Probing the influence of strontium doping and annealing temperature on the structure and biocompatibility of hydroxyapatite nanorods.

Among numerous biologically important metal cations, strontium (Sr2+) has received much attention in bone tissue regeneration because of its osteoinductive properties combined with its ability to inhibit osteoclast activity. In this study, strontium-doped hydroxyapatite (Sr-HAp) nanorods with varying molar ratios of Ca : Sr (10 : 0, 9 : 1, 5 : 5, 3 : 7 and 0 : 10) were synthesized using the chemical precipitation technique. The synthesized Sr-HAp nanostructures were characterized using powder X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), transmission electron microscopy, energy dispersive X-ray spectroscopy, and Raman and Fourier transform infrared (FTIR) spectroscopies to understand their structural and morphological features, and composition. XRD results revealed the formation of HAp nanostructures, whose unit cell volume increased as a function of the dopant level. The reaction process investigation showed the formation of hydroxyapatite (HAp), strontium apatite (SAp) and various Sr-HAp phases. FESEM micrographs displayed the morphological transformation of Sr-HAp from nanorods to nanosheets upon increasing the dopant level. In the FTIR spectra, the bands of the PO43- group shifted towards a lower wavenumber upon increasing the dopant concentration in Sr-HAp that signifies the structural distortion due to the presence of a large amount of strontium ions. The peaks of PO43- and OH- vibrations in the Raman spectra were further analysed to corroborate the structural distortion of Sr-HAp. Selected area electron diffraction patterns obtained using TEM reveal the reduced crystallinity of Sr-HAp due to Sr-doping, which is in line with the XRD results. Finally, the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay showed that the synthesized Sr-HAp has no toxic effect on the survival and growth of mesenchymal stem cells. In summary, the synthesized novel Sr-HAp nanorods exhibit great promise for bone tissue engineering applications.

Investigation of human hair keratin-based nanofibrous scaffold for skin tissue engineering application.

Keratin-based nanofibers were fabricated using the electrospinning technique, and their potential as scaffolds for tissue engineering was investigated. Keratin, extracted from the human hair, was blended with poly(vinyl alcohol) (PVA) in an aqueous medium. Morphological characterizations of the fabricated PVA-keratin nanofiber (PK-NF) random and aligned scaffolds performed using a scanning electron microscope (SEM) revealed the formation of uniform and randomly oriented nanofibers with an interconnected three-dimensional network structure. The mean diameter of the nanofibers ranged from 100 to 250 nm. Functional groups and structural studies were done by infrared spectroscopy (FTIR) and X-ray diffraction (XRD) analysis. FTIR study suggested that PVA interacted with keratin by hydrogen bonding. Moreover, the in vitro cell culture study could suggest that PK-NF scaffolds were non-cytotoxic by supporting the growth of murine embryonic stem cells (ESCs), human keratinocytes (HaCaT), and dermal fibroblast (NHDF) cell lines. Further, the immunocytochemical characterization revealed the successful infiltration, adhesion, and growth of ESCs, HaCaT, and NHDF cells seeded on PK-NF scaffolds. However, there was no noteworthy difference observed concerning cell growth and viability irrespective of the random and aligned internal fibril arrangement of the PK-NF scaffolds. The infiltration and growth pattern of HaCaT and NHDF cells adjacent to each other in a 3D co-culture study mimicked that of epidermal and dermal skin cells and indeed underscored the potential of PK-NFs as a scaffold for skin tissue engineering.

Development of bioactive and antimicrobial nano-topography over selective laser melted Ti6Al4V implant and its in-vitro corrosion behavior.

Additive manufacturing (AM) or 3D printing of bone defect models is gaining much attention in the biomedical field as it could significantly facilitate the development of customized implants with a high degree of dimensional accuracy. Due to their satisfactory biocompatibility and minimal stress shielding effect, Ti6Al4V (Ti64) alloys are increasingly preferred in the development of such implants. However, their poor osseointegration abilities and lack of antibacterial properties often cause implant loosening and microbial infections, leading to implant failure. To address these drawbacks, we propose in this work a simple surface modification approach of customized Ti64 alloys (3D printed Ti6Al4V) that enables the formation of porous calcium titanate (CT) over their surface as well as the incorporation of silver nanoparticles (AgNPs) into the thus formed porous network. The successful CT formation with the incorporation of AgNPs throughout the 3D printed Ti64 surface and their influence in changing the morphological and mechanical behaviour were studied by Raman spectroscopy, SEM, AFM, Contact angle measurement, XPS, HR-TEM and nano-indentation. Antibacterial studies using Staphylococcus aureus and Escherichia coli, and in-vitro cell studies using MG-63 cell lines showed that surface modified samples resulting from the proposed method exhibit satisfactory antimicrobial property and are highly biocompatible. The obtained surface modified samples also showed a significant improvement in corrosion resistance as compared to unmodified 3D printed Ti64 alloys. The improvement in corrosion resistance was revealed by electrochemical impedance Spectroscopy (EIS). Obtained results emphasis that thus surface modified 3D printed Ti64 alloys are promising candidates for hard tissue implant applications.

Fabrication and characterization of ceramic-polymer composite 3D scaffolds and demonstration of osteoinductive propensity with gingival mesenchymal stem cells.

The fabrication of biomaterial 3D scaffolds for bone tissue engineering applications involves the usage of metals, polymers, and ceramics as the base constituents. Notwithstanding, the composite materials facilitating enhanced osteogenic differentiation/regeneration are endorsed as the ideally suited bone grafts for addressing critical-sized bone defects. Here, we report the successful fabrication of 3D composite scaffolds mimicking the ECM of bone tissue by using ∼30 wt% of collagen type I (Col-I) and ∼70 wt% of different crystalline phases of calcium phosphate (CP) nanomaterials [hydroxyapatite (HAp), beta-tricalcium phosphate (ßTCP), biphasic hydroxyapatite (ßTCP-HAp or BCP)], where pH served as the sole variable for obtaining these CP phases. The different Ca/P ratio and CP nanomaterials orientation in these CP/Col-I composite scaffolds not only altered the microstructure, surface area, porosity with randomly oriented interconnected pores (80-450 µm) and mechanical strength similar to trabecular bone but also consecutively influenced the bioactivity, biocompatibility, and osteogenic differentiation potential of gingival-derived mesenchymal stem cells (gMSCs). In fact, BCP/Col-I, as determined from micro-CT analysis, achieved the highest surface area (∼42.6 m2 g-1) and porosity (∼85%), demonstrated improved bioactivity and biocompatibility and promoted maximum osteogenic differentiation of gMSCs among the three. Interestingly, the released Ca2+ ions, as low as 3 mM, from these scaffolds could also facilitate the osteogenic differentiation of gMSCs without even subjecting them to osteoinduction, thereby attesting these CP/Col-I 3D scaffolds as ideally suited bone graft materials.

Bioactive and antimicrobial macro-/micro-nanoporous selective laser melted Ti-6Al-4V alloy for biomedical applications.

Metal Additive Manufacturing (AM) technology is an emerging technology in biomedical field due to its unique ability to manufacture customized implants [Patients-specific Implants (PSIs)] replicating the complex bone structure from the relevant metal powders. PSIs could be developed through any AM technology, but the ultimate challenge lies in integrating the metallic implant with the living bone. Considering this aspect, in the present study, Ti alloy (Ti-6Al-4V) powder has been used to fabricate scaffolds of channel type macropores with 0-60% porosity using selective laser melting (SLM) and subsequent post-treatments paving way for surface microporosities. Surface chemical and subsequent heat treatments were carried out on thus developed Ti alloy scaffolds to improve its bioactivity, antibacterial activity and osteoblastic cell compatibility. NaOH and subsequent Ca(NO3)2/AgNO3 treatment induced the formation of a nanoporous network structure decorated with Ca-Ag ions. Ag nanoparticles covering the entire scaffold provided antibacterial activity and the presence of Ca2+ ions with anatase TiO2 layer further improved the bioactivity and osteoblastic cell compatibility of the scaffold. Therefore, SLM technology combined with heat treatment and surface modification could be effectively utilized to create macro-micro-nano structure scaffolds of Ti alloy that are bioactive, antibacterial, and cytocompatible.

Effects of Carboxymethyl Modification on the Acidic Polysaccharides from Calocybe indica: Physicochemical Properties, Antioxidant, Antitumor and Anticoagulant Activities.

An acidic polysaccharide fraction was obtained from Calocybe indica (CIP3a) after subjecting it to hot water extraction followed by purification through DEAE-cellulose 52 and Sepaharose 6B column chromatography. The CIP3a was further modified using chloroacetic acid to yield carboxymethylated derivatives (CMCIP3a). The modified polysaccharide was characterized using various spectroscopic methods. In addition, further antioxidant, antitumor and anticoagulant activities were also investigated. The polysaccharides CIP3a and CMCIP3a were heterogeneous in nature and composed of various molar percentages of glucose, arabinose and mannose with molecular weights of 1.456 × 103 and 4.023 × 103 Da, respectively. The NMR and FT-IR data demonstrated that the carboxymethylation on the polysaccharide was successful. In comparison to CIP3a polysaccharides, the modified derivatives had lower sugar and protein contents, and higher levels of uronic acid. The in vitro antioxidant activity showed that CMCIP3a with higher molecular weight displayed an elevated ability in scavenging the DPPH radical, ABTS, superoxide, hydroxyl radical, ferric reducing power, cupric reducing power and erythrocyte hemolysis inhibition with an EC50 value of 2.49, 2.66, 4.10, 1.60, 3.48, 1.41 and 2.30 mg/mL, respectively. The MTT assay results revealed that CMCIP3a displayed a dose-dependent inhibition on five cancer cells (HT29, PC3, HeLa, Jurkat and HepG-2) in the range of 10-320 µg/mL. The APTT, PT and TT were significantly extended by CMCIP3a in relation to dosage, indicating that the anticoagulant effect of CIP was both extrinsic and intrinsic, along with a common coagulation pathway. These findings demonstrated that carboxymethylation might effectively improve the biological potential of the derivatives and offer a theoretical framework for the creation of novel natural antioxidants, low-toxicity antitumor and antithrombotic drugs.

Mechanistic studies of biomineralisation on silver incorporated anatase TiO2.

Here we report silver incorporated anatase TiO2 developed on Ti metal by H2O2-AgNO3 and heat treatment to have faster biomineralisation or apatite-forming ability in simulated body fluid (SBF). Apatite-forming ability has been investigated concerning heat treatment temperatures ranges, 400-800 °C and duration of soaking period in SBF. The apatite formation showed an increasing trend with increase in the heat treatment temperatures up to 600 °C and beyond that the Ti metal lost this ability. XRD as wells as Raman results of such chemical and heat-treated Ti metal at different temperatures further correlates the apatite nucleation directly in relation with that of anatase to rutile TiO2 formation. Further, a time dependent apatite mineralisation study by XPS revealed simultaneous calcium and phosphate deposition at the early stage of soaking in SBF. Therefore, the apatite nucleation in the present chemically treated Ti metal depends on the crystalline phase of TiO2 formed by H2O2 and heat treatment along with Ag+ ion release.

Ca-Ag coexisting nano-structured titania layer on Ti metal surface with enhanced bioactivity, antibacterial and cell compatibility.

A nano-structured titanate layer encapsulated with Ca2+ and Ag+ ions was successfully grown over commercially pure (CP) Ti metal by chemical treatment with H2O2 and subsequent treatment with Ca (NO3)2/AgNO3 solutions. Heat treatment at 600 °C, further transformed this nano-structured titanate layer into titania containing Ca2+ and Ag+ ions. Thus modified Ti metal showed significant enhancement in apatite-forming ability when soaked in simulated body fluid (SBF). Presence of Ag+ ions showed good antimicrobial activity against pathogenic Staphylococcus aureus, and, Ca2+ ions being a major component of bone mineral accelerated the apatite-forming ability over Ti metal in SBF. Further, Ca2+and Ag+ ions proportion over Ti metal surface could be optimised in order to have minimum Ag concentration that can have not only antibacterial activity and also cell compatibility against MG 63 osteoblast-like cells. Therefore, the proposed surface modification approach presented here is expected to be useful in orthopaedic implants that necessitate enhanced bioactivity, antibacterial activity and cell compatibility.

Role of calcium ions in defining the bioactivity of surface modified Ti metal.

Nano-structured hydrogen titanate and sodium hydrogen titanate layers were formed when Ti metal was treated with H2O2 and NaOH solutions, respectively. The chemically treated Ti metals upon subsequent treatment with Ca(NO3)2 and CaCl2 solutions, resulted in incorporation of Ca2+ ions into the nano-structured titanate layer. Thus formed nano-structured titanate layers containing Ca2+ ions when subjected to heat treatment, forms anatase and calcium titanate-rutile phases, respectively. In vitro apatite-forming ability in simulated body fluid (SBF) was positive for H2O2-Ca and heat-treated Ti metal in contrast to NaOH-Ca and heat treatment. Formation of anatase phase together with Ca2+ ion release into SBF was found to be the key driving force for such a high bioactivity of Ca2+ containing H2O2 treated Ti metal on contrary to NaOH and heat treatment. This study provides a new insight into the factors accelerating the bioactivity of Ti metals during various chemical and thermal treatments, which further aid and abet to design dental and orthopaedic implants with high bone-bonding ability.

Effect of Silver-Containing Titania Layers for Bioactivity, Antibacterial Activity, and Osteogenic Differentiation of Human Mesenchymal Stem Cells on Ti Metal.

Biomaterials with better osteogenic capacity, rapid osteo-integration, and higher mechanical strength are undoubtedly preferred for successful bone implant development. A porous sodium hydrogen titanate layer was formed on Ti metal by NaOH treatment, and the Na+ ions were replaced by Ag+ ions by subsequent AgNO3 treatment that formed silver-containing hydrogen titanate. Heat treatment at 600 °C transformed sodium hydrogen titanate into sodium titanate with sheet-like morphology, whereas silver-containing hydrogen titanate was converted to anatase TiO2 with an elongated rod-like structure. Further increment in temperature lead to the formation of rutile TiO2 with distracted network morphology. Between these two, the anatase TiO2 was ascertained to be bioactive by being capable of forming bonelike apatite in simulated body fluid within a period of 12 h. The concentration of silver on Ti metal was further optimized for better antibacterial activity against S. aureus and biocompatibility toward bone cells. A detailed investigation of thus optimized silver-containing Ti metal on the proliferation and differentiation of multipotent human mesenchymal stem cells further proved their biocompatibility nature and facilitation of osteogenic differentiation, thereby conferring those as ideally suited materials for bioimplant development in bone tissue engineering.

Comparative study on Ti-Nb binary alloys fabricated through spark plasma sintering and conventional P/M routes for biomedical application.

The main purpose of this work is to obtain homogenous, single ß phase in binary Ti-xNb (x = 18.75, 25, and 31.25 at.%) alloys by simple mixing of pure elemental powders using different sintering techniques such as spark plasma sintering (pressure-assisted sintering) and conventional powder metallurgy (pressure-less sintering). Synthesis parameters such as sintering temperature and holding time etc. are optimized in both techniques in order to get homogenous microstructure. In spark plasma sintering (SPS), complete homogeneous ß phase is achieved in Ti25at.%Nb using 1300 °C sintering temperature with 60 min holding time under 50 MPa pressure. On the other hand, complete ß phase is obtained in Ti25at.%Nb through conventional powder metallurgy (P/M) route using sintering temperature of 1400 °C for 120 min holding time which are adopted from the dilatometry studies. Nano-indentation is carried out for mechanical properties such as Young's modulus and nano-hardness. Elastic properties of binary Ti-xNb compositions are fallen within the range of 80-90 GPa. Cytotoxicity as well as cell adhesion studies carried out using MG63, osteoblast-like cells showed excellent biocompatibility of thus developed Ti25at.%Nb surface irrespective of fabrication route.

Influence of pH on wet-synthesis of silver decorated hydroxyapatite nanopowder.

Here, effect of solution pH on precipitation of silver decorated hydroxyapatite (Ag-HAp) nano powder during its wet-synthesis was systematically studied. XRD pattern of Ag-HAp nano powder synthesised at pH ranging from 5 to 10 shows that calcium hydrogen phosphate was formed as dominating phase when the solution pH was between 5 and 9 and this phase was gradually transformed into a stable HAp above pH 9. A quantitative analysis of silver amount in Ag-HAp nano powder synthesised at different pH showed that silver can be precipitated to its maximum amount at pH 8 and the further addition of ammonia leads to the formation of a silver-ammonium complex, thereby remaining in the solution. HR-TEM and XPS analysis further confirmed the presence of silver in HAp nanocrystals, synthesised at an optimum pH 9. This trace amount of silver in HAp nano powder showed effective antibacterial activity against Staphylococcus aureus and Escherichia coli. In addition, the cytocompatibility studies carried out on MG63 cells further confirmed the present optimised silver concentration of the Ag-HAp nano powder is well within the toxic limit to be useful in various biomedical applications.

Divalent ion encapsulated nano titania on Ti metal as a bioactive surface with enhanced protein adsorption.

A novel approach on incorporation of divalent species such as Mg, Ca and Sr into the titania nanostructures formed on Ti metal surface and their comparative study on enhancement of bioactivity, protein adsorption and cell compatibility is reported. When treated with hydrogen peroxide, Ti metal forms hydrogen titanate. On subsequent treatment with Mg or Ca or Sr nitrate solutions, respective ions are incorporated into hydrogen titanate layer, and heat treatment leads to titania decorated with these ions. The resultant heat-treated samples when soaked in simulated body fluid form bone-like apatite which indicates the present surface modification enhances the bioactivity. Further, enhanced protein adsorption in bovine serum albumin is an indication of suitability of these divalent species to form chelate compounds with amino acids, and Ca containing titania nanostructure favours more protein adsorption compared to the others. Cytocompatibility studies using MG-63, human osteosarcoma cell lines shows these divalent ion containing titania nanostructure favours the cell attachment and did not show any cytotoxicity. Bioactivity, enhanced protein adsorption along with cytocompatibility clearly indicates such surface modification approach to be useful to design hard tissue replacement materials in orthopaedic and dental field.

Predicting properties of organic optoelectronic materials: asymptotically corrected density functional study.

A practical computational procedure has been proposed that provides key electronic parameters of a polymer (fundamental bandgap, ionization energy, electron affinity, and intrachain electron and hole mobilities) determining its suitability as a donor or acceptor in organic optoelectronic materials. Series of oligomer calculations at the Becke3-Lee-Yang-Parr level with and without a self-contained asymptotic correction using the 6-31G** basis set were performed. The bandgap, ionization energy, and electron affinities of a polymer are extrapolated from those of its oligomers obtained from the highest occupied and lowest unoccupied orbital energies in the Koopmans-like approximation. This scheme has been applied to conjugated polymers having the poly(p-phenylene), poly(thiophene), or poly(pyrrole) backbone as well as to PCBM. The observed values of the electronic parameters have been reproduced within less than 1 eV in most cases. With the predicted values of these parameters, estimates of the open-circuit voltage and drift potential have been made for 22 valid donor-acceptor combinations. Several potentially useful combinations have been identified including the poly(thiophene):PCBM. The electron and hole mobilities have been found to correlate more strongly with the conformation (planarity) than the bandgap, but otherwise do not differ significantly.