S53P4 bioactive glass scaffolds induce BMP expression and integrative bone formation in a critical-sized diaphysis defect treated with a single-staged induced membrane technique.

Critical-sized diaphysis defects are complicated by inherent sub-optimal healing conditions. The two-staged induced membrane technique has been used to treat these challenging defects since the 1980's. It involves temporary implantation of a membrane-inducing spacer and subsequent bone graft defect filling. A single-staged, graft-independent technique would reduce both socio-economic costs and patient morbidity. Our aim was to enable such single-staged approach through development of a strong bioactive glass scaffold that could replace both the spacer and the graft filling. We constructed amorphous porous scaffolds of the clinically used bioactive glass S53P4 and evaluated them in vivo using a critical-sized defect model in the weight-bearing femur diaphysis of New Zealand White rabbits. S53P4 scaffolds and standard polymethylmethacrylate spacers were implanted for 2, 4, and 8 weeks. Induced membranes were confirmed histologically, and their osteostimulative activity was evaluated through RT-qPCR of bone morphogenic protein 2, 4, and 7 (BMPs). Bone formation and osseointegration were examined using histology, scanning electron microscopy, energy-dispersive X-ray analysis, and micro-computed tomography imaging. Scaffold integration, defect union and osteosynthesis were assessed manually and with X-ray projections. We demonstrated that S53P4 scaffolds induce osteostimulative membranes and produce osseointegrative new bone formation throughout the scaffolds. We also demonstrated successful stable scaffold integration with early defect union at 8 weeks postoperative in critical-sized segmental diaphyseal defects with implanted sintered amorphous S53P4 scaffolds. This study presents important considerations for future research and the potential of the S53P4 bioactive glass as a bone substitute in large diaphyseal defects. STATEMENT OF SIGNIFICANCE: Surgical management of critical-sized diaphyseal defects involves multiple challenges, and up to 10% result in delayed or non-union. The two-staged induced membrane technique is successfully used to treat these defects, but it is limited by the need of several procedures and bone graft. Repeated procedures increase costs and morbidity, while grafts are subject to donor-site complications and scarce availability. To transform this two-staged technique into one graft-independent procedure, we developed amorphous porous scaffolds sintered from the clinically used bioactive glass S53P4. This work constitutes the first evaluation of such scaffolds in vivo in a critical-sized diaphyseal defect in the weight-bearing rabbit femur. We provide important knowledge and prospects for future development of sintered S53P4 scaffolds as a bone substitute.

Sintered S53P4 bioactive glass scaffolds have anti-inflammatory properties and stimulate osteogenesis in vitro

Bioactive glasses (BAG) are used as bone-graft substitutes in orthopaedic surgery. A specific BAG scaffold was developed by sintering BAG-S53P4 granules. It is hypothesised that this scaffold can be used as a bone substitute to fill bone defects and induce a bioactive membrane (IM) around the defect site. Beyond providing the scaffold increased mechanical strength, that the initial inflammatory reaction and subsequent IM formation can be enhanced by coating the scaffolds with poly(DL-lactide-co-glycolide) (PLGA) is also hypothesised. To study the immunomodulatory effects, BAG-S53P4 (± PLGA) scaffolds were placed on monolayers of primary human macrophage cultures and the production of various pro- and anti-inflammatory cytokines was assessed using reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) and ELISA. To study the osteogenic effects, BAG-S53P4 (± PLGA) scaffolds were cultured with rabbit mesenchymal stem cells and osteogenic differentiation was evaluated by RT-qPCR and matrix mineralisation assays. The scaffold ion release was quantified and the BAG surface reactivity visualised. Furthermore, the pH of culture media was measured. BAG-S53P4 scaffolds had both anti-inflammatory and osteogenic properties that were likely attributable to alkalinisation of the media and ion release from the scaffold. pH change, ion release, and immunomodulatory properties of the scaffold could be modulated by the PLGA coating. Contrary to the hypothesis, the coating functioned by attenuating the BAG surface reactions and subsequent anti-inflammatory properties, rather than inducing an elevated inflammatory response compared to BAG-S53P4 alone. These results further validated the use of BAG-S53P4 (± PLGA) scaffolds as bone substitutes and indicate that scaffold properties can be tailored to a specific clinical need.

Reconstruction of Vertebral Bone Defects using an Expandable Replacement Device and Bioactive Glass S53P4 in the Treatment of Vertebral Osteomyelitis: Three Patients and Three Pathogens.

BACKGROUND AND AIMS: Bioactive glass S53P4 is an antibacterial bone substitute with bone-bonding and osteostimulative properties. The bone substitute has been successfully used clinically in spine; trauma; orthopedic; ear, nose, and throat; and cranio-maxillofacial surgeries. Bioactive glass S53P4 significantly reduces the amount of bacteria in vitro and possesses the capacity to kill both planktonic bacteria and bacteria in biofilm. Three patients with severe spondylodiscitis caused by Mycobacterium tuberculosis, Candida tropicalis, or Staphylococcus aureus were operatively treated due to failed conservative treatment. The vertebral defects were reconstructed using bioactive glass S53P4 and an expandable replacement device. MATERIAL AND METHODS: Decompression and a posterolateral spondylodesis, using transpedicular fixation, were performed posteriorly in combination with an anterior decompression and reconstruction using an expandable vertebral body replacement device. For patients 1 and 2, the expander was covered with bioactive glass S53P4 only, and for patient 3, the glass was mixed with autograft bone. RESULTS: The patients healed well with complete neurological recovery. Fusion was observed for all patients. The total follow-up was 4 years for patient 1, 1 year and 8 months for patient 2, and 2 years and 2 months for patient 3. No relapses or complications were observed. CONCLUSION: The antibacterial properties of bioactive glass S53P4 also make it a suitable bone substitute in the treatment of severe spondylodiscitis.

Posterolateral spondylodesis using bioactive glass S53P4 and autogenous bone in instrumented unstable lumbar spine burst fractures. A prospective 10-year follow-up study.

BACKGROUND AND AIMS: A prospective long-term follow-up study of bioactive glass (BAG)-S53P4 and autogenous bone (AB) used as bone graft substitutes for posterolateral spondylodesis in treatment of unstable lumbar spine burst fractures during 1996-1998 was conducted. MATERIAL AND METHODS: The lumbar fractures were fixed using posterior USS instrumentation. BAG was implanted on the left side of the fusion-bed and AB on the right side. The operative outcome was evaluated on X-rays and CT scans, and a clinical examination was also performed. RESULTS: The Oswestry score was excellent, and the mean pain score 1. The mean compression rate of the injured vertebral body was 25%. A solid bony fusion was seen on CT scans on the AB side in all patients and on the BAG side in five patients, and a partial fusion in five patients, resulting in a total fusion-rate of 71% of all fused segments in the BAG group. CONCLUSIONS: Our long-term results show that BAG-S54P4 bone graft material is safe to be used as a bone graft extender in spine surgery.

Bioactive glass S53P4 as bone graft substitute in treatment of osteomyelitis.

Bioactive glass (BAG)-S53P4 is an osteoconductive bone substitute with proven antibacterial and bone bonding properties. In a multicentre study 11 patients with verified chronic osteomyelitis in the lower extremity and the spine were treated with BAG-S53P4 as a bone substitute. The cavitary bone defect and the surrounding of a spinal implant were filled with BAG-S53P4. The most common pathogen causing the infection was Staphylococcus aureus. The mean follow-up was 24 months (range 10-38). BAG-S53P4 was well tolerated. Nine patients healed without complications. One patient who achieved good bone formation sustained a superficial wound infection due to vascular problems in the muscle flap, and one patient had an infection due to a deep haematoma. This study shows that BAG-S53P4 is a good and well-tolerated bone substitute, and can be used in treatment of osteomyelitis with good primary results.

Complications in the upper extremity following intra-arterial drug abuse.

A retrospective study of drug abuse patients who developed arterial and venous complications in the upper extremity during 2002-2006 was performed. Twenty-two patients were admitted to hospital on 24 occasions over this period for treatment by our hand clinic. The drug most frequently causing complications was midazolam. The predominant clinical findings were increasing pain and loss of sensitivity in the hand, followed by oedema, cyanosis and marbling of the skin. Treatments included brachial block anaesthesia, low molecular weight heparin, embolectomy and fasciotomies. Despite these measures, amputations, mainly of the fingertips, were necessary in 15 patients. Complications in the upper extremity after self-injection by drug addicts are increasing; information and preventive procedures to minimize these complications are important and demanding tasks for health care bodies.

Treatment of a recurrent aneurysmal bone cyst with bioactive glass in a child allows for good bone remodelling and growth.

A recurrent aneurysmal bone cyst of the proximal phalanx of the index finger of a three-year-old child, treated with bioactive glass, is presented. Over a two-year follow-up, the treated area appeared dense on X-rays. At two years, no cavity was observed and the homogenous region resembled normal trabecular bone. The phalanx had grown in length and remodelled to an almost normal shape.

Long-term evaluation of blood silicon and ostecalcin in operatively treated patients with benign bone tumors using bioactive glass and autogenous bone.

In a study on 25 patients with verified benign bone tumors, bioactive glass (BG) and autogenous bone (AB) were used as bone-graft substitutes. The patients were randomized into two groups according to the filling material. Blood samples were taken both preoperatively, at 2 weeks, and 3, 8, 12, 24, and 36 months postoperatively, for evaluation of silicon concentration in blood. In the determination, direct current plasma atomic emission spectroscopy was used. No significant difference in blood silicon concentration between the BG group or the AB group could statistically be observed (p = 0.5400), and neither did the size of the bone tumor (p = 0.4259) nor the follow-up time affect the results (p = 0.2094). Concentration of osteocalcin in blood was significantly higher for large cysts (p < 0.0001). The filler material (BG or AB) did not affect the osteocalcin concentration level in blood.

Granule size and composition of bioactive glasses affect osteoconduction in rabbit.

Bioactive glass granules of three different compositions, regarding particularly Si- and Al- content (S53P4, S59.7P2.5, S52P3) and of two different granule sizes (200-250 microm and 630-800 microm) were implanted for 4 and 8 weeks in the distal part of rabbit femur. The effect of glass composition and granule size on bone formation was studied. The results were evaluated using histology, computerized histomorphometry, scanning electron microscopy and energy dispersive X-ray analysis, and used for mathematical description of bone formation. The results showed that both the composition of the glass and the granule size of the granules, have influence on bone growth from the surrounding tissue. Glass S53P4, which from previous observations is known to be an effective bioactive glass and widely used in the Biomaterial Project of Turku, Finland, showed bone bonding and increasing bone growth between the granules. Glass S59.7P2.5 which due to its high Si-content should be inert, showed bone bonding. At 4 weeks the bone growth was significantly more abundant in bone defects filled with large granules (630-800 microm) than in defects filled with small granules (200-250 microm). Glass S52P3 with an alumina content of 3 wt %, showed good bone conduction, possibly even bone bonding for granules of 630-800 microm size. Granules of 200-250 microm with a high alumina content at the surface of the reaction layer, showed hardly any bone contact at all. This data, therefore, gives new information concerning bone bonding and osteoconduction of bioactive glasses with a high silica or alumina content.

Bioactive glass as bone-graft substitute for posterior spinal fusion in rabbit.

Bioactive glass S53P4 and autogenous bone were studied as bone graft materials for spinal fusion in a rabbit model. Sixteen rabbits underwent surgery by a dorsal approach. A bioactive glass, a combination of bioactive glass and autogenous bone (70/30 vol%), and autogenous bone were implanted at two thoracolumbar vertebraes for 4 and 12 weeks. The volume, consolidation to vertebrae, and fusion of the graft material were evaluated with plain-film radiology, computed tomography (CT) and bone-mineral density measurements, and compared with histomorphometrical measurements. Radiological consolidation by CT of bone graft to underlying vertebrae at 12 weeks was observable in all groups. This was histologically confirmed as bone was growing from the vertebrae into the graft material. Radiologic fusion of vertebraes was, at 12 weeks, observable in all groups in 50--75% of the cases. The radiologic fusion seen at the CT scans could, however, not be confirmed by histology in any of the three groups. Significant differences for graft material and observation period with the use of bone-mineral density measurements (Hounsfield units) were also observable, with the highest measured values for the bioactive glass group and the lowest for the autogenous bone group. The results indicate that bioactive glass have potential as bone-graft material in spinal fusion. The reliability of radiologic evaluation methods in spinal surgery using bone substitutes is also questioned and discussed.

Immunoglobulin enhances the bioactive-glass-induced chemiluminescence response of human polymorphonuclear leukocytes.

Bioactive glasses are bone substitutes that chemically bind to bone. Implanted materials always elicit a response from surrounding tissues and thereby can activate inflammatory cells, with subsequent release of biomaterial and tissue-damaging agents. Bioactive glasses can activate polymorphonuclear leukocytes (PMNL) and induce a release of reactive oxygen metabolites (ROM). Adsorption of proteins on the surface of the implanted material may influence the subsequent inflammatory cell response. The effect of Sandoglobulin(R) (SG) and albumin on the ROM release by PMNLs induced by a bioactive glass was studied by a chemiluminescence (CL) assay. An enhanced effect for SG and inhibitory effect for albumin on the CL response of PMNLs was observable. The CL response of the PMNLs was dependent on the incubation time of the glass in solution.

Tissue response to bioactive glass and autogenous bone in the rabbit spine.

Bioactive glass S53P4 and autogenous bone were used as bone graft materials in an experimental rabbit model for spinal fusion. The study focused on differences in bone formation using bioactive glass and autogenous bone as bone graft materials. Bioactive glass, a mixture of bioactive glass and autogenous bone or autogenous bone was implanted for 4 and 12 weeks at the thoracolumbar level. Undecalcified sections were prepared for histological and histomorphometric evaluation. New bone formation was seen in all implanted areas, with the bone growing from the surface of the vertebrae enclosing both glass and autogenous bone in the bone fusion mass. During the observation period, the measured amount of bone remained at the same level in the autograft group, while in the glass and the glass/autograft bone groups it increased. By 12 weeks, no significant difference in bone formation between the three groups was observable. The bone formation in two selected standardized areas at 12 weeks was 21 and 24% in the glass group, 23 and 28% in the glass/autograft bone group and 27 and 26% in the autograft bone group. We consider bioactive glass as a potential bone graft material in experimental spinal fusion.

Bioactive glasses induce chemiluminescence by human polymorphonuclear leukocytes.

The effect of bioactive glasses on human polymorphonuclear leukocytes (PMNLs) were studied in vitro by a chemiluminescence (CL) assay. Eight different glasses were chosen. All glasses induced a rapid CL response by human PMNLs, which proved to be dose dependent. The CL response also seemed to depend on the durability of the glasses. The least durable glass caused the highest CL response, and highly durable glasses caused only low CL responses by the cells.