Alginate templated synthesis, characterization and in vitro osteogenic evaluation of strontium-substituted hydroxyapatite.

The objective of this study is to explore the potential role of alginate (Alg) in the crystallization of metal-substituted hydroxyapatite, with application in orthopaedic reconstruction. The alginate at different concentrations (0.5 and 1.0 wt%) facilitated in situ mineralization of hydroxyapatite (HA) and strontium-substituted HA (SHA, 10 and 30 mol%). The incorporation of the biopolymer and dopant induced notable changes in HA, including reduced crystal size from 31.0 to 16.4 nm and increased lattice volume from 577.3 to 598.0 Å3. The superior affinity of alginate for Sr2+ than for Ca2+ resulted in higher residual alginate in Alg/SHA (13.0 to 19.0 %) compared to Alg/HA (7.1 to 8.2 %). This residual alginate influenced composite properties: surface charge decreased from -26.5 to -45.7 mV, microhardness increased from 0.33 to 0.54 GPa, and dissolution increased from 0.17 to 0.39 %. The in vitro studies revealed that strontium substitution as well as the organization and crystallographic aspects of apatite regulated osteoblastic cell survival, proliferation, differentiation, and biomineralization. The findings suggest that an alginate concentration of 0.5 wt% is optimal for the crystallization of SHA with 10 mol% substitution, and its resulting composite possesses the ideal biomechanical properties to imitate native bone.

Dextrose, maltose and starch guide crystallization of strontium-substituted hydroxyapatite: A comparative study for bone tissue engineering application.

The influence of carbohydrates on the crystallization of metal-substituted hydroxyapatite predicts its relevance to natural bone growth. This study demonstrates the role of carbohydrates in the crystallization of strontium-substituted hydroxyapatite (SHAP). The increasing order of hydroxyl groups, dextrose (monosaccharide) < maltose (disaccharide) < starch (polysaccharide), coordinated with Ca2+/Sr2+ and thus guided SHAP crystallization, with crystal size reduced from 35 to 19 nm, lattice volume increased from 518 to 537 Å3, and residual carbohydrates increased from 1.8 to 20.2 %. The variation in residual carbohydrates is due to their interaction with apatite and/or aqueous insolubility. Compared to pure SHAP, the starch-SHAP with higher residual starch showed increased water uptake from 1.23 ± 0.18 to 4.26 ± 0.21 % and degradation from 0.22 ± 0.06 to 1.53 ± 0.14 %, but decreased microhardness from 0.73 ± 0.12 to 0.38 ± 0.01 GPa and protein affinity from 4.82 ± 0.01 to 0.81 ± 0.01 µg/mg. However, its microhardness value was bone-like, and the reduced protein adsorption was masked by the rich osteogenic behaviour. In vitro cellular response demonstrated that the residual carbohydrate and strontium augmented osteocompatibility, proliferation, differentiation and biomineralization. The result concludes that carbohydrates drive SHAP crystallization, and starch-SHAP replicates natural bone.

Effect of pectin on the crystallization of strontium substituted HA for bone reconstruction application.

The biomacropolymers of bone extracellular matrix (ECM) guide the growth of hydroxyapatite (HA) with various ionic substitutions. Pectin, a plant polysaccharide with chemical similarities to ECM, was investigated for its potential to promote the crystallization of strontium-substituted HA (SH). The influence of pectin (0.5 and 1.0 wt%) on the in situ mineralization of SH (10 and 30 mol% calcium substitution with strontium) was studied. The preferential affinity of pectin to strontium over calcium favoured the incorporation of strontium in apatite, decreased crystal size (18.85-26.22 nm) and retained more pectin residues (8-16%). The residual pectin strongly interacted with small SH particles, resulting in high microhardness (0.43-0.85 GPa) and high surface charge (-32.1 to -30.3 mV), while weak interaction with large HA particles resulted in low microhardness (0.15-0.25 GPa) and low surface charge (-35.4 to -34.6 mV). The in vitro cellular study using human osteoblast-like MG-63 cells demonstrated that inorganic size and material crystallinity play a vital role in regulating osteogenesis. The study suggests that the synchronization of low pectin concentration (0.5 wt%) and high strontium substitution in HA (30 mol%) offers the desired microhardness and in vitro osteogenic properties to emulate natural bone.

Graphene oxide reinforced SrHAP composite as a drug carrier in bone regeneration.

Strontium substituted HAP (SrHAP), with a 10 mol% substitution, was mineralized on increasing weight percentages of graphene oxide (2, 4 and 6). The GS composites were comprehensively characterized for drug delivery in bone reconstruction. The formation of SrHAP was verified by XRD and FT-IR results. The apatite crystallization was influenced by graphene oxide content and strontium. The EDS results confirmed the presence of strontium and HR-SEM depicted rod shape apatite, of length between 58 and 135 nm, uniformly embedded on graphene oxide. The reinforcement of graphene oxide increased the surface area, porosity, microhardness (upto 0.59 GPa), protein adsorption (upto 18.16 µg/mg), water uptake and degradation properties. Also, the increase in graphene oxide fraction significantly enhanced the curcumin encapsulation efficiency (upto 80.16%) and the drug release was considerably retarded over SrHAP. The in vitro studies using human osteoblast-like MG-63 cells demonstrated that curcumin-loaded composite was biocompatible and promoted proliferation, differentiation and matrix mineralization. The results highlight the combinational therapy of osteogenic ion (strontium) and osteogenic drug (curcumin) as a promising platform in bone tissue engineering.

Evaluation of quaternization effect on chitosan-HAP composite for bone tissue engineering application.

This study attempts to improve the aqueous solubility of chitosan and utilizes it in the fabrication of composites with hydroxyapatite (HAP). The composites were evaluated as a curcumin delivery vehicle for bone regeneration. The chitosan was modified by quaternization, with a quaternization degree of 5 % for low quaternized chitosan (LQC) and 11 % for high quaternized chitosan (HQC). The modified chitosan, at alkaline pH 11, facilitated in situ HAP growth and formed LQC-HAP and HQC-HAP composites. The quaternization weakens intermolecular hydrogen bonds, facilitates interaction with the apatite precursor ions and promotes the growth of HAP. The modification significantly improved drug encapsulation (2.6 fold) but at the cost of a slight decrease in mechanical strength and increase in drug release. The in vitro studies with human osteoblast-like MG-63 cells established that the curcumin-loaded composites, LQC-HAP-C and HQC-HAP-C are biocompatible, encourage proliferation and promote a 2-fold increase in calcium mineralization over drug-free composites. The study exemplifies the reciprocity between quaternization degree and drug load/release properties and also illustrates that the magnitude of the latter reflects bioactivity. Thus, the quaternized chitosan-based HAP composite with tailorable bio-physicochemical properties becomes an interesting drug delivery system in bone regeneration.

Multiwalled Carbon Nanotubes/Gold Nanoparticles Hybrid Electrodes for Enzyme-Free Electrochemical Glucose Sensor.

We followed a facile strategy to fabricate glucose sensors using mildly oxidized multiwalled carbon nanotubes (MWCNTs), gold nanoparticles (AuNPs) and thiol acids including mercaptoacetic acid (MAA), mercaptopropionic acid (MPA), and mercaptosuccinic acid (MSA). The thiol acids separately bonded to MWCNTs anchored AuNPs of average diameter 14 nm, and yielded three different nanohybrids; MWCNTs-MAA-AuNPs, MWCNTs-MPA-AuNPs and MWCNTs-MSA-AuNPs. The nanohybrids after coating onto glassy carbon (GC) electrode resulted into enzyme free glucose sensors (GC-MWCNTs-MAA-AuNPs, GC-MWCNTs-MPA-AuNPs and GC-MWCNTs-MSA-AuNPs). Their electrocatalytic glucose sensing ability was examined through cyclic voltammetry and amperometry. GC-MWCNTs-MSA-AuNPs electrode showed high stability and activity in the electrocatalytic oxidation of glucose compared to other two sensors. It also showed a wide range of response for glucose concentration from 0.12 to 4.0 µM, and low detection limit of 0.036 µM (S/N = 3). The optimum rate of applied potential was 0.8 V/s, which proves the effective sensing of glucose. The selective sensing of glucose in the presence of H2O2, uric acid and blood cancer drug (imatinib mesylate) was verified through amperometry. The electrode can be a new addition to glucose sensors and bioanalytical techniques.

Core-Shell Nanostructured Fe3O4-Poly(styrene-co-vinylbenzyl chloride) Grafted PPI Dendrimers Stabilized with AuNPs/PdNPs for Efficient Nuclease Activity.

Four different novel magnetic core-shell nanocomposites stabilized with Au/Pd nanoparticles (NPs) were prepared by a simple procedure and demonstrated their catalytic activity for effective cleavage of pBR322 DNA. Initially, the Fe3O4-poly(styrene-divinylbenzene-vinylbenzyl chloride) (ST-DVB-VBC) matrix functionalized with 3-aminobenzoic acid was prepared and grafted with PPI-G(2) and PPI-G(3) dendrimers. Each core-shell matrix was immobilized with AuNPs and PdNPs separately. The resulting composites were characterized by FT-IR, UV-vis, SEM, TEM, XRD, VSM, XPS, Raman, and TGA. The magnetic core-shell nanocomposites at concentrations from 30 to 50 µM were employed separately to study DNA cleavage by agarose gel electrophoresis. Among the four magnetic core-shell nanocomposites, Fe3O4-poly(ST-DVB-VBC)-PPI-G(3)-AuNPs showed higher activity than others for DNA cleavage, and formed Form-II and -III DNA. When the concentration of Fe3O4-poly(ST-DVB-VBC)-PPI-G(3)-AuNPs was increased from 40 to 45 and 45 to 50 µM, Form-III (linear) DNA was observed with 10 and 22%, respectively, in addition to Form-II. This observation suggests formation of linear DNA from the supercoiled DNA via nicked DNA-intermediated consecutive cleaving process. The magnetic core-shell nanocomposites were stable and monodispersed, and exhibited rapid magnetic response. These properties are crucial for their application in biomolecular separations and targeted drug-delivery in the future.