Macroporous Calcium Phosphate Cement Research Articles

Microstructure and mechanical properties of calcium phosphate cement/gelatine composite scaffold with oriented pore structure for bone tissue engineering

The macroporous calcium phosphate(CPC) cement with oriented pore structure was prepared by freeze casting. SEM observation showed that the macropores in the porous calcium phosphate cement were interconnected aligned along the ice growth direction. The porosity of the as-prepared porous CPC was measured to be 87.6% by Archimede’s principle. XRD patterns of specimens showed that poorly crystallized hydroxyapatite was the main phase present in the hydrated porous calcium phosphate cement. To improve the mechanical properties of the CPC scaffold, the 15% gelatine solution was infiltrated into the pores under vacuum and then the samples were freeze dried to form the CPC/gelatine composite scaffolds. After reinforced with gelatine, the compressive strength of CPC/gelatine composite increased to 5.12 MPa, around fifty times greater than that of the unreinforced macroporous CPC scaffold, which was only 0.1 MPa. And the toughness of the scaffold has been greatly improved via the gelatine reinforcement with a much greater fracture strain. SEM examination of the specimens indicated good bonding between the cement and gelatine. Participating the external load by the deformable gelatine, patching the defects of the CPC pores wall, and crack deflection were supposed to be the reinforcement mechanisms. In conclusion, the calcium phosphate cement/gelatine composite with oriented pore structure prepared in this work might be a potential scaffold for bone tissue engineering.

Correlation between degradation and compressive strength of an injectable macroporous calcium phosphate cement

Variation of compressive strength of a macroporous calcium phosphate cement (CPC) was investigated in the present study. Poly (lactide-co-glycolide acid) (PLGA) microspheres was incorporated with the CPC powder. Sodium citrate solution was used as the cement liquid phase. The formation of macropores and the decline in mechanical strength of the paste was investigated. The results showed that the macroporous CPC exhibited a good property of injectability but the self-setting time was slightly prolonged. With the degradation of PLGA microspheres, the weight loss increased and the average values of compressive strength of the paste was reduced dramatically. The formulation of negative correlation between compressive and weight loss was established as following: y2=−0.794y1+38.278. In conclusion, the present macroporous CPC showed that excellent injectability accompanied with the prolonged setting time, the degradation of microspheres and compressive strength of CPC are also in negative correlation. Regulate the acceleration of degeneration to a proper extents perhaps is another method to enhance the compressive strength in macroporous CPCs.

Gas-Foaming Calcium Phosphate Cement Scaffold Encapsulating Human Umbilical Cord Stem Cells

Tissue engineering approaches are promising to meet the increasing need for bone regeneration. Calcium phosphate cement (CPC) can be injected and self-set to form a scaffold with excellent osteoconductivity. The objectives of this study were to develop a macroporous CPC-chitosan-fiber construct containing alginate-fibrin microbeads encapsulating human umbilical cord mesenchymal stem cells (hUCMSCs) and to investigate hUCMSC release from the degrading microbeads and proliferation inside the porous CPC construct. The hUCMSC-encapsulated microbeads were completely wrapped inside the CPC paste, with the gas-foaming porogen creating macropores in CPC to provide for access to culture media. Increasing the porogen content in CPC significantly increased the cell viability, from 49% of live cells in CPC with 0% porogen to 86% of live cells in CPC with 15% porogen. The alginate-fibrin microbeads started to degrade and release the cells inside CPC at 7 days. The released cells started to proliferate inside the macroporous CPC construct. The live cell number inside CPC increased from 270 cells/mm(2) at 1 day to 350 cells/mm(2) at 21 days. The pore volume fraction of CPC increased from 46.8% to 78.4% using the gas-foaming method, with macropore sizes of approximately 100 to 400 μm. The strength of the CPC-chitosan-fiber scaffold at 15% porogen was 3.8 MPa, which approximated the reported 3.5 MPa for cancellous bone. In conclusion, a novel gas-foaming macroporous CPC construct containing degradable alginate-fibrin microbeads was developed that encapsulated hUCMSCs. The cells had good viability while wrapped inside the porous CPC construct. The degradable microbeads in CPC quickly released the cells, which proliferated over time inside the porous CPC. Self-setting, strong CPC with alginate-fibrin microbeads for stem cell delivery is promising for bone tissue engineering applications.

Preparation and characterisation of porous calcium phosphate bone cement as antibiotic carrier

This study investigated the release of cephalexin monohydrate (CMH) antibitic from macroporous calcium phosphate cement (CPC) over 0·5–300 h in simulated body fluid. Macroporosity was introduced into the cement matrix by using sodium dodecyl sulphate molecules as air entraining agents. The effect of both surfactant and CMH on basic properties of the CPC has been also elaborated. The results showed that the release rate of the drug from a porous CPC was higher than that of the non-porous CPC; however the same release patterns observed for both morphologies indicated a time dependent controlled release. Incorporation of CMH into the cement composition increased the setting time, while the crystallinity of the formed apatite decreased and injectability of the paste improved. In addition, the rate of hydraulic reactions, leading to conversion of the reactants into apatite phase, did not influenced by incorporating both CMH and surfactant into the cement.

Periostal Reconstruction Using New Porcine Microstructured Collagen Membrane and Calcium Phosphate Cement: A Dog Model

The aim of this study was to evaluate the capacity of the association of calcium phosphate cement with collagen membrane for large segmental bone defect. Six adult beagle dog underwent defect creation in the ulna and was reconstructed with combination of macroporous calcium phosphate cement MCPC® and resorbable collagen membrane EZ Cure®. After 6 months of implantation, the samples were analyzed with Micro CT, light microscopy and SEM using BSE. Bridging of the defect with a lamellar and well organized bone was achieved in all animals. MCPC granules resorption was increased at the extremities of the implant. Collagen membrane at the expense of the implant was replaced by periosteum-like formation. The results demonstrate the ability of the composite to reconstruct large segmental and critical size defect in long bone.

Macroporous Calcium Phosphate Cement (MCPC<sup>®</sup>) for Bone Reconstruction in Cranioplasty: Animal Study Performance

We have developed a novel macroporous calcium phosphate cement MCPC® that sets to poorly crystalline apatite after mixing the powder component with an aqueous solution and has interconnective macroporosity We performed cranioplasty on rat model by injecting the new macroporous calcium phosphate cement MCPC®. The mechanical property of the cement is about 12MPa after 24 hours (compression test). The cement matrix is totally transformed into poorly crystalline apatite in 48 hours. This study demonstrates that MCPC® cement was suitable and efficient for parietal bone reconstruction. Its injectability and moldability allows to be used in bone reconstruction surgery and its mechanical properties are compatible to support calvarial reconstruction. In addition, a bone ingrowth onto the BCP granules occurred on time.

PLGA microsphere/calcium phosphate cement composites for tissue engineering: in vitro release and degradation characteristics

Bone cements with biodegradable poly(lactic-co-glycolic acid) (PLGA) microspheres have already been proven to provide a macroporous calcium phosphate cement (CPC) during in situ microsphere degradation. Furthermore, in vitro/in vivo release studies with these PLGA microsphere/CPC composites (PLGA/CPCs) showed a sustained release of osteo-inductive growth factor when drug was distributed inside/onto the microspheres. The goal of this study was to elucidate the mechanism behind drug release from PLGA/CPC. For this, in vitro release and degradation characteristics of a low-molecular-weight PLGA/CPC (M w = 5 kg/mol) were determined using bovine serum albumin (BSA) as a model protein. Two loading mechanisms were applied; BSA was either adsorbed onto the microspheres or incorporated inside the microspheres during double-emulsion. BSA release from PLGA microspheres and CPC was also measured and used as reference. Results show fast degrading polymer microspheres which produced a macroporous scaffold within 4 weeks, but also showed a concomitant release of acidic degradation products. BSA release from the PLGA/CPC was similar to the CPC samples and showed a pattern consisting of a small initial release, followed by a period of almost no sustained release. Separate PLGA microspheres exhibited a high burst release and release efficiency that was higher with the adsorbed samples. Combining degradation and release data we can conclude that for the PLGA/CPC samples BSA re-adsorbed to the cement surface after being released from the microspheres, which was mediated by the pH decrease during microsphere degradation.

Vertebroplasty in Goat Model Using Macroporous Calcium Phosphate Cement (MCPC<sup>®</sup>)

We performed vertebroplasty on goat model by injecting a new macroporous calcium phosphate cement MCPC®. The mechanical property of the cement is about 12MPa after 24 hours (compression test). The cement matrix is totally transformed into poorly crystallized apatite in 48 hours. This study demonstrates that MCPC cement was suitable and efficient for a spine application. Its injectability allows to be used in mini invasive surgery and its mechanical properties are compatible to support spine strength. In addition, a bone ingrowth onto the BCP granules occurred with time.

PL DLLA Calcium Phosphate Composite Combined with Macroporous Calcium Phosphate Cement MCPC® for New Surgical Technologies Combining Resorbable Osteosynthesis and Injectable Bone Substitute

Resorbable osteosynthesis based on PLLA and derivatives will be associated to bone substitute for bone reconstruction. We have performed rand evaluated a composite combining PL DLLA and Biphasic calcium phosphate able to have a), a better controlled hydrolysis in the purpose to preserve on time the mechanical property, and b), for long term efficiency, bone ingrowth at the expense of the osteosynthesis and the associated bone substitute. A new calcium phosphate cement MCPC® was tested with such composite. The novel macroporous calcium phosphate cement MCPC sets to poorly crystalline apatite after mixing the powder component and an aqueous solution. Interconnective macroporosity was induced on time by resorption of one part of the MCPC®. The multiphasic calcium phosphate components in the cement, are resorbed at different rates allowing the replacement by newly formed bone. This study reports the biocompatibility and the interactions of a composite using PL DLLA (Poly [L-Lactide-co-D,L-Lactide] acid) charged with biphasic calcium phosphate granules and a self setting calcium phosphate cement of new generation.

Preparation of Calcium Phosphate Cement Tissue Engineering Scaffold Reinforced with Chitin Fiber

Macroporous calcium phosphate cement (CPC) scaffold was prepared by a salt-leaching method using water-dissoluble rod-shaped monosodium glutamate crystals as poregen.The relationship between the porosity and the amount of poregen was investigated.The results indicate that the porosity of CPC scaffold come up to (79.8±2.3)% with a pore size range of 100-600μm.Chitin fibers was incorporated to reinforce CPC scaffold.The compressive strength of the Chitin fibers reinforced CPC scaffold increases 3 to 4 times higher than that of the CPC scaffold without fibers and the strain of fracture increases remarkably.The scaffold material may be applied in the bone tissue engineering for repair of non-load bearing bone defects.

Strong, macroporous, and in situ-setting calcium phosphate cement-layered structures

Calcium phosphate cement (CPC) is highly promising for clinical uses due to its in situ-setting ability, excellent osteoconductivity and bone-replacement capability. However, the low strength limits its use to non-load-bearing applications. The objectives of this study were to develop a layered CPC structure by combining a macroporous CPC layer with a strong CPC layer, and to investigate the effects of porosity and layer thickness ratios. The rationale was for the macroporous layer to accept tissue ingrowth, while the fiber-reinforced strong layer would provide the needed early-strength. A biopolymer chitosan was incorporated to strengthen both layers. Flexural strength, S (mean±sd; n=6) of CPC-scaffold decreased from (9.7±1.2) to (1.8±0.3) MPa ( p<0.05), when the porosity increased from 44.6% to 66.2%. However, with a strong-layer reinforcement, S increased to (25.2±6.7) and (10.0±1.4) MPa, respectively, at these two porosities. These strengths matched/exceeded the reported strengths of sintered porous hydroxyapatite implants and cancellous bone. Relationships were established between S and the ratio of strong layer thickness/specimen thickness, a/h: S=(17.6 a/h+3.2) MPa. The scaffold contained macropores with a macropore length (mean±sd; n=147) of (183±73) μm, suitable for cell infiltration and tissue ingrowth. Nano-sized hydroxyapatite crystals were observed to form the scaffold matrix of CPC with chitosan. In summary, a layered CPC implant, combining a macroporous CPC with a strong CPC, was developed. Mechanical strength and macroporosity are conflicting requirements. However, the novel functionally graded CPC enabled a relatively high strength and macroporosity to be simultaneously achieved. Such an in situ-hardening nano-apatite may be useful in moderate stress-bearing applications, with macroporosity to enhance tissue ingrowth and implant resorption.

Investigation as to the osteoinductivity of macroporous calcium phosphate cement in goats

For bone formation in critical-sized or poor healing defects, osteoinductive behavior of synthetic bone grafts is crucial. Although the osteoconductive behavior of calcium phosphate (CaP) cement is generally accepted, its osteoinductive potential is less reported. In this study, osteoinduction of porous CaP cement was investigated. Four goats received each six subcutaneous placed prehardened porous CaP cement implants. Implantation time was 3 and 6 months. After explantation, histological evaluation and scoring with a histological grading scale for soft-tissue implants were performed. The histological sections revealed that the implants degraded for more than 50% over time. The implants had lost their macroporous structure from 3 months on. A medium-thick fibrous capsule with a few inflammatory cells surrounded the implants after 3 months. This capsule significantly decreased in thickness after 6 months. Throughout the implant ingrowth of fibrous tissue was seen with scattered foci of inflammatory cells. Cement particles were surrounded by a layer of inflammatory cells. The massive inflammatory response in the interstice was seen after 3 months, which disappeared after 6 months implantation. No bone formation was detected in any of the specimens. The fast degradation and thereby collapsing of the porous structure of our CaP cement implant might have prevented osteoinduction.

Premixed macroporous calcium phosphate cement scaffold

Calcium phosphate cement (CPC) sets in situ to form resorbable hydroxyapatite and is promising for orthopaedic applications. However, it requires on-site powder-liquid mixing during surgery, which prolongs surgical time and raises concerns of inhomogeneous mixing. The objective of this study was to develop a premixed CPC scaffold with macropores suitable for tissue ingrowth. To avoid the on-site powder-liquid mixing, the CPC paste was mixed in advance and did not set in storage; it set only after placement in a physiological solution. Using 30% and 40% mass fractions of mannitol porogen, the premixed CPC scaffold with fibers had flexural strength (mean +/- sd; n = 5) of (3.9 +/- 1.4) MPa and (1.8 +/- 0.8) MPa, respectively. The scaffold porosity reached (68.6 +/- 0.7)% and (74.7 +/- 1.2)%, respectively. Osteoblast cells colonized in the surface macropores of the scaffold and attached to the hydroxyapatite crystals. Cell viability values for the premixed CPC scaffold was not significantly different from that of a conventional non-premixed CPC known to be biocompatible (P > 0.1). In conclusion, using fast-dissolving porogen and slow-dissolving fibers, a premixed macroporous CPC scaffold was developed with strength approaching the reported strengths of sintered porous hydroxyapatite implants and cancellous bone, and non-cytotoxicity similar to a biocompatible non-premixed CPC.

Bone Ingrowth at the Expense of a Novel Macroporous Calcium Phosphate Cement

Bone Ingrowth at the Expense of a Novel Macroporous Calcium Phosphate Cement

Injectable and macroporous calcium phosphate cement scaffold

Calcium phosphate cement (CPC) can be molded and self-hardens in vivo to form resorbable hydroxyapatite with excellent osteoconductivity. The objective of this study was to develop an injectable, macroporous and strong CPC, and to investigate the effects of porogen and absorbable fibers. Water-soluble mannitol was used as porogen and mixed with CPC at mass fractions from 0% to 50%. CPC with 0-40% mannitol was fully extruded under a syringe force of 10 N. The paste with 50% mannitol required a 100-N force which extruded only 66% of the paste. At fiber volume fraction of 0-5%, the paste was completely extruded. However, at 6% and 7.5% fibers, some fibers were left in the syringe after the paste was extruded. The injectable CPC scaffold had a flexural strength (mean+/-sd; n=5) of (3.2+/-1.0) MPa, which approached the reported strengths for sintered porous hydroxyapatite implants and cancellous bone. In summary, the injectability of a ceramic scaffold, a macroporous CPC, was studies for the first time. Processing parameters were tailored to achieve high injectability, macroporosity, and strength. The injectable and strong CPC scaffold may be useful in surgical sites that are not freely accessible by open surgery or when using minimally invasive techniques.

Fast setting calcium phosphate–chitosan scaffold: mechanical properties and biocompatibility

Calcium phosphate cement (CPC) sets in situ to form hydroxyapatite and is highly promising for a wide range of clinical applications. However, its low strength limits its use to only non-stress applications, and its lack of macroporosity hinders cell infiltration, bone ingrowth and implant fixation. The aim of this study was to develop strong and macroporous CPC scaffolds by incorporating chitosan and water-soluble mannitol, and to examine the biocompatibility of the new graft with an osteoblast cell line and an enzymatic assay. Two-way ANOVA identified significant effects on mechanical properties from chitosan reinforcement and powder:liquid ratio ( p<0.001). The flexural strength of CPC–chitosan composite at a powder:liquid ratio of 2 was (13.6±1.2) MPa, which was significantly higher than (3.2±0.6) MPa for CPC control without chitosan (Tukey's at 0.95). At a powder:liquid ratio of 3.5, CPC–chitosan had a strength of (25.3±2.9) MPa, which was significantly higher than (10.4±1.7) MPa for CPC control. The scaffolds possessed total pore volume fractions ranging from 42.0% to 80.0%, and macroporosity up to 65.5%. At total porosities of 52.2–75.2%, the scaffold had strength and elastic modulus values similar to those of sintered porous hydroxyapatite and cancellous bone. Osteoblast mouse cells (MC3T3-E1) were able to adhere, spread and proliferate on CPC–chitosan specimens. The cells, which ranged from about 20 to 50 μm including the cytoplasmic extensions, infiltrated into the 165–271 μm macropores of the scaffold. In summary, substantial reinforcement and macroporosity were imparted to a moldable, fast-setting, biocompatible, and resorbable hydroxyapatite graft. The highly porous scaffold may facilitate bone ingrowth and implant fixation in vivo. In addition, the two to three times increase in strength may help extend the use of CPC to larger repairs in moderately stress-bearing locations.

Preparation of macroporous calcium phosphate cement tissue engineering scaffold

Unlike sintered hydroxyapatite there is evidence to suggest that calcium phosphate cement (CPC) is actively remodelled in vivo and because CPC is formed by a low-temperature process, thermally unstable compounds such as proteins may be incorporated into the matrix of the cement which can then be released after implantation. The efficacy of a macroporous CPC as a bone tissue engineering scaffold has been reported; however, there have been few previous studies on the effect of macroporosity on the mechanical properties of the CPC. This study reports a novel method for the formation of macroporous CPC scaffolds, which has two main advantages over the previously reported manufacturing route: the cement matrix is considerably denser than CPC formed from slurry systems and the scaffold is formed at temperatures below room temperature. A mixture of frozen sodium phosphate solution particles and CPC powder were compacted at 106 MPa and the sodium phosphate was allowed to melt and simultaneously set the cement. The effect of the amount of porogen used during processing on the porosity, pore size distribution and compressive strength of the scaffold was investigated. It was found that macroporous CPC could reliably be fabricated using cement:ice ratios as low as 5:2.

Strong and macroporous calcium phosphate cement: Effects of porosity and fiber reinforcement on mechanical properties.

Because of its excellent osteoconductivity and bone-replacement capability, self-setting calcium phosphate cement (CPC) has been used in a number of clinical procedures. For more rapid resorption and concomitant osseointegration, methods were desired to build macropores into CPC; however, this decreased its mechanical properties. The aims of this study, therefore, were to use fibers to strengthen macroporous CPC and to investigate the effects of the pore volume fraction on its mechanical properties. Water-soluble mannitol crystals were incorporated into CPC paste; the set CPC was then immersed in water to dissolve mannitol, producing macropores. Mannitol/(mannitol + CPC powder) mass fractions of 0, 10, 20, 30, and 40% were used. An aramid fiber volume fraction of 6% was incorporated into the CPC-mannitol specimens, which were set in 3 mm x 4 mm x 25 mm molds and then fractured in three-point flexure to measure the strength, work of fracture, and modulus. The dissolution of mannitol created well-formed macropores, with CPC at 40% mannitol having a total porosity of a 70.8% volume fraction. Increasing the mannitol content significantly decreased the properties of CPC without fibers (analysis of variance; p < 0.001). The strength (mean +/- standard deviation; n = 6) of CPC at 0% mannitol was 15.0 +/- 1.8 MPa; at 40% mannitol, it decreased to 1.4 +/- 0.4 MPa. Fiber reinforcement improved the properties, with the strength increasing threefold at 0% mannitol, sevenfold at 30% mannitol, and nearly fourfold at 40% mannitol. The work of fracture increased by 2 orders of magnitude, but the modulus was not changed as a result of fiber reinforcement. A scanning electron microscopy examination of specimens indicated crack deflection and bridging by fibers, matrix multiple cracking, and frictional pullout of fibers as the reinforcement mechanisms. Macroporous CPCs were substantially strengthened and toughened via fiber reinforcement. This may help extend the use of CPCs with macropores for bony ingrowth to the repair of larger defects in stress-bearing locations.