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Abstract The objective was to evaluate the effect of oral administration of strontium (Sr) on the regeneration of a critical bone defect. Twenty Wistar rats were divided into two groups, assessed at 15 and 60 days postoperatively: GSr - oral administration of Sr; and CG - without oral administration of Sr. At 15 and 60 days, blood was collected to measure Sr and calcium (Ca), and the calvaria was removed for histomorphological and histomorphometric analyses. Sr+2 plasmatic concentrations were higher in GSr than in CG. The dosage of Ca+2 showed a small increase in the CG about the GSr. In all evaluated groups, new bone formation was restricted to the edges of the defect, and the residual area was filled with fibrous connective tissue. The oral administration of Sr associated with HA microspheres substituted by the metal, in a concentration of 23 mol%, did not affect the formation of the osteoid matrix.

ABSTRACT Purpose: to evaluate biocompatibility and osteogenic potential of hydroxyapatite/alginate composite after its implantation on rat calvarian critical bone defect. Methods: thirty adults male Wistar rats were randomly distributed into two groups: GHA – critical bone defect filled with hydroxyapatite/alginate composite granules (HA/Alg) and CG – critical bone defect without biomaterial; evaluated at biological points of 15, 45 and 120 days. Results: the histomorphometrically analyses for GHA showed osteoid matrix deposition (OM) among the granules and towards the center of the defect in centripetal direction throughout the study, with evident new bone formation at 120 days, resulting in filling 4/5 of the initial bone defect. For CG, this finding was restricted to the edges of the bone margins and formation of connective tissue on the residual area was found in all biological points. Inflammatory response on GHA was chronic granulomatous type, discrete and regressive for all biological points. Throughout the study, the CG presented mononuclear inflammatory infiltrate diffuse and regressive. Histomorphometry analyses showed that OM percentage was evident for GHA group when compared to CG group in all analyzed periods (p > 0.05). Conclusions: the biomaterial evaluated at this study showed to be biocompatible, bioactive, osteoconductive and biodegradable synchronously with bone formation.

The strategies used to associate peptide arginylglycylaspartic acid (RGD) with calcium phosphate grafts to enhance cell–biomaterial interactions have been controversial in the literature. Several works have demonstrated that RGD-functionalized hydroxyapatite (HA) surfaces improve cell adhesion, whereas others claim that RGD-loaded HA has an inhibitory effect when serum is present in the biological medium. To investigate such contradictory results, we associated RGD with the HA surface using physical adsorption and chemical bonding methods and evaluated the cell adhesion and spreading in pre-osteoblasts culture with and without fetal bovine serum (FBS). The effect of functionalization methods on the physicochemical characteristics of both surfaces was analyzed using multiscale techniques. Adsorption assays of serum allowed us to estimate the impact of the association method on the HA surface’s reactivity. Physically adsorbed RGD did not increase the number of adhered cells due to the weak interactions between the peptide and the surface. Although chemical binding stabilizes RGD on the HA, the functionalization procedure covered the surface with molecules such as (3-aminopropyl)triethoxysilane (APTEs) and carbodiimide, changing the surface’s chemical activity. Serum protein adsorption decreased by 90%, revealing a significant reduction in the surface interactions with molecules of the biological medium. The present study’s findings showed that the RGD’s physical association with HA did not improve cell adhesion and that this phenomenon is highly dependent on the presence of serum proteins.

Bone critical-size defects and non-union fractures have no intrinsic capacity for self-healing. In this context, the emergence of bone engineering has allowed the development of functional alternatives. The aim of this study was to evaluate the capacity of ASC spheroids in bone regeneration using a synergic strategy with 3D-printed scaffolds made from poly (lactic acid) (PLA) and nanostructured hydroxyapatite doped with carbonate ions (CHA) in a rat model of cranial critical-size defect. In summary, a set of results suggests that ASC spheroidal constructs promoted bone regeneration. In vitro results showed that ASC spheroids were able to spread and interact with the 3D-printed scaffold, synthesizing crucial growth factors and cytokines for bone regeneration, such as VEGF. Histological results after 3 and 6 months of implantation showed the formation of new bone tissue in the PLA/CHA scaffolds that were seeded with ASC spheroids. In conclusion, the presence of ASC spheroids in the PLA/CHA 3D-printed scaffolds seems to successfully promote bone formation, which can be crucial for a significant clinical improvement in critical bone defect regeneration.

Electrospinning is one of the most promising fabrication techniques of nanostructured membranes for guided bone regeneration. In association with antibiotics, membranes consisting of fibers with sub-micrometric and nanometric dimensions have been tested to prevent and treat bone infections. Electrospinning was recently applied to produce metallic implant coatings for biofilm inhibition. Despite the numerous in vitro and in vivo studies conducted with nanofibers from several polymeric matrices and fabrication methods, there is no consensus on the best conditions to optimize their antimicrobial activity. This study analyzed recent advances in nanofiber fabrication by the electrospinning technique for clinical applications in the treatment of bone infections. An integrative review from MEDLINE/PubMed, Web of Science, and Scielo databases selected 16 works focused on nanofibers' in vitro and in vivo evaluations. It was found that manufacturing methods significantly influence fiber composition, structure, morphology, pore size, and biodegradability. Thus, standardizing these parameters in the production of nanofibers at an industrial scale is one of the challenges in improving drug loading control on the fiber network and its release profiles. Further in vivo studies need to be conducted to optimize the dose effect of antibiotic-loaded membranes in inhibiting the proliferation of pathogens and inflammatory processes without promoting toxicity and reducing bone regenerating capacity.

Magnetic nanoparticles (MNPs) have been widely used for their potential applications, mainly for the diagnosis and/or therapy (theranostic) of several diseases in the field of nanomedicine, as passive contrast agents, through the opsonization process, or active contrast agents, after their functionalization and the subsequent capture of the signal using various techniques such as magnetic resonance imaging (MRI), optical imaging, nuclear imaging, and ultrasound [...]

A global need exists for new and more effective contrast agents for computed tomography and traditional X-ray modalities. Among the few options available nowadays, limitations imposed by industrial production, performance, and efficacy restrict the use and reduce the potential of both imaging techniques. The use of nanomaterials as new contrast agents for X-ray and computed tomography is an innovative and viable way to increase the options and enhance performance. In this study, we evaluated eight nanomaterials: hydroxyapatite doped with zinc (Zn-HA 10%); hydroxyapatite doped with strontium (Sr-HA 10%); hydroxyapatite without thermal treatment (HA 282 STT); thermally treated hydroxyapatite (HA 212 500 °C and HA 01.256 CTT 1000 °C); hydroxyapatite microspheres (HA microspheres); gold nanoparticles (AuNP); and graphene oxide doped with copper (Cu-GO). The results showed that for both imaging modalities; HA microspheres were the best option, followed by hydroxyapatite thermally treated at 1000 °C. The nanomaterials with the worst results were hydroxyapatite doped with zinc (Zn-HA 10%), and hydroxyapatite doped with strontium (Sr-HA 10%). Our data demonstrated the potential of using nanomaterials, especially HA microspheres, and hydroxyapatite with thermal treatment (HA 01.256 CTT 1000 °C) as contrast agents for X-ray and computed tomography.

Medication-related osteonecrosis of the jaw (MRONJ) is considered as a severe adverse side effect of specific drugs such as anti-resorptive and anti-angiogenic medications. Evidence suggests that MRONJ is linked to invasive dental procedures, mainly dentoalveolar surgery. Several preventive strategies to minimize the risk of developing MRONJ have been investigated. However, no investigation has been attempted to evaluate the therapeutic effect of local drug-delivery technology as a preventive strategy protocol. The aim of this study is to evaluate the efficacy of hydroxyapatite-containing doxycycline (HADOX) in rats with high-risk MRONJ development. All the rats used in this study were divided into seven groups. Six groups of rats out of seven were exposed to two different doses of antiresorptive drug therapy for four weeks before undergoing an upper incisor extraction. After 28 days, all the animals were euthanized, and the bone blocks were processed for histological and histomorphometrical evaluation. The histomorphometric analysis confirmed that newly formed bone (NFB) was present in all groups, with significant differences. NFB in the HADOX group treated with zoledronic acid at 4% showed (28.38; C.I. 22.29–34.48), which represents a significant increase compared to HA (15.69; C.I. 4.89–26.48) (p = 0.02). A similar pattern was observed in the HADOX group treated with zoledronic acid 8% ZA treatment (p = 0.001). Conclusions: HADOX did not inhibit any bone repair and reduced early inflammatory response. Hence, HADOX could promote bone healing in patients undergoing antiresorptive drug therapy.

Background: Bioceramic nanometer coatings have been regarded as potential substitutes for plasma-sprayed hydroxyapatite coatings, and the association with bone morphogenetic protein (BMP) is an attempt to achieve faster osseointegration to hasten oral rehabilitation. Objective: This study aimed to investigate the effect of recombinant human bone morphogenetic protein-7 (rhBMP-7) on the osseointegration of titanium implants coated with a thin film surface of hydroxyapatite (HA). Methods: Two implants (n = 24) were placed in each white New Zealand rabbits’ femur (n = 6). Implants were placed in the right femur after standard instrumentation (A and B) and in the left femur after an over-instrumentation (C and D), preventing bone-implant contact. The distal implants were installed associated with rhBMP-7 (groups B [regular instrumentation] and D [over-instrumentation]) and, also, in the absence of without BMP (control groups A [regular instrumentation] and C [over-instrumentation]). After 4 weeks, the animals were euthanized. The bone blocks containing the implants were embedded in methyl methacrylate and sectioned parallel to the long axis of the implant, which were analyzed by image segmentation. The data were analyzed using a nonparametric statistical method. Results: We observed that Group A had a mean bone formation of 35.6% compared to Group B, which had 48.6% (p > 0.05). Moreover, this group showed 28.3% of connective tissue compared to Group A, with 39.3%. In the over-instrumented groups, rhBMP-7 (Group D) showed an enhanced and significant increase in bone formation when compared with the group without rhBMP-7 (Group C). Conclusion: We concluded that the association of rhBMP-7 to thin nanostructure HA-coated implants promoted greater new bone area than the same implants in the absence of rhBMP-7, mainly in cases of over-instrumented implant sites.

Abstract Purpose: To analyze, histomorphologically, the influence of the geometry of nanostructured hydroxyapatite and alginate (HAn/Alg) composites in the initial phase of the bone repair. Methods: Fifteen rats were distributed to three groups: MiHA - bone defect filled with HAn/Alg microspheres; GrHA - bone defect filled with HAn/Alg granules; and DV - empty bone defect; evaluated after 15 days postoperatively. The experimental surgical model was the critical bone defect, ≅8.5 mm, in rat calvaria. After euthanasia the specimens were embedded in paraffin and stained with hematoxylin and eosin, picrosirius and Masson-Goldner’s trichrome. Results: The histomorphologic analysis showed, in the MiHA, deposition of osteoid matrix within some microspheres and circumjacent to the others, near the bone edges. In GrHA, the deposition of this matrix was scarce inside and adjacent to the granules. In these two groups, chronic granulomatous inflammation was noted, more evident in GrHA. In the DV, it was observed bone neoformation restricted to the bone edges and formation of connective tissue with reduced thickness in relation to the bone edges, throughout the defect. Conclusion: The geometry of the biomaterials was determinant in the tissue response, since the microspheres showed more favorable to the bone regeneration in relation to the granules.

Many biomaterials are used for Bone Morphogenetic Proteins (BMPs) delivery in bone tissue engineering. The BMP carrier system’s primary function is to hold these growth factors at the wound’s site for a prolonged time and provide initial support for cells to attach and elaborate the extracellular matrix for bone regeneration. This study aimed to evaluate the nanostructured carbonated hydroxyapatite microspheres (nCHA) as an rhBMP-2 carrier on rats calvaria. A total of fifteen male Wistar rats were randomly divided into three groups (n = 5): clot (control group), rhBMP-2 associated with collagen membrane (COL/rhBMP-2) or associated with the microspheres (nCHA/rhBMP-2). After 45 days, the calvaria defect samples were evaluated through histological, histomorphometric, and SR-µCT analyses to investigate new-formed bone and connective tissue volume densities. The descriptive histological analysis showed that nCHA/rhBMP-2 improved bone formation compared to other groups. These results were confirmed by histomorphometric and SR-µCT analysis that showed substantially defect area filling with a higher percentage of newly formed (36.24 ± 6.68) bone than those with the COL/rhBMP-2 (0.42 ± 0.40) and Clot (3.84 ± 4.57) (p < 0.05). The results showed that nCHA is an effective carrier for rhBMP-2 encouraging bone healing and an efficient alternative to collagen membrane for rhBMP-2 delivery.

Na região de Prado, sul da Bahia, Brasil, ocorre um importante depósito de caulim que tem sido explorado pela Mineração de Caulim Monte Pascoal desde 1992. O caulim destina-se principalmente à indústria do papel e para produção de catalisadores utilizados no craqueamaneto do petróleo. O depósito apresenta dois tipos distintos de minério. O caulim da base da sequência é de origem primária, qualidade superior e destina-se principalmente à indústria do papel. O caulim do topo é de origem sedimentar e de qualidade inferior, baixo grau de cristalinidade e alto teor de ferro e destina-se principalmente à indústria de catalisadores. O caulim foi submetido ao processo separação magnética e tratamento com agente redutor, ditionito de sódio visando elevar o índice de alvura e eliminar as impurezas mineralógicas presentes no caulim. As amostras foram caracterizadas através da difratometria de raios X, microscopia eletrônica de varredura e ressonância paramagnética eletrônica. Os espectros de RPE das amostras de caulim exibem ressonâncias paramagnéticas (g = 4,9 e 4,2) atribuídas ao Fe3+ em sítios de simetria octaedral na estrutura da caulinita substituindo o Al3+ e ressonância ferromagnéticas (g = 2,0) associadas a óxidos e/ou hidróxidos de ferro presentes em fases externas a caulinita.

Abstract The aim of this study is to evaluate the biocompatibility and osteoconductivity in surgical defects of sheep tibias filled with 1% strontium-containing nanostructured hydroxyapatite microspheres (SrHA), stoichiometric hydroxyapatite without strontium microspheres (HA), or blood clots. Santa Ines sheep were subjected to three perforations on the medial side of the left tibia. The biomaterials were characterized by X-ray Diffraction (XRD) and Fourier Transform Infrared (FTIR) before implantation and by X-Ray Microfluorescence (µFRX) and Scanning Electron Microscopy (SEM) after sheep tibias implantation. Surgical defects were filled with blood clots (control), SrHA (Group 1) or HA (Group 2). After 30 days, 5-µm bone blocks were obtained for histological evaluation, and the blocks obtained from 1 animal were embedded in methylmethacrylate for undecalcified sections. Mononuclear inflammatory infiltrate remained mild in all experimental groups. Giant cells were observed surrounding biomaterials particles of both groups and areas of bone formation were detected in close contact with biomaterials. All groups showed newly formed bone from the periphery to the center of the defects, which the control, HA and SrHA presented 36.4% (± 21.8), 31.2% (± 14.7) and 26.2% (± 12.9) of newly formed bone density, respectively, not presenting statistical differences. In addition, the connective tissue density did not show any significant between groups. The SrHA showing a higher volume density of biomaterial (51.2 ± 14.1) present in the defect compared to HA (32.6 ± 8.5) after 30 days (p = 0.03). Microspheres containing 1% SrHA or HA can be considered biocompatible, have osteoconductive properties and may be useful biomaterials for clinical applications.

In this work, the synthetic hydroxyapatite (HAP) was studied using different preparation routes to decrease the crystal size and to study the temperature effect on the HAP nano-sized hydroxyapatite crystallization. X-ray diffraction (XRD) analysis indicated that all samples were composed by crystalline and amorphous phases . The sample with greater quantity of amorphous phase (40% of total mass) was studied. The nano-sized hydroxyapatite powder was heated and studied at 300, 500, 700, 900 and 1150 °C. All samples were characterized by XRD and their XRD patterns refined using the Rietveld method. The crystallites presented an anisotropic form, being larger in the [001] direction. It was observed that the crystallite size increased continuously with the heating temperature and the eccentricity of the ellipsoidal shape changed from 2.75 at 300 °C to 1.94, 1.43, 1.04 and 1.00 respectively at 500, 700, 900 and 1150 °C. In order to better characterize the morphology of the HAP the samples were also examined using atomic force microscopy (AFM), infrared spectrometry (IR) and thermogravimetric analysis (TGA).

Top-down tissue engineering aims to produce functional tissues using biomaterials as scaffolds, thus providing cues for cell proliferation and differentiation. Conversely, the bottom-up approach aims to precondition cells to form modular tissues units (building-blocks) represented by spheroids. In spheroid culture, adult stem cells are responsible for their extracellular matrix synthesis, re-creating structures at the tissue level. Spheroids from adult stem cells can be considered as organoids, since stem cells recapitulate differentiation pathways and also represent a promising approach for identifying new molecular targets (biomarkers) for diagnosis and therapy. Currently, spheroids can be used for scaffold-free (developmental engineering) or scaffold-based approaches. The scaffold promotes better spatial organization of individual spheroids and provides a defined geometry for their 3D assembly in larger and complex tissues. Furthermore, spheroids exhibit potent angiogenic and vasculogenic capacity and serve as efficient vascularization units in porous scaffolds for bone tissue engineering. An automated combinatorial approach that integrates spheroids into scaffolds is starting to be investigated for macro-scale tissue biofabrication.

OBJETIVO: Os xenoenxertos são uma alternativa aos enxertos autogênicos e devem ser osteocondutores e biocompatíveis. Um estudo recente demonstrou que as características físicas e químicas das hidroxiapatitas encontradas no comércio não são condizentes com as descrições dos fabricantes. Este estudo avalia a biocompatibilidade e a osteocondução de um xenoenxerto derivado de osso medular bovino. MÉTODOS: Quinze coelhos adultos, pesando entre 2.500g e 3.000g cada um, foram anestesiados e submetidos a procedimentos cirúrgicos para criar três perfurações na tíbia, com 2 mm de diâmetro. Esses orifícios foram preenchidos com coágulo sanguineo, Osseus® e Bio-Oss®. Os animais foram sacrificados depois de 7, 14 e 28 dias. RESULTADOS: A análise histopatológica da interface de osso e biomaterial mostrou ligeira resposta inflamatória depois de 7 dias, que esteve ausente nos períodos subsequentes. Ambos os materiais foram parcialmente substituídos por tecido ósseo diretamente em contato com a superfície dos materiais, sem ocorrência de fibrose. CONCLUSÃO: Concluímos que ambos os materiais são biocompatíveis e absorvíveis, o que permite progressiva aposição óssea na superfície durante todo o período de avaliação.OBJECTIVE: Xenografts are an alternative to autogenous grafts and must be osteoconductive and biocompatible. A recent study has demonstrated that the physical and chemical characteristics of commercially available hydroxyapatites do not fit the manufacturers' descriptions. This study evaluates the biocompatibility and osteoconduction of a xenograft derived from bovine medullary bone. METHODS: Fifteen adult rabbits weighing between 2,500 g and 3,000 g each were anesthetized and subjected to surgical procedures to create three perforations in the tibia, with a diameter of 2 mm. These were filled with blood clot, Osseus® and Bio-Oss®. The animals were euthanized after 7, 14 and 28 days. RESULTS: Histopathological analysis of the interface between the bone and the biomaterial showed a slight inflammatory response after 7 days, which was absent in the subsequent periods. Both materials were partially replaced by bone tissue directly in contact with the surface of the materials, without the occurrence of fibrosis. CONCLUSION: We concluded that both materials are biocompatible and absorbable, allowing for progressive bone apposition on the surface throughout the evaluation period.

Synthetic porous ceramics can be used as three-dimensional scaffolds for bone repair. The aim of this work is to correlate process parameters with scaffolds structural characteristics. The factorial 2 level experimental design was chosen to study the effect of the Ca/P ratio (1.58 or 1.67), calcination temperature (1000 or 1150 °C) and porogen content (20 or 40%) on the calcium phosphate samples characteristics. The influence of such parameters was determined by X-ray diffraction, infrared spectroscopy and image analysis. It was observed that phase composition was basically a function of the Ca/P ratio of the raw material. The use of the porogen did not alter the hydroxyapatite (HA)/tricalcium phosphate (TCP) content, but induced changes in the relative content of TCP phase (alpha or beta). It is possible to design a porous sample with defined characteristics, and the model herein used can be considered as having a good predictive power.

First-principles modeling combined with experimental methods were used to study hydroxyapatite in which Sr2 substituted for Ca2. Detailed analyses of cation–oxygen bond distributions, cation–cation distances, and site 1–oxygen polyhedron twist angles were made in order to provide an atomic-scale interpretation of the observed structural modifications. Density functional theory periodic band-structure calculations indicate that the Ca2 Sr2닦⭮똪 induces strong local distortion on the hydroxyapatite lattice: the nearest neighbor Sr–O bond structures in both cationic sites are comparable to pure SrHA, while Sr induces more distortion at site 2 than site 1. Infrared vibrational spectroscopy (FTIR) and extended X-ray absorption fine structure (EXAFS) analysis suggest increasing lattice disorder and loss of OH with increasing Sr content. Rietveld refinement of synchrotron X-ray diffraction patterns shows a preference for the Ca1 site at Sr concentrations below 1 at.%. The ideal statistical occupancy ratio Sr2/Sr1 = 1.5 is achieved for ∼5 at.%; for higher Sr concentrations occupation of the Ca2 site is progressively preferred. [ABSTRACT FROM AUTHOR]