Calcium Phosphate Cement Scaffolds Research Articles

Self-setting calcium phosphate cement scaffolds with pre-forming and in-situ forming interconnected macropores: Comparative study in vitro and in vivo

Creating interconnected macropores in calcium phosphate cement (CPC) is an effective strategy to promote its degradation and osteogenesis. However, little attention has been given to the osteogenic effect of the CPC scaffolds with pre-forming and in-situ forming interconnected macropores. Herein, two types of CPC scaffolds were prepared by infiltrating CPC pastes into 3D-printed polycaprolactone (PCL) and poly(lactic-co-glycolic acid) (PLGA) networks. Meanwhile, the sacrificial PCL network within CPC was dissolved to obtain the CPC scaffold with approximately 300 μm macropores, whereas the PLGA network was retained within the CPC to obtain the PLGA/CPC scaffold. The results indicated that the PLGA/CPC scaffold showed higher degradation rate and compressive strength compared to the CPC scaffold with pre-forming interconnected macropores. However, the proliferation rate and alkaline phosphatase activity of mouse bone mesenchymal stem cells cultured with the CPC scaffold were superior to those cultured with the PLGA/CPC scaffold. In addition, greater formation of new bone and material degradation in the CPC scaffold compared to the PLGA/CPC scaffold after implanted in the rabbit femoral defects.

A cross-sectional study of the anatomy of the jaws of a central-European caucasian population using cone beam computer tomography as a prerequisite for designing pre-formed calcium phosphate cement scaffolds

PurposeThis study aims to measure the cortical and cancellous bone thickness in the upper and lower jaws, serving as a data template for developing pre-defined calcium phosphate cement primary implant forms. These measurements are crucial for creating a biphasic scaffold. MethodsForty complete jaws were assessed for cortical bone shape and thickness using statistical analysis and specific software tools. Sex and age were considered, and four groups were created. ResultsThe cumulative thickness of the cortical layer varied from region to region. In both the upper and lower jaws, the cortical layer in the molar region was significantly thicker than in the frontal region. Within the alveolar process, cortical thickness increases with distance from the alveolar crest on both sides. The oral side of the lower jaw is significantly thicker than the vestibular side. For the upper jaw, no significant differences between the oral and vestibular sides were found in this study. Additionally, it is noteworthy that men have a significantly thicker cortical layer than women. Regarding age, no significant overall differences were found. ConclusionMathematical analysis of anatomical forms using polynomial functions improves understanding of jaw anatomy. This approach facilitates the design of patient-specific scaffold structures, minimizing the need for costly and time-consuming planning and enabling more efficient implementation of optimal therapy.

Synergistic effects of calcium silicate/zinc silicate dual compounds and in-situ interconnected pores on promoting bone regeneration of composite scaffolds

Rapid bone regeneration in implants is important for successful transplantation. In this regard, we report the development of calcium silicate/zinc silicate (CS/ZS) dual-compound-incorporated calcium phosphate cement (CPC) scaffolds with a three-dimensional poly (lactic-co-glycolic acid) network that synergistically promote bone regeneration. In vitro results demonstrated that the incorporation of CS/ZS dual compounds into the CPC significantly promoted the osteogenic differentiation of stem cells compared to the addition of CS or ZS alone. Moreover, the bone-regeneration efficacy of the composite scaffolds was validated by filling in femur condyle defects in rabbits, which showed that the scaffolds with CS and ZS possessed a great bone repair effect, as evidenced by more new bone formation and a faster scaffold biodegradation compared to the scaffold with CS alone.

Injectable periodontal ligament stem cell-metformin-calcium phosphate scaffold for bone regeneration and vascularization in rats.

Injectable and self-setting calcium phosphate cement scaffold (CPC) capable of encapsulating and delivering stem cells and bioactive agents would be highly beneficial for dental and craniofacial repairs. The objectives of this study were to: (1) develop a novel injectable CPC scaffold encapsulating human periodontal ligament stem cells (hPDLSCs) and metformin (Met) for bone engineering; (2) test bone regeneration efficacy in vitro and in vivo. hPDLSCs were encapsulated in degradable alginate fibers, which were then mixed into CPC paste. Five groups were tested: (1) CPC control; (2) CPC +hPDLSC-fibers +0% Met (CPC + hPDLSCs + 0%Met); (3) CPC +hPDLSC-fibers +0.1% Met (CPC + hPDLSCs + 0.1%Met); (4) CPC +hPDLSC-fibers +0.2% Met (CPC + hPDLSCs + 0.2%Met); (5) CPC +hPDLSC-fibers +0.4% Met (CPC + hPDLSCs + 0.4%Met). The injectability, mechanical properties, metformin release, and hPDLSC osteogenic differentiation and bone mineral were determined in vitro. A rat cranial defect model was used to evaluate new bone formation. The novel construct had good injectability and physical properties. Alginate fibers degraded in 7 days and released hPDLSCs, with 5-fold increase of proliferation (p＜0.05). The ALP activity and mineral synthesis of hPDLSCs were increased by Met delivery (p＜0.05). Among all groups, CPC+hPDLSCs+0.1%Met showed the greatest cell mineralization and osteogenesis, which were 1.5-10folds those without Met (p＜0.05). Compared to CPC control, CPC+hPDLSCs+0.1%Met enhanced bone regeneration in rats by 9 folds, and increased vascularization by 3 folds (p＜0.05). The novel injectable construct with hPDLSC and Met encapsulation demonstrated excellent efficacy for bone regeneration and vascularization in vivo in an animal model. CPC+hPDLSCs+0.1%Met is highly promising for dental and craniofacial applications.

Development of 3D Printable Calcium Phosphate Cement Scaffolds with Cockle Shell Powders.

Three-dimensional (3D) printed calcium phosphate cement (CPC) scaffolds are increasingly being used for bone tissue repair. Traditional materials used for CPC scaffolds, such as bovine and porcine bone, generally contain low amounts of calcium phosphate compounds, resulting in reduced production rates of CPC scaffolds. On the other hand, cockle shells contain more than 99% CaCO3 in the form of amorphous aragonite with excellent biocompatibility, which is expected to increase the CPC production rate. In this study, 3D-printed cockle shell powder-based CPC (CSP-CPC) scaffolds were developed by the material extrusion method. Lactic acid and hyaluronic acid were used to promote the printability. The characterization of CSP-CPC scaffolds was performed using Fourier transform infrared spectra, X-ray diffraction patterns, and scanning electron microscopy. The biocompatibility of CSP-CPC scaffolds was evaluated using cell viability, Live/Dead, and alkaline phosphatase assays. In addition, CSP-CPC scaffolds were implanted into the mouse calvarial defect model to confirm bone regeneration. This study provides an opportunity to create high value added in fishing villages by recycling natural products from marine waste.

Calvaria defect regeneration via human periodontal ligament stem cells and prevascularized scaffolds in athymic rats

BackgroundVascularization plays an important role in dental and craniofacial regenerations. Human periodontal ligament stem cells (hPDLSCs) are a promising cell source and, when co-cultured with human umbilical vein endothelial cells (hUVECs), could promote vascularization. The objectives of this study were to develop a novel prevascularized hPDLSC-hUVEC-calcium phosphate construct, and investigate the osteogenic and angiogenic efficacy of this construct with human platelet lysate (hPL) in cranial defects in rats for the first time. MethodshPDLSCs and hUVECs were co-cultured on calcium phosphate cement (CPC) scaffolds with hPL. Cell proliferation, angiogenic gene expression, angiogenesis, alkaline phosphatase activity, and cell-synthesized minerals were determined. Bone and vascular regenerations were investigated in rat critical-sized cranial defects in vivo. ResultshPDLSC-hUVEC-CPC-hPL group had 2-fold greater angiogenic expressions and cell-synthesized mineral synthesis than hPDLSC-hUVEC-CPC group (p < 0.05). Microcapillary-like structures were formed on scaffolds in vitro. hPDLSC-hUVEC-CPC-hPL group had more vessels than hPDLSC-hUVEC-CPC group (p < 0.05). In cranial defects in rats, hPDLSC-hUVEC-CPC-hPL group regenerated new bone amount that was 2.1 folds and 4.0 folds, respectively, that of hPDLSC-hUVEC-CPC group and CPC control (p < 0.05). New blood vessel density of hPDLSC-hUVEC-CPC-hPL group was 2 folds and 7.9 folds, respectively, that of hPDLSC-hUVEC-CPC group and CPC control (p < 0.05). ConclusionThe hPL pre-culture method is promising to enhance bone regeneration via prevascularized CPC. Novel hPDLSC-hUVEC-CPC-hPL prevascularized construct increased new bone formation and blood vessel density by 4–8 folds over CPC control. Clinical significanceNovel hPDLSC-hUVEC-hPL-CPC prevascularized construct greatly increased bone and vascular regeneration in vivo and hence is promising for a wide range of craniofacial applications.

Investigation of manufacturing deviations of CPC scaffolds for improving the design process

Abstract In current implantology, calcium phosphate cement (CPC) is increasingly used. A special focus is given to CPC scaffolds, as they are suitable for cell settlement and growth due to their positive osteoconductive properties. The design of the scaffolds is of decisive importance for this. The value ranges of the geometric parameters of these scaffolds (e.g. path distance, path diameter), which are positive for both cell settlement and cell growth, are very small. Manufacturing deviations therefore have a significant impact on cell settlement and growth. The pasty manufacturing consistency can cause sagging at the path interstices of a layer immediately below, resulting in significant manufacturing deviations. A larger path distance and thus a larger path interstitial space promotes cell settlement, but at the same time increases the risk of CPC path shape inconsistency. This in turn can have a negative effect on cell settlement. Therefore, the aim of this paper is to investigate the discrepancies between the nominal and actual state at a path distance favourable to cell settlement on the basis of manufactured CPC scaffolds. In this context, geometric and manufacturing parameters of the shape deviation are to be identified and constructive design adaptations are to be derived on the basis of these. In addition, the effects of the shape and position deviations on the flow behaviour will be investigated.

About the Mechanical Strength of Calcium Phosphate Cement Scaffolds

For the treatment of bone defects, biodegradable, compressive biomaterials are needed as replacements that degrade as the bone regenerates. The problem with existing materials has either been their insufficient mechanical strength or the excessive differences in their elastic modulus, leading to stress shielding and eventual failure. In this study, the compressive strength of CPC ceramics (with a layer thickness of more than 12 layers) was compared with sintered β-TCP ceramics. It was assumed that as the number of layers increased, the mechanical strength of 3D-printed scaffolds would increase toward the value of sintered ceramics. In addition, the influence of the needle inner diameter on the mechanical strength was investigated. Circular scaffolds with 20, 25, 30, and 45 layers were 3D printed using a 3D bioplotter, solidified in a water-saturated atmosphere for 3 days, and then tested for compressive strength together with a β-TCP sintered ceramic using a Zwick universal testing machine. The 3D-printed scaffolds had a compressive strength of 41.56 ± 7.12 MPa, which was significantly higher than that of the sintered ceramic (24.16 ± 4.44 MPa). The 3D-printed scaffolds with round geometry reached or exceeded the upper limit of the compressive strength of cancellous bone toward substantia compacta. In addition, CPC scaffolds exhibited more bone-like compressibility than the comparable β-TCP sintered ceramic, demonstrating that the mechanical properties of CPC scaffolds are more similar to bone than sintered β-TCP ceramics.

Effect of inoculation density of bone marrow mesenchymal stem cells cultured on calcium phosphate cement scaffold on osteogenic differentiation.

Calcium phosphate cements (CPCs) are biocompatible materials that have been evaluated as scaffolds in bone tissue engineering. At present, the stem cell density of inoculation on CPC scaffold varies. The aim of this study is to analyze the effect of seeding densities on cell growth and osteogenic differentiation of bone marrow mesenchymal stem cells (BMMSCs) on a calcium phosphate cements (CPCs) scaffold. BMMSCs derived from minipigs were seeded onto a CPC scaffold at three densities [1million/mL (1M), 5 million/mL (5M) and 25million/mL 25M)], and cultured for osteogenic induction for 1, 4 and 8 days. Well adhered and extended BMMSCs on the CPC scaffold showed significantly different proliferation rates within each seeding density group at different time points (P <0.05). The number of live cells per unit area in 1M, 5M and 25M increased by 3.5, 3.9 and 2.5 folds respectively. The expression of ALP peaked at 4 days post inoculation with the fold-change being 2.6 and 2.8 times higher in 5M and 25M respectively as compared to 1M. The expression levels of OC, Coll-1 and Runx-2 peaked at 8 days post inoculation. An optimal seeding density may be more conducive for cell proliferation, differentiation, and extracellular matrix synthesis on scaffolds. We suggest the optimal seeding density should be 5million/mL.

Icariside II-Loaded Calcium Phosphate Cement Scaffolds Combined with Bone Marrow Mesenchymal Stem Cells for Osteogenesis

Purpose: Previous studies have demonstrated the osteogenic effects of icariside II (ICSII), which is a metabolic product of the prenylated active flavonol icariin (ICA) from the roots of Epimedium. However, the in vivo osteogenic effects of ICSII remain unclear. Thus, in this study, we evaluated the osteogenic effects of ICSII in vivo. Methods: Complexes of calcium phosphate cement (CPC) and bone marrow mesenchymal stem cells (BMSCs) with or without ICSII were subcutaneously implanted into nude mice (ectopic osteogenesis test) and into tooth sockets in beagles after maxillary canine tooth extraction (in situ osteogenesis test). The samples were harvested at different time points, and the in vivo osteogenic effect of the ICSII on the BMSCs was evaluated by histology, microcomputed tomography (micro-CT) and the bone mineralization apposition rate (MAR). Results: The proliferation and viability of BMSCs in the ICSII-loaded CPC scaffold were significantly increased (P &lt; 0.01). The new bone area and MAR in the CPC+BMSC+ICSII group were greater than those in the CPC+BMSC group (P &lt; 0.05), but there was no significant difference in markers evaluated by immunohistochemistry and integrated optical density (IOD) analysis (P &gt; 0.05), with the exception of Runx-2 expression in the CPC+BMSC+ICSII group. After 2 months, the bone mineral content (BMC) and specific bone surface (bone surface divided by bone volume, BS/BV) were significantly increased (P &lt; 0.05) in the CPC+BMSC+ICSII group compared with the CPC+BMSC group. The bone mineral density (BMD), bone volume divided by total volume (BV/TV), and trabecular number (Tb.N) were increased in the CPC+BMSC+ICSII group, but the differences were not significant (P &gt; 0.05). Conclusions: Our results suggest that ICSII can likely induce bone formation by BMSCs and be used as a promising factor for building scaffold composites in bone tissue engineering.

3D extrusion printing of density gradients by variation of sinusoidal printing paths for tissue engineering and beyond.

During extrusion printing of pasty biomaterials, internal geometries are mainly adjusted by positioning of straightly deposited strands which does not allow realization of spatially adaptable density gradients in x-, y- and z-direction for anisotropic scaffolds or anatomically shaped constructs. Herein, an alternative concept for printing patterns based on sinusoidal curves was evaluated using a clinically approved calcium phosphate cement (CPC). Infill density in scaffolds was adjusted by varying wavelength and amplitude of a sinus curve. Both wavelength and amplitude factors were defined by multitudes of the applied nozzle diameter. For CPC as a biomaterial ink in bone application, porosity, mechanical stiffness and biological response by seeded immortalized human mesenchymal stem cells - adhesion and pore bridging behavior - were investigated. The internal structure of a xyz-gradient scaffold was proven via X-ray based micro computed tomography (µCT). Silicone was used as a model material to investigate the impact of printing velocity and strand distance on the shape fidelity of the sinus pattern for soft matter printing. The impact of different sinus patterns on mechanical properties was assessed. Density and mechanical properties of CPC scaffolds were successfully adjusted without an adverse effect on adhesion and cell number development. In a proof-of-concept experiment, a sinus-adjusted density gradient in an anatomically shaped construct (human vertebral body) defined via clinical computed tomography (CT) data was demonstrated. This fills a technological gap for extrusion-based printing of freely adjustable, continuously guidable infill density gradients in all spatial directions. STATEMENT OF SIGNIFICANCE: 3D extrusion printing of biomaterials allows the generation of anatomically shaped, patient-specific implants or tissue engineering scaffolds. The density of such a structure is typically adjusted by the strand-to-strand distance of parallel, straight-meandered strands in each deposited layer. By printing in a sinusoidal pattern, design of density gradients is possible with a free, spatial resolution in x-, y- and z-direction. We demonstrated that porosity and mechanical properties can be freely adapted in this way without an adverse effect on cell adhesion. With the example of a CT dataset of a human spine, the anisotropic pattern of a vertebral body was resembled by this printing technique that can be translated to various patterns, materials and application.

Multifunctional Three-Dimensional Printed Copper Loaded Calcium Phosphate Scaffolds for Bone Regeneration

Bone regeneration using inorganic nanoparticles is a robust and safe approach. In this paper, copper nanoparticles (Cu NPs) loaded with calcium phosphate scaffolds were studied for their bone regeneration potential in vitro. The pneumatic extrusion method of 3D printing was employed to prepare calcium phosphate cement (CPC) and copper loaded CPC scaffolds with varying wt% of copper nanoparticles. A new aliphatic compound Kollisolv MCT 70 was used to ensure the uniform mixing of copper nanoparticles with CPC matrix. The printed scaffolds were studied for physico-chemical characterization for surface morphology, pore size, wettability, XRD, and FTIR. The copper ion release was studied in phosphate buffer saline at pH 7.4. The in vitro cell culture studies for the scaffolds were performed using human mesenchymal stem cells (hMSCs). The cell proliferation study in CPC-Cu scaffolds showed significant cell growth compared to CPC. The CPC-Cu scaffolds showed improved alkaline phosphatase activity and angiogenic potential compared to CPC. The CPC-Cu scaffolds showed significant concentration dependent antibacterial activity in Staphylococcus aureus. Overall, the CPC scaffolds loaded with 1 wt% Cu NPs showed improved activity compared to other CPC-Cu and CPC scaffolds. The results showed that copper has improved the osteogenic, angiogenic and antibacterial properties of CPC scaffolds, facilitating better bone regeneration in vitro.

Preparation and Properties of Self‐Setting Calcium Phosphate Scaffolds: Effect of Pore Architecture

Self‐setting calcium phosphate cement (CPC) scaffold with interconnected macropores is hard to prepare without compromising its hydration reaction. Herein, an indirect 3D printing method is using to prepare CPC scaffolds. Detailedly, polycaprolactone (PCL)‐sacrificed models with different strut sizes are first printed by 3D plotting technique, and then the CPC pastes are perfused into the PCL models and then self‐setting. After the removal of the PCL models, the CPC scaffolds with different pore sizes are obtained. It is showed in the results that the prepared CPC scaffolds had uniform shape and 3D interconnected macropore structure. Meanwhile, the compressive strength of CPC scaffolds all meets the requirements of repairing cancellous bone. More interestingly, the CPC scaffold with about 300 μm in macropore size is more conducive to the ingrowth and osteogenesis differentiation of mouse bone marrow mesenchymal stem cells. As a whole, the indirect 3D printing method is a promising strategy for preparing CPC scaffolds, which are hard to form directly by 3D printing technique.

Development of Neovasculature in Axially Vascularized Calcium Phosphate Cement Scaffolds

Augmenting the vascular supply to generate new tissues, a crucial aspect in regenerative medicine, has been challenging. Recently, our group showed that calcium phosphate can induce the formation of a functional neo-angiosome without the need for microsurgical arterial anastomosis. This was a preclinical proof of concept for biomaterial-induced luminal sprouting of large-diameter vessels. In this study, we investigated if sprouting was a general response to surgical injury or placement of an inorganic construct around the vessel. Cylindrical biocement scaffolds of differing chemistries were placed around the femoral vein. A contrast agent was used to visualize vessel ingrowth into the scaffolds. Cell populations in the scaffold were mapped using immunohistochemistry. Calcium phosphate scaffolds induced 2.7-3 times greater volume of blood vessels than calcium sulphate or magnesium phosphate scaffolds. Macrophage and vSMC populations were identified that changed spatially and temporally within the scaffold during implantation. NLRP3 inflammasome activation peaked at weeks 2 and 4 and then declined; however, IL-1β expression was sustained over the course of the experiment. IL-8, a promoter of angiogenesis, was also detected, and together, these responses suggest a role of sterile inflammation. Unexpectedly, the effect was distinct from an injury response as a result of surgical placement and also was not simply a foreign body reaction as a result of placing a rigid bioceramic next to a vein, since, while the materials tested had similar microstructures, only the calcium phosphates tested elicited an angiogenic response. This finding then reveals a potential path towards a new strategy for creating better pro-regenerative biomaterials.

In vivo prevascularization strategy enhances neovascularization of β-tricalcium phosphate scaffolds in bone regeneration

BackgroundNeovascularization is critical for bone regeneration. Numerous studies have explored prevascularization preimplant strategies, ranging from calcium phosphate cement (CPC) scaffolds to co-culturing CPCs with stem cells. The aim of the present study was to evaluate an alternative in vivo prevascularization approach, using preimplant-prepared macroporous beta-tricalcium phosphate (β-TCP) scaffolds and subsequent transplantation in bone defect model.MethodsThe morphology of β-TCPs was characterized by scanning electron microscopy. After 3 weeks of prevascularization within a muscle pouch at the lateral size of rat tibia, we transplanted prevascularized macroporous β-TCPs in segmental tibia defects, using blank β-TCPs as a control. Extent of neovascularization was determined by angiography and immunohistochemical (IHC) evaluations. Tibia samples were collected at different time points for biomechanical, radiological, and histological analyses. RT-PCR and western blotting were used to evaluate angio- and osteo-specific markers.ResultsWith macroporous β-TCPs, we documented more vascular and supporting tissue invasion in the macroporous β-TCPs with prior in vivo prevascularization. Radiography, biomechanical, IHC, and histological analyses revealed considerably more vascularity and bone consolidation in β-TCP scaffolds that had undergone the prevascularization step compared to the blank β-TCP scaffolds. Moreover, the prevascularization treatment remarkably upregulated mRNA and protein expression of BMP2 and vascular endothelial growth factor (VEGF) during bone regeneration.ConclusionThis novel in vivo prevascularization strategy successfully accelerated vascular formation to bone regeneration. Our findings indicate that prevascularized tissue-engineered bone grafts have promising potential in clinical applications.The translational potential of this articleThis study indicates a novel in vivo prevascularization strategy for growing vasculature on β-TCP scaffolds to be used for repair of large segmental bone defects, might serve as a promising tissue-engineered bone grafts in the future.

3D-Printed Scaffolds Promote Angiogenesis by Recruiting Antigen-Specific T Cells

The immune response after implantation is a primary determinant of the tissue-repair effects of three-dimensional (3D)-printed scaffolds. Thus, scaffolds that can subtly regulate immune responses may display extraordinary functions. Inspired by the angiogenesis promotion effect of humoral immune response, we covalently combined mesoporous silica microrod (MSR)/polyethyleneimine (PEI)/ovalbumin (OVA) self-assembled vaccines with 3D-printed calcium phosphate cement (CPC) scaffolds for local antigen-specific immune response activation. With the response activated, antigen-specific CD4 + T helper 2 (Th2) cells can be recruited to promote early angiogenesis. The silicon (Si) ions from MSRs can accelerate osteogenesis, with an adequate blood supply being provided. At room temperature, scaffolds with uniformly interconnected macropores were printed using a self-setting CPC-based printing paste, which promoted the uniform dispersion and structural preservation of functional polysaccharides oxidized hyaluronic acid (OHA) inside. Sustained release of OVA was achieved with MSR/PEI covalently attached to scaffolds rich in aldehyde groups as the vaccine carrier. The vaccine-loaded scaffolds effectively recruited and activated dendritic cells (DCs) for antigen presentation and promoted the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in vitro . When embedded subcutaneously in vivo , the vaccine-loaded scaffolds increased the proportion of Th2 cells in the spleen and locally recruited antigen-specific T cells to promote angiogenesis in and around the scaffold. Furthermore, the result in a rat skull defect-repair model indicated that the antigen-specific vaccine-loaded scaffolds promoted the regeneration of vascularized bone. This method may provide a novel concept for patient-specific implant design for angiogenesis promotion.

3D-printed Mechanically Strong Calcium Phosphate Cement Scaffold with Metformin/Stem Cell-encapsulating Alginate Microbeads for Bone Tissue Engineering

3D-printed Mechanically Strong Calcium Phosphate Cement Scaffold with Metformin/Stem Cell-encapsulating Alginate Microbeads for Bone Tissue Engineering

Fabrication of Radio-Opaque and Macroporous Injectable Calcium Phosphate Cement.

The aim of this work was the development of injectable radio-opaque and macroporous calcium phosphate cement (CPC) to be used as a bone substitute for the treatment of pathologic vertebral fractures. A CPC was first rendered radio-opaque by the incorporation of zirconium dioxide (ZrO2). In order to create macroporosity, poly lactic-co-glycolic acid (PLGA) microspheres around 100 μm were homogeneously incorporated into the CPC as observed by scanning electron microscopy. Physicochemical analyses by X-ray diffraction and Fourier transform infrared spectroscopy confirmed the brushite phase of the cement. The mechanical properties of the CPC/PLGA cement containing 30% PLGA (wt/wt) were characterized by a compressive strength of 2 MPa and a Young's modulus of 1 GPa. The CPC/PLGA exhibited initial and final setting times of 7 and 12 min, respectively. Although the incorporation of PLGA microspheres increased the force necessary to inject the cement and decreased the percentage of injected mass as a function of time, the CPC/PLGA appeared fully injectable at 4 min. Moreover, in comparison with CPC, CPC/PLGA showed a full degradation in 6 weeks (with 100% mass loss), and this was associated with an acidification of the medium containing the CPC/PLGA sample (pH of 3.5 after 6 weeks). A cell viability test validated CPC/PLGA biocompatibility, and in vivo analyses using a bone defect assay in the caudal vertebrae of Wistar rats showed the good opacity of the CPC through the tail and a significant increased degradation of the CPC/PLGA cement a month after implantation. In conclusion, this injectable CPC scaffold appears to be an interesting material for bone substitution.

Human Periodontal Ligament Stem Cell and Umbilical Vein Endothelial Cell Co-Culture to Prevascularize Scaffolds for Angiogenic and Osteogenic Tissue Engineering.

(1) Background: Vascularization remains a critical challenge in bone tissue engineering. The objective of this study was to prevascularize calcium phosphate cement (CPC) scaffold by co-culturing human periodontal ligament stem cells (hPDLSCs) and human umbilical vein endothelial cells (hUVECs) for the first time; (2) Methods: hPDLSCs and/or hUVECs were seeded on CPC scaffolds. Three groups were tested: (i) hUVEC group (hUVECs on CPC); (ii) hPDLSC group (hPDLSCs on CPC); (iii) co-culture group (hPDLSCs + hUVECs on CPC). Osteogenic differentiation, bone mineral synthesis, and microcapillary-like structures were evaluated; (3) Results: Angiogenic gene expressions of co-culture group were 6–9 fold those of monoculture. vWF expression of co-culture group was 3 times lower than hUVEC-monoculture group. Osteogenic expressions of co-culture group were 2–3 folds those of the hPDLSC-monoculture group. ALP activity and bone mineral synthesis of co-culture were much higher than hPDLSC-monoculture group. Co-culture group formed capillary-like structures at 14–21 days. Vessel length and junction numbers increased with time; (4) Conclusions: The hUVECs + hPDLSCs co-culture on CPC scaffold achieved excellent osteogenic and angiogenic capability in vitro for the first time, generating prevascularized networks. The hPDLSCs + hUVECs co-culture had much better osteogenesis and angiogenesis than monoculture. CPC scaffolds prevacularized via hPDLSCs + hUVECs are promising for dental, craniofacial, and orthopedic applications.

Regenerating Craniofacial Dental Defects With Calcium Phosphate Cement Scaffolds: Current Status and Innovative Scope Review

The management and treatment of dental and craniofacial injuries have continued to evolve throughout the last several decades. Limitations with autograft, allograft, and synthetics created the need for more advanced approaches in tissue engineering. Calcium phosphate cements (CPC) are frequently used to repair bone defects. Since their discovery in the 1980s, extensive research has been conducted to improve their properties, and emerging evidence supports their increased application in bone tissue engineering. This review focuses on the up-to-date performance of calcium phosphate cement (CPC) scaffolds and upcoming promising dental and craniofacial bone regeneration strategies. First, we summarized the barriers encountered in CPC scaffold development. Second, we compiled the most up to date in vitro and in vivo literature. Then, we conducted a systematic search of scientific articles in MEDLINE and EMBASE to screen the related studies. Lastly, we revealed the current developments to effectively design CPC scaffolds and track the enhanced viability and therapeutic efficacy to overcome the current limitations and upcoming perspectives. Finally, we presented a timely and opportune review article focusing on the significant potential of CPC scaffolds for dental and craniofacial bone regeneration, which will be discussed thoroughly. CPC offers multiple capabilities that may be considered toward the oral defects, expecting a future outlook in nanotechnology design and performance.