Reconstruction of critical-size bone defects (CSDs) with complex defect morphologies remains clinically challenging. The desire to avoid autograft harvesting has prompted an increasing quest for adequate synthetic bone grafting materials. The clinical success rates, which have been achieved with bioactive β-tricalcium phosphate granules (TCP-G) demonstrate that these materials have become an excellent alternative graft choice. In order to improve surgical handling properties, TCP-G have been combined with natural polymers for creating paste- and foam-like materials, which can easily be molded into any desired shape when grafting a given bony defect or deploying them with a syringe. This study assessed the effect of a TCP paste (TCP-P) and a TCP-foam (TCP-F) bone grafting material as compared to TCP-G on bone formation and osteogenic marker expression after 1, 3, 6, 12, and 18 months of implantation in CSD in the sheep scapula and tested the hypothesis that the addition of natural polymers would not diminish the osteogenic properties of TCP-P and TCP-F. The bone and bone graft material area fractions were determined histomorphometrically in order to quantify bone formation and bone graft material resorption. Immunohistochemical analysis of collagen type I, osteocalcin, and bone sialoprotein expression in the various cell and matrix components of the bone tissue was performed on resin-embedded sections for characterizing the osteogenic and bioactive properties of the test materials. By 6 months, all three TCP materials facilitated excellent defect regeneration with further bone remodeling at 12 and 18 months. TCP-F and TCP-P induced greater osteocalcin expression and exhibited more advanced graft material resorption at 1 and 6 months, respectively. At 18 months, all three grafting materials were almost fully resorbed with the original bony architecture being restored. Taken together, the hyaluronic acid and methylcellulose components in TCP-P and porcine collagen components in TCP-F did not diminish the osteogenic capacity of TCP-P and TCP-F, which exhibited an even slightly higher resorbability and enhancement effect on OC expression by osteoblasts.

Peri-implant disease due to bacterial infection is the most common cause for dental implant failure. Similarly, surgical site infections (SSI) after orthopedic surgery utilizing Ti-based fracture fixation devices pose a dire clinical problem. Although it is well-known that fullerenes possess excellent biocompatibility, antibacterial and anti-inflammatory properties, their use for surface functionalization of dental and orthopedic implants is poorly investigated. Previously, we established a high-vacuum sublimation process which facilitates coating Ti-6Al-4V implants with complex geometry and a threaded screw design with homogeneous and stable C60 films. This work assessed the effect of fullerene coated zirconia and titanium alloy screw-shaped implants subjected to various surface treatments on the osteogenic, antibacterial and anti-inflammatory properties of these implant surfaces in vitro. Quantitative analysis of osteogenic marker expression by MC3T3-E1 cells was used to characterize the osteogenic properties of the test surfaces. To evaluate the anti-inflammatory properties, expression levels of proinflammatory cytokines from macrophages under LPS challenge were assessed, and the anti-bacterial rate of different surface variants was evaluated by quantification of E.coli adherence and growth. This was in addition to determining the antibacterial rate against P.gingvalis for permucosal surface variants. The results showed that fullerene functionalization rendered excellent antibacterial properties of the implant surfaces and minimized the inflammatory response to zirconia dental implants, while yielding no benefit for osteogenic differentiation. Therefore, a fullerene surface modification of the per-mucosal portion of dental implants or of fracture fixation devices may hold great promise for designing dental implants as well as fracture fixation devices which an enhanced capacity of withstanding the bacterial and inflammatory challenges leading to peri-implantitis and SSI.

Ceramic additive manufacturing currently relies on binders or high-energy lasers, each with limitations affecting final product quality and suitability for medical applications. To address these challenges, our laboratory has devised a surface activation technique for ceramic particles that eliminates the necessity for polymer binders or high-energy lasers in ceramic additive manufacturing. We utilized this method to 3D print bioactive SiC orthopedic screws and evaluated their properties. The study's findings reveal that chemical oxidation of SiC activated its surface, enabling 3D printing of orthopedic screws in a binder jet printer. Post-processing impregnation with NaOH and/or NH4OH strengthened the scaffold by promoting silica crystallization or partial conversion of silicon oxide into silicon nitride. The silica surface of the SiC 3D printed orthopedic screws facilitated osteoblast and neuron adhesion and extensive axon synthesis. The silicate ions released from the 3D printed SiC screws favorably modulated macrophage immune responses toward an M1 phenotype as indicated by the inhibition of TNFα secretions and of reactive oxygen species (ROS) expression along with the promotion of IL6R shedding. In contrast, under the same experimental conditions, Ti ions released from Ti6Al4V discs promoted macrophage TNFα secretion and ROS expression. In vivo tests demonstrated direct bone deposition on the SiC scaffold and a strong interfacial bond between the implanted SiC and bone. Immunostaining showed innervation, mineralization, and vascularization of the newly formed bone at the interface with SiC. Taken altogether, the 3D printed SiC orthopedic screws foster a favorable environment for wound healing and bone regeneration. The novel 3D printing method, based on ceramic surface activation represents a significant advancement in ceramic additive manufacturing and is applicable to a wide variety of materials.

Lately, the potential risk of disease transmission due to the use of bovine-derived bone substitutes has become obvious, demonstrating the urgent need for a synthetic grafting material with comparable bioactive behaviour and properties. Therefore, the effect of a synthetic hydroxyapatite (HA) (Osbone (R)) bone grafting material on bone regeneration was evaluated 2 weeks, 1 month, and 3, 6, 12 and 18 months after implantation in critical-size bone defects in the ovine scapula and compared to that of a bovine-derived HA (Bio-Oss (R)) and beta-tricalcium phosphate (TCP) (Cerasorb (R) M). New bone formation and the biodegradability of the bone substitutes were assessed histomorphometrically. Hard tissue histology and immunohistochemical analysis were employed to characterize collagen type I, alkaline phosphatase, osteocalcin, as well as bone sialoprotein expression in the various cell and matrix components of the bone tissue to evaluate the bioactive properties of the bone grafting materials. No inflammatory tissue response was detected with any of the bone substitute materials studied. After 3 and 6 months, beta-TCP (Cerasorb (R) M) showed superior bone formation when compared to both HA-based materials (3 months: beta-TCP 55.65 +/- 2.03% vs. SHA 49.05 +/- 3.84% and BHA 47.59 +/- 1.97%; p <= 0.03; 6 months: beta-TCP 62.03 +/- 1.58%; SHA: 55.83 +/- 2.59%; BHA: 53.44 +/- 0.78%; p <= 0.04). Further, after 12 and 18 months, a similar degree of bone formation and bone-particle contact was noted for all three bone substitute materials without any significant differences. The synthetic HA supported new bone formation, osteogenic marker expression, matrix mineralization and good bone-bonding behaviour to an equal and even slightly superior degree compared to the bovine-derived HA. As a result, synthetic HA can be regarded as a valuable alternative to the bovine-derived HA without the potential risk of disease transmission.

(1) Background: The desire to avoid autograft harvesting in implant dentistry has prompted an ever-increasing quest for bioceramic bone substitutes, which stimulate osteogenesis while resorbing in a timely fashion. Consequently, a highly bioactive silicon containing calcium alkali orthophosphate (Si-CAP) material was created, which previously was shown to induce greater bone cell maturation and bone neo-formation than β-tricalcium phosphate (β-TCP) in vivo as well as in vitro. Our study tested the hypothesis that the enhanced effect on bone cell function in vitro and in sheep in vivo would lead to more copious bone neoformation in patients following sinus floor augmentation (SFA) employing Si-CAP when compared to β-TCP. (2) Methods: The effects of Si-CAP on osteogenesis and Si-CAP resorbability were evaluated in biopsies harvested from 38 patients six months after SFA in comparison to β-TCP employing undecalcified histology, histomorphometry, and immunohistochemical analysis of osteogenic marker expression. (3) Results: Si-CAP as well as β-TCP supported matrix mineralization and bone formation. Apically furthest away from the original bone tissue, Si-CAP induced significantly higher bone formation, bone-bonding (bone-bioceramic contact), and granule resorption than β-TCP. This was in conjunction with a higher expression of osteogenic markers. (4) Conclusions: Si-CAP induced higher and more advanced bone formation and resorbability than β-TCP, while β-TCP's remarkable osteoconductivity has been widely demonstrated. Hence, Si-CAP constitutes a well-suited bioactive graft choice for SFA in the clinical arena.

Introduction: Recently, efforts towards the development of patient-specific 3D printed scaffolds for bone tissue engineering from bioactive ceramics have continuously intensified. For reconstruction of segmental defects after subtotal mandibulectomy a suitable tissue engineered bioceramic bone graft needs to be endowed with homogenously distributed osteoblasts in order to mimic the advantageous features of vascularized autologous fibula grafts, which represent the standard of care, contain osteogenic cells and are transplanted with the respective blood vessel. Consequently, inducing vascularization early on is pivotal for bone tissue engineering. The current study explored an advanced bone tissue engineering approach combining an advanced 3D printing technique for bioactive resorbable ceramic scaffolds with a perfusion cell culture technique for pre-colonization with mesenchymal stem cells, and with an intrinsic angiogenesis technique for regenerating critical size, segmental discontinuity defects in vivo applying a rat model. To this end, the effect of differing Si-CAOP (silica containing calcium alkali orthophosphate) scaffold microarchitecture arising from 3D powder bed printing (RP) or the Schwarzwalder Somers (SSM) replica fabrication technique on vascularization and bone regeneration was analyzed in vivo. In 80 rats 6-mm segmental discontinuity defects were created in the left femur.Methods: Embryonic mesenchymal stem cells were cultured on RP and SSM scaffolds for 7d under perfusion to create Si-CAOP grafts with terminally differentiated osteoblasts and mineralizing bone matrix. These scaffolds were implanted into the segmental defects in combination with an arteriovenous bundle (AVB). Native scaffolds without cells or AVB served as controls. After 3 and 6 months, femurs were processed for angio-µCT or hard tissue histology, histomorphometric and immunohistochemical analysis of angiogenic and osteogenic marker expression.Results: At 3 and 6 months, defects reconstructed with RP scaffolds, cells and AVB displayed a statistically significant higher bone area fraction, blood vessel volume%, blood vessel surface/volume, blood vessel thickness, density and linear density than defects treated with the other scaffold configurations.Discussion: Taken together, this study demonstrated that the AVB technique is well suited for inducing adequate vascularization of the tissue engineered scaffold graft in segmental defects after 3 and 6 months, and that our tissue engineering approach employing 3D powder bed printed scaffolds facilitated segmental defect repair.

Sinus floor augmentation (SFA) has become a well-recognized procedure for site development in the atrophic posterior maxilla prior to dental implant placement. There has been an increasing search for relatively rapidly resorbable bone substitutes that enhance bone formation and facilitate bone regeneration with complete replacement by functional bone tissue. This has led to the development of a glassy crystalline silica-containing calcium alkali orthophosphate (Si-CAOP). The specific aim of this study was to assess histologically the effect of this grafting material on osteogenesis and osteogenic marker expression and to compare this effect to that of β-tricalcium phosphate (β-TCP) in human biopsies obtained 6 months after SFA. Cylindrical biopsies, harvested 6 months after SFA, were processed for immunohistochemical analysis of sawed hard tissue sections using primary antibodies specific to osteocalcin (OC), collagen type I (Col I), bone sialoprotein (BSP), and alkaline phosphatase (ALP). Furthermore, the bone and particle area fraction were measured histomorphometrically in all biopsies in order to characterize bone formation and degradation of the bone grafting materials. Both biomaterials facilitated osteoblast differentiation and bone regeneration of resorbed alveolar ridges, resulting in sufficient bone formation for supporting dental implants. Si-CAOP specimens displayed more advanced bone formation and significantly greater particle degradation than TCP sites. This was accompanied by significantly higher expression of OC, Col I, BSP, and ALP in the osteoid in contact with the degrading particles. These promising results indicate that the Si-CAOP material has great potential use for SFA in humans, thereby confirming its superiority to β-TCP. A prospective study involving a larger patient number, a split-mouth design and analysis of angiogenic properties and of cone-beam CT-data for assessing the volume stability of the augmented area is warranted to further confirm the high osteogenic capacity of Si-CAOP for orofacial bone regeneration.

Surgical site infections after orthopaedic surgery using fracture fixation devices or endosseous implants create major surgical challenges with severe adverse effects, such as osteomyelitis. These infections are frequently caused by Staphylococcus aureus, often with high resistance to antibiotics, such as methicillin-resistant Staphylococcus aureus (MRSA). Due to the formation of impenetrable biofilms on implant surfaces, systemic antibiotic treatment has become exceedingly difficult. New solutions are pursued by combining several drugs using a controlled delivery system from specifically engineered implant surfaces. A sol-gel coating on titanium implants was previously developed with 20 wt % vancomycin and 30 wt % farnesol, with suppression of MRSA in vitro. The present study investigated the efficacy of sol-gel film coatings for controlled dual local delivery over 4 weeks utilising a rat infection model. The findings confirmed the viability of this new concept in vivo based on the differences observed between coatings containing vancomycin alone (SGV) and the dual-drug-containing coating with vancomycin and farnesol (SGVF). While both the SGVF and SGV coatings facilitated excellent preservation of the osseous microarchitecture, SGVF coating displayed a slightly higher potency for suppressing MRSA infiltration than SGV, in combination with a lower reactive bone remodelling activity, most likely by disturbing biofilm formation. The next step for advancing the concept of dual-drug delivery from sol-gel coatings to the clinic and confirming the promising effect of the SGVF coatings on reactive bone remodelling and suppressing MRSA infiltration is a study in a larger animal species with longer time points.

Computed tomography (CT) with hard X-rays is a mature technique that is in regular use to depict the interior of opaque specimens with spatial resolutions up to the micrometre range (microtomography or µCT). Short acquisition times and sophisticated contrast modes are accessible when synchrotron light sources are combined with microtomography—SR-µCT. Both features render SR-µCT as excellent probe to study delicate samples in situ, for example under mechanical load by deploying corresponding sample environments. The so-called TomoPress is such a device available within the public user programme of tomography beamline ID19 of the European Synchrotron Radiation Facility (ESRF). It allows one to study samples under high axial load (up to 500 N) with high spatial resolution up to the micrometer range. Different gauges are installed to allow online monitoring of the applied force. Constant humidity, temperature and wetting are routinely available as well. The article shall outline basic design principles of the press as well as parameters for its utilisation in a descriptive manner. Selected examples underline the potential of the device for such diverse fields as biomedical research, life sciences and materials research.

Compared to the currently clinically available bone grafting materials for alveolar ridge augmentation, there is a great demand for bioactive bone substitutes with higher resorbability, which enhance osteogenesis at the same time. This has prompted the development of a silicon-doped rapidly resorbable calcium alkali orthophosphate (Si-CAOP) and silicon-doped β-tricalcium phosphate (Si-TCP). This study evaluated the effect of these two particulate graft materials as compared to the currently clinically used β-TCP on bone formation and osteogenic marker expression after 2 weeks, 1, 3, 6, 12, and 18 months of implantation in critical size defects in the sheep scapula. Immunohistochemical analysis of collagen type I, alkaline phosphatase, and osteocalcin expression was performed on resin embedded sections. The bone and particle area fraction and the bone-biomaterial contact were determined histomorphometrically. After 2 weeks and 1 month defects grafted with Si-CAOP displayed a significantly greater bone area fraction, bone-particle-contact, osteogenic marker expression and significantly lower particle area fraction than defects grafted with Si-TCP and TCP. By 3 and 6 months all materials studied mediated excellent defect regeneration with further bone remodeling at 12 and 18 months. Taken together, Si-CAOP induced the most expeditious bone regeneration of critical size defects in the sheep scapula. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2018. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 594-614, 2019.

Gellan gum hydrogels functionalized with alkaline phosphatase were enzymatically mineralized with phosphates in mineralization medium containing calcium (Ca) and zinc (Zn) to improve their suitability as biomaterials for bone regeneration. The aims of the study were to endow mineralized hydrogels with antibacterial activity by incorporation of Zn in the inorganic phase, and to investigate the effect of Zn incorporation on the amount and type of mineral formed, the compressive modulus of the mineralized hydrogels and on their ability to support adhesion and growth of MC3T3-E1 osteoblast-like cells. Mineralization medium contained glycerophosphate (0.05 m) and three different molar Ca:Zn ratios, 0.05:0, 0.04:0.01 and 0.025:0.025 (all mol/dm3 ), hereafter referred to as A, B and C, respectively. FTIR, SAED and TEM analysis revealed that incubation for 14 days caused the formation of predominantly amorphous mineral phases in sample groups A, B and C. The presence of Zn in sample groups B and C was associated with a drop in the amount of mineral formed and a smaller mineral deposit morphology, as observed by SEM. ICP-OES revealed that Zn was preferentially incorporated into mineral compared to Ca. Mechanical testing revealed a decrease in compressive modulus in sample group C. Sample groups B and C, but not A, showed antibacterial activity against biofilm-forming, methicillin-resistant Staphylococcus aureus. All sample groups supported cell growth. Zn incorporation increased the viable cell number. The highest values were seen on sample group C. In conclusion, the sample group containing the most Zn, i.e. group C, appears to be the most promising. Copyright © 2015 John Wiley & Sons, Ltd.

Little is known regarding the associations between sex-hormone levels, sex, body mass index (BMI), age, other host factors and biomaterial stimulated bone regeneration in the human craniofacial skeleton. The aim of this study was to elucidate the associations between these factors and bone formation after sinus floor augmentation procedures (SFA) utilizing a bioactive tricalcium phosphate (TCP) bone grafting material. We conducted a prospective study in a human population in which 60 male and 60 female participants underwent SFA and dental implant placement using a staged approach. BMI as well as levels of serum estradiol (E2), total testosterone (TT), and the free androgen index (FAI) were measured by radioimmunoassay and electrochemoluminescent-immunoassay. At implant placement, 6 months after SFA, bone biopsy specimens were harvested for hard tissue histology, the amount of bone formation was evaluated by histomorphometry and immunohistochemical analysis of osteogenic marker expression. The Wilcoxon rank-sum U test, Spearman correlations and linear regression analysis were used to explore the association between bone formation and BMI, hormonal and other host factors. BMI and log E2 were significantly positively associated with bone formation in male individuals (p < 0.05). Histomorphometry revealed trends toward greater bone formation and osteogenic marker expression with non-smokers compared to smokers. In male patients, higher E2 levels and higher BMI enhanced TCP stimulated craniofacial i.e. intramembranous bone repair.

Due to the significant increase in dental implants placements and in alveolar ridge augmentation procedures over the last two decades, there has been an ever increasing demand for adequate bone grafting materials. Consequently, numerous bone grafting materials have been investigated. This includes calcium phosphate-based grafting materials such as synthetic as well as corallline or bovine-derived hydroxyapatites, tricalcium phosphate ceramics, biphasic hydroxyapatite tricalcium phosphates, calcium carbonates, bioactive glasses and glass ceramics, and demineralized freeze-dried bone allografts. More recently injectable and mouldable cements, putty like materials as well as scaffolds for craniofacial bone tissue engineering have been developed. Furthermore the combination of grafting materials with growth factors has been explored. However, compared to the bone substitute materials which are currently clinically available, there is a significant need for bone substitute materials which degrade more rapidly, but still stimulate osteogenesis at the same time. This has initiated an ever increasing search for bioactive rapidly resorbable bone grafting materials, which exhibit good bone bonding behavior by stimulating enhanced bone formation at the interface in combination with a high degradation rate. A silica containing calcium alkali othophophate material, which meets these requirements, has been shown to render excellent bone regeneration results in the maxillary sinus floor 6 months after sinus floor augmentation in humans in a first clinical study. Furthermore, current research efforts include the optimization of resorbable bone grafting cements as well as of scaffolds and concepts for various tissue engineering approaches.

This study examines the effect of a hyaluronic acid (HyAc) containing tricalcium phosphate putty scaffold material (TCP-P) and of a particulate tricalcium phosphate (TCP-G) graft on bone formation, volume stability and osteogenic marker expression in biopsies sampled 6 months after bilateral sinus floor augmentation (SFA) in 7 patients applying a split-mouth design. 10% autogenous bone chips were added to the grafting material during surgery. The grain size of the TCP granules was 700 to 1400 µm for TCP-G and 125 to 250 µm and 500 to 700 µm (ratio 1:1) for TCP-P. Biopsies were processed for immunohistochemical analysis of resin-embedded sections. Sections were stained for collagen type I (Col I), alkaline phosphatase (ALP), osteocalcin (OC) and bone sialoprotein (BSP). Furthermore, the bone area and biomaterial area fraction were determined histomorphometrically. Cone-beam CT data recorded after SFA and 6 months later were used for calculating the graft volume at these two time points. TCP-P displayed more advantageous surgical handling properties and a significantly greater bone area fraction and smaller biomaterial area fraction. This was accompanied by significantly greater expression of Col I and BSP and in osteoblasts and osteoid and a less pronounced reduction in grafting volume with TCP-P. SFA using both types of materials resulted in formation of sufficient bone volume for facilitating stable dental implant placement with all dental implants having been in function without any complications for 6 years. Since TCP-P displayed superior surgical handling properties and greater bone formation than TCP-G, without the HyAc hydrogel matrix having any adverse effect on bone formation or graft volume stability, TCP-P can be regarded as excellent grafting material for SFA in a clinical setting. The greater bone formation observed with TCP-P may be related to the difference in grain size of the TCP granules and/or the addition of the HyAc.

Bioactive ceramics are known to bond directly to bone and to stimulate bone formation. This has led to their constantly increasing clinical use. Numerous studies have contributed to enhancing our understanding regarding the stimulatory effect of bioactive calcium phosphates on osteoblast function and bone tissue formation. Significant progress has been made in revealing solution-mediated surface reactions that take place close to the surface of bioactive ceramics. Advanced surface analysis methods have been combined with molecular techniques in order to facilitate a better understanding of the surface transformations of bioactive, resorbable ceramics and glasses and the serum protein adsorption events associated with the immersion in biological fluids. Increasing knowledge has been generated regarding the effect of bioactive ceramics on osteoblast differentiation in vitro. Furthermore, the effect of various bioactive ceramics on osteoblast differentiation and tissue maturation in vivo has been studied, thereby allowing correlation of in vitro and in vivo events. More recent research efforts provided insight into integrin-mediated cell adhesion and the subsequently activated intracellular signaling pathways that regulate cell function including differentiation and cell survival. Once reaction pathways are clearly identified, materials can be created that preferentially boost the osteogenic cascade and thus are optimally tailored toward their clinical application.

Computed tomography, frequently called CT, with high spatial resolution up to the (sub-)micrometer range (μCT) can be considered as microscopy in three dimensions. The spatial resolution and contrast in μCT volume images is significantly increased when synchrotron light sources are applied instead of laboratory-based X-ray tubes. This fact is related to the orders of magnitude higher photon flux density available, the nearly parallel beam propagation as well as the spatial coherence of the beam at the sample position. In this chapter, synchrotron-based microtomography using hard X-rays is employed to study bioceramic-supported bone regeneration. The high contrast and excellent signal-to-noise ratio in the tomographic images are the basis to segment the data sets. Thus, volume renderings are acquired which can be considered as an extension of classical histology into three dimensions but in a truly nondestructive manner. The principles of the techniques for acquiring as well as analyzing the images are introduced and described in this chapter including a detailed review on the available literature in order to facilitate further studies by the reader. To demonstrate the potential of synchrotron-based microtomography in biomaterials research, the example of regenerating human bone tissue with the support of clinically established bone substitute materials is chosen. Furthermore, results from an animal study are shown in which various resorbable ceramics were compared for bone grafting.

Early establishment of angiogenesis is critical for bone tissue engineering. Recently, a technique was introduced, which is based on the idea of using axial vascularization of the host tissues in engineered grafts, namely the “intrinsic angiogenesis chamber” technique, which utilizes an artery and a vein to construct an AV-Bundle. The aim of this study was to evaluate the effect of varying scaffold architecture of calcium alkali orthophosphate scaffolds (CAOP), resulting from two different fabrication procedures, namely 3D printing (RP) or a Schwarzwalder-Somers replica technique (SSM), on angiogenesis in vivo when combining a microvascular technique with bioceramic scaffolds colonized with stem cells for bone tissue engineering. 32 adult female Wistar rats, in which critical size segmental discontinuity defects 6 mm in length were created in the left femur, were divided into 4 groups, group 1 received a RP scaffold colonized with rat stem cells after 7d of dynamic cell culture and an AV-Bundle (AVB), group 2 a SSM scaffold with rat stem cells after 7d of dynamic cell culture and an AVB, group 3 a RP control scaffold (without cells and AVB), group 4 a SSM control scaffold (without cells and AVB). After 3 and 6 months, angiomicro-CT after perfusion with a contrast agent, image reconstruction, histomorphometric and immunohistochemical analysis utilizing antibodies to collagen IV, vWF and CD-31 were performed. At 6 months, a statistically significant higher blood vessel volume%, blood vessel surface/volume, blood vessel thickness, blood vessel density and blood vessel linear density was observed with RP scaffolds with cells and AVB than with the other groups. At 6 mths, RP with cells and AVB displayed the highest expression of collagen IV (score 2.75), CD31 (score 2.75) and vWF (score 2.6), which is indicative of highly dense blood vessels. Both angio-CT and immunohistochemical analysis demonstrated that AVB is an efficient technique for achieving scaffold vascularization in critical size segmental defects after 3 and 6 months of implantation.

Sinus floor augmentation (SFA) has become a well-established pre-implantology procedure for alveolar ridge augmentation of the posterior maxilla. Using bioceramic bone substitutes avoids second-site surgery for autograft harvesting. Compared to the bone substitutes which are currently clinically available, there is a significant need for bone substitutes which degrade more rapidly, but still stimulate osteogenesis at the same time. This has led to the development of bioactive, rapidly resorbable calcium alkali orthophosphate (CAOP) materials, which have a greater solubility than tricalcium phosphate. In this study the biodegradability and effect of a silica containing CAOP (Si-CAOP) on osteogenesis was evaluated in human biopsies sampled 6 months after SFA and compared to that of TCP utilizing hard tissue histology, histomorphometry and immunohistochemical analysis of osteogenic marker expression. Both materials facilitated bone formation and matrix mineralization, which were still actively progressing from the sinus floor in an apical direction 6 months after SFA. With the Si-CAOP grafting material however, bone formation, the bone-biomaterial-contact, i.e. bone-bonding, and particle degradation were significantly greater compared to TCP in the apical region of the biopsies, i.e. at the largest distance from the native bone of the sinus floor. This was accompanied by greater expression of Col I, BSP and OC in the newly formed bone tissue in the Si-CAP samples compared to TCP. Six months after implantation Si-CAOP facilitated greater bone formation and biodegradability than the TCP graft material, whose excellent osteoconductive properties have been widely documented. Consequently, Si-CAOP can be regarded as excellent grafting material for SFA in a clinical setting.