Development and characterization of waste equine bone-derived calcium phosphate cements with human alveolar bone-derived mesenchymal stem cells.

Calcium phosphate cements (CPCs) are regarded as promising graft substitutes for bone tissue engineering. However, their wide use is limited by the high cost associated with the complex synthetic processes involved in their fabrication. Cheaper xenogeneic calcium phosphate (CaP) materials derived from waste animal bone may solve this problem. Moreover, the surface topography, mechanical strength, and cellular function of CPCs are influenced by the ratio of micro- to nano-sized CaP (M/NCaP) particles. In this study, we developed waste equine bone (EB)-derived CPCs with various M/NCaP particle ratios to examine the potential capacity of EB-CPCs for bone grafting materials. Our study showed that increasing the number of NCaP particles resulted in reductions in roughness and porosity while promoting smoother surfaces of EB-CPCs. Changes in the chemical properties of EB-CPCs by NCaP particles were observed using X-ray diffractometry. The mechanical properties and cohesiveness of the EB-CPCs improved as the NCaP particle content increased. In an in vitro study, EB-CPCs with a greater proportion of MCaP particles showed higher cell adhesion. Alkaline phosphatase activity indicated that osteogenic differentiation by EB-CPCs was promoted with increased NCaP particle content. These results could provide a design criterion for bone substitutes for orthopedic disease, including periodontal bone defects.

Long-Term Retarded Release for the Proteasome Inhibitor Bortezomib through Temperature-Sensitive Dendritic Glycopolymers as Drug Delivery System from Calcium Phosphate Bone Cement.

For the local treatment of bone defects, highly adaptable macromolecular architectures are still required as drug delivery system (DDS) in solid bone substitute materials. Novel DDS fabricated by host-guest interactions between ß-cyclodextrin-modified dendritic glycopolymers and adamantane-modified temperature-sensitive polymers for the proteasome inhibitor bortezomib (BZM) is presented. These DDS induce a short- and long-term (up to two weeks) retarded release of BZM from calcium phosphate bone cement (CPC) in comparison to a burst release of the drug alone. Different release parameters of BZM/DDS/CPC are evaluated in phosphate buffer at 37 °C to further improve the long-term retarded release of BZM. This is achieved by increasing the amount of drug (50-100 µg) and/or DDS (100-400 µg) versus CPC (1 g), by adapting the complexes better to the porous bone cement environment, and by applying molar ratios of excess BZM toward DDS with 1:10, 1:25, and 1:100. The temperature-sensitive polymer shells of BZM/DDS complexes in CPC, which allow drug loading at room temperature but are collapsed at body temperature, support the retarding long-term release of BZM from DDS/CPC. Thus, the concept of temperature-sensitive DDS for BZM/DDS complexes in CPC works and matches key points for a local therapy of osteolytic bone lesions.

Effect of the Addition of Alginate and/or Tetracycline on Brushite Cement Properties.

Calcium phosphate cements have the advantage that they can be prepared as a paste that sets in a few minutes and can be easily adapted to the shape of the bone defect, which facilitates its clinical application. In this research, six formulations of brushite (dicalcium phosphate dihydrated) cement were obtained and the effect of the addition of sodium alginate was analyzed, such as its capacity as a tetracycline release system. The samples that contain sodium alginate set in 4 or 5 min and showed a high percentage of injectability (93%). The cements exhibit compression resistance values between 1.6 and 2.6 MPa. The drug was released in a range between 12.6 and 13.2% after 7 days. The antimicrobial activity of all the cements containing antibiotics was proven. All samples reached values of cell viability above 70 percent. We also observed that the addition of the sodium alginate and tetracycline improved the cell viability.

Comparison of complications in cranioplasty with various materials: a systematic review and meta-analysis.

Objective: Meta-analysis to evaluate complications in the use of autogenous bone and bone substitutes and to compare bone substitutes, specifically HA, polyetheretherketone (PEEK) and titanium materials.Methods: Search of PubMed, Cochrane, Embase and Google scholar to identify all citations from 2010 to 2019 reporting complications regarding materials used in cranioplasty.Results: 20 of 2266 articles met the inclusion criteria, including a total of 2913 patients. The odds of overall complication were significantly higher in the autogenous bone group (n = 214/644 procedures, 33.2%) than the bone substitute groups (n = 116/436 procedures, 26.7%, CI 1.29-2.35, p < 0.05). In bone substitutes groups, there was no significant difference in overall complication rate between HA and Ti (OR, 1.2; 95% CI, 0.47-3.14, p = 0.69). PEEK has lower overall complication rates (OR, 0.51; 95% CI, 0.30-0.87, p = 0.01) and lower implant exposure rates (OR, 0.17; 95% CI, 0.06-0.53, p = 0.002) than Ti, but there was no significant difference in infection rates and postoperative hematoma rates.Conclusions: Cranioplasty is associated with high overall complication rates with the use of autologous bone grafts compared with bone substitutes. PEEK has a relatively low overall complication rates in substitutes groups, but still high infection rates and postoperative hematoma rates. Thus, autologous bone grafts should only be used selectively, and prospective long-term studies are needed to further refine a better material in cranioplasty.

[Mechanical Reinforcement Strategy of Calcium Phosphate Cements by Loading Polymers].

Calcium phosphate cement (CPC) is well known for the excellent bioactivity and biocompatibility, however, CPC has been used only for the repair of non-load bearing bone defects due to its brittle nature and low flexural strength. Polymer reinforced CPC has been considered as one of the most effective strategies for mechanical reinforcement. This paper summarizes various kinds of polymers loaded CPC:fiber reinforcement, microsphere reinforcement and dual setting cements. It is aimed to analyze the advantages, disadvantages and principles of the polymers reinforced CPC, and so as to lay a foundation for the further research of improving and manufacturing the CPC with ideal mechanical properties.

Calcium silicate/calcium phosphate biphasic cements for vital pulp therapy: chemical-physical properties and human pulp cells response.

OBJECTIVES: The aim was to test the properties of experimental calcium silicate/calcium phosphate biphasic cements with hydraulic properties designed for vital pulp therapy as direct pulp cap and pulpotomy. METHODS: CaSi-αTCP and CaSi-DCDP were tested for ion-releasing ability, solubility, water sorption, porosity, ability to nucleate calcium phosphates, and odontoblastic differentiation­alkaline phosphatase (ALP) and osteocalcin (OCN) upregulation­of primary human dental pulp cells (HDPCs). RESULTS: The materials showed high Ca and OH release, high open pore volume and apparent porosity, and a pronounced ability to nucleate calcium phosphates on their surface. HDPCs treated with CaSi-αTCP showed a strong upregulation of ALP and OCN genes, namely a tenfold increase for OCN and a threefold increase for ALP compared to the control cells. Conversely, CaSi-DCDP induced a pronounced OCN gene upregulation but had no effect on ALP gene regulation. CONCLUSIONS: Both cements showed high biointeractivity (release of Ca and OH ions) correlated with their marked ability to nucleate calcium phosphates. CaSi-αTCP cement proved to be a potent inducer of ALP and OCN genes as characteristic markers of mineralization processes normally poorly expressed by HDPCs. CLINICAL RELEVANCE: Calcium silicate/calcium phosphate cements appear to be attractive new materials for vital pulp therapy as they may provide odontogenic/dentinogenic chemical signals for pulp regeneration and healing, and dentin formation in regenerative endodontics.

Preferentially Biodegradable Gypsum Fibers Endowing Invisible Microporous Structures and Enhancing Osteogenic Capability of Calcium Phosphate Cements.

It is known that hydroxyapatite-type calcium phosphate cement (CPC) shows appreciable self-curing properties, but the phase transformation products often lead to slow biodegradation and disappointing osteogenic responses. Herein, we developed an innovative strategy to endow invisible micropore networks, which could tune the microstructures and biodegradation of α-tricalcium phosphate (α-TCP)-based CPC by gypsum fibers, and the osteogenic capability of the composite cements could be enhanced in vivo. The gypsum fibers were prepared via extruding the gypsum powder/carboxylated chitosan (CC) slurry through a 22G nozzle (410 µm in diameter) and collecting with a calcium salt solution. Then, the CPCs were prepared by mixing the α-TCP powder with gypsum fibers (0-24 wt %) and an aqueous solution to form self-curing cements. The physicochemical characterizations showed that injectability was decreased with an increase in the fiber contents. The µCT reconstruction demonstrated that the gypsum fiber could be distributed in the CPC substrate and produce long-range micropore architectures. In particular, incorporation of gypsum fibers would tune the ion release, produce tunnel-like pore networks in vitro, and promote new bone tissue regeneration in rabbit femoral bone defects in vivo. Appropriate gypsum fibers (16 and 24 wt %) could enhance bone defect repair and cement biodegradation. These results demonstrate that the highly biodegradable cement fibers could mediate the microstructures of conventional CPC biomaterials, and such a bicomponent composite strategy may be beneficial for expanding clinical CPC-based applications.

Incorporating the Antioxidant Fullerenol into Calcium Phosphate Bone Cements Increases Cellular Osteogenesis without Compromising Physical Cement Characteristics.

Herein, fullerenol (Ful), a highly water-soluble derivative of C60 fullerene with demonstrated antioxidant activity, is incorporated into calcium phosphate cements (CPCs) to enhance their osteogenic ability. CPCs with added carboxymethyl cellulose/gelatin (CMC/Gel) are doped with biocompatible Ful particles at concentrations of 0.02, 0.04, and 0.1 wt v%-1 and evaluated for Ful-mediated mechanical performance, antioxidant activity, and in vitro cellular osteogenesis. CMC/gel cements with the highest Ful concentration decrease setting times due to increased hydrogen bonding from Ful's hydroxyl groups. In vitro studies of reactive oxygen species (ROS) scavenging with CMC/gel cements demonstrate potent antioxidant activity with Ful incorporation and cement scavenging capacity is highest for 0.02 and 0.04 wt v%-1 Ful. In vitro cytotoxicity studies reveal that 0.02 and 0.04 wt v%-1 Ful cements also protect cellular viability. Finally, increase of alkaline phosphatase (ALP) activity and expression of runt-related transcription factor 2 (Runx2) in MC3T3-E1 pre-osteoblast cells treated with low-dose Ful cements demonstrate Ful-mediated osteogenic differentiation. These results strongly indicate that the osteogenic abilities of Ful-loaded cements are correlated with their antioxidant activity levels. Overall, this study demonstrates exciting potential of Fullerenol as an antioxidant and proosteogenic additive for improving the performance of calcium phosphate cements in bone reconstruction procedures.

Calcium phosphate bone cements as local drug delivery systems for bone cancer treatment.

Bone cancer is usually a metastatic disease, affecting people of all ages. Its effective therapy requires a targeted drug administration locally at the cancer site so that the surrounding healthy organs and tissues stay unharmed. Upon a thorough literature search, a tremendous number of published articles are reporting on development of calcium phosphate cements (CPCs) for the treatment of a variety of diseases, such as osteoporosis, osteoarthritis, osteomyelitis, and other musculoskeletal disorders. However, just a limited number of research employs CPCs specifically for bone cancer treatment. In this review article, we study the factors influencing the local drug release from CPCs and particularly focus on bone cancer therapy. Finally, we locate the deficiencies in the literature regarding this specific topic and propose which other perspectives should be considered and discussed in future articles.

Sudoku of porous, injectable calcium phosphate cements - Path to osteoinductivity.

With the increase of global population, people's life expectancy is growing as well. Humans tend to live more active lifestyles and, therefore, trauma generated large defects become more common. Instances of tumour resection or pathological conditions and complex orthopaedic issues occur more frequently increasing necessity for bone substitutes. Composition of calcium phosphate cements (CPCs) is comparable to the chemical structure of bone minerals. Their ability to self-set and resorb in vivo secures a variety of potential applications in bone regeneration. Despite the years-long research and several products already reaching the market, finding the right properties for calcium phosphate cement to be osteoinductive and both injectable and suitable for clinical use is still a sudoku. This article is focused on injectable, porous CPCs, reviewing the latest developments on the path toward finding osteoinductive material, which is suitable for injection.

Application of a Mytilus edulis-derived promoting calcium absorption peptide in calcium phosphate cements for bone.

The IEELEEELEAER peptide (PIE) identified from the protein hydrolysate of Mytilus edulis is reported to enhance osteoblast growth and differentiation, which also possesses a superior bone formation ability both in vitro and in vivo. Moreover, PIE bound to calcium spontaneously at the stoichiometry of 1:1, and there were amino nitrogen and carboxyl oxygen atoms in 2 glutamic acid residues at the calcium-binding sites in the PIE. The PIE-calcium complex facilitated calcium uptake through the Caco-2 cell monolayers. Incorporation of PIE into calcium phosphate cements enhanced calcium ion uptake and proliferation of osteoblasts and inhibit bacteria. This study suggest that calcium phosphate cements supplemented with PIE can serve as a potentially efficient material for bone graft used during spinal surgery.

α-TCP-based calcium phosphate cements: A critical review.

Calcium phosphates are promising materials for applications in bone repair and substitution, particularly for their bioactivity and ability to form self-setting cements. Among them, α-tricalcium phosphate (α-TCP) stands out due to its high solubility, its hydration reaction and bioresorbability. The synthesis of α-TCP is particularly complex and the interactions between some of the synthesis parameters are still not completely understood. The variety of methods available to synthesize α-TCP has provided a substantial variance in the properties of α-TCP-based cements and the decision about which method, parameters and starting reagents will be used for the powder's synthesis is determinant of the properties of the resulting material. Therefore, this review paper focuses on α-TCP's synthesis and properties, presenting the synthesis methods currently in use as well as a discussion of how the synthesis parameters and the cement preparation affect the reactivity and mechanical properties of the material, providing a guide for the selection of the most suitable process for each α-TCP application. STATEMENT OF SIGNIFICANCE: α-TCP is a calcium phosphate and it is currently one of the most investigated bioceramics for applications that explore its bioresorbability and the hydration reaction of α-TCP-based cements. Despite the increasing number of publications on the topic, there are still aspects not well understood. This review article aims at contributing to this fascinating subject by offering an update on the state of the art of α-TCP's synthesis methods, while also addressing topics that are not often discussed about this material, such as the preparation of α-TCP-based cements and how its parameters affect the properties of the resulting cements.

Factors influencing the drug release from calcium phosphate cements.

Thanks to their biocompatibility, biodegradability, injectability and self-setting properties, calcium phosphate cements (CPCs) have been the most economical and effective biomaterials of choice for use as bone void fillers. They have also been extensively used as drug delivery carriers owing to their ability to provide for a steady release of various organic molecules aiding the regeneration of defective bone, including primarily antibiotics and growth factors. This review provides a systematic compilation of studies that reported on the controlled release of drugs from CPCs in the last 25 years. The chemical, compositional and microstructural characteristics of these systems through which the control of the release rates and mechanisms could be achieved have been discussed. In doing so, the effects of (i) the chemistry of the matrix, (ii) porosity, (iii) additives, (iv) drug types, (v) drug concentrations, (vi) drug loading methods and (vii) release media have been distinguished and discussed individually. Kinetic specificities of in vivo release of drugs from CPCs have been reviewed, too. Understanding the kinetic and mechanistic correlations between the CPC properties and the drug release is a prerequisite for the design of bone void fillers with drug release profiles precisely tailored to the application area and the clinical picture. The goal of this review has been to shed light on these fundamental correlations.

Role of nanostructured materials in hard tissue engineering.

The rise in the use of biomaterials in bone regeneration in the last decade has exponentially multiplied the number of publications, methods, and approaches to improve and optimize their functionalities and applications. In particular, biomimetic strategies based on the self-assembly of molecules to design, create and characterize nanostructured materials have played a very relevant role. We address this idea on four different but related points: self-setting bone cements based on calcium phosphate, as stable tissue support and regeneration induction; metallic prosthesis coatings for cell adhesion optimization and prevention of inflammatory response exacerbation; bio-adhesive hybrid materials as multiple drug delivery localized platforms and finally bio-inks. The effect of the physical, chemical, and biological properties of the newest biomedical devices on their bone tissue regenerative capacity are summarized, described, and analyzed in detail. The roles of experimental conditions, characterization methods and synthesis routes are emphasized. Finally, the future opportunities and challenges of nanostructured biomaterials with their advantages and shortcomings are proposed in order to forecast the future directions of this field of research.

The strategy of composite grafting with BMP2-Loaded calcium phosphate cements and autogenous bone for alveolar cleft reconstruction.

Purpose: To remedy the drawbacks of traditional autogenous bone harvesting in alveolar bone grafting (ABG), a novel strategy of composite grafting with BMP2-loaded calcium phosphate cements (BMP2-CPC) and autogenous bone harvested by minimally invasive technique was developed and evaluated for its bone-repairing efficacy. Materials and methods: A chart review was conducted for 19 patients with unilateral alveolar clefts who underwent secondary ABG from 2017 to 2020. Of the enrolled patients, 9 patients underwent grafting with autogenous bone harvested by traditional trap door technique (group I), and 10 patients underwent grafting with the composite graft comprising BMP2-CPC and autogenous bone harvested by minimally invasive technique at a ratio of 1:1 by volume (group II). The clinical performance of the composite graft was comprehensively evaluated in terms of clinical, radiographic and histological perspectives. Results: The present results demonstrated that the composite graft exhibited satisfactory bone-repairing efficacy comparable to that of the autogenous bone graft on the premise of lower amount of harvested bone. The post-surgical resorption of bone volume and vertical height of grafted area was significantly slower in group II. The favourable resorption performance of BMP2-CPC contributed to preserving the post-surgical bony contour reconstructed with the composite graft. Conclusion: The composite graft comprising BMP2-CPC and autogenous bone harvested by minimally invasive technique was demonstrated to be an eligible alternative for application in ABG, especially for its improved resorption performance in preserving post-surgical bony contour.

The osteogenesis effect of rhBMP2-loaded calcium phosphate cements in repairing dental extraction sockets.

PURPOSE: After extracting impacted mandibular third molars (IMM3), the resulting bone loss at the distal surface of the distal root of mandibular second molars (MM2) is responsible for the poor stability of MM2. This study aimed to identify the clinical osteogenesis effect of recombinant human bone morphogenetic protein-2 (rhBMP-2)-loaded calcium phosphate cements (CPCs) and rhBMP-2 delivery systems (rhBMP-2/CPCs, named CPCII) on bone loss repair at the distal surface of the MM2 distal root after IMM3 extraction. METHODS: Written informed consent was obtained from every participant whose IMM3 needed extraction. The impact of IMM3 on both sides was basically identical. From April 2014 to March 2016, extraction of IMM3 was performed in 9 patients (5 males/4 females, 26-42 years old). One side was randomly selected as the experimental group, and CPCII systems were implanted into the distal surface of the distal root in dental extraction sockets. The wounds on the other side were sutured and allowed to heal naturally (be treated as the control group). New bone formation in the alveolar fossa was detected 3 and 12 months after the operation by cone-beam computed tomography (CBCT) to measure the distance from the cementoenamel junction (CEJ) to the crest of the alveolar ridge (CAR). RESULTS: The CAR-CEJ distance on the test side was less than that on the control side (P<0.5). CONCLUSION: The quantity of new bone formation in the experimental group was greater than that in the control group. CPCII systems have osteogenic potential in the healing process of tooth extraction sockets.

Calcium phosphate cements: Optimization toward biodegradability.

Synthetic calcium phosphate (CaP) ceramics represent the most widely used biomaterials for bone regenerative treatments due to their biological performance that is characterized by bioactivity and osteoconductive properties. From a clinical perspective, injectable CaP cements (CPCs) are highly appealing, as CPCs can be applied using minimally invasive surgery and can be molded to optimally fill irregular bone defects. Such CPCs are prepared from a powder and a liquid component, which upon mixing form a paste that can be injected into a bone defect and hardens in situ within an appropriate clinical time window. However, a major drawback of CPCs is their poor degradability. Ideally, CPCs should degrade at a suitable pace to allow for concomitant new bone to form. To overcome this shortcoming, control over CPC degradation has been explored using multiple approaches that introduce macroporosity within CPCs. This strategy enables faster degradation of CPC by increasing the surface area available to interact with the biological surroundings, leading to accelerated new bone formation. For a comprehensive overview of the path to degradable CPCs, this review presents the experimental procedures followed for their development with specific emphasis on (bio)material properties and biological performance in pre-clinical bone defect models.

Calcium phosphate cement reinforced with poly (vinyl alcohol) fibers: An experimental and numerical failure analysis.

Calcium phosphate cements (CPCs) have been widely used during the past decades as biocompatible bone substitution in maxillofacial, oral and orthopedic surgery. CPCs are injectable and are chemically resemblant to the mineral phase of native bone. Nevertheless, their low fracture toughness and high brittleness reduce their clinical applicability to weakly loaded bones. Reinforcement of CPC matrix with polymeric fibers can overcome these mechanical drawbacks and significantly enhance their toughness and strength. Such fiber-reinforced calcium phosphate cements (FRCPCs) have the potential to act as advanced bone substitute in load-bearing anatomical sites. This work achieves integrated experimental and numerical characterization of the mechanical properties of FRCPCs under bending and tensile loading. To this end, a 3-D numerical gradient enhanced damage model combined with a dimensionally-reduced fiber model are employed to develop a computational model for material characterization and to simulate the failure process of fiber-reinforced CPC matrix based on experimental data. In addition, an advanced interfacial constitutive law, derived from micromechanical pull-out tests, is used to represent the interaction between the polymeric fiber and CPC matrix. The presented computational model is successfully validated with the experimental results and offers a firm basis for further investigations on the development of numerical and experimental analysis of fiber-reinforced bone cements.

Antibacterial calcium phosphate composite cements reinforced with silver-doped magnesium phosphate (newberyite) micro-platelets.

This article demonstrates our efforts in developing and evaluating all-ceramic, biodegradable composites of calcium phosphate cements (CPCs) reinforced with silver (Ag)-doped magnesium phosphate (MgP) crystals. Two primary goals of this study were to 1) enhance CPC's poor mechanical properties with micro-platelet reinforcement, and 2) impart antibacterial functionalities in composites with the aim to inhibit surgical site infections (SSI). The work embodies three novel features. First, as opposed to well-known reinforcements with whisker or fiber-like morphology, we explored micro-platelets for the first time as the strengthening phase in the CPC matrix. Second, in contrast to conventional polymeric or calcium phosphate (CaP) fibrous reinforcements, newberyite (NB, MgHPO4.3H2O) micro-platelets belonging to the less explored yet promising MgP family, were evaluated as reinforcements for the first time. Third, NB micro-platelets were doped with Ag+ ions (AgNB, Ag content: 2 wt%) for enhancing antibacterial functionalities. Results indicated that 1 wt% of AgNB micro-platelet incorporation in the CPC matrix enhanced the compressive and flexural strengths by 200% and 140% respectively as compared to the un-reinforced ones. Besides, antibacterial assays revealed effective bactericidal functionalities (>99% bacteria kill) of the AgNB reinforced CPCs against Escherichia coli. Finally, cytocompatibility studies confirmed favorable pre-osteoblast cell proliferation and differentiation in vitro. Hence, this effort was successful in developing a self-setting and injectable AgNB reinforced CPC composition with favorable mechanical and antibacterial properties.

Experimental and numerical analysis on bending and tensile failure behavior of calcium phosphate cements.

Since their discovery in the 1980s, injectable self-setting calcium phosphate cements (CPCs) are frequently used in orthopedic, oral and maxillofacial surgery due to their chemical resemblance to the mineral phase of native bone. However, these cements are very brittle, which complicates their application in load-bearing anatomical sites. Polymeric fibers can be used to transform brittle calcium phosphate cements into ductile and load-bearing biomaterials. To understand and optimize this process of fiber reinforcement, it is essential to characterize the mechanical properties of fiber-free calcium phosphate matrices in full detail. However, the mechanical performance of calcium phosphate cements is usually tested under compression only, whereas bending and tensile tests are hardly performed due to technical limitations. In addition, computational models describing failure behavior of calcium phosphate cements under these clinically more relevant loading scenarios have not yet been developed. Here, we investigate the failure behavior of calcium phosphate cements under bending and tensile loading by combining, for the first time, experimental tests and numerical modeling. To this end, a 3-D gradient-enhanced damage model is developed in a finite element framework, and numerical results are correlated to experimental three-point bending and tensile tests to characterize the mechanical properties of calcium phosphate cements in full detail. The presented computational model is successfully validated against experimental results and is able to predict the mechanical response of calcium phosphate cement under different types of loading with a unique set of parameters. This model offers a solid basis for further experimental and computational studies on the development of load-bearing bone cements.