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Mandibular continuity defects stem from conditions such as malignancies, trauma, cysts, osteomyelitis and osteoradionecrosis, presenting significant challenges. If mandibular reconstruction fails, it can result in facial collapse, causing significant aesthetic and functional concerns for the patient. In the present study we developed a bio-adhesive Bone Cement (BC) enriched with lyophilised PRF and gelatin to enhance bone repair and induce regeneration. The developed BC consisted of a mixture of Tetracalcium Phosphate (TTCP) and O-Phospho-l-serine (OPLS) in addition to lyophilised Platelet Rich Fibrin (PRF) for sustained growth factor release and gelatin (GE) for improved cement resorption. It is primarily designed for in-situ application, conforming to the shape and size of the defect for effective bone repair and regeneration. The study evaluated four groups: (i) BC (control), (ii) BC-GE (control), (iii) BC-PRF, and (iv) BC-GE-PRF. All the four groups were characterised using FTIR, SEM and XRD. The mechanical studies of the prepared beads exhibited a significant increase in the compressive strength of the PRF loaded bone cement composites. In vitro degradation study of the beads over a 60-day period revealed a significantly higher percentage of bone cement resorption in the gelatin-incorporated groups, BC-GE (44 ± 0.5 %), and BC-GE-PRF (45 ± 2 %). The assessment of growth factor release (TGF-ß and VEGF) using ELISA revealed a prolonged and sustained release of both growth factors over a 28-day period. In vitro studies were performed on human Dental Follicle Stem Cells (DFSCs) to assess cell attachment, proliferation, mineralisation and osteogenic differentiation. These studies clearly depicted that BC-PRF and BC-GE-PRF showed significantly greater proliferation of DFSCs. Furthermore, BC-PRF and BC-GE-PRF samples exhibited notably elevated expression of Runx2 and OPN (osteogenic markers), as well as a higher intensity of alizarin red stain (mineralisation). Therefore, it was concluded that PRF incorporated bioadhesive bone cement composites greatly enhance the cell attachment, proliferation, mineralisation and osteogenic differentiation of the DFSCs. Thus, the PRF and gelatin incorporated bone cement composites is expected to facilitate effective and faster bone regeneration and healing in a wide range of dental and maxillofacial defects.

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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.

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In this research, a photocurable composite based on tetracalcuim phosphate ceramic and, hydroxyethyl methacrylate-modified poly(acrylic-maleic acid) was developed and studied as a potential drug delivery system for bone defects. Different concentrations (5, 10 and 20 wt. %) of a non-steroidal anti-inflammatory drug, Indomethacin, were loaded on to the composite and its release behavior was investigated in phosphate buffered solution during 504 h. The obtained release data were fitted by both power law (Peppas) and Weibull equations. The composites were also characterized after different soaking periods using X-ray diffractometry (XRD), scanning electron microscopy (SEM) and Fourier transforming infrared spectroscopy. The results of XRD and SEM analyses revealed the formation of nanosized needle/flake-like apatite crystals on the composites surfaces; however, better apatite formation was observed for the composites loaded with higher amounts of Indomethacin. The morphological observations and quantitative estimations revealed that the loaded composites were gradually degraded in the phosphate-buffered saline. Moreover, a controlled release of Indomethacin was found from the composites in which a higher drug concentration led to a more drug level as well as sustained release profile. In drug release modeling, better regression coefficient was obtained from the Weibull equation, compared to the power law, meaning that the Weibull equation suggests a better description of the indomethacin release from the composites during the whole period of the test. In conclusion, the photocurable composite with apatite formation ability can be successfully used for the controlled release of indomethacin as an anti-inflammatory drug in bone defects.

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Background/purpose: Bioceramic tetracalcium phosphate (TTCP) is used as a surface modifier on the implant surface and the clinical studies on this surface modification are still limited. The objective of this clinical study was to investigate short-term implant stability of titanium implant surfaces being modified through sandblasting and acid etching (SLA), followed by TTCP sintered bioceramic anchoring. Materials and methods: A total of 20 patients who had single tooth space were included in this study. Surface modification by SLA plus with TTCP on Ti implants with a diameter of 4.0 mm and lengths of 10 and 11.5 mm were placed. Implant stability quotient (ISQ) value was measured immediately (ISQ0) and one month (ISQ1), two months (ISQ2), three months (ISQ3), and four months (ISQ4) after implantation. Subgroup analysis was defined to location (maxilla, mandible) and bone density (soft or hard bone). Statistical analysis was performed using Friedman test and Mann-Whitney U test. Results: The mean ISQ values with standard deviation at the different time points of ISQ0 to ISQ4 were 60.03 ± 14.12, 53.48 ± 15.24, 58.91 ± 14.43, 63.14 ± 12.22, and 63.50 ± 13.61, respectively. The results showed significant differences between the ISQ1 and ISQ3 groups and between the ISQ1 and ISQ4 groups. On the other hand, there was no statistical differences between the maxilla and mandible as well as between soft and hard bone types in all implant groups. Conclusion: TTCP/titanium implant showed favorable stability in short-term ISQ values over 4 months. The locations and bone types demonstrated no effect on implant stability.

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Polymeric and tetracalcium phosphate (TTCP)-containing polymeric scaffolds were fabricated using a freeze-drying technique, with a homogenous solution of hydroxyethyl cellulose (HEC)/hyaluronic acid (HA)/gelatin (G) or suspension of 15 or 20% TTCP) particles in HEC/HA/G solution. The morphology, phase composition, chemical bands, and swelling behavior of the scaffold were determined. In vitro fibroblast cell viability and migration potential of the scaffolds were determined by MTT, live/dead staining, and scratch assay for wound healing. The in vivo chick embryo angiogenesis test was also carried out. Finally, the initial antibacterial activity of the scaffolds was determined using Staphylococcus aureus. The scaffolds exhibited an enormous porous structure in which the size of pores increased by the presence of TTCP particles. While the polymeric scaffold was amorphous, the formation of low crystalline hydroxyapatite phase and the initial TTCP particles was determined in the composition of TTCP-added scaffolds. TTCP increased swelling behavior of the polymeric scaffold in PBS. The results demonstrated that the amount of TTCP was a crucial factor in cell life. A high concentration of TTCP could restrict cell viability, although all the scaffolds were nontoxic. The scratch assessments determined better cell migration and wound closure in treating with TTCP-containing scaffolds so that after 24 h, a wound closure of 100% was observed. Furthermore, TTCP-incorporated scaffolds significantly improved the angiogenesis, in the chick embryo test. The presence of TTCP had a significant effect on reducing the bacterial activity and 20% TTCP-containing scaffold exhibited better antibacterial activity than the others.

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Tetracalcium phosphate (TTCP), a potential biological scaffold material, has attracted increasing interest for bone regeneration applications due to its good biodegradability and biocompatibility. In this research, three-dimensional porous TTCP scaffolds were manufactured via selective laser sintering (SLS), and an in-depth and meticulous study on the influence of laser power on the microstructure and mechanical properties of TTCP scaffolds was performed. The results showed that the TTCP particles fused together and formed a solid object due to the decrease in the number of micro-pores in the scaffold as the laser power increased from 6 W to 9 W. The maximum compressive strength that the scaffold could withstand and the strength of the fracture toughness were 11.87 ± 0.64 MPa and 1.12 ± 0.1 MPa·m1/2, respectively. When the laser power increased from 9 W to 10 W, the TTCP grains grew abnormally, resulting in diminished mechanical properties. The bioactivity tests showed that the surfaces of the scaffolds were entirely covered by bone-like apatite layers after soaking in simulated body fluid (SBF) for three days, indicating that the scaffolds exhibit excellent bioactivity. Moreover, cell experiments showed that the TTCP scaffolds had good biocompatibility. This study indicated that SLS-fabricated TTCP scaffolds may be a promising candidate for bone regeneration applications.

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We previously described the gelation mechanism of calcium polyphosphate (CPP) in the presence of water. In this study, we developed novel and injectable poly-dicalcium phosphate dihydrate (P-DCPD) forming cement by the reaction of acidic CPP gel with alkali tetracalcium phosphate (TTCP). The setting reaction mechanism of P-DCPD is due to the intermolecular interaction between CPP gel and TTCP that was supported by XRD, AFM, Raman spectra analysis and SEM. The setting mechanism of P-DCPD is completely different from the classical calcium phosphate cement (CPC) that achieves crystallization by monophosphates reaction. P-DCPD represents a new type of poly-CPCs with significant advantages, including strong mechanical strength, excellent cohesion and easy of handling. More extensive experiments are currently underway to further evaluate the performance of P-DCPD cements, including biocompatibility, degradation behavior and bone defect hearing efficacy, among others.

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OBJECTIVES: To investigate the tubule occlusion and remineralization potential of a novel toothpaste with active tetracalcium phosphate/monetite mixtures under de/remineralization cycling. METHODS: Dentin de/remineralization cycling protocol consisted of demineralization in 1% citric acid at pH 4.6 with following remineralization with toothpastes and soaking in artificial saliva. Effectiveness of toothpastes to promote remineralization was evaluated by measurement of microhardness recovery, analysis of surface roughness, thickness of coating and scanning electron microscopy. RESULTS: The novel tetracalcium phosphate/monetite dentifrice had comparable remineralization potential as commercial calcium silicate/phosphate (SENSODYNE®) and magnesium aluminum silicate (Colgate®) toothpastes and significantly higher than control saliva (p<0.02). Surface roughness was significantly lower after treatment with prepared and SENSODYNE® dentifirice (p<0.05). The coatings on dentin surfaces was significantly thicker after applying toothpastes as compared to negative control (p<0.001). CONCLUSIONS: The new fluoride toothpaste formulation with bioactive tetracalcium phosphate/monetite calcium phosphate mixture effectively occluded dentin tubules and showed good dentin remineralization potential under de/remineralization cycling. It could replace professional powder preparation based on this mixture. It was demonstrated that prepared dentifrice had comparable properties with commercial fluoride calcium silicate/phosphate or magnesium aluminum silicate dentifrices.

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Classic bone wax is associated with drawbacks such as the risk of infection, inflammation and hindered osteogenesis. Here, we developed a novel self-setting bone wax on the basis of hydrophilic poly(ethylene glycol) (PEG) and hydroxyapatite (HA) forming calcium phosphate cement (CPC), to overcome the problems that are linked to the use of conventional beeswax systems. Amounts of up to 10 wt.% of pregelatinized starch were additionally supplemented as hemostatic agent. After exposure to a humid environment, the PEG phase dissolved and was exchanged by penetrating water that interacted with the HA precursor (tetracalcium phosphate (TTCP)/monetite) to form highly porous, nanocrystalline HA via a dissolution/precipitation reaction. Simultaneously, pregelatinized starch could gel and supply the bone wax with liquid sealing features. The novel bone wax formulation was found to be cohesive, malleable and after hardening under aqueous conditions, it had a mechanical performance (∼2.5 MPa compressive strength) that is comparable to that of cancellous bone. It withstood systolic blood pressure conditions for several days and showed antibacterial properties for almost one week, even though 60% of the incorporated drug vancomycin hydrochloride was already released after 8h of deposition by diffusion controlled processes. STATEMENT OF SIGNIFICANCE: The study investigated the development of alternative bone waxes on the basis of a hydroxyapatite (HA) forming calcium phosphate cement (CPC) system. Conventional bone waxes are composed of non-biodegradable beeswax/vaseline mixtures that are often linked to infection, inflammation and hindered osteogenesis. We combined the usage of bioresorbable polymers, the supplementation with hemostatic agents and the incorporation of a mineral component to overcome those drawbacks. Self-setting CPC precursors (tetracalcium phosphate (TTCP), monetite) were embedded in a resorbable matrix of poly(ethylene glycol) (PEG) and supplemented with pregelatinized starch. This formulation was found to be malleable and cohesive underwater. While immersion in an aqueous environment, CPC precursors formed highly porous, nanocrystalline HA via dissolution/precipitation reaction as water penetrated the novel wax formulation and PEG molecules simultaneously dissolved. The bone wax further withstood blood pressure conditions. After hardening, mechanical performance was comparable to that of cancellous bone and we also successfully provided the bone wax with antibacterial properties. In our opinion, the described bone wax formulation outmatches conventional bone waxes, as it circumvents the detriments being associated with the term "bone wax". Our wax has a novel composition and would broaden the application of CPC and besides, the general interest in bone waxes will increase, as they were long considered as a "first-line treatment" to avoid.

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The purpose of this study was to examine the in vitro cytocompatibility of a novel injectable multiphasic bone substitute (MBS) based on polysaccharide gel-coated OSPROLIFE(®) hydroxyapatite (HA)/tetracalcium phosphate (TTCP) granules combined with bone marrow concentrate (BMC). Polysaccharide gel-coated granules loaded in syringe were combined with BMC diluted in ionic crosslinking solution. The product was then maintained in culture to investigate the cytocompatibility, distribution, and osteogenic differentiation function of cells contained in the BMC. The in vitro cytocompatibility was assessed after 0, 24, and 96 h from the injectable MBS preparation using the LIVE/DEAD(®) staining kit. The results highlighted that cells remained viable after combination with the polysaccharide gel-coated granules; also, viability was maintained over time. The distribution of the cells in the product, observed using confocal microscopy, showed viable cells immersed in the polysaccharide gel formed between the granules after ionic crosslinking. The mesenchymal stromal cells (MSC) contained in the injectable MBS, the basic elements for bone tissue regeneration, were able to differentiate toward osteoblasts, producing an osteogenic matrix as evidenced by alizarin red-s (AR-S) staining. In conclusion, we found that the injectable MBS may have the potential to be used as a bone substitute by applying a "one-step" procedure in bone tissue engineering applications. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 894-902, 2016.

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Well-designed implants are used not only to modify the geometry of the implant but also to change the chemical properties of its surfaces. The present study aims to assess the biofunctional effects of tetracalcium phosphate (TTCP) particles as a physical anchor on the implant surface derived through sandblasting. The characteristics of the surface, cell viability, and alkaline phosphatase (ALP) activity toward osteoprogenitor cells (D1) were obtained. D1 cells were cultured on a plain surface that underwent sandblasting and acid etching (SLA) (control SLA group) and on different SLA surfaces with different anchoring TTCP rates (new test groups, M and H). The mean anchoring rates were 57% (M) and 74% (H), and the anchored thickness was estimated to range from 12.6µm to 18.3µm. Compared with the control SLA surface on Ti substrate, the new test groups with different TTCP anchoring rates (M and H) failed to improve cell proliferation significantly but had a well-differentiated D1 cell phenotype that enhanced ALP expression in the early stage of cell cultures, specifically, at day 7. Results suggest that the SLA surface with anchored TTCP can accelerate progenitor bone cell mineralization. This study shows the potential clinical application of the constructed geometry in TTCP anchorage on Ti for dental implant surface modification.

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A new bioresorbable polylactide/calcium phosphate composite with improved mechanical strengths and a more basic filler, tetracalcium phosphate (TTCP), was prepared by melt compounding. N-(2-aminoethyl)-3-aminoproplytrimethoxysilane (AEAPS) and pyromellitic dianhydride (PMDA) were used to improve the interfacial adhesion between TTCP and polylactide (PLA). While AEAPS improved the dispersion of TTCP in the matrix, PMDA might react with the terminal hydroxyl group of PLA and the amino group on the surface of AEAPS modified TTCP, which could further enhance the interfacial strength. The tensile strength was improved to 68.4 MPa for the PLA/TTCP-AEAPS composite from 51.5 MPa for the PLA/TTCP composite (20 wt% of TTCP). Dynamic mechanical analysis suggested that there was a 51 % improvement in storage modulus compared to that of PLA alone, when PMDA (0.2 wt% of PMDA) was incorporated into the PLA/TTCP-AEAPS composite (5 wt% of TTCP). Using this new bioresorbable PLA composite incorporated with a more basic filler for biomedical application, the inflammation and allergic effect resulted from the degraded acidic product are expected to be reduced.

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In this study, calcium phosphate cements (CPC) were prepared by mixing cement powders of tetracalcium phosphate (TTCP) with a cement liquid of phosphate acid saline solution. Tetracycline (TTC)-CPC, chitosan-CPC and chitosan-TTC-CPC were investigated with different premixed schedule. It was demonstrate that both TTC and chitosan worked on the phase transition and crystal characteristics. TTCP mixed with phosphate acid saline solution had similar features of Fourier transform-infrared spectrometry (FT-IR) no matter it was mixed with chitosan or TTC or both. TTC premixed with cement liquid or powder had significant different features of FT-IR and 876 cm-1seemed to be a special peak for TTC when TTC was premixed with cement liquid. This was also supported by XRD analysis, which showed that TTC premixed with cement liquid improved phase transition of TTCP to OCP. Chitosan, as organic additive, regulates the regular crystal formation and inhibits the phase transition of TTCP to OCP, except when it is mingled with cement liquid premixed with TTC in field scanning electron microscope. It was concluded that the premixed schedule influences the crystal formation and phase transition, which may be associated with its biocompatibility and bioactivities in vivo.

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Research on calcium phosphate chemistry at NIST led to the discovery of the worlds first self-hardening calcium phosphate cements (CPC) in 1987. Laboratory, animal, and clinical studies were conducted to develop CPC into clinically useful biomaterials. The combination of self-hardening capability and high biocompatibility makes CPC a unique material for repairing bone defects. Near perfect adaptation of the cement to the tissue surfaces in a defect, and a gradual resorption followed by new bone formation are some of the other distinctive advantages of this biomaterial. In 1996 a CPC, consisting of tetracalcium phosphate and dicalcium phosphate anhydrous, was approved by the Food and Drug Administration (FDA) for repairing cranial defects in humans, thus becoming the first material of its kind available for clinical use. This paper will review the course of the development, the physical and chemical properties, and clinical applications of CPC.