractures considering our biomechanical testing data and the known potential for biodegradability of the new CaP cement.

Background Context: The use of rigid instrumentation combined with bone graft makes intuitive sense given the requirements for vascular ingrowth, bone formation and a stable environment for the cellular events of healing to develop. However, with the advances of potent osteoinductive growth factors, the role of internal fixation may come into question. Whether bone morphogenic proteins (BMPs) would benefit from a more "stable" spinal segment for bone production and modeling remains unknown. In addition, it is unknown whether BMP and rigid fixation may have an additive effect on fusion healing., Purpose: This study is proposed to test the hypothesis that rigid fixation in the lumbar spine would be advantageous to achieve fusion for autogenous bone grafting, but fusion would occur regardless of fixation with the use of osteogenic protein (OP)-1., Study Design/setting: A histologic and radiographic analysis of BMP in a rabbit lumbar fusion model., Methods: Thirty-two rabbits were randomized into four groups: 1) control animals: in situ posterolateral L5-L6 arthrodesis using autogenous iliac crest bone graft; 2) fixation group: posterolateral arthrodesis L5-L6 with autogenous bone graft and interspinous fixation; 3) OP-1 group: in situ posterolateral L5-L6 arthrodesis using OP-1 and 4) combined OP-1 and fixation group. Radiographic fusion analysis was performed with computed tomography scans at 3 and 12 weeks after surgery. Decalcified histology was performed to assess tissue morphology and cellularity., Results: Minimal evidence of fusion was noted at 3 weeks with autograft or OP-1. By 12 weeks, all OP-1-treated animals had solid fusion, whereas no fusion was noted in autograft animals. The addition of fixation slightly increased radiographic fusion at 3 weeks in autograft and OP-1 groups but did not affect OP-1 animals at 12 weeks where all were fused. Decalcified histologic results confirmed the proliferative bone formation noted with OP-1 and the variable cellular response with autograft., Conclusions: The results of the present study suggest that the osteoinductive effect of OP-1 may be only minimally enhanced early in the bone healing process but does not appear to be affected in the long term by spinal fixation in the rabbit intertransverse fusion model. Fixation appeared to enhance early fusion in the autograft group.

Calcium phosphate-based bioceramics have revolutionized orthopedic and dental repair of damaged parts of the bone system. Among these materials, calcium phosphate-based cements, with hydraulic setting, stand out due to their biocompatibility and in situ hardening, which allow easy manipulation and adaptation to the shape and dimensions of bone defects. An investigation was made of the in vitro cytotoxic effect of calcium phosphate cement based on alpha-tricalcium phosphate, immersed for different lengths of time in simulated body fluid (SBF), based on the ISO-10993 "Biological Evaluation of Medical Devices" standard. The culture medium was Chinese hamster ovary (CHO) cells in contact with diluted cement extracts. The results revealed that the calcium phosphate cement used was cytotoxic and that the material's cytotoxicity decreased the longer the cement was immersed in SBF.

The expression of thrombomodulin after contact with CMW 1 bone cement extracts was studied in human umbilical vein endothelial cells. Cement extracts after 1 h and 7-day curing induced no significant variations in thrombomodulin antigen levels and in mRNA expression. Significant increase of thrombomodulin was observed when endothelial cells were treated with all-trans retinoic acid (ATRA). ATRA induced the increase of thrombomodulin also in cells incubated with cement extracts. These results suggest that CMW 1 bone cement does not impair the expression of thrombomodulin in endothelial cells.

There is increasing evidence that wear debris particles present in periprosthetic tissues have direct effects on osteoblasts. The nature of the cell response varies with the chemistry of the particle and the number of particles. Most studies have used Ti, Ti-6Al-4V, and ultrahigh molecular weight polyethylene (UHMWPE) particles since these materials are most frequently used in implants and as a result, these particles predominate in peri-prosthetic tissues. Ceramics have also been used successfully as load-bearing surfaces in implants for years, although it is unknown how wear debris from these surfaces may contribute to aseptic bone loss. Further, particles resulting from polymethylmethacrylate (PMMA) cements used for fixation may also be involved in aseptic loosening of implants, but how these particles may affect bone formation is unknown. In the present study, we examined whether aluminum oxide (Al2O3), zirconium oxide (ZrO2), and PMMA particles exert effects on osteoblast proliferation, phenotypic expression, and local factor production, and if so, whether the effects were specific to the particle type. ZrO2 particles were produced in a custom-made axial mixer in which ZrO2 containers were filled with ZrO2 bars and 95% ethanol and then rotated continuously at room temperature. PMMA particles were prepared in a ZrO2 roller mill. Al2O3 was produced and provided by Aesculap AG. Particles were endotoxin-free with equivalent circle diameters <3 microm; Al2O3 particles were significantly smaller than ZrO2 or PMMA particles. Particle suspensions were added to confluent cultures of MG63 osteoblast-like cells after diluting them 1:100, 1:10, and 1:1 with culture medium. Cells were incubated with the particles for 24 h. Transmission electron microscopy showed that MG63 cells phagocytosed Al2O3 particles and exhibited ultrastructural changes consistent with cytotoxicity. This was supported by biochemical changes as well. Proliferation, alkaline phosphatase activity, and TGF-beta1 levels were decreased. ZrO2 and PMMA particles increased proliferation and alkaline phosphatase specific activity. The effect of ZrO2 on alkaline phosphatase was targeted to matrix vesicles, the effect of PMMA was greater on the cells. All particles increased prostaglandin E2 production. These results show that Al2O3, ZrO2, and PMMA particles elicit direct effects on osteoblasts and that cell response depends on the particle type. None of the particles tested had the same effect as noted previously for UHMWPE: increased proliferation and decreased alkaline phosphatase. These results may indicate that the response of peri-prosthetic tissues to wear particles may be modulated by the relative contributions of the various particle types present.

The effect that three different radiopacifying agents, two of them inorganic (BaSO4, ZrO2) and one organic (an iodine containing monomer, IHQM) have on the static and dynamic mechanical properties of acrylic bone cements was studied. Compressive and tensile strength, fracture toughness and fatigue crack propagation were evaluated. The effect of the inorganic fillers depends on their size and morphology. In relation to the radiolucent cement, the addition of zirconium dioxide improved significantly the tensile strength, the fracture toughness and the fatigue crack propagation resistance. In contrast, the addition of barium sulphate produced a decrease of the tensile strength, but did not affect the fracture toughness and improved the crack propagation resistance. When the iodine containing monomer was used, although the tensile strength and the fracture toughness increased, the fatigue crack propagation resistance remained as low as it was for the radiolucent cement.

We investigated the effectiveness of an antioxidant agent, N-acetylcysteine (NAC), in suppressing macrophage activation and mediator release in response to particulate debris. Polymethylmethacrylate (PMMA) particle-stimulated monocyte-macrophages were cultured alone and with varying concentrations of NAC. Tumor necrosis factor alpha (TNFalpha) and interleukin-1beta (IL-1beta) expression in the resultant cultures were measured using enzyme-linked immunosorbant assays. The ultrastructural effect of treatment was also assessed by electron microscopy. Cell viability in the various cultures was measured to rule out an effect of cytotoxicity. NAC treatment reduced TNFalpha and IL-1beta expression by the monocyte-macrophages. Culturing with NAC was also associated with less ultrastructural activation of the monocytes. Furthermore, NAC was not associated with any adverse effect on cell viability in the concentrations used. Our findings demonstrate the effectiveness of the antioxidant N-acetylcysteine in suppressing the cell activation and TNFalpha release seen on exposure to wear debris. This represents a novel potential therapeutic method in the prevention or treatment of periprosthetic osteolysis.

Bone defects were created in rabbit medial femoral condyle in a model where subchondral bone was completely removed or about 2 mm of subchondral bone was maintained. Groups without augmentation and augmented with autogenous bone, polymethylmethacrylate, or calcium phosphate cement were sacrificed at 3, 12, and 24 weeks for evaluation of articular cartilage and observation of bone formation. In the model in which subchondral bone was completely removed, collapse of the subchondral bone together with exfoliation and prolapse of cartilage developed early in all cases. In the model in which 2 mm of subchondral bone was maintained, degeneration of articular cartilage developed at 12 weeks in the group augmented with polymethylmethacrylate, showing a significant difference when compared to the contralateral untreated control group. The group augmented with calcium phosphate cement did not demonstrate any evident difference from the control group. Mechanical properties after subchondral replacement did not differ between the groups augmented with polymethylmethacrylate and calcium phosphate cement, although calcium phosphate cement was considered histologically superior. Calcium phosphate cement was a reliable subchondral replacement material when the bone defect is adjacent to the articular cartilage., (Copyright 2001 John Wiley & Sons, Inc.)

Particulate prosthetic materials are often studied by adding them to monocytic cells in vitro and measuring the release of cytokines as an indicator of their inflammatory potential. Endotoxin is known to be a contaminant of particle preparations and also stimulates the release of cytokines. It is usual to use a proprietary endotoxin test to avoid erroneous results. Four different formulations of cement were found to be free from endotoxin using standard, gel-clot tests but stimulated different levels of release of cytokines from macrophages. These differences were explained when a more sensitive, kinetic endotoxin assay showed that release correlated with minor contamination with endotoxin. In a repeat experiment using cement particles with low or undetectable levels of endotoxin by kinetic assay, differences in the ability of the formulations to stimulate the release of cytokines were not seen. Endotoxin is adsorbed on to the surface of particles and it is this combination which stimulates increased release of cytokines. In both the above methods for determination of endotoxin, the water in which the particles had been soaked was examined rather than the particles directly. Further investigations showed that a kinetic assay directly on a particle suspension is the most sensitive method to measure contamination with endotoxin.

Three methacrylate-based bone cements used for the fixation of joint prostheses were evaluated: Sulfix-60 (Sulzer Orthopedic Inc., Baar, Switzerland). CMW1 (DePuy International Ltd., England). and CMW2 (DePuy International Ltd., England). The cements after polymerization were put in contact in vitro with platelet-rich plasma. Plasma, in contact only with siliconized glass, was used as a negative control. After contact, platelet number. beta-thromboglobulin (beta-TG), and transforming growth factor-beta1 (TGF-beta1) were determined. The Student's paired t test showed that the ccments induced no significant modifications of platelet number. CMWI and Sulfix-60 determined a significant increase in beta-TG compared with the negative control. All cements determined a significant increase in TGF-beta1. Significant differences were also seen in the levels of beta-TG and TGF-beta1 between cements with a content of benzoyl peroxide < 1 (Sulfix-60) and those with a content > 1 (CMW1 and CMW2). The cement with zirconium dioxide (Sulfix-60) produced higher levels of beta-TG and TGF-beta1, compared to those with barium sulphate (CMW1 and CMW2). In conclusion, all the cements induced the secretion of TGF-beta1 CMW1 and Sulfix-60 determined also a significant release of beta-TG. Platelet activation induced by the cements from one side could contribute to the pathogenesis of deep venous thrombosis, that often occurs after prosthetic implant and is caused also by other factors, including surgical trauma and venous stasis. From the other side, activated platelets can release growth factors favoring bone formation.

Purpose: The purpose of this study was to evaluate the short-term wound healing and repair at the tissue/cement interface with different hydroxyapatite cement formulations., Materials and Methods: Three groups of Sprague-Dawley rats were implanted with HAC for 3, 7, or 14 days. The medial femur-tibia sites on each leg of the animals were used to create four separate 1.5-mm defects, which were filled with HAC (Bone-Source, Howmedica-Leibinger, Inc.) mixed with either water (W) as a control, an accelerant (0.1 M sodium phosphate, SP), or a stabilizer (1% hydroxypropylmethyl cellulose, HPMC) as diluents. At the appropriate postimplantation times, the animals were euthanized and tissue-implant blocks were prepared for qualitative histopathologic evaluations., Results: Osseoconduction around and into the HAC was observed with all three cement formulations and the control (unfilled) site by day 7 and continued through day 14. The histologic staining did not reveal resorption lacunae or other cellular activities characteristic of osteoclast degradation of HAC. These results suggests that other processes, perhaps physical-chemical in nature, contributed to the initial degradation of the HAC following surgical placement. Structural stability of HAC was noted when using HPMC as the diluent. However, when either W or SP were employed, dissociation (washout) of the HAC into the surrounding tissue was noted., Conclusion: HAC is prone to in situ physical-chemical breakdown before the completion of the setting reaction. Diluents designed to stabilize or accelerate the HAC mixture appeared to improve the handling properties of the HAC without compromising the biological characteristics of the cement.

Injectable calcium phosphate hydraulic cements are known to have a high clinical potential in bone reconstruction for mini-invasive orthopaedic surgery, interventional radiology, and rheumatology. Previous in vivo experiments in rabbit have shown that the presence of beta-TCP granules in injectable bone cement help maintain the transient biomechanical function of the implanted bone and promote the formation of good-quality new bone. Histomorphometric analysis of two brushite hydraulic cement (BHC) mixtures selected from previous results (referred to in this work as BHC-A and BHC-B) was performed at three postoperative delays (0, 12, and 24 weeks): histomorphometric analysis of bone colonization within beta-TCP shows that, just before implantation, the beta-TCP granule area is significantly higher in BHC-B; the residual granule area decreases steadily over time in BHC-A, whereas it goes through a maximum of 30% at 12 weeks in BHC-B; the residual granule porosity increases steadily up to 35% in BHC-A, whereas it goes through a maximum of 35% at 12 weeks and decreases somewhat until 24 weeks in BHC-B. New bone formation within granules appears higher in BHC-A (58% Area) compared to BHC-B (38% area) at 12 weeks. At 24 weeks bone colonization levels off in both cements at about 50% area. Irrespective of the cement matrix composition, beta-TCP granules contribute actively to the conduction of new bone formation., (Copyright 2002 Wiley Periodicals, Inc. J Biomed Mater Res (Appl Biomater) 63: 413-417, 2002)

We conducted an experimental study using rabbits to determine the effect of injection of calcium phosphate cement (CPC) through a small cortical window on the torsional strength of the long bone. A drill hole created in the femoral shaft and medullary curettage before CPC injection simulates the clinical procedure for managing benign bone tumors. Torsional loading always produced a spiral fracture through the drill hole. The CPC-treated femurs, but not the polymethylmethacrylate-treated femurs, tolerated greater torsional loads and had greater energy absorption to failure at 24 h and 2 weeks postoperatively compared with the contralateral sham-operated femurs. These two parameters, however, were only 54% and 26%, respectively, of those in normal femurs at 24 h, but they were 71% and 51% of those in normal femurs at 12 weeks. Histologic sections demonstrated progressive covering of the drill hole by new bone in the CPC-treated specimens and invagination by newly formed cortical bone into the medullary cavity following sham operation. CPC increases the torsional strength of the long bones immediately after injection, although not sufficiently to preclude the need for external fixation in clinical applications. Better management of the cortical defect is needed to further improve torsional strength.

Seven acrylic bone cements were evaluated: Cemex Rx (Tecres S.p.a., Italy), Cemex Isoplastic (Tecres S.p.a., Italy), Zimmer Low Viscosity Cement (L.V.C., Zimmer, IN, USA), Zimmer bone cement - dough type (Zimmer, IN, USA), CMW (DePuy International Ltd., UK), Cerim LT (Cremascoli S.r.l., Italy), and Palacos (Merck, Wehreim, Germany). The cements after polymerization were put in contact in vitro with platelet-rich plasma. Plasma in contact only with siliconated glass was used as the negative control. After contact, platelet number, beta-thromboglobulin (beta-TG), and transforming growth factor-beta1 (TGF-beta1) were determined. The Wilcoxon signed rank test showed Palacos R and L.V.C. induced a significant decrease of platelet number compared with the negative control. All cements determined a significant increase in beta-TG. CMW 3, Palacos, L.V.C., and Zimmer dough type determined a significant increase in TGF-beta1 compared with the negative control.

C-C chemokines are soluble mediators that occur in a periprosthetic granuloma and influence recruitment, localization and activation of inflammatory cells. This study tested effects of titanium and polymethylmethacrylate (PMMA) particles on expression of selected C-C chemokines in cultured human fibroblasts. The C-C chemokines analyzed included monocyte chemoattractant protein-1. 2 (MCP-1. 2), monocyte inflammatory protein-1 alpha (MIP-1 alpha), and regulated on activation, normal T-cell expressed and secreted protein (RANTES). Interleukin-1 beta (IL-1 beta) served as a known stimulator of chemokine release while interleukin-6 (IL-6) expression served as a marker for fibroblast activation. Protein and mRNA signal levels were determined by ELISA and RT-PCR, respectively. The results demonstrated that exposure of fibroblasts to titanium and PMMA particles resulted in increased release of MCP-1 in a dose- and time-dependent manner. After 24 h, titanium particles maximally upregulated MCP-1 release 7-fold while PMMA particles increased MCP-1 levels 2-fold, when compared to unchallenged fibroblasts. MCP-2, MIP-1 alpha and RANTES levels remained unchanged following exposure of fibroblasts to titanium or PMMA particles at any concentration or time point tested. However, IL-1 beta stimulated release of MCP-1, MCP-2, and RANTES, but not MIP-1 alpha from the fibroblasts. IL-1 beta, not particles, exhibited the most prominent effect on MCP-1 mRNA levels. Increased release of MCP-1 from fibroblasts exposed to titanium and PMMA particles coincided with increased release of IL-6. This study suggests that release of chemoattractant factors from fibroblasts localized in periprosthetic membranes enhances the chronic inflammatory process leading to bone resorption and implant loosening.

Two rat models were used to characterize tissue-specific reactions to particles of bone-substitute materials: one for osteocompatibility in a healing tibial wound and the other in a heterotopic, subcutaneous site. Small, unicortical tibial wounds in rats healed spontaneously, beginning with the rapid proliferation of intramedullary woven bone. That temporary bone was resorbed by osteoclasts and finally, the cortical wound was healed with lamellar bone and the medullary space was repopulated with marrow. When various particulate materials were implanted into fresh wounds, three types of reactions were observed. (1) Demineralized bone powder (DBP) and non-resorbable calcium phosphate (nrCP) were incorporated into the reactive medullary and cortical bone. (2) Polymethylmetlhacrylate (PMMA) particles were surrounded with a fibrous layer, but did not impair bone healing. (3) Polyethylene (PE) shards and resorbable calcium phosphates (rCPs) were inflammatory and inhibited osseous repair. Subcutaneous sites showed osteoinductive, fibrotic, or inflammatory responses to these materials. Only DBP induced endochondral osteogenesis subcutaneously. The nrCP evoked a fibrous reaction. In contrast, rCPs, PMMA, and PE shards generated inflammatory reactions with each particle being surrounded by fibrous tissue and large multinucleated giant cells. In conclusion, only DBP showed osteoinductive as well as osteocompatible properties. The nrCP was osteocompatible. The rCPs stimulated various degrees of inflammatory responses. PMMA was osteocompatible and did not interfere with the bone healing process. PE was not osteocompatible and generated foreign body reactions in both sites. Use of the two sites distinguishes osteoinductive, osteocompatible, and inflammatory properties of particles of bone-substitute materials.

Periprosthetic membranes commonly observed at sites of total joint implant loosening exhibit abundant macrophages and particulate debris. Macrophages phagocytose orthopedic debris and release the pro-inflammatory mediators interleukin-1, interleukin-6, tumor necrosis factor-alpha, and prostaglandin E2. Populations of activated lymphocytes are often seen in periprosthetic membranes. These lymphocytes may modulate the monocyte/macrophage response to particulate debris and influence aseptic loosening. In addition, other immunologic agents, such as interleukin-10, are present in tissues harvested from the bone-implant interface of failed total joint arthroplasties. The present study examined the effects of interleukin-10 on polymethylmethacrylate (PMMA) particle challenged human monocyte/macrophages in vitro. Human monocyte/macrophages isolated from buffy coats of five healthy individuals were exposed to 1-10 microm PMMA particles. Interleukin-10 was added to the monocyte/macrophages with and without the addition of PMMA particles. Interleukin-10-induced alterations in monocyte/macrophage metabolism were determined measuring interleukin-6 and tumor necrosis factor-alpha release by the cells following exposure to PMMA particles. Exposure of the monocyte/macrophages to PMMA particles resulted in a dose-dependent release of interleukin-6 and tumor necrosis factor-alpha at 48 h. Interleukin-10 reduced the levels of interleukin-6 and tumor necrosis factor-alpha release by macrophages in response to PMMA particles in a dose-dependent manner. At 48 h, particle-induced interleukin-6 release was inhibited by 60 and 90% with 1.0 and 10.0 ng/ml treatments of interleukin-10, respectively. At 48 h, particle-induced tumor necrosis factor-alpha release was inhibited by 58 and 88% with 1.0 and 10.0 ng/ml treatments of interleukin-10, respectively. Interleukin-10 challenge alone did not significantly alter basal interleukin-6 or tumor necrosis factor-alpha release relative to control cultures. The data presented in this study demonstrate that the anti-inflammatory cytokine, interleukin-10, inhibits monocyte/macrophage release of the pro-inflammatory cytokines interleukin-6 and tumor necrosis factor-alpha in response to PMMA particle challenge in vitro.

Particulate Bioglass is a bioactive material used in the repair of periodontal defects. This material undergoes a series of surface reactions in an aqueous environment which lead to osseointegration. The aim of this study was to determine whether these reactions exerted an antibacterial effect on a range of oral bacteria. Streptococcus sanguis, Streptococcus mutans and Actinomyces viscosus were suspended in nutrient broth (NB), artificial saliva (AS) or Dulbecco's modified eagle medium plus 10% foetal calf serum (DMEM + 10%FCS), with or without particulate Bioglass. All bacteria showed reduced viability following exposure to Bioglass in all the media after 1 h. This antibacterial effect increased after 3 h. Porphyromonas gingivalis, Fusobacterium nucleatum, Prevotella intermedia and Actinobacillus actinomycetemcomitans were suspended in either BM broth or 40% horse serum (HS) in RPMI. A considerable reduction in viability was observed with all bacteria tested, in both media, compared to inert glass controls. In further experiments it was found that the viability of S. sanguis was significantly reduced following exposure to NB pre-incubated with Bioglass. Additionally, it was found that neutralisation of this highly alkaline solution eliminated the antibacterial effect. Moreover, a solution of NB and NaOH (of equivalent pH) exerted an antibacterial effect of similar magnitude to that of the solution pre-incubated with Bioglass. Thus, particulate Bioglass exerts an antibacterial effect on certain oral bacteria, possibly by virtue of the alkaline nature of its surface reactions. This may reduce bacterial colonisation of its surface in vivo.

Previously we developed a composite consisting of apatite and wollastonite containing glass-ceramic (AW-GC) powder and bisphenol-a-glycidyldimethacrylate (Bis-GMA)-based resin (designated AWC), and demonstrated that AWC showed direct contact with living bone. Another new composite consisting of mainly the delta-crystal phase of alumina bead powder and Bis-GMA-based resin (designated ABC) was developed. Although alumina ceramics are bioinert and a composite filled with the pure alpha-crystal phase of alumina powder (designated alphaALC) did not allow direct bone formation in vivo, ABC was shown to have excellent osteoconductivity. One purpose of this study was to investigate whether AW-GC powder in a composite promotes osteoblastic differentiation of rat bone marrow cells as AW-GC bulk did. Another purpose was to evaluate the effects of the delta-crystal phase of alumina powder in a composite on osteoblastic differentiation. In a cell culture with dexamethasone, alkaline phosphatase (AP) activity at both days 7 and 14, and the levels of osteocalcin mRNA and alpha1(I) collagen mRNA at day 14 and osteopontin mRNA at day 7, were highest on AWC, followed by ABC, and finally alphaALC. Scanning electron microscopy showed more abundant mineralized globules and a fibrous collagen matrix on AWC at day 14, followed by ABC. In a cell culture without dexamethasone, AP activity at both days 7 and 14, and the level of osteopontin mRNA at day 7, were higher on ABC than on any other composite, whereas osteocalcin mRNA could not be detected. These results indicate that AW-GC powder in a composite promotes osteoblastic differentiation of bone marrow cells intensively when supplemented with dexamethasone. The delta-crystal phase of alumina powder in a composite promotes greater osteoblastic differentiation than the alpha-crystal phase of alumina powder., (Copyright 2001 John Wiley & Sons, Inc.)

The effect of three methacrylate-based cements used for the fixation of joint prostheses on tissue-factor production by human umbilical vein endothelial cells was evaluated in vitro. The extracts in the culture medium of the cements were tested after 1-h and 7-day curing. The endothelial cells were incubated with the cement extracts for 4 h, and then the tissue factor was determined in cell lysates with both the recalcification time and enzyme immuno assay. The cements did not induce significant production of tissue factor and, therefore, did not activate the extrinsic pathway of coagulation within the limits of the mechanism considered.

A new bioactive bone cement (designated GBC), consisting of bioactive glass beads as an inorganic filler and polymethylmethacrylate (PMMA) as an organic matrix, has been developed. The purpose of the present study was to examine the effect of the amount of glass bead filler added to GBC on its mechanical and biological properties, and to decide the most suitable content of filler. Serial changes in GBC with time were also examined. The newly designed bioactive beads, consisting of MgO-CaO-SiO2-P2O5-CaF2 glass, were added to the cement in the proportions 30, 40, 50, 60, and 70 wt %. These cements were designated GBC30, GBC40, GBC50, GBC60, and GBC70, respectively. The compressive strength and the elastic modulus of bending of GBC increased as the glass bead content increased. The various types of GBC were packed into the intramedullar canals of rat tibiae to evaluate osteoconductivity, as determined by an affinity index calculated as the length of bone in direct contact with the cement expressed as a percentage of the total length of the cement surface. Rats were killed at 4 and 8 weeks after the operation and the affinity index was calculated for each type of GBC. Histologically, new bone had formed along the surface of all types of GBC within 4 weeks, even in GBC30 containing only 30 wt % of glass beads. At each time interval studied, there was a trend for the affinity index of GBC to increase as the glass bead filler content increased. There was no significant increase of affinity index between GBC60 and GBC70. The affinity indices for all types of GBC increased significantly with time up to 8 weeks. The handling properties of GBC were comparable to those of conventional PMMA bone cement. We conclude that when mechanical properties and osteoconductivity are both taken into consideration, GBC60 is the most suitable formulation; it shows excellent osteoconductivity and sufficient mechanical strength for clinical use.

We have investigated whether the particle-stimulated release of inflammatory cytokines from human primary macrophages in vitro was dependent upon the type of bone cement used. Particles of clinically relevant size were produced from Palacos R without radio-opacifier, Palacos R with BaSO4, Palacos R with ZrO2 and from CMW3 which contains BaSO4. All four preparations produced significantly greater release of tumour necrosis factor alpha, interleukin-6 and interleukin-1 beta than a negative control but there were no significant differences between them. The differences in the ability to stimulate bone resorption and in clinical performance between proprietary bone cements previously recorded are not explained by the release of the cytokines most commonly implicated in osteolysis.

The compatibility of a methacrylate-based bone cement (CMW 1, DePuy International Ltd, England) used for the fixation of joint prostheses was evaluated on plasma, an erythrocyte suspension and cultured human endothelial cells. The extract of the cement was tested, following 1 hour and 7 days of curing. After the contact in vitro of the extract with plasma, activated partial thromboplastin time, antithrombin III, thrombin-antithrombin complexes and fibrin degradation products were assayed. Hemolytic activity was tested by adding the cement extracts to a suspension of erythrocytes. After 4 hours of incubation at 37 degrees C, the hemoglobin concentration was determined on the supernatants by the colorimetric method. The effect of the cement on tissue factor and thrombomodulin production was evaluated on human umbilical vein endothelial cell cultures. Tissue factor was determined in cell lysates by enzyme immunoassay, following 4 hours' incubation of cultures with the cement extract. Thrombomodulin was assayed in cell lysates by enzyme immuno assay, after 24 hours' incubation with the cement extract. The response to all trans-retinoic acid (ATRA) was tested. The cement caused no significant modifications of the coagulation tests, had no hemolytic activity, did not determine tissue factor production and did not modify thrombomodulin, compared to the negative control. The response to stimulation with ATRA was similar to that of the negative control. We conclude that the cement extract does not affect the plasmatic phase of coagulation, has no effect on erythrocytes, does not induce the expression of procoagulant activity by endothelial cells and does not impair their antithrombotic property, within the limits of the tests performed.

Object: The aim of this in vitro study was to determine the influence of simulated postoperative neck movements on the stabilizing effect and subsidence of four different anterior cervical interbody fusion devices. Emphasis was placed on the relation between subsidence and spinal stability., Methods: The flexibility of 24 human cervical spine specimens was tested before and directly after being stabilized with a WING, BAK/C, AcroMed I/F cage, or with bone cement in standard flexibility tests under 50 N axial preload. Thereafter, 700 pure moment loading cycles (+/- 2 Nm) were applied in randomized directions to simulate physiological neck movements. Additional flexibility tests in combination with measurements of the subsidence depth were conducted after 50, 100, 200, 300, 500, and 700 loading cycles. In all four groups, simulated postoperative neck movements caused an increase of the range of motion (ROM) ranging from 0.4 to 3.1 degrees and of the neutral zone from 0.1 to 4.2 degrees. This increase in flexibility was most distinct in extension followed by flexion, lateral bending, and axial rotation. After cyclic loading, ROM tended to be lower in the group fitted with AcroMed cages (3.3 degrees in right lateral bending, 3.5 degrees in left axial rotation, 7.8 degrees in flexion, 8.3 degrees in extension) and in the group in which bone cement was applied (5.4 degrees, 2.5 degrees, 7.4 degrees, and 8.8 degrees, respectively) than in those fixed with the WING (6.3 degrees, 5.4 degrees, 9.7 degrees, and 6.9 degrees, respectively) and BAK cages (6.2 degrees, 4.5 degrees, 10.2 degrees, and 11.6 degrees, respectively)., Conclusions: Simulated repeated neck movements not only caused an increase of the flexibility but also subsidence of the implants into the adjacent vertebrae. The relation between flexibility increase and subsidence seemed to depend on the implant design: subsiding BAK/C cages partially supported stability whereas subsiding WING cages and AcroMed cages did not.

The present study examined the effects of in vitro challenge with an acrylic bone cement CMW 1 on the expression of transforming growth factor-beta 1 (TGF-beta 1) in human umbilical vein endothelial cells (HUVEC). The extracts in cell culture medium of the cements were tested, after 1 h and 7-day curing. Some cultures were also stimulated with interleukin-1 beta (IL-1 beta) or all-trans retinoic acid (ATRA). The expression of mRNA was evaluated by RT-PCR with specific primers. The release of TGF-beta 1 into the conditioned medium was evaluated by enzyme immunoassay. TGF-beta 1 mRNA was constitutively expressed by endothelial cells in the culture medium after 24 h. The incubation with the extracts of CMW 1, cured both for 1 h and 7 days, induced changes neither in mRNA expression, nor in the release of TGF-beta 1 into the conditioned medium, compared to the unstimulated cells. Even stimulation with ATRA, alone or added to the extracts at both curing times, affected neither mRNA expression nor TGF-beta 1 release, compared to the cells incubated with the cement alone or with the unstimulated cultures. The mRNA expression and the release were not changed by the stimulation with IL-1beta alone or added to the extract cured for 1 h. A significant decrease compared to the unstimulated cells was observed after the addition of IL-1 beta to the extract cured for 7 days. It was concluded that CMW 1 extract did not significantly modify TGF-beta 1 expression after 1-h curing, or after 7-day curing. Incubation with CMW 1 added with ATRA did not produce any changes in TGF-beta 1 synthesis. Incubation with cement extract after 7-day curing added with IL-beta 1 produced a significant reduction in TGF-beta 1 release.

Study Design: A biomechanical in vitro subsidence test of different cervical interbody fusion devices was performed using a new testing protocol that simulates physiologic conditions., Objectives: To investigate the effect of simulated postoperative neck movements on the subsidence of the new WING cervical interbody fusion cage in comparison with two other cages and bone cement., Summary of Background Data: Cervical interbody fusion cages sometimes cause complications because of subsidence into the adjacent vertebrae with collapse of the intervertebral space. Complications such as cage dislocation or nonunion with instability also have been reported. To prevent such complications, the new WING cervical interbody fusion cage (Medinorm AG, Quierschied, Germany) has been developed. Its area of contact with the adjacent vertebrae is supposed to be large enough to resist excessive subsidence and small enough to prevent stress protection of the tissue growing in the cage., Methods: In this study, 24 human cervical spine specimens were tested after stabilization with either a WING, BAK/C, AcroMed I/F cage or bone cement. Then, in a new testing protocol, 700 pure-moment loading cycles (+/-2 Nm) were applied in randomized directions (lateral bending, flexion-extension, and axial rotation alone or in combination with each other) to simulate the patient's neck movements during the first few postoperative days. Measurements of the subsidence depth (total height loss) in combination with flexibility tests (+/-2.5 Nm) were performed before cyclic loading and after 50, 100, 200, 300, 500, and 700 loading cycles., Results: Cyclic loading caused subsidence in all four device groups, most distinct with BAK/C-cages (1.63 mm after 700 loading cycles) followed by the new WING (0.90 mm) and the AcroMed (0.82 mm) cages. No statistically significant difference could be found among the three cage designs. However, all three cage types showed a significantly higher subsidence depth than bone cement (0.48 mm;P = 0.023 between each of the three cage-types and bone cement). A moderate correlation between bone mineral density and subsidence depth could be found only in the BAK/C group (r2 = 0.495). A large subsidence depth after 700 loading cycles was associated with a large flexibility increase in the WING (r2 = 0.786) and AcroMed groups (r2 = 0.21), but with a small flexibility increase in the BAK/C group (r2 = 0.58)., Conclusions: Postoperative neck movements caused subsidence in all cervical interbody implant types. The new WING cage and the AcroMed cage seemed to have a better resistance against subsidence than the BAK/C cage. However, all three cage types had a significantly higher subsidence tendency than bone cement.

Selective drug delivery to any organ becomes very important in certain diseases and clinical manifestations, especially when the drug affects other exposed tissues adversely. The importance of selective drug action is still further increased when the affected part is poorly perfused. Although a network of blood vessels is present throughout skeletal tissue (or bone), it is not sufficient for immediate delivery of drugs to the desired site of action in the tissue and in sufficient amounts over appropriate time periods. Hence, selective drug delivery to the skeletal system has remained a great challenge to pharmaceutical scientists over the years. However, in the recent past, attention has been focused on the importance of skeletal drug delivery and the first Skeletal Drug Delivery System (SDDS) was introduced by Bucholz and Engelbrecht in 1970 for the delivery of drugs to skeletal tissues at a high concentration to achieve desirable therapeutic effects. SDDS is used to deliver the drug directly to skeletal tissue through self-setting cement, which also acts as a bone filler, thereby improving the therapeutic effectiveness of drugs in bone diseases. The present review highlights the applicability of various materials as bone fillers for the purpose of skeletal drug delivery.

The authors evaluated the ability of bone cement to modify the profile of pro-inflammatory cytokines secreted by the immune cells. Peripheral blood mononuclear cells (PBMC) collected from healthy individuals were cultured with cement extracts and tested to assess the release of IL-1beta, TNFalpha, GM-CSF and IL-6 in both unstimulated and PHA-stimulated PBMC. The cytokine release of unstimulated PBMC was very poor, and in particular the IL-1beta was undetectable: the addition of cement extract increased both TNFalpha and GM-CSF release and decreased IL-6, sometimes significantly. The most recurrent observation in PHA-stimulated PBMCs exposed to bone cement extract was the increase in both IL-1beta and IL-6 release, while both the mean concentration and the index of release of TNFalpha and GM-CSF were changeable. In conclusion our results showed that leachable components of some bone cements can induce in vitro the release of pro-inflammatory cytokines which are known to be involved in the bone resorption associated with aseptic loosening of hip prostheses. These findings allowed us to identify materials endowed with the highest inflammatory power.

A new bioactive bone cement (designated GBC) consisting of polymethyl methacrylate (PMMA) as an organic matrix and bioactive glass beads as an inorganic filler has been developed. The bioactive beads, consisting of MgO-CaO-SiO(2)-P(2)O(5)-CaF(2) glass, have been newly designed, and a novel PMMA powder was selected. The purpose of the present study was to compare this new bone cement GBC's mechanical properties in vitro and its osteoconductivity in vivo with cements consisting of the same matrix as GBC and either apatite- and wollastonite-containing glass-ceramic (AW-GC) powder (designated AWC) or sintered hydroxyapatite (HA) powder (HAC). Each filler added to the cements amounted to 70 wt %. The bending strength of GBC was significantly higher than that of AWC and HAC (p < 0.0001). Cements were packed into intramedullar canals of rat tibiae in order to evaluate osteoconductivity as determined by an affinity index. Rats were sacrificed at 2, 4, and 8 weeks after operation. An affinity index, which equaled the length of bone in direct contact with the cement expressed as a percentage of the total length of the cement surface, was calculated for each cement. At each time interval studied, GBC showed a significantly higher affinity index than AWC or HAC up to 8 weeks after implantation (p < 0.03). The value for GBC increased significantly with time up to 8 weeks (p < 0.006). The handling property of GBC was comparable with that of PMMA bone cement. Our study revealed that the higher osteoconductivity of GBC was due to the higher bioactivity of the bioactive glass beads at the cement surface and the lower solubility of the new PMMA powder to MMA monomer. In addition, it was found that the smaller spherical shape and glassy phase of the glass beads gave GBC strong enough mechanical properties to be useful under weight-bearing conditions. GBC shows promise as an alternative with improved properties to the conventionally used PMMA bone cement., (Copyright 2000 John Wiley & Sons, Inc.)

This study analysed MIF mRNA and protein expression in human macrophages exposed in vitro to polymethylmethacryate and titanium alloy particles. MIF levels released from macrophages without exposure to particles were in the range of 2-4 ng/ml. Exposure of macrophages to particles as demonstrated increased MIF release at 0. 075%-0.225% v/v particle concentration, which was maximal at 12-24 h. MIF mRNA signal levels in cells with and without particles at a concentration of 0.075% showed no significant differences in a time course experiment. The profile of MIF release in response to increasing particle concentration coincided with increased release of lactate dehydrogenase. The viability of the cells was unchanged by the addition of particles as determined by 3H-thymidine uptake. These data suggest that MIF expression may represent an independent macrophage response to locally high particle concentrations., (Copyright 2000 Academic Press.)