Bioactive materials in the traumatology of small pets

The effect of the biocomposition material was studied on the basis of synthetic hydroxyapatite, collagen and antibiotic for reparative histogenesis of bone tissue during shoulder bone fractures and forearm in dogs and implants for the tissue regeneration of the new generation Xenograft Collagen.It has been established that remodeling of the primary bone corn into the secondary in the region of the fracture in the dogs studied was diverse. During the experiment, it was revealed that a month after the operation on radiographs in the control group of dogs, there was a defect in which the formation of a weak periosal bone corn was observed. Endostal bone corn has not been detected. The most pronounced process proceeded by dogs in experienced groups. In the first experimental defect, a collapan was filled with a drug, and in the second used the implant in the form of the xenograft collagen granules. In animals of the first experienced group, the primary bone-cartilage corn had a more mature and developed character, intermediary and periosal corns were formed.

Bonding Reactions of Dental Self-Adhesive Cements with Synthetic Hydroxyapatite as a Function of the Polymerization Protocol

Objectives. This study evaluated the influence of the cement composition and different polymerization protocols on the bonding chemical interaction of self-adhesive cements with synthetic hydroxyapatite. Materials and Methods. Two commercial self-adhesive resin cements (RelyX U200 and Maxcem Elite) were selected, manipulated, mixed with hydroxyapatite dry powder (HAp), dispensed into molds, and distributed into three groups according to polymerization protocols: immediate photoactivation (IP); delayed photoactivation, 10 min self-curing and light-curing (DP); and chemical activation (CA, no light exposure). The detailed chemical information, at atomic scale, on the surface and deeper into the bulk of self-adhesive cement/hydroxyapatite mixtures was evaluated with X-ray photoelectron spectroscopy (XPS). Results. Chemical elements were detected in both cements, such as Na, O, Ca, C, P, and Si. Other elements were detected in minor concentrations. RelyX U200 exhibited the most intense formation of calcium salts products when the cement/HAp mixtures were photoactivated (immediate or delayed). RelyX U200/HAp mixture under delayed photoactivation (DP) also exhibited higher binding energy between calcium moieties of the HAp and methacrylates in the cement. A higher energy difference in the interaction of HAp with the cement comparing the bulk and surface areas was observed when RelyX U200 underwent the delayed photoactivation protocol. Maxcem Elite exhibited an increased chemical reactivity when either chemically activated or immediately photoactivated and a higher binding energy of the carboxyl groups bonded to the calcium of HAp when chemically activated. Conclusions. The interaction of cements with hydroxyapatite is chemical in nature and leads to the formation of calcium salts, which may favor better integrity and longevity of adhesive restorations. The polymerization protocol affects the chemical interaction in mixtures of self-adhesive cements and hydroxyapatite, influencing the formation of these salts and the establishment of intermolecular interactions between the HAp and the cements.

Repair of Critical Size Bone Defects Using Synthetic Hydroxyapatite or Xenograft with or without the Bone Marrow Mononuclear Fraction: A Histomorphometric and Immunohistochemical Study in Rat Calvaria

Bone defects are a challenging clinical situation, and the development of hydroxyapatite-based biomaterials is a prolific research field that, in addition, can be joined by stem cells and growth factors in order to deal with the problem. This study compares the use of synthetic hydroxyapatite and xenograft, used pure or enriched with bone marrow mononuclear fraction for the regeneration of critical size bone defects in rat calvaria through histomorphometric (Masson’s staining) and immunohistochemical (anti-VEGF, anti-osteopontin) analysis. Forty young adult male rats were divided into five groups (n = 8). Animals were submitted to critical size bone defects (Ø = 8 mm) in the temporoparietal region. In the control group, there was no biomaterial placement in the critical bone defects; in group 1, it was filled with synthetic hydroxyapatite; in group 2, it was filled with xenograft; in group 3, it was filled with synthetic hydroxyapatite, enriched with bone marrow mononuclear fraction (BMMF), and in group 4 it was filled with xenograft, enriched with BMMF. After eight weeks, all groups were euthanized, and histological section images were captured and analyzed. Data analysis showed that in groups 1, 2, 3 and 4 (received biomaterials and biomaterials plus BMMF), a significant enhancement in new bone matrix formation was observed in relation to the control group. However, BMMF-enriched groups did not differ from hydroxyapatite-based biomaterials-only groups. Therefore, in this experimental model, BMMF did not enhance hydroxyapatite-based biomaterials’ potential to induce bone matrix and related mediators.

POS0366 PRO-INFLAMMATORY EFFECTS OF HUMAN APATITE CRYSTALS EXTRACTED FROM PATIENTS SUFFERING FROM CALCIFIC TENDINOPATHY

Background:Calcific tendonitis of the rotator cuff is due to carbonated apatite deposits in the shoulder tendons. During the evolution of the disease, an acute inflammatory episode may occur leading to the disappearance of the calcification. Although hydroxyapatite crystals-induced inflammation has been previously studied with synthetic crystals, no data are available with calcifications extracted from patients suffering from calcific tendinopathy. The objective of the study was to explore the inflammatory properties of human calcifications and the pathways involved.Objectives:The objective of the study was to explore the inflammatory properties of human calcifications and the pathways involved.Methods:Human calcifications were obtained from patients treated for their shoulder pain related to a calcific tendinopathy of the rotator cuff. Calcifications were extracted by ultrasound-guided lavage and aspiration as previously described [1]. Human calcifications and synthetic hydroxyapatite (sHA) were used in vitro to stimulate human monocytes and macrophages, the human myeloid cell line THP-1 and human tenocytes. The release of IL-1β, IL-6 and IL-8 by cells was quantified by ELISA. Gene expression of pro- and anti-inflammatory cytokines was evaluated by quantitative PCR. NF-kB activation and NLRP3 involvement was assessed in THP-1 cells using a NF-kB inhibitor and a Caspase 1 inhibitor. The inflammatory properties were then assessed in vivo using a mouse air pouch model. The membrane thickness and infiltrate were assessed 6 and 24 hours after the injection of human calcifications or synthetic hydroxyapatite using hematoxylin and eosin staining. Macrophages, neutrophils and lymphocytes infiltrates were assessed by immunohistochemistry. Total RNA was extracted from the membranes and expression of IL-1β, IL-6 and TNFβ was quantified by PCR.Results:Human calcifications were able to induce a significant release of IL-1β when incubated with monocytes, macrophages and THP-1 only if they were first primed with LPS (lipopolysaccharide) for monocytes and macrophages or PMA (Phorbol 12-myristate 13-acetate) for THP-1. No IL-1β was detected in tenocytes’ supernatants. Stimulation of THP-1 by human calcifications led to similar levels of IL-1β when compared to synthetic hydroxyapatite although these levels were significantly inferior in monocytes and macrophages. IL-6 and IL-8 levels were not increased in the supernatants after crystal stimulation. Patient’s crystals enhanced mRNA expression of pro-IL-1β, as well as IL-18, NF-kB and TGFβ when IL-6 and TNFα expression were not. IL-1β production was reduced by the inhibition NF-kB as well as Caspase 1 indicating the role of NLRP3 inflammasome. In vivo, injection of human calcifications or synthetic hydroxyapatite in air pouch led to significant increase in membrane thickness with an infiltrate mainly composed of macrophages. Significant overexpression of IL-1β was only observed in the synthetic hydroxyapatite group.Conclusion:As synthetic hydroxyapatite, human calcifications were able to induce an inflammatory response resulting in the production of IL-1β after NF-kB activation and through NLRP3 inflammasome. In some experiments, IL-1β induction was lower with human calcifications compared to synthetic apatite. Differences in size, shape and protein content may explain this observation.References:[1]Darrieutort-Laffite C, Arnolfo P, Garraud T, Adrait A, Couté Y, Louarn G, et al. Rotator Cuff Tenocytes Differentiate into Hypertrophic Chondrocyte-Like Cells to Produce Calcium Deposits in an Alkaline Phosphatase-Dependent Manner. J Clin Med. 2019 Sep 26;8(10):1544. doi: 10.3390/jcm8101544.Acknowledgements:Fondation Arthritis, Recherche et Rhumatismes and French Society for Rheumatology for their financial supportDisclosure of Interests:None declared

Pro-inflammatory effects of human apatite crystals extracted from patients suffering from calcific tendinopathy

Background Calcific tendonitis of the rotator cuff is due to carbonated apatite deposits in the shoulder tendons. During the evolution of the disease, an acute inflammatory episode may occur leading to the disappearance of the calcification. Although hydroxyapatite crystal-induced inflammation has been previously studied with synthetic crystals, no data are available with calcifications extracted from patients suffering from calcific tendinopathy. The objective of the study was to explore the inflammatory properties of human calcifications and the pathways involved. Methods Human calcifications and synthetic hydroxyapatite were used in vitro to stimulate human monocytes and macrophages, the human myeloid cell line THP-1, and human tenocytes. The release of IL-1β, IL-6, and IL-8 by cells was quantified by ELISA. The gene expression of pro- and anti-inflammatory cytokines was evaluated by quantitative PCR. NF-kB activation and NLRP3 involvement were assessed in THP-1 cells using a NF-kB inhibitor and a caspase-1 inhibitor. The inflammatory properties were then assessed in vivo using a mouse air pouch model. Results Human calcifications were able to induce a significant release of IL-1β when incubated with monocytes, macrophages, and THP-1 only if they were first primed with LPS (monocytes and macrophages) or PMA (THP-1). Stimulation of THP-1 by human calcifications led to similar levels of IL-1β when compared to synthetic hydroxyapatite although these levels were significantly inferior in monocytes and macrophages. The patient’s crystals enhanced mRNA expression of pro-IL-1β, as well as IL-18, NF-kB, and TGFβ when IL-6 and TNFα expression were not. IL-1β production was reduced by the inhibition of caspase-1 indicating the role of NLRP3 inflammasome. In vivo, injection of human calcifications or synthetic hydroxyapatite in the air pouch led to a significant increase in membrane thickness although significant overexpression of IL-1β was only observed for synthetic hydroxyapatite. Conclusions As synthetic hydroxyapatite, human calcifications were able to induce an inflammatory response resulting in the production of IL-1β after NF-kB activation and through NLRP3 inflammasome. In some experiments, IL-1β induction was lower with human calcifications compared to synthetic apatite. Differences in size, shape, and protein content may explain this observation.

Pro-inflammatory effects of human apatite crystals extracted from patients suffering from calcific tendinopathy.

Background: Calcific tendonitis of the rotator cuff is due to carbonated apatite deposits in the shoulder tendons. During the evolution of the disease, an acute inflammatory episode may occur leading to the disappearance of the calcification. Although hydroxyapatite crystals-induced inflammation has been previously studied with synthetic crystals, no data are available with calcifications extracted from patients suffering from calcific tendinopathy. The objective of the study was to explore the inflammatory properties of human calcifications and the pathways involved. Methods: Human calcifications and synthetic hydroxyapatite were used in vitro to stimulate human monocytes and macrophages, the human myeloid cell line THP-1 and human tenocytes. The release of IL-1β, IL-6 and IL-8 by cells was quantified by ELISA. Gene expression of pro- and anti-inflammatory cytokines was evaluated by quantitative PCR. NF-kB activation and NLRP3 involvement was assessed in THP-1 cells using a NF-kB inhibitor and a Caspase 1 inhibitor. The inflammatory properties were then assessed in vivo using a mouse air pouch model. Results: Human calcifications were able to induce a significant release of IL-1β when incubated with monocytes, macrophages and THP-1 only if they were first primed with LPS (monocytes and macrophages) or PMA (THP-1). Stimulation of THP-1 by human calcifications led to similar levels of IL- 1β when compared to synthetic hydroxyapatite although these levels were significantly inferior in monocytes and macrophages. Patient’s crystals enhanced mRNA expression of pro-IL-1β, as well as IL-18, NF-kB and TGFβ when IL-6 and TNFα expression were not. IL-1β production was reduced by the inhibition of Caspase 1 indicating the role of NLRP3 inflammasome. In vivo, injection of human calcifications or synthetic hydroxyapatite in air pouch led to significant increase in membrane thickness although significant overexpression of IL-1β was only observed for synthetic hydroxyapatite. Conclusions: As synthetic hydroxyapatite, human calcifications were able to induce an inflammatory response resulting in the production of IL-1β after NF-kB activation and through NLRP3 inflammasome. In some experiments, IL-1β induction was lower with human calcifications compared to synthetic apatite. Differences in size, shape and protein content may explain this observation.

Sorption interactions in system «hydroxyapatite—aqueous solution of CuSO4»

Regularities of adsorption of copper (II) ions by synthetic hydroxyapatite have been examined. It is shown that sorption of metal ions occurs mainly in the mixed diffusion region, the chemical interaction described by the pseudo-second order kinetic equation has a significant effect on the process speed. It was found out that that the adsorption isotherm was described by the Langmuir monomolecular adsorption equation in the best way. Interaction specificity of hydroxyapatite with water was studied by NMR relaxation spectroscopy method