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Calcium phosphate (CaP) biomaterials are amongst the most widely used synthetic bone graft substitutes, owing to their chemical similarities to the mineral part of bone matrix and off-the-shelf availability. However, their ability to regenerate bone in critical-sized bone defects has remained inferior to the gold standard autologous bone. Hence, there is a need for methods that can be employed to efficiently produce CaPs with different properties, enabling the screening and consequent fine-tuning of the properties of CaPs towards effective bone regeneration. To this end, we propose the use of droplet microfluidics for rapid production of a variety of CaP microparticles. Particularly, this study aims to optimize the steps of a droplet microfluidic-based production process, including droplet generation, in-droplet CaP synthesis, purification and sintering, in order to obtain a library of CaP microparticles with fine-tuned properties. The results showed that size-controlled, monodisperse water-in-oil microdroplets containing calcium- and phosphate-rich solutions can be produced using a flow-focusing droplet-generator microfluidic chip. We optimized synthesis protocols based on in-droplet mineralization to obtain a range of CaP microparticles without and with inorganic additives. This was achieved by adjusting synthesis parameters, such as precursor concentration, pH value, and aging time, and applying heat treatment. In addition, our results indicated that the synthesis and fabrication parameters of CaPs in this method can alter the microstructure and the degradation behavior of CaPs. Overall, the results highlight the potential of the droplet microfluidic platform for engineering CaP microparticle biomaterials with fine-tuned properties.

β-Tricalcium Phosphate (β-TCP), one of the most used bone graft substitutes, may contain up to 5 wt% foreign phase according to standards. Typical foreign phases include β-calcium pyrophosphate (β-CPP) and hydroxyapatite (HA). Currently, the effect of small amounts of impurities on β-TCP resorption is unknown. This is surprising since pyrophosphate is a very potent osteoclast inhibitor. The main aim of this study was to assess the effect of small β-CPP fractions (

Introduction: Local application of antibiotics provides high concentrations at the site of interest, with minimal systemic toxicity. Carrier materials might help manage dead space. Calcium sulphate (CaSO4) has a dissolution time that only slightly exceeds the usually recommended duration of systemic antibiotic treatments. This in vitro study evaluates compatibility, release kinetics and antibacterial activity of new combinations of antibiotics with CaSO4 as carrier material.Methods: CaSO4 pellets added with 8% w/w antibiotic powder were exposed once in phosphate-buffered saline (PBS) solution and once in bovine plasma, in an elution experiment run over 6 weeks at 37 °C. Antibiotic elution was examined at various time points. Concentration was measured by liquid chromatography with tandem mass spectrometry. Antimicrobial activity was checked with an agar diffusion test.Results: Piperacillin-tazobactam, ceftazidime, cefepime, and meropenem showed fast reduction of concentration and activity. Flucloxacillin and cefuroxime remained present in relevant concentrations for 4 weeks. Ciprofloxacin, levofloxacin and clindamycin lasted for 6 weeks, but also at cell toxic concentrations. Ceftriaxone showed a near-constant release with only a small reduction of concentration from 130 to 75 mg/l. Elution profiles from PBS and plasma were comparable.Conclusion: CaSO4 provides new possibilities in the local treatment of bone and joint infections. Ceftriaxone appears to be of particular interest in combination with CaSO4. Release persists at clinically promising concentrations, and appears to have a depot-like slow release from CaSO4, with only a small reduction in activity and concentration over 6 weeks. To the best of our knowledge, such a particular persistent release never was described before, for any antibiotic in combination with a carrier material for local application.

We obtain new sufficient conditions for the oscillation of all solutions of first-order delay dynamic equations on arbitrary time scales, hence combining and extending results for corresponding differential and difference equations. Examples, some of which coincide with well-known results on particular time scales, are provided to illustrate the applicability of our results.

Recent studies have revealed that the ability of a calcium phosphate cement paste to harden in a physiological environment without desintegrating into small particles might be a key property to ensure a safe and reliable clinical use of calcium phosphate cements. However, this property called cohesion is not well understood and has not been studied extensively. The goal of the present study was to better understand which factors affect the cohesion of a calcium phosphate paste using the combination of a theoretical and experimental approach. In the theoretical approach, factors expected to influence the paste cohesion such as Van der Waals forces, electrostatic and steric interactions, as well as osmotic effects were listed and discussed. In the experimental approach, a new method to measure the cohesion of a non-setting calcium phosphate paste was presented and used to assess the effects of various factors on this property. The new method allowed a continuous measurement of cohesion and gave reproducible results. The experimental results confirmed the theoretical predictions: an increase of the liquid-to-powder ratio of the paste and of the powder particle size, as well as the addition of citrate ions and in limited cases dissolved xanthan polymer chains reduced the paste cohesion.

The effect of composition on the reactivity of a calcium phosphate cement (CPC) consisting of tricalcium phosphate (TCP)–water mixtures was investigated by isothermal calorimetry at 37 °C. The parameters of interest were the mean particle size of the powder, the use of small amounts of hydroxyapatite nanoparticles as nucleating agent, the phosphate concentration and the pH of the aqueous solution. Interactions of various parameters were also investigated. The results show that the main parameter controlling CPC reactivity was TCP particle size, because in contrast to other parameters it markedly reducedthe total reaction time. [ABSTRACT FROM AUTHOR]

A well-known formula of Bendixson states that solutions of first-order differential equations, as functions of their initial conditions, satisfy a certain partial differential equation. A consequence is Alekseev's nonlinear variation of parameters formula. In this paper, corresponding results are proved for difference equations. To achieve this, use is made of the recently introduced concept of alpha derivatives, rather than of differences or of the usual derivatives. This technique allows the results to be generalized to alpha dynamic equations, which include among others ordinary differential and difference equations. [ABSTRACT FROM AUTHOR]

Building construction takes time, in part because the binding process of cement is based on the slow re-crystallization and precipitation of calcium silicate species. Since the material's reactivity is surface area limited, a reduction in particle size of Portland cements has been used to prepare faster binding formulations. The present work investigates a new and direct, one-step preparation of calcium silicate-based nanoparticles of a typical Portland cement composition by flame spray synthesis. Isothermal calorimetry revealed that the hardening of this new nano-cement corroborated a more than tenfold increase of initial reactivity with different reaction kinetics if compared to conventionally prepared cements. At present, the unfavourably high porosity of nano-cements, however, underlines the need for additional improvements of chemical composition and formulation to make these highly reactive materials applicable to modern construction work, where load-bearing strength is of importance. [ABSTRACT FROM AUTHOR]

Defining the most adequate architecture of a bone substitute scaffold is a topic that has received much attention over the last 40 years. However, contradictory results exist on the effect of grain size and microporosity. Therefore, the aim of this study was to determine the effect of these two factors on the in vivo behaviour of β-tricalcium phosphate (β-TCP) scaffolds. For that purpose, β-TCP scaffolds were produced with roughly the same macropore size (≈ 150 μm), and porosity (≈ 80 %), but two levels of microporosity (low: 10 % / high: ≈ 25 %) and grain size (small: 1.3 μm /large: ≈ 3.3 μm). The sample architecture was characterised extensively using materialography, Hg porosimetry, micro-computed tomography (μCT), and nitrogen adsorption. The scaffolds were implanted for 2, 4 and 8 weeks in a cylindrical 5-wall cancellous bone defect in sheep. The histological, histomorphometrical and μCT analysis of the samples revealed that all four scaffold types were almost completely resorbed within 8 weeks and replaced by new bone. Despite the three-fold difference in microporosity and grain size, very few biological differences were observed. The only significant effect at p < 0.01 was a slightly faster resorption rate and soft tissue formation between 4 and 8 weeks of implantation when microporosity was increased. Past and present results suggest that the biological response of this particular defect is not very sensitive towards physico-chemical differences of resorbable bone graft substitutes. As bone formed not only in the macropores but also in the micropores, a closer study at the microscopic and localised effects is necessary.

Various compositions of synthetic calcium phosphates (CaP) have been proposed and their use has considerably increased over the past decades. Besides differences in physico-chemical properties, resorption and osseointegration, artificial CaP bone graft might differ in their resistance against biofilm formation. We investigated standardised cylinders of 5 different CaP bone grafts (cyclOS, chronOS (both β-TCP (tricalcium phosphate)), dicalcium phosphate (DCP), calcium-deficient hydroxyapatite (CDHA) and α-TCP). Various physico-chemical characterisations e.g., geometrical density, porosity, and specific surface area were investigated. Biofilm formation was carried out in tryptic soy broth (TSB) and human serum (SE) using Staphylococcus aureus (ATCC 29213) and S. epidermidis RP62A (ATCC 35984). The amount of biofilm was analysed by an established protocol using sonication and microcalorimetry. Physico-chemical characterisation showed marked differences concerning macro- and micropore size, specific surface area and porosity accessible to bacteria between the 5 scaffolds. Biofilm formation was found on all scaffolds and was comparable for α-TCP, chronOS, CDHA and DCP at corresponding time points when the scaffolds were incubated with the same germ and/or growth media, but much lower for cyclOS. This is peculiar because cyclOS had an intermediate porosity, mean pore size, specific surface area, and porosity accessible to bacteria. Our results suggest that biofilm formation is not influenced by a single physico-chemical parameter alone but is a multi-step process influenced by several factors in parallel. Transfer from in vitro data to clinical situations is difficult; thus, advocating the use of cyclOS scaffolds over the four other CaP bone grafts in clinical situations with a high risk of infection cannot be clearly supported based on our data.

In the last 15 years, a large number of commercial ceramic-based cements and putties have been introduced as bone graft substitutes. As a result, large efforts have been made to improve our understanding of the specific properties of these materials, such as injectability, cohesion, setting time (for cements), and in vivo properties. The aim of this manuscript is to summarize our present knowledge in the field. Instead of just looking at scientific aspects, industrial needs are also considered, including mixing and delivery, sterilization, and shelf-life.

The biocompatibility and resorption characteristics of β-tricalcium phosphate (β-TCP, Ca3(PO4)2) have made it a coveted alternative for bone grafts. However, the underlying mechanisms governing the biological interactions between β-tricalcium phosphate and osteoclasts remain elusive. It has been speculated that the composition at grain boundaries might vary and affect β-TCP resorption properties. Atom probe tomography (APT) offers a quantitative approach to assess the composition of the grain boundaries, and thus advance our comprehension of the biological responses within the microstructure and chemical composition at the nanoscale. The precise quantitative analysis of chemical composition remains a notable challenge in APT, primarily due to the influence of measurement conditions on compositional accuracy. In this study, we investigated the impact of laser pulse energy on the composition of β-TCP using APT, aiming for the most precise Ca:P ratio and consistent results across multiple analyses performed with different sets of analysis conditions and on two different instruments., Competing Interests: Conflict of Interest The authors declare that they have no competing interest., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Microscopy Society of America.)

β-TCP ceramics are versatile bone substitute materials and show many interactions with cells of the monocyte-macrophage-lineage. The possibility of monocytes entering microporous β-TCP ceramics has however not yet been researched. In this study, we used a model approach to investigate whether monocytes might enter β-TCP, providing a possible explanation for the origin of CD68-positive osteoclast-like giant cells found in earlier works.We used flow chambers to unidirectionally load BC, PRP, or PPP into slice models of either 2 mm or 6 mm β-TCP. Immunofluorescence for CD68 and live/dead staining was performed after the loading process.Our results show that monocytes were present in a relevant number of PRP and BC slices representing the inside of our 2 mm slice model and also present on the actual inside of our 6 mm model. For PPP, monocytes were not found beyond the surface in either model.Our results indicate the possibility of a new and so far neglected constituent in β-TCP degradation, perhaps causing the process of ceramic degradation also starting from inside the ceramics as opposed to the current understanding. We also demonstrated flow chambers as a possible new in vitro model for interactions between blood and β-TCP., (© 2024. The Author(s).)

Fluorescence analysis of β-TCP ceramics is often used to describe cells found on said ceramics. However, we found, to our knowledge, so far undescribed artifacts which might sometimes be hard to differentiate from cells due to shape and fluorescence behavior. We tried prolonged ultrasound washing as well as Technovit 9100 fixation to reduce these artifacts. While untreated dowels showed no reduction in artifacts no matter the further treatment, Technovit fixation reduced the artifacts with even further reduction achieved by mechanical cleaning. As a consequence, scientists working with these dowels and likely even other types should try to avoid creating false positive results by considering the existence of these artifacts, checking additional filters for unusual fluorescence and by reducing them by using Technovit fixation when possible., (© 2024. The Author(s).)

Calcium phosphate (CaP) biomaterials are amongst the most widely used synthetic bone graft substitutes, owing to their chemical similarities to the mineral part of bone matrix and off-the-shelf availability. However, their ability to regenerate bone in critical-sized bone defects has remained inferior to the gold standard autologous bone. Hence, there is a need for methods that can be employed to efficiently produce CaPs with different properties, enabling the screening and consequent fine-tuning of the properties of CaPs towards effective bone regeneration. To this end, we propose the use of droplet microfluidics for rapid production of a variety of CaP microparticles. Particularly, this study aims to optimize the steps of a droplet microfluidic-based production process, including droplet generation, in-droplet CaP synthesis, purification and sintering, in order to obtain a library of CaP microparticles with fine-tuned properties. The results showed that size-controlled, monodisperse water-in-oil microdroplets containing calcium- and phosphate-rich solutions can be produced using a flow-focusing droplet-generator microfluidic chip. We optimized synthesis protocols based on in-droplet mineralization to obtain a range of CaP microparticles without and with inorganic additives. This was achieved by adjusting synthesis parameters, such as precursor concentration, pH value, and aging time, and applying heat treatment. In addition, our results indicated that the synthesis and fabrication parameters of CaPs in this method can alter the microstructure and the degradation behavior of CaPs. Overall, the results highlight the potential of the droplet microfluidic platform for engineering CaP microparticle biomaterials with fine-tuned properties., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Alaoui Selsouli, Rho, Eischen-Loges, Galván-Chacón, Stähli, Viecelli, Döbelin, Bohner, Tahmasebi Birgani and Habibović.)

The recognition and importance of immune cells during bone regeneration, including around bone biomaterials, has led to the development of an entire field termed "osteoimmunology," which focuses on the connection and interplay between the skeletal system and immune cells. Most studies have focused on the "osteogenic" capacity of various types of bone biomaterials, and much less focus has been placed on immune cells despite being the first cell type in contact with implantable devices. Thus, the amount of literature generated to date on this topic makes it challenging to extract needed information. This review article serves as a guide highlighting advancements made in the field of osteoimmunology emphasizing the role of the osteoimmunomodulatory properties of biomaterials and their impact on osteoinduction. First, the various immune cell types involved in bone biomaterial integration are discussed, including the prominent role of osteal macrophages (OsteoMacs) during bone regeneration. Thereafter, key biomaterial properties, including topography, wettability, surface charge, and adsorption of cytokines, growth factors, ions, and other bioactive molecules, are discussed in terms of their impact on immune responses. These findings highlight and recognize the importance of the immune system and osteoimmunology, leading to a shift in the traditional models used to understand and evaluate biomaterials for bone regeneration., (© 2023 The Authors. Periodontology 2000 published by John Wiley & Sons Ltd.)

Bisphosphonates (BP) are anti-resorptive drugs that are widely used to prevent bone loss in osteoporosis. Since inhibition of bone resorption will cause a decrease in bone formation through a process called coupling, it is hypothesized that extended treatment protocols may impair bone healing. In this study, β-tri‑calcium-phosphate (βTCP) ceramics were inserted into critical-size long bone defects in estrogen-deficient mice under BP therapy. The study assessed the benefits of coating the ceramics with Bone Morphogenetic Protein-2 (BMP2) and an engineered BMP2 analogue (L51P) that inactivates BMP antagonists on the healing process, implant resorption, and bone formation. Female NMRI mice (11-12 weeks of age) were ovariectomized ( OVX ) or sham operated. Eight weeks later, after the manifestation of ovariectomy-induced osteoporotic bone changes, BP therapy with Alendronate (ALN) was commenced. After another five weeks, a femoral critical-size defect was generated, rigidly fixed, and βTCP-cylinders loaded with 0.25 μg or 2.5 μg BMP2, 2.5 μg L51P, and 0.25 μg BMP2/2.5 μg L51P, respectively, were inserted. Unloaded βTCP-cylinders were used as controls. Femora were collected six and twelve weeks post-implantation. Histological and micro-computer tomography (MicroCT) evaluation revealed that insertion of cylinders coated with 2.5 μg BMP2 accelerated fracture repair and induced significant bone formation compared to controls (unloaded cylinders or coated with 2.5 μg L51P, 0.25 μg BMP2) already six weeks post-implantation, independent of estrogen-deficiency and BP therapy. The simultaneous administration of BMP2 and L51P (0.25 μg BMP2/2.5 μg L51P) did not promote fracture healing six and twelve weeks post-implantation. Moreover, new bone formation within the critical-size defect was directly linked to the removal of the βTCP-implant in all experimental groups. No evidence was found that long-term therapy with ALN impaired the resorption of the implanted graft. However, osteoclast transcriptome signature was elevated in sham and OVX animals upon treatment with BP, with transcript levels being higher at six weeks than at twelve weeks post-surgery. Furthermore, the transcriptome profile of the developing repair tissue confirmed an accelerated repair process in animals treated with 2.5 μg BMP2 implants. L51P did not increase the bioefficacy of BMP2 in the applied defect model. The present study provides evidence that continuous administration of BP does not inhibit implant resorption and does not alter the kinetics of the healing process of critical-size long bone defects. Furthermore, the BMP2 variant L51P did not enhance the bioefficacy of BMP2 when applied simultaneously to the femoral critical-size defect in sham and OVX mice., Competing Interests: None., (© 2024 The Authors.)

One of the most widely used materials for bone graft substitution is β-Tricalcium phosphate (β-TCP; β-Ca 3 (PO 4 ) 2 ). β-TCP is typically produced by sintering in air or vacuum. During this process, evaporation of phosphorus (P) species occurs, leading to the formation of a calcium-rich alkaline layer. It was recently shown that the evaporation of P species could be prevented by co-sintering β-TCP with dicalcium phosphate (DCPA; CaHPO 4 ; mineral name: monetite). The aim of this study was to see how a change of sintering atmosphere could affect the physico-chemical and biological properties of β-TCP. For this purpose, three experimental groups were considered: β-TCP cylinders sintered in air and subsequently polished to remove the surface layer (control group); the same polished cylinders after subsequent annealing at 500 °C in air to generate a calcium-rich alkaline layer (annealed group); and finally, β-TCP cylinders sintered in a monetite-rich atmosphere and subsequently polished (monetite group). XPS analysis confirmed that cylinders from the annealed group had a significantly higher Ca/P molar ratio at their surface than that of the control group while this ratio was significantly lower for the cylinders from the monetite group. Sintering β-TCP in the monetite-rich atmosphere significantly reduced the grain size and increased the density. Changes of surface composition affected the activity of osteoclasts seeded onto the surfaces, since annealed β-TCP cylinders were significantly less resorbed than β-TCP cylinders sintered in the monetite-rich atmosphere. This suggests that an increase of the surface Ca/P molar ratio leads to a decrease of osteoclastic resorption. STATEMENT OF SIGNIFICANCE: Minimal changes of surface and bulk (< 1%) composition have major effects on the ability of osteoclasts to resorb β-tricalcium phosphate (β-TCP), one of the most widely used ceramics for bone substitution. The results presented in this study are thus important for the calcium phosphate community because (i) β-TCP may have up to 5% impurities according to ISO and ASTM standards and still be considered to be "pure β-TCP", (ii) β-TCP surface properties are generally not considered during biocompatibility assessment and (iii) a rationale can be proposed to explain the various inconsistencies reported in the literature on the biological properties of β-TCP., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The aim of this study was to produce a composite of microporous β-TCP filled with alginate-gelatin crosslinked hydrogel, clindamycin and bone morphogenetic protein (BMP-2) to prolong the drug-release behaviour for up to 28 days. The most promising alginate-di-aldehyde(ADA)-gelatin gel for drug release from microcapsules was used to fill microporous β-TCP ceramics under directional flow in a special loading chamber. Dual release of clindamycin and BMP-2 was measured on days 1, 2, 3, 6, 9, 14, 21 and 28 by high performance liquid chromatography (HPLC) and enzyme-linked immunosorbent assay (ELISA). After release, the microbial efficacy of the clindamycin was checked and the biocompatibility of the composite was tested in cell culture. Clindamycin and the model substance FITC-protein A were released from microcapsules over 28 days. The clindamycin burst release was 43 ± 1%. For the loaded ceramics, a clindamycin release above the minimal inhibitory concentration (MIC) until day 9 and a burst release of 90.56 ± 2.96% were detected. BMP-2 was released from the loaded ceramics in low concentrations over 28 days. The release of active substances from β-TCP and hydrogel have already been extensively studied. Directional flow loading is a special procedure in which the ceramic could act as a stabilizer in the bone and, as a biodegradable system, enables a single-stage surgical procedure. Whether ADA-gelatin gel is suitable for this procedure as a more biodegradable alternative to pure alginate or whether a dual release is possible in this composite has not yet been investigated., (© 2023. The Author(s).)

Background: Low-field magnetic resonance imaging (MRI) has gained increasing importance to monitor equine tendon lesions. Comparing results between studies and cases is hampered, because image analysis approaches vary strongly. This study aimed to improve reliability, comparability and time efficiency of quantitative MRI image analysis., Methods: Induced tendon lesions were studied over a 24-week period with 10 follow-up MRI examinations. Signal intensities (SIs) of tendons, tendon lesions, cortical bone and background, as well as lesion cross-sectional areas (CSAs) were measured. Lesion SI standardisation with different formulas was evaluated, using histological findings as reference. Different types of region of interest (ROI) for lesion SI measurement were compared. Lesion CSA measurement at different levels was evaluated, using the calculated total lesion volume as reference. Subjective lesion identification and manual CSA and SI measurements were compared to an automated, algorithm-based approach., Results: Lesion SI standardised using a quotient of lesion and background or cortical bone SI, correlated best with histologically determined lesion severity. Lesion SI in circular ROIs correlated strongly with lesion SI in free-hand whole-lesion ROIs. The level of the maximum lesion CSA shifted over time; the CSA maximum correlated strongly with lesion volume. In sequences with short acquisition time, algorithm-based automated lesion detection showed almost perfect agreement with subjective lesion identification. Automated measurement of CSA and SI was also feasible, with stronger correlation and better agreement with the manually obtained data for the SI than for the CSA., Conclusion: Our study may provide guidance for MRI image analysis of tendon healing. Reliable image analysis can be performed time-efficiently, particularly regarding lesion SI quantification., Competing Interests: The authors declare they have no conflicts of interest., (© 2023 The Authors. Veterinary Record Open published by John Wiley & Sons Ltd on behalf of British Veterinary Association.)

In this work, we formulate the Beverton-Holt model on isolated time scales and extend existing results known in the discrete and quantum calculus cases. Applying a recently introduced definition of periodicity for arbitrary isolated time scales, we discuss the effects of periodicity onto a population modeled by a dynamic version of the Beverton-Holt equation. The first main theorem provides conditions for the existence of a unique ω -periodic solution that is globally asymptotically stable, which addresses the first Cushing-Henson conjecture on isolated time scales. The second main theorem concerns the generalization of the second Cushing-Henson conjecture. It investigates the effects of periodicity by deriving an upper bound for the average of the unique periodic solution. The obtained upper bound reveals a dependence on the underlying time structure, not apparent in the classical case. This work also extends existing results for the Beverton-Holt model in the discrete and quantum cases, and it complements existing conclusions on periodic time scales. This work can furthermore guide other applications of the recently introduced periodicity on isolated time scales.

Heterotopic ossification (HO) is a condition triggered by an injury leading to the formation of mature lamellar bone in extraskeletal soft tissues. Despite being a frequent complication of orthopedic and trauma surgery, brain and spinal injury, the etiology of HO is poorly understood. The aim of this study is to evaluate the hypothesis that a sustained local ionic homeostatic imbalance (SLIHI) created by mineral formation during tissue calcification modulates inflammation to trigger HO. This evaluation also considers the role SLIHI could play for the design of cell-free, drug-free osteoinductive bone graft substitutes. The evaluation contains five main sections. The first section defines relevant concepts in the context of HO and provides a summary of proposed causes of HO. The second section starts with a detailed analysis of the occurrence and involvement of calcification in HO. It is followed by an explanation of the causes of calcification and its consequences. This allows to speculate on the potential chemical modulators of inflammation and triggers of HO. The end of this second section is devoted to in vitro mineralization tests used to predict the ectopic potential of materials. The third section reviews the biological cascade of events occurring during pathological and material-induced HO, and attempts to propose a quantitative timeline of HO formation. The fourth section looks at potential ways to control HO formation, either acting on SLIHI or on inflammation. Chemical, physical, and drug-based approaches are considered. Finally, the evaluation finishes with a critical assessment of the definition of osteoinduction. STATEMENT OF SIGNIFICANCE: The ability to regenerate bone in a spatially controlled and reproducible manner is an essential prerequisite for the treatment of large bone defects. As such, understanding the mechanism leading to heterotopic ossification (HO), a condition triggered by an injury leading to the formation of mature lamellar bone in extraskeletal soft tissues, would be very useful. Unfortunately, the mechanism(s) behind HO is(are) poorly understood. The present study reviews the literature on HO and based on it, proposes that HO can be caused by a combination of inflammation and calcification. This mechanism helps to better understand current strategies to prevent and treat HO. It also shows new opportunities to improve the treatment of bone defects in orthopedic and dental procedures., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

We derive a discrete predator-prey model from first principles, assuming that the prey population grows to carrying capacity in the absence of predators and that the predator population requires prey in order to grow. The proposed derivation method exploits a technique known from economics that describes the relationship between continuous and discrete compounding of bonds. We extend standard phase plane analysis by introducing the next iterate root-curve associated with the nontrivial prey nullcline. Using this curve in combination with the nullclines and direction field, we show that the prey-only equilibrium is globally asymptotic stability if the prey consumption-energy rate of the predator is below a certain threshold that implies that the maximal rate of change of the predator is negative. We also use a Lyapunov function to provide an alternative proof. If the prey consumption-energy rate is above this threshold, and hence the maximal rate of change of the predator is positive, the discrete phase plane method introduced is used to show that the coexistence equilibrium exists and solutions oscillate around it. We provide the parameter values for which the coexistence equilibrium exists and determine when it is locally asymptotically stable and when it destabilizes by means of a supercritical Neimark-Sacker bifurcation. We bound the amplitude of the closed invariant curves born from the Neimark-Sacker bifurcation as a function of the model parameters., (© 2022. The Author(s), under exclusive licence to Society for Mathematical Biology.)

The aim of this work was to establish an organ model for staphylococcal infection of human bone samples and to investigate the influence and efficacy of a microporous β-tricalcium phosphate ceramic (β-TCP, RMS Foundation) loaded with hydrogels (alginate, alginate-di-aldehyde (ADA)-gelatin) and clindamycin on infected human bone tissue over a period of 28 days. For this purpose, human tibia plateaus, collected during total knee replacement surgery, were used as a source of bone material. Samples were infected with S. aureus ATCC29213 and treated with differently loaded β-TCP composites (alginate +/- clindamycin, ADA-gelatin +/- clindamycin, unloaded). The loading of the composites was carried out by means of a flow chamber. The infection was observed for 28 days, quantifying bacteria in the medium and the osseus material on day 1, 7, 14, 21 and 28. All samples were histologically processed for bone vitality evaluation. Bone infection could be consistently performed within the organ model. In addition, a strong reduction in bacterial counts was recorded in the groups treated with ADA-gelatin + clindamycin and alginate + clindamycin, while the bacterial count in the control groups remained constant. No significant differences between groups could be observed in the number of lacunae filled with osteocytes suggesting no differences in bone vitality among groups. In an ex-vivo human bone infection model, over a period of 28 days bacterial growth could be reduced by treatment with ADA-Gel + CLI and ALG + CLI -releasing β-TCP composites. This could be relevant for its clinical use. Further work will be necessary to improve the loading of β-TCP and the bone infection organ model itself. STATEMENT OF SIGNIFICANCE: The common treatment of bone infections is debridement and systemic administration of antibiotics. In some cases, antibiotic-containing carriers are already used, but these must be removed again. Our work is intended to show another treatment option. The scaffold we have developed, made of a calcium phosphate ceramic and a hydrogel as the active substance carrier, can, in addition to releasing the active substance, also assume a load-bearing function of the bone and is biodegradable. In addition, the model we developed can also be used for the analysis and treatment of bone infections other than those of the musculoskeletal system. More importantly, it can also serve as a substitute for previously used animal experiments., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2022. Published by Elsevier Ltd.)