A systematic review on the long-term success of calcium phosphate plasma-spray-coated dental implants.

The objectives of the current review were (1) to systematically appraise, and (2) to evaluate long-term success data of calcium phosphate (CaP) plasma-spray-coated dental implants in clinical trials with at least 5 years of follow-up. To describe the long-term efficacy of functional implants, the outcome variables were (a) percentage annual complication rate (ACR) and (b) cumulative success rate (CSR), as presented in the selected articles. The electronic search yielded 645 titles. On the basis of the inclusion criteria, 8 studies were finally included. The percentage of implants in function after the first year was estimated to be 98.4 % in the maxilla and 99.2 % in the mandible. The estimates of the weighted mean ACR-percentage increased over the years up to 2.6 (SE 0.7) during the fifth year of function for the maxilla and to 9.4 (SE 8.4) for the mandible in the tenth year of function. After 10 years, the mean percentage of successful implants was estimated to be 71.1 % in the maxilla and 72.2 % in the mandible. The estimates seem to confirm the proposed, long-term progressive bone loss pattern of CaP-ceramic-coated dental implants. Within the limits of this meta-analytic approach to the literature, we conclude that: (1) published long-term success data for calcium phosphate plasma-spray-coated dental implants are limited, (2) comparison of the data is difficult due to differences in success criteria among the studies, and (3) long-term CSRs demonstrate very weak evidence for progressive complications around calcium phosphate plasma-spray-coated dental implants.

Influence of surface microstructure and chemistry on osteoinduction and osteoclastogenesis by biphasic calcium phosphate discs.

It has been reported that surface microstructural dimensions can influence the osteoinductivity of calcium phosphates (CaPs), and osteoclasts may play a role in this process. We hypothesised that surface structural dimensions of ≤ 1 µm trigger osteoinduction and osteoclast formation irrespective of macrostructure (e.g., concavities, interconnected macropores, interparticle space) or surface chemistry. To test this, planar discs made of biphasic calcium phosphate (BCP: 80% hydroxyapatite, 20% tricalcium phosphate) were prepared with different surface structural dimensions - either ~ 1 µm (BCP1150) or ~ 2-4 µm (BCP1300) - and no macropores or concavities. A third material was made by sputter coating BCP1150 with titanium (BCP1150Ti), thereby changing its surface chemistry but preserving its surface structure and chemical reactivity. After intramuscular implantation in 5 dogs for 12 weeks, BCP1150 formed ectopic bone in 4 out of 5 samples, BCP1150Ti formed ectopic bone in 3 out of 5 samples, and BCP1300 formed no ectopic bone in any of the 5 samples. In vivo, large multinucleated osteoclast-like cells densely colonised BCP1150, smaller osteoclast-like cells formed on BCP1150Ti, and osteoclast-like cells scarcely formed on BCP1300. In vitro, RAW264.7 cells cultured on the surface of BCP1150 and BCP1150Ti in the presence of osteoclast differentiation factor RANKL (receptor activator for NF-κB ligand) proliferated then differentiated into multinucleated osteoclast-like cells with positive tartrate resistant acid phosphatase (TRAP) activity. However, cell proliferation, fusion, and TRAP activity were all significantly inhibited on BCP1300. These results indicate that of the material parameters tested - namely, surface microstructure, macrostructure, and surface chemistry - microstructural dimensions are critical in promoting osteoclastogenesis and triggering ectopic bone formation.

Osteochondral defect repair using bilayered hydrogels encapsulating both chondrogenically and osteogenically pre-differentiated mesenchymal stem cells in a rabbit model.

OBJECTIVE: To investigate the ability of cell-laden bilayered hydrogels encapsulating chondrogenically and osteogenically (OS) pre-differentiated mesenchymal stem cells (MSCs) to effect osteochondral defect repair in a rabbit model. By varying the period of chondrogenic pre-differentiation from 7 (CG7) to 14 days (CG14), the effect of chondrogenic differentiation stage on osteochondral tissue repair was also investigated. METHODS: Rabbit MSCs were subjected to either chondrogenic or osteogenic pre-differentiation, encapsulated within respective chondral/subchondral layers of a bilayered hydrogel construct, and then implanted into femoral condyle osteochondral defects. Rabbits were randomized into one of four groups (MSC/MSC, MSC/OS, CG7/OS, and CG14/OS; chondral/subchondral) and received two similar constructs bilaterally. Defects were evaluated after 12 weeks. RESULTS: All groups exhibited similar overall neo-tissue filling. The delivery of OS cells when compared to undifferentiated MSCs in the subchondral construct layer resulted in improvements in neo-cartilage thickness and regularity. However, the addition of CG cells in the chondral layer, with OS cells in the subchondral layer, did not augment tissue repair as influenced by the latter when compared to the control. Instead, CG7/OS implants resulted in more irregular neo-tissue surfaces when compared to MSC/OS implants. Notably, the delivery of CG7 cells, when compared to CG14 cells, with OS cells stimulated morphologically superior cartilage repair. However, neither osteogenic nor chondrogenic pre-differentiation affected detectable changes in subchondral tissue repair. CONCLUSIONS: Cartilage regeneration in osteochondral defects can be enhanced by MSCs that are chondrogenically and osteogenically pre-differentiated prior to implantation. Longer chondrogenic pre-differentiation periods, however, lead to diminished cartilage repair.

Maxillary sinus augmentation with microstructured tricalcium phosphate ceramic in sheep.

OBJECTIVE: The objective of this study was to evaluate the biological performance of osteoinductive microstructured tricalcium phosphate (MSTCP) particles in maxillary sinus floor augmentation surgery in sheep. MATERIAL AND METHODS: Sinus floor augmentation was performed in eight Swifter sheep. In each animal, the maxillary sinus floor was unilaterally augmented with MSTCP particles. Computed tomography (CT) imaging and histological analyses were performed after 12 weeks of implantation. RESULTS: Maxillofacial CT, histology, histomorphometrical analysis and sequential polychrome fluorescent labeling indicated that MSTCP particles provided a scaffold for cell ingrowth and bone formation. After a 12-week implantation period, the sinuses grafted with MSTCP showed an increased bone height of 6 mm and a mean total bone volume of 43%, with significant degradation of MSTCP particles. CONCLUSION: MSTCP particles represent a suitable bone substitute material for maxillary sinus floor augmentation surgery.

Predictive value of ridge dimensions on autologous bone graft resorption in staged maxillary sinus augmentation surgery using Cone-Beam CT.

INTRODUCTION: No studies are available that provide predictive parameters regarding the expected amount of resorption after maxillary sinus augmentation surgery using autologous bone grafts. Therefore, the aim of this study was to determine parameters influencing the outcome of the bone graft resorption process. MATERIAL AND METHODS: In 20 patients, three-dimensional analysis of alveolar ridge dimensions and bone graft volume change in the atrophic posterior maxilla was performed by Cone-Beam Computerized Tomography imaging. Ridge dimensions were assessed before maxillary sinus augmentation surgery. Bone graft volumes were compared after maxillary sinus floor augmentation surgery and a graft healing interval of several months. To analyze the relation between bone volume changes with the independent variables, patients' gender, age, alveolar crest height and width, and graft healing time interval, a multi-level extension of linear regression was applied. RESULTS: A residual bone height of 6.0 mm (SD = 3.6 mm) and 6.2 mm (SD = 3.6 mm) was found at the left and right sides, respectively. Moreover, alveolar bone widths of 6.5 mm (SD = 2.2 mm) and 7.0 mm (SD = 2.3 mm) at the premolars, and 8.8 mm (SD = 2.2 mm) and 8.9 mm (SD = 2.5 mm) at the molars regions were found at the left and right site, respectively. Bone graft volume decreased by 25.0% (SD = 21.0%) after 4.7 months (SD = 2.7, median = 4.0 months) of healing time. The variables "age" (P = 0.009) and mean alveolar crest "bone height" (P = 0.043), showed a significant influence on bone graft resorption. A decrease of 1.0% (SE = 0.3%) of bone graft resorption was found for each year the patient grew older, and an increase in bone graft resorption of 1.8% (SE = 0.8%) was found for each mm of original bone height before sinus floor augmentation. CONCLUSIONS: Graft resorption occurs when using autologous bone grafts for maxillary sinus augmentation. Alveolar crest bone height and patient age have a significant effect on graft resorption, with increased resorption for higher alveolar crest bone height and decreased resorption for older patients. Consequently, patient characteristics that affect the process of bone graft resorption should be given full consideration, when performing sinus augmentation surgery.

Biomimetic modification of synthetic hydrogels by incorporation of adhesive peptides and calcium phosphate nanoparticles: in vitro evaluation of cell behavior.

The ultimate goal of this work was to develop a biocompatible and biomimetic in situ crosslinkable hydrogel scaffold with an instructive capacity for bone regenerative treatment. To this end, synthetic hydrogels were functionalized with two key components of the extracellular matrix of native bone tissue, i.e. the three-amino acid peptide sequence RGD (which is the principal integrin-binding domain responsible for cell adhesion and survival of anchorage-dependent cells) and calcium phosphate (CaP) nanoparticles in the form of hydroxyapatite (which are similar to the inorganic phase of bone tissue). Rat bone marrow osteoblast-like cells (OBLCs) were encapsulated in four different biomaterials (plain oligo(poly(ethylene glycol) fumarate) (OPF), RGD-modified OPF, OPF enriched with CaP nanoparticles and RGD-modified OPF enriched with CaP nanoparticles) and cell survival, cell spreading, proliferation and mineralized matrix formation were determined via cell viability assay, histology and biochemical analysis for alkaline phosphatase activity and calcium. This study showed that RGD peptide sequences promoted cell spreading in OPF hydrogels and hence play a crucial role in cell survival during the early stage of culture, whereas CaP nanoparticles significantly enhanced cell-mediated hydrogel mineralization. Although cell spreading and proliferation activity were inhibited, the combined effect of RGD peptide sequences and CaP nanoparticles within OPF hydrogel systems elicited a better biological response than that of the individual components. Specifically, both a sustained cell viability and mineralized matrix production mediated by encapsulated OBLCs were observed within these novel biomimetic composite systems.

Calcium phosphate cements: Optimization toward biodegradability.

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

Bone tumor-targeted delivery of theranostic 195mPt-bisphosphonate complexes promotes killing of metastatic tumor cells.

Platinum-based drugs such as cisplatin are very potent chemotherapeutics, whereas radioactive platinum (195mPt) is a rich source of low-energy Auger electrons, which kills tumor cells by damaging DNA. Auger electrons damage cells over a very short range. Consequently, 195mPt-based radiopharmaceuticals should be targeted toward â€‹tumors to maximize radiotherapeutic efficacy and minimize Pt-based systemic toxicity. Herein, we show that systemically administered radioactive bisphosphonate-functionalized platinum (195mPt-BP) complexes specifically accumulate in intratibial bone metastatic lesions in mice. The 195mPt-BP complexes accumulate 7.3-fold more effectively in bone 7 days after systemic delivery compared to 195mPt-cisplatin lacking bone-targeting bisphosphonate ligands. Therapeutically, 195mPt-BP treatment causes 4.5-fold more γ-H2AX formation, a biomarker for DNA damage in metastatic tumor cells compared to 195mPt-cisplatin. We show that systemically administered 195mPt-BP is radiotherapeutically active, as evidenced by an 11-fold increased DNA damage in metastatic tumor cells compared to non-radioactive Pt-BP controls. Moreover, apoptosis in metastatic tumor cells is enhanced more than 3.4-fold upon systemic administration of 195mPt-BP vs. radioactive 195mPt-cisplatin or non-radioactive Pt-BP controls. These results provide the first preclinical evidence for specific accumulation and strong radiotherapeutic activity of 195mPt-BP in bone metastatic lesions, which offers new avenues of research on radiotherapeutic killing of tumor cells in bone metastases by Auger electrons.

A novel implantation model for evaluation of bone healing response to dental implants: the goat iliac crest.

OBJECTIVES: Despite the availability of numerous animal models for testing the biological performance of dental and orthopedic implants, the selection of a suitable model is complex. This paper presents a new model for objective and standardized evaluation of bone responses to implants using the iliac crest in goats. MATERIAL AND METHODS: The feasibility of the iliac crest model regarding anatomy and implant positioning was determined using two cadaveric specimens and the bone structure was evaluated and compared with that of the goat femoral condyle. Additionally, the validity of the model was tested by performing an in vivo study. RESULTS: By means of a rather simple, safe, fast and reproducible surgical procedure, the iliac crest in goats could be approached and allowed the implantation of maximally five dental implants per iliac crest. Because of the bilateral implantation possibility, statistical comparisons between groups on either side of the goat could be performed, resulting in a high statistical power, and hence a reduction in the number of animals required to obtain significant data. CONCLUSIONS: In terms of surgical approach, anatomy and implant positioning, the iliac crest is the preferred model over the femoral condyle model. The iliac crest implantation model is suitable for evaluation of the osteogenic response to bone implant materials and represents a justified and deliberate alternative to the already existing animal models.

Effects of implant geometry, surface properties, and TGF-beta1 on peri-implant bone response: an experimental study in goats.

OBJECTIVES: Despite the high success rates in implantology, the desire to use oral implants in more challenging clinical situations drives the need for continuing refinements in implant design and surface properties. In the present study, the effect of implant geometry on implant bone response was evaluated using two geometrically different implant types, i.e. screw type (St) and push-in type(Pi). Furthermore, the potential beneficial effect of an electrosprayed calcium phosphate (CaP) coating, either or not enriched with the osteoinductive growth factor TGF-beta1, on the osteogenic response was examined. MATERIAL AND METHODS: A total of 54 implants, divided into six groups (n=9), were inserted into the femoral condyles of nine goats. After an implantation period of 12 weeks, retrieved specimens were evaluated histologically and histomorphometrically. Measurements were statistically evaluated using SPSS 14.0 and analyzed using a linear regression model. RESULTS: With respect to implant design, St-implants showed an overall superior biological healing response compared with Pi-implants. Considering surface properties, the deposition of an electrosprayed CaP (2-3 microm) coating onto implants significantly increased the amount of bone-implant contact for both implant types. Additional enrichment of the CaP coating with the osteoinductive growth factor TGF-beta1 did not significantly affect peri-implant bone response. CONCLUSIONS: The results of this study indicate that a substantial improvement of the osteogenic response to titanium implants can be achieved by the deposition of an electrosprayed CaP coating. The enrichment of the coating with 1 microg TGF-beta1 has only a marginal effect.

Complement proteins regulating macrophage polarisation on biomaterials.

One of the events occurring when a biomaterial is implanted in an host is the protein deposition onto its surface, which might regulate cell responses. When a biomaterial displays a compromised biocompatibility, distinct complement pathways can be activated to produce a foreign body reaction. In this article, we have designed different types of biomaterial surfaces to study the inflammation process. Here, we used different concentrations of (3-glycidoxypropyl)-trimethoxysilane (GPTMS), an organically-modified alkoxysilane as a precursor for the synthesis of various types of sol-gel materials functionalizing coatings for titanium implants to regulate biological responses. Our results showed that greater GPTMS surface concentrations induced greater secretion of TNF-α and IL-10 on RAW 264.7 macrophages. When implanted into rabbit tibia, osseointegration decreased with higher GPTMS concentrations. Interestingly, higher deposition of complement-related proteins C-reactive protein (CRP) and ficolin-2 (FCN2), two main activators of distinct complement pathways, was observed. Taking all together, inflammatory potential increase seems to be GPTMS concentration-dependent. Our results show that a greater adsorption of complement proteins can condition macrophage polarization.

Comparison of a resorbable magnesium implant in small and large growing-animal models.

Fracture treatment in children needs new implant materials to overcome disadvantages associated with removal surgery. Magnesium-based implants constitute a biocompatible and bioresorbable alternative. In adults and especially in children, implant safety needs to be evaluated. In children the bone turnover rate is higher and implant material might influence growth capacity, and the long-term effect of accumulated particles or ions is more critical due to the host's prolonged post-surgery lifespan. In this study we aimed to investigate the degradation behavior of ZX00 (Mg-0.45Zn-0.45Ca; in wt.%) in a small and a large animal model to find out whether there is a difference between the two models (i) in degradation rate and (ii) in bone formation and in-growth. Our results 6, 12 and 24 weeks after ZX00 implantation showed no negative effects on bone formation and in-growth, and no adverse effects such as fibrotic or sclerotic encapsulation. The degradation rate did not significantly differ between the two growing-animal models, and both showed slow and homogeneous degradation performance. Our conclusion is that small animal models may be sufficient to investigate degradation rates and provide preliminary evidence on bone formation and in-growth of implant materials in a growing-animal model. STATEMENT OF SIGNIFICANCE: The safety of implant material is of the utmost importance, especially in children, who have enhanced bone turnover, more growth capacity and longer postoperative lifespans. Magnesium (Mg)-based implants have long been of great interest in pediatric orthopedic and trauma surgery, due to their good biocompatibility, biodegradability and biomechanics. In the study documented in this manuscript we investigated Mg-Zn-Ca implant material without rare-earth elements, and compared its outcome in a small and a large growing-animal model. In both models we observed bone formation and in-growth which featured no adverse effects such as fibrotic or sclerotic encapsulation, and slow homogeneous degradation performance of the Mg-based implant material.

Macrophage behavior on multilayered DNA-coatings in vitro.

A pivotal factor to consider in the development of biomaterials and biomaterial coatings is the inflammatory response to these materials. The insertion of implants is followed by protein adsorption and subsequent interactions with cellular components of the biological surroundings, in which macrophages play a dominant role through the production of a myriad of signaling molecules. In view of this, the aims of the present study were to evaluate (i) gross protein adsorption to, and (ii) in vitro behavior of macrophages on novel biomaterial coatings, composed of poly-D-lysine (PDL) or poly(allylamine hydrochloride) (PAH) and DNA, and to compare these coatings with negative (noncoated glass) and positive controls (noncoated glass + LPS-stimulation). The results demonstrate that multilayered DNA-coatings do not affect gross protein adsorption compared to noncoated controls. Cell culture experiments showed that the attachment to, and viability and morphology of two types of macrophages cultured on multilayered DNA-coatings is comparable to noncoated controls. Still, macrophages repeatedly showed decreased secretion levels of the proinflammatory cytokine TNF-alpha on multilayered DNA-coatings, whereas no differences were observed in the secretion of IL-1beta, IL-10, and TGF-beta1. Appropriate animal studies are required to elucidate if these in vitro indications have clinical effects on the inflammatory and wound healing processes around implants.

Multilayered DNA coatings: in vitro bioactivity studies and effects on osteoblast-like cell behavior.

This study describes the effect of multilayered DNA coatings on (i) the formation of mineralized depositions from simulated body fluids (SBF); and (ii) osteoblast-like cell behavior with and without pretreatment in SBF. DNA coatings were generated using electrostatic self-assembly, with poly-d-lysine or poly(allylamine hydrochloride) as cationic polyelectrolytes, on titanium substrates. Coated substrates and non-coated controls were immersed in SBF with various compositions. The deposition of calcium phosphate was enhanced on multilayered DNA coatings as compared with non-coated controls, and was dependent on the type of cationic polyelectrolyte used in the build-up of the DNA coatings. Further analysis showed that the depositions consisted of carbonated apatite. Non-pretreated DNA coatings were found to have no effect on osteoblast-like cell behavior compared with titanium controls. On the other hand, SBF-pretreatment of DNA coatings affected the differentiation of osteoblast-like cells through an increased deposition of osteocalcin. The results of this study are indicative of the bone-bonding capacities of DNA coatings. Nevertheless, future animal experiments are required to provide conclusive evidence for the bioactivity of DNA coatings.

Biological responses to multilayered DNA-coatings.

This study was performed to evaluate the basic biological response to deoxyribonucleic acid (DNA)-based coatings for soft tissue implants. To that end, in vitro experiments were used to study their cytocompatibility, and in vivo subcutaneous implantation studies with transponders in a rat model were performed to evaluate their histocompatibility. The DNA-based coatings were fabricated using the electrostatic self-assembly technique using cationic poly-D-lysine or poly-allylamine hydrochloride and anionic DNA. Noncoated substrates served as controls. In vitro, the behavior of primary rat dermal fibroblasts was assessed in terms of cell proliferation and morphology. Both types of multilayered DNA-coatings significantly increased rat dermal fibroblast proliferation without altering the morphological appearance of the cells. The tissue response to multilayered DNA-coatings was assessed using an in vivo rat model, in which transponders were inserted subcutaneously for 4 and 12 weeks. No macroscopic signs of inflammation or adverse tissue reactions were observed at implant retrieval. Histological analyses demonstrated a uniform tissue response to all types of implants. All implants were encapsulated in a fibrous tissue capsule without intervening inflammatory cells at the implant surface. Histomorphometrically, multilayered DNA-coatings induced fibrous tissue capsules with similar quality and thickness compared to noncoated controls. In addition, all fibrous tissue capsules showed similar expression of alpha-smooth muscle actin. This study demonstrates that multilayered DNA-coatings are cytocompatible and histocompatible, and justifies further research on their functionalization with biologically active compounds to modulate tissue responses.

Porous titanium scaffolds with injectable hyaluronic acid-DBM gel for bone substitution in a rat critical-sized calvarial defect model.

Demineralized bone matrix (DBM) is an allograft bone substitute used for bone repair surgery to overcome drawbacks of autologous bone grafting, such as limited supply and donor-site comorbidities. In view of different demineralization treatments to obtain DBM, we examined the biological performance of two differently demineralized types of DBM, i.e. by acidic treatment using hydrochloric acid (HCl) or treatment with the chelating agent ethylene diamine tetra-acetate (EDTA). First, we evaluated the osteo-inductive properties of both DBMs by implanting the materials subcutaneously in rats. Second, we evaluated the effects on bone formation by incorporating DBM in a hyaluronic acid (HA) gel to fill a porous titanium scaffold for use in a critical-sized calvarial defect model in 36 male Wistar rats. These porous titanium scaffolds were implanted empty or filled with HA gel containing either DBM HCl or DBM EDTA. Ectopically implanted DBM HCl and DBM EDTA did not induce ectopic bone formation over the course of 12 weeks. For the calvarial defects, mean percentages of newly formed bone at 2 weeks were significantly higher for Ti-Empty compared to Ti-HA + DBM HCl, but not compared to Ti-HA + DBM EDTA. Significant temporal bone formation was observed for Ti-Empty and Ti-HA + DBM HCl, but not for Ti-HA + DBM EDTA. At 8 weeks there were no significant differences in values of bone formation between the three experimental constructs. In conclusion, these results showed that, under the current experimental conditions, neither DBM HCl nor DBM EDTA possess osteo-inductive properties. Additionally, in combination with an HA gel loaded in a porous titanium scaffold, DBM HCl and DBM EDTA showed similar amounts of new bone formation after 8 weeks, which were lower than using the empty porous titanium scaffold. Copyright © 2016 John Wiley & Sons, Ltd.

Functionalization of multilayered DNA-coatings with bone morphogenetic protein 2.

The focus of the present study was to functionalize multilayered DNA-coatings with the osteoinductive factor bone morphogenetic protein 2 (BMP-2) using different loading modalities. The multilayered DNA-coatings were built up from either poly-d-lysine (PDL) or poly(allylamine hydrochloride) (PAH) and DNA using electrostatic self-assembly (ESA). The amounts of BMP-2 loaded into the multilayered DNA-coatings and its subsequent release characteristics were determined using radiolabeled BMP-2. Additionally, the effect of BMP-2 functionalized multilayered DNA-coatings on the in vitro behavior of bone marrow-derived osteoblast-like cells was evaluated in terms of proliferation, differentiation, mineralization, and cell morphology. The results demonstrate the feasibility of multilayered DNA-coatings to be functionalized by embedding BMP-2 according to three different loading modalities: superficial (s), deep (d), and double-layer (dl). BMP-2 was incorporated proportionally into the multilayered DNA-coatings as: s+(4*d)=dl. All differently loaded multilayered DNA-coatings showed an initial burst release followed by an incremental sustained release of the remaining BMP-2. In vitro experiments demonstrated that the loaded factor remained biologically active, as an accelerated calcium deposition was observed on s- and dl-loaded multilayered DNA-coatings, without affecting cell proliferation. In contrast, d-loaded multilayered DNA-coatings influenced osteoblast-like cell behavior by decreasing the deposition of calcium.

Cyto- and histocompatibility of multilayered DNA-coatings on titanium.

DNA-containing biomaterial coatings offer potential beneficial effects for both soft and hard tissue implants because of the structural properties of DNA. In the current study, the aim was to assess the in vitro cyto- and in vivo histocompatibility of multilayered DNA-coatings generated using the electrostatic self-assembly technique, with poly-D-lysine or poly(allylamine hydrochloride) as the cationic counterparts of anionic DNA. Multilayered DNA-coatings were fabricated on titanium substrates. Noncoated titanium substrates served as controls. In vitro experiments with rat primary dermal fibroblasts (RDF) assessing their viability were performed using a Live/Dead assay and an MTT-based assay. The presence of multilayered DNA-coatings did not affect RDF cell viability. On the other hand, an increased proliferation was demonstrated on both types of multilayered DNA-coatings. An in vivo rat model was used to study the soft tissue histocompatibility of subcutaneously inserted implants during implantation periods of 4 and 12 weeks. Light microscopic analysis revealed that all implants were surrounded by a fibrous capsule containing alpha-smooth muscle actin, and that the presence of a multilayered DNA-coating did not induce any adverse effects in terms of inflammation and wound healing. Histomorphometrically, no significant differences in capsule quality or thickness were observed dependent on multilayered DNA-coating or implantation period. The cyto- and histocompatibility of multilayered DNA-coatings demonstrated in this study allows their use and functionalization with appropriate compounds to modulate cell and tissue responses in dental and medical implantology.

Toward accelerated bone regeneration by altering poly(D,L-lactic-co-glycolic) acid porogen content in calcium phosphate cement.

This work aimed to compare in vitro degradation of dense PLGA microspheres and milled PLGA particles as porogens within CPC, considering that the manufacturing of milled PLGA is more cost-effective when compared with PLGA microspheres. Additionally, we aimed to examine the effect of porogen amount within CPC/PLGA on degradation and bone formation. Our in vitro results showed no differences between both forms of PLGA particles (as porogens in CPC; spherical for microspheres, irregular for milled) regarding morphology, porosity, and degradation. Using milled PLGA as porogens within CPC/PLGA, we evaluated the effect of porogen amount on degradation and bone forming capacity in vivo. Titanium landmarks surrounded by CPC/PLGA with 30 and 50 wt % PLGA, were implanted in forty femoral bone defects of twenty male Wistar rats. Histomorphometrical results showed a significant temporal decrease in the amount of CPC, for both formulas, and confirmed that 50 wt % PLGA degrades faster than 30 wt%, and allows for a 1.5-fold higher amount of newly formed bone. Taken together, this study demonstrated that (i) milled PLGA particles perform equal to PLGA microspheres, and (ii) tuning of the PLGA content in CPC/PLGA is a feasible approach to leverage material degradation and bone formation.

Gelation and biocompatibility of injectable alginate-calcium phosphate gels for bone regeneration.

An emerging approach toward development of injectable, self-setting, and fully biodegradable bone substitutes involves the combination of injectable hydrogel matrices with a dispersed phase consisting of nanosized calcium phosphate particles. Here, novel injectable composites for bone regeneration have been developed based on the combination of ultrapure alginate as the matrix phase, crystalline CaP [monetite and poorly crystalline hydroxyapatite (HA)] powders as both a dispersed mineral phase and a source of calcium for cross-linking alginate, glucono-delta-lactone (GDL) as acidifier and glycerol as both plasticizer and temporary sequestrant. The composites were maximized with respect to CaP content to obtain the highest amount of osteoconductive filler. The viscoelastic and physicochemical properties of the precursor compounds and composites were analyzed using rheometry, elemental analysis (for calcium release and uptake), acidity [by measuring pH in simulated body fluid (SBF)], general biocompatibility (subcutaneous implantation in rabbits), and osteocompatibility (implantation in femoral condyle bone defect of rabbits). The gelation of the resulting composites could be controlled from seconds to tens of minutes by varying the solubility of the CaP phase (HA vs. monetite) or amount of GDL. All composites mineralized extensively in SBF for up to 11 days. In vivo, the composites also disintegrated upon implantation in subcutaneous or bone tissue, leaving behind less degradable but osteoconductive CaP particles. Although the composites need to be optimized with respect to the available amount of calcium for cross-linking of alginate, the beneficial bone response as observed in the in vivo studies render these gels promising for minimally invasive applications as bone-filling material.