Degradation Of Calcium Phosphate Cement Research Articles

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

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

Injectable and high-strength PLGA/CPC loaded ALN/MgO bone cement for bone regeneration by facilitating osteogenesis and inhibiting osteoclastogenesis in osteoporotic bone defects

Osteoporosis (OP) can result in slower bone regeneration than the normal condition due to the imbalance between osteogenesis and osteoclastogenesis, making osteoporotic bone defects healing a significant clinical challenge. Calcium phosphate cement (CPC) is a promising bone substitute material due to its good osteoinductive activity, however, the drawbacks such as fragility, slow degradation rate and incapability to control bone loss restrict its application in osteoporotic bone defects treatment. Currently, we developed the PLGA electrospun nanofiber sheets to carry alendronate (ALN) and magnesium oxide nanoparticle (nMgO) into CPC, therefore, to obtain a high-strength bone cement (C/AM-PL/C). The C/AM-PL/C bone cement had high mechanical strength, anti-washout ability, good injection performance and drug sustained release capacity. More importantly, the C/AM-PL/C cement promoted the osteogenic differentiation of bone marrow mesenchymal stem cells and neovascularization via the release of Mg2+ (from nMgO) and Ca2+ (during the degradation of CPC), and inhibited osteoclastogenesis via the release of ALN in vitro. Moreover, the injection of C/AM-PL/C cement significantly improved bone healing in an OP model with femur condyle defects in vivo. Altogether, the injectable C/AM-PL/C cement could facilitate osteoporotic bone regeneration, demonstrating its capacity as a promising candidate for treatment of osteoporotic bone defects.

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.

Early-stage macroporosity enhancement in calcium phosphate cements by inclusion of poly(N-vinylpyrrolidone) particles as a porogen

The incorporation of poly(DL-lactic-co-glycolic acid) (PLGA) particles into calcium phosphate cements (CPCs) is an effective strategy to enhance CPC macroporosity and degradation. However, bone regeneration is hindered until hydrolytic PLGA degradation starts a few weeks after implantation. Additionally, CPC and CPC/PLGA injectability and cohesion are suboptimal. In the current study, poly(N-vinylpyrrolidone) (PVP), a water-soluble polymer, was incorporated as a porogen in CPC and CPC/PLGA composites to enhance handling properties and early-stage macroporosity formation. Further, the effect of PVP molecular weight (Mw) and particle size was studied. The results showed that PVP incorporation increased both injectability and cohesion of the CPC pastes, especially with addition of high Mw PVP. Moreover, the in vitro degradation studies revealed that incorporation of PVP induced an initial mass loss during the first week of incubation. In combination with PLGA, small PVP particles induced a higher mass loss at an early stage than large PVP particles, but this effect was no longer apparent after 4 weeks of incubation. In contrast, the incorporation of low Mw PVP had a stronger effect on in vitro degradation in the long term compared to high Mw. Finally, the presence of PLGA porogens appeared to be necessary for adequate CPC degradation.

Multimodal porogen platforms for calcium phosphate cement degradation.

Calcium phosphate cements (CPCs) represent excellent bone substitute materials due to their biocompatibility and injectability. However, their poor degradability and lack of macroporosity limits bone regeneration. The addition of poly(d,l‐lactic‐co‐glycolic acid) (PLGA) particles improves macroporosity and therefore late stage material degradation. CPC degradation and hence, bone formation at an early stage remains challenging, due to the delayed onset of PLGA degradation (i.e., after 2–3 weeks). Consequently, we here explored multimodal porogen platforms based on sucrose porogens (for early pore formation) and PLGA porogens (for late pore formation) to enhance CPC degradation and analyzed mechanical properties, dynamic in vitro degradation and in vivo performance in a rat femoral bone defect model. Porogen addition to CPC showed to decrease compressive strength of all CPC formulations; transition of the crystal phase upon in vitro incubation increased compressive strength. Although dynamic in vitro degradation showed rapid sucrose dissolution within 1 week, no additional effects on CPC degradation or bone formation were observed upon in vivo implantation. © 2019 The Authors. journal Of Biomedical Materials Research Part A Published By Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 1713–1722, 2019.

Effect of Low-Frequency Pulsed Ultrasound on Drug Delivery, Antibacterial Efficacy, and Bone Cement Degradation in Vancomycin-Loaded Calcium Phosphate Cement.

BackgroundCalcium phosphate cement (CPC) has been applied as a biodegradable antibiotic carrier in osteomyelitis. However, the drug delivery, antibacterial efficacy, and degradation rate of CPC are insufficient and require further improvement in clinical application.Material/MethodsVancomycin-loaded CPC columns were prepared, and eluted in simulated body fluid. The drug delivery was assessed in the ultrasound group and control group by fluorescence polarization immunoassay. The antibacterial efficacy of vancomycin in the ultrasound group and control groups was investigated by standard plate count method. Low-frequency pulsed ultrasound (46.5 kHz, 900 mW/cm2) was used to produce a sinusoidal wave in the ultrasound groups. The percentage of residual weight was evaluated to assess the degradation of CPC.ResultsThe concentration and cumulatively released percentage of vancomycin in the ultrasound group were higher than that in the control group at each time point (p<0.05). The duration of vancomycin concentration over the level of minimum inhibitory concentration was significantly prolonged in the ultrasound group (p<0.05). Antibacterial efficacy of vancomycin in the ultrasound group was significantly greater than that in the control group with same concentration of vancomycin (p<0.05). The percentage of residual weight in the ultrasound group was significantly less than that in the control group (p<0.05).ConclusionsLow-frequency pulsed ultrasound can enhance vancomycin release, prolong the duration of vancomycin concentration at high levels, and accelerate the degradation rate of vancomycin-loaded CPC.

Multimodal pore formation in calcium phosphate cements.

Calcium phosphate cements (CPCs) are commonly used as bone substitute materials. However, their slow degradation rate and lack of macroporosity hinders new bone formation. Poly(dl-lactic-co-glycolic acid) (PLGA) incorporation is of great interest as, upon degradation, produces acidic by-products that enhance CPC degradation. Yet, new bone formation is delayed until PLGA degradation occurs a few weeks after implantation. Therefore, the aim of this study was to accelerate the early stage pore formation within CPCs in vitro. With that purpose, we incorporated the water-soluble porogen sucrose at different weight percentages (10 or 20 wt %) to CPC and CPC/PLGA composites. The results revealed that incorporation of sucrose porogens increased mass loss within the first week of in vitro degradation in groups containing sucrose compared to control groups. After week 1, a further mass loss was observed related to PLGA and CPC degradation. Macroporosity analysis confirmed that macroporosity formation is influenced by the dissolution of sucrose at an early stage and by the degradation of PLGA and CPC at a later stage. We concluded that the combination of sucrose and PLGA porogens in CPC is a promising approach to promote early stage bone tissue ingrowth and complete replacement of CPC through multimodal pore formation. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 500-509, 2018.

Salmon calcitonin‐loaded PLGA microspheres/calcium phosphate cement composites for osteoblast proliferation

ABSTRACTTo improve the efficacy of salmon calcitonin (SCT) on disuse osteoporosis‐induced bone fracture, the sustained‐release vehicles delivered to bone fractures should be developed. In this study, SCT‐loaded poly(d,l‐lactic‐co‐glycolic acid) microspheres (SCT‐PLGA MS) with 20 kDa‐COOH and 40 kDa‐COOH (SCT‐PLGA‐20 COOH MS and SCT‐PLGA‐40 COOH MS) are incorporated into calcium phosphate cement (CPC) at various mass ratios. The cumulative release and mechanical properties of SCT‐PLGA MS/CPC composites decrease as PLGA MS/CPC mass ratio increase. Scanning electron microscopy images and the mass loss of composites indicate the MS from 20% SCT‐PLGA MS/CPC composites have mostly degraded after 8 weeks. In addition, SCT released from the 20% SCT‐PLGA‐20 MS/CPC composites significantly facilitate proliferation and differentiation of MC3T3‐E1 cells. In conclusion, 20% SCT‐PLGA‐20 COOH MS/CPC composites exhibit the sustained drug release, proper CPC degradation, good mechanical properties, and preferable osteoblast proliferation, which would be beneficial to their in vivo applications. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017, 134, 45486.

Perfluorocarbon/Gold Loading for Noninvasive in Vivo Assessment of Bone Fillers Using 19F Magnetic Resonance Imaging and Computed Tomography.

Calcium phosphate cement (CPC) is used in bone repair because of its biocompatibility. However, high similarity between CPC and the natural osseous phase results in poor image contrast in most of the available in vivo imaging techniques such as computed tomography (CT) and magnetic resonance imaging (MRI). For accurate identification and localization during and after implantation in vivo, a composition with enhanced image contrast is needed. In this study, we labeled CPC with perfluoro-15-crown-5-ether-loaded (PFCE) poly(latic-co-glycolic acid) nanoparticles (hydrodynamic radius 100 nm) and gold nanoparticles (diameter 40 nm), as 19F MRI and CT contrast agents, respectively. The resulting CPC/PFCE/gold composite is implanted in a rat model for in vivo longitudinal imaging. Our findings show that the incorporation of the two types of different nanoparticles did result in adequate handling properties of the cement. Qualitative and quantitative long-term assessment of CPC/PFCE/gold degradation was achieved in vivo and correlated to the new bone formation. Finally, no adverse biological effects on the bone tissue are observed via histology. In conclusion, an easy and efficient strategy for following CPC implantation and degradation in vivo is developed. As all materials used are biocompatible, this CPC/PFCE/gold composite is clinically applicable.

Preparation and characterization of calcium phosphate bone cement with rapidly-generated tubular macroporous structure by incorporation of polysaccharide-based microstrips

Calcium phosphate cements (CPCs) have been extensively used as bone graft substitutes for the repair of bone defect due to its biocompatibility, osteoconductivity and in-situ setting capability. They poorly degrade thus limiting their use in tissue engineering application. A possible strategy to improve the speed of CPC degradation is to add porogen to CPC to create macropores that can enhance cement resorption and can consequently be replaced by new bone. The as-generated macropores are generally not connected because of spherical shape of the porogens which can limit the extent of newly formed bone. The aim of this study was to fabricate CPCs having tubular macroporous structure by incorporating fast-dissolving maltodextrin microstrips (MDMS) and explore their properties such as setting time, mechanical property, microstructure and degradability of the cements. The results showed that after immersing MDMS-embedded composites in simulated body fluid under physiological condition for 1d MDMS rapidly disintegrated (more than 70%), generating tubular macropores in CPCs. The disintegration of MDMS completed in 1 week. CPCs containing MDMS lower than 30% by weight had the same final setting time as those without MDMS. The average values of compressive strength of the CPC composites decreased with the disintegration of MDMS. % Porosity and pore interconnectivity increased with increasing MDMS content. In addition, MDMS-embedded CPCs were cell friendly with excellent cell adhesion, indicating a possible candidate as bone graft substitutes.

Long-term evaluation of the degradation behavior of three apatite-forming calcium phosphate cements.

Calcium phosphate cements (CPCs) are injectable bone substitutes with a long clinical history because of their biocompatibility and osteoconductivity. Nevertheless, their cohesion upon injection into perfused bone defects as well as their long-term degradation behavior remain major clinical challenges. Therefore, the long-term degradation behavior of two types of α-tricalcium phosphate-based, apatite-forming CPCs was compared to a commercially available apatite-forming cement, that is HydroSet™ . Carboxyl methylcellulose (CMC) was used as cohesion promotor to improve handling properties of the two experimental cements, whereas poly (d, l-lactic-co-glycolic) acid (PLGA) microparticles were added to introduce macroporosity and stimulate CPC degradation. All three CPCs were injected into defects drilled into rabbit femoral condyles and explanted after 4, 12, or 26 weeks, after which the bone response was assessed both qualitatively and quantitatively. CPCs without PLGA microparticles degraded only at the periphery of the implants, while the residual CPC volume was close to 90%. On the contrary, bone ingrowth was observed not only at the periphery of the CPC, but also throughout the center of the implants after 26 weeks of implantation for the PLGA-containing CPCs with a residual CPC volume of approximately 55%. In conclusion, it was shown that CPC containing CMC and PLGA was able to induce partial degradation of apatite-forming CPCs and concomitant replacement by bone tissue.

Effect of ultrafine poly(ε-caprolactone) fibers on calcium phosphate cement: in vitro degradation and in vivo regeneration.

We incorporated ultrafine polymer fibers into calcium phosphate cement (CPC) to improve the resorption rate of CPC with fiber degradation. Different weight percentages of electrospun poly(ε-caprolactone) fibers (0%, 3%, and 7%, named as ultrafine fiber-incorporated CPC0 [UFICPC0], UFICPC3, and UFICPC7) were included into preset CPC specimens for in vitro immersion in lipase phosphate-buffered solution and long-term in vivo implantation in the femoral condyle of rabbits. The effect of the ultrafine poly(ε-caprolactone) fibers with a diameter ranging from nanometer to micrometer on CPC degradation was evaluated by measuring the pH of the medium, mass loss, porosity, and physiochemical properties. For the in vivo evaluation, histomorphometrical analysis as well as three-dimensional (3D) reconstruction was applied to assess the osteogenic properties of the CPC composite. After in vitro immersion and in vivo implantation, the total porosity and macroporosity as well as the bone formation and ingrowth increased significantly during time in the fiber-incorporated CPC specimens. After 24 weeks of implantation, the degraded space was occupied by newly formed bone, and the UFICPC3 and UFICPC7 composites showed ~3.5 times higher fraction of bone volume than that of the pristine CPC (UFICPC0). In vitro and in vivo results proved that the introduction of ultrafine degradable fibers within a CPC matrix can be used to improve macroporosity efficiently and enhance CPC degradation and bone ingrowth largely.

Electrospun Nanofibrous P(DLLA-CL) Balloons as Calcium Phosphate Cement Filled Containers for Bone Repair: in Vitro and in Vivo Studies.

The spinal surgeon community has expressed significant interest in applying calcium phosphate cement (CPC) for the treatment of vertebral compression fractures (VCFs) and minimizing its disadvantages, such as its water-induced collapsibility and poor mechanical properties, limiting its clinical use. In this work, novel biodegradable electrospun nanofibrous poly(d,l-lactic acid-ϵ-caprolactone) balloons (ENPBs) were prepared, and the separation, pressure, degradation, and new bone formation behaviors of the ENPBs when used as CPC-filled containers in vitro and in vivo were systematically analyzed and compared. CPC could be separated from surrounding bone tissues by ENPBs in vitro and in vivo. ENPB-CPCs (ENPBs serving as CPC-filled containers) exerted pressure on the surrounding bone microenvironment, which was enough to crush trabecular bone. Compared with the CPC implantation, ENPB-CPCs delayed the degradation of CPC (i.e., its water-induced collapsilibity). Finally, possible mechanisms behind the in vivo effects caused by ENPB-CPCs implanted into rabbit thighbones and pig vertebrae were proposed. This work suggests that ENPBs can be potentially applied as CPC-filled containers in vivo and provides an experimental basis for the clinical application of ENPBs for the treatment of VCFs. In addition, this work will be of benefit to the development of polymer-based medical implants in the future.

A theranostic agent to enhance osteogenic and magnetic resonance imaging properties of calcium phosphate cements

With biomimetic biomaterials, like calcium phosphate cements (CPCs), non-invasive assessment of tissue regeneration is challenging. This study describes a theranostic agent (TA) to simultaneously enhance both imaging and osteogenic properties of such a bone substitute material. For this purpose, mesoporous silica beads were produced containing an iron oxide core to enhance bone magnetic resonance (MR) contrast. The same beads were functionalized with silane linkers to immobilize the osteoinductive protein BMP-2, and finally received a calcium phosphate coating, before being embedded in the CPC. Both in vitro and in vivo tests were performed. In vitro testing showed that the TA beads did not interfere with essential material properties like cement setting. Furthermore, bioactive BMP-2 could be efficiently released from the carrier-beads. In vivo testing in a femoral condyle defect rat model showed long-term MR contrast enhancement, as well as improved osteogenic capacity. Moreover, the TA was released during CPC degradation and was not incorporated into the newly formed bone. In conclusion, the described TA was shown to be suitable for longitudinal material degradation and bone healing studies.

Zero Echo Time Magnetic Resonance Imaging of Contrast-Agent-Enhanced Calcium Phosphate Bone Defect Fillers

Calcium phosphate cements (CPCs) are widely used bone substitutes. However, CPCs have similar radiopacity as natural bone, rendering them difficult to be differentiated in classical X-ray and computed tomography imaging. As conventional magnetic resonance imaging (MRI) of bone is cumbersome, due to low water content and very short T(2) relaxation time, ultra-short echo time (UTE) and zero echo time (ZTE) MRI have been explored for bone visualization. This study examined the possibility to differentiate bone and CPC by MRI. T(1) and T(2)* values determined with UTE MRI showed little difference between bone and CPC; hence, these materials were difficult to separate based on T(1) or T(2) alone. Incorporation of ultra-small particles of iron oxide and gadopentetatedimeglumine (Gd-DTPA; 1 weight percentage [wt%] and 5 wt% respectively) into CPC resulted in visualization of CPC with decreased intensity on ZTE images in in vitro and ex vivo experiments. However, these additions had unfavorable effects on the solidification time and/or mechanical properties of the CPC, with the exception of 1% Gd-DTPA alone. Therefore, we tested this material in an in vivo experiment. The contrast of CPC was enhanced at an early stage postimplantation, and was significantly reduced in the 8 weeks thereafter. This indicates that ZTE imaging with Gd-DTPA as a contrast agent could be a valid radiation-free method to visualize CPC degradation and bone regeneration in preclinical experiments.

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.

Wollastonite nanofiber&amp;ndash;doped self-setting calcium phosphate bioactive cement for bone tissue regeneration

The purpose of this study was to synthesize a self-setting bioactive cement by incorporation of wollastonite nanofibers (WNFs) into calcium phosphate cement (CPC). The composition, morphology, setting time, compressive strength, hydrophilicity, and degradation of WNF-doped CPC (wnf-CPC) were investigated. Scanning electron microscopy, Fourier transform infrared spectroscopy, X-ray diffraction, and inductively coupled plasma atomic emission spectroscopy were utilized. Additionally, methyl-thiazolyl-tetrazolium bromide assay, scanning electron microscopy, inductively coupled plasma atomic emission spectroscopy, and histological evaluation were used to study the cell and tissue responses to wnf-CPC, both in vitro and in vivo. The results confirmed that the addition of WNFs into CPC had no obvious effect on the setting time or the compressive strength of wnf-CPC, provided the WNF amount was not more than 10 wt%. However, the hydrophilicity and degradability of wnf-CPC were significantly improved by the addition of WNFs – this was because of the change of microstructure caused by the WNFs. The preferred dissolution of WNFs caused the formation of microporosity in wnf-CPC when soaked in tris hydrochloride solution. The microporosity enlarged the surface area of the wnf-CPC and so promoted degradation of the wnf-CPC when in contact with liquid. In addition, MG-63 cell attachment and proliferation on the wnf-CPC were superior to that on the CPC, indicating that incorporation of WNFs into CPC improved the biological properties for wnf-CPC. Following the implantation of wnf-CPC into bone defects of rabbits, histological evaluation showed that wnf-CPC enhanced the efficiency of new bone formation in comparison with CPC, indicating excellent biocompatibility and osteogenesis of wnf-CPC. In conclusion, wnf-CPC exhibited promising prospects in bone regeneration.

Bone response to fast-degrading, injectable calcium phosphate cements containing PLGA microparticles

Apatitic calcium phosphate cements (CPC) are frequently used to fill bone defects due to their favourable clinical handling and excellent bone response, but their lack of degradability inhibits complete bone regeneration. In order to render these injectable CaP cements biodegradable, hollow microspheres made of poly ( d, l-lactic-co-glycolic) acid (PLGA) have been previously used as porogen since these microspheres were shown to be able to induce macroporosity upon degradation as well as to accelerate CPC degradation by release of acid degradation products. Recently, the capacity of PLGA microspheres to form porosity in situ in injectable CPCs was optimized by investigating the influence of PLGA characteristics such as microsphere morphology (dense vs. hollow) and end-group functionalization (acid terminated vs. end-capped) on acid production and corresponding porosity formation in vitro. The current study has investigated the in vivo bone response to CPCs containing two types of microspheres (hollow and dense) made of PLGA with two different end-group functionalizations (end capped and acid terminated). Microspheres were embedded in CPC and injected in the distal femoral condyle of New Zealand White Rabbits for 6 and 12 weeks. Histological results confirmed the excellent biocompatibility and osteoconductivity of all tested materials. Composites containing acid terminated PLGA microspheres displayed considerable porosity and concomitant bone ingrowth after 6 weeks, whereas end capped microspheres only revealed open porosity after 12 weeks of implantation. In addition, it was found that dense PLGA microspheres induced significantly more CPC degradation and bone tissue formation compared to hollow PLGA microspheres. In conclusion, it was shown that PLGA microspheres have a strong capacity to induce fast degradation of injectable CPC and concomitant replacement by bone tissue by controlled release of acid polymeric degradation products without compromising the excellent biocompatibility and osteoconductivity of the CPC matrix.

In vitro degradation rate of apatitic calcium phosphate cement with incorporated PLGA microspheres

Calcium phosphate cements (CPCs) are frequently used as bone substitute material. Despite their superior clinical handling and excellent biocompatibility, they exhibit poor degradability, which limits bone ingrowth into the implant. Microspheres were prepared from poly( d, l-lactic-co-glycolic acid) (PLGA) and included in injectable CPCs as porogens in order to enhance its macroporosity after the polymeric microspheres had degraded. Upon degradation of the PLGA microspheres, acid is produced that enhances the dissolution rate of the CPC. However, the effect of the characteristics of PLGA microspheres on the degradation rate of CPCs has never been studied before. Therefore, the purpose of the current study was to investigate the dependence of CPC degradation on the chemical and morphological characteristics of incorporated PLGA microspheres. With respect to the chemical characteristics of the PLGA microspheres, the effects of both PLGA molecular weight (5, 17 and 44 kDa) and end-group functionalization (acid-terminated or end-capped) were studied. In addition, two types of PLGA microspheres, differing in morphology (hollow vs. dense), were tested. The results revealed that, although both chemical parameters clearly affected the polymer degradation rate when embedded as hollow microspheres in CPC, the PLGA and CPC degradation rates were mainly dependent on the end-group functionalization. Moreover, it was concluded that dense microspheres were more efficient porogens than hollow ones by increasing the CPC macroporosity during in vitro incubation. By combining all test parameters, it was concluded that dense PLGA microspheres consisting of acid-terminated PLGA of 17 kDa exhibited the highest and fastest acid-producing capacity and correspondingly the highest and fastest amount of porosity. In conclusion, the data presented here indicate that the combination of dense, acid-terminated PLGA microspheres with CPC emerges as a successful combination to achieve enhanced apatitic CPC degradation.

In vivo degradation of calcium phosphate cement incorporated into biodegradable microspheres

In this study we have investigated the influence of the mechanism of microsphere degradation or erosion on the in vivo degradation of microsphere/calcium phosphate cement composites (microsphere CPCs) used in tissue engineering. Microspheres composed of poly(lactic- co-glycolic acid) (PLGA), gelatin and poly(trimethylene carbonate) (PTMC) were used as the model and the resulting microsphere CPCs were implanted subcutaneously for 4, 8 or 12 weeks in the back of New Zealand white rabbits. Besides degradation, the soft tissue response to these formulations was evaluated. After retrieval, specimens were analyzed by physicochemical characterization and histological analysis. The results showed that all microsphere CPCs exhibited microsphere degradation after 12 weeks of subcutaneous implantation, which was accompanied by decreasing compression strength. The PLGA microspheres exhibited bulk erosion simultaneously throughout the whole composite, whereas the gelatin type B microspheres were degradated from the outside to the center of the composite. High molecular weight PTMC microspheres exhibited surface erosion resulting in decreasing microsphere size. Furthermore, all composites showed a similar tissue response, with decreasing capsule thickness over time and a persistent moderate inflammatory response at the implant interface. In conclusion, microsphere CPCs can be used to generate porous scaffolds in an in vivo environment after degradation of microspheres by various degradation/erosion mechanisms.