Biocompatibility and Biological Performance Evaluation of Additive-Manufactured Bioabsorbable Iron-Based Porous Suture Anchor in a Rabbit Model.

This study evaluated the biocompatibility and biological performance of novel additive-manufactured bioabsorbable iron-based porous suture anchors (iron_SAs). Two types of bioabsorbable iron_SAs, with double- and triple-helical structures (iron_SA_2_helix and iron_SA_3_helix, respectively), were compared with the synthetic polymer-based bioabsorbable suture anchor (polymer_SAs). An in vitro mechanical test, MTT assay, and scanning electron microscope (SEM) analysis were performed. An in vivo animal study was also performed. The three types of suture anchors were randomly implanted in the outer cortex of the lateral femoral condyle. The ultimate in vitro pullout strength of the iron_SA_3_helix group was significantly higher than the iron_SA_2_helix and polymer_SA groups. The MTT assay findings demonstrated no significant cytotoxicity, and the SEM analysis showed cells attachment on implant surface. The ultimate failure load of the iron_SA_3_helix group was significantly higher than that of the polymer_SA group. The micro-CT analysis indicated the iron_SA_3_helix group showed a higher bone volume fraction (BV/TV) after surgery. Moreover, both iron SAs underwent degradation with time. Iron_SAs with triple-helical threads and a porous structure demonstrated better mechanical strength and high biocompatibility after short-term implantation. The combined advantages of the mechanical superiority of the iron metal and the possibility of absorption after implantation make the iron_SA a suitable candidate for further development.

IL (Interleukin)-33 Suppresses Abdominal Aortic Aneurysm by Enhancing Regulatory T-Cell Expansion and Activity.

Objective- Inflammation occurs during the progression of abdominal aortic aneurysm (AAA). IL (interleukin)-33 is a pleiotropic cytokine with multiple immunomodulatory effects, yet its role in AAA remains unknown. Approach and Results- Immunoblot, immunohistochemistry, and immunofluorescent staining revealed increased IL-33 expression in adventitia fibroblasts from mouse AAA lesions. Daily intraperitoneal administration of recombinant IL-33 or transgenic IL-33 expression ameliorated periaorta CaPO4 injury- and aortic elastase exposure-induced AAA in mice, as demonstrated by blunted aortic expansion, reduced aortic wall elastica fragmentation, enhanced AAA lesion collagen deposition, attenuated T-cell and macrophage infiltration, reduced inflammatory cytokine production, skewed M2 macrophage polarization, and reduced lesion MMP (matrix metalloproteinase) expression and cell apoptosis. Flow cytometry analysis, immunostaining, and immunoblot analysis showed that exogenous IL-33 increased CD4+Foxp3+ regulatory T cells in spleens, blood, and aortas in periaorta CaPO4-treated mice. Yet, ST2 deficiency muted these IL-33 activities. Regulatory T cells from IL-33-treated mice also showed significantly stronger activities in suppressing smooth muscle cell inflammatory cytokine and chemokine expression, macrophage MMP expression, and in increasing M2 macrophage polarization than those from vehicle-treated mice. In contrast, IL-33 failed to prevent AAA and lost its beneficial activities in CaPO4-treated mice after selective depletion of regulatory T cells. Conclusions- Together, this study established a role of IL-33 in protecting mice from AAA formation by enhancing ST2-dependent aortic and systemic regulatory T-cell expansion and their immunosuppressive activities.

Use of mechanistic information to derive chemical-specific adjustment factors - Refinement of risk assessment.

When extrapolating data from animal toxicological studies a default factor (dUF) of 100 is applied to derive a heath based guidance value. The UF takes into account the interspecies differences (ID) and the intraspecies variability (IV). When re-evaluating the safety of phosphates used as food additives nephrocalcinosis was identified as the critical endpoint. The underlying mechanism for nephrocalcinosis was attributed to the precipitation of calcium phosphate in the kidney, depending on its solubility, irrespective of the species and the population. Based on the mechanism, the volume of primary urine, for which the glomerular filtration rate (GFR) was used as a proxy, was considered to be the only parameter relevant for ID and IV. Median value of GFR in rats was 4.0 ml/min/kg bw. In humans it was 1.6 ml/min/kg bw in healthy adults and 0.9 in elderly. These values were calculated from the distribution of the GFR data from 8 studies in rats (n = 191), 16 studies in adults (n = 1540) and 5 studies in elderly (n = 2608). Multiplying the distribution of the ratio rat/healthy humans (ID) with the distribution of the ratio healthy humans/elderly human (IV) resulted in a phosphate specific factor of 4.5 (3.3-6.7) (median; 25th - 75th percentile).

Physico-Mechanical Properties of HA/TCP Pellets and Their Three-Dimensional Biological Evaluation In Vitro.

The use of bioceramics, especially the combination of hydroxyapatite (HA) and ß-tricalcium phosphate (ß-TCP), as a three-dimensional scaffold in bone engineering is essential because together these elements constitute 60% of the bone content. Different ratios of HA and ß-TCP were previously tested for their ability to produce suitable bioceramic scaffolds, which must be able to withstand high mechanical load. In this study, two ratios of HA/TCP (20:80 and 70:30) were used to create pellets, which then were evaluated in vitro to identify any adverse effects of using the material in bone grafting. Diametral tensile strength (DTS) and density testing was conducted to assess the mechanical strength and porosity of the pellets. The pellets then were tested for their toxicity to normal human fibroblast cells. In the toxicity assay, cells were incubated with the pellets for 3 days. At the end of the experiment, cell morphological changes were assessed, and the absorbance was read using PrestoBlue Cell Viability Reagent™. An inversely proportional relationship between DTS and porosity percentage was detected. Fibroblasts showed normal cell morphology in both treatments, which suggests that the HA/TCP pellets were not toxic. In the osteoblast cell attachment assay, cells were able to attach to the surface of both ratios, but cells were also able to penetrate inside the scaffold of the 70:30 pellets. This finding suggests that the 70:30 ratio had better osteoconduction properties than the 20:80 ratio.

In vitro cytotoxicity of calcium phosphate cement reinforced with multiwalled carbon nanotubes.

The in vitro cytotoxicity of both the multiwalled carbon nanotubes (MWCNT) in suspension with culture medium and the tetracalcium phosphate/monetite cement with addition of 0.8 wt% of MWCNTs on fibroblasts and osteoblasts were studied. The cytotoxicity was evaluated by MTS test (formazan) and live/dead staining. No cytotoxicity of MWCNT extract was measured contrary to about 60% reduction in proliferation of fibroblasts in MWCNT suspension as compared with negative control. The several contact cytotoxicity of MWCNT composite cement surfaces on seeded cells was demonstrated by MTS test and live/dead staining of damaged fibroblasts and dead osteoblasts after 72 h of culture. The detailed microstructure analysis showed a significant refinement of the surface texture due to the formation of thin needle-like hydroxyapatite particles on MWCNTs and this effect could be responsible for cytotoxicity of composites.

Investigation of the Genotoxicity of Aluminum Oxide, ß-Tricalcium Phosphate, and Zinc Oxide Nanoparticles In Vitro.

The aim of this study was to investigate the genotoxicity of aluminum oxide (Al2O3), ß-tricalcium phosphate (ß-TCP) (Ca3(PO4)2), and zinc oxide (ZnO) nanoparticles (NPs) that were 4.175, 9.058, and 19.8 nm sized, respectively, on human peripheral blood lymphocytes using micronucleus (MN) and chromosome aberration (CA) techniques. Aluminum oxide and ß-TCP NPs did not show genotoxic effects on human peripheral blood cultures in vitro, even at the highest concentrations; therefore, these materials may be suitable for use as biocompatible materials. It was observed that, even at a very low dose (≥12.5 ppm), ZnO NPs had led to genotoxicity. In addition, at high concentrations (500 ppm and above), ZnO NPs caused mortality of lymphocytes. For these reasons, it was concluded that ZnO NPs are not appropriate for using as a biocompatible biomaterial.

Comparative Evaluation of Cytotoxicity and Genotoxicity of Two Bioceramic Sealers on Fibroblast Cell Line: An in vitro Study.

AIM: The purpose of this study was to evaluate and compare the cytotoxicity and genotoxicity of two bioceramic root canal sealers: EndoSequence BC and iRoot SP with zinc oxide eugenol sealers on fibroblast cell line. MATERIALS AND METHODS: The sealers tested were zinc oxide eugenol, EndoSequence BC, and iRoot SP. Each material was mixed according to the manufacturer's instructions and mounted into sterile polyethylene color-coded rings, for cytotoxicity and genotoxicity evaluation. After 48 hours, the set materials were transferred to previously marked wells and cytotoxicity evaluation to L929 murine fibroblast cells was done by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. The percentages of viable cells were then calculated and values were statistically analyzed by Kruskal-Wallis test. The evaluation of genotoxicity of the materials to L929 murine fibroblast cells was carried out by Comet assay. To quantify deoxyribonucleic acid (DNA) damage, the following comet parameters were evaluated in the assay using Comet scoring software: tail length, tail moment, and Olive moment. The values were statistically analyzed using Kruskal-Wallis test with a significance value set to p < 0.05. RESULTS: The results of the study showed that both cytotoxicity and genotoxicity evaluation by MTT assay and Comet assay can be done on L929 murine fibroblast cell line. Among the three tested materials, zinc oxide eugenol showed maximum cytotoxicity to the cells (30.64% viable cells), followed by EndoSequence BC (71.33% viable cells) and iRoot SP (75.11% viable cells). The evaluation of DNA damage by genotoxicity assessment showed iRoot SP to be least genotoxic followed closely by EndoSequence BC. Zinc oxide eugenol was genotoxic and induced more DNA damage on the fibroblast cell line studied. The statistical analyses for both the assays were nonsignificant. CONCLUSION: All the three tested sealers showed varying degrees of cytotoxicity and genotoxicity while using fibro-blast cell line. Zinc oxide eugenol was most toxic in both the assays and iRoot SP showed least toxicity, followed closely by EndoSequence BC.

Activation of Human Transient Receptor Potential Melastatin-8 (TRPM8) by Calcium-Rich Particulate Materials and Effects on Human Lung Cells.

To better understand how adverse health effects are caused by exposure to particulate materials, and to develop preventative measures, it is important to identify the properties of particles and molecular targets that link exposure with specific biologic outcomes. Coal fly ash (CFA) is a by-product of coal combustion that can affect human health. We report that human transient receptor potential melastatin-8 (TRPM8) and an N-terminally truncated TRPM8 variant (TRPM8-Δ801) are activated by CFA and calcium-rich nanoparticles and/or soluble salts within CFA. TRPM8 activation by CFA was potentiated by cold temperature involving the phosphatidylinositol 4,5-bisphosphate binding residue (L1008), but was independent of the icilin and menthol binding site residue Y745 and, essentially, the N-terminal amino acids 1-800. CFA, calcium nanoparticles, and calcium salts also activated transient receptor potential vanilloid-1 (TRPV1) and transient receptor potential ankyrin-1 (TRPA1), but not TRPV4. CFA treatment induced CXCL1 and interleukin-8 mRNA in BEAS-2B and primary human bronchial epithelial cells through activation of both TRPM8 and TRPV1. However, neither mouse nor rat TRPM8 was activated by these materials, and Trpm8 knockout had no effect on cytokine induction in the lungs of CFA-instilled mice. Amino acids S921 and S927 in mouse Trpm8 were identified as important for the lack of response to CFA. These results imply that TRPM8, in conjunction with TRPV1 and TRPA1, might sense selected forms of inhaled particulate materials in human airways, shaping cellular responses to these materials, and improving our understanding of how and why certain particulate materials elicit different responses in biologic systems, affecting human health.

Thrombospondin-1 (TSP1) contributes to the development of vascular inflammation by regulating monocytic cell motility in mouse models of abdominal aortic aneurysm.

RATIONALE: Histological examination of abdominal aortic aneurysm (AAA) tissues demonstrates extracellular matrix destruction and infiltration of inflammatory cells. Previous work with mouse models of AAA has shown that anti-inflammatory strategies can effectively attenuate aneurysm formation. Thrombospondin-1 is a matricellular protein involved in the maintenance of vascular structure and homeostasis through the regulation of biological functions, such as cell proliferation, apoptosis, and adhesion. Expression levels of thrombospondin-1 correlate with vascular disease conditions. OBJECTIVE: To use thrombospondin-1-deficient (Thbs1(-/-)) mice to test the hypothesis that thrombospondin-1 contributes to pathogenesis of AAAs. METHODS AND RESULTS: Mouse experimental AAA was induced through perivascular treatment with calcium phosphate, intraluminal perfusion with porcine elastase, or systemic administration of angiotensin II. Induction of AAA increased thrombospondin-1 expression in aortas of C57BL/6 or apoE-/- mice. Compared with Thbs1(+/+) mice, Thbs1(-/-) mice developed significantly smaller aortic expansion when subjected to AAA inductions, which was associated with diminished infiltration of macrophages. Thbs1(-/-) monocytic cells had reduced adhesion and migratory capacity in vitro compared with wild-type counterparts. Adoptive transfer of Thbs1(+/+) monocytic cells or bone marrow reconstitution rescued aneurysm development in Thbs1(-/-) mice. CONCLUSIONS: Thrombospondin-1 expression plays a significant role in regulation of migration and adhesion of mononuclear cells, contributing to vascular inflammation during AAA development.

Basic calcium phosphate crystals and osteoarthritis pathogenesis: novel pathways and potential targets.

PURPOSE OF REVIEW: Basic calcium phosphate (BCP) crystals have long been associated with the pathogenesis of osteoarthritis. As our knowledge concerning BCP crystals in osteoarthritis expands, so does the potential to develop targeted therapies. The present review discusses recent advances in this field and attempts to summarize our current understanding regarding the role of BCP crystals in osteoarthritis pathogenesis. RECENT FINDINGS: BCP crystals injected into the knees of mice induce osteoarthritis-like changes, further evidence of their pathogenic properties. Interleukin-6 has emerged as a key cytokine involved in BCP crystal-induced inflammation that could represent a potential therapeutic target. The role of BCP crystal-induced osteoclastogenesis has also recently been explored and may also hold the key to future targeted therapies. Although tools to detect BCP crystals remain limited, dual energy computerized tomography scanning has emerged as a useful noninvasive means of quantifying intra-articular calcium crystal deposition. SUMMARY: BCP crystals can activate a number of inflammatory pathways which in turn may lead to cartilage degradation and osteoarthritis. Understanding of these pathways may ultimately yield targeted therapies for osteoarthritis, for which none currently exists.

Preparation of calcium phosphate cement and polymethyl methacrylate for biological composite bone cements.

BACKGROUND: We studied the biological safety, biomechanics, and tissue compatibility of calcium phosphate cement and Polymethyl Methacrylate composite bone cement mixed in different ratios. MATERIAL/METHODS: CPC and PMMA were mixed in different ratios (3:1, 2:1, 1:1, 1:2, 1:5, 1:10, 1:15, and 1:20). PMMA solvent is a general solvent containing a dissolved preparation of the composite bone cement specific to a given specimen to determine biological safety, biomechanics, and tissue compatibility. RESULTS: The CPC/PMMA (33%) group, CPC/PMMA (50%) group, CPC/PMMA (67%) group, and CPC/PMMA (75%) group were more in line with the composite bone cement without cytotoxicity requirements. The compressive strength of the CPC/PMMA (67%) group and CPC/PMMA (75%) group was 20 Mpa-30 Mpa, while that of the CPC/PMMA (4.8%) group, CPC/PMMA (6.25%) group, CPC/PMMA (9.1%) group, CPC/PMMA (16.7%) group, CPC/PMMA (33%) group, and CPC/PMMA (50%) group was 40 Mpa-70 Mpa. Curing time was longer in the CPC group (more than 11 min) and shorter in the PMMA group (less than 2 min). The results of weight loss rate showed that there were no significant differences between the CPC/PMMA group (4.8%, 6.25%, 9.1%, 16.7%, 33%) and PMMA control group (p>0.05). With the decrease of CPC content, the rate of weight loss gradually decreased. CONCLUSIONS: The CPC/PMMA (50%) group, CPC/PMMA (67%) group, and CPC/PMMA (75%) group provide greater variability and selectivity for the composite bone cement in obtaining better application.

A novel root-end filling material based on hydroxyapatite, tetracalcium phosphate and polyacrylic acid.

AIM: To develop a novel root-end filling material (NRFM) based on hydroxyapatite, tetracalcium phosphate and polyacrylic acid, to determine its chemical composition and to compare its physical properties and cytotoxicity with those of glass-ionomer cement (GIC) and grey Portland cement (GPC). METHODOLOGY: The NRFM was prepared by blending distilled water with powders of hydroxyapatite, tetracalcium phosphate, solid polyacrylic acid, solid citric acid and sodium citrate. Chemical analysis was then performed by X-ray diffraction and Fourier transform infrared spectrophotometry. Physical properties were compared with GIC and GPC regarding setting time, compressive strength, pH value and washout resistance; cytotoxicity was assessed using a transwell cell culture system. RESULTS: (i) The NRFM was primarily composed of HA, calcium polyacrylate and calcium citrate. (ii) The mean and standard deviation setting time of NRFM was 11.0 (0.8) min; its compressive strength was 25.6 (2.7) MPa and 38.2 (5.7) MPa at 1 and 7 days respectively. Its pH value ranged from 6.14 to 8.28 and it remained intact after the washout test, whereas GIC and GPC disintegrated. (iii) Dimethyl-thiazol-diphenyl tetrazolium bromide (MTT) and crystal violet (CV) assay revealed that cell-viability on the NRFM was not significantly different form that of the negative control group after treatment for 24, 48 and 72 h. CONCLUSION: The novel root-end filling material (NRFM) is a promising root-end filling material with good physicochemical properties and low cytotoxicity.

A new HA/TTCP material for bone augmentation: an in vivo histological pilot study in primates sinus grafting.

OBJECTIVE: Synthetic calcium phosphate bone substitutes are widely used in sinus graft procedures due to their osteoconductive and biocompatible properties. Hydroxyapatite (HA), beta-tricalcium phosphate (ß-TCP), and HA/ß-TCP composite are the most applied materials. The aim of this study was to propose a new mineralogical formulation, HA/tetracalcium phosphate (TTCP), as biomaterial for bone regeneration in the maxillary sinus. METHODS: Sinus grafts were performed by using granules of a HA/TTCP blend and a collagen membrane. Bone response at time points of 14 and 17 weeks was histologically evaluated. RESULTS: After 14 weeks of healing, histomorphometric analysis showed the formation of new bone trabeculae among HA/TTCP granules. After 17 weeks, the bone trabeculae were thicker and HA/TTCP granules were still present. Histomorphometric analysis revealed a bone graft contact (BGC) of 64%. CONCLUSIONS: After 17 weeks from implantation, HA/TTCP synthetic bone graft performed very well as osteoconductive material: BGC was found very high, and bone volume and vital bone showed an ideal bone density for implant placement. HA/TTCP granules are accounted for to accelerate new bone formation and to reduce the time needed for the graft healing, thus achieving high quantity of the new bone formed.

[Biocompatibility of polymer-bioglass cement Cortoss®: in vitro test with the MG63 cell model]. / Biokompatibilität von Polymer-Glaskeramik-Zement Cortoss®: In-vitro-Testung mit dem MG63-Zell-Modell.

BACKGROUND: Polymethylmethacrylate (PMMA) cement has been used for fixation of joint replacements for more than 50 years and cement augmentation of vertebrae has become a popular procedure since the first description in 1987. New cements have now been developed which are better suited to the requirements of minimally invasive application techniques for vertebral bodies. The combination of good mechanical properties and biocompatibility is the concern of present research. This study compared the features of a polymer-bioglass cement with a calcium phosphate cement used for vertebral augmentation. METHODS: The human osteoblast-like cell culture MG63 was used to study the polymer-glass ceramic cement Cortoss® and the hydroxyapatite cement Kyphos®. Every 24 h for 5-6 days a defined volume of the culture medium was harvested in the presence of the bone cements and added to 16 cell cultures for each time period. The viability of cells was determined photometrically at 550 nm with the MTT assay and cell morphology was studied using light and electron microscopy. RESULTS: In the presence of the calcium phosphate cement an early and small reduction of cell activity was found compared with the controls. At the end of 1 week the viability parameter improved nearly reaching the control level. Electron microscopy showed crystals with a 3-dimensional shape. The cell cultures with Cortoss® showed no cellular activity and the microscopic examinations were negative. This effect was not different at days 1-5 after polymerization of the cement. CONCLUSIONS: The calcium phosphate cement studied showed a good biocompatibility and allowed morphological signs of apatite formation. At least within the first 5 days the polymer-glass ceramic cement showed a reasonable cytotoxic effect. There was no sign of recovery of cell function within that period. The biocompatibility of the polymer-glass ceramic cement appeared significantly worse compared with the calcium phosphate cement. An ideal composition of biomechanical properties and biocompatibility has not been achieved so far.

[Preparation and in vitro evaluation of pDNA-CaPi-PLGA nanoparticles with a core-shell structure].

To develop a core-shell structure pDNA-CaPi-PLGA nanoparticles (CS-pDNA-CaPi-PLGA-NPs), calcium phosphate-pDNA nano complexes (CaPi-pDNA) were encapsulated inside of PLGA shells. The characteristics of the nanoparticles, including morphology, average particle size, zeta potential, entrapment efficiency, loading efficiency, stability in medium, pDNA protection ability from nuclease degradation, in vitro release, cytotoxicity and cell transfection were investigated and compared with the embedded structured CaPi modified PLGA nanoparticles (embedded-pDNA-CaPi-PLGA-NPs). The results showed that the obtained CS-pDNA-CaPi-PLGA-NPs were spherical in shape with an average particle size of (155 +/- 4.5) nm, zeta potentials of (-0.38 +/- 0.1) mV, entrapment efficiency of (80.56 +/- 2.5)% and loading efficiency of (1.16 +/- 0.04)%. The CS-pDNA-CaPi-PLGA-NPs were stable in the release media and could protect pDNA against nuclease degradation. And they also exhibited sustained release of pDNA in vitro. The highest gene transfection efficiency of the CS-pDNA-CaPi-PLGA-NPs in vitro reached (24.66 +/- 0.46)% (after 72 h transfection), which was significantly higher than that of free pDNA [(0.33 +/- 0.04)%, P < 0.01] and the pDNA-PLGA-NPs [(1.5 +/- 0.07)%, P < 0.01]. Besides, the transfection lasted for longer time than that of embedded-pDNA-CaPi-PLGA-NPs and the cytotoxicity of it was significantly lower than that of PEI (P < 0.01). These results indicate that CS-pDNA-CaPi-PLGA-NPs are a promising non-viral gene vector. Key words: gene delivery system; polylactic-co-glycolic acid; calcium phosphate; nanoparticle

Assessment of physical/mechanical properties and cytotoxicity of dual-cured resin cements containing Sr-bioactive glass nanoparticles and calcium phosphate.

The aim was to develop dual-cured resin cements containing Sr-bioactive glass nanoparticles (Sr-BGNPs; 5 or 10 wt%) and monocalcium phosphate monohydrate (MCPM; 3 or 6 wt%). Effects of additives on degree of monomer conversion (DC), biaxial flexural strength/modulus, shear bond strength (SBS), mass/volume change, color stability, ion release, and cytotoxicity were examined. Controls included material without reactive fillers and Panavia SA Plus (PV). Experimental cements showed higher DC than PV regardless of light activation (p<0.05). Mean SBS and color stability were comparable between experimental cements and PV. Cell viability upon the exposure to sample extracts of experimental cements was 80%-92%. High additive concentrations led to lower strength and modulus than PV (p<0.05). The additives increased mass change, reduced color stability, and promoted ion release. The experimental resin cements demonstrated acceptable mechanical/chemical properties and cytotoxicity. The additives reduced the strength but provided ion release, a desirable action to prevent recurrent caries.

Octacalcium phosphate crystals induce inflammation in vivo through interleukin-1 but independent of the NLRP3 inflammasome in mice.

OBJECTIVE: To determine the mechanisms involved in inflammatory responses to octacalcium phosphate (OCP) crystals in vivo. METHODS: OCP crystal-induced inflammation was monitored using a peritoneal model of inflammation in mice with different deficiencies affecting interleukin-1 (IL-1) secretion (IL-1α(-/-) , IL-1ß(-/-) , ASC(-/-) , and NLRP3(-/-) mice) or in mice pretreated with IL-1 inhibitors (anakinra [recombinant IL-1 receptor antagonist] and anti-IL-1ß). The production of IL-1α, IL-1ß, and myeloid-related protein 8 (MRP-8)-MRP-14 complex was determined by enzyme-linked immunosorbent assay. Peritoneal neutrophil recruitment and cell viability were determined by flow cytometry. Depletion of mast cells or resident macrophages was performed by pretreatment with compound 48/80 or clodronate liposomes, respectively. RESULTS: OCP crystals induced peritoneal inflammation, as demonstrated by neutrophil recruitment and up-modulation of IL-1α, IL-1ß, and MRP-8-MRP-14 complex, to levels comparable with those induced by monosodium urate monohydrate crystals. This OCP crystal-induced inflammation was both IL-1α- and IL-1ß-dependent, as shown by the inhibitory effects of anakinra and anti-IL-1ß antibody treatment. Accordingly, OCP crystal stimulation resulted in milder inflammation in IL-1α(-/-) and IL-1ß(-/-) mice. Interestingly, ASC(-/-) and NLRP3(-/-) mice did not show any alteration in their inflammation status in response to OCP crystals. Depletion of the resident macrophage population resulted in a significant decrease in crystal-induced neutrophil infiltration and proinflammatory cytokine production in vivo, whereas mast cell depletion had no effect. Finally, OCP crystals induced apoptosis/necrosis of peritoneal cells in vivo. CONCLUSION: These data indicate that macrophages, rather than mast cells, are important for initiating and driving OCP crystal-induced inflammation. Additionally, OCP crystals induce IL-1-dependent peritoneal inflammation without requiring the NLRP3 inflammasome.

Corrosion Resistance and Cytocompatibility of Magnesium-Calcium Alloys Modified with Zinc- or Gallium-Doped Calcium Phosphate Coatings.

In orthopedic surgery, metals are preferred to support or treat damaged bones due to their high mechanical strength. However, the necessity for a second surgery for implant removal after healing creates problems. Therefore, biodegradable metals, especially magnesium (Mg), gained importance, although their extreme susceptibility to galvanic corrosion limits their applications. The focus of this study was to control the corrosion of Mg and enhance its biocompatibility. For this purpose, surfaces of magnesium-calcium (MgCa1) alloys were modified with calcium phosphate (CaP) or CaP doped with zinc (Zn) or gallium (Ga) via microarc oxidation. The effects of surface modifications on physical, chemical, and mechanical properties and corrosion resistance of the alloys were studied using surface profilometry, goniometry, scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), nanoindentation, and electrochemical impedance spectroscopy (EIS). The coating thickness was about 5-8 µm, with grain sizes of 43.1 nm for CaP coating and 28.2 and 58.1 nm for Zn- and Ga-doped coatings, respectively. According to EIS measurements, the capacitive response (Yc) decreased from 11.29 to 8.72 and 0.15 Ω-1 cm-2 sn upon doping with Zn and Ga, respectively. The Ecorr value, which was -1933 mV for CaP-coated samples, was found significantly electropositive at -275 mV for Ga-doped ones. All samples were cytocompatible according to indirect tests. In vitro culture with Saos-2 cells led to changes in the surface compositions of the alloys. The numbers of cells attached to the Zn-doped (2.6 × 104 cells/cm2) and Ga-doped (6.3 × 104 cells/cm2) coatings were higher than that on the surface of the undoped coating (1.0 × 103 cells/cm2). Decreased corrosivity and enhanced cell affinity of the modified MgCa alloys (CaP coated and Zn and Ga doped, with Ga-doped ones having the greatest positive effect) make them novel and promising candidates as biodegradable metallic implant materials for the treatment of bone damages and other orthopedic applications.

The response of subcutaneous connective tissue to newly developed calcium phosphate-based root canal sealers.

AIM: To evaluate the histopathologic biocompatibility of two new calcium phosphate-based sealers (CPS-1 & CPS-2) with a commercially available calcium hydroxide-based sealer (Acroseal). METHODOLOGY: Polyethylene tubes were filled with freshly mixed sealers and implanted subcutaneously in the dorsal connective tissue of rats. Empty tubes were used as controls. Histopathological examinations were conducted at 7, 15, 30, 60 and 90 days after the implantation procedure. The presence of inflammation and predominant cell types were analysed statistically with Mann-Whitney U and Kruskal-Wallis non-parametric tests. Fibrous connective tissue thickness adjacent to each sample was recorded. Differences were tested for significance using anova and 'Duncan's' multiple comparison test (P < 0.05). RESULTS: CPS-1 sealer was associated with severe inflammation and remained an irritation throughout the 90-day implantation period; the tissue reaction pattern was stromal fibrosis. The control, CPS-2 and Acroseal sealers had similar patterns of irritation, which were more severe initially and diminished with time creating a thin fibrous capsule around the implant with a complete absence of inflammatory cells. There was no difference in tissue reaction between the control, CPS-1, CPS-2 and Acroseal groups amongst the first two observation periods (P > 0.05). However, there was a highly significant difference between the same groups at the last two observation periods (P < 0.01). Also, there were highly significant differences between the observation periods within all four groups at 7, 15, 30, 60 and 90 days (P < 0.01). CONCLUSION: CPS-1 sealer was not biocompatible. CPS-2 sealer and Acroseal had a favourable biocompatibility level based on the histological findings.

The Beneficial Role of Filipendula ulmaria Extract in Prevention of Prodepressant Effect and Cognitive Impairment Induced by Nanoparticles of Calcium Phosphates in Rats.

Mineral components of dental composites are used in many medical and dental applications, including preventive, restorative, and regenerative dentistry. To evaluate the behavioural alterations induced by nanosized particles of novel dental composites, by means of depressive level and cognitive functions, experimental groups of rats were chronically administered with nanosized hydroxyapatite (HA), tricalcium phosphate (TCP), and amorphous calcium phosphate (ACP) with or without simultaneous application of Filipendula ulmaria L. (FU) methanolic extract. The significant prodepressant action was observed in groups solely treated with HA and ACP. Besides, prolonged treatment with ACP also resulted in a significant decline in cognitive functions estimated in the novel object recognition test. The adverse impact of calcium phosphates on estimated behavioural functions was accompanied by increased oxidative damage and apoptotic markers in the prefrontal cortex, as well as diminished specific neurotrophin (BDNF) and gabaergic expression. The results of our investigation showed that simultaneous antioxidant supplementation with FU extract prevented calcium phosphate-induced behavioural disturbances, as well as prooxidative and apoptotic actions, with the simultaneous restoration of BDNF and GABA-A receptors in the prefrontal cortex. These findings suggest that FU may be useful in the prevention of prodepressant impact and cognitive decline as early as the manifestation of calcium phosphate-induced neurotoxicity.