Doxorubicin-loaded polyphosphate glass microspheres for transarterial chemoembolization.

The standard of care for intermediate stage hepatocellular carcinoma is transarterial chemoembolization (TACE). Drug-eluting bead TACE (DEB-TACE) has emerged as a leading form of TACE, as it uses highly calibrated microspheres to deliver consistent embolization and controlled drug release to the tumor microenvironment. We report here on doxorubicin (DOX)-loaded polyphosphate glass microspheres (PGM) as a novel resorbable, radiopaque, preloaded DEB-TACE platform. Coacervate composed of polyphosphate chains complexed with Ba2+ , Ca2+ , and Cu2+ can be loaded with DOX prior to PGM synthesis, with PGM production achieved using a water-in-oil emulsion technique at room temperature yielding highly spherical particles in clinically relevant size fractions. In vitro, DOX release was found to be linear, pH dependent, and in accordance with Type II non-Fickian transport. PGM degradation was characterized by an initial burst release of degradation products over 7 days, followed by a plateau in mass loss at approximately 75% over a period of several weeks. in vitro studies indicate that PGM degradation products, namely Cu2+ , are cytotoxic and may interact with eluted DOX to impair its pharmacological activity. With additional compositional considerations, this approach may prove promising for DEB-TACE applications.

Minocycline-loaded calcium polyphosphate glass microspheres as a potential drug-delivery agent for the treatment of periodontitis.

Background: Periodontitis is an inflammatory disease with a bacterial etiology that affects the supporting structures of the teeth and is a major cause of tooth loss. The objective of this study was to investigate the drug loading and in vitro release of minocycline from novel calcium polyphosphate microspheres intended for use in treating periodontitis. Methods: Calcium polyphosphate coacervate, produced by a precipitation reaction of calcium chloride and sodium polyphosphate solutions, was loaded with minocycline and subsequently used to produce microspheres by an emulsion/solvent extraction technique. Microspheres classified by size were subjected to a 7-day elution in a Tris-buffer solution under dynamic conditions. The physicochemical characteristics of the drug-loaded microspheres were investigated using scanning electron microscopy, particle size analysis, Phosphorus-31 Nuclear Magnetic Resonance spectroscopy, and Inductively Coupled Plasma Optical Emission Spectroscopy. Drug loading and release were determined using ultraviolet -visible (UV/VIS) spectrophotometry. Results: Minocycline-loaded calcium polyphosphate microspheres of varying size were successfully produced, with small and large microspheres having volume mean diameters of 22 ± 1 µm and 193 ± 5 µm, respectively. Polyphosphate chain length and calcium to phosphorus mole ratio remained stable throughout microsphere production. Drug loading was 1.64 ± 0.16, 1.35 ± 0.55, and 0.84 ± 0.14 weight% for the coacervate and large and small microspheres, respectively, corresponding to mean encapsulation efficiencies of 81.7 ± 12.2 % and 50.9 ± 3.9 % for the large and small microspheres. Sustained drug release was observed in vitro over a clinically relevant 7-day period, with small and large microspheres exhibiting similar elution profiles. Antibiotic release generally followed microsphere degradation as measured by Ca and P ion release. Conclusions: This study demonstrated successful drug loading of calcium polyphosphate microspheres with minocycline. Furthermore, in vitro sustained release of minocycline over a 7-day period was observed, suggesting potential utility of this approach for treating periodontitis.

Impact of trivalent ions on the stability and cohesion of calcium polyphosphate coacervates for embolization applications.

Polyphosphates (PPs) are of interest as temporary in situ setting embolic agents for which cohesive characteristics are vital. Trivalent ions Al3+ and Ga3+ were substituted into calcium PP up to 10 mol % for two PP chain lengths (degree of polymerization, Dp 200 and 9000) and the effect on the dissolution rate of the resulting coacervate was examined. High levels of trivalent ions were found to increase the dissolution rate, especially with aluminum (Al) where the coacervate with the greatest Al content (10 mol %) and larger Dp completely dissolved within the first few hours in tris(hydroxymethyl)aminomethane buffered saline. Conversely, small amounts of trivalent ions slowed the dissolution rate of the coacervates compared to those containing calcium only. The coacervate compositions determined to have the fastest and slowest ion release were evaluated for cohesion upon injection into a simulated blood vessel using a dual lumen needle. PPs with lower trivalent content had a higher coacervate yield overall, with 5% Ga and Dp 200 yielding the smallest proportion of coacervate particulates that could be implicated in unwanted distal embolization. However, further studies are required to evaluate the formation and duration of occlusions in vivo so that the PP composition can best be tailored to meet clinical requirements. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B:2638-2648, 2019.

A two-stage cold isostatic pressing and gelling approach for fabricating a therapeutically loaded amorphous calcium polyphosphate local delivery system.

Local delivery systems have taken on a greater clinical focus for osteomyelitis therapy owing to their ability to overcome many disadvantages of systemic delivery. This study reports for the first time the capacity to fabricate strontium- and vancomycin-doped calcium polyphosphate beads using a two-stage cold isostatic pressing and gelling approach. The fabricated beads were of uniform shape and diameter, and upon gelling exhibited reduced porosity. Of greatest significance in the subsequent in vitro study was the improvement of bead long-term structural stability upon vancomycin incorporation; a characteristic that further encourages the extended release of therapeutically relevant levels of antibiotic. Overall, this study provides support for the inclusion of a cold isostatic pressing step in the fabrication of a therapeutically loaded calcium polyphosphate bead-based local delivery system intended for osteomyelitis treatment.

Phase transformations during processing and in vitro degradation of porous calcium polyphosphates.

A 2-Step sinter/anneal treatment has been reported previously for forming porous CPP as biodegradable bone substitutes [9]. During the 2-Step annealing treatment, the heat treatment used strongly affected the rate of CPP degradation in vitro. In the present study, x-ray diffraction and (31)P solid state nuclear magnetic resonance were used to determine the phases that formed using different heat treating processes. The effect of in vitro degradation (in PBS at 37 °C, pH 7.1 or 4.5) was also studied. During CPP preparation, ß-CPP and γ-CPP were identified in powders formed from a calcium monobasic monohydrate precursor after an initial calcining treatment (10 h at 500 °C). Melting of this CPP powder (at 1100 °C), quenching and grinding formed amorphous CPP powders. Annealing powders at 585 °C (Step-1) resulted in rapid sintering to form amorphous porous CPP. Continued annealing to 650 °C resulted in crystallization to form a multi-phase structure of ß-CPP primarily plus lesser amounts of α-CPP, calcium ultra-phosphates and retained amorphous CPP. Annealing above 720 °C and up to 950 °C transformed this to ß-CPP phase. In vitro degradation of the 585 °C (Step-1 only) and 650 °C Step-2 annealed multi-phase samples occurred significantly faster than the ß-CPP samples formed by Step-2 annealing at or above 720 °C. This faster degradation was attributable to preferential degradation of thermodynamically less stable phases that formed in samples annealed at 650 °C (i.e. α-phase, ultra-phosphate and amorphous CPP). Degradation in lower pH solutions significantly increased degradation rates of the 585 and 650 °C annealed samples but had no significant effect on the ß-CPP samples.

Degradation and hemostatic properties of polyphosphate coacervates.

UNLABELLED: Sodium polyphosphate is a linear polymer formed from phosphate units linked together by sharing oxygen atoms. Addition of calcium to a solution of sodium polyphosphate results in phase separation and formation of a polyphosphate coacervate best described as a polymeric rich viscoelastic material. Polyphosphate coacervate is an interesting candidate as a biomaterial based on its ability to bind with different cations and to be loaded with drugs. Here, in vitro degradation and hemostatic properties of polyphosphate coacervates are comprehensively evaluated. We show that polyphosphate coacervates degrade and dissolve at a fast rate, losing half of their original mass in a week and transforming to mainly pyrophosphate after 4weeks. This burst dissolution phase happens earlier for the coacervate prepared from very short chain polyphosphate but overall using longer polyphosphate chains does not increase the coacervate longevity significantly. Substitution of Ca with Sr or Ba does not affect the hydrolysis of coacervates but slows down their dissolution into the media. In a whole blood clotting assay, coacervates profoundly decrease the clotting time especially when very long chain polyphosphates are used. While coacervate chain length and divalent cation type were found to significantly affect prothrombin time and thromboplastin time compared to the control, no discernible trends were observed. Platelets adhere in large numbers to coacervates, especially those containing long chain polyphosphate, but the cell morphology observed suggests that they might not to be fully activated. Overall, the long chain polyphosphate coacervate holds a great potential as a resorbable hemostatic agent. STATEMENT OF SIGNIFICANCE: Divalent cation additions to a sodium polyphosphate solution result in polyphosphate coacervates, or highly viscous gel-like materials, having great potential in bio-applications such as drug delivery and hemostasis. As these coacervates degrade in aqueous environments, we undertook a comprehensive evaluation to better understand the impact of polyphosphate chain length and divalent cation substitution on this hydrolytic response in order to better predict degradation behavior in the body. Furthermore, there is great interest in the role of polyphosphates in hemostasis following recent publications showing that platelets secrete polyphosphates upon thrombin stimulation. In this paper, we evaluate the hemostatic potential of polyphosphate coacervates as bulk constructs, demonstrating that indeed these materials hold great potential as a degradable hemostatic agent.

Developing an in situ forming polyphosphate coacervate as a new liquid embolic agent: From experimental design to pilot animal study.

A radiopaque temporary liquid embolic agent was synthesized from polyphosphate (PP) coacervates and optimized using a design of experiments approach. Variables studied were: strontium substitution (0-15 mol%), barium substitution (0-15 mol%), PP concentration and degree of polymerization of the polyphosphate (Dp). The viscosity, radiopacity and cell viability of the resulting coacervates were measured for 60 formulations and response surface modeling was used to determine the optimum coacervate that maximized radiopacity and cell viability. The optimum coacervate made from PP with a large Dp (9.5 g NaPP/100mL, 2.2 mol% Sr, 9 mol% Ba and 3.8 mol% Ca) was taken forward to a pilot animal trial. In this rabbit model, PP embolic agent successfully occluded the central auricular artery with promising biocompatibility. Further study is required to optimize the cohesiveness and clinical effectiveness of PP as an in situ setting temporary embolic agent. STATEMENT OF SIGNIFICANCE: This article describes the development of a new radiopaque temporary liquid embolic agent from the optimization using design of experiments to a pilot animal study. Embolization is a minimally invasive interventional radiology procedure used to block blood flow in a targeted blood vessel. This procedure is used to treat many conditions including: tumors, aneurysms and arteriovenous malformations. Currently, no inherent radiopaque embolic agents are available in the clinic, which would allow for direct imaging of the material during the procedure and follow up treatment.

Methotrexate-loaded glass ionomer cements for drug release in the skeleton: An examination of composition-property relationships.

Chemotherapeutic-loaded bone cement may be an effective method of drug delivery for the management of cancer-related vertebral fractures that require cement injection for pain relief. Recent advancements in the development of aluminum-free glass ionomer cements (GICs) have rendered this class of biomaterials clinically viable for such applications. To expand the therapeutic benefits of these materials, this study examined, for the first time, their drug delivery potential. Through incrementally loading the GIC with methotrexate (MTX) by up to 10-wt%, composition-property relationships were established, correlating MTX loading with working time and setting time, as well as compressive strength, drug release, and cytotoxic effect over 31 days. The most significant finding of this study was that MTX was readily released from the GIC, while maintaining cytotoxic activity. Release correlated linearly with initial loading and appeared to be diffusion mediated, delivering a total of 1-2% of the incorporated drug. MTX loading in this range exerted minimal effects to handling and strength, indicating the clinical utility of the material was not compromised by MTX loading. The MTX-GIC systems examined herein are promising materials for combined structural delivery applications.

Calcium polyphosphate as an additive to zinc-silicate glass ionomer cements.

Aluminum-free glass ionomer cements (GICs) are under development for orthopedic applications, but are limited by their insufficient handling properties. Here, the addition of calcium polyphosphate (CPP) was investigated as an additive to an experimental zinc-silicate glass ionomer cement. A 50% maximum increase in working time was observed with CPP addition, though this was not clinically significant due to the short working times of the starting zinc-silicate GIC. Surprisingly, CPP also improved the mechanical properties, especially the tensile strength which increased by ∼33% after 30 days in TRIS buffer solution upon CPP addition up to 37.5 wt%. This strengthening may have been due to the formation of ionic crosslinks between the polyphosphate chains and polyacrylic acid. Thus, CPP is a potential additive to future GIC compositions as it has been shown to improve handling and mechanical properties. In addition, CPP may stimulate new bone growth and provide the ability for drug delivery, which are desirable modifications for an orthopedic cement.

Comprehensive study of the chelation and coacervation of alkaline earth metals in the presence of sodium polyphosphate solution.

The effect of chelation of three alkaline earth metals (Ca, Sr, and Ba) by polyphosphates on the pH and viscosity of the solution is examined and correlated to the phosphate glass properties. Also, the impact of the polyphosphate average degree of polymerization (D(p)) as well as the type and amount of chelated divalent cation on the degradation rate of the chains is studied. Subsequently, the number of divalent cations required for polyphosphate chain agglomeration to form a coacervate, and the resulting composition of these coacervates, was investigated. A decrease in polyphosphate solution pH during chelation was routinely obtained, with a sudden shift in the rate of pH drop occurring around a divalent cation/phosphorus molar ratio of 0.18. Longer chains or cations with a smaller ionic radius accelerated the rate of D(p) reduction. The number of divalent cations required for coacervation depends on different variables such as the polyphosphate concentration, the D(p), and the type of divalent cation. The formed coacervate retains the D(p) of polyphosphate originally used for coacervation, and the resulting Ca/P molar ratio depends largely on the amount of calcium being used during coacervation. Overall, this article helps one to understand the coacervation of polyphosphates in order to exploit their potential as a biomaterial.

The oral health of ageing baby boomers: a comparison of adults aged 45-64 and those 65 years and older.

OBJECTIVES: To compare the oral health status of adults aged 45-64 (baby boomers) and those aged 65 and older. METHODS: An observational, cross-sectional survey of adults living independently in rural and urban settings in Nova Scotia, Canada was conducted. Using random digit dialing, calibrated interviewers completed a telephone survey, and clinicians calibrated to WHO standards conducted clinical examinations. Weighting was used to correct for sampling bias. RESULTS: 747 community dwelling adults completed both the clinical exam and the questionnaire (n=411, age 45-64; n=336, age 65 or older). Rates of edentulism were low (2.6% aged 45-64; 15.7% aged 65+; p<0.001). Untreated root caries was greater in the older dentate group (19.7 vs. 10.1%; p<0.001). Being 65 years of age or older was identified as a predictor of increased decayed, missing, filled teeth, presence of decayed and/or filled roots and presence of attachment loss≥4 mm, but was not a significant predictor of presence of untreated coronal caries. CONCLUSIONS: A falling rate of edentulism and a higher risk for root caries with increasing age may predict the need for more complex dental care as our population ages.

Oral health status of long-term care residents-a vulnerable population.

OBJECTIVE: To conduct an observational, cross-sectional survey of the oral health status of adults ≥ 45 years of age in rural and urban long-term care (LTC) facilities in Nova Scotia, Canada. METHODS: Residents capable of informed consent were recruited by LTC staff in a stratified random sample of LTC facilities. Calibrated personnel administered standard clinical and quality-of-life instruments. RESULTS: Of the 335 adults (74% female) surveyed (mean age 80.8 ± 11.6 years), only 25% reported having regular dental care. Although 76% described their oral health as good or excellent, 41% were edentulous, 41% had some mucosal abnormality, 36% reported xerostomia and 25% had perceived or self-reported untreated dental conditions. Most mandibular dentures were nonretentive (59%) and almost half were unstable (49%). Among the dentate, 51% had untreated coronal caries, 44% had untreated root caries and 67% had attachment loss of ≥ 4 mm at ≥ 1 site. Predictors of coronal decay were a debris score ≥ 2 (adjusted odds ratio [adj OR] = 2.12; p = 0.045) or a history of smoking (adj OR = 1.02 per year of smoking; p = 0.024). Predictors of root caries were participants' perceiving a need for dental treatment (adj OR = 2.56; p = 0.015) or a history of smoking (adj OR = 1.02 per year of smoking; p = 0.026). CONCLUSIONS: This epidemiologic study of the oral health of LTC residents revealed a high prevalence of untreated oral disease and low use of oral care services, highlighting the need for better access to oral care for this population.

Assessing the oral health of an ageing population: methods, challenges and predictors of survey participation.

OBJECTIVES: To examine predictors of participation and to describe the methodological considerations of conducting a two-stage population-based oral health survey. METHODS: An observational, cross-sectional survey (telephone interview and clinical oral examination) of community-dwelling adults aged 45-64 and ≥65 living in Nova Scotia, Canada was conducted. RESULTS: The survey response rate was 21% for the interview and 13.5% for the examination. A total of 1141 participants completed one or both components of the survey. Both age groups had higher levels of education than the target population; the age 45-64 sample also had a higher proportion of females and lower levels of employment than the target population. Completers (participants who completed interview and examination) were compared with partial completers (who completed only the interview), and stepwise logistic regression was performed to examine predictors of completion. Identified predictors were as follows: not working, post-secondary education and frequent dental visits. CONCLUSION: Recruitment, communications and logistics present challenges in conducting a province-wide survey. Identification of employment, education and dental visit frequency as predictors of survey participation provide insight into possible non-response bias and suggest potential for underestimation of oral disease prevalence in this and similar surveys. This potential must be considered in analysis and in future recruitment strategies.

Compaction strategies for modifying the drug delivery capabilities of gelled calcium polyphosphate matrices.

Calcium polyphosphates (CPPs) have shown potential as drug delivery matrices, particularly in treating bone-related chronic diseases such as osteomyelitis, where maintenance of sufficient bactericidal concentrations at the infected bone site is essential. The objective of this study was to incorporate an additional compaction step as part of a gelling protocol to optimize CPP matrix properties while enhancing their drug delivery capabilities. Vancomycin-loaded CPP powders were produced using a previously established gelling and drying protocol, G1, and then subsequently compacted at prescribed levels (30, 113 or 452MPa) before subjecting to an additional gelling and drying protocol (G2). The resulting G2 disks were found to be more homogeneous and dense (p=0.0013) when compared with corresponding G1 disks, though increases in matrix density did not translate into subsequent increases in tensile strength. The compaction regelling protocol did, however, eliminate the burst release phenomena observed with the G1 disks and further extended the release of vancomycin into a clinically acceptable therapeutic range of 3weeks. These changes were associated with the increase in visual homogeneity, the increase in density and a more homogenous dispersion of vancomycin within the G2 disks. The ability to modulate this release profile to a limited extent by altering compaction stress, particle size distribution and regelling time was also demonstrated. Overall, the compaction regelling protocol described here, when used in conjunction with an initial gelling step to achieve matrix drug loading, enhances the flexibility and long-term drug delivery capability of this CPP matrix.

Measurement of fluid ingress into calcium polyphosphate bioceramics using nuclear magnetic resonance microscopy.

A MR microscopy experiment is developed and used to characterize fluid ingress and microstructural transformation in degradable calcium polyphosphate (CPP) bioceramics. High-resolution (49microm) maps of fluid density and spin-lattice relaxation rate were obtained as a function of time for CPP immersed in phosphate buffered saline. These results demonstrate clear differences in fluid transport rates and solid matrix microstructure in two differing CPP formulations. CPP has been proposed as a potential implantable device for the delivery of pharmaceuticals, and the magnetic resonance imaging (MRI) data are used in conjunction with previously reported bulk elution results to develop a hypothesis explaining microstructural evolution in these materials. This type of non-destructive evaluation of the structure-transport of fluids in CPP is important to improved design of these functionalized biomaterials for long-term, localized delivery of sustained levels of therapeutic agents.

Fibroblastic interactions with high-porosity Ti-6Al-4V metal foam.

A novel metallic Ti-6Al-4V foam in development at the National Research Council of Canada was investigated for its ability to foster cell attachment and growth using a fibroblast cell culture model. The foam was manufactured via a powder metallurgical process that could produce interconnected porosity greater than 70%. Cell attachment was assessed after 6 and 24 h, while proliferation was examined after 3 and 7 days. Ingrown fibroblasts displayed a number of different morphologies; some fibroblasts were spread thinly in close apposition with the irregular surface, or more often had several anchorage points and extended in three dimensions as they spanned pore space. It was also demonstrated that fibroblasts were actively migrating through the porous scaffold over a 14-day period. In a 60-day extended culture, fibroblasts were bridging and filling macropores and had extensively infiltrated the foams. Overall, it was established that this foam was supportive of cell attachment and proliferation, migration through the porous network, and that it was capable of sustaining a large cell population.

The effect of processing on the structural characteristics of vancomycin-loaded amorphous calcium phosphate matrices.

Calcium polyphosphate antibiotic delivery matrices were prepared using a unique processing technique involving the exposure of calcium polyphosphate pastes to high humidity for 0, 5, 24 or 48 h to induce gelling. Subsequently, samples were dried for a minimum of 24 h. The mild conditions associated with matrix fabrication readily allowed for vancomycin incorporation within an environment that did not disrupt antibiotic activity. While reproducible from a processing standpoint, the gelling and drying process did contribute to a decrease in matrix tensile strength and the formation of significant pores near the surface of the matrices. Generally, the core of the gelled matrices appeared to be denser than their non-gelled counterparts. The degree of phosphate chain lysis during the gelling and drying stages was quantified using solution 31P nuclear magnetic resonance (NMR) spectroscopy. Both NMR and Raman spectroscopy indicated that the presence of vancomycin did not appreciably alter the matrix formation process. The ability to incorporate clinically relevant levels of antibiotic within this degradable bone substitute matrix suggests the potential of this approach for creating a localized antibiotic delivery system to treat osteomyelitis infections.

Electrochemically assisted co-precipitation of protein with calcium phosphate coatings on titanium alloy.

A bovine serum albumin protein-containing calcium phosphate coating (BSA/brushite) was prepared by electrochemically assisted co-precipitation onto a hydroxyapatite (HA) coated Ti-6Al-4V surface. Electrochemically assisted co-precipitation of BSA/brushite coatings onto HA resulted in a 70-fold increase in BSA inclusion compared to simple adsorption, and was subsequently released by a slower mechanism (15% loss over 70 h). Thus, this electrochemically assisted co-precipitation technique provides an efficient method of protein incorporation at physiological temperature, with a potential for sustained release of therapeutic agents as may be required for metallic implant fixation.