Synthesis of Chitosan and Ferric-Ion (Fe3+)-Doped Brushite Mineral Cancellous Bone Scaffolds.

Biodegradable scaffolds are needed to repair bone defects. To promote the resorption of scaffolds, a large surface area is required to encourage neo-osteogenesis. Herein, we describe the synthesis and freeze-drying methodologies of ferric-ion (Fe3+) doped Dicalcium Phosphate Dihydrate mineral (DCPD), also known as brushite, which has been known to favour the in situ condition for osteogenesis. In this investigation, the role of chitosan during the synthesis of DCPD was explored to enhance the antimicrobial, scaffold pore distribution, and mechanical properties post freeze-drying. During the synthesis of DCPD, the calcium nitrate solution was hydrolysed with a predetermined stoichiometric concentration of ammonium phosphate. During the hydrolysis reaction, 10 (mol)% iron (Fe3+) nitrate (Fe(NO3)3) was incorporated, and the DCPD minerals were precipitated (Fe3+-DCPD). Chitosan stir-mixed with Fe3+-DCPD minerals was freeze-dried to create scaffolds. The structural, microstructural, and mechanical properties of freeze-dried materials were characterized.

[Preparation and in vivo osteogenesis of acellular dermal matrix/dicalcium phosphate composite scaffold for bone repair].

Objective: To investigate the physicochemical properties, osteogenic properties, and osteogenic ability in rabbit model of femoral condylar defect of acellular dermal matrix (ADM)/dicalcium phosphate (DCP) composite scaffold. Methods: ADM/DCP composite scaffolds were prepared by microfibril technique, and the acellular effect of ADM/DCP composite scaffolds was detected by DNA residue, fat content, and α-1，3-galactosyle (α-Gal) epitopes; the microstructure of scaffolds was characterized by field emission scanning electron microscopy and mercury porosimetry; X-ray diffraction was used to analyze the change of crystal form of scaffold; the solubility of scaffolds was used to detect the pH value and calcium ion content of the solution; the mineralization experiment in vitro was used to observe the surface mineralization. Twelve healthy male New Zealand white rabbits were selected to prepare the femoral condylar defect models, and the left and right defects were implanted with ADM/DCP composite scaffold (experimental group) and skeletal gold ® artificial bone repair material (control group), respectively. Gross observation was performed at 6 and 12 weeks after operation; Micro-CT was used to detect and quantitatively analyze the related indicators [bone volume (BV), bone volume/tissue volume (BV/TV), bone surface/bone volume (BS/BV), trabecular thickness (Tb.Th), trabecular number (Tb.N), trabecular separation (Tb.Sp), bone mineral density (BMD)], and HE staining and Masson staining were performed to observe the repair of bone defects and the maturation of bone matrix. Results: Gross observation showed that the ADM/DCP composite scaffold was a white spongy solid. Compared with ADM, ADM/DCP composite scaffolds showed a significant decrease in DNA residue, fat content, and α-Gal antigen content ( P<0.05). Field emission scanning electron microscopy showed that the ADM/DCP composite scaffold had a porous structure, and DCP particles were attached to the porcine dermal fibers. The porosity of the ADM/DCP composite scaffold was 76.32%±1.63% measured by mercury porosimetry. X-ray diffraction analysis showed that the crystalline phase of DCP in the ADM/DCP composite scaffolds remained intact. Mineralization results in vitro showed that the hydroxyapatite layer of ADM/DCP composite scaffolds was basically mature. The repair experiment of rabbit femoral condyle defect showed that the incision healed completely after operation without callus or osteophyte. Micro-CT showed that bone healing was complete and a large amount of new bone tissue was generated in the defect site of the two groups, and there was no difference in density between the defect site and the surrounding bone tissue, and the osteogenic properties of the two groups were equivalent. There was no significant difference in BV, BV/TV, BS/BV, Tb.Th, Tb.N, and BMD between the two groups ( P>0.05), except that the Tb.Sp in the experimental group was significantly higher than that in the control group ( P<0.05). At 6 and 12 weeks after operation, HE staining and Masson staining showed that the new bone and autogenous bone fused well in both groups, and the bone tissue tended to be mature. Conclusion: The ADM/DCP composite scaffold has good biocompatibility and osteogenic ability similar to the artificial bone material in repairing rabbit femoral condylar defects. It is a new scaffold material with potential in the field of bone repair.

Self-Healing Micro Arc Oxidation and Dicalcium Phosphate Dihydrate Double-Passivated Coating on Magnesium Membrane for Enhanced Bone Integration Repair.

Magnesium is a revolutionary biomaterial for orthopedic implants, owing to its eminent mechanical properties and biocompatibility. However, its uncontrolled degradation rate remains a severe challenge for its potential applications. In this study, we developed a self-healing micro arc oxidation (MAO) and dicalcium phosphate dihydrate (DCPD) double-passivated coating on a magnesium membrane (Mg-MAO/DCPD) and investigated its potential for bone-defect healing. The Mg-MAO/DCPD membrane possessed a feasible self-repairing ability and good cytocompatibility. In vitro degradation experiments showed that the Mg contents on the coating surface were 0.3, 3.8, 4.1, 6.1, and 7.9% when the degradation times were 0, 1, 2, 3, and 4 weeks, respectively, exhibiting available corrosion resistance. The slow and sustained release of Mg2+ during the degradation process activated extracellular matrix proteins for bone regeneration, accelerating osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs). The extract solutions of Mg-MAO/DCPD considerably promoted the activation of the Wnt and PI3K/AKT signaling pathways. Furthermore, the evaluation of the rat skull defect model manifested the outstanding bone-healing efficiency of the Mg-MAO/DCPD membrane. Taken together, the Mg-MAO/DCPD membrane demonstrates an optimized degradation rate and excellent bioactivity and is believed to have great application prospects in bone tissue engineering.

Development of calcium phosphate-chitosan composites with improved removal capacity toward tetracycline antibiotic: Adsorption and electrokinetic properties.

Two eco-friendly and highly efficient adsorbents, namely brushite-chitosan (DCPD-CS), and monetite-chitosan (DCPA-CS) composites were synthesized via a simple and low-cost method and used for tetracycline (TTC) removal. The removal behavior of TTC onto the composite particles was studied considering various parameters, including contact time, pollutant concentration, and pH. The maximum TTC adsorption capacity was 138.56 and 112.48 mg/g for the DCPD-CS and DCPA-CS, respectively. Increasing the pH to 11 significantly enhanced the adsorption capacity to 223.84 mg/g for DCPD-CS and 205.92 mg/g for DCPA-CS. The antibiotic adsorption process was well-fitted by the pseudo-second-order kinetic and Langmuir isotherm models. Electrostatic attractions, complexation, and hydrogen bonding are the main mechanisms governing the TTC removal process. Desorption tests demonstrated that the (NH4)2HPO4 solution was the most effective desorbing agent. The developed composites were more efficient than DCPD and DCPA reference samples and could be used as valuable adsorbents of TTC from contaminated wastewater.

Anti-adhesive effect of naturally obtained dicalcium phosphate dihydrate nanoparticles in the rat uterine wound model.

OBJECTIVE: In this study, we aimed to compare the anti-adhesive effects of contractubex and dicalcium phosphate dihydrate (DCPD) particles in rats treated with the uterine horn adhesion model. MATERIALS AND METHODS: Newly adult, 60 Wistar albino rats were used as experimental animals. The modified rat uterine horn adhesion model was used to induce intra-abdominal adhesion. Tumor necrosis factor (TNF)-α, interleukin (IL)-1, vascular endothelial growth factor (VEGF) and transforming growth factor (TGF)-ß1 were studied for biochemical and immunohistochemical examination. RESULTS: TNF-α decreased in each group, while it decreased more in G2 and G3 than in G1. IL-1ß decreased in each group, while it decreased the most in G3. TGF-ß1 and VEGF localization was less in the G2 compared to G1, the least TGF-ß1 and VEGF immunolocalization was detected in the G3 and G4. For both antibodies, the least localization among all groups belonged to G3. From day 7 to day 21, the highest TGF-ß1 immunolocalization was observed in G1, lesser localization in G2 and lowest in G3. CONCLUSION: DCPD nanoparticles show promise as a clinical antiadhesive agent and should be further evaluated in experimental animal models and human trials.

OBJETIVO: En este estudio, nuestro objetivo fue comparar los efectos antiadhesivos de las partículas de contractubex (CTX) y fosfato dicálcico dihidratado (DCPD) en ratas tratadas con el modelo de adhesión del cuerno uterino. MATERIALES Y MÉTODOS: Como animales de experimentación se utilizaron 60 ratas Wistar albinas, recién adultas. Se usó el modelo de adhesión del cuerno uterino de rata modificado para inducir la adhesión intraabdominal. Se estudiaron TNF-α, IL-1, VEGF y TGF-ß1 para examen bioquímico e inmunohistoquímico. RESULTADOS: el TNF-α disminuyó en cada grupo, mientras que disminuyó más en G2 y G3 que en G1. IL-1ß disminuyó en cada grupo, mientras que disminuyó más en G3. La localización de TGF-ß1 y VEGF fue menor en G2 en comparación con G1, la menor inmunolocalización de TGF-ß1 y VEGF se detectó en G3 y G4. Para ambos anticuerpos, la localización mínima entre todos los grupos pertenecía a G3. Desde el día 7 hasta el día 21, la mayor inmunolocalización de TGF-ß1 se observó en G1, menor localización en G2 y menor en G3. CONCLUSIÓN: las nanopartículas de DCPD se muestran prometedoras como agentes antiadhesivos clínicos y deben evaluarse más en modelos animales experimentales y ensayos en humanos.

Qualitative and Quantitative Profiling of Enamel Remineralizing Potential of Fluoride Varnishes Incorporating Bioactive Glass, Dicalcium Phosphate Dihydrate, and Modified MTA: A Raman Spectroscopic Study.

Aim: To evaluate and compare for remineralization potential of enamel at three different time intervals using commercially available MI Varnish and Duraflor Halo varnish [5% sodium fluoride (NaF) varnish] incorporating bioactive glass, dicalcium phosphate dihydrate (DCPD), and biomimetically modified mineral trioxide aggregate (MTA). Materials and methods: For the study, a buccolingual division into equal halves was done for 64 decoronated premolar crowns. Among these 64 crowns, we mounted 32 in acrylic discs to be used for microhardness and induced white spot lesions (WSLs) measuring 5 × 5 mm on the exposed enamel surface. These samples were divided into four groups of 24 each depending upon the varnish used. Varnish application was done, followed by immersion of these samples for 24 hours in artificial saliva, followed by peeling off of varnish. A pH cycling of 28 days was done after the varnish application before assessing the remineralization of the samples. Evaluation of all these samples was done at three different time intervals, baseline, after demineralization, and post remineralization for microhardness on the enamel surface. Raman spectroscopy was utilized for the measurement of phosphate (P) ion release. Sectioning of these samples to a thickness of 100 µm approximately was done to be viewed under a polarized light microscope. Results: Bioactive glass incorporated varnish showed the highest microhardness values, mineral content levels, and least depth of lesion posttreatment. Conclusion: All the experimental varnish showed significant remineralizing potential with the best potency seen with bioactive glass incorporated varnish. Clinical significance: The nonfluoride agents can be appropriately used in 2 wt% amount to augment the benefits of fluoride. How to cite this article: Hardikar AS, Gaonkar NN, Devendrappa SN, et al. Qualitative and Quantitative Profiling of Enamel Remineralizing Potential of Fluoride Varnishes incorporating Bioactive Glass, Dicalcium Phosphate Dihydrate, and modified MTA: A Raman Spectroscopic Study. Int J Clin Pediatr Dent 2023;16(2):363-370.

Effect of replacing dicalcium phosphate with mono-dicalcium phosphate to supplement phosphorus on laying performance, phosphorus-calcium metabolism and bone metabolism of aged laying hens.

The objective of this study was to evaluate the effect of replacing dicalcium phosphate (DCP) with mono-dicalcium phosphate (MDCP) to formulate low-phosphorus (P) diets on laying performance, egg quality, phosphorus-calcium metabolism, and bone metabolism of 69-78-week-old aged laying hens. Hy-Line Brown laying hens (n = 1,350, 69 weeks old) were randomly assigned to six treatments, each with five replicates of 45 hens. A corn-soybean meal-based diet was formulated to contain 0.12% non-phytate phosphorus (NPP), 3.81% calcium (Ca), and 1,470 FTU/kg phytase. The control group (CON) was supplemented with DCP inorganic phosphorus (Pi) at the NPP level of 0.20% (dietary NPP levels of 0.32%). Test groups (T1-T5) were supplemented with MDCP Pi at NPP levels of 0.07%, 0.11%, 0.15%, 0.18, and 0.20% (dietary NPP levels of 0.19, 0.23, 0.27, 0.30, and 0.32%, respectively). Calcium carbonate levels were adjusted to ensure all experimental diets contained the same Ca levels (3.81%). The feeding trial lasted 10 weeks, with hens increasing in age from 69 to 78 weeks. When supplemented with 1,470 FTU/kg phytase, extra DCP Pi or MDCP Pi did not affect (p > 0.05) laying performance (day laying rate, average egg weight, feed intake, feed-to-egg mass ratio, broken egg rate), egg quality (eggshell strength, albumen height, haugh units), or serum P, Ca, copper (Cu), iron (Fe), zinc (Zn), and manganese (Mn) levels. However, when laying hens were fed MDCP Pi (NPP levels of 0.07 to 0.20%), yolk color improved (p = 0.0148). The tibia breaking strength was significantly higher (p < 0.05) in the 0.18 and 0.20% NPP MDCP Pi groups than in the 0.20% NPP DCP Pi group. The breaking strength, Ca content, and P content of tibia in 0.11% and 0.15% NPP MDCP Pi hens were not significantly (p > 0.05) different from those in 0.20% NPP DCP Pi hens. Hens fed 0.07% NPP MDCP Pi had higher (p < 0.01) serum levels of osteoprotegerin (OPG), type-I collagen c-telopeptide (CTX-I), and tartrate-resistant acid phosphatase 5b (TRACP-5b) than those in all other groups. Serum levels of TRACP-5b and CTX-I in the 0.11% and 0.15% NPP MDCP Pi group were significantly lower than those in 0.18 and 0.20% NPP MDCP Pi groups and the 0.20% NPP DCP Pi group (p < 0.0001). Hens fed 0.07% and 0.11% NPP MDCP Pi had higher (p < 0.05) serum levels of parathyroid hormone (PTH) than those in all other groups. No differences were detected in serum calcitonin (CT), 1,25-dihydroxy-vitamin D3 (1,25-(OH)2D3), bone alkaline phosphatase (BAP), osteocalcin(OCN), and osteopontin (OPN) among all groups (p > 0.05). The expression of P transporters type IIa Na/Pi cotransporter (NaPi-IIa) in 0.11% and 0.15% NPP MDCP Pi hens were higher than those in 0.20% NPP MDCP Pi group and 0.20% NPP DCP Pi group (p < 0.05). The results indicated that both renal P reabsorption and bone resorption were involved in adapting to a low-P diet. In summary, when MDCP was used instead of DCP to supplement P, NPP levels could be reduced to 0.11% (dietary NPP level of 0.23%) without negative effects on laying performance and skeletal health of aged hens. In addition, MDCP was more beneficial than DCP for tibia quality. The results of the current study would provide references for the application of MDCP in low-P diets of aged laying hens.

Preliminary comparison of the remineralization and desensitization effects of a novel mineralized ion-releasing toothpaste and commercially available competitive products.

Background/purpose: Toothpaste plays an important role in brushing teeth to maintain oral hygiene and health. The purpose of this study was to develop a new toothpaste containing surface nanocrystal-rich dicalcium phosphate anhydrous (DCPA) powder and to investigate its effect on tooth samples. Materials and methods: The innovative toothpaste (REALCaP®/Group R) was compared with two commercial toothpastes (BioRepair®/Group B and Sensodyne®/Group S). Brushing cycle tests were performed on bovine tooth slices coated with individual toothpaste and a control group without toothpaste (Group C). Microhardness, roughness, surface structure observation, and X-ray diffraction (XRD) were performed on cycle days 4, 7, and 14 to analyze the impact of the toothpastes on tooth samples. Reults: Microhardness in the Group R was higher than that of the other groups regardless of the cycle days. Roughness in the Group R increased on days 4 and 7 but decreased on day 14. Roughness in the groups S and B increased with days. Microstructural observation revealed that most exposed dentinal tubules had been sealed in the Group R on day 14. Overlay thickness in the Group R was significantly higher than that in the groups S and B on days 4, 7, and 14. XRD analysis showed no hydroxyapatite (HA) peak in the Group S. The HA peak in the Group R was higher than that in the Group B on day 14. Conclusion: The innovative toothpaste has better properties than the commercially available products in terms of microhardness, roughness, and effectiveness in sealing dentinal tubules.

The effect of combination treatment of CO2-laser irradiation and tetracalcium phosphate/dicalcium phosphate anhydrate on dentinal tubules blockage: an in vitro study.

The aim of this study was the evaluation of the in vitro efficacy of a carbon dioxide (CO2) laser, a tetracalcium phosphate/dicalcium phosphate anhydrate (TP/DP) desensitizer and the combination of the desensitizer and additional CO2 laser irradiation as a treatment modality for cervical dentin hypersensitivity. A total of 48 dental specimens, prepared from extracted human premolars and molars, were divided into four groups: a control group, a TP/DP desensitizer paste group, a CO2 laser (10.600-nm wavelength) group, and a paste and laser group. The specimens were coated with nail varnish except in the marked area and were then immersed in 2% methylene blue dye for 1 h. The specimens were then washed, dried, and cut longitudinally. Thereafter, photos of 40 dentin specimens were taken and evaluated. The area of penetration was assessed and reported as percentage of the dentin surface area. Additionally eight dental specimens were examined with the aid of a scanning electron microscope and evaluated. Significant differences in the penetration depth were found for all experimental groups compared to the control group. The lowest penetration area was detected in the paste-laser group (16.5%), followed by the laser (23.7%), the paste (48.5%), and the control group (86.2%). The combined treatment of the CO2 laser and a TP/DP desensitizer was efficient in sealing the dentinal surface and could be a treatment option for cervical dentin hypersensitivity.

Hybrid Mineral/Organic Material Induces Bone Bridging and Bone Volume Augmentation in Rat Calvarial Critical Size Defects.

In craniofacial bone defects, the promotion of bone volume augmentation remains a challenge. Finding strategies for bone regeneration such as combining resorbable minerals with organic polymers would contribute to solving the bone volume roadblock. Here, dicalcium phosphate dihydrate, chitosan and hyaluronic acid were used to functionalize a bone-side collagen membrane. Despite an increase in the release of inflammatory mediators by human circulating monocytes, the in vivo implantation of the functionalized membrane allowed the repair of a critical-sized defect in a calvaria rat model with de novo bone exhibiting physiological matrix composition and structural organization. Microtomography, histological and Raman analysis combined with nanoindentation testing revealed an increase in bone volume in the presence of the functionalized membrane and the formation of woven bone after eight weeks of implantation; these data showed the potential of dicalcium phosphate dihydrate, chitosan and hyaluronic acid to induce an efficient repair of critical-sized bone defects and establish the importance of thorough multi-scale characterization in assessing biomaterial outcomes in animal models.

Dicalcium Phosphate Dihydrate Mineral Loaded Freeze-Dried Scaffolds for Potential Synthetic Bone Applications.

Dicalcium Phosphate Dihydrate (DCPD) mineral scaffolds alone do not possess the mechanical flexibility, ease of physicochemical properties' tuneability or suitable porosity required for regenerative bone scaffolds. Herein, we fabricated highly porous freeze-dried chitosan scaffolds embedded with different concentrations of Dicalcium Phosphate Dihydrate (DCPD) minerals, i.e., 0, 20, 30, 40 and 50 (wt)%. Increasing DCPD mineral concentration led to increased scaffold crystallinity, where the % crystallinity for CH, 20, 30, 40, and 50-DCPD scaffolds was determined to be 0.1, 20.6, 29.4, 38.8 and 69.9%, respectively. Reduction in scaffold pore size distributions was observed with increasing DCPD concentrations of 0 to 40 (wt)%; coalescence and close-ended pore formation were observed for 50-DCPD scaffolds. 50-DCPD scaffolds presented five times greater mechanical strength than the DCPD mineral-free scaffolds (CH). DCPD mineral enhanced cell proliferation for the 20, 30 and 40-DCPD scaffolds. 50-DCPD scaffolds presented reduced pore interconnectivity due to the coalescence of many pores in addition to the creation of closed-ended pores, which were found to hinder osteoblast cell proliferation.

Preparation and characterization of electrospun polycaprolactone/brushite scaffolds to promote osteogenic differentiation of mesenchymal stem cells.

Bone tissue engineering aims to develop effective strategies for repairing or replacing damaged bone tissue. In this study, composite scaffolds consisting of dicalcium phosphate dihydrate (DCDP, brushite) as a bone phase mineral precursor with different weight percentages (0%, 1%, 3%, 5%, and 10%) in combination with polycaprolactone (PCL) were fabricated by electrospinning technique. The morphology and mechanical behavior of scaffolds were characterized using scanning electron microscopy and tensile strength test, respectively. The bioactivity of scaffolds was assessed in simulated body fluid. Adhesion, viability, proliferation, and differentiation of mesenchymal stem cells derived from the human bone marrow on scaffolds were investigated using electron microscopy, MTT assay, live-dead assay, alizarin red staining, alkaline phosphatase activity and, gene expression analysis by real-time PCR. The results showed that the scaffold containing 3 wt. % of DCDP had the highest tensile strength (15.35 MPa). Furthermore, cells seeded on scaffolds showed over 80% viability after 1, 3, 7 days of incubation. Also, the results showed that the addition of DCDP to the PCL significantly increased the alkaline phosphatase activity. The osteocalcin gene expression in the composite scaffold showed a 6.1-fold increase compared to the pure PCL scaffold. It is concluded that electrospun PCL scaffolds containing DCDP with optimum concentration can be a proper candidate for bone tissue engineering applications.

3D gel-printed porous magnesium scaffold coated with dibasic calcium phosphate dihydrate for bone repair in vivo.

BACKGROUND: /Objective: The treatment of bone defect has always been a difficult problem in orthopedic clinic. The search for alternative biodegradable implants is a hot topic. The development of biodegradable magnesium scaffolds for the treatment of bone defects has long been a goal of the public. METHODS: In this study, we proposed a porous magnesium scaffold prepared by 3D gel printing and surface modification with an additional calcium phosphate coating and use of its strength, degradability and slow degradation rate in a bone graft substitute material. The porous magnesium granular scaffold was prepared by 3D gel printing technology and modified by DCPD (Dibasic Calcium Phosphate Dihydrate) coating. The biocompatibility, degradation rate, and osteogenic ability of the scaffold were evaluated in vitro and in vivo. RESULTS: The biocompatibility, in vivo degradation and bone defect healing response of the implants were investigated. Porous magnesium scaffolds were successfully prepared, and the strength of sintered scaffolds reached 5.38 â€‹MPa. The degradation rates of scaffolds were significantly reduced after coating with DCPD. The cell compatibility evaluation showed that DCPD-coated Mg scaffold was suitable for cell proliferation. In vivo biosafety monitoring showed that scaffold implantation did not cause an increase in Mg ion concentration in vivo, and no toxic damage was detected in the liver or kidney. Micro-CT and pathological results showed that a large amount of new bone was formed at 6 weeks. At 12 weeks, approximately 52% of the scaffold volume remained. At 24 weeks, osteogenesis, which was stimulated by some residual scaffold, still can be observed. In summary, this study suggests that 3D gel-printed DCPD-coated porous magnesium scaffolds have great potential as bone graft alternatives. CONCLUSION: In summary, this study suggests that 3D gel-printed DCPD-coated porous magnesium scaffolds have great potential as bone graft alternatives. THE TRANSLATIONAL POTENTIAL OF THIS ARTICLE: The translational potential of this article is to make use of the advantages of 3D gel printing technology with higher efficiency and lower cost compared with SLM and SLS technologies, and use pure magnesium powder as raw material to prepare degradable porous magnesium metal scaffolds, opening up a new technical route for the preparation of degradable porous magnesium scaffolds which are made for bone defect regeneration in the future.

Continuous direct compression: Development of an empirical predictive model and challenges regarding PAT implementation.

In this study, an empirical predictive model was developed based on the quantitative relationships between blend properties, critical quality attributes (CQA) and critical process parameters (CPP) related to blending and tableting. The blend uniformity and API concentration in the tablets were used to elucidate challenges related to the processability as well as the implementation of PAT tools. Thirty divergent ternary blends were evaluated on a continuous direct compression line (ConsiGma™ CDC-50). The trials showed a significant impact of the impeller configuration and impeller speed on the blending performance, whereas a limited impact of blend properties was observed. In contrast, blend properties played a significant role during compression, where changes in blend composition significantly altered the tablet quality. The observed correlations allowed to develop an empirical predictive model for the selection of process configurations based on the blend properties, reducing the number of trial runs needed to optimize a process and thus reducing development time and costs of new drug products. Furthermore, the trials elucidated several challenges related to blend properties that had a significant impact on PAT implementation and performance of the CDC-platform, highlighting the importance of further process development and optimization in order to solve the remaining challenges.

Fluoride and calcium release from peppermint-flavored fluoride varnish containing dicalcium-phosphate-dihydrate coated with xylitol.

Bakground: Fluoride varnish with high initial fluoride and calcium release can help patients with high-risk caries. Ample quantities of free fluoride and calcium ions in the oral cavity can enhance enamel remineralization. This study aimed to investigate the effect of dicalcium phosphate dihydrate coated with xylitol (DCPD-xylitol), in fluoride varnish, on the release of fluoride and calcium ions in the oral cavity. MATERIALS AND METHODS: DCPD powder with xylitol was synthesized by preparing a 60% xylitol solution and mixed it with DCPD solution. The mixture was stirred for 1 h at room temperature and dried at 80 °C for 18 h to reduce the water content. Then, the powder was used in the formulation of peppermint-flavored fluoride varnish as an active agent. The amounts of fluoride and calcium ion released in deionized water at 37 °C for 6 h were assessed with an ion-selective electrode. The cumulative fluoride and calcium ion release data were analyzed using one-way analysis of variance (ANOVA) and the post hoc Tukey test with α = 0.05. RESULTS: The results showed that the addition of DCPD coated with xylitol provided better bioavailability of the ions than DCPD without coating. Peppermint-flavored fluoride varnish (PFFV) with DCPD-xylitol 1% gave the highest fluoride ion release (296.90 mg/L) compared to the varnishes with other xylitol concentrations and the positive control. In contrast, PFFV DCPD-xylitol 5% afforded the highest calcium ion release at 111.20 mg/L. CONCLUSIONS: This study concluded that xylitol affects the bioavailability of free fluoride and calcium ions in varnishes. However, the efficacy of fluoride and calcium uptake in enamel and under different in vitro media conditions requires further investigation.

Green Hydrogels Composed of Sodium Mannuronate/Guluronate, Gelatin and Biointeractive Calcium Silicates/Dicalcium Phosphate Dihydrate Designed for Oral Bone Defects Regeneration.

Innovative green, eco-friendly, and biologically derived hydrogels for non-load bearing bone sites were conceived and produced. Natural polysaccharides (copolymers of sodium D-mannuronate and L-guluronate) with natural polypeptides (gelatin) and bioactive mineral fillers (calcium silicates CaSi and dicalcium phosphate dihydrate DCPD) were used to obtain eco-sustainable biomaterials for oral bone defects. Three PP-x:y formulations were prepared (PP-16:16, PP-33:22, and PP-31:31), where PP represents the polysaccharide/polypeptide matrix and x and y represent the weight % of CaSi and DCPD, respectively. Hydrogels were tested for their chemical-physical properties (calcium release and alkalizing activity in deionized water, porosity, solubility, water sorption, radiopacity), surface microchemistry and micromorphology, apatite nucleation in HBSS by ESEM-EDX, FT-Raman, and micro-Raman spectroscopies. The expression of vascular (CD31) and osteogenic (alkaline phosphatase ALP and osteocalcin OCN) markers by mesenchymal stem cells (MSCs) derived from human vascular walls, cultured in direct contact with hydrogels or with 10% of extracts was analysed. All mineral-filled hydrogels, in particular PP-31:31 and PP-33:22, released Calcium ions and alkalized the soaking water for three days. Calcium ion leakage was high at all the endpoints (3 h-28 d), while pH values were high at 3 h-3 d and then significantly decreased after seven days (p < 0.05). Porosity, solubility, and water sorption were higher for PP-31:31 (p < 0.05). The ESEM of fresh samples showed a compact structure with a few pores containing small mineral granules agglomerated in some areas (size 5-20 microns). PP-CTRL degraded after 1-2 weeks in HBSS. EDX spectroscopy revealed constitutional compounds and elements of the hydrogel (C, O, N, and S) and of the mineral powders (Ca, Si and P). After 28 days in HBSS, the mineral-filled hydrogels revealed a more porous structure, partially covered with a thicker mineral layer on PP-31:31. EDX analyses of the mineral coating showed Ca and P, and Raman revealed the presence of B-type carbonated apatite and calcite. MSCs cultured in contact with mineral-filled hydrogels revealed the expression of genes related to vascular (CD31) and osteogenic (mainly OCN) differentiation. Lower gene expression was found when cells were cultured with extracts added to the culture medium. The incorporation of biointeractive mineral powders in a green bio-derived algae-based matrix allowed to produce bioactive porous hydrogels able to release biologically relevant ions and create a suitable micro-environment for stem cells, resulting in interesting materials for bone regeneration and healing in oral bone defects.

Controlled synthesis of dicalcium phosphate dihydrate (DCPD) from metastable solutions: insights into pathogenic calcification.

Calcium phosphate (CaP) compounds may occur in the body as abnormal pathogenic phases in addition to their normal occurrence as bones and teeth. Dicalcium phosphate dihydrate (DCPD; CaPO4·2H2O), along with other significant CaP phases, have been observed in pathogenic calcifications such as dental calculi, kidney stones and urinary stones. While other studies have shown that polar amino acids can inhibit the growth of CaPs, these studies have mainly focused on hydroxyapatite (HAp; Ca10(PO4)6(OH)2) formation from highly supersaturated solutions, while their effects on DCPD nucleation and growth from metastable solutions have been less thoroughly explored. By further elucidating the mechanisms of DCPD formation and the influence of amino acids on those mechanisms, insights may be gained into ways that amino acids could be used in treatment and prevention of unwanted calcifications. The current study involved seeded growth of DCPD from metastable solutions at constant pH in the presence of neutral, acidic and phosphorylated amino acid side chains. As a comparison, solutions were also seeded with calcium pyrophosphate (CPP; Ca2P2O7), a known calcium phosphate inhibitor. The results show that polar amino acids inhibit DCPD growth; this likely occurs due to electrostatic interactions between amino acid side groups and charged DCPD surfaces. Phosphoserine had the greatest inhibitory ability of the amino acids tested, with an effect equal to that of CPP. Clustering of DCPD crystals giving rise to a "chrysanthemum-like" morphology was noted with glutamic acid. This study concludes that molecules containing an increased number of polar side groups will enhance the inhibition of DCPD seeded growth from metastable solutions.

Effects of Hinokitiol and Dicalcium Phosphate on the Osteoconduction and Antibacterial Activity of Gelatin-Hyaluronic Acid Crosslinked Hydrogel Membrane In Vitro.

Many hydrogel-based crosslinking membranes have been designed and tailored to meet the needs of different applications. The aim of this research is to design a bifunctional hydrogel membrane with antibacterial and osteoconducting properties to guide different tissues. The membrane uses gelatin and hyaluronic acid as the main structure, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride as the crosslinker, hinokitiol as the antibacterial agent, and dicalcium phosphate anhydrous (DCPA) micron particles for osteoconduction. Results show that the hydrogel membrane with added DCPA and impregnated hinokitiol has a fixation index higher than 88%. When only a small amount of DCPA is added, the tensile strength does not decrease significantly. The tensile strength decreases considerably when a large amount of modified DCPA is added. The stress-strain curve shows that the presence of a large amount of hinokitiol in hydrogel membranes results in considerably improved deformation and toughness properties. Each group impregnated with hinokitiol exhibits obvious antibacterial capabilities. Furthermore, the addition of DCPA and impregnation with hinokitiol does not exert cytotoxicity on cells in vitro, indicating that the designed amount of DCPA and hinokitiol in this study is appropriate. After a 14-day cell culture, the hydrogel membrane still maintains a good shape because the cells adhere and proliferate well, thus delaying degradation. In addition, the hydrogel containing a small amount of DCPA has the best cell mineralization effect. The developed hydrogel has a certain degree of flexibility, degradability, and bifunctionality and is superficial. It can be used in guided tissue regeneration in clinical surgery.

Transformation of acellular dermis matrix with dicalcium phosphate into 3D porous scaffold for bone regeneration.

Animal derived biomaterials have attracted much attentions in treating large size bone defect due to their excellent biocompatibility and potent bioactivities offered by the biomacromolecules and growth factors contained in these materials. Dermis-derived matrix (ADM) has been used as skin grafts and wound dressings for decades, however its application in bone tissue engineering has been largely limited as ADM possesses a dense structure which does not support bone tissue ingrowth. Recently, we have successfully fabricated porous scaffold structure using an ADM with the aid of micronization technique. When integrated with inorganic components such as calcium phosphate, ADM could be transformed to bone graft substitutes with desirable osteogenic properties. While purified and chemically cross-linked collagen has lost its natural structure, our ADM successfully preserved natural tropocollagen structure, as well as other bioactive components. A composite scaffold was fabricated by incorporating dicalcium phosphate (DCP) microparticles into ADM microfibers and freeze-dried to form a highly porous structure. Unlike conventional ADM materials, this scaffold possesses high porosity with interconnected pores. More importantly, our evaluation data demonstrated that it performed much more effective in treating critical bone defects in comparison with best commercial product on the market. In a head-to-head comparison with a commercial bone graft material Bongold®, the ADM/DCP scaffold showed superior osteogenic capacity by filling the defect with well-organized new bone tissue in a rabbit radius segmental defect model. Put together, our results exhibited a novel bone graft substitute was developed by circumventing processing barriers associated with natural ADM, which offers another novel bone graft substitute for bone regeneration.

Effects of pullulan on the biomechanical and anti-collapse properties of dicalcium phosphate dihydrate bone cement.

In this work, a modified dicalcium phosphate dihydrate (DCPD) bone cement with unique biodegradable ability in a calcium phosphate cement system was prepared by the hydration reaction of monocalcium phosphate monohydrate and calcium oxide and integration with pullulan (Pul), a non-toxic, biocompatible, viscous, and water-soluble polysaccharide that has been successfully used to improve defects in DCPD bone cement, especially its rapid solidification, fragile mechanical properties, and easy collapse. The effect of different contents of Pul on the structure and properties of DCPD were also studied in detail. The modified cement was characterised by X-ray diffraction, Fourier-transform infrared spectroscopy, scanning electron microscopy, ultraviolet-visible absorption, X-ray photoelectron spectroscopy analysis, and rheological property measurements. The results indicated that Pul promoted the hydration formation of DCPD, and interface bonding occurred between Pul and DCPD. With increasing content of Pul, the setting time of the DCPD bone cement increased from 2.6 min to 42.3 min, the compressive strength increased from 0 MPa to 20.4 MPa, and the anti-collapse ability also improved owing to the strong interface bonding, implying that the DCPD bone cement improved by Pul has better potential for application in the field of non-loading bone regenerative medicine compared to unmodified DCPD bone cement.