Porous Calcium Phosphate Cement Research Articles

Design of Porous Calcium Phosphate Cement Composed of Spherical Porous Granules Based on Precipitation of Octacalcium Phosphate

Design of Porous Calcium Phosphate Cement Composed of Spherical Porous Granules Based on Precipitation of Octacalcium Phosphate

Hierarchically porous calcium phosphate scaffold with degradable PLGA microsphere network

Calcium phosphate cement (CPC) is widely used in orthopedics, dentistry and spine surgery because of its excellent biocompatibility, osteoconductivity, arbitrary shaping and self-setting ability. However, slow degradation rate of CPC decreases its bone regeneration efficacy. Herein, poly (lactic co-glycolic acid) microspheres (PLGAm) and wollastonite (WS) were mixed with CPC powder to prepare CPC composite pastes, and then the composite pastes were perfused into poly (lactic co-glycolic acid) network (PLGAnw) to construct composite bone repair materials. The degradation of PLGAnw generated interconnected macropores with the short side of about 468 μm along horizontal axis and the long side of about 785 μm along longitudinal axis, while PLGAm degradation generated isolated or partially connected relative smaller spherical macropores with the size range of 53–106 μm and 106–150 μm, respectively, which was same with PLGAm; in addition, CPC formed micro/nano pores after hydration, thus constructing in-situ hierarchically porous CPC composite scaffolds. Meanwhile, PLGAnw and PLGAm also significantly promoted the degradation of CPC due to the release of acidic by-products. Therefore, the introduction of PLGA network and microspheres not only can evidently accelerate degradation of CPC, but also in-situ form connected macropores and hierarchically porous after their degradation, which are expected to improve bone repair effect of CPC.

Recombinant human bone morphogenetic protein 2/porous calcium phosphate cement/autologous bone and combination of platelet-rich plasma in repairing of large bone defects in rabbits by Masquelet technique

To investigate the optimal mixing ratio of recombinant human bone morphogenetic protein 2 (rhBMP-2) with porous calcium phosphate cement (PCPC) and autologous bone as bone grafting material for the repair of large bone defects using Masquelet technique. The effect of platelet-rich plasma (PRP) on the healing of bone defects was evaluated under the optimal ratio of mixed bone. Fifty-four New Zealand White rabbits were taken to establish a 2 cm long bone defect model of the ulna and treated using the Masquelet technique. Two parts of the experiment were performed in the second phase of the Masquelet technique. First, 36 modeled experimental animals were randomly divided into 4 groups ( n=9) according to the mass ratio of autologous bone and rhBMP-2/PCPC. Group A: autologous bone (100%); group B: 25% autologous bone+75% rhBMP-2/PCPC; group C: 50% autologous bone+50% rhBMP-2/PCPC; group D: 75% autologous bone+25% rhBMP-2/PCPC. The animals were executed at 4, 8, and 12 weeks postoperatively for general observation, imaging observation, histological observation (HE staining), alkaline phosphatase (ALP) activity assay, and biomechanical assay (three-point bending test) were performed to assess the osteogenic ability and to determine the optimal mixing ratio. Then, 18 modeled experimental animals were randomly divided into 2 groups ( n=9). The control group was implanted with the optimal mixture ratio of autologous bone+rhBMP-2/PCPC, and the experimental group was implanted with the optimal mixture ratio of autologous bone+rhBMP-2/PCPC+autologous PRP. The same method was used to observe the above indexes at 4, 8, and 12 weeks postoperatively. The bone healing process from callus formation to the cortical connection at the defected gap could be observed in each group after operation; new bone formation, bridging with the host bone, and bone remodeling to normal bone density were observed on imaging observation; new woven bone, new capillaries, bone marrow cavity, and other structures were observed on histological observation. The ALP activity of each group gradually increased with time ( P<0.05); the ALP activity of group A was significantly higher than that of the other 3 groups at each time point after operation, and of groups C and D than group B ( P<0.05); there was no significant difference between groups C and D ( P>0.05). Biomechanical assay showed that the maximum load in three-point bending test of each group increased gradually with time ( P<0.05), and the maximum loads of groups A and D were significantly higher than that of groups B and C at each time point after operation ( P<0.05), but there was no significant difference between groups A and D ( P>0.05). According to the above tests, the optimal mixing ratio was 75% autogenous bone+25% rhBMP-2/PCPC. The process of new bone formation in the experimental group and the control group was observed by gross observation, imaging examination, and histological observation, and the ability of bone formation in the experimental group was better than that in the control group. The ALP activity and maximum load increased gradually with time in both groups ( P<0.05); the ALP activity and maximum load in the experimental group were significantly higher than those in the control group at each time point after operation ( P<0.05), and the maximum load in the experimental group was also significantly higher than that in group A at 12 weeks after operation ( P<0.05). In the second phase of Masquelet technique, rhBMP-2/PCPC mixed with autologous bone to fill the bone defect can treat large bone defect of rabbit ulna, and it has the best osteogenic ability when the mixing ratio is 75% autologous bone+25% rhBMP-2/PCPC. The combination of PRP can improve the osteogenic ability of rhBMP-2/PCPC and autologous bone mixture.

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.

Fast dissolving glucose porogens for early calcium phosphate cement degradation and bone regeneration

Here, we demonstrate the in vivo efficacy of glucose microparticles (GMPs) to serve as porogens within calcium phosphate cements (CPCs) to obtain a fast-degrading bone substitute material. Composites were fabricated incorporating 20 wt% GMPs at two different GMP size ranges (100–150 μm (GMP-S) and 150–300 μm (GMP-L)), while CPC containing 20 wt% poly(lactic-co-glycolic acid) microparticles (PLGA) and plain CPC served as controls. After 2 and 8 weeks implantation in a rat femoral condyle defect model, specimens were retrieved and analyzed for material degradation and bone formation. Histologically, no adverse tissue response to any of the CPC-formulations was observed. All CPC-porogen formulations showed faster degradation compared to plain CPC control, but only GMP-containing formulations showed higher amounts of new bone formation compared to plain CPC controls. After 8 weeks, only CPC-porogen formulations with GMP-S or PLGA porogens showed higher degradation compared to plain CPC controls. Overall, the inclusion of GMPs into CPCs resulted in a macroporous structure that initially accelerated the generation of new bone. These findings highlight the efficacy of a novel approach that leverages simple porogen properties to generate porous CPCs with distinct degradation and bone regeneration profiles.

Dexamethasone loaded Laponite®/porous calcium phosphate cement for treatment of bone defects

In this research, nanocomposite porous calcium phosphate cements (CPCs) consisting of dexamethasone (DEX) loaded Laponite® nanoplates (LAP) (DEX-LAP) were fabricated and the effects of sodium bicarbonate (NaHCO3) as the foaming agent on the physical, mechanical, and biological characteristics of DEX-LAP loaded CPCs were investigated. Our findings showed that DEX-LAP encapsulation significantly reduced the final setting time (1.5 times) of CPCs and accelerated the total transformation of tricalcium phosphate to hydroxyapatite. Moreover, DEX-LAP nanoplates meaningfully enhanced the compressive strength (4 fold) and compressive modulus (4 fold) of cement. Additionally, incorporation of 6% foaming agent noticeably reduced setting time of cement to less than 10 min due to the enhanced surface area in contact with cement paste. Encapsulation of DEX within the LAP nanoplates and consequently loading of DEX-LAP nanoplates in the CPCs resulted in the significantly reduced burst release of DEX. Finally, the results also illustrated that the presence of interconnected micropores and DEX-LAP nanoplates in the CPC improved MG63 cell proliferation. These findings suggest that the porous DEX-LAP/CPC can potentially be developed as bone grafts for bone defects.

Novel Strategy to Accelerate Bone Regeneration of Calcium Phosphate Cement by Incorporating 3D Plotted Poly(lactic-co-glycolic acid) Network and Bioactive Wollastonite.

Inefficient bone regeneration of self-hardening calcium phosphate cement (CPC) increases the demand for interconnected macropores and osteogenesis-stimulated substances. It remains a challenge to fabricate porous CPC with interconnected macropores while maintaining its advantages, such as plasticity. Herein, pastes containing CPC and wollastonite (WS) are infiltrated into a 3D plotted poly(lactic-co-glycolic acid) (PLGA) network to fabricate plastic CPC-based composite cement (PLGA/WS/CPC). The PLGA/WS/CPC recovers the plasticity of CPC after being heated above the glass transition temperature of PLGA. The presence of the 3D PLGA network significantly increases the flexibility of CPC in prophase and generates 3D interconnected macropores in situ upon its degradation. The addition of WS is helpful to improve the attachment, proliferation, and osteogenic differentiation of mouse bone marrow stromal cells in vitro. The in vivo experimental results indicate that PLGA/WS/CPC promotes rapid angiogenesis and bone formation. Therefore, the plastic CPC-based composite cement with a 3D PLGA network and wollastonite shows an obviously improved efficiency for repairing bone defects and is expected to facilitate the wider application of CPC in the clinic.

Experiment of porous calcium phosphate/bone matrix gelatin composite cement for repairing lumbar vertebral bone defect in rabbit

To investigate the effect of a porous calcium phosphate/bone matrix gelatin (BMG) composite cement (hereinafter referred to as the "porous composite cement") for repairing lumbar vertebral bone defect in a rabbit model. BMG was extracted from adult New Zealand rabbits according to the Urist's method. Poly (lactic-co-glycolic) acid (PLGA) microsphere was prepared by W/O/W double emulsion method. The porous composite cement was developed by using calcium phosphate cement (CPC) composited with BMG and PLGA microsphere. The physicochemical characterizations of the porous composite cement were assessed by anti-washout property, porosity, and biomechanical experiment, also compared with the CPC. Thirty 2-month-old New Zealand rabbits were used to construct vertebral bone defect at L 3 in size of 4 mm×3 mm×3 mm. Then, the bone defect was repaired with porous composite cement (experimental group, n=15) or CPC (control group, n=15). At 4, 8, and 12 weeks after implantation, each bone specimen was assessed by X-ray films for bone fusion, micro-CT for bone mineral density (BMD), bone volume fraction (BVF), trabecular thickness (Tb. Th.), trabecular number (Tb.N.), and trabecular spacing (Tb. Sp.), and histological section with toluidine blue staining for new-born bone formation. The study demonstrated well anti-washout property in 2 groups. The porous composite cement has 55.06%±1.18% of porosity and (51.63±6.73) MPa of compressive strength. The CPC has 49.38%±1.75% of porosity and (63.34±3.27) MPa of compressive strength. There were significant differences in porosity and compressive strength between different cements ( t=4.254, P=0.006; t=2.476, P=0.034). X-ray films revealed that the zone between the cement and host bone gradually blurred with the time extending. At 12 weeks after implantation, the zone was disappeared in the experimental group, but clear in the control group. There were significant differences in BMD, BVF, Tb. Th., Tb. N., and Tb. Sp. between 2 groups at each time point ( P<0.05). Histological observation revealed that there was new-born bone in the cement with the time extending in 2 groups. Among them, bony connection was observed between the new-born bone and the host in the experimental group, which was prior to the control group. The porous composite cement has dual bioactivity of osteoinductivity and osteoconductivity, which are effective to promote bone defect healing and reconstruction.

3D-printing of porous calcium phosphate cements for bone tissue engineering

3D-printing of porous calcium phosphate cements for bone tissue engineering

Mechanical Activation Reinforced Porous Calcium Phosphate Cement

Calcium phosphate cement (CPC) powder was activated by ball milling to improve the mechanical properties of porous CPC scaffolds. The mechanical activation mechanism was investigated by specific surface analyses, X-ray diffraction (XRD) and scanning electron microscopy (SEM). After ball milling, the average particle sizes of BCPC powder decreased while the specific surface area, apparent density, bulk density, and compact density increased when compared with non-activated CPC powder. The porosity and compressive strength of porous CPC scaffolds prepared from ball-milled powders (BCPC-S) were (77.98 ± 0.58)% and (4.11 ± 0.46) MPa, both significantly higher than those non-activated CPC powders (CPC-S), whose porosity and compressive strength were (64.23  2.32)% and (1.99 ± 0.43) MPa, respectively. SEM revealed that there were two types of pores in the BCPC-S: one ranged a few microns in size and the other ranged several hundred microns. XRD indicated that grain sizes and crystallinities of dicalcium phosphate dehydrate (DCPD), α-tricalcium phosphate (α-TCP), calcium carbonate (CaCO3) and hydroxyapatite (HA) in BCPC powder decreased, due to the mechanical activation compared to 第 4期 黄 萍, 等: 机械活化增强多孔磷酸钙骨水泥支架的研究 433 those of the non-activated CPC powders. In addition, the mechanical activation resulted in the conversion of DCPD to dicalcium phosphate anhydrous (DCPA), which promoted the hydration of CPC and the precipitation of HA, and improved the compressive strength of BCPC-S finally. This study provided a potential approach to improve the mechanical properties of porous CaP based scaffold to meet the clinic requirement.

Effects of adding resorbable phosphate glass fibres and PLA to calcium phosphate bone cements.

Calcium phosphate cements (CPCs), due to their biocompatibility and degradation properties, are being widely investigated as a replacement to more commonly used polymethylmethacrylate (PMMA) for vertebroplasty. CPCs have shown the potential to be replaced by host bone tissue during the healing/remodelling process. However, brittleness and comparatively low strength restrict the use of CPC in load-bearing applications. Although porous CPC can integrate with bone over time, slow degradation profiles and poor interconnectivity between pores restricts osseointegration to the top layer of CPC only. Polylactic acid (PLA) and phosphate glass fibres (PGFs) were incorporated in a CPC matrix to overcome the problem of inherent brittleness and limited osseointegration. Incorporation of PLA and PGFs within CPC was successful in achieving a much less brittle CPC matrix without affecting the mechanical properties of CPC. The area under the stress-strain curve showed that the total energy to failure of the CPC hybrid was significantly greater than that of the CPC control. The methodology adopted here to add PLA within the CPC matrix may also allow for incorporation of PLA cross-linked biochemicals. Micrographic studies revealed that it was possible to confer control over pore size, shape and interconnectivity without negatively affecting the mechanical properties of the cement. This tailorable porosity could potentially lead to better osseointegration within CPC.

Monitoring the biological effect of BMP-2 release on bone healing by PET/CT

Bone morphogenetic protein-2 (BMP-2) releasing scaffolds are routinely used in bone tissue engineering to accelerate regeneration. Thus, assessing growth factor activity, and tissue regeneration around a biomaterial delivery system is extremely important to optimize the stages of bone healing. Such assessment commonly relies on histology. A non-invasive method would allow longitudinal monitoring of the regeneration process. The present study sought to explore the feasibility of Positron Emission Tomography (PET) as a non-invasive method to monitor the osteogenic potential of a BMP-2 releasing calcium phosphate cement (CPC) bone substitute. In a calvarial defect model in rats, 18F-Fluoride PET was used to quantify 18F-Fluoride uptake around the BMP-2/CPC material. As controls, non-loaded porous CPC, dense CPC and autograft samples were used. PET was performed every two weeks, during a period of 8weeks post-implantation. In parallel, computed tomography (CT) imaging was performed for anatomical reference. Four specimens were evaluated per group per time point. The highest 18F-Fluoride signal intensity was measured in the BMP-2 group (approximately 5 times higher than the baseline value), followed by the controls in the order, from high to low, porous, dense and autograft. The same trend was maintained during the entire experimental period. Histology and measurements of the bone volume percentage confirmed the significantly higher new bone formation in the BMP-2 releasing group (about 25%), compared to the controls. Moreover, 18F-Fluoride PET uptake showed a statistically significant positive correlation (r=0.6038, P=0.0171) with the newly formed bone volume. In conclusion, our results support the use of a PET/CT method for the longitudinal monitoring of BMP-2 releasing scaffolds, in vivo.

The releasing behavior and in vitro osteoinductive evaluations of dexamethasone-loaded porous calcium phosphate cements

In the present study, the effect of adding different concentrations of dexamethasone (DEX) on setting time, apatite formation and mechanical properties of micro/macroporous calcium phosphate cements (CPCs) was studied. The liberation of loaded DEX was evaluated over 0–840 h in simulated body fluid (SBF). Differentiation of mesnchymal stem cells on DEX-free and loaded CPCs were also compared by expression of bone marker proteins. The results showed that setting time decreased by adding DEX to CPCs whereas mechanical strength and conversion rate of cement reactants into apatite phase. A sustained release of DEX was observed from the loaded CPCs in which the rate of DEX release increased by introducing macroporosity and higher portion of loaded DEX. Osteogenic differentiation of MSCs on DEX-loaded CPCs was confirmed by elevated gene expressions of bone markers such as alkaline phosphatase, osteopontin and osteocalcin. Overall, the results reveal that DEX-loaded CPCs may be potentially osteoinductive material with enhanced biological functions but more in vivo evaluations are required to prove this suggestion.

Enhanced Dentin-Like Mineralized Tissue Formation by AdShh-Transfected Human Dental Pulp Cells and Porous Calcium Phosphate Cement

The aim of the present study was to investigate the effect of Sonic hedgehog (Shh) on human dental pulp cells (hDPCs) and the potential of complexes with Shh gene modified hDPCs and porous calcium phosphate cement (CPC) for mineralized tissue formation. hDPCs were cultured and transfected with adenoviral mediated human Shh gene (AdShh). Overexpression of Shh and cell proliferation was tested by real-time PCR analysis, western blotting analysis, and MTT analysis, respectively. The odontoblastic differentiation was assessed by alkaline phosphatase (ALP) activity and real-time PCR analysis on markers of Patched-1 (Ptc-1), Smoothened (Smo), Gli 1, Gli 2, Gli 3, osteocalcin (OCN), dentin matrix protein-1 (DMP-1), and dentin sialophosphoprotein (DSPP). Finally, AdShh-transfected hDPCs were combined with porous CPC and placed subcutaneously in nude mice for 8 and 12 weeks, while AdEGFP-transfected and untransfected hDPCs were treated as control groups. Results indicated that Shh could promote proliferation and odontoblastic differentiation of hDPCs, while Shh/Gli 1 signaling pathway played a key role in this process. Importantly, more mineralized tissue formation was observed in combination with AdShh transfected hDPCs and porous CPC, moreover, the mineralized tissue exhibited dentin-like features such as structures similar to dentin-pulp complex and the positive staining for DSPP protein similar to the tooth tissue. These results suggested that the constructs with AdShh-transfected hDPCs and porous CPC might be a better alternative for dental tissue regeneration.

Bi-layered calcium phosphate cement-based composite scaffold mimicking natural bone structure

In this study, a core/shell bi-layered calcium phosphate cement (CPC)-based composite scaffold with adjustable compressive strength, which mimicked the structure of natural cortical/cancellous bone, was fabricated. The dense tubular CPC shell was prepared by isostatic pressing CPC powder with a specially designed mould. A porous CPC core with unidirectional lamellar pore structure was fabricated inside the cavity of dense tubular CPC shell by unidirectional freeze casting, followed by infiltration of poly(lactic-co-glycolic acid) and immobilization of collagen. The compressive strength of bi-layered CPC-based composite scaffold can be controlled by varying thickness ratio of dense layer to porous layer. Compared to the scaffold without dense shell, the pore interconnection of bi-layered scaffold was not obviously compromised because of its high unidirectional interconnectivity but poor three dimensional interconnectivity. The in vitro results showed that the rat bone marrow stromal cells attached and proliferated well on the bi-layered CPC-based composite scaffold. This novel bi-layered CPC-based composite scaffold is promising for bone repair.

Engineering of Bone Using Porous Calcium Phosphate Cement and Bone Marrow Stromal Cells for Maxillary Sinus Augmentation with Simultaneous Implant Placement in Goats

The aim of this study was to explore the effects of maxillary sinus floor elevation and simultaneous dental implantation with a tissue-engineered bone complex of calcium phosphate cement (CPC) scaffolds combined with bone marrow stromal cells (BMSCs). A large animal goat model is used with the tissue engineering method. Eighteen bilateral maxillary sinus of nine goats were randomly allocated into three groups; the CPC/BMSC complex (n=6) was used to elevate maxillary sinus floor with a simultaneous implant placement; the effects were compared with those treated with CPC alone (n=6) or autogenous bone (n=6). After a healing period of 3 months, sequential triad-color fluorescence labeling, micro-CT, as well as histological and histomorphometric analyses indicated that the tissue-engineered BMSC/CPC complex could promote earlier bone formation and mineralization, and maximally maintain the volume and height of the augmented maxillary sinus. By comparison, CPC-alone or autogenous bone achieved less bone formation and later mineralization. Besides, the average bone-implant contact value reflecting the osseointegration was 35.63%±9.42% in the BMSCs/CPC group, significantly higher than 22.47%±4.28% in the CPC-alone group or 28.26%±8.03% in the autogenous bone group. In conclusion, CPC serves as a potential substrate for BMSCs for the maxillary sinus floor augmentation and simultaneous implantation. The tissue-engineered bone might enhance the stability of implants and thus be of great significance to achieve improved quality to restore the oral function in clinic.

Three Different Strategies to Obtain Porous Calcium Phosphate Cements: Comparison of Performance in a Rat Skull Bone Augmentation Model

Preprosthetic surgery has become a routine procedure to obtain sufficient bone quantity and quality for dental implant installation in patients with an initial inadequate bone volume. Although autologous bone onlay or inlay grafting is still the preferred bone augmentation technique, a broad range of synthetic bone substitutes have been developed, for example, calcium phosphate cement (CPC). The introduction of porosity within CPC can be used to increase CPC degradation and bone ingrowth. Therefore, three different strategies to obtain porous CPCs were evaluated in this preclinical study. Instantaneously porous CPC (CPC-IP) was compared with delayed porous CPC in vitro and in vivo. CPC-IP was obtained by the creation of CO₂ bubbles during setting, whereas delayed porous CPC was obtained after the degradation of incorporated poly(lactic-co-glycolic acid) (PLGA) microspheres. As an additional aspect, delayed porous CPC was created by the incorporation of either hollow or dense degradable PLGA microspheres (CPC-hPLGA and CPC-dPLGA). All CPC compositions showed appropriate clinical handling properties and an interconnected porous structure with a final porosity above 70% (v/v). In vitro degradation studies showed the gradual formation of pores and further CPC-matrix dissolution for CPCs containing PLGA microspheres (dPLGA microspheres > hPLGA microspheres). For in vivo evaluation of the CPCs, an augmentation model was used, allowing a CPC injection into a rigidly immobilized Teflon ring on the rat skull. Histological evaluation after 12 weeks of implantation showed bone formation using all three CPCs. Bone apposition reached volumetric amounts of up to 10% of the augmentation area and a maximum augmentation height of ∼1 mm. CPC-IP showed significantly more bone formation and resulted in a superior bone apposition height compared with both CPCs containing PLGA microspheres. No differences in biological performance were observed between the CPCs containing hPLGA and those containing dPLGA microspheres. Further research is necessary to enhance the bone appositional speed and amount of CPCs for bone augmentation procedures before them being used in a potential clinical setting.

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.

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.

LvBMP-2 gene-modified BMSCs combined with calcium phosphate cement scaffolds for the repair of calvarial defects in rats

The study aims to evaluate the effect of bone marrow stromal cells (BMSCs) expressing bone morphogenic protein-2 (BMP-2) mediated by lentiviral (Lv) gene transduction combined with calcium phosphate cement (CPC) scaffolds for the repair of critical size calvarial defects in rats. BMSCs derived from Fisher 344 rats were transduced with LvBMP-2 or lentivirus encoding enhanced green fluorescent protein (LvEGFP) in vitro. Obvious osteogenic differentiation of BMSCs in the LvBMP-2 group was demonstrated by alkaline phosphatase staining and alizarin red staining. Enzyme-linked immunosorbent assay results show that LvBMP-2 gene expression in vitro can last for at least 8weeks. Gene-transduced or untransduced BMSCs were seeded onto CPC scaffolds to repair rat calvarial defects with a diameter of 5mm. Scanning electron microscope analysis indicated that porous CPC scaffolds facilitated initial adhesion and spreading of BMSCs onto its surface. Calvarial defects were successfully repaired with LvBMP-2-transduced BMSCs/CPC constructs 8weeks postoperatively. The percentage of new bone formation in the LvBMP-2 group was significantly higher than in other control groups. Lentiviral mediated BMP-2 gene therapy together with CPC scaffolds can be used successfully in calvarial repair and bone regeneration.