Biomimetic approaches with smart interfaces for bone regeneration.

A 'smart tissue interface' is a host tissue-biomaterial interface capable of triggering favourable biochemical events inspired by stimuli responsive mechanisms. In other words, biomaterial surface is instrumental in dictating the interface functionality. This review aims to investigate the fundamental and favourable requirements of a 'smart tissue interface' that can positively influence the degree of healing and promote bone tissue regeneration. A biomaterial surface when interacts synergistically with the dynamic extracellular matrix, the healing process become accelerated through development of a smart interface. The interface functionality relies equally on bound functional groups and conjugated molecules belonging to the biomaterial and the biological milieu it interacts with. The essential conditions for such a special biomimetic environment are discussed. We highlight the impending prospects of smart interfaces and trying to relate the design approaches as well as critical factors that determine species-specific functionality with special reference to bone tissue regeneration.

Lawsonia inermis-organically modified chitosan intercalated bentonite clay: A multifunctional nanotheranostic system for controlled drug delivery, sensing and cellular imaging.

This study presents the development of organo-bentonites (OBs); a cost-effective drug delivery system holding both sensing and imaging capabilities. The OBs were synthesized using quaternary ammonium cations derived from chitosan, Lawsonia inermis, and pyrene/anthracene carboxaldehyde combinations through a three-step process: Mannich reaction, quaternization, and intercalation. Physicochemical characterization confirms the organic modification of bentonite. The OBs: NQPB and NQAB hold substantial ciprofloxacin (Cipro) loading capacities (71.51 % and 78.04 %, respectively) and exhibit pH-dependent release profiles, suggesting their potential use as drug delivery platforms. Cell viability evaluation by MTT and live-dead assays indicates favourable results. Both OBs demonstrate fluorescence within the 450-500 nm range, and they display concentration-dependent fluorescence quenching and enhancement for NQPB and NQAB, respectively, in the presence of tryptophan (Trp), making them suitable for its detection. Confocal analysis further enunciates the live intracellular fluorescence upon OB uptake. In summary, the intrinsically fluorescent mesoporous OBs synthesized from Lawsonia inermis and chitosan exhibit multifunctionality, including Cipro delivery, Trp sensing, and live cell imaging. Among the OBs, NQAB could be considered as a promising theranostic platform owing to its superior cytocompatibility (>80 %), appreciable fluorescence, and controlled release profile.

Notch signaling regulates tumor-induced angiogenesis in SPARC-overexpressed neuroblastoma.

Despite existing aggressive treatment modalities, the prognosis for advanced stage neuroblastoma remains poor with significant long-term illness in disease survivors. Advance stage disease features are associated with tumor vascularity, and as such, angiogenesis inhibitors may prove useful along with current therapies. The matricellular protein, secreted protein acidic and rich in cysteine (SPARC), is known to inhibit proliferation and migration of endothelial cells stimulated by growth factors. Here, we sought to determine the effect of SPARC on neuroblastoma tumor cell-induced angiogenesis and to decipher the molecular mechanisms involved in angiogenesis inhibition. Conditioned medium from SPARC-overexpressed neuroblastoma cells (pSPARC-CM) inhibited endothelial tube formation, cell proliferation, induced programmed cell death and suppressed expression of pro-angiogenic molecules such as VEGF, FGF, PDGF, and MMP-9 in endothelial cells. Further analyses revealed that pSPARC-CM-suppressed expression of growth factors was mediated by inhibition of the Notch signaling pathway, and cells cultured on conditioned medium from tumor cells that overexpress both Notch intracellular domain (NICD-CM) and SPARC resumed the pSPARC-CM-suppressed capillary tube formation and growth factor expression in vitro. Further, SPARC overexpression in neuroblastoma cells inhibited neo-vascularization in vivo in a mouse dorsal air sac model. Furthermore, SPARC overexpression-induced endothelial cell death was observed by co-localization studies with TUNEL assay and an endothelial marker, CD31, in xenograft tumor sections from SPARC-overexpressed mice. Our data collectively suggest that SPARC overexpression induces endothelial cell apoptosis and inhibits angiogenesis both in vitro and in vivo.

Multifunctional chitosan ferrogels for targeted cancer therapy by on-demand magnetically triggered drug delivery and hyperthermia.

A facile method for the synthesis of chitosan ferrogels for magnetically triggered drug release and hyperthermia treatment is presented. The glyoxal crosslinked, dried ferrogels (magnetic bioaerogels) have been characterized by FTIR, XRD, TGA and VSM analyses and they possess unique characteristics such as high porosity, ultra-low density and superparamagnetism (Ms up to 56 emu g-1). In addition, they present high drug (Doxorubicin, DOX) loading efficiency (~40 %), tumor-specific pH-responsive swelling, excellent biodegradation, remotely switchable drug release and high magnetic hyperthermia potential (42 °C within 4 min). Almost complete degradation of the ferrogels occurs in 3 months under physiological conditions (pH = 7.4), while the tumor-specific microenvironment (pH = 5.6) accelerates the degradation rate, where it occurs in ~8 weeks. Furthermore, an enhancement in drug release (by 30 %) was observed in 60 min, when subjected to a magnetic field of 50 mT. Excellent biocompatibility and promising cell-material interactions have been exhibited by the ferrogels, substantiated by MTT assay, cytoskeleton staining and confocal imaging. The viability has been drastically reduced for DOX-loaded samples due to the action of the released drug; validating the efficacy of DOX loaded ferrogels. The system presented, therefore, holds multi-functionalities enabling smart cancer treatment.

Radiopacity endowed magnetic nanocomposite with hyperthermia andin vitromineralization potential: a combinatorial therapeutic system for osteosarcoma.

The development of clinically advanced multifaceted therapeutic materials for osteosarcoma is at the forefront of cancer research. Accordingly, this work presents the design of a multifunctional magnetic nanocomposite composed of maghemite, strontium doped hydroxyapatite and silica nanoparticles prospectively holding indispensable therapeutic features such as magnetic hyperthermia,in vitrobiomineralization, sustained drug release and intrinsic radiopacity for the treatment of osteosarcoma. The optimal composition has been identified by sequentially modulating the ratio of precursors of the magnetic nanocomposite synthesized by sol-gel technique. Structural and morphological characterization by x-ray diffraction, fourier transform infrared spectrum, Brunauer-Emmet-Teller and transmission electron microscopy analyses followed by VSM, hyperthermia and micro-CT analyses essentially assisted in the selective configuration of biofunctional properties. Results exemplify that MSHSr1 has a saturation magnetization of 47.4 emu g-1and attained hyperthermia temperature (42 °C) at a very low exposure time of 4 min. MSHSr1 is further unique with respect to its exceptional x-ray attenuation ability (contrast enhancement 154.5% in digital radiography; CT number 3100 HU), early biomimetic mineralization (in vitro) evident by the formation of spheroidal apatite layer (Ca/P ratio 1.33) harvested from FESEM-EDX analysis and controlled release of Doxorubicin, the clinically used chemotherapeutic drug: 87.7% at 120 h in tumour analogous pH (6.5) when compared to physiological pH (71.3% at 7.4). MTT assay complemented with cytoskeleton (F-actin) staining of human osteosarcoma (HOS) cells affirm biocompatibility of MSHSr1.In vitrobiomineralization authenticated by Alizarin red S and von Kossa staining has been further corroborated by semi-quantitative calcium estimation of HOS cells cultured with MSHSr1 for two weeks. The results therefore validate the multifunctionality of MSHSr1, and hence could be proposed as a combinatorial therapeutic nanocomposite for osteosarcoma treatment.

Intrinsically radiopaque biomaterial assortments: a short review on the physical principles, X-ray imageability, and state-of-the-art developments.

X-ray attenuation ability, otherwise known as radiopacity of a material, could be indisputably tagged as the central and decisive parameter that produces contrast in an X-ray image. Radiopaque biomaterials are vital in the healthcare sector that helps clinicians to track them unambiguously during pre and post interventional radiological procedures. Medical imaging is one of the most powerful resources in the diagnostic sector that aids improved treatment outcomes for patients. Intrinsically radiopaque biomaterials enable themselves for visual targeting/positioning as well as to monitor their fate and further provide the radiologists with critical insights about the surgical site. Moreover, the emergence of advanced real-time imaging modalities is a boon to the contemporary healthcare systems that allow to perform minimally invasive surgical procedures and thereby reduce the healthcare costs and minimize patient trauma. X-ray based imaging is one such technologically upgraded diagnostic tool with many variants like digital X-ray, computed tomography, digital subtraction angiography, and fluoroscopy. In light of these facts, this review is aimed to briefly consolidate the physical principles of X-ray attenuation by a radiopaque material, measurement of radiopacity, classification of radiopaque biomaterials, and their recent advanced applications.

In vitro biomineralization and osteogenesis of Cissus quadrangularis stem extracts: An osteogenic regulator for bone tissue engineering.

Contemporary demand calls for a high restorative index as an indispensable requirement for bone tissue engineering scaffolds, where therapeutic agents of natural origin function as a modulator for new bone formation become of utmost importance. The study presents a systematic investigation of the edible stem part of Cissus quadrangularis (CQ) as a natural resource of bioactive metabolites capable of invoking early biomineralization and osteogenesis in vitro. Phytochemical screening of CQ stem extracts (sequential solvent extraction: polarity hexane

Injectable polyelectrolyte complex-nascent HAP biodegradable antibiotic delivery system for the treatment of osteomyelitis.

An injectable osteoconductive polyelectrolyte complex (PEC)-hydroxyapatite (HAP) formulation capable of controlled delivery of ciprofloxacin has been developed from a novel biodegradable PEC and antibiotic loaded nascent hydroxyapatite (n-HAP) for the treatment of osteomyelitis. A single source (chitosan) derived polyelectrolytes were complexedin situin the presence of n-HAP, pre-loaded with ciprofloxacin. The PEC-(n-HAP) nanoformulation (HPEC) was characterized by FT-IR, XRD, TGA and TEM analyses. HPEC combines functionalities of n-HAP (crystallinity and osteoconductivity) as well as PEC (biodegradable hydrophilic electrostatically bound macromolecular network) imparting better control over swelling and degradation kinetics favourable for drug release and transport of micronutrients. MTT assay and cytoskeleton staining (MG-63 cells) established cytocompatibility of HPEC. Early biomimetic mineralization of apatite was manifested under simulated physiological condition with a Ca/P of 1.23 (day 3) and 1.55 (day 6) complimented byin vitrobiomineralization of MG-63 and human osteosarcoma (HOS) cells in a week (Alizarin Red S staining), which was further validated by calcium quantification. Antibacterial efficacy of HPEC has been evaluated by delivery kinetics of ciprofloxacin and by disc diffusion method againstS. aureusandE. coli. The injectable system therefore possesses unique combination of functionalities: osteoconduction enriched with early biomineralization, antibacterial activity and is biodegradable; hence highly suitable for osteomyelitis treatment.

Intrinsically radiopaque and antimicrobial cellulose based surgical sutures from mechanically powerful Agave sisalana plant leaf fibers.

The judicious configuration of a flexible radiopaque suture would be exemplary to facilitate effortless tracking and precise diagnosis of the sutured surgical site by various X-ray assisted imaging modalities and simultaneously serve as a complementary tool for monitoring the fate of the suture material during the post-operative course. A unique radiopaque cellulose based surgical suture (RF) with good mechanical properties was developed by strategically controlled mercerization and bleaching of mechanically strong natural cellulosic fibers extracted from Agave sisalana plant leaves followed by the facile dip-coating of SrO integrated polylactic acid (PLA). RF exhibited admirable straight-pull tensile strength (184 MPa) and commendable contrast enhancement (277.4%) under digital X-ray radiographic imaging which was further validated by micro-CT analysis. Further, RF has a controlled hydrolytic degradation profile favorable for surgical suturing (mass loss ∼22% in 28 days). The microporous surface architecture of RF (pore size < 10 µm) as a result of SrO-PLA coating enabled the loading of antibiotic (ciprofloxacin) deep inside the pores with a cumulative release of 24% at 28 days under physiological conditions substantiating its feasibility to be used as an efficient antimicrobial suture (CRF) that prevents possible bacterial infections at the surgical site. This has been demonstrated by antibacterial disc diffusion assay performed against two Gram-positive and two Gram-negative bacterial strains. Significantly, both RF and CRF are highly biocompatible as confirmed by MTT assay and F-actin staining. Hence, CRF would be a good biocompatible suture candidate holding good tensile properties, exceptional antimicrobial property and intrinsic radiopacity retention for a period >28 days.

Hierarchically Porous Osteoinductive Poly(hydroxyethyl methacrylate-co-methyl methacrylate) Scaffold with Sustained Doxorubicin Delivery for Consolidated Osteosarcoma Treatment and Bone Defect Repair.

A bifronted cure system for osteosarcoma, a common aggressive bone tumor, is highly in demand to prevail the postsurgical adversities in connection with systemic chemotherapy and repair of critical-size bone defects. The hierarchically porous therapeutic scaffolds presented here are synthesized by free radical-initiated copolymerization of hydroxyethyl methacrylate and methyl methacrylate [HEMA/MMA 80:20 and 90:10 mM, H2O/NaCl porogen], which are further surface-phosphorylated [P-PHM] and transformed to bifunctional by impregnating doxorubicin (DOX) [DOXP-PHM]. The P-PHM scaffolds exhibited porous microarchitecture analogous to native cancellous bone (scanning electron microscopy analysis), while X-ray photoelectron spectroscopy analysis authenticated surface phosphorylation. Based on pore characteristics, swelling attributes and slow-pace degradation, P-PHM9163 and P-PHM8263 (HEMA/MMA 90:10 and 80:20 with H2O/NaCl: 60/3.0 weight %, respectively) were chosen from the series and evaluated for osteoinductive efficacy in vitro. Both P-PHM9163 and P-PHM8263 invoked calcium phosphate mineralization in simulated physiological conditions (day 14) with Ca/P ratios of 1.58 and 1.66 respectively, comparable to human bone (1.67). Early biomineralization (Alizarin Red S and von Kossa staining) was evidenced at day 7, while osteoblast differentiation was verified by time-dependent expression of the typical late marker, osteocalcin, at day 14 and 21 in rat bone marrow mesenchymal cells. DOX-loaded P-PHM9163 (DOXP-PHM9163) exhibited pH-responsive (tumor analogous pH; 6.5) sustained release of DOX for prolonged time (up to 45 days) and invoked cellular alterations by cortical stress fiber formation and DNA fragmentation in human osteosarcoma cells leading to early apoptosis (24 h), validated by annexin V/PI staining (FACS) and immunostaining (F-actin/DAPI). Subsequent to DOX release tenure, the scaffold induced the formation of well-organized, porous post-release Ca-P apatite coating (Ca/P is 1.3) in simulated body fluid (day 14) which further endorses the dual functionality of the system. Altogether, the results accentuate that DOXP-PHM9163 is a potential bifunctional therapeutic scaffold capable of extended localized chemotherapeutic delivery in-line with inherent osteogenesis for efficient bone cancer treatment.

Active bayerite underpinned Ag2O/Ag: an efficient antibacterial nanohybrid combating microbial contamination.

Active surfaces with bactericidal properties are of paramount importance in health care sector as a judicious approach to confront prevalent challenges presented by disastrous pathogenic infections and antibiotic-resistant microbes. Herein, we present Bayerite underpinned Ag2O/Ag (ALD), a nanohybrid with excellent antibacterial and antibiofilm functionalities against tested standard strains and clinical isolates. The multicomponent system coexists and complement each other with respect to phase and functionalities, demonstrated by XRD, XPS, and TEM analyses. In situ reduction of Ag+ ions to Ag0 over Bayerite as a stable bound phase is favoured by pH of the reaction, yielding 60-80% bound Ag protruding outwards facilitating active surface for interaction with microbes. ALD has a minimum inhibitory concentration (MIC) of 0.068 mg/ml against clinical isolates: Pseudomonas aeruginosa RRLP1, RRLP2, Acinetobactor baumannii C78 and C80. Disc diffusion assay demonstrated excellent antibacterial activity against standard strains (positive control: standard antibiotic disc, Amikacin). ALD incorporated PMMA films (5 and 10 wt%; PALD-5 and PALD-10) exhibited significant contact killing (99.9%) of clinical isolates in drop-test besides strong antibacterial activity (disc diffusion assay) comparable to that of ALD. ALD exemplified a dose (0.034 and 0.017 mg/ml) dependent biofilm inhibition (P < 0.001) and significant eradication of pre-formed biofilms (P < 0.001) by clinical isolates. PALD 5 and PALD 10 significantly declined the number of viable biofilm associated bacteria (99.9%) compared to control. Both ALD and PALD samples are proposed as green antibacterial materials with antibiofilm properties. Results also present ample opportunity to explore PALD as antibacterial and/or antibiofilm coating formulations.

Ultrastructural evaluation of in vitro mineralized calcium phosphate phase on surface phosphorylated poly(hydroxy ethyl methacrylate-co-methyl methacrylate).

The in vitro functionality of surface phosphorylated poly(hydroxy ethyl methacrylate-co-methyl methacrylate), poly(HEMA-co-MMA) to induce bioinspired mineralization of calcium phosphate phase is evaluated. The primary nucleation of calcium phosphate on the surface phosphorylated copolymer occurs within 3 days of immersion when immersed in 1.5x simulated body fluid and the degree of mineralization is proportional to the hydroxy ethyl methacrylate content in the copolymer. The calcium phosphate phase is identified as hydroxyapatite by X-Ray diffraction analysis. The transmission electron microscopic evaluation combined with selected area diffraction pattern and energy dispersive analysis exemplified that the primary nuclei of amorphous calcium phosphate transforms to crystalline needle like calcium rich apatite, within a period of 3 days immersion in simulated body fluid. The atomic force microscopic results corroborate the c-axis growth of the crystals within 3 days immersion in SBF.

Surface-transformed osteoinductive polyethylene terephthalate scaffold as a dual system for bone tissue regeneration with localized antibiotic delivery.

Multifunctional scaffolds have recently attained superior significance in tissue regeneration due to combinational activity profile that is usually accomplished by separate sequential therapy. Here, we present a dual system comprised of surface-phosphorylated PET fibrous matrix coated with ciprofloxacin-impregnated biodegradable polymer poly (hydroxyethyl methacrylate) aiming at regeneration of bone tissue deprived of bacterial infections, particularly osteomyelitis. The ATR, XPS and FESEM/EDX results provided confirmative evidences for surface phosphorylation of PET and in situ coating of the polymer. Swelling and contact angle measurements demonstrated improved hydrophilicity which is further corroborated by in vitro degradation profile in PBS. Preliminary evaluation by MTT and actin staining proved its biocompatibility while enhanced in vitro mineralization in 1.5X SBF by FESEM/EDX clearly indicate the primary nucleation and secondary growth of beautiful apatite crystals with Ca/P ratio similar to human bone. Alizarin red S and von Kossa staining validated the biomineralization in MG-63 cells. The sequential expression of early and late biomarkers -alkaline phosphatase (ALP) and osteocalcin (OSN)- of osteoblast differentiation in rat bone marrow mesenchymal cells (BMC) has demonstrated osteoinductive nature of the system. The second functionality of the scaffold has been proven by step-wise ciprofloxacin-release profile (in vitro) with ~60% release within 120 h. In addition, antibacterial studies of ciprofloxacin- eluted from the scaffold have shown apparent zones of inhibition against Staphylococcus aureus (3.6 ± 0.3 cm) and Escherichia coli (3.0 ± 0.8 cm). Hence, the surface-transformed PET scaffold function as a dual system as localized antibiotic delivery vehicle against bone infections and undergo self-biomineralization leading to osteoinduction.

Hydroxyapatite moldable formulation using natural rubber latex as binder.

A simple but efficient processing method for shaping intricate bioceramic green bodies has been developed by using natural rubber latex as binder. Different shapes of hydroxyapatite Ca10(PO4)6(OH)2 (HAP) were molded from a composite formulation containing wet precipitated HAP, natural rubber latex (NRL), and a stabilizer. On controlled heat treatment followed by sintering, dense shapes of HAP contours were obtained. The thermal degradation profile of HAP-NRL composites shows that NRL degrades slowly without any abrupt exotherm. The results of energy dispersive X-ray analysis together with inductively coupled plasma (ICP) analysis indicate that the inorganic residue of NRL does not contain any heavy element. The sintered density of the samples increased with increased HAP content in the formulation and percentage shrinkage reduced accordingly. On varying the HAP content in the formulation from 35 to 95 wt %, the compositions with 85, 90, 92, and 95 wt % HAP showed better flexural strength in the range 40-54 MPa and a flexural modulus value in the range 36-50 GPa. The fracture morphology, as observed by the scanning electron microscope confirms that with increased HAP content in the formulation the sample microstructure attains higher uniformity. The Vickers microhardness for the samples sintered at two different temperatures (1150 and 1250 degrees C) showed that hardness increases with increase in the sintering temperature with a maximum for the highest HAP loaded formulation.

Human osteosarcoma cell adhesion behaviour on hydroxyapatite integrated chitosan-poly(acrylic acid) polyelectrolyte complex.

A novel degradable composite system has been prepared by integrating hydroxyapatite, Ca(10)(PO(4))(6)(OH)(2), (HAP) in a polyelectrolyte complex matrix of chitosan (CHI) and poly(acrylic acid) (PAA). The composite was formulated by integrating 80 wt.% HAP in the polyelectrolyte complex matrix of CHI and PAA in the ratio 40/60 (designated as CPH). The composite could be easily fabricated into clinically significant shapes by a simple moulding procedure intended for bone graft applications. The adhesion behaviour of human osteosarcoma (HOS) cells on this degradable composite system was studied by selecting the polyelectrolyte complex, CHI/PAA 40/60 (designated as CP) as control sample. Light microscopic observations show that cells around CPH retained the typical morphology of HOS cells while cells around the polyelectrolyte complex showed a cytotoxic effect. The adhesion behaviour as well as morphological responses of the seeded cells was further investigated by scanning electron microscopy. The scanning electron micrographs of the polyelectrolyte complex, CP, showed the presence of rounded cells with raised nuclear regions, indicating delayed spreading; cells adhered on CPH were flattened with filopodia and showed good attachment and spreading, indicating better adhesion onto the HAP integrated composite. Comparing the MTT assay for quantitative evaluation of cell viability, CPH showed a higher percentage of metabolically active cells compared to CP.

Aniosotropically organized LDH on PVDF: a geometrically templated electrospun substrate for advanced anion conducting membranes.

A bioinspired geometric templating of an electrospun PVDF substrate with hexagonal platelets of Mg-Al layered double hydroxide (LDH), an intrinsic anion conductor, is presented. The distinctive morphology restructures the internal pore geometry and modulates the dynamic wetting profile of PVDF, transforming it into a highly functional substrate for SAFC anion conducting membranes. The membrane fabricated with PVDF-LDH substrate exhibited exceptionally high durability (>140 °C), high anionic conductivity, ion exchange capacity (IEC), restricted swelling, and improved tensile strength, overcoming critical challenges associated with PVDF electrospun substrates and validating its immense potential as a high-temperature-stable and durable substrate for advanced fuel cell membrane applications.

The secreted protein acidic and rich in cysteine (SPARC) induces endoplasmic reticulum stress leading to autophagy-mediated apoptosis in neuroblastoma.

Our previous studies showed that overexpression of secreted protein acidic and rich in cysteine (SPARC) induced autophagy-mediated apoptosis in PNET cells. In the present study, we attempted to elucidate the molecular mechanisms and signaling cascades associated with SPARC overexpression in combination with radiation therapy that eventually leads to autophagy-mediated apoptosis in neuroblastoma. SPARC expression in SK-N-AS and NB-1691 cells demonstrated the activation of caspase 3, cleavage of PARP and induction of apoptosis. The experiments to unravel the mechanisms associated with autophagy-apoptosis illustrated that SPARC overexpression triggered endoplasmic reticulum (ER) stress and thereby unfolded protein response (UPR). This was apparent with the activation of stress receptors, inositol-requiring enzyme (IRE 1α), RNA-dependent protein kinase (PKR)-like ER kinase (PERK) and BiP. This study further demonstrated the induction of transcription factor CHOP as a result of IRE-JNK activation in response to increased SPARC levels. Inhibition of ER stress and JNK activation led to inhibition of autophagy-mediated apoptosis. Further, the apparent expression of ER stress molecules among the orthotopic tumors treated by SPARC overexpression plasmids substantiated our in vitro observations. Taken together, these results illustrate the critical role of ER stress in regulating autophagy-mediated apoptosis in SPARC-overexpressed neuroblastoma cells and radiation treatment.

SPARC overexpression inhibits cell proliferation in neuroblastoma and is partly mediated by tumor suppressor protein PTEN and AKT.

Secreted protein acidic and rich in cysteine (SPARC) is also known as BM-40 or Osteonectin, a multi-functional protein modulating cell-cell and cell-matrix interactions. In cancer, SPARC is not only linked with a highly aggressive phenotype, but it also acts as a tumor suppressor. In the present study, we sought to characterize the function of SPARC and its role in sensitizing neuroblastoma cells to radio-therapy. SPARC overexpression in neuroblastoma cells inhibited cell proliferation in vitro. Additionally, SPARC overexpression significantly suppressed the activity of AKT and this suppression was accompanied by an increase in the tumor suppressor protein PTEN both in vitro and in vivo. Restoration of neuroblastoma cell radio-sensitivity was achieved by overexpression of SPARC in neuroblastoma cells in vitro and in vivo. To confirm the role of the AKT in proliferation inhibited by SPARC overexpression, we transfected neuroblastoma cells with a plasmid vector carrying myr-AKT. Myr-AKT overexpression reversed SPARC-mediated PTEN and increased proliferation of neuroblastoma cells in vitro. PTEN overexpression in parallel with SPARC siRNA resulted in decreased AKT phosphorylation and proliferation in vitro. Taken together, these results establish SPARC as an effector of AKT-PTEN-mediated inhibition of proliferation in neuroblastoma in vitro and in vivo.

Bioinspired mineralization and cell adhesion on surface functionalized poly(vinyl alcohol) films.

Poly(vinyl alcohol) (PVA) films, when surface functionalized by phosphorylation, induced biomimetic nucleation and growth of calcium phosphate in a simulated physiological environment. The surface phosphorylation on PVA was ensured by attenuated total reflectance infrared spectroscopy. The morphology of the calcium phosphate phase grown on surface-phosphorylated PVA (PPVA) was analysed using scanning electron microscopy coupled with an energy-dispersive X-ray detector. The primary nucleation of calcium phosphate occurs in 3 days and secondary nucleation occurs after 10 days. The energy-dispersive X-ray analysis shows that the Ca/P ratio of the coating increases with time of exposure to the simulated physiological fluid and reaches 1.67 at 10 days. The PPVA supports in vitro cell adhesion and promotes in vitro biomineralization in the presence of cells, evaluated using human osteosarcoma cells.