Modifications of Poly(Methyl Methacrylate) Cement for Application in Orthopedic Surgery.

Even with the emerging of newly-developed bone substitutes, poly(methyl methacrylate) (PMMA) cement is still a widely-used bone replacing biomaterial in orthopedic surgery with a long history. However, aseptic loosening, infection of the prosthesis and thermal necrosis to surrounding tissue are the common complications of PMMA. Therefore, additives have been incorporated in PMMA cement to target those problems. This chapter summarizes different additives to improve the performance of the PMMA cement, i.e.: (1) bioceramic additives; (2) filler additives; (3) antibacterial additives; (4) porogens; (5) biological agents, and (6) mixed additives. To improve the biological and mechanical performance of PMMA cement, mixed additives aiming to fabricate multifunctional PMMA seem the most suitable choice. Although in vivo animal studies have been conducted, long-term and clinical studies are still needed to evaluate the modifications of multifunctional PMMA cement for matching a specific clinical application.

Influence of polymeric additives on the cohesion and mechanical properties of calcium phosphate cements.

To expand the clinical applicability of calcium phosphate cements (CPCs) to load-bearing anatomical sites, the mechanical and setting properties of CPCs need to be improved. Specifically, organic additives need to be developed that can overcome the disintegration and brittleness of CPCs. Hence, we compared two conventional polymeric additives (i.e. carboxylmethylcellulose (CMC) and hyaluronan (HA)) with a novel organic additive that was designed to bind to calcium phosphate, i.e. hyaluronan-bisphosphonate (HABP). The unmodified cement used in this study consisted of a powder phase of α-tricalcium phosphate (α-TCP) and liquid phase of 4% NaH2PO4·2H2O, while the modified cements were fabricated by adding 0.75 or 1.5 wt% of the polymeric additive to the cement. The cohesion of α-TCP was improved considerably by the addition of CMC and HABP. None of the additives improved the compression and bending strength of the cements, but the addition of 0.75% HABP resulted into a significantly increased cement toughness as compared to the other experimental groups. The stimulatory effects of HABP on the cohesion and toughness of the cements is hypothesized to derive from the strong affinity between the polymer-grafted bisphosphonate ligands and the calcium ions in the cement matrix.

In vivo comparison between laser-treated and grit blasted/acid etched titanium.

OBJECTIVE: Laser profiling of titanium has been of considerable interest in the field of oral implantology. However, very few pre-clinical and clinical studies have been performed with laser-treated implants, especially focusing on isotropic roughness topography. The aim of the study was to compare the cortical bone response of Ti-implants discs treated with pico-sec pulsed laser (LAS) and conventional grit-blasted/acid-etched (GAE) method. MATERIALS AND METHODS: Prior to the in vivo experiment, in vitro cell viability testing of the LAS surface treatment was preformed. Then, 5 mm diameter Titanium (Ti) discs treated with LAS and GAE method were implanted in a pre-validated rabbit tibia cortical bone model and assessed with histology and histomorphometric measurements. In total, eight New Zealand White adult female rabbits were used. RESULTS: The in vitro cell viability testing with osteoblast-like cells confirmed cytocompatibility of the LAS surface treatment. Further, the rabbit experiment demonstrated a bone-to-implant contact of 68% (±17) for the laser-treated discs and 49% (±21) for the GAE discs 8 weeks after the implantation, which was statistically not different. CONCLUSION: Laser surface treatment gives the same results to the grit-blasting/acid-etched method and thus is a valid alternative to conventional roughening for dental implant materials.

Biocompatibility and bone formation with porous modified PMMA in normal and irradiated mandibular tissue.

UNLABELLED: A cemented mandibular endoprosthesis is a potentially viable option for mandibular reconstruction after ablative surgery. The commonly used PMMA cement has the inherent weakness of a lack of bioactivity. Improvement by the addition of porosities and bioactive compounds like calcium phosphates may resolve this issue. OBJECTIVE: The objective of this study was to assess the bone and tissue response to two modified PMMA cements with post-operative radiation as an additional influencing factor. MATERIALS & METHODS: An in vivo animal study was performed using a mandibular rabbit model. A porous PMMA cement (A) and a porous cement incorporated with Beta-tricalcium phosphate particles (b-TCP) (B) were placed in bilateral mandibular defects with exposed roots and mandibular nerve of 20 animals. Half of the animals underwent additional post-operative radiation. RESULTS: The animals were healthy with only a minor complication in one rabbit. Temperature analysis showed no significant risk of thermal necrosis with the maximal in vivo cement temperature at 37.8°C. Histology demonstrated: (1) good bone ingrowth around the defect as well as within the pores of the cement and defect bridging was achieved in 70% of the specimens after 12-15 weeks of implantation, (2) no pulpal injury with minor secondary cementum response, (3) an intact mandibular nerve with no inflammation, (4) extensive degradation and resorption of the b-TCP particles by 12-15 weeks, and (5) presence of an intervening thin fibrous tissue at the bone-to-cement interface. Histomorphometrical analysis revealed that there was no difference between the different cements and the presence or absence of post-operative radiation. The 12-15 weeks specimens showed significantly more bone ingrowth and bone maturity than the 4-7 weeks specimens. CONCLUSION: Both modified PMMA cements have good biocompatibility, bioactivity and support bone ingrowth and additional post-operative radiation did not show any negative effects.

Shear resistance of fiber-reinforced composite and metal dentin pins.

PURPOSE: To assess whether dentin pins increase shear resistance of extensive composite restorations and to compare performance of mini fiber-reinforced composite (FRC) anchors with metal dentin pins in the laboratory. METHODS: 30 extracted sound molars were randomly divided into three groups. Occlusal surfaces were ground flat with a standard surface area and resin composite restorations were made in Group A. In Groups B and C similar restorations were made, with additionally four metal pins placed in Group B and four FRC pins in Group C. Specimens were statically loaded until failure occurred. Failure modes were characterized as intact remaining tooth substrate (adhesive or cohesive failure of restoration) or fractured remaining tooth substrate. RESULTS: Mean failure stresses were 6.5 MPa (SD 3.2 MPa) for Group A, 9.7 MPa (SD 2.6 MPa) for Group B and 9.2 MPa (SD 2.6 MPa) for Group C. Difference in mean failure stresses between Group A and Groups B and C was statistically significant (P = 0.01), while the difference between Groups B and C was not (P = 0.63). Failures of the restoration without fracture of tooth substrate were seen for 80% of specimens in Group A and 20% in Groups B and C (P = 0.04).

Introduction of enzymatically degradable poly(trimethylene carbonate) microspheres into an injectable calcium phosphate cement.

Poly(trimethylene carbonate) (PTMC) is an enzymatically degradable polyester with rubber-like properties. Introduction of this polymer into an injectable calcium phosphate bone cement can therefore be used to introduce macroporosity into the cement for tissue engineering purposes as well as to improve mechanical properties. Aim of this study was to investigate calcium phosphate cements with incorporated PTMC microspheres (PTMC CPCs) on their physical/mechanical properties and in vitro degradation characteristics. Therefore, composites were tested on setting time and mechanical strength as well as subjected to phosphate buffered saline (PBS) and enzyme containing medium. PTMC CPCs (12.5 and 25 wt%) with molecular weights of 52.7 kg mol(-1) and 176.2 kg mol(-1) were prepared, which showed initial setting times similar to that of original CPC. Though compression strength decreased upon incorporation of PTMC microspheres, elastic properties were improved as strain-at-yield increased with increasing content of microspheres. Sustained degradation of the microspheres inside PTMC CPC occurred when incubated in the enzymatic environment, but not in PBS, which resulted in an interconnected macroporosity for the 25 wt% composites.

Bone response and mechanical strength of rabbit femoral defects filled with injectable CaP cements containing TGF-beta 1 loaded gelatin microparticles.

This study focused at the potential of transforming growth factor beta 1 (TGF-beta 1) loaded gelatin microparticles to enhance the bone response and mechanical strength of rabbit femoral defects filled with injectable calcium phosphate (CaP)/gelatin microparticle composites. Therefore, TGF-beta1 loaded composites and non-loaded controls were injected in circular defects as created in the femoral condyles of rabbits and were left in place for 4, 8 and 12 weeks. The specimens were evaluated mechanically (push-out test), and morphologically (scanning electron microscopy (SEM), histology, and histomorphometry). The results showed a gradual increase in mechanical strength with increasing implantation periods. Histological and histomorphometrical evaluation showed similar results for both composite formulations regarding histological aspect, new bone formation and bone/implant contact. However, TGF-beta1 loading of the composites demonstrated a significant effect on composite degradation after twelve weeks of implantation. The results of this study showed that CaP/gelatin composites show excellent osteogenic properties and a rapid increase in mechanical strength. The addition of TGF-beta1 significantly enhances the bone remodeling process.

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.

The cytocompatibility and early osteogenic characteristics of an injectable calcium phosphate cement.

In this study, the cytocompatibility and early osteogenic characteristics of rat bone marrow cells (RBMCs) on injectable calcium phosphate (CaP) cement (Calcibon) were investigated. In addition to unmodified CaP cement discs, 2 other treatments were given to the discs: preincubation in MilliQ and sintering at different temperatures. After primary culture, RBMCs were dropwise seeded on the discs and cultured for 12 days. The samples were evaluated in terms of cell viability, morphology (live and dead assays and scanning electron microscopy (SEM)), cell proliferation (deoxyribonucleic acid (DNA) analyses), early cell differentiation (alkaline phosphatase (ALP) activity), and physicochemical analyses (x-ray diffraction (XRD)). The live and dead, DNA, and SEM results showed that Calcibon discs without any additional treatment were not supporting osteoblast-like cells in vitro. There were fewer cells, and cell layers were detached from the disc surface. Therefore, different preincubation periods and sintering temperatures were evaluated to improve the cytocompatibility of the CaP cement. Preincubating discs in MilliQ for periods of 1, 4, 8, and 12 weeks resulted in the hydrolysis of alpha-tri calcium phosphate (TCP) into an apatite-like structure with some beta-TCP, as shown with XRD, but the material was not cytocompatible. Sintering the discs between 800 degrees C and 1100 degrees C resulted in conversion of alpha-TCP to beta-TCP with some hydroxyapatite and an increase in crystallinity. Eventually, the discs sintered at 1100 degrees C achieved better cell attachment, more-abundant cell proliferation, and earlier differentiation than other sintered (600 degrees C, 800 degrees C, and 1000 degrees C), preincubated, and unmodified specimens. On basis of our results, we conclude that in vivo results with CaP-based cements do not guarantee in vitro applicability. Furthermore, unmodified Calcibon is not cytocompatible in vitro, although preincubation of the material results in a more-favorable cell response, sintering of the material at 1100 degrees C results in the best osteogenic properties. In contrast to in vivo studies, the Calcibon CaP cement is not suitable as a scaffold for cell-based tissue-engineering strategies.

Bone regeneration of porous beta-tricalcium phosphate (Conduit TCP) and of biphasic calcium phosphate ceramic (Biosel) in trabecular defects in sheep.

In this study bone regeneration between porous beta-tricalcium phosphate (Conduit TCP) and biphasic calcium phosphate ceramic (Biosel), with a hydroxyapatite/beta-TCP ratio of 75/25, was compared. The ceramic particles were implanted in sheep trabecular bone for 3, 12, and 26 weeks. Histomorphometrical analysis revealed that Conduit degraded significantly during time and only 36% of the material was left at 26 weeks implantation time. Biosel, in contrast, remained nearly intact. The degradation of Conduit was due to dissolution as well as cell-mediated. Biosel showed a high cellular intervention, although this material did not degrade. Both materials were osteoconductive. The amount of newly formed bone appeared greater in the Conduit group after 26 weeks (46% +/- 8% as compared to 37% +/- 8% for Biosel), but this difference was not significant. Bone distribution over the defect was homogeneous in Conduit, whereas Biosel showed significantly more bone in the periphery of the defect after 26 weeks in comparison to the center. In conclusion, both ceramics are biocompatible and osteoconductive. Degradation showed a difference in amount and in cellular events, with more degraded Conduit TCP with less cellular intervention as compared to Biosel.

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.

The effectiveness of different polymerization protocols for class II composite resin restorations.

OBJECTIVES: To investigate the effect of reduced light exposure times on Vickers hardness (VH) of class II composite resin restorations. METHODS: Class II restorations were made in vitro in three 2mm thick increments in a human molar. Two composite resins (Clearfil AP-X; Esthet-X) were polymerized with four light-curing units (Halogen; Astralis 10, LED; The Cure, L.E. Demetron I, Smartlite) following four curing protocols. Three protocols with exposure times of 10s, 20s or 40s (control) per layer. In the fourth protocol, 10s irradiation per layer was combined with additional lateral curing for 10s from buccal and palatal after removal of the metal matrix. VH of the axial surface was determined at top and bottom layers directly after light-curing and after 7 days storage. Linear regression analysis was performed to analyze the effect of protocol variables. RESULTS: Directly after light-curing VH of both composite resins was significantly influenced by curing protocols. After 7 days, curing protocols had no significant effect on VH of Clearfil AP-X, except for the Smartlite. VH of Esthet-X was still influenced by curing protocol, but differences were smaller than directly after light-curing. CONCLUSIONS: With high intensity light-curing units, exposure times of 10s/2mm increment can be sufficient to obtain under in vitro conditions a high degree of conversion, depending on materials and curing protocols. With additional lateral curing of a class II composite resin restoration a higher degree of cure can be obtained in less time.

Bone Response to Porous Poly(methyl methacrylate) Cement Loaded with Hydroxyapatite Particles in a Rabbit Mandibular Model.

The aim of the current study was to evaluate bone formation and tissue response to porous poly(methyl methacrylate) (PMMA) cement with or without hydroxyapatite (HA) in a rabbit mandibular model. Therefore, 14 New Zealand White rabbits were randomly divided into two groups of seven according to the designed study end points of 4 and 12 weeks. For each rabbit, two decorticated defects (6 mm in height and 10 mm in width for each) were prepared at both sides of the mandible. Subsequently, the defects were filled with, respectively, porous PMMA and porous PMMA-HA cement. After reaching the designated implantation period, the rabbits were euthanized and the mandibles were retrieved for histological analysis. Results showed that both porous PMMA and porous PMMA-HA supported bone repair. Neither of the bone cements caused significant inflammation to nerve or other surrounding tissues. After implantation of 12 weeks, majority of the porosity was filled with newly formed bone for both cements, which supports the concept that a porous structure within PMMA can enhance bone ingrowth. Histomorphometrical evaluation, using histological grading scales, demonstrated that, at both implantation times, the presence of HA in the PMMA enhanced bone formation. Bone was always in direct contact with the HA particles, while intervening fibrous tissue was present at the PMMA-bone interface. On the basis of results, it was concluded that injectable porous PMMA-HA cement might be a good candidate for craniofacial bone repair, which should be further evaluated in a more clinically relevant large animal model.

In vitro and in vivo reactivity of porous, electrosprayed calcium phosphate coatings.

The dissolution and/or precipitation behaviour of porous calcium phosphate (CaP) coatings, deposited using electrostatic spray deposition (ESD), was investigated (a) in vitro after soaking in simulated body fluid (SBF) for several time periods (2, 4, 8, and 12 weeks), and (b) in vivo after subcutaneous implantation of CaP-coated implants in the back of goats for identical time periods. Physical and chemical properties of coatings were characterized before and after in vitro/vivo testing by means of scanning electron microscopy, X-ray diffraction, Fourier-transform infrared spectroscopy, and energy dispersive spectroscopy. Moreover, part of the explants was prepared for light microscopical evaluation of the tissue response. In vitro, all apatitic ESD-coatings induced the formation of homogeneous and adherent CaP precipitation layers. Amorphous CaP, however, displayed a delayed precipitation of poorly adherent CaP layers, whereas heterogeneous calcification was observed on top of beta-TCP-coated substrates, indicating that beta-TCP and amorphous CaP coatings exhibit a poor ability of inducing calcification in SBF as compared to crystalline apatitic coatings. In vivo, no adverse tissue reactions (toxic effects/inflammatory cells) were observed using light microscopy, and all coatings became surrounded by a dense, fibrous tissue capsule after implantation. All ESD-coatings degraded gradually at a dissolution rate depending on the chemical phase (order of relative solubility: amorphous CaP approximately carbonate apatite>beta-TCP>carbonated hydroxyapatite), thereby enabling synthesis of CaP coatings with a tailored degradation rate.

Mechanical evaluation of implanted calcium phosphate cement incorporated with PLGA microparticles.

In this study, the mechanical properties of an implanted calcium phosphate (CaP) cement incorporated with 20wt% poly (dl-lactic-co-glycolic acid) (PLGA) microparticles were investigated in a rat cranial defect. After 2, 4 and 8 weeks of implantation, implants were evaluated mechanically (push-out test) and morphologically (Scanning Electron Microscopy (SEM) and histology). The results of the push-out test showed that after 2 weeks the shear strength of the implants was 0.44+/-0.44MPa (average+/-sd), which increased to 1.34+/-1.05MPa at 4 weeks and finally resulted in 2.60+/-2.78MPa at 8 weeks. SEM examination showed a fracture plane at the bone-cement interface at 2 weeks, while the 4- and 8-week specimens created a fracture plane into the CaP/PLGA composites, indicating an increased strength of the bone-cement interface. Histological evaluation revealed that the two weeks implantation period resulted in minimal bone ingrowth, while at 4 weeks of implantation the peripheral PLGA microparticles were degraded and replaced by deposition of newly formed bone. Finally, after 8 weeks of implantation the degradation of the PLGA microparticles was almost completed, which was observed by the bone ingrowth throughout the CaP/PLGA composites. On basis of our results, we conclude that the shear strength of the bone-cement interface increased over time due to bone ingrowth into the CaP/PLGA composites. Although the bone-cement contact could be optimized with an injectable CaP cement to enhance bone ingrowth, still the mechanical properties of the composites after 8 weeks of implantation are insufficient for load-bearing purposes.

The effect of platelet-rich plasma on the bone healing around calcium phosphate-coated and non-coated oral implants in trabecular bone.

The effect of local application of autologous platelet-rich plasma (PRP) on bone healing in combination with the use of titanium implants with 2 different surface configurations was investigated. PRP fractions were obtained from venous blood sample of 6 goats and applied via gel preparation and subsequent installation in the implant site or via dipping of the implant in PRP liquid before insertion. Thirty-six implants (18 non-coated and 18 calcium phosphate (CaP) coated) were placed into the goat femoral condyles (trabecular bone). The animals were sacrificed at 6 weeks after implantation, and implants with surrounding tissue were processed for light microscopical evaluation. In addition to subjective description of the histological findings, histomorphometrical variables were also evaluated (the bone-implant contact and the bone mass adjacent to the implant). Significantly more interfacial bone-to-implant contact was observed for all 3 groups of CaP-coated implants and the titanium / liquid group (non-coated implant with PRP liquid) than for the other 2 non-coated titanium groups (with PRP gel or without PRP). The evaluation of the bone mass close to implant surface indicated that all the groups induced a significant increase of the bone mass except the PRP gel groups. On the basis of the observations, it was concluded that magnetron-sputtered CaP coatings can improve the integration of oral implants in trabecular bone. The additional use of PRP did not offer any significant effect on the bone response to the CaP-coated implants, whereas PRP in a liquid form showed a significant effect on bone apposition to roughened titanium implants during the early post-implantation healing phase.

Transforming growth factor-beta1 release from a porous electrostatic spray deposition-derived calcium phosphate coating.

This study evaluated the utilization of a porous coating, derived with electrostatic spray deposition (ESD), as a carrier material for transforming growth factor-beta1 (TGF-beta1). A porous beta-tricalcium phosphate coating was deposited with ESD, and 10 ng of (125) I-labeled TGF-beta1 was loaded on the substrates. A burst release during the first hour of incubation of >90% was observed, in either culture medium or phosphate-buffered saline (PBS). Ninety-nine percent of the growth factor was released after 10 days of incubation. All samples were able to inhibit epithelial cell growth, indicating that the growth factor had remained bioactive after release. Thereafter, osteoblast-like cells were seeded upon substrates with or without 10 ng of TGF-beta1. While proliferation of osteoblast-like cells was increased on TGF-beta1-loaded substrates, differentiation was inhibited or delayed. In conclusion, a porous ESD-derived calcium phosphate coating can be used as a carrier material for TGF-beta1, when a burst release is desired.

Physicochemical properties and mineralization assessment of porous polymethylmethacrylate cement loaded with hydroxyapatite in simulated body fluid.

The aim of this study was to evaluate the effect of carboxymethylcellulose (CMC) as a pore generator and hydroxyapatite (HA) as an osteoconductive agent on the physicochemical properties and in-vitro mineralization ability of porous polymethylmethacrylate (PMMA) cement. To this end, various compositions of PMMA cements, which differed in amount of millimeter-sized hydroxyapatite (HA) particles and CMC hydrogel, were prepared and immersed into simulated body fluid (SBF) for 0, 7, 14, 21 and 28 days. It was demonstrated that the incorporation of CMC hydrogel decreased the maximum temperature of cement to the normal body temperature and prolonged the handling time during polymerization. Further, the amount of CMC was responsible for the creation of porosity and interconnectivity, which in turn determined the final mechanical properties of cements. The loaded HA particles enhanced the potential bioactivity of cement for bone ingrowth. Albeit different amount of HA particles influenced their final exposures on the surface of cured cement, all of the three amounts of HA did not weaken the final mechanical properties of cements. The data here suggests that the HA particle loaded porous PMMA cement can serve as the promising candidate for bone reconstruction.

Effect of calcium phosphate ceramic substrate geometry on mesenchymal stromal cell organization and osteogenic differentiation.

The composition of calcium phosphate (CaP) ceramics in combination with surface features have been shown to influence biological performance, and micro- and nano-scale topography is known to stimulate osteogenic differentiation of mesenchymal stromal cells (MSCs). In view of this, adipose tissue derived MSCs were cultured on CaP disks featuring hemispherical concavities of various sizes (440, 800 or 1800 µm diameter). It was hypothesized that (i) surface concavities would promote cell proliferation, cellular organization within the concavities, and osteogenic differentiation, as a result of a more pronounced 3D micro-environment and CaP nucleation in concavities, and (ii) MSC proliferation and osteogenic differentiation would increase with smaller concavity size due to more rapidly occurring 3D cell-cell interactions. We found that concavities indeed affect cell proliferation, with 440 µm concavities increasing cell proliferation to a larger extent compared to 800 and 1800 µm concavities as well as planar surfaces. Additionally, concavity size influenced 3D cellular organization within the concavity volume. Interestingly, concavity size promoted osteogenic differentiation of cells, as evidenced by increased osteocalcin gene expression in 440 µm concavities, and osteocalcin staining predominantly for 440 and 800 µm concavities, but not for 1800 µm concavities and only slightly for planar surface controls.

Size matters: effects of PLGA-microsphere size in injectable CPC/PLGA on bone formation.

The aim of this study was to evaluate the effect of PLGA microsphere dimensions on bone formation after injection of calcium phosphate cement (CPC)/PLGA in a guinea pig tibial intramedullarly model. To this end, injectable CPC/PLGA formulations were prepared using PLGA microspheres with either a small (~25 µm) or large (~100 µm) diameter, which were incorporated at a 20:80 ratio (wt%) within apatite CPC. Both CPC/PLGA formulations were injected into a marrow-ablated tibial intramedullary cavity and, after an implantation period of 12 weeks, histology and histomorphometry were used to address bone formation. The results demonstrated bone ingrowth throughout the entire scaffold material for both CPC/PLGA formulations upon PLGA microsphere degradation. More importantly, bone formation within the CPC matrix was > two-fold higher for CPC-PLGA with 25 µm PLGA microspheres. Additionally, the pattern of bone and marrow formation showed distinct differences related to PLGA microsphere dimension. In general, this study demonstrates that PLGA microsphere dimensions of ~25 µm, leading to pores of ~25 µm within CPC, are sufficient for bone ingrowth and allow substantial bone formation. Further, the results demonstrate that PLGA microsphere dimensions provide a tool to control bone formation for injectable CPC/PLGA bone substitutes. Copyright © 2013 John Wiley & Sons, Ltd.