CaO-B2O3-SiO2 glass fibers for wound healing.

It was reported by Jung and Day in 2011 that a cotton-like glass fiber pad made of borate glass 13-93B3 demonstrated a remarkable wound healing effect. It was approved for sale as a novel wound dressing in the management of acute and chronic wounds in 2016. However, the detailed mechanism of its wound healing effect has not been reported. In the present study, glass fibers of different composition in the system CaO-B2O3-SiO2 were prepared and their in vitro properties investigated to determine the role of the constituent components in wound healing. Fine glass fibers that were 0.6-2.0 µm in diameter were obtained by a melt blown method. However, these fibers were accompanied by small glass beads because of the low viscosity of the glass melts. 13-93B3 glass released an appreciable amount of borate and calcium ions into simulated body fluid (SBF). The amounts of these released ions decreased with partial replacement of the B2O3 in 13-93B3 with SiO2. The addition of large amounts of the borate and calcium ions into the culture medium decreased the viability of the L929 fibroblasts. Partial replacement of the B2O3 in 13-93B3 with SiO2 induced the formation of an apatite-like phase amenable to the adsorption of biological components on its surface in SBF. The wound healing effect of these glass fibers of different composition is worth examining in future animal experiments.

Apatite-forming PEEK with TiO2 surface layer coating.

Polyetheretherketone (PEEK) is widely used in orthopedic implants, such as spinal fusion devices, because of its moderate elastic modulus, as well as relatively high mechanical strength. However, it does not bond to living bone, and hence it needs autograft to be fixed to the bone. In this study, we attempted to add bone-bonding properties to PEEK by coating with TiO2 synthesized by the sol-gel process. When a TiO2 sol solution consisting of titanium isopropoxide, water, ethanol, and nitric acid was deposited on a PEEK substrate without any pretreatment, the formed TiO2 gel layer was easily peeled off after subsequent treatments. However, when the same solution was deposited on PEEK that was preliminarily subjected to UV or O2 plasma treatment, the deposited TiO2 gel layer strongly adhered to the substrate even after subsequent treatments. The strong adhesion was attributed to the interaction among the C-O, C=O, and O-C=O groups on the PEEK owing to the UV or O2 plasma treatment and the Ti-O bond of the TiO2 gel. Apatite did not form on the as-formed TiO2 gel layer in a simulated body fluid (SBF) even within 3 days; however, apatite formed after soaking in 0.1 M HCl solution at 80 °C for 24 h. This apatite formation was attributed to positive surface charge of the TiO2 gel layer induced by the acid treatment. The PEEK with the TiO2 gel layer coating formed by the proposed process is expected to bond to living bone, because a positively charged titanium oxide which facilitates the formation of apatite in SBF within a short period is known to bond to living bone.

In vivo study of the early bone-bonding ability of Ti meshes formed with calcium titanate via chemical treatments.

Alkali and heat (AH) treatment forming sodium titanate has been shown to connect bioinert Ti metal and bone tissue. Artificial joints treated with this method have achieved extensive clinical application. Recently a new chemical treatment of Alkali-Calcium-Heat-Water (ACaHW) treatment forming calcium titanate was proposed. Notably, the apatite-forming ability of this treatment is greater than that of AH treatment, as verified in vitro. However, the early bone-bonding abilities of the two treatments have not been compared in vivo. To simulate clinical application, we treated a commercially pure Ti (Cp-Ti) mesh implant with AH or ACaHW. Then, using mechanical and histological methods, we compared the bone-bonding abilities of the two treatments early during the implantation process (2-4 weeks); untreated Cp-Ti mesh was used as a control. Because the mesh structure might influence bone-bonding ability, we compared these bonding abilities with values obtained at 4 and 8 weeks using a Cp-Ti implant with a plate structure. In the mesh group, histological comparisons at 2 and 3 weeks indicated that ACaHW treatment resulted in a bone-bonding ability similar to that of AH treatment; ACaHW exhibited a greater bonding ability than AH at 4 weeks. However, in tests of the plate group at later time points, such differences were not apparent. The results obtained here indicate that during the early stage of embedment, ACaHW treatment of Cp-Ti mesh implants yields a higher bone-bonding ability than AH treatment, thus providing a positive reference for future clinical applications.

A unified in vitro evaluation for apatite-forming ability of bioactive glasses and their variants.

The aim of this study was to propose and validate a new unified method for testing dissolution rates of bioactive glasses and their variants, and the formation of calcium phosphate layer formation on their surface, which is an indicator of bioactivity. At present, comparison in the literature is difficult as many groups use different testing protocols. An ISO standard covers the use of simulated body fluid on standard shape materials but it does not take into account that bioactive glasses can have very different specific surface areas, as for glass powders. Validation of the proposed modified test was through round robin testing and comparison to the ISO standard where appropriate. The proposed test uses fixed mass per solution volume ratio and agitated solution. The round robin study showed differences in hydroxyapatite nucleation on glasses of different composition and between glasses of the same composition but different particle size. The results were reproducible between research facilities. Researchers should use this method when testing new glasses, or their variants, to enable comparison between the literature in the future.

Evaluation of bioactivity of alkali- and heat-treated titanium using fluorescent mouse osteoblasts.

Stimulation of osteoblast proliferation and differentiation is important for the in vivo bone-bonding ability of biomaterials. Previous in vitro studies have used biochemical assays to analyze osteoblast-specific gene expression in cultured osteoblasts. In this study, we generated transgenic mice harboring a monomeric red fluorescent protein 1 transgene under the control of a 2.3-kb fragment of the Col1a1 promoter, which is active specifically in osteoblasts and osteocytes. We established a fluorescent primary osteoblast culture system to allow noninvasive observation of osteoblast proliferation and differentiation on opaque materials in vitro. We used this system to evaluate alkali- and heat-treated titanium, which has a strong bone-bonding ability in vivo, and we observed a rapid increase in fluorescence intensity and characteristic multifocal nodule formation. A cell proliferation assay and RT-PCR to examine osteoblast-specific gene expression showed increased osteoblast proliferation and differentiation consistent with the fluorescence observations. This mouse model allowed us to use fluorescence intensity to visualize and quantify in vivo newly formed bone around implanted materials in femurs. The use of these fluorescent osteoblasts is a promising method for simple screening of the bone-bonding ability of new materials.

Antibacterial and bioactive calcium titanate layers formed on Ti metal and its alloys.

An antibacterial and bioactive titanium (Ti)-based material was developed for use as a bone substitute under load-bearing conditions. As previously reported, Ti metal was successively subjected to NaOH, CaCl2, heat, and water treatments to form a calcium-deficient calcium titanate layer on its surface. When placed in a simulated body fluid (SBF), this bioactive Ti formed an apatite layer on its surface and tightly bonded to bones in the body. To address concerns regarding deep infection during orthopedic surgery, Ag(+) ions were incorporated on the surface of this bioactive Ti metal to impart antibacterial properties. Ti metal was first soaked in a 5 M NaOH solution to form a 1 µm-thick sodium hydrogen titanate layer on the surface and then in a 100 mM CaCl2 solution to form a calcium hydrogen titanate layer via replacement of the Na(+) ions with Ca(2+) ions. The Ti material was subsequently heated at 600 °C for 1 h to transform the calcium hydrogen titanate into calcium titanate. This heat-treated titanium metal was then soaked in 0.01-10 mM AgNO3 solutions at 80 °C for 24 h. As a result, 0.1-0.82 at.% Ag(+) ions and a small amount of H3O(+) ions were incorporated into the surface calcium titanate layers. The resultant products formed apatite on their surface in an SBF, released 0.35-3.24 ppm Ag(+) ion into the fetal bovine serum within 24 h, and exhibited a strong antibacterial effect against Staphylococcus aureus. These results suggest that the present Ti metals should exhibit strong antibacterial properties in the living body in addition to tightly bonding to the surrounding bone through the apatite layer that forms on their surfaces in the body.

Bone bonding ability of a chemically and thermally treated low elastic modulus Ti alloy: gum metal.

The gum metal with composition Ti-36Nb-2Ta-3Zr-0.3O, is free from cytotoxic elements and exhibits a low elastic modulus as well as high mechanical strength. We have previously demonstrated that this gum metal, once subjected to a series of surface treatments--immersion in 1 M NaOH (alkali treatment) and then 100 mM CaCl2, before heating at 700 °C (sample: ACaH-GM), with an optional final hot water immersion (sample: ACaHW-GM)--has apatite-forming ability in simulated body fluid. To confirm the in vivo bioactivity of these treated alloys, failure loads between implants and bone at 4, 8, 16, and 26 weeks after implantation in rabbits' tibiae were measured for untreated gum metal (UT-GM), ACaH-GM and ACaHW-GM, as well as pure titanium plates after alkali and heat treatment (AH-Ti). The ACaH-GM and UT-GM plates showed almost no bonding, whereas ACaHW-GM and AH-Ti plates showed successful bonding by 4 weeks, and their failure loads subsequently increased with time. The histological findings showed a large amount of new bone in contact with the surface of ACaHW-GM and AH-Ti plates, suggesting that the ACaHW treatment could impart bone-bonding bioactivity to a gum metal in vivo. Thus, with this improved bioactive treatment, these advantageous gum metals become useful candidates for orthopedic and dental devices.

Effect of Ca contamination on apatite formation in a Ti metal subjected to NaOH and heat treatments.

It has long been known that titanium (Ti) metal bonds to living bone through an apatite layer formed on its surface in the living body after it had previously been subjected to NaOH and heat treatments and as a result had formed sodium titanate on its surface. These treatments were applied to a porous Ti metal layer on a total hip joint and the resultant joint has been in clinical use since 2007. It has been also demonstrated that the apatite formation on the treated Ti metal in the living body also occurred in an acelullar simulated body fluid (SBF) with ion concentrations nearly equal to those of the human blood plasma, and hence bone-bonding ability of the treated Ti metal can be evaluated using SBF in vitro. However, it was recently found that certain Ti metals subjected to the same NaOH and heat treatments display apatite formation in SBF which is decreased with the increasing volume of the NaOH solution used in some cases. This indicates that bone-bonding ability of the treated Ti metal varies with the volume of the NaOH solution used. In the present study, this phenomenon was systematically investigated using commercial NaOH reagents and is considered in terms of the structure and composition of the surface layers of the treated Ti metals. It was found that a larger amount of the calcium contamination in the NaOH reagent is concentrated on the surface of the Ti metal during the NaOH treatment with an increasing volume of the NaOH solution, and that this inhibited apatite formation on the Ti metal in SBF by suppressing Na ion release from the sodium titanate into the surrounding fluid. Even a Ca contamination level of 0.0005 % of the NaOH reagent was sufficient to inhibit apatite formation. On the other hand, another NaOH reagent with a nominal purity of just 97 % did not exhibit any such inhibition, since it contained almost no Ca contamination. This indicates that NaOH reagent must be carefully selected for obtaining reliable bone-bonding implants of Ti metal by the NaOH and heat treatments.

Osteoconduction of porous Ti metal enhanced by acid and heat treatments.

Bone ingrowth into porous Ti metal is important for stable fixation of Ti metal implants to surrounding bone. However, without surface treatment this is limited to only a thin region of the outer surface of the Ti metal. In the present study, a porous Ti metal with a porosity of ~60 % and interpore connections of 70-200 micrometers in diameter was investigated in terms of its chemical and heat treatments, by implanting it into rabbit femur for periods varying from 3 to 12 weeks. The porous Ti metal subjected to heat treatment at 600 °C after H2SO4/HCl mixed acid treatment showed the largest bone ingrowth in comparison with those subjected to no treatment, only acid treatment, and only heat treatment even at an early stage after implantation, and remained as such even 12 weeks after implantation. Their bone ingrowths were well interpreted in terms of apatite-forming abilities of the Ti metals in body environment. Their apatite-forming abilities did not depend upon their surface roughness nor type of crystalline phase, but upon the positive surface charge.

Formation of a bioactive calcium titanate layer on gum metal by chemical treatment.

The so-called gum metal with the composition Ti-36Nb-2Ta-3Zr-0.3O is free from cytotoxic elements and exhibits a low elastic modulus as well as high mechanical strength. In the present study, it was shown that this alloy exhibited a high capacity for apatite formation in a simulated body fluid when subjected to 1 M NaOH treatment, 100 mM CaCl(2) treatment, heat treatment at 700°C, and then hot water treatment. The high apatite formation was attributed to the CaTi(2)O(5) which was precipitated on its surface, and found to be maintained even in a humid environment over a long period. The treated surface exhibited high scratch resistance, which is likely to be useful in clinical applications. The surface treatment had little effect on the unique mechanical properties described above. These results show that gum metal subjected to the present surface treatments exhibits a high potential for bone-bonding, which will be useful in orthopedic and dental implants.

Bone-bonding properties of Ti metal subjected to acid and heat treatments.

The effects of surface treatment on the bone-bonding properties of Ti metal were examined by both mechanical detaching test and histological observation after implantation into rabbit tibiae for various periods ranging from 4 to 26 weeks. The bone-bonding ability of Ti metal, which is extremely low as it is abraded, was hardly increased by simple heat treatment at 600 °C or treatment with H(2)SO(4)/HCl mixed acid alone, but was markedly increased by the heat treatment after the acid treatment. Even Ti metal that had been previously subjected to NaOH treatment showed considerably high bone-bonding ability after acid and heat treatments. Such high bonding abilities were attributed to their high apatite-forming ability in the body environment. Their high apatite-forming abilities were attributed to a high positive surface charge, and not to the type of crystalline phase or specific roughness of their surfaces. The present study has demonstrated that acid and subsequent heat treatments are effective for conferring stable fixation properties on Ti metal implants.

Evaluation of lung perfusion by using lung perfusion SPECT and lung CT with breathing synchronization software.

BACKGROUND: Lung perfusion using 99mTc-macroaggregated albumin single-photon emission computed tomography (SPECT) and lung computed tomography (CT) is a useful modality for identifying patients with pulmonary artery embolism. However, conformity between SPECT and CT at the bottom of the lung is generally low. This study aims to investigate the progression of conformity between lung perfusion SPECT and lung CT using a breathing synchronization software. METHODS: Among 95 consecutive patients who underwent lung perfusion SPECT and lung CT within 14 days because of suspected pulmonary embolism between June 2019 and August 2020 in department of cardiovascular medicine, we identified 28 patients (73 ± 10 years) who had normal pulmonary artery on contrast lung CT. We compared lung volumes calculated using lung perfusion SPECT and lung CT as gold standard. Visual conformity between lung SPECT and lung CT was scored 0-4 (0: 0-25%, 1: 25-50%, 2: 50-75%, 3: 75-90%, 4: > 90%) by two specialists in nuclear medicine and assessed. RESULTS: The lung volume calculated from lung CT was 3749 ± 788 ml. The lung volume calculated from lung perfusion SPECT without using the breathing synchronization software was 3091 ± 610 ml. There was a significant difference between the lung volume calculated from CT and SPECT without using the breathing synchronization software (P < 0.01). The lung volume calculated from lung perfusion SPECT using the breathing synchronization software was 3435 ± 686 ml, and there was no significant difference between the lung volume calculated from CT and SPECT using the breathing synchronization software. The visual score improved with the use of breathing synchronization software (without software; 1.9 ± 0.6 vs. with software; 3.4 ± 0.7, P < 0.001). CONCLUSION: This study demonstrated that the breathing synchronization software could improve conformity between lung perfusion SPECT and lung CT.

A novel synthetic material for spinal fusion: a prospective clinical trial of porous bioactive titanium metal for lumbar interbody fusion.

The objective of this study was to establish the efficacy and safety of porous bioactive titanium metal for use in a spinal fusion device, based on a prospective human clinical trial. A high-strength spinal interbody fusion device was manufactured from porous titanium metal. A bioactive surface was produced by simple chemical and thermal treatment. Five patients with unstable lumbar spine disease were treated surgically using this device in a clinical trial approved by our Ethics Review Committee and the University Hospital Medical Information Network. Clinical and radiological results were reported at the minimum follow-up period of 1 year. The optimal mechanical strength and interconnected structure of the porous titanium metal were adjusted for the device. The whole surface of porous titanium metal was treated uniformly and its bioactive ability was confirmed before clinical use. Successful bony union was achieved in all cases within 6 months without the need for autologous iliac crest bone grafting. Two specific findings including an anchoring effect and gap filling were evident radiologically. All clinical parameters improved significantly after the operation and no adverse effects were encountered during the follow-up period. Although a larger and longer-term follow-up clinical study is mandatory to reach any firm conclusions, the study results show that this porous bioactive titanium metal is promising material for a spinal fusion device.

Nanostructured positively charged bioactive TiO2 layer formed on Ti metal by NaOH, acid and heat treatments.

Nanometer-scale roughness was generated on the surface of titanium (Ti) metal by NaOH treatment and remained after subsequent acid treatment with HCl, HNO(3) or H(2)SO(4) solution, as long as the acid concentration was not high. It also remained after heat treatment. Sodium hydrogen titanate produced by NaOH treatment was transformed into hydrogen titanate after subsequent acid treatment as long as the acid concentration was not high. The hydrogen titanate was then transformed into titanium oxide (TiO(2)) of anatase and rutile by heat treatment. Treated Ti metals exhibited high apatite-forming abilities in a simulated body fluid especially when the acid concentration was greater than 10 mM, irrespective of the type of acid solutions used. This high apatite-forming ability was maintained in humid environments for long periods. The high apatite-forming ability was attributed to the positive surface charge that formed on the TiO(2) layer and not to the surface roughness or a specific crystalline phase. This positively charged TiO(2) induced apatite formation by first selectively adsorbing negatively charged phosphate ions followed by positively charged calcium ions. Apatite formation is expected on the surfaces of such treated Ti metals after short periods, even in living systems. The bonding of metal to living bone is also expected to take place through this apatite layer.

Effect of heat treatments on apatite-forming ability of NaOH- and HCl-treated titanium metal.

Titanium (Ti) metal was soaked in HCl solution after NaOH treatment and then subjected to heat treatments at different temperatures. Their apatite-forming abilities in a simulated body fluid (SBF) were discussed in terms of their surface structures and properties. The nanometer scale roughness formed on Ti metal after NaOH treatment remained after the HCl treatment and a subsequent heat treatment below 700°C. Hydrogen titanate was formed on Ti metal from an HCl treatment after NaOH treatment, and this was converted into titanium oxide of anatase and rutile phases by a subsequent heat treatment above 500°C. The scratch resistance of the surface layer increased with the formation of the titanium oxide after a heat treatment up to 700°C, and then decreased with increasing temperature. The Ti metal with a titanium oxide layer formed on its surface showed a high apatite-forming ability in SBF when the heat treatment temperature was in the range 500-700°C. The high apatite-forming ability was attributed to the positive surface charge in an SBF. These positive surface charges were ascribed to the presence of chloride ions, which were adsorbed on the surfaces and dissociated in the SBF to give an acid environment.

Stand-Alone Anterior Cervical Discectomy and Fusion Using an Additive Manufactured Individualized Bioactive Porous Titanium Implant without Bone Graft: Results of a Prospective Clinical Trial.

The purpose of this study was to introduce our patient-specific bioactive porous titanium implant manufactured using selective laser melting (SLM) and to establish the efficacy and safety of the implant for stand-alone anterior cervical discectomy and fusion (ACDF) based on a prospective clinical trial. We designed a customized ACDF implant using patient-specific data and manufactured the implant using SLM. We produced a bioactive surface through a specific chemical and thermal treatment. Using this implant, we surgically treated four patients with cervical degenerative disc disease and evaluated the clinical and radiological results. We achieved successful bony union in all but one patient without autologous bone grafting within 1 year. We observed no implant subsidence during the follow-up period, and all clinical parameters improved significantly after surgery, with no reported implant-related adverse effects. Our customized bioactive porous titanium implant is a safe and promising implant for stand-alone ACDF.

Osteoconductivity of bioactive Ti-6Al-4V implants with lattice-shaped interconnected large pores fabricated by electron beam melting.

Additive manufacturing has facilitated the fabrication of orthopedic metal implants with interconnected pores. Recent reports have indicated that a pore size of 600 µm is beneficial for material-induced osteogenesis. However, the complete removal of the metal powder from such small pores of implants is extremely difficult especially in electron beam melting (EBM). We therefore developed a new type of Ti-6Al-4V implant with lattice-shaped interconnected pores measuring 880-1400 µm, which allowed for the easy removal of metal powder. This implant was fabricated by EBM and treated with NaOH, CaCl2, heat, and water (ACaHW treatment) to render the metal surface bioactivity. In the present study, the mechanical and chemical property of the implants and the biocompatibility were evaluated. The SEM and micro-CT images demonstrated the 3D interconnectivity of the porous structures. The average porosity of the porous titanium implant was 57.5%. The implant showed maximum compressive load of 78.9 MPa and Young's modulus of 3.57 GPa which matches that of human cortical bone. ACaHW treatment of the porous Ti-6Al-4V implants induced apatite formation in simulated body fluid in vitro. The ACaHW-treated porous implants harvested from rabbit femoral bone showed direct bonding of bone to the metal surface without interposition of fibrous tissue. The porous ACaHW-treated implant had a higher affinity to the bone than the untreated one. The mechanical strength of implant fixation assessed using the push-out test was significantly higher in the ACaHW-treated implant than in untreated one. FE-SEM analysis and EDX mapping after push-out test of solid implants showed a lot of bone tissue patches on the surface of the ACaHW-treated implant. These results suggest that the new ACaHW-treated Ti-6Al-4V implant with lattice-shaped interconnected pores is a superior alternative to conventional materials for medical application.

Apatite-forming ability of Ti-15Zr-4Nb-4Ta alloy induced by calcium solution treatment.

Ti-15Zr-4Nb-4Ta alloy free from cytotoxic elements shows high mechanical strength and high corrosion resistance. However, simple NaOH and heat treatments cannot induce its ability to form apatite in the body environment. In the present study, this alloy was found to exhibit high apatite-forming ability when it was treated with NaOH and CaCl(2) solutions, and then subjected to heat and hot water treatments to form calcium titanate, rutile, and anatase on its surface. Its high apatite-forming ability was maintained even in 95% relative humidity at 80 degrees C after 1 week. The surface layer of the treated alloy had scratch resistance high enough for handling hard surgical devices. Thus, the treated alloy is believed to be useful for orthopedic and dental implants.

Prediction of osteoconductive activity of modified potassium fluorrichterite glass-ceramics by immersion in simulated body fluid.

Potassium fluorrichterite (KNaCaMg(5)Si(8)O(22)F(2)) glass-ceramics were modified by either increasing the concentration of calcium (GC5) or by the addition of P(2)O(5) (GP2). The stoichiometric composition (GST), GC5 and GP2 were soaked in simulated body fluid (SBF) along with 45S5-type bioglass as a control. After immersion, surface analyses were performed using thin-film X-ray diffraction (TF-XRD), scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS) and Fourier-transform infrared (reflection) spectroscopy (FT-IR). All compositions showed the formation of a calcium phosphate rich surface layer in SBF; GST, GP2 and the bioglass control within 7 days of immersion and GC5 after 14 days. It was concluded that all compositions were likely to be osteoconductive in vivo, with GP2 providing the best performance in terms of the combination of rapid formation of the surface layer and superior mechanical properties. This glass-ceramic system has potential as a load bearing bioceramic for fabrication of medical devices intended for skeletal tissue repair.

Hydrocolloid dressing improves wound healing by increasing M2 macrophage polarization in mice with diabetes.

Impaired wound healing is one of the most common complications of diabetes, and is known to be caused by multiple complicated factors. For instance, impaired angiogenesis, neuropathy, and hyperglycemia all function to delay subsequent wound closure. Alternatively, moist wound healing, which provides an appropriate environment for wounds, was reported to permit rapid healing by managing wound exudate. Accordingly, wound dressing materials that facilitate moist healing have been developed. The present study sought to clarify the effects of wound dressing material for moist healing of diabetic wounds, in terms of the dynamics of angiogenic factors and macrophages, using a mouse model of naturally occurring diabetes. Wounds with full-thickness skin defects were inflicted on the backs of mice and covered with dressing materials of hydrogel or gauze (control), which were retained for 3, 5, 7, 10, or 14 days following wound generation. During this time, the localization of neutrophils, fibroblasts and macrophages as well as the expression of vascular endothelial growth factor (VEGF) in the wounds and surrounding areas was observed each day. Healing clearly occurred in the hydrogel group with an increase in neutrophils and the angiogenic factor, VEGF. Moreover, the use of hydrogel resulted in a rapid rise in M1 macrophages, which appeared in the early stage of the injury, as well as rapid subsequent appearance of M2 macrophages. Thus, herein, we demonstrate that the formation of a moist environment via wound dressing material effectively improves diabetic wound healing.