Temperature-responsive bioactive glass/polymer hybrids allow for tailoring of ion release.

Intelligent biomaterials react to their surrounding conditions, and hybrid materials are acknowledged for their remarkable customizability, achieved through the meticulous control of nanoscale interactions between organic and inorganic phases. Bioactive glasses (BG) are used clinically to regenerate bone due to their degradability, ion release, and capacity to stimulate the formation of new body tissue. In our study, we developed a core-shell hybrid system using sol-gel derived BG nano particles as the core and poly (N-isopropyl acrylamide) (PNIPAM) as the shell. This approach aims to combine the therapeutic ion release of BG with the temperature-responsive properties of PNIPAM. Our size analysis by dynamic light scattering at varying temperatures shows the formation of BG aggregates driven by the coil-to-globule transition of PNIPAM on the BG surface. This transition also affected the ion release from the core-shell system through an increase in ion transport through the porous hybrid network. Our study therefore illustrates the ability to adjust the dissolution properties of the core-shell system via surrounding temperature and, thus, control the release of Ca ions from the BG.

Dispersion, ionic bonding and vibrational shifts in phospho-aluminosilicate glasses.

Understanding the relationships between structure and properties of aluminosilicate glasses is of interest in magmatic studies as well as for glass applications as mechanical or optical components. Glass properties may be tailored by the incorporation of additional elements, and here we studied the effect of phosphate incorporation on refractive index and the degree of ionic bonding in aluminosilicate glasses. The studied glasses in the system SiO2-Al2O3-Na2O-P2O5 had a metaluminous composition (Al:Na = 1) with the content of SiO2 ranging from 50 to 70 mol% and of P2O5 from 0 to 7.5 mol%. Refractive index was measured at four wavelengths from visible to near-infrared and found to decrease both with increasing P2O5 content (at the expense of NaAlO2) and with increasing SiO2 content (by substitution of SiO4 for AlO4 groups). This trend correlated with a decrease in density while, additionally, the formation of Al-O-P bonds with an SiO2-like structure may account for this change. The degree of ionic bonding, assessed via optical basicity and oxygen polarisability, decreased with increasing P2O5 and SiO2 content. Despite the complexity of the studied glasses, oxygen polarisability and optical basicity were found to follow Duffy's empirical equation for simple oxide glasses. In the high frequency infrared and Raman spectra, band shifts were observed with increasing P2O5 and SiO2 content. They indicated changing average bond strength of the glass network and showed a linear correlation with optical basicity.

High chloride content calcium silicate glasses.

Chloride is known to volatilize from silicate glass melts and until now, only a limited number of studies on oxychloride silicate glasses have been reported. In this paper we have synthesized silicate glasses that retain large amounts of CaCl2. The CaCl2 has been added to the calcium metasilicate composition (CaO·SiO2). Glasses were produced via a melt quench route and an average of 70% of the chloride was retained after melting. Up to 31.6 mol% CaCl2 has been successfully incorporated into these silicate glasses without the occurrence of crystallization. 29Si MAS-NMR spectra showed the silicon being present mainly as a Q2 silicate species. This suggests that chloride formed Cl-Ca(n) species, rather than Si-Cl bonds. Upon increasing the CaCl2 content, the Tg reduced markedly from 782 °C to 370 °C. Glass density and glass crystallization temperature decreased linearly with an increase in the CaCl2 content. However, both linear regressions revealed a breakpoint at a CaCl2 content just below 20 mol%. This might be attributed to a significant change in the structure and is also correlated with the nature of the crystallizing phases formed upon heat treatment. The glasses with less than 19.2 mol% CaCl2 crystallized to wollastonite, whilst the compositions with CaCl2 content equal to or greater than 19.2 mol% are thought to crystallize to CaCl2. In practice, the crystallization of CaCl2 could not occur until the crystallization temperature fell below the melting point of CaCl2. The implications of the results along with the high chloride retention are discussed.

Well-Defined SiO2@P(EtOx-stat-EI) Core-Shell Hybrid Nanoparticles via Sol-Gel Processes.

Positively charged nanoparticles (NPs) are very interesting for biomedical and pharmaceutical applications, such as nonviral gene delivery. Here, the synthesis of SiO2 nanoparticles with a covalently grafted poly(2-ethyl-2-oxazoline) (PEtOx) shell (SiO2@PEtOx) is presented. PEtOx with a degree of polymerization of 20 and 38 is synthesized via microwave supported cationic ring-opening polymerization and subsequently end-functionalized with a triethoxysilyl linker for subsequent grafting to silica particles with hydrodynamic radii of 7, 31, and 152 nm. The resulting SiO2@PEtOx particles are characterized by using dynamic light scattering (DLS), transmission electron microscopy (TEM, cryoTEM), and scanning electron microscopy (SEM) to determine changes in particle size. Thermal gravimetrical analysis is used to quantify the amount of polymer on the silica surface. Subsequent in situ transformation of SiO2@PEtOx particles into SiO2@P(EtOx-stat-EI) (poly(2-ethyl-2-oxazoline-stat-ethylene imine) grafted silica particles) under acidic conditions inverts the surface charge from negative to positive according to ζ-potential measurements. The P(EtOx-stat-EI) shell could be used for the deposition of Au NP afterward.

Bioactive glasses­structure and properties.

Bioactive glasses were the first synthetic materials to show bonding to bone, and they are successfully used for bone regeneration. They can degrade in the body at a rate matching that of bone formation, and through a combination of apatite crystallization on their surface and ion release they stimulate bone cell proliferation, which results in the formation of new bone. Despite their excellent properties and although they have been in clinical use for nearly thirty years, their current range of clinical applications is still small. Latest research focuses on developing new compositions to address clinical needs, including glasses for treating osteoporosis, with antibacterial properties, or for the sintering of scaffolds with improved mechanical stability. This Review discusses how the glass structure controls the properties, and shows how a structure-based design may pave the way towards new bioactive glass implants for bone regeneration.

Antimicrobial Biomaterials Based on Physical and Physicochemical Action.

Developing effective antimicrobial biomaterials is a relevant and fast-growing field in advanced healthcare materials. Several well-known (e.g., traditional antibiotics, silver, copper etc.) and newer (e.g., nanostructured, chemical, biomimetic etc.) approaches have been researched and developed in recent years and valuable knowledge has been gained. However, biomaterials associated infections (BAIs) remain a largely unsolved problem and breakthroughs in this area are sparse. Hence, novel high risk and potential high gain approaches are needed to address the important challenge of BAIs. Antibiotic free antimicrobial biomaterials that are largely based on physical action are promising, since they reduce the risk of antibiotic resistance and tolerance. Here, selected examples are reviewed such antimicrobial biomaterials, namely switchable, protein-based, carbon-based and bioactive glass, considering microbiological aspects of BAIs. The review shows that antimicrobial biomaterials mainly based on physical action are powerful tools to control microbial growth at biomaterials interfaces. These biomaterials have major clinical and application potential for future antimicrobial healthcare materials without promoting microbial tolerance. It also shows that the antimicrobial action of these materials is based on different complex processes and mechanisms, often on the nanoscale. The review concludes with an outlook and highlights current important research questions in antimicrobial biomaterials.

Bioactive glass-ceramics containing fluorapatite, xonotlite, cuspidine and wollastonite form apatite faster than their corresponding glasses.

Crystallisation of bioactive glasses has been claimed to negatively affect the ion release from bioactive glasses. Here, we compare ion release and mineralisation in Tris-HCl buffer solution for a series of glass-ceramics and their parent glasses in the system SiO2-CaO-P2O5-CaF2. Time-resolved X-ray diffraction analysis of glass-ceramic degradation, including quantification of crystal fractions by full pattern refinement, show that the glass-ceramics precipitated apatite faster than the corresponding glasses, in agreement with faster ion release from the glass-ceramics. Imaging by transmission electron microscopy and X-ray nano-computed tomography suggest that this accelerated degradation may be caused by the presence of nano-sized channels along the internal crystal/glassy matrix interfaces. In addition, the presence of crystalline fluorapatite in the glass-ceramics facilitated apatite nucleation and crystallisation during immersion. These results suggest that the popular view of bioactive glass crystallisation being a disadvantage for degradation, apatite formation and, subsequently, bioactivity may depend on the actual system study and, thus, has to be reconsidered.

Lithiated porous silicon nanowires stimulate periodontal regeneration.

Periodontal disease is a significant burden for oral health, causing progressive and irreversible damage to the support structure of the tooth. This complex structure, the periodontium, is composed of interconnected soft and mineralised tissues, posing a challenge for regenerative approaches. Materials combining silicon and lithium are widely studied in periodontal regeneration, as they stimulate bone repair via silicic acid release while providing regenerative stimuli through lithium activation of the Wnt/ß-catenin pathway. Yet, existing materials for combined lithium and silicon release have limited control over ion release amounts and kinetics. Porous silicon can provide controlled silicic acid release, inducing osteogenesis to support bone regeneration. Prelithiation, a strategy developed for battery technology, can introduce large, controllable amounts of lithium within porous silicon, but yields a highly reactive material, unsuitable for biomedicine. This work debuts a strategy to lithiate porous silicon nanowires (LipSiNs) which generates a biocompatible and bioresorbable material. LipSiNs incorporate lithium to between 1% and 40% of silicon content, releasing lithium and silicic acid in a tailorable fashion from days to weeks. LipSiNs combine osteogenic, cementogenic and Wnt/ß-catenin stimuli to regenerate bone, cementum and periodontal ligament fibres in a murine periodontal defect.

The structural role and coordination environment of cobalt in 45P2O5-CaO-Na2O phosphate glasses: thermal properties and Raman, UV-vis-NIR, and EPR spectroscopy.

Cobalt-containing materials are of interest for a wide range of applications, from biomaterials to solid-state lasers in optics. For instance, Co2+ is known to trigger the formation of new blood vessels, i.e. angiogenesis. Here, the use of phosphate glasses as a vehicle for local release of Co2+ ions is an attractive strategy to overcome the vascularisation limitation in tissue engineering. This study aimed to establish structure-property correlations as a function of the coordination environment of cobalt in 45P2O5-(30 - x)CaO-25Na2O-xCoO (x: 0.01 to 10 mol%) glasses. Constant polymerization and O/P ratio, resulting ultimately in constant basicity, were shown by ICP-OES and Raman spectroscopy. The latter, combined with EPR analysis, indicated that Co2+ was the predominant oxidation state and the presence of Co3+ can be excluded. UV-vis-NIR absorption spectra showed that the ratio between Co2+ in four- and six-fold coordination remained constant throughout the glass series. Their thermal properties measured by DSC and heating microscopy did not change much in the substitution range studied here. The steady trend in Tg values suggests a compensation between two opposite effects caused by the presence of four and six-fold coordinated Co2+, both being present at a constant ratio throughout the glasses. Accordingly, the higher field strength of Co2+ compared to that of Ca2+ is expected to strengthen the glass network. In contrast, four-fold coordinated cobalt is expected to weaken the network by connecting fewer fragments of the phosphate glass network than six-fold coordinated cobalt. These results indicate that the structural properties of the glasses with constant basicity are influenced by the coordination number of Co2+.

Tissue-specific calibration of extracellular matrix material properties by transforming growth factor-ß and Runx2 in bone is required for hearing.

Physical cues, such as extracellular matrix stiffness, direct cell differentiation and support tissue-specific function. Perturbation of these cues underlies diverse pathologies, including osteoarthritis, cardiovascular disease and cancer. However, the molecular mechanisms that establish tissue-specific material properties and link them to healthy tissue function are unknown. We show that Runx2, a key lineage-specific transcription factor, regulates the material properties of bone matrix through the same transforming growth factor-ß (TGFß)-responsive pathway that controls osteoblast differentiation. Deregulated TGFß or Runx2 function compromises the distinctly hard cochlear bone matrix and causes hearing loss, as seen in human cleidocranial dysplasia. In Runx2+/⁻ mice, inhibition of TGFß signalling rescues both the material properties of the defective matrix, and hearing. This study elucidates the unknown cause of hearing loss in cleidocranial dysplasia, and demonstrates that a molecular pathway controlling cell differentiation also defines material properties of extracellular matrix. Furthermore, our results suggest that the careful regulation of these properties is essential for healthy tissue function.

Influence of Phosphate on Network Connectivity and Glass Transition in Highly Polymerized Aluminosilicate Glasses.

Melt-derived metaluminous (Al/Na = 1) aluminosilicate glasses in the system SiO2-Al2O3-Na2O-P2O5 were prepared with P2O5 and SiO2 contents varying from 0 to 7.5 and 50 to 70 mol %, respectively. The glass structure was investigated by X-ray absorption near edge structure, far- and medium-infrared, and polarized Raman spectroscopic techniques. The results indicate the incorporation of phosphate into the aluminosilicate network not only as partially depolymerized groups but also as fully polymerized groups charge-balanced by aluminate units in Al-O-P bonds. A new analysis method based on polarized Raman spectra in the bending frequency range indicates a preference of phosphate to reorganize the smallest ring structures. Changes in the glass transition temperature with the increase in phosphate content were found to be consistent with the depolymerization of the network structure shown by spectroscopy. By contrast, increasing the silica content by substituting SiO4 for AlO4 tetrahedra, while keeping the phosphate content constant, was found to have a negligible effect on network polymerization. Still, the glass transition temperature decreased and correlated with a far-infrared sodium band shift to higher frequency. This was interpreted as local changes in bond strength caused by complex interactions between the different network formers and sodium ions.

A modified glass ionomer cement to mediate dentine repair.

OBJECTIVES: Glass ionomer cements (GIC) can be used to protect dentine following caries removal. However, GIC have little biological activity on biological repair processes, which means that neo-dentine formation remains reliant on limited endogenous regenerative processes. Wnt/ß-catenin signalling is known to play a central role in stimulating tertiary dentine formation following tooth damage and can be stimulated by a range of glycogen synthase kinase (GSK3) antagonists, including lithium ions. METHODS: Here, we created lithium-containing bioactive glass (BG) by substituting lithium for sodium ions in 45S5 BG. We then replaced between 10 and 40% of the powder phase of a commercial GIC with the lithium-substituted BG to create a range of formulations of 'LithGlassGIC'. In vitro physical properties of the resulting glasses were characterised and their ability to stimulate reactionary dentine formation in mouse molars in vivo was tested. RESULTS: Lithium release from LithGlassGIC increased with increasing lithium content in the cement. In common with unmodified commercial GIC, all formations of LithGlassGIC showed in vitro toxicity when measured using an indirect cell culture assay based on ISO10993:5, precluding direct pulp contact. However, in a murine non-exposed pulp model of tooth damage, LithGlassGIC quickly released lithium ions, which could be transiently detected in the saliva and blood. LithGlassGIC also enhanced the formation of tertiary dentine, resulting in a thickening of the dentine at the damage site that restored lost dentine volume. Dentine regeneration was likely mediated by upregulation of Wnt/ß-catenin activity, as LithGlassGIC placed in TCF/Lef:H2B-GFP reporter mice showed enhanced GFP activity. SIGNIFICANCE: We conclude that LithGlassGIC acts as a biological restorative material that promotes tertiary dentine formation and restores tooth structure.

Unravelling the dissolution mechanism of polyphosphate glasses by 31P NMR spectroscopy: ligand competition and reactivity of intermediate complexes.

Phosphate glass dissolution can be tailored via compositional and subsequent structural changes, which is of interest for biomedical applications such as therapeutic ion delivery. Here, solid-state 31P nuclear magnetic resonance characterisation of 45P2O5-xCaO - (55 -x)Na2O glasses was correlated with dissolution studies using time-dependent liquid 31P NMR spectroscopy and quantitative chemical analysis. Glasses dissolved congruently in aqueous media, and the first dissolution stage was the hydration of phosphate chains. In deionised water and Tris buffer (pH0 7.4 or 7.9), trimetaphosphate rings and orthophosphates were the predominant species in solution, indicating relatively fast degradation. By contrast, long phosphate chains were identified in EDTA (pH0 10.0). Besides pH differences, coordination of phosphate species by metal cations appears to play a catalytic role in the hydrolysis mechanism via turning phosphorus atoms into suitable electrophiles for the subsequent nucleophilic attack by water. Hydrolysis rates were proportional to phosphate complex stability, with stronger complexes for chains than for rings. A competition between solvent and phosphate species for the metal ion occurred in the order EDTA > Tris > deionised water.

Nano-imaging confirms improved apatite precipitation for high phosphate/silicate ratio bioactive glasses.

Bioactive glasses convert to a biomimetic apatite when in contact with physiological solutions; however, the number and type of phases precipitating depends on glass composition and reactivity. This process is typically followed by X-ray diffraction and infrared spectroscopy. Here, we visualise surface mineralisation in a series of sodium-free bioactive glasses, using transmission electron microscopy (TEM) with energy-dispersive X-ray spectroscopy (EDXS) and X-ray nano-computed tomography (nano-CT). In the glasses, the phosphate content was increased while adding stoichiometric amounts of calcium to maintain phosphate in an orthophosphate environment in the glass. Calcium fluoride was added to keep the melting temperature low. TEM brought to light the presence of phosphate clustering and nearly crystalline calcium fluoride environments in the glasses. A combination of analytical methods, including solid-state NMR, shows how with increasing phosphate content in the glass, precipitation of calcium fluoride during immersion is superseded by fluorapatite precipitation. Nano-CT gives insight into bioactive glass particle morphology after immersion, while TEM illustrates how compositional changes in the glass affect microstructure at a sub-micron to nanometre-level.

A review of in vitro cell culture testing methods for bioactive glasses and other biomaterials for hard tissue regeneration.

Bioactive glasses are used to regenerate bone by a mechanism which involves surface degradation, the release of ions such as calcium, soluble silica and phosphate and the precipitation of a biomimetic apatite surface layer on the glass. One major area of bioactive glass research is the incorporation of therapeutically active ions to broaden the application range of these materials. When developing such new compositions, in vitro cell culture studies are a key part of their characterisation. However, parameters of cell culture studies vary widely, and depending on the intended use of bioactive glass compositions, different layouts, cell types and assays need to be used. The aim of this publication is to provide materials scientists, particularly those new to cell culture studies, with a tool for selecting the most appropriate assays to give insight into the properties of interest.

Mg or Zn for Ca substitution improves the sintering of bioglass 45S5.

Bioglass 45S5 is well-known for its bioactivity, but it possesses poor sintering behaviour owing to viscous flow being inhibited by the crystallisation of sodium calcium silicate phases. Mg or Zn were partially (0, 25, 50, 75%) or fully (100%) substituted for Ca on a molar base, and thermal properties (differential scanning calorimetry, dilatometry) and sintering (heating microscopy, SEM and X-ray diffraction) were investigated. Here we show that sintering can be improved significantly by partial or complete substitution of Mg or Zn for Ca, owing to a pronounced decrease in crystallisation tendency. Glass transition temperature and dilatometric softening point went through minima for mixed compositions, with random mixing of Mg/Ca or Zn/Ca ions in the glass structure and the resulting effect on configurational entropy being a likely explanation. As the onset of crystallisation did not vary much with substitution, substituted glasses possessed a wider temperature range for sintering, resulting in up to 57% and 27% sample height reduction for Mg and Zn substituted glasses, respectively, compared to only 3% height reduction for Bioglass 45S5. Taken together, these results suggest that using a combination of modifiers, particularly alkaline earths or zinc, may be a promising approach for improving the sintering of Bioglass 45S5.

Structural Role of Phosphate in Metaluminous Sodium Aluminosilicate Glasses As Studied by Solid State NMR Spectroscopy.

In this contribution we present a detailed study of the effect of the addition of small to intermediate amounts of P2O5 (up to 7.5 mol %) on the network organization of metaluminous sodium aluminosilicate glasses employing a range of advanced solid state NMR methodologies. The combined results from MAS, MQMAS (multiple quantum MAS), or MAT (magic angle turning) NMR spectroscopy and a variety of dipolar based NMR experiments-27Al{31P}-, 27Al{29Si}-, 29Si{31P}-, and 31P{29Si}-REDOR (rotational echo double resonance) NMR spectroscopy as well as 31P{27Al}- and 29Si{27Al}-REAPDOR (rotational echo adiabatic passage double resonance) NMR-allow for a detailed analysis of the network organization adopted by these glasses. Phosphate is found as QP2, QP3, and QP4 (with the superscript denoting the number of bridging oxygens), the QP4 units can be safely identified with the help of 31P MAT NMR experiments. Al exclusively adopts a 4-fold coordination. The withdrawal of a fraction of the sodium cations from AlO4 units that is needed for charge compensation of the QP2 units necessitates an alternative charge compensation scheme for these AlO4 units via formation of QP4 units or oxygen triclusters. The dipolar NMR experiments suggest a strong preference of P for Al with an average value of ca. 2.4 P-O-Al connections per phosphate tetrahedron. P is thus mainly integrated into the network via P-O-Al bonding, the formation of Si-O-P bonding plays only a minor role.

A comparison of lithium-substituted phosphate and borate bioactive glasses for mineralised tissue repair.

OBJECTIVES: Wnt/ß-catenin signalling plays important roles in regeneration, particularly in hard tissues such as bone and teeth, and can be regulated by small molecule antagonists of glycogen synthase kinase 3 (GSK3); however, small molecules can be difficult to deliver clinically. Lithium (Li) is also a GSK3 antagonist and can be incorporated into bioactive glasses (BG), which can be used clinically in dental and bone repair applications and tuned to quickly release their constituent ions. METHODS: Here, we created phosphate (P)- and borate (B)-based BG that also contained Li (LiPBG and LiBBG) and examined their ion release kinetics and the toxicity of their dissolution ions on mouse 17IA4 dental pulp cells. RESULTS: We found that although LiPBG and LiBBG can both quickly release Li at concentrations known to regulate Wnt/ß-catenin signalling, the P and B ions they concomitantly release are highly toxic to cells. Only when relatively low concentrations of LiPBG and LiBBG were placed in cell culture medium were their dissolution products non-toxic. However, at these concentrations, LiPBG and LiBBG's ability to regulate Wnt/ß-catenin signalling was limited. SIGNIFICANCE: These data suggest that identifying a BG composition that can both quickly deliver high concentrations of Li and is non-toxic remains a challenge.

Effect of sterilization by gamma radiation on nano-mechanical properties of teeth.

OBJECTIVES: Extracted teeth used in dental research need to be considered infective and hence be sterilized without the materials' properties being altered. This study examined the effect of gamma radiation on the nano-mechanical properties of dentin and enamel of extracted human third molars. METHODS: Whole teeth were sterilized using gamma radiation doses of 7 kGy and 35 kGy, respectively; teeth of the control group were not treated with gamma radiation. Crowns were sectioned occlusally and polished. Elastic modulus and hardness were tested using atomic force microscopy with nano-indentations under wet conditions. RESULTS: The authors found no significant dose-response relationship in elastic modulus or hardness in either dentin or enamel. SIGNIFICANCE: Nano-indentation is a common technique for the determination of local mechanical properties in biological hard tissues. Gamma radiation is an efficient way to sterilize extracted teeth while alteration of dentin and enamel mechanical properties are minimized.

In-vitro apatite formation capacity of a bioactive glass - containing toothpaste.

OBJECTIVES: The in-vitro dissolution of bioactive glass-based toothpastes and their capacity to form apatite-like phases in buffer solutions have been investigated. MATERIALS AND METHODS: The commercial toothpaste samples were tested on immersion in artificial saliva, Earle's salt solution and Tris buffer for duration from 10min to four days. The powder samples collected at the end of the immersion were studied using solid-state 31P and 19F nuclear magnetic resonance spectroscopy (NMR), X-ray powder diffraction and Fourier transform infrared (FTIR) spectroscopy. The fluoride concentration in the solution remained after the immersion was measured. RESULTS: In artificial saliva and in presence of sodium monofluorophosphate (MFP), the bioactive glass and bioactive glass-based toothpastes formed fluoridated apatite-like phases in under 10min. A small amount of apatite-like phase was detected by 31P NMR in the toothpaste with MFP but no bioactive glass. The toothpaste with bioactive glass but no fluoride formed an apatite-like phase as rapidly as the paste containing bioactive glass and fluoride. By contrast, apatite-like phase formation was much slower in Earle's salt solution than artificial saliva and slower than Tris buffer. CONCLUSIONS: The results of this lab-based study showed that the toothpaste with MFP and bioactive glass formed a fluoridated apatite in artificial saliva and in Tris buffer, as did the mixture of bioactive glass and MFP. CLINICAL SIGNIFICANCE: The presence of fluoride in bioactive glass-containing toothpastes can potentially lead to the formation of a fluoridated apatite, which may result in improved clinical effectiveness and durability. However, this should be further tested intra-orally.