Synthesis of Submicrometric Chitosan Particles Loaded with Calcium Phosphate for Biomedical Applications.

Chitosan particles loaded with dibasic calcium phosphate anhydrous (DCPA) is a promising strategy for combining antimicrobial and osteoconduction properties in regenerative medicine. However, mostly micrometer-sized particles have been reported in the literature, limiting their use and reducing their effect in the biomedical field. We have recently overcome this limitation by developing submicrometer-sized particles with electrospray technique. The objective of this study was to understand how the process parameters control the size and properties of submicrometer chitosan particles loaded with DCPA. Solutions of 10 mg/mL chitosan and 2.5 mg/mL DCPA in a 90% acetic acid were electrosprayed under three distinct flow rate conditions: 0.2, 0.5, and 1.0 mL/h. The particles were crosslinked in a glutaraldehyde atmosphere and characterized in terms of their morphology, inorganic content, zeta potential, and minimum inhibitory concentration (MIC) against S. mutans. All conditions showed particles with two similar morphologies: one small-sized with a spherical shape and another larger-sized with a bi-concave shape. All generated a broad particle size distribution, with a similar mean size of ~ 235 nm. The addition of DCPA decreased the zeta potential for all the samples, but it was above 30 mV, indicating a low aggregation potential. The lower flow rate showed the worst efficacy for DCPA incorporation. Antimicrobial activity was greater in chitosan/DCPA particles with flow rate of 0.5 mL/h. It can be concluded that the flow rate of 0.5 mL/h presents the best compromise solution in terms of morphology, zeta potential, MIC, and inorganic content.

Effects of the crosslinking of chitosan/DCPA particles in the antimicrobial and mechanical properties of dental restorative composites.

The development of restorative materials containing antibacterial agents is an alternative to reduce the progression of caries lesions. OBJECTIVE: to compare the influence of the degree of crosslinking of chitosan particles loaded with dibasic calcium phosphate (DCPA) on the mechanical properties, degree of conversion (DC), and antimicrobial properties of experimental composites. METHODS: Chitosan/DCPA particles were synthesized by the electrospraying, crosslinked by 0, 8, or 16 h in glutaraldehyde, and characterized by zeta potential and minimum inhibitory concentration (MIC) against S. mutans. Experimental resin composites of Bis-GMA and TEGDMA and 59.5% of barium glass were synthesized, chitosan/DCPA particles were added at 0 or 0.5 wt% with the different crosslinking time. The materials were subject to DC analysis, three-point bending test at 24 h and 7 days, and antimicrobial assays. Data were submitted to one-way ANOVA and Tukey test (α = 0.05). RESULTS: The particles with longer crosslinking time presented higher zeta potential and MIC, and the composite containing these particles showed significantly higher biofilm inhibition than the control group. The other two groups were similar to each other and the control. The composite containing particles with 88 h crosslinking time showed the lowest flexural strength at 7 days in water, and materials with non-crosslinked particles and longer crosslinking time presented flexural strength similar to control. The flexural modulus and DC showed no statistical difference among groups. SIGNIFICANCE: composite resin containing 0.5% chitosan/DCPA particles crosslinked by 16 h showed a reduction of biofilm formation without affecting the mechanical properties in relation to the control.

Physical and Biological Properties of a Chitosan Hydrogel Scaffold Associated to Photobiomodulation Therapy for Dental Pulp Regeneration: An In Vitro and In Vivo Study.

BACKGROUND: The regeneration of dental pulp, especially in cases of pulp death of immature teeth, is the goal of the regenerative endodontic procedures (REPs) that are based on tissue engineering principles, consisting of stem cells, growth factors, and scaffolds. Photobiomodulation therapy (PBMT) showed to improve dental pulp regeneration through cell homing approaches in preclinical studies and has been proposed as the fourth element of tissue engineering. However, when a blood clot was used as a scaffold in one of these previous studies, only 30% of success was achieved. The authors pointed out the instability of the blood clot as the regeneration shortcoming. Then, to circumvent this problem, a new scaffold was developed to be applied with the blood clot. The hypothesis of the present study was that an experimental injectable chitosan hydrogel would facilitate the three-dimensional spatial organization of endogenous stem cells in dental pulp regeneration with no interference on the positive influence of PBMT. METHODS: For the in vitro analysis, stem cells from the apical papilla (SCAPs) were characterized by flow cytometry and applied in the chitosan scaffold for evaluating adhesion, migration, and proliferation. For the in vivo analysis, the chitosan scaffold was applied in a rodent orthotopic dental pulp regeneration model under the influence of PBMT (660 nm; power output of 20 mW, beam area of 0.028 cm2, and energy density of 5 J/cm2). RESULTS: The scaffold tested in this study allowed significantly higher viability, proliferation, and migration of SCAPs in vitro when PBMT was applied, especially with the energy density of 5 J/cm2. These results were in consonance to those of the in vivo data, where pulp-like tissue formation was observed inside the root canal. CONCLUSION: Chitosan hydrogel when applied with a blood clot and PBMT could in the future improve previous results of dental pulp regeneration through cell homing approaches.

Vacancy-Induced Visible Light-Driven Fluorescence in Toxic Ion-Free Resorbable Magnetic Calcium Phosphates for Cell Imaging Applications.

Multifunctional nanosized particles are very beneficial in the field of biomedicine. Bioactive and highly biocompatible calcium phosphate (CaP) nanoparticles (∼50 nm) exhibiting both superparamagnetic and fluorescence properties were synthesized by incorporating dual ions (Fe3+ and Sr2+) in HAp (hydroxyapatite) [Ca10(PO4)6(OH)2]. Insertion of Fe3+ creates oxygen vacancies at the PO43- site, thereby destabilizing the structure. Thus, in order to maintain the structural stability, Sr2+ has been incorporated. This incorporation of Sr2+ leads to an intense emission at 550 nm. HAp nanoparticles when subjected to thermal treatment (800 °C) transform to ß-TCP, exhibiting emission at 710 nm due to the emergence of an intermediate band. Moreover, these nanoparticles exhibit fluorescence in visible light when compared to the other UV and IR fluorescence excitation sources which could damage the tissues. The synthesis involving the combination of ultrasound and microwave techniques resulted in the distribution of Fe3+ in the interstitial sites of CaP, which is responsible for the excellent fluorescent properties. Moreover, thermally treated CaP becomes superparamagnetic, without affecting the desired optical properties. The bioactive, biocompatible, magnetic, and fluorescent properties of this resorbable CaP which is free from toxic heavy metals (Eu, Gd, etc.) could help in overcoming the long-term cytotoxicity. This could also be useful in tracking the location of the nanoparticles during drug delivery and magnetic hyperthermia. The bioactive fluorescent CaP nanoparticle helps in monitoring the bone growth and in addition, it could be employed in cell imaging applications. The in vitro MCF-7 imaging using the nanoparticles after 24 h of uptake at 465 nm evidences the bioimaging capability of the prepared nanoparticles. The reproducibility of the defect level is essential for the defect-induced emission properties. The synthesis of nontoxic fluorescent CaP is highly reproducible with the present synthesis method. Hence, it could be safely employed in various biomedical applications.

Disinfection of 3D-printed protective face shield during COVID-19 pandemic.

This study assessed the disinfection using 70% ethanol; H2O2-quaternary ammonium salt mixture; 0.1% sodium hypochlorite and autoclaving of four 3D-printed face shields with different designs, visor materials; and visor thickness (0.5-0.75 mm). We also investigated their clinical suitability by applying a questionnaire to health workers (HW) who used them. Each type of disinfection was done 40 times on each type of mask without physical damage. In contrast, autoclaving led to appreciable damage.

Development of Epidermal Equivalent from Electrospun Synthetic Polymers for In Vitro Irritation/Corrosion Testing.

The development of products for topical applications requires analyses of their skin effects before they are destined for the market. At present, the ban on animal use in several tests makes the search for in vitro models (such as artificial skin) necessary to characterize the risks involved. In this work, tissue engineering concepts were used to manufacture collagen-free three-dimensional scaffolds for cell growth and proliferation. Two different human skin models-reconstructed human epidermis and full-thickness skin-were developed from electrospun scaffolds using synthetic polymers such as polyethylene terephthalate, polybutylene terephthalate, and nylon 6/6. After the construction of these models, their histology was analyzed by H&E staining and immunohistochemistry. The results revealed a reconstructed epidermal tissue, duly stratified, obtained from the nylon scaffold. In this model, the presence of proteins involved in the epidermis stratification process (cytokeratin 14, cytokeratin 10, involucrin, and loricrin) was confirmed by immunohistochemistry and Western blot analysis. The nylon reconstructed human epidermis model's applicability was evaluated as a platform to perform irritation and corrosion tests. Our results demonstrated that this model is a promising platform to assess the potential of dermal irritation/corrosion of chemical products.

Effect of ozone therapy on wound healing in the buccal mucosa of rats.

OBJECTIVE: The objective of this study was to evaluate the effects of ozone therapy on wound healing formed experimentally in the oral cavity of rats. DESIGN: Two surgical wounds were generated on the cheeks of 24 Wistar rats, bilaterally. Half of the animals were submitted to ozone therapy on both wounds (experimental group) and the other half received no treatment (control group). In the experimental group, wounds were exposed to ozone gas 1, 2 or 3 (60 µg/mL) times. Evaluation of wound healing of the buccal mucosa was followed for 1, 3 and 7 days. The distribution of neutrophils, fibroplasia and angiogenesis were analyzed. Samples were classified in a healing numerical scale according to the inflammatory intensity. Data were submitted to Mann-Whitney and Kruskal-Wallis tests (α = 0.05). RESULTS: On day 1, wounds were similar in both groups, lesions were open and bloody with slightly minor bleeding in the ozone therapy group. On day 3, the group with ozone therapy was almost all refurbished and with higher angiogenesis, while the control group still had more bloody points and lower blood vessels. On day 7, both wounds were remodeled, with higher fibroplasia in the group that received ozone therapy. CONCLUSION: It can be concluded that ozone therapy was effective in improving angiogenesis and fibroblasts count in the buccal mucosa of rats.

Bromelain immobilization in cellulose triacetate nanofiber membranes from sugarcane bagasse by electrospinning technique.

Cellulose triacetate (CTAB) synthesized by cellulose extracted from sugarcane bagasse, and commercial cellulose acetate (CA) were used to produce nanofiber membranes contained bromelain by electrospinning technique. About 1.3 g of cellulose acetate per gram of bagasse were obtained, and both CTAB and CA was characterized by analysis of Fourier Transform Infrared Spectroscopy (FTIR) and Differential Scanning Calorimetry (DSC). The nanofiber membranes were produced by electrospinning process testing the following conditions: voltage 25 kV, flow rate 4 mL/h and distance 10 cm, using acetone/ dimethylformamide (DMF) (85:15 m/ m) to 15% cellulose triacetate (70% CA + 30% CTAB) or CA solutions. Scanning Electron Microscopy (SEM) was used to nanofiber membranes characterization. Bromelain was immobilized on the nanofiber membranes by crosslinking with glutaraldehyde and directly in the electrospinning step, the highest activity recovery was about 675% and in vitro controlled release tests were performed to semi-quantitatively evaluate the release of the enzyme bromelain thus demonstrating complete release process in 3 days.

Thermally Modified Iron-Inserted Calcium Phosphate for Magnetic Hyperthermia in an Acceptable Alternating Magnetic Field.

Magnetic hyperthermia treatment using calcium phosphate nanoparticles is an evolutionary choice because of its excellent biocompatibility. In the present work, Fe3+ is incorporated into HAp nanoparticles by thermal treatment at various temperatures. Induction heating was examined within the threshold H f value of 4.58 × 106 kA m-1 s-1 (H is the strength of alternating magnetic field and f is the operating frequency) and sample concentration of 10 mg/mL. The temperature-dependent structural modifications are well correlated with the morphological, surface charge, and magnetic properties. Surface charge changes from +10 mV to -11 mV upon sintering because of the diffusion of iron in the HAp lattice. The saturation magnetization has been achieved by sintering the nanoparticles at 400 and 600 °C, which has led to the specific absorption rate of 12.2 and 37.2 W/g, respectively. Achievement of the hyperthermia temperature (42 °C) within 4 min is significant when compared with the existing magnetic calcium phosphate nanoparticles. The systematic investigation reveals that the HAp nanoparticles partially stabilized with FeOOH and biocompatible α-Fe2O3 exhibit excellent induction heating. In vitro tests confirmed the samples are highly hemocompatible. The importance of the present work lies in HAp nanoparticles exhibiting induction heating without compromising the factors such as H f value, low sample concentration, and reduced duration of applied field.

Modular Design of Programmable Mechanofluorescent DNA Hydrogels.

Mechanosensing systems are ubiquitous in nature and control many functions from cell spreading to wound healing. Biologic systems typically rely on supramolecular transformations and secondary reporter systems to sense weak forces. By contrast, synthetic mechanosensitive materials often use covalent transformations of chromophores, serving both as force sensor and reporter, which hinders orthogonal engineering of their sensitivity, response and modularity. Here, we introduce FRET-based, rationally tunable DNA tension probes into macroscopic 3D all-DNA hydrogels to prepare mechanofluorescent materials with programmable sacrificial bonds and stress relaxation. This design addresses current limitations of mechanochromic system by offering spatiotemporal resolution, as well as quantitative and modular force sensing in soft hydrogels. The programmable force probe design further grants temporal control over the recovery of the mechanofluorescence during stress relaxation, enabling reversible and irreversible strain sensing. We show proof-of-concept applications to study strain fields in composites and to visualize freezing-induced strain patterns in homogeneous hydrogels.

A fast degrading PLLA composite with a high content of functionalized octacalcium phosphate mineral phase induces stem cells differentiation.

Octacalcium phosphate (OCP) was synthesized yielding a combination of OCP and hydroxyapatite (HA) with a ratio of 90:10 (OCP/HA). A method was developed to functionalize the surface of the apatite using lauroyl chloride to improve the dispersion of the mineral phase in a poly(L-lactide) (PLLA) polymeric matrix. Infrared spectra and thermal gravimetric analysis confirmed the presence of laurate on the surface of calcium phosphate (CaP) particles. Neat HA particles were also functionalized with lauryl chloride for comparative purposes. PLLA/OCP/HA-laurate (PLLA/OCP/HA-L) and PLLA/HA-laurate (PLLA/HA-L) composites were fabricated by electrospinning method. The presence of the functional groups resulted in significant improvement of the dispersion of OCP/HA and HA particles into the polymeric matrix, allowing inclusion of up to 40% of mineral phase. Electrospun fibrous biocomposites of PLLA/CaP containing up to 40% in mineral phase were obtained without compromising their mechanical properties. Measurements of mass loss and calcium release in vitro showed that OCP/HA is more soluble than HA. The bioactivity of the composites was investigated by simulated body fluid test (SBF). Although both PLLA/OCP/HA-L and PLLA/HA-L fibers can form CaP crystals on their surface after exposition in SBF, the results demonstrate a significant enhancement in mineralization when OCP/HA is the mineral phase in the composite instead neat HA. Furthermore, the obtained PLLA/OCP/HA-L electrospun fibers favored the proliferation and differentiation of stem cells from human exfoliated deciduous teeth and mouse calvaria-derived preosteoblastic cells into mineralized osteoblasts. This new material is proposed as fast degrading CaP biocomposite for bone and teeth applications.

Graphene Oxide-A Tool for the Preparation of Chemically Crosslinking Free Alginate-Chitosan-Collagen Scaffolds for Bone Tissue Engineering.

Developing a biodegradable scaffold remains a major challenge in bone tissue engineering. This study was aimed at developing novel alginate-chitosan-collagen (SA-CS-Col)-based composite scaffolds consisting of graphene oxide (GO) to enrich porous structures, elicited by the freeze-drying technique. To characterize porosity, water absorption, and compressive modulus, GO scaffolds (SA-CS-Col-GO) were prepared with and without Ca2+-mediated crosslinking (chemical crosslinking) and analyzed using Raman, Fourier transform infrared (FTIR), X-ray diffraction (XRD), and scanning electron microscopy techniques. The incorporation of GO into the SA-CS-Col matrix increased both crosslinking density as indicated by the reduction of crystalline peaks in the XRD patterns and polyelectrolyte ion complex as confirmed by FTIR. GO scaffolds showed increased mechanical properties which were further increased for chemically crosslinked scaffolds. All scaffolds exhibited interconnected pores of 10-250 µm range. By increasing the crosslinking density with Ca2+, a decrease in the porosity/swelling ratio was observed. Moreover, the SA-CS-Col-GO scaffold with or without chemical crosslinking was more stable as compared to SA-CS or SA-CS-Col scaffolds when placed in aqueous solution. To perform in vitro biochemical studies, mouse osteoblast cells were grown on various scaffolds and evaluated for cell proliferation by using MTT assay and mineralization and differentiation by alizarin red S staining. These measurements showed a significant increase for cells attached to the SA-CS-Col-GO scaffold compared to SA-CS or SA-CS-Col composites. However, chemical crosslinking of SA-CS-Col-GO showed no effect on the osteogenic ability of osteoblasts. These studies indicate the potential use of GO to prepare free SA-CS-Col scaffolds with preserved porous structure with elongated Col fibrils and that these composites, which are biocompatible and stable in a biological medium, could be used for application in engineering bone tissues.

In vitro analysis of a local polymeric device as an alternative for systemic antibiotics in Dentistry.

The development of a biodegradable material with antimicrobial properties for local applications is required in the prevention and treatment of infectious diseases. The objective of this study was to produce blends of poly-L-lactide acid (PLLA) synthetic polymer associated with several antimicrobials, as an alternative in the prevention and treatment of infections, as well as to evaluate its cytotoxicity, release of antimicrobials and inhibit bacteria growth. Blends of PLLA added with 20% Amoxicillin, Metronidazole, Clindamycin or Azithromicyn were used to produce Films (F) or Meshs (M) by casting and electrospinning methods, respectively. Standardized discs of the films and meshs were stored in buffer solutions (pH 5 or 7.4) and aliquots were analyzed by high performance chromatography (HPLC) during 168 hours. Cytotoxicity on human gingival fibroblasts was tested after 24, 48 and 72h by MTT reaction. The antimicrobial capacity was determined against P. gingivalis and S. pyogenes. The specimens were weighed after 3 and 6 months of storage for degradation analysis. SEM was performed to control interfaces and degradation. Antimicrobials presented a continuous and exponential drug release. Analysis showed that both M and F were able to inhibit S. pyogenes and P. gingivalis growth, indicating the release of active antimicrobial agents. The products were not toxic to the fibroblasts. Amoxicillin-film showed more degradation than PLLA at both pHs (p < 0.05), whereas Azithromycin-meshes were more degraded than PLLA at pH 7.4 (p < 0.05). PLLA association with antimicrobials is biocompatible and may represent a potential tool for the local delivery of antimicrobials.

Hybrid magnetic scaffolds: The role of scaffolds charge on the cell proliferation and Ca2+ ions permeation.

Magnetic scaffolds with different charge densities were prepared using magnetic nanoparticles (MNP) and xanthan gum (XG), a negatively charged polysaccharide, or hydroxypropyl methylcellulose (HPMC), an uncharged cellulose ether. XG chains were crosslinked with citric acid (cit), a triprotic acid, whereas HPMC chains were crosslinked either with cit or with oxalic acid (oxa), a diprotic acid. The scaffolds XG-cit, HPMC-cit and HPMC-oxa were characterized by scanning electron microscopy (SEM), inductively coupled plasma atomic emission spectroscopy (ICP-AES), superconducting quantum interference device (SQUID) magnetometry, contact angle and zeta-potential measurements. In addition, the flux of Ca2+ ions through the scaffolds was monitored by using a potentiometric microsensor. The adhesion and proliferation of murine fibroblasts (NIH/3T3) on XG-cit, XG-cit-MNP, HPMC-cit, HPMC-cit-MNP, HPMC-oxa and HPMC-oxa-MNP were evaluated by MTT assay. The magnetic scaffolds presented low coercivity (<25Oe). The surface energy values determined for all scaffolds were similar, ranging from 43mJm-2 to 46mJm-2. However, the polar component decreased after MNP incorporation and the dispersive component of surface energy increased in average 1mJm-2 after MNP incorporation. The permeation of Ca2+ ions through XG-cit-MNP was significantly higher in comparison with that on XG-cit and HPMC-cit scaffolds, but through HPMC-cit-MNP, HPMC-oxa and HPMC-oxa-MNP scaffolds it was negligible within the timescale of the experiment. The adhesion and proliferation of fibroblasts on the scaffolds followed the trend: XG-cit-MNP>XG-cit>HPMC-cit, HPMC-cit-MNP, HPMC-oxa, HPMC-oxa-MNP. A model was proposed to explain the cell behavior stimulated by the scaffold charge, MNP and Ca2+ ions permeation.

In vitro analysis of a local polymeric device as an alternative for systemic antibiotics in Dentistry

Abstract The development of a biodegradable material with antimicrobial properties for local applications is required in the prevention and treatment of infectious diseases. The objective of this study was to produce blends of poly-L-lactide acid (PLLA) synthetic polymer associated with several antimicrobials, as an alternative in the prevention and treatment of infections, as well as to evaluate its cytotoxicity, release of antimicrobials and inhibit bacteria growth. Blends of PLLA added with 20% Amoxicillin, Metronidazole, Clindamycin or Azithromicyn were used to produce Films (F) or Meshs (M) by casting and electrospinning methods, respectively. Standardized discs of the films and meshs were stored in buffer solutions (pH 5 or 7.4) and aliquots were analyzed by high performance chromatography (HPLC) during 168 hours. Cytotoxicity on human gingival fibroblasts was tested after 24, 48 and 72h by MTT reaction. The antimicrobial capacity was determined against P. gingivalis and S. pyogenes. The specimens were weighed after 3 and 6 months of storage for degradation analysis. SEM was performed to control interfaces and degradation. Antimicrobials presented a continuous and exponential drug release. Analysis showed that both M and F were able to inhibit S. pyogenes and P. gingivalis growth, indicating the release of active antimicrobial agents. The products were not toxic to the fibroblasts. Amoxicillin-film showed more degradation than PLLA at both pHs (p < 0.05), whereas Azithromycin-meshes were more degraded than PLLA at pH 7.4 (p < 0.05). PLLA association with antimicrobials is biocompatible and may represent a potential tool for the local delivery of antimicrobials.

Combination of Bioactive Polymeric Membranes and Stem Cells for Periodontal Regeneration: In Vitro and In Vivo Analyses.

Regeneration of periodontal tissues requires a concerted effort to obtain consistent and predictable results in vivo. The aim of the present study was to test a new family of bioactive polymeric membranes in combination with stem cell therapy for periodontal regeneration. In particular, the novel polyester poly(isosorbide succinate-co-L-lactide) (PisPLLA) was compared with poly(L-lactide) (PLLA). Both polymers were combined with collagen (COL), hydroxyapatite (HA) and the growth factor bone morphogenetic protein-7 (BMP7), and their osteoinductive capacity was evaluated via in vitro and in vivo experiments. Membranes composed of PLLA/COL/HA or PisPLLA/COL/HA were able to promote periodontal regeneration and new bone formation in fenestration defects in rat jaws. According to quantitative real-time polymerase chain reaction (qRT-PCR) and Alizarin Red assays, better osteoconductive capacity and increased extracellular mineralization were observed for PLLA/COL/HA, whereas better osteoinductive properties were associated with PisPLLA/COL/HA. We concluded that membranes composed of either PisPLLA/COL/HA or PLLA/COL/HA present promising results in vitro as well as in vivo and that these materials could be potentially applied in periodontal regeneration.

Biocompatible xanthan/polypyrrole scaffolds for tissue engineering.

Polypyrrole (PPy) was electropolymerized in xanthan hydrogels (XCA), resulting in electroactive XCAPPy scaffolds with (15 ± 3) wt.% PPy and (40 ± 10) µm thick. The physicochemical characterization of hybrid XCAPPy scaffolds was performed by means of cyclic voltammetry, swelling tests, Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), thermogravimetric analyses (TGA), scanning electron microscopy (SEM), atomic force microscopy (AFM) and tensile tests. XCAPPy swelled~80% less than XCA. FTIR spectra and thermal analyses did not evidence strong interaction between PPy and XCA matrix. XCAPPy presented a porous stratified structure resulting from the arrangement of PPy chains parallel to XCA surface. Under stress XCAPPy presented larger strain than neat XCA probably due to the sliding of planar PPy chains. The adhesion and proliferation of fibroblasts onto XCA and XCAPPy were evaluated in the absence and in the presence of external magnetic field (EMF) of 0.4T, after one day, 7 days, 14 days and 21 days. Fibroblast proliferation was more pronounced onto XCAPPy than onto XCA, due to its higher hydrophobicity and surface roughness. EMF stimulated cell proliferation onto both scaffolds.

Synthesis and characterization of xanthan-hydroxyapatite nanocomposites for cellular uptake.

In this work xanthan-nanohydroxyapatite (XnHAp) and its equivalent strontium substituted (XnHApSr) were synthesized by the precipitation of nanohydroxyapatite in xanthan aqueous solution, characterized and compared to conventional hydroxyapatite particles (HAp). XnHAp and XnHApSr were less crystalline than HAp, as revealed by X-ray diffraction. Xanthan chains enriched the surface of XnHAp and XnHApSr particles, increasing the zeta potential values from -(7±1)mV, determined for HAp, to -(17±3)mV and -(25±3)mV, respectively. This effect led to high colloidal stability of XnHAp and XnHApSr dispersions and acicular particles (140±10)nm long and (8±2)nm wide, as determined by scanning electron microscopy and atomic force microscopy. XnHAp and XnHApSr particles were added to xanthan hydrogels to produce compatible nanocomposites (XCA/XnHAp and XCA/XnHApSr). Dried nanocomposites presented surface energy, Young's modulus and stress at break values comparable to those determined for bare xanthan matrix. Moreover, adding XnHAp or XnHApSr nanoparticles to xanthan hydrogel did not influence its porous morphology, gel content and swelling ratio. XCA/XnHAp and XCA/XnHApSr composites proved to be suitable for osteoblast growth and particularly XCA/XnHapSr composites induced higher alkaline phosphatase activity.

Cytotoxicity of PVPAC-treated bovine pericardium: a potential replacement for glutaraldehyde in biological heart valves.

Acellular biological tissues, including bovine pericardium (BP), have been proposed as biomaterial for tissue engineering. BP is usually modified chemically to improve mechanical and biological properties using glutaraldehyde, the standard reagent for preservation of fresh bioprosthetic materials. Glutaraldehyde-fixed BP (Glut-BP), the most widely used material in heart valve manufacture, has been associated with calcification in vivo. In an attempt to reduce this issue and maintain its biocompatibility, this study assesses the physical properties and cytotoxicity of lyophilized BP treated with poly (vinylpyrrolidone-co-acrolein) (PVPAC-BP), a novel copolymer, as a substitute for glutaraldehyde. For that, PVPAC-BP surface ultrastructure, elastic function, water uptake and tissue calcification were evaluated. For the analysis of biocompatibility, fibroblasts (3T3-L1) and endothelial cells (HUVEC) were cultured on PVPAC-BP, Untreated-BP and Glut-BP. Nitric oxide (NO) release assay, fluorescence and SEM images of endothelial cells adhered on scaffolds were also performed. As results, the data show some advantages of PVPAC-BP over the Glut-BP. The PVPAC-BP maintains partially the original ultrastructure and elastic properties, improves scaffold hydration, and presents less calcium phosphate deposits. The cells demonstrated strong attachment, high proliferation rate, and formation of a monolayer on PVPAC-BP. Attached cells were also able to release NO de-monstrating regular metabolism. In conclusion, PVPAC may be considered as a promising alternative to BP treatment improving the efficiency of cell attachment and proliferation and also avoid immunogenicity.

Hybrid composites of xanthan and magnetic nanoparticles for cellular uptake.

We describe a fast and simple method to prepare composite films of magnetite nanoparticles and xanthan networks. The particles are distributed close to hybrid film surface, generating a coercivity of 27 ± 2 Oe at 300 K. The proliferation of fibroblast cells on the hybrid composites was successful, particularly when an external magnetic field was applied.