Bifunctional bone substitute materials for bone defect treatment after bone tumor resection.

Aggressive benign, malignant and metastatic bone tumors can greatly decrease the quality of patients' lives and even lead to substantial mortality. Several clinical therapeutic strategies have been developed to treat bone tumors, including preoperative chemotherapy, surgical resection of the tumor tissue, and subsequent systemic chemo- or radiotherapy. However, those strategies are associated with inevitable drawbacks, such as severe side effects, substantial local tumor recurrence, and difficult-to-treat bone defects after tumor resection. To overcome these shortcomings and achieve satisfactory clinical outcomes, advanced bifunctional biomaterials which simultaneously promote bone regeneration and combat bone tumor growth are increasingly advocated. These bifunctional bone substitute materials fill bone defects following bone tumor resection and subsequently exert local anticancer effects. Here we describe various types of the most prevalent bone tumors and provide an overview of common treatment options. Subsequently, we review current progress regarding the development of bifunctional bone substitute materials combining osteogenic and anticancer efficacy. To this end, we categorize these biomaterials based on their anticancer mechanism deriving from i) intrinsic biomaterial properties, ii) local drug release of anticancer agents, and iii) oxidative stress-inducing and iv) hyperthermia-inducing biomaterials. Consequently, this review offers researchers, surgeons and oncologists an up-to-date overview of our current knowledge on bone tumors, their treatment options, and design of advanced bifunctional biomaterials with strong potential for clinical application in oncological orthopedics.

Nanoindentation and nano-scratching of hydroxyapatite coatings for resorbable magnesium alloy bone implant applications.

The corrosion rate of Mg alloys is currently too high for viable resorbable implant applications. One possible solution is to coat the alloy with a hydroxyapatite (HA) layer to slow the corrosion and promote bone growth. As such coatings can be under severe stresses during implant insertion, we present a nano-mechanical and nano-tribological investigation of RF-sputtered HA films on AZ31 Mg alloy substrates. EDX and XRD analysis indicate that as-deposited coatings are amorphous and Ca-deficient whereas rapid thermal annealing results in c-axis orientation and near-stoichiometric composition. Analysis of the nanoindentation data using a thin film model shows that annealing increases the coating's intrinsic hardness (H) and strain at break (H/E) values, from 2.7 GPa to 9.4 GPa and from 0.043 to 0.079, respectively. In addition, despite being rougher, the annealed samples display better wear resistance; a sign that the rapid thermal annealing does not compromise their interfacial strength and that these systems have potential for resorbable bone implant applications.

Species-independent stimulation of osteogenic differentiation induced by osteoclasts.

The coupling of bone resorption and bone formation is well-recognized in the bone remodeling process, in which osteoblasts and osteoclasts are key players. However, the anabolic effect of human primary osteoclasts has rarely been reported as mouse and cell line derived osteoclasts were mostly used in previous reports. Therefore, a comprehensive comparison of mouse and human osteoclasts and their corresponding functions is needed to study cell-cell interactions between osteoclasts and osteoblasts. Osteoclasts from mouse and human origin were generated, characterized and compared, after which their anabolic effects on the osteogenic differentiation of mouse and human MSCs were assessed. Both murine RAW264.7 derived osteoclasts (mOCs) and primary human osteoclasts (hOCs) derived from buffy coats characteristically displayed multinuclearity, marked integrin ß3 expression and enhanced TRAP activity. Despite comparable cell size, mOCs showed higher osteoclast density (number of osteoclasts per cm2 culture dish) and osteoclast nuclearity (average number of nuclei per osteoclast), but lower TRAP activity compared to hOCs. Culturing primary rat and human bone marrow MSCs with the conditioned medium of mOCs or hOCs showed anabolic effects regarding the osteogenic differentiation of MSCs with superiority of hOCs over mOCs. We conclude that despite morphological and functional differences between mouse and human osteoclasts, their secretory factors evoke similar anabolic effects on MSC osteogenic differentiation.

Histological evaluation of titanium fiber mesh-coated implants in a rabbit femoral condyle model.

OBJECTIVES: This study was aimed to comparatively evaluate new bone formation into the pores of a flexible titanium fiber mesh (TFM) applied on the surface of implant. METHODS: Twenty-eight custom made cylindrical titanium implants (4 ×10 mm) with and without a layer of two different types of TFM (fiber diameter of 22 µm and 50 µm, volumetric porosity ~70%) were manufactured and installed bilaterally in the femoral condyles of 14 rabbits. The elastic modulus for these two TFM types was ~20 GPa and ~5 GPa respectively, whereas the solid titanium was ~110 GPa. The implants (Control, TFM-22, TFM-50) were retrieved after 14 weeks of healing and prepared for histological assessment. The percentage of the bone area (BA%), the bone-to-implant contact (BIC%) and amount were determined. RESULTS: Newly formed bone into mesh porosity was observed for all three types of implants. Histomorphometric analyses revealed significantly higher (~2.5 fold) BA% values for TFM-22 implants (30.9 ± 9.5%) compared to Control implants (12.7 ± 6.0%), whereas BA% for TMF-50 did not significantly differ compared with Control implants. Furthermore, both TFM-22 and TFM-50 implants showed significantly higher BIC% values (64.9 ± 14.0%, ~2.5 fold; 47.1 ± 14.1%, ~2 fold) compared to Control (23.6 ± 17.4%). Finally, TFM-22 implants showed more and thicker trabeculae in the peri-implant region. SIGNIFICANCE: This in vivo study demonstrated that implants with a flexible coating of TFM improve bone formation within the inter-fiber space and the peri-implant region.

Innovative implant design for continuous implant stability: A mechanical and histological experimental study in the iliac crest of goats.

OBJECTIVES: this in vivo study reports on mechanical torque data as well as the biological evaluation up to 6 weeks after placement of implants with a unique wide knife thread design in a goat iliac crest model. We hypothesized that implants with this thread design would show substantial primary stability at a continuous level toward secondary stability. METHODS: 64 MegaGen Anyridge® implants were used with diameters 3.5 mm, 4.0 mm, 5.0 mm and 6.0 mm (n = 8). Implants were placed monocortically in the iliac crest of 16 healthy female Saanen goats, both on the right (for torque measurements) and left side (for histology/-morphometry). Torque-in at implant installation and torque-out at 2 and 6 weeks of implantation was measured, as well as bone-to-implant contact (BIC) and bone-area between the screw threads (BA). RESULTS: Histology showed intimate bone-to-implant contact with a maturating trabecular structure between 2 and 6 weeks. Torque values showed a dependency on implant diameter. For all implant diameters, torque-in values were similar to torque-out values at 2 weeks. At 6 weeks however, all torque-out values were significantly increased. BIC and BA percentages showed similar values for all diameters at both 2 and 6 weeks. CONCLUSIONS: These results prove the absence of a lag-phase in implant stability for MegaGen Anyridge® implants in the goat iliac crest model. The increased torque-out values at 6 weeks without increasing BIC and BA percentages correlate with the observed maturation of bone-to-implant contact quality over time. CLINICAL SIGNIFICANCE: It is a challenge to optimize implants with continuous primary stability and rapid transition into secondary stability to minimize the duration of the lag-phase. The results of this study prove the absence of a lag-phase in implant stability for MegaGen Anyridge® implants. Consequently, the data from this work are important for the treatment of individual patients 'translating' these findings into clinical implant procedures.

Stabilizing dental implants with a fiber-reinforced calcium phosphate cement: An in vitro and in vivo study.

Stabilization of dental implants by means of biomaterials such as bioceramic granules and cements is currently compromised by the poor mechanical properties of these bioceramics. Recently, our group developed a calcium phosphate cement reinforced with poly(vinyl alcohol) fibers with improved flexural strength and toughness. Herein we evaluated the capacity of these fiber-reinforced calcium phosphate cements to stabilize dental implants in vitro and in vivo using a range of mechanical and biological test methods. In vitro, filling of circumferential crestal peri­implant bone defects with synthetic bone analogues with fiber-reinforced calcium phosphate cement demonstrated superior implant stability as compared to fiber-free calcium phosphate cement over a 12-week period. Similarly, filling of circumferential crestal peri­implant bone defects with fiber-reinforced calcium phosphate cement effectively stabilized dental implants installed in a rabbit femoral condyle defect as assessed via both Implant Stability Quotient (ISQ) and torque-out measurements. Moreover, histological and histomorphometric evaluation demonstrated the osteocompatibility of fiber-reinforced calcium phosphate cement, as evidenced by absence of soft tissue ingrowth, direct contact between the bone and cement, and gradual degradation of the biomaterial and replacement by newly-formed bone. These data demonstrate that fiber-reinforced calcium phosphate cement stabilize dental implants during osseointegration. STATEMENT OF SIGNIFICANCE: Dental implants can be placed immediately after a tooth is removed. However, in some cases the implant might not have enough bone surrounding it and becomes loose. To solve this, bioceramics have been used to fill the implant-bone gap. However, these materials have poor mechanical properties and are often not capable to stabilize the implant. Recently, our research group developed a new bone cement that is reinforced with fibers and has, therefore, enhanced mechanical properties. In this study, we have proven that by molding this cement into the implant-bone gap, we stabilize the implant and allow for a direct connection between the implant and the surrounding bone. Using this innovative cement is therefore a safe and efficient way of stabilizing dental implants.

Mechanical aspects of dental implants and osseointegration: A narrative review.

With the need of rapid healing and long-term stability of dental implants, the existing Ti-based implant materials do not meet completely the current expectation of patients. Low elastic modulus Ti-alloys have shown superior biocompatibility and can achieve comparable or even faster bone formation in vivo at the interface of bone and the implant. Porous structured Ti alloys have shown to allow rapid bone ingrowth through their open structure and to achieve anchorage with bone tissue by increasing the bone-implant interface area. In addition to the mechanical properties of implant materials, the design of the implant body can be used to optimize load transfer and affect the ultimate results of osseointegration. The aim of this narrative review is to define the mechanical properties of dental implants, summarize the relationship between implant stability and osseointegration, discuss the effect of metallic implant mechanical properties (e.g. stiffness and porosity) on the bone response based on existing in vitro and in vivo information, and analyze load transfer through mechanical properties of the implant body. This narrative review concluded that although several studies have presented the advantages of low elastic modulus or high porosity alloys and their effect on osseointegration, further in vivo studies, especially long-term observational studies are needed to justify these novel materials as a replacement for current Ti-based implant materials.

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.

Monitoring local delivery of vancomycin from gelatin nanospheres in zebrafish larvae.

BACKGROUND: Infections such as biomaterial-associated infection and osteomyelitis are often associated with intracellular survival of bacteria (eg, Staphylococcus aureus). Treatment of these infections remains a major challenge due to the low intracellular efficacy of many antibiotics. Therefore, local delivery systems are urgently required to improve the therapeutic efficacy of antibiotics by enabling their intracellular delivery. PURPOSE: To assess the potential of gelatin nanospheres as carriers for local delivery of vancomycin into macrophages of zebrafish larvae in vivo and into THP-1-derived macrophages in vitro using fluorescence microscopy. MATERIALS AND METHODS: Fluorescently labeled gelatin nanospheres were prepared and injected into transgenic zebrafish larvae with fluorescent macrophages. Both the biodistribution of gelatin nanospheres in zebrafish larvae and the co-localization of vancomycin-loaded gelatin nanospheres with zebrafish macrophages in vivo and uptake by THP-1-derived macrophages in vitro were studied. In addition, the effect of treatment with vancomycin-loaded gelatin nanospheres on survival of S. aureus-infected zebrafish larvae was investigated. RESULTS: Internalization of vancomycin-loaded gelatin nanospheres by macrophages was observed qualitatively both in vivo and in vitro. Systemically delivered vancomycin, on the other hand, was hardly internalized by macrophages without the use of gelatin nanospheres. Treatment with a single dose of vancomycin-loaded gelatin nanospheres delayed the mortality of S. aureus-infected zebrafish larvae, indicating the improved therapeutic efficacy of vancomycin against (intracellular) S. aureus infection in vivo. CONCLUSION: The present study demonstrates that gelatin nanospheres can be used to facilitate local and intracellular delivery of vancomycin.

Putty-like bone fillers based on CaP ceramics or Biosilicate® combined with carboxymethylcellulose: Characterization, optimization, and evaluation.

Calcium phosphates and bioactive glass ceramics have been considered promising biomaterials for use in surgeries. However, their moldability should be further enhanced. We here thereby report the handling, physicochemical features, and morphological characteristics of formulations consisting of carboxymethylcellulose-glycerol and hydroxyapatite-tricalcium phosphate or Biosilicate® particles. We hypothesized that combining either material with carboxymethylcellulose-glycerol would improve handling properties, retaining their bioactivity. In addition to scanning electron microscopy, cohesion, mineralization, pH, and viscoelastic properties of the novel formulations, cell culture experiments were performed to evaluate the cytotoxicity and cell proliferation. Putty-like formulations were obtained with improved cohesion and moldability. Remarkably, mineralization in simulated body fluid of hydroxyapatite-tricalcium phosphate/carboxymethylcellulose-glycerol formulations was enhanced compared to pure hydroxyapatite-tricalcium phosphate. Cell experiments showed that all formulations were noncytotoxic and that HA-TCP60 and BGC50 extracts led to an increased cell proliferation. We conclude that combining carboxymethylcellulose-glycerol with either hydroxyapatite-tricalcium phosphate or Biosilicate® allows for the generation of moldable putties, improves handling properties, and retains the ceramic bioactivity.