The impact of the physicochemical properties of calcium phosphate ceramics on biocompatibility and osteogenic differentiation of mesenchymal stem cells.

OBJECTIVE: In this study we have focused on biocompatibility and osteoinductive capacity analysis of self-manufactured single-phase (HAP) and two-phase (HAP and ß-ТСР) bioactive ceramics with various chemical modifications (Fig. 1). RESULTS: We demonstrate a reduction in solubility for all analyzed composite after the treatment with H2O and H2O2, accompanied by an enhancement in adsorption activity. This modification also resulted in an increase in micro- and macroporosity, along with a rise in the open porosity. Adipose-derived mesenchymal stromal cells demonstrated excellent cell adhesion and survival when cultured with these ceramics. Calcium phosphate ceramics (H-500, HT-500, and HT-1 series) stimulated alkaline phosphatase expression, promoted calcium deposition, and enhanced osteopontin expression in ADSCs, independently inducing osteogenesis without additional osteogenic stimuli. These findings underscore the promising potential of HAP-based bioceramics for bone regeneration/reconstruction.

Efficacy of Self-assembling Peptide P11-4 in Remineralizing In Vitro Caries-like Lesions in Primary Enamel Samples in Combination with Calcium Phosphate-based Remineralization Agents.

Aims and background: The efficacy of self-assembling peptide P11-4 in combination with calcium-phosphate-based remineralization agents in remineralizing caries-like lesions in primary enamel was evaluated using a 21-day pH cycling experiment by Vickers microhardness [Vickers hardness number (VHN)] and scanning electron microscopy (SEM). Materials and methods: A total of 120 primary enamel samples were made to undergo a demineralization cycle to produce caries-like lesions. They were divided into six groups, namely negative control (NC), positive control (P11-4), and four interventional groups in which each of the following calcium-phosphate-based agents were used in combination with P11-4-calcium sucrose phosphate (CSP), bioactive glass (BG), casein phosphopeptides, and casein phosphopeptides with fluoride. A 21-day pH cycling experiment was carried out with alternating demineralization and remineralization phases. The enamel samples were analyzed at baseline, post production of caries-like lesions, and post 21-day pH cycling using Vickers microhardness and SEM. Results were statistically analyzed using repeated measures of analysis of variance (ANOVA), keeping the level of significance at 0.05. Results: Supplementing P11-4 with calcium-phosphate-based agents improved the surface hardness of the demineralized primary enamel samples, among which the fluoridated milk protein-based remineralization agent yielded a statistically significant improvement. Conclusion: P11-4 promoted the regeneration of incipient caries-like lesions. However, there is added benefit when this peptide is used in combination with a fluoridated calcium-phosphate-based agent. Clinical significance: This study would help the clinician compose an effective regimen for the patient to follow at home posttreatment with P11-4, in-office treatment. How to cite this article: Krishnamoorthi A, Shanbhog RS, Godhi BS, et al. Efficacy of Self-assembling Peptide P11-4 in Remineralizing In Vitro Caries-like Lesions in Primary Enamel Samples in Combination with Calcium Phosphate-based Remineralization Agents. Int J Clin Pediatr Dent 2024;17(5):552-557.

Biocompatibility and osteogenic assessment of experimental fluoride-doped calcium-phosphate cements on human dental pulp stem cells.

OBJECTIVES: This study investigated the impact of some specific experimental calcium phosphate cements doped with different fluoride salts (FDCPCs) concentrations on the basal functions of human Dental Pulp Stem Cells (hDPSCs). Furthermore, this study also examined the migration, as well as the mineralisation through osteogenic differentiation. METHODS: Experimental FDCPCs were formulated using different concentrations of calcium/sodium fluoride salts [(5 wt%: VS5F), (10 wt%: VS10F), (20 wt%: VS20F)]. A fluoride-free calcium phosphate (VS0F) was used as a control. The hDPSCs were assessed to evaluate their self-renewal and migration activity in the presence of eluates of the different FDCPCs. A viability assay in osteogenic conditions was carried out, along with the differentiation potential through Alkaline Phosphatase Activity (ALP), and Alizarin Red Staining (ARS). Moreover, the gene expression of specific markers (RUNX2, ALP, COL1α1, OCN, OPN, DSPP, MEPE, and DMP-1) was also evaluated. RESULTS: All the tested FDCPD had no influence on cell migrations, but they caused a decrease in cell viability in osteogenic conditions when not diluted. Conversely, the eluants of VS20F showed a positive effect on stem cell differentiation. This result was corroborated through ALP activity, ARS assay. Moreover, upregulation of specific gene markers such as RUNX2, DMP-1, and DSPP was observed in hDPSCs, especially when treated with VS20F. SIGNIFICANCE: The experimental FDCPC tested in this study exhibits a dose-dependent capacity to promote mineralisation in osteogenic environment. The FDCPC-VS20F seems to be the most promising experimental material suitable for developing of pulp-capping materials with osteogenic and bioactive properties.

Optimizing silicon doping levels for enhanced osteogenic and angiogenic properties of 3D-printed biphasic calcium phosphate scaffolds: An in vitro screening and in vivo validation study.

Biphasic calcium phosphate (BCP) ceramics are valued for their osteoconductive properties but have limited osteogenic and angiogenic activities, which restricts their clinical utility in bone defect repair. Silicon doping has emerged as an effective strategy to enhance these biological functions of BCP. However, the biological impact of BCP is influenced by the level of silicon doping, necessitating determination of the optimal concentration to maximize efficacy in bone repair. This study investigated the effects of silicon doping on both the physicochemical and biological properties of BCP, with a specific focus on osteogenic and angiogenic potentials. Results indicated that silicon doping exceeding 4 mol.% led to the formation of α-TCP, accelerating BCP degradation, enhancing silicon ion release, and promoting mineralization product formation. Simultaneously, silicon doping increased the porosity of BCP scaffolds, which typically reduces their compressive strength. Nevertheless, scaffolds doped with ≤4 mol.% silicon maintained compressive strengths exceeding 2 MPa. In vitro biological experiments indicated that higher levels of silicon doping (≥6 mol.%) partially inhibited the successful differentiation of stem cells and the vascularization of endothelial cells. Optimal conditions for promoting osteogenic differentiation and angiogenesis were identified between 2 and 4 mol.% silicon doping, with an optimal level of approximately 4 mol.%. Subsequent in vivo experiments confirmed that BCP scaffolds doped with 4 mol.% silicon effectively promoted vascularization and new bone formation, highlighting their potential for clinical bone defect repair.

Biodegradable WE43 Mg alloy/hydroxyapatite interpenetrating phase composites with reduced hydrogen evolution.

Biodegradable magnesium implants offer a solution for bone repair without the need for implant removal. However, concerns persist regarding peri-implant gas accumulation, which has limited their widespread clinical acceptance. Consequently, there is a need to minimise the mass of magnesium to reduce the total volume of gas generated around the implants. Incorporating porosity is a direct approach to reducing the mass of the implants, but it also decreases the strength and degradation resistance. This study demonstrates that the infiltration of a calcium phosphate cement into an additively manufactured WE43 Mg alloy scaffold with 75 % porosity, followed by hydrothermal treatment, yields biodegradable magnesium/hydroxyapatite interpenetrating phase composites that generate an order of magnitude less hydrogen gas during degradation than WE43 scaffolds. The enhanced degradation resistance results from magnesium passivation, allowing osteoblast proliferation in indirect contact with composites. Additionally, the composites exhibit a compressive strength 1.8 times greater than that of the scaffolds, falling within the upper range of the compressive strength of cancellous bone. These results emphasise the potential of the new biodegradable interpenetrating phase composites for the fabrication of temporary osteosynthesis devices. Optimizing cement hardening and magnesium passivation during hydrothermal processing is crucial for achieving both high compressive strength and low degradation rate.

Injectable calcium phosphate cement integrated with BMSCs-encapsulated microcapsules for bone tissue regeneration.

Injectable calcium phosphate cement (CPC) offers significant benefits for the minimally invasive repair of irregular bone defects. However, the main limitations of CPC, including its deficiency in osteogenic properties and insufficient large porosity, require further investigation and resolution. In this study, alginate-chitosan-alginate (ACA) microcapsules were used to encapsulate and deliver rat bone mesenchymal stem cells (rBMSCs) into CPC paste, while a porous CPC scaffold was established to support cell growth. Our results demonstrated that the ACA cell microcapsules effectively protect the cells and facilitate their transport into the CPC paste, thereby enhancing cell viability post-implantation. Additionally, the ACA+CPC extracts were found to stimulate osteogenic differentiation of rBMSCs. Furthermore, results from a rat cranial parietal bone defect model showed that ACA microcapsules containing exogenous rBMSCs initially improved the in situ osteogenic potential of CPC within bone defects, providing multiple sites for bone growth. Over time, the osteogenic potential of the exogenous cells diminishes, yet the pores created by the microcapsules persist in supporting ongoing bone formation by recruiting endogenous cells to the osteogenic sites. In conclusion, the utilization of ACA loaded stem cell microcapsules satisfactorily facilitate osteogenesis and degradation of CPC, making it a promising scaffold for bone defect transplantation.

Delivery of extracellular vesicles loaded with immune checkpoint inhibitors for immunotherapeutic management of glioma.

Glioma is a common primary malignant brain tumor with low survival rate. Immunotherapy with immune checkpoints inhibitors (ICI) can be a choice for glioma management, and extracellular vesicles (EVs) are recognized as a potential drug delivery system for various disease management due to their enhanced barrier permeation ability and immunomodulatory effect. The aim of this study is to develop ICI-loaded EVs (ICI/EV) that have sufficient efficacy in managing glioma. Calcium phosphate particles (CaP) were used to stimulate the secretion of EVs from murine macrophage cells. CaP conditioning of cells showed an enhanced amount of EVs secretion and macrophage polarization toward a proinflammatory phenotype. The CaP-induced EVs were shown to polarize macrophages into proinflammatory phenotype in vitro, as correlated with the conditioning method. ICI/EVs were successfully prepared with high loading efficiency using the sonication method. The EVs can be distributed throughout the entire brain upon intranasal administration and facilitate ICIs distribution into glioma lesion. Combinatory treatment with ICI/EVs showed benefit in glioma-bearing mice by reducing their tumor volume and prolonging their survival. Cytotoxic T cell infiltration, polarization of tumor-associated macrophage, and lower tumor proliferation were observed in ICI/EVs-treated mice. The developed ICI/EVs showed promise in immunotherapeutic management of glioma.

Biomimetic synthetic test system based on hydroxyapatite cement for adhesive strength evaluation of experimental mineral-organic bone adhesive materials.

The development of bone adhesive materials is a research field of high relevance for the advancement of clinical procedures. Despite this, there are currently no material candidates meeting the full range of requirements placed on such a material, such as biocompatibility, sufficient mechanical properties and bond strength under biological conditions, practical applicability in a clinical setting, and no adverse effect on the healing process itself. A serious obstacle to the advancement of the field is a lack in standardized methodology leading to comparable results between experiments and different research groups. Natural bone samples are the current gold-standard material used to perform adhesive strength experiments, however they come with a number of drawbacks, including high sample variability due to unavoidable natural causes and the impossibility to reliably recreate test conditions to repeat experiments. This paper introduces a valuable auxiliary test method capable of producing large numbers of synthetic test specimens which are chemically similar to bone and can be produced in different laboratories so to repeat experiments under constant conditions across laboratories. The substrate is based on a hydroxyapatite forming cement with addition of gelatine as organic component. Crosslinking of the organic component is performed to improve mechanical properties. In order to demonstrate the performance of the developed method, various experimental and commercial bone/tissue adhesive materials were tested and compared with results obtained by established methods to highlight the potential of the test system.

Biomimetic Scaffolds of Calcium-Based Materials for Bone Regeneration.

Calcium-based materials, such as calcium carbonate, calcium phosphate, and calcium silicate, have attracted significant attention in biomedical research, owing to their unique physicochemical properties and versatile applications. The distinctive characteristics of these materials, including their inherent biocompatibility and tunable structures, hold significant promise for applications in bone regeneration and tissue engineering. This review explores the biomedical applications of calcium-containing materials, particularly for bone regeneration. Their remarkable biocompatibility, tunable nanostructures, and multifaceted functionalities make them pivotal for advancing regenerative medicine, drug delivery system, and biomimetic scaffold applications. The evolving landscape of biomedical research continues to uncover new possibilities, positioning calcium-based materials as key contributors to the next generation of innovative biomaterial scaffolds.

Calcium phosphate complex of recombinant human thrombomodulin promote bone formation in interbody fusion.

Interbody fusion is an orthopedic surgical procedure to connect two adjacent vertebrae in patients suffering from spinal disc disease. The combination of synthetic bone grafts with protein-based drugs is an intriguing approach to stimulate interbody bone growth, specifically in patients exhibiting restricted bone progression. Recombinant human thrombomodulin (rhTM), a novel protein drug characterized by its superior stability and potency, shows promise in enhancing bone formation. A composite bone graft, termed CaP-rhTM, has been synthesized, combining calcium phosphate (CaP) microparticles as a delivery vehicle for rhTM to facilitate interbody fusion. In vitro studies have demonstrated that rhTM significantly promotes the proliferation and maturation of preosteoblasts at nanogram dosage, while exerting minimal impact on osteosarcoma cell growth. The expression levels of mature osteoblast markers, including osteocalcin, osteopontin, alkaline phosphatase, and calcium deposition were also enhanced by rhTM. In rat caudal disc model of interbody fusion, CaP-rhTM with 800 ng of drug doaeage was implanted along with a polylactic acid (PLA) cage, to ensure structural stability within the intervertebral space. Microcomputed tomography analyses revealed that from 8 to 24 weeks, CaP-rhTM substantially improves both bone volume and trabecular architecture, in addition to the textural integrity of bony endplate surfaces. Histological examination confirmed the formation of a continuous bone bridge connecting adjacent vertebrae. Furthermore, biomechanical assessment via three-point bending tests indicated an improved bone quality of the fused disc. This study has demostrated that rhTM exhibits considerable potential in promoting osteogenesis. The use of CaP-rhTM has also shown significant improvements in promoting interbody fusion.

In-vitro and in-vivo comparative studies of treatment effects on enamel demineralization during orthodontic therapy: implications for clinical early-intervention strategy.

OBJECTIVES: This study aimed to investigate if CPP-ACP / infiltrating resin was superior in treating enamel demineralization during orthodontic therapy compared with fluoride varnish, in order to provide early-intervention implications for dental professionals. MATERIALS AND METHODS: In the in-vitro study, premolars were grouped into four: remineralization with fluoride varnish / CPP-ACP, sealing with infiltrating resin, and negative control. Experimental demineralization of enamel surfaces was analyzed using techniques of QLF, SEM, EDS and micro-hardness testing. An in-vivo intervention study was conducted on patients randomly assigned into three groups. At the baseline and every-3-month follow-up, QLF parameters were compared temporally and parallelly to yield potential implications for promotion in clinical practice. RESULTS: The in-vitro study performed on 48 experimental tooth surfaces demonstrated that sealing with infiltrating resin reduced enamel surface porosity and increased surface micro-hardness significantly. In the in-vivo intervention study on 163 tooth surfaces, it was suggested that for those who meet the criteria of -10 < ΔF < -6 and - 1000 < ΔQ < -20 at the baseline, all these treatment methods could achieve acceptable outcomes; with the rising of absolute values of ΔF and ΔQ, sealing with infiltrating resin showed more evident advantages. CONCLUSION: For enamel demineralization during orthodontic therapy, all the treatment methods involved in this study showed acceptable effectiveness but had respective characteristics in treatment effects. QLF parameters could be used as indicators for clinical early-intervention strategy with regards to this clinical issue. CLINICAL RELEVANCE: With QLF parameters, clinical early-intervention strategy for enamel demineralization during orthodontic therapy could be optimized.

Fabrication of In Situ-Cross-Linked N-Succinyl Chitosan/Oxidized Alginate Hydrogel-Loaded Ascorbic Acid and Biphasic Calcium Phosphate for Bone Tissue Engineering.

Bone tissue engineering is a promising technology being studied globally to become an effective and sustainable method to treat the problems of damaged or diseased bones. In this work, we developed an in situ cross-linking hydrogel system that combined N-succinyl chitosan (NSC) and oxidized alginate (OA) at varying mixing ratios through Schiff base cross-linking. The hydrogel system also contains biphasic calcium phosphate (BCP) and ascorbic acid (AA), which could enhance biological characteristics and accelerate bone repair. The hydrogels' properties were examined through physicochemical tests such as scanning electron microscopy (SEM), energy-dispersive x-ray spectroscopy (EDS), Fourier transform infrared spectroscopy (FT-IR), x-ray diffraction (XRD), pore size and porosity measurement, swelling ratio, degradation rate, AA release study, as well as cytocompatibility, including live/dead and cytotoxicity assays. The results revealed that the supplementation of AA and BCP components can affect the physico-mechanical properties of the hydrogel system. However, they exhibited noncytotoxic properties. Overall, the results demonstrated that the hydrogel composed of 3% (w/v) NSC and 3% (w/v) OA (NSC: OA volume ratio is 8:2) loaded with 40% (w/w) BCP and 0.3 mg/mL AA has the potential for bone regeneration.

Greater urinary calcium excretion is associated with diminished bone accrual in youth with type 1 diabetes.

CONTEXT: The adverse skeletal effects of type 1 diabetes (T1D) include deficient bone accrual and lifelong increased fracture risk. The contributors to impaired bone accrual in people with T1D are incompletely understood. OBJECTIVE: To determine if urinary calcium excretion is associated with impaired bone accrual in youth with T1D and to characterize the contribution of glycemic control and markers of bone mineral metabolism to urinary calcium excretion. DESIGN: Observational study. PARTICIPANTS: 50 participants with T1D aged 6-20 years completed a 12-month longitudinal study of bone accrual. A second cohort of 99 similarly aged participants with T1D completed cross-sectional 24-hr urine and blood collections. MAIN OUTCOME MEASURE: Whole body less head bone mineral content (WBLH BMC) velocity Z-score and fractional excretion of calcium (FeCa). RESULTS: Participants in the bone accrual cohort had lower WBLH BMC velocity compared to a healthy reference dataset (Z-score -0.3 ± 1.0, p=0.03). FeCa was negatively associated with WBLH BMC velocity Z-score, ρ= -0.47, p=0.001. In the urinary calcium excretion cohort, intact parathyroid hormone (ß= -0.4, p=0.01), beta c-telopeptide (ß=0.35, p=0.007), and either hemoglobin A1c (HbA1c, ß=0.08, p=0.03) or urine fractional glucose excretion (ß=0.07, p=0.03) were associated with FeCa in multivariable regression models that included known determinants of urinary calcium excretion. CONCLUSIONS: Urinary calcium excretion was negatively associated with bone accrual in this cohort of youth with T1D. Mechanistic studies are needed to determine if interventions to reduce urinary calcium excretion could increase bone accrual and reduce skeletal fragility in people with T1D.

Hybrid Hydroxyapatite-Metal Complex Materials Derived from Amino Acids and Nucleobases.

Calcium phosphates (CaPs) and their substituted derivatives encompass a large number of compounds with a vast presence in nature that have aroused a great interest for decades. In particular, hydroxyapatite (HAp, Ca10(OH)2(PO4)6) is the most abundant CaP mineral and is significant in the biological world, at least in part due to being a major compound in bones and teeth. HAp exhibits excellent properties, such as safety, stability, hardness, biocompatibility, and osteoconductivity, among others. Even some of its drawbacks, such as its fragility, can be redirected thanks to another essential feature: its great versatility. This is based on the compound's tendency to undergo substitutions of its constituent ions and to incorporate or anchor new molecules on its surface and pores. Thus, its affinity for biomolecules makes it an optimal compound for multiple applications, mainly, but not only, in biological and biomedical fields. The present review provides a chemical and structural context to explain the affinity of HAp for biomolecules such as proteins and nucleic acids to generate hybrid materials. A size-dependent criterium of increasing complexity is applied, ranging from amino acids/nucleobases to the corresponding macromolecules. The incorporation of metal ions or metal complexes into these functionalized compounds is also discussed.

Do Stainless-Steel Pins Coated with Fibroblast Growth Factor-Calcium Phosphatase Composite Layers Have Anti-Infective Effects?

Background: The most problematic complication of external fixation is infection at the pin insertion site. Technology that improves the adhesion of the external fixation pin to the skin, subcutaneous tissue, and bone may prevent infection at the pin site. The purpose of this study is to formulate a calcium phosphate-fibroblast growth factor (Cp-FGF) coating on a stainless-steel external fixation pin and to verify its effectiveness in reducing infection at the pin site and its possible influence on bone fixation in animal experiments. Methods: We compared stainless-steel screws without coating (SS group; n = 32), those with a calcium phosphate coating (Cp group; n = 30), those with a Cp-FGF coating (FGF group; n = 32), and those with a Cp-FGF coating having enhanced biological activity (FGF+ group; n = 32) in male Japanese white domesticated rabbits. Screws were inserted percutaneously into the bilateral proximal tibial diaphysis of the rabbits and implanted for 4 weeks. Screws and periscrew tissue were observed postoperatively for qualitatively assessing infection. Results: Infection assessment by gross findings after 4 weeks (at screw removal) showed no significant differences between the groups. Histopathological evaluation of soft tissue infection and bone tissue infection showed no significant differences between the groups for either soft tissue or bone tissue. Since neither the FGF+ group nor the FGF group showed anti-infective effects, the biological activity of FGF is not the only determining factor. We compared SEM, XRD, coating detaching test, sustained release test, and bioassay to examine physicochemical properties among the coatings but found no sufficient differences. Conclusions: It is suggested that improving the tissue adhesion to and/or biocompatibility of pins is also important to improve the in vivo performance of Cp-FGF-coated external fixation pins.

Local mRNA Delivery from Nanocomposites Made of Gelatin and Hydroxyapatite Nanoparticles.

Local delivery of messenger ribonucleic acid (mRNA) is increasingly being advocated as a promising new strategy to enhance the performance of biomaterials. While extensive research has been dedicated to the complexation of these oligonucleotides into nanoparticles to facilitate systemic delivery, research on developing suitable biomaterial carriers for the local delivery of mRNA is still scarce. So far, mRNA-nanoparticles (mRNA-NPs) are mainly loaded into traditional polymeric hydrogels. Here, we show that calcium phosphate nanoparticles can be used for both reinforcement of nanoparticle-based hydrogels and the complexation of mRNA. mRNA was incorporated into lipid-coated calcium phosphate nanoparticles (LCPs) formulated with a fusogenic ionizable lipid in the outer layer of the lipid coat. Nanocomposites of gelatin and hydroxyapatite nanoparticles were prepared at various ratios. Higher hydroxyapatite nanoparticle content increased the viscoelastic properties of the nanocomposite but did not affect its self-healing ability. Combination of these nanocomposites with peptide, lipid, and the LCP mRNA formulations achieved local mRNA release as demonstrated by protein expression in cells in contact with the biomaterials. The LCP-based formulation was superior to the other formulations by showing less sensitivity to hydroxyapatite and the highest cytocompatibility.

Multiparametric influence of 3D-printed organo-mineral scaffolds on bone regeneration.

The development of synthetic bone substitutes that equal or exceed the efficacy of autologous graft remains challenging. In this study, a rat calvarial defect model was used as a reference to investigate the influence of composition and architecture of 3D-printed cement, with or without bioactives, on tissue regeneration. Printable cement pastes were formulated by combining hyaluronic acid and cement precursors. Cementitious scaffolds were printed with 3 different patterns. After 7 weeks of implantation with or without bone marrow, multiparametric qualitative and quantitative assessments were performed using µCT, SEM, and histology. None of the set-up strategies was as efficient as autologous cancellous bone graft to repair calvarial defects. Nonetheless, the presence of scaffold improved the skull vault closure, particularly when the scaffold was soaked in total bone marrow before implantation. No significant effect of scaffold macro-architecture was observed on tissue mineralization. Magnesium phosphate-based scaffolds (MgP) seemed to induce higher bone formation than their calcium-phosphate-based counterparts. They also displayed a quicker biodegradation and sparse remaining material was found after 7 weeks of implantation. Although further improvements are required to reach clinical settings, this study demonstrated the potential of organo-mineral cements for bone regeneration and highlighted the peculiar properties of MgP-based cements.

Promotion effect of proanthocyanidin on dentin remineralization via the polymer induced liquid precursor process.

Proanthocyanidin (PA) has demonstrated promise as a dental biomodifier for maintaining dentin collagen integrity, yet there is limited evidence regarding its efficacy in dentin repair. The aim of this study was to investigate the effect of PA on dentin remineralization through the polymer induced liquid precursor (PILP) process, as well as to assess the mechanical properties of the restored dentin. Demineralized dentin was treated with a PA-contained remineralization medium, resulting in the formation of PA-amorphous calcium phosphate (ACP) nanoparticles via the PILP process. The kinetics and microstructure of remineralized dentin were examined through the use of Fourier transform infrared spectroscopy(FTIR), attenuated total reflectance-FTIR, scanning electron microscopy, transmission electron microscopy. The results showed that the application of PA facilitated the process of dentin remineralization, achieving completion within 48 h, demonstrating a notable reduction in time required. Following remineralization, the mechanical properties of the dentin exhibited an elastic modulus of 15.89 ± 1.70 GPa and a hardness of 0.47 ± 0.08 GPa, which were similar to those of natural dentin. These findings suggest that combining PA with the PILP process can promote dentin remineralization and improve its mechanical properties, offering a promising new approach for dentin repair in clinical practice.

Incorporation of calcium phosphate cement into decellularized extracellular matrix enhances its bone regenerative properties.

Decellularized extracellular matrix (dECM) hydrogels are engineered constructs that are widely-used in the field of regenerative medicine. However, the development of ECM-based hydrogels for bone tissue engineering requires enhancement in its osteogenic properties. For this purpose, we initially employed bone-derived dECM hydrogel (dECM-Hy) in combination with calcium phosphate cement (CPC) paste to improve the biological and structural properties of the dECM hydrogel. A decellularization protocol for bovine bone was developed to prepare dECM-Hy, and the mechanically-tuned dECM/CPC-Hy was built based on both rheological and mechanical characteristics. The dECM/CPC-Hy displayed a double swelling ratio and compressive strength. An interconnected structure with distinct hydroxyapatite crystals was evident in dECM/CPC-Hy. The expression levels of Alp, Runx2 and Ocn genes were upregulated in dECM/CPC-Hy compared to the dECM-Hy. A 14-day follow-up of the rats receiving subcutaneous implanted dECM-Hy, dECM/CPC-Hy and mesenchymal stem cells (MSCs)-embedded (dECM/CPC/MSCs-Hy) showed no toxicity, inflammatory factor expression or pathological changes. Radiography and computed tomography (CT) of the calvarial defects revealed new bone formation and elevated number of osteoblasts-osteocytes and osteons in dECM/CPC-Hy and dECM/CPC/MSCs-Hy compared to the control groups. These findings indicate that the dECM/CPC-Hy has substantial potential for bone tissue engineering.

Mn2+ vs Co2+ substitution into ß-TCP: Structural details and bone cells response.

This work investigated the range of substitution of two biologically relevant ions, namely Mn2+ and Co2+, into the structure of ß-tricalcium phosphate, as well as their influence on bone cells response. To this aim, ß-TCP was synthesized by solid state reaction in the presence of increasing amount of the substituent ions. The results of the X-ray diffraction analysis reveal that just limited amounts of these ions can enter into the ß-TCP structure: 15 at% and 20 at% for cobalt and manganese, respectively. Substitution provokes aggregation of the micrometric particles and reduction of the lattice constants. In particular, the dimension of the c-parameter exhibits a discontinuity at about 10 at% for both cations, although with different trend. Moreover, Rietveld refinement demonstrates a clear preference of both manganese and cobalt for the octahedral site (V). The influence of these ions on cell response was tested on osteoblast, osteoclast and endothelial cells. The results indicate that the presence of manganese promotes a good osteoblast viability, significantly enhances the expression of osteoblast key genes and the angiogenic process of endothelial cells, while inhibiting osteoclast resorption. At variance, osteoblast viability appears reduced in the presence of Co samples, on which osteoblast genes reach higher expression than on ß-TCP just in a few cases. On the other hand, the results clearly show that cobalt significantly stimulates the angiogenic process and inhibits osteoclast resorption.