Incubation In Simulated Body Fluid Research Articles

Magnetic biocomposite scaffold based on decellularized tendon ECM and MNP-deposited halloysite nanotubes: physicochemical, thermal, rheological, mechanical and in vitro biological evaluations

The development of new three-dimensional biomaterials with advanced versatile properties is critical to the success of tissue engineering (TE) applications. Here, (a) bioactive decellularized tendon extracellular matrix (dECM) with a sol-gel transition feature at physiological temperature, (b) halloysite nanotubes (HNT) with known mechanical properties and bioactivity, and (c) magnetic nanoparticles (MNP) with superparamagnetic and osteogenic properties were combined to develop a new scaffold that could be used in prospective bone TE applications. Deposition of MNPs on HNTs resulted in magnetic nanostructures without agglomeration of MNPs. A completely cell-free, collagen- and glycosaminoglycan- rich dECM was obtained and characterized. dECM-based scaffolds incorporated with 1%, 2% and 4% MNP-HNT were analysed for their physical, chemical, and in vitro biological properties. Fourier-transform infrared spectroscopy, x-ray powder diffractometry and vibrating sample magnetometry analyses confirmed the presence of dECM, HNT and MNP in all scaffold types. The capacity to form apatite layer upon incubation in simulated body fluid revealed that dECM-MNP-HNT is a bioactive material. Combining dECM with MNP-HNT improved the thermal stability and compressive strength of the macroporous scaffolds upto 2% MNP-HNT. In vitro cytotoxicity and hemolysis experiments showed that the scaffolds were essentially biocompatible. Human bone marrow mesenchymal stem cells adhered and proliferated well on the macroporous constructs containing 1% and 2% MNP-HNT; and remained metabolically active for at least 21 d in vitro. Collectively, the findings support the idea that magnetic nanocomposite dECM scaffolds containing MNP-HNT could be a potential template for TE applications.

Hybrid Coatings Based on Polyvinylpyrrolidone/Polyethylene Glycol Enriched with Collagen and Hydroxyapatite: Incubation Studies and Evaluation of Mechanical and Physiochemical Properties.

Coating materials offers an intriguing solution for imparting inert implants with additional bioactive characteristics without changing underlying parameters such as mechanical strength. Metallic implants like endoprostheses or polymeric implants can be coated with a thin layer of bioactive film capable of stimulating bone-forming cells to proliferate or release a drug. However, irrespective of the final implantation site of such a coating biomaterial, it is necessary to conduct detailed mechanical and physicochemical in vitro analyses to determine its likely behavior under biological conditions. In this study, polymeric and composite coatings with hydroxyapatite obtained under UV light underwent incubation tests in four different artificial biological fluids: simulated body fluid (SBF), artificial saliva, Ringer's fluid, and water (as the reference fluid). The potentiometric and conductometric properties, sorption capacity, and degradation rate of the coatings were examined. Furthermore, their hardness, modulus of elasticity, and deformation were determined. It was demonstrated that the coatings remained stable in SBF liquid at a pH value of around 7.4. In artificial saliva, the greatest degradation of the polymer matrix (ranging between 36.19% and 39.79%) and chipping of hydroxyapatite in the composite coatings were observed. Additionally, the effect of ceramics on sorption capacity was determined, with lower capacity noted with higher HA additions. Moreover, the evaluation of surface morphology supported by elemental microanalysis confirmed the appearance of new apatite layers on the surface as a result of incubation in SBF. Ceramics also influenced mechanical aspects, increasing hardness and modulus of elasticity. For the polymer coatings, the value was 11.48 ± 0.61, while for the composite coating with 15% ceramics, it increased more than eightfold to a value of 93.31 ± 11.18 N/mm2. Based on the conducted studies, the effect of ceramics on the physicochemical as well as mechanical properties of the materials was determined, and their behavior in various biological fluids was evaluated. However, further studies, especially cytotoxicity analyses, are required to determine the potential use of the coatings as biomaterials.

Chitosan and Sodium Hyaluronate Hydrogels Supplemented with Bioglass for Bone Tissue Engineering.

The aim of the study was to produce biocomposites based on chitosan and sodium hyaluronate hydrogels supplemented with bioglasses obtained under different conditions (temperature, time) and to perform an in vitro evaluation of their cytocompatibility using both indirect and direct methods. Furthermore, the release of ions from the composites and the microstructure of the biocomposites before and after incubation in simulated body fluid were assessed. Tests on extracts from bioglasses and hydrogel biocomposites were performed on A549 epithelial cells, while MG63 osteoblast-like cells were tested in direct contact with the developed biomaterials. The immune response induced by the biomaterials was also evaluated. The experiments were carried out on both unstimulated and lipopolysaccharide (LPS) endotoxin-stimulated human peripheral blood cells in the presence of extracts of the biocomposites and their components. Extracts of the materials produced do not exhibit toxic effects on A549 cells, and do not increase the production of proinflammatory cytokines tumour necrosis factor alpha (TNF-α) and interleukin (IL-6) by blood cells in vitro. In direct contact with MG63 osteoblast-like cells, biocomposites containing the reference bioglass and those containing SrO are more cytocompatible than biocomposites with ZnO-doped bioglass. Using two testing approaches, the effects both of the potentially toxic agents released and of the surface of the tested materials on the cell condition were assessed. The results pave the way for the development of highly porous hydrogel-bioglass composite scaffolds for bone tissue engineering.

Biomimetic mineralization of platelet lysate/oxidized dextran cryogel as a macroporous 3D composite scaffold for bone repair

Scaffold development approaches using autologous sources for tissue repair are of great importance in obtaining bio-active/-compatible constructs. Platelet-rich plasma (PRP) containing various growth factors and platelet lysate (PL) derived from PRP are autologous products that have the potential to accelerate the tissue repair response by inducing a transient inflammatory event. Considering the regenerative capacity of PRP and PL, PRP/PL-based scaffolds are thought to hold great promise for tissue engineering as a natural source of autologous growth factors and a provider of mechanical support for cells. Here, a bio-mineralized PRP-based scaffold was developed using oxidized dextran (OD) and evaluated for future application in bone tissue engineering. Prepared PL/OD scaffolds were incubated in simulated body fluid (SBF) for 7, 14 and 21 d periods. Mineralized PL/OD scaffolds were characterized using Fourier transform infrared spectroscopy, x-ray diffraction spectroscopy, scanning electron microscopy (SEM), thermogravimetric analysis, porosity and compression tests. SEM and energy-dispersive x-ray spectroscopy analyses revealed mineral accumulation on the PL/OD scaffold as a result of SBF incubation. In vitro cytotoxicity and in vitro hemolysis tests revealed that the scaffolds were non-toxic and hemocompatible. Additionally, human osteoblasts (hOBs) exhibited good attachment and spreading behavior on the scaffolds and maintained their viability throughout the culture period. The alkaline phosphatase activity assay and calcium release results revealed that PL/OD scaffolds preserved the osteogenic properties of hOBs. Overall, findings suggest that mineralized PL/OD scaffold may be a promising scaffold for bone tissue engineering.

Surface modification of TiNb alloys by hydrothermal-precalcification treatment and its excellent in vitro osteogenic bioactivity

The superb comprehensive mechanical properties of titanium and its alloys make them ideal materials for artificial bone repair. However, the osseointegration between normal tissue and implant is lengthy due to the relatively poor surface osteogenic bioactivity of titanium-based materials. For the sake of accelerating the bone healing process, surface modification for the titanium- based implant is therefore of great necessity. In this paper, the novel [Formula: see text]-typed TiNb alloy with good biocompatibility was modified by a simple hydrothermal-precalcification method. The experimental results demonstrate that these hydrothermally prepared TiNb alloys were coated by a superhydrophilic Nb-doped TiO2 film with a hierarchical micro-nanostructure. After subsequent pre-calcification treatment, calcium and phosphorus species with low content were successfully introduced into the film without morphological change. The simulated body fluid (SBF) soaking experiments show that the TiNb alloy with the two-step surface treatment represented excellent in vitro bioactivity. After incubating in SBF for only 1 day, the TiNb alloy surface was thoroughly coated with a layer of newly formed spherical hydroxyapatite clusters. Comparatively, no obvious hydroxyapatite was deposited on the TiNb alloy surface with the one-step hydrothermal treatment even after five days of incubation in SBF. As a result, the hydrothermal–precalcification treatment is an ideal method to improve the osteogenic bioactivity of TiNb alloy, which can be further extended to surface modification of other titanium-based implant materials.

Influence of partial replacing of CaO by SrO on structure and bioactivity of calcium-phospho-silicate glasses

New alkali-free bioglasses of the ternary 60SiO2·5P2O5·35CaO parent glass wherein Ca is partially substituted by Sr (up to 10 mol%) were prepared by sol-gel method. The replacement of Ca with Sr induced the tendency to crystallisation of an apatite type phase. The bioactivity tested in vitro depends on Ca/Sr replacement degree, despite the fact that the theoretical network connectivity does not change with Ca/Sr substitution. After seven days of incubation in simulated body fluid, only on the samples with x ≥ 5 mol% SrO was developed a layer assumed of combined Ca/Sr composition of carbonated hydroxyapatite. Bactericidal activity was observed exclusively in case of strontium containing samples, for both Gram positive and negative bacteria. Structural effects of partial replacing of CaO by SrO were investigated by X-ray diffraction and FTIR analysis both before and after samples incubation in simulated body fluid.

Gold Nanoparticle‐Coated Bioceramics for Plasmonically Enhanced Molecule Detection via Surface‐Enhanced Raman Scattering

Herein, feasibility of plasmonically enhanced molecule detection via surface‐enhanced Raman scattering for ceramics that are commonly used as bone or tooth replacement materials is evaluated. Open cell foams of Bioglass 45S5, the commercial hydroxyapatite‐based product Bio‐Oss, and bioinert zirconia‐toughened‐alumina (ZTA) are coated with Au nanoparticles via colloidal deposition to introduce plasmonic effects. Depending on the pore size, gold‐functionalized plasmonic porous Bioglass shows effective Raman enhancement factor (eEF) up to , while depositing gold nanoparticles on Bio‐Oss and porous ZTA resulted in eEF of and respectively. The performance of the plasmonic porous bioceramics under simulated biological conditions is examined in situ in the biological medium fetal bovine serum (FBS) and during extended incubation in mineralizing simulated body fluid (SBF). Most notably, the plasmonic porous Bioglass still delivered an eEF around after 28 days of incubation in SBF, indicating promising stability in simulated biological conditions without significant difference in SBF bioactivity before and after Au deposition. Accordingly, the plasmonically enhanced porous bioceramics offer the possibility for real‐time and sensitive molecule detection at SBF and FBS conditions and can be further developed for sensing of specific biomarkers, for example, in the context of osseointegration of bone replacement materials.

Bioactive Glasses Enriched with Strontium or Zinc with Different Degrees of Structural Order as Components of Chitosan-Based Composite Scaffolds for Bone Tissue Engineering.

The development of innovative biomaterials with improved integration with bone tissue and stimulating regeneration processes is necessary. Here, we evaluate the usefulness of bioactive glasses from the SiO2-P2O5-CaO system enriched with 2 wt.% SrO or ZnO in the manufacturing of chitosan-based scaffolds. Bioglasses produced using the sol-gel method were subjected to thermal treatment in different regimes. Chitosan/bioglass composites were produced with a weight ratio. Bioglasses were evaluated via TG-DTA, FTIR, and SEM-EDS before and after incubation in simulated body fluid (SBF). The release of ions was tested. The cytocompatibility of the composites in contact with MG63 osteoblast-like cells was evaluated. The results showed that the presence of the crystalline phase decreased from 41.2-44.8% for nonmodified bioglasses to 24.2-24.3% for those modified with ZnO and 22.0-24.2% for those modified with SrO. The samples released Ca2+, Zn2+, and/or Sr2+ ions and were bioactive according to the SBF test. The highest cytocompatibility was observed for the composites containing nonmodified bioglasses, followed by those enriched with SrO bioglasses. The least cytocompatible were the composites containing ZnO bioglasses that released the highest amount of Zn2+ ions (0.58 ± 0.07 mL/g); however, those that released 0.38 ± 0.04 mL/g were characterised by acceptable cytocompatibility. The study confirmed that it is feasible to control the biological performance of chitosan/bioglass composites by adjusting the composition and heat treatment parameters of bioglasses.

Evaluating the bioactivity of endodontic sealers with respect to their thermo-nanomechanical properties

Abstract Physico-chemical and mechanical features of endodontic sealers are essential functional properties involved in their sealing ability, osteoconductivity, and biocompatibility. Four different root canal sealers (Ceraseal, MTA Fillapex, AH Plus and Sealapex) were investigated in this study in order to evaluate the thermal and nanomechanical features in relation to their bioactivity potential. The nano-hardness values respected the descendent trend: HCeraseal >HSealapex >HMTA >HAHPlus, being influenced by the percent of the inorganic component in the samples, identified as residual mass in TGA/DTG (Thermogravimetrical Analysis/ Derivative Thermogravimetry) thermograms. Based on electrochemical measurement in SBF (Simulated Body Fluid) and surface investigations by SEM/EDX, we found that the bioactivity potential decreased in the following order: Ceraseal > MTA Fillapex >AH Plus >Sealapex. The highest bioactivity in the case of Ceraseal was evidenced in terms of apatite-like spherulites observed to cover the surface after 28 days incubation in SBF, and Ca/P ratio 1.71, along with the dynamic release and consumption of Ca2+ and PO3− in SBF. On the opposite side, lack of mineralization was noticed on the surface of Sealapex sample. Knowledge about the proper thermo-mechanical, biological and physico-chemical properties of the root canal materials is important in order to allow a correct material selection, either as premixed or two-component formulation, requiring good sealing or immediate therapeutic effect.

Development, physicochemical characterization and in-vitro biocompatibility study of dromedary camel dentine derived hydroxyapatite for bone repair.

This study aimed to produce hydroxyapatite from the dentine portion of camel teeth using a defatting and deproteinizing procedure and characterize its physicochemical and biocompatibility properties. Biowaste such as waste camel teeth is a valuable source of hydroxyapatite, the main inorganic constituent of human bone and teeth which is frequently used as bone grafts in the biomedical field. Fourier Transform infrared (FTIR), and micro-Raman spectroscopy confirmed the functional groups as-sociated with hydroxyapatite. X-ray diffraction (XRD) studies showed camel dentine-derived hydroxyapatite (CDHA) corresponded with hydroxyapatite spectra. Scanning electron micros-copy (SEM) demonstrated the presence of dentinal tubules measuring from 1.69-2.91 µm. The inorganic phases of CDHA were primarily constituted of calcium and phosphorus, with trace levels of sodium, magnesium, potassium, and strontium, according to energy dispersive X-ray analysis (EDX) and inductively coupled plasma mass spectrometry (ICP-MS). After 28 days of incubation in simulated body fluid (SBF), the pH of the CDHA scaffold elevated to 9.2. in-vitro biocompatibility studies showed that the CDHA enabled Saos-2 cells to proliferate and express the bone marker osteonectin after 14 days of culture. For applications such as bone augmentation and filling bone gaps, CDHA offers a promising material. However, to evaluate the clinical feasibility of the CDHA, further in-vivo studies are required.

Cold hydrostatic sintering of 45S5 bioactive glass

Fully amorphous 45S5 bioactive glass pellets were successfully consolidated at room temperature using cold hydrostatic sintering (CHS) and their acellular bioactivity was determined by immersion test in simulated body fluid (SBF). Crystalline 45S5 BG samples prepared using conventional sintering (TS) at 1050 °C were also tested. Preliminary results indicated the formation of hydroxyapatite (HAP) on the surfaces of both samples after immersion in SBF, which was confirmed using SEM, FTIR, and XRD analyses. The amount of hydroxyapatite increases by increasing the SBF immersion time. Results revealed that the amount of HAP crystalline phase was similar in both samples after 14 days of incubation in SBF (i.e., 23% in TS and 25% in CHS samples). The pellets prepared by TS resulted in a relative density of 95.6% and a hardness of about 3.5 GPa. On the other hand, CHS samples had a hardness of 1.2 GPa and a relative density of 75.1%, indicating the presence of a porous structure that can be beneficial for the interaction of the material with proteins and cells (both in vitro and in vivo).

Characteristics of Hybrid Bioglass-Chitosan Coatings on the Plasma Activated PEEK Polymer

Polyetheretherketone (PEEK) is a biocompatible, chemically and physically stable radiolucent polymer that exhibits a similar elastic modulus to the normal human bone, making it an attractive orthopedic implant material. However, PEEK is biologically inert, preventing strong enough bonding with the surrounding bone tissue when implanted in vivo. Surface modification and composite preparation are the two main strategies for the improvement of the bioactivity of PEEK. In this study, the plasma activated PEEK surfaces with the embedded bioglass, chitosan, and bioglass-chitosan mixed layers applying from the solution dip-coating technique were investigated. The most prominent factors affecting the coating biocompatibility are strictly connected with the composition of its outer surface (its charge and functional groups), hydrophilic-hydrophobic character, wettability and surface free energy, and topography (size of pores/substructures, roughness, stiffness), as well as the personal characteristics of the patient. The obtained surfaces were examined in terms of wettability and surface-free energy changes. Additionally, FTIR (Fourier Transformation Infrared Spectrometry) and SIMS (Secondary Ion Mass Spectrometry) were applied to establish and control the coating composition. Simultaneously the structure of coatings was visualized with the aid of SEM (Scanning Electron Microscopy). Finally, the obtained systems were incubated in SBF (Simulated Body Fluid) to verify the modifications' influence on the bioactivity/biocompatibility of the PEEK surface. Different structures with variable compositions, as well as changes of the wettability, were observed depending on the applied modification. In addition, the incubation in SBF suggested that the bioglass-chitosan ratio influenced the formation of apatite-like structures on the modified PEEK surfaces.

Zinc- and Copper-Doped Mesoporous Borate Bioactive Glasses: Promising Additives for Potential Use in Skin Wound Healing Applications.

In this study, zinc (Zn)- and copper (Cu)-doped 13-93B3 borate mesoporous bioactive glasses (MBGs) were successfully synthesized using nitrate precursors in the presence of Pluronic P123. We benefited from computational approaches for predicting and confirming the experimental findings. The changes in the dynamic surface tension (SFT) of simulated body fluid (SBF) were investigated using the Du Noüy ring method to shed light on the mineralization process of hydroxyapatite (HAp) on the glass surface. The obtained MBGs were in a glassy state before incubation in SBF. The formation of an apatite-like layer on the SBF-incubated borate glasses was investigated by X-ray diffraction (XRD) and scanning electron microscopy (SEM). The incorporation of Zn and Cu into the basic composition of 13-93B3 glass led to changes in the glass transition temperature (Tg) (773 to 556 °C), particle size (373 to 64 nm), zeta potential (-12 to -26 mV), and specific surface area (SBET) (54 to 123 m2/g). Based on the K-means algorithm and chi-square automatic interaction detection (CHAID) tree, we found that the SFT of SBF is an important factor for the prediction and confirmation of the HAp mineralization process on the glasses. Furthermore, we proposed a simple calculation, based on SFT variation, to quantify the bioactivity of MBGs. The doped and dopant-free borate MBGs could enhance the proliferation of mouse fibroblast L929 cells at a concentration of 0.5 mg/mL. These glasses also induced very low hemolysis (<5%), confirming good compatibility with red blood cells. The results of the antibacterial test revealed that all the samples could significantly decrease the viability of Pseudomonas aeruginosa. In summary, we showed that Cu-/Zn-doped borate MBGs can be fabricated using a cost-effective method and also show promise for wound healing/skin tissue engineering applications, as especially supported by the cell test with fibroblasts, good compatibility with blood, and antibacterial properties.

Protein‐Coated Biodegradable Gas‐Stabilizing Nanoparticles for Cancer Therapy and Diagnosis Using Focused Ultrasound

AbstractSurface‐engineered hydrophobic nanoparticles that can stabilize small gas pockets on their surfaces (i.e., gas‐stabilizing nanoparticles, GSNs) have been recently shown to be excellent contrast and cavitation agents for ultrasound theranostics. However, previously developed GSNs are not biodegradable, which limits their clinical translation potential. Here the development of biodegradable GSNs is shown by coating hydrophobically modified mesoporous silica nanoparticles with different protein solutions. It is found that these novel GSNs retain strong cavitation activity while rapidly degrading in simulated body fluid (SBF) or in vivo in days or a few weeks, respectively. Interestingly, GSNs coated with other stabilizing layers, Pluronic F127 polymer or phospholipids, demonstrated significantly slower degradation rates with only partial degradation even after a month of incubation in SBF. Next, it is shown that these biodegradable GSNs can be used to ablate tumor xenografts at lower ultrasound intensities, thus avoiding the side effects of high‐intensity ultrasound. Finally, it is shown that only tumors treated with GSNs and ultrasound can specifically enrich for circulating tumor DNA, which will improve liquid biopsies for understanding tumor heterogeneity and treatment response. Overall, this study details a simple yet effective method for preparing biodegradable GSNs with broad potential for applications in cancer diagnosis and therapy.

Extraction of Hydroxyapatite from Camel Bone for Bone Tissue Engineering Application.

Waste tissues such as mammalian bone are a valuable source from which to extract hydroxyapatite. Camel bone-based hydroxyapatite (CBHA) was extracted from the femur of camel bones using a defatting and deproteinization procedure. The extracted CBHA was mechanically, chemically, physically, morphologically and structurally characterized. Fourier-Transform Infra-Red (FTIR) spectra, Micro-Raman, and X-ray diffraction analysis confirmed successful extraction of hydroxyapatite. The mechanical properties of the CBHA scaffold were measured using a Universal Instron compression tester. Scanning electron microscopy showed the presence of a characteristic interconnected porous architecture with pore diameter ranging from 50-600 µm and micro-computer tomography (Micro-CT) analysis identified a mean porosity of 73.93. Thermogravimetric analysis showed that the CBHA was stable up to 1000 °C and lost only 1.435% of its weight. Inductively coupled plasma-mass spectrometry (ICP-MS) and Energy-dispersive-X-ray (EDX) analysis demonstrated the presence of significant amounts of calcium and phosphorus and trace ions of sodium, magnesium, zinc, lead and strontium. Following 21 days of incubation in simulated body fluid (SBF), the pH fluctuated between 10-10.45 and a gradual increase in weight loss was observed. In conclusion, the extracted CBHA is a promising material for future use in bone tissue regeneration applications.

Mineral-induced bubbling effect and biomineralization as strategies to create highly porous and bioactive scaffolds for dentin tissue engineering.

The objective of the study was to assess the biological and mechanical characteristics of chitosan-based scaffolds enriched by mineral phases and biomineralized in simulated body fluid (SBF) as a possible biomaterial for dentin regeneration. Thus, porous chitosan scaffolds were prepared by the mineral-induced bubbling-effect technique and subjected to biomineralization to create biomimetic scaffolds for dentin tissue engineering. Suspensions containing calcium hydroxide, nanohydroxyapatite, or β-tricalcium phosphate were added to the chitosan (CH) solution and subjected to gradual freezing and freeze-drying to obtain CHCa, CHnHA, and CHβTCP porous scaffolds, respectively, by the bubbling effect. Then, scaffolds were incubated in SBF for 5 days at 37°C, under constant stirring, to promote calcium-phosphate (CaP) biomineralization. Scanning electron microscopy revealed increased pore size and porosity degree on mineral-containing scaffolds, with CHCa and CHnHA presenting as round, well-distributed, and with an interconnected pore network. Nevertheless, incubation in SBF disrupted the porous architecture, except for CHCaSBF , leading to the deposition of CaP coverage, confirmed by Fourier Transform Infrared Spectroscopy analyses. All mineral-containing and SBF-treated formulations presented controlled degradation profiles and released calcium throughout 28 days. When human dental pulp cells (HDPCs) were seeded onto scaffold structures, the porous and interconnected architecture of CHCa, CHnHA, and CHCaSBF allowed cells to infiltrate and spread throughout the scaffold structure, whereas in other formulations cells were dispersed or agglomerated. It was possible to determine a positive effect on cell proliferation and odontogenic differentiation for mineral-containing formulations, intensely improved by biomineralization. A significant increase in mineralized matrix deposition (by 8.4 to 18.9 times) was observed for CHCaSBF , CHnHASBF , and CHβTCPSBF in comparison with plain CH. The bioactive effect on odontoblastic marker expression (ALP activity and mineralized matrix) was also observed for HDPCs continuously cultivated with conditioned medium obtained from scaffolds. Therefore, biomineralization of chitosan scaffolds containing different mineral phases was responsible for increasing the capacity for mineralized matrix deposition by pulpal cells, with potential for use in dentin tissue engineering.

Osteogenic Potential of Magnesium (Mg)-Doped Multicomponent Bioactive Glass: In Vitro and In Vivo Animal Studies

The use of bioactive glasses (BGs) has been quite fruitful in hard tissue engineering due to the capability of these materials to bond to living bone. In this work, a melt-derived magnesium (Mg)-doped BG (composition: 45SiO2–3P2O5–26CaO–15Na2O–7MgO–4K2O (mol.%)) was synthesized for being used in bone reconstruction. The prepared BGs were then manufactured as three-dimensional (3D) scaffolds by using the sponge replica approach. The microstructure of the samples was assessed by X-ray diffraction (XRD) and the surface morphology was observed by using scanning electron microscopy (SEM). The in vitro bioactivity and the release of osteo-stimulatory Mg2+ ions from the prepared samples were investigated over 7 days of incubation in simulated body fluids (SBF). In vitro cellular analyses revealed the compatibility of the Mg-doped BGs with human osteosarcoma cells (MG-63 cell line). Moreover, the Mg-doped BGs could induce bone nodule formation in vitro and improve the migratory ability of human umbilical vein endothelial cells (HUVECs). In vivo osteogenic capacity was further evaluated by implanting the BG-derived scaffolds into surgically-created critical-size bone defects in rats. Histological and immunohistological observations revealed an appropriate bone regeneration in the animals receiving the glass-based scaffolds after 12 weeks of surgery. In conclusion, our study indicates the effectiveness of the Mg-doped BGs in stimulating osteogenesis in both in vitro and in vivo conditions.

Biological and bioactivity assessment of dextran nanocomposite hydrogel for bone regeneration.

Insufficient biological and bioactive properties of dextran hydrogels limit their applications as promising scaffolds for tissue engineering. We developed nanocomposite dextran hydrogels comprised of bioactive glass (nBGC: 64% SiO2, 31% CaO, 5% P2O5) nanoparticles with an average particle size of 77nm using a chemical crosslinking of dextran chains to form 3D hydrogel networks. In the current study; bioactivity of the obtained nanocomposite hydrogels was evaluated through the formation of apatite crystal structures after the incubation in simulated body fluid (SBF) at various submersion periods and nBGC content. The scanning electron microscopy (SEM) micrographs represented an enhanced hydroxyapatite formation on the cross section of nanocomposite comprising of nBGC content from 2 to 8 (% by wt). Biomineralization results of Dex-8 (% by wt) composite during 7, 14 and 28days immersion indicated the apatite layer formation and the growth of apatite crystal size on the surface and cross section of the nanocomposite. Moreover, MTT assessments indicated that human osteosarcoma cells (SaOS-2) were able to adhere and spread within the dextran hydrogels reinforced with the bioactive glass nanoparticles. With regard to enhanced bioactivity and biocompatibility, the developed dextran-nBGC hydrogel could be considered as a suitable candidate for bone tissue engineering application.

MSLA-based 3D printing of acrylated epoxidized soybean oil - nano-hydroxyapatite composites for bone repair

Structural bone allografts are used to treat critically sized segmental bone defects (CSBDs) as such defects are too large to heal naturally. Development of biomaterials with competent mechanical properties that can also facilitate new bone formation is a major challenge for CSBD repair. 3D printed synthetic bone grafts are a possible alternative to structural allografts if engineered to provide appropriate structure with sufficient mechanical properties. In this work, we fabricated a set of novel nanocomposite biomaterials consisting of acrylated epoxidized soybean oil (AESO), polyethylene glycol diacrylate (PEGDA) and nanohydroxyapatite (nHA) by using masked stereolithography (mSLA)-based 3D printing.The nanocomposite inks possess suitable rheological properties and good printability to print complex, anatomically-precise, ‘by design’ grafts. The addition of nHA to the AESO/PEGDA resin improved the tensile strength and fracture toughness of the mSLA printed nanocomposites, presumably due to small-scale reinforcement. By adding 10 vol% nHA, tensile strength, modulus and fracture toughness (KIc) were increased to 30.8 ± 1.2 MPa (58% increase), 1984.4 ± 126.7 MPa (144% increase) and 0.6 ± 0.1 MPa·m1/2 (42% increase), respectively (relative to the pure resin). The nanocomposites did not demonstrate significant hydrolytic, enzymatic or oxidative degradation when incubated for 28 days, assuring chemical and mechanical stability at early stages of implantation. Apatite nucleated and covered the nanocomposite surfaces within 7 days of incubation in simulated body fluid. Good viability and proliferation of differentiated MC3T3-E1 osteoblasts were also observed on the nanocomposites. Taken all together, our nanocomposites demonstrate excellent bone-bioactivity and potential for bone defect repair.

The Construction of Multi-Incorporated Polylactic Composite Nanofibrous Scaffold for the Potential Applications in Bone Tissue Regeneration.

To improve the bone regeneration ability of pure polymer, varieties of bioactive components were incorporated to a biomolecular scaffold with different structures. In this study, polysilsesquioxane (POSS), pearl powder and dexamethasone loaded porous carbon nanofibers (DEX@PCNFs) were incorporated into polylactic (PLA) nanofibrous scaffold via electrospinning for the application of bone tissue regeneration. The morphology observation showed that the nanofibers were well formed through electrospinning process. The mineralization test of incubation in simulated body fluid (SBF) revealed that POSS incorporated scaffold obtained faster hydroxyapatite depositing ability than pristine PLA nanofibers. Importantly, benefitting from the bioactive components of pearl powder like bone morphogenetic protein (BMP), bone mesenchymal stem cells (BMSCs) cultured on the composite scaffold presented higher proliferation rate. In addition, by further incorporating with DEX@PCNFs, the alkaline phosphatase (ALP) level and calcium deposition were a little higher based on pearl powder. Consequently, the novel POSS, pearl powder and DEX@PCNFs multi-incorporated PLA nanofibrous scaffold can provide better ability to enhance the biocompatibility and accelerate osteogenic differentiation of BMSCs, which has potential applications in bone tissue regeneration.