Nickel-ion substituted hydroxyapatite matrices for metal-affinity chromatographic purification of recombinant proteins.

Hydroxyapatite (HAp) is a calcium phosphate ceramic, widely used as a matrix for protein chromatography. The crystal structure of HAp is amenable to a wide range of substitutions, thus allowing for the alteration of its properties. In this study, nickel-ion substituted HAp (NiSHAp) was synthesized using a wet-precipitation method, followed by spray drying. This resulted in the structural incorporation of nickel ions within well-defined microspheres, which were suitable for chromatographic applications. The chromatographic experiments were conducted with NiSHAp and compared with spray-dried hydroxyapatite (SHAp) matrices. Protein purification experiments were conducted using refolded recombinant L-asparaginase (L-Asp), which was produced as inclusion bodies in Escherichia coli. The results showed that NiSHAp effectively adsorbed L-Asp, which was selectively eluted using a phosphate buffer, surpassing the efficiency of imidazole-based elution. In contrast, SHAp showed weaker binding and lower selectivity. The significance of this study lies in developing a scalable NiSHAp matrix for protein purification, especially for large-scale applications. The NiSHAp matrix offers a cost-effective alternative to commercial immobilized metal affinity chromatography (IMAC) adsorbents, especially for purifying His-tagged proteins. This innovative approach exhibits the advantages of mixed-mode chromatography by combining the properties of hydroxyapatite and IMAC in a single matrix, with the potential of improved industrial-scale protein purification.

Enhancing the degradation rate and biomineralization nature of antiferromagnetic Fe-20Mn alloy by groove pressing.

The Fe-Mn alloys are potential candidates for biodegradable implant applications. However, the very low degradation rates of Fe-Mn alloys in the physiological environment are a major disadvantage. In this study, the degradation rate of a Fe-20Mn alloy was improved using the groove pressing (GP) technique. Hot rolled sheets of 2 mm thickness were subjected to GP operation at 1000°C. Uniform fine-grained (UFG) Fe-Mn alloys were obtained using the GP technique. The influence of GP on the microstructure, mechanical properties, degradation behavior in simulated body fluid (SBF), surface wettability, biomineralization, and cytocompatibility was investigated and compared to the annealed (A Fe-Mn) and rolled (R Fe-Mn) sample. The groove-pressed Fe-Mn (G Fe-Mn) alloy had a grain size of approximately 40 ± 16 µm whereas the A Fe-Mn and R Fe-Mn samples had grain sizes of 303 ± 81 and 117 ± 14.5 µm, respectively. Enhanced strength and elongation were also observed with the G Fe-Mn sample. The potentiodynamic polarization test showed the highest Icorr, lowest polarization resistance, and lowest Ecorr for the G Fe-Mn sample among all other samples indicating its higher degradation rate. The weight loss data from immersion tests also shows that the percentage of weight loss increases with time indicating the accelerated degradation behavior of the sample. The static immersion test showed an enhancement in weight loss of 0.46 ± 0.02% and 1.02 ± 0.05% for R Fe-Mn and G Fe-Mn samples, respectively, than A Fe-Mn sample (0.31 ± 0.03%) after 56 days in immersion in SBF. The greater biomineralization tendency in UFG materials is confirmed by the G Fe-Mn sample's stronger hydroxyapatite deposition. When compared to the A Fe-Mn and R Fe-Mn samples, the G Fe-Mn sample has a better wettability, which promotes higher cell adhesion and vitality, showing higher biocompatibility. This study demonstrates that Fe-20Mn processed by GP has potential applications for the manufacture of biodegradable metallic implants.

Injectable macroporous naturally-derived apatite bone cement as a potential trabecular bone substitute.

In this study, we have formulated a novel apatite bone cements derived from natural sources (i.e. eggshell and fishbone) with improved qualities that is, porosity, resorbability, biological activity, and so forth. The naturally-derived apatite bone cement (i.e. FBDEAp) was prepared by mixing hydroxyapatite (synthesized from fishbone) and tricalcium phosphate (synthesized from eggshell) as a solid phase with a liquid phase (a dilute acidic blend of cement binding accelerator and biopolymers like gelatin and chitosan) with polysorbate (as liquid porogen) to get a desired bone cement paste. The prepared cement paste sets within the clinically acceptable setting time (≤20 min), easily injectable (>85%) through hands and exhibits physiological pH stability (7.3-7.4). The pure apatite phased bone cement was confirmed by x-ray diffraction and Fourier transform infrared spectroscopy analyses. The FBDEAp bone cement possesses acceptable compressive strength (i.e. 5-7 MPa) within trabecular bone range and is resorbable up to 28% in simulated body fluid solution within 12 weeks of incubation at physiological conditions. The FBDEAp is macroporous in nature (average pore size ~50-400 µm) with interconnected pores verified by SEM and micro-CT analyses. The FBDEAp showed significantly increased MG63 cell viability (>125% after 72 h), cell adhesion, proliferation, and key osteogenic genes expression levels (up to 5-13 folds) compared to the synthetically derived, synthetic and eggshell derived as well as synthetic and fishbone derived bone cements. Thus, we strongly believe that our prepared FBDEAp bone cement can be used as potential trabecular bone substitute in orthopedics.

Functionally multifaceted alginate/curdlan/agarose-based bilayer fibro-porous dressings for addressing full-thickness diabetic wounds.

Full-thickness diabetic wounds are chronic injuries characterized by bleeding, excessive exude, and prolonged inflammation. Single-layer dressings fail to address their disturbed pathophysiology. Therefore, bilayer dressings with structural and compositional differences in each layer have gained attention. We hypothesized that natural polymer (alginate, curdlan, and agarose) based bilayer dressings with inherent healing properties could effectively resolve these issues. Hence, bilayer dressings were fabricated by electrospinning curdlan/agarose/ polyvinyl alcohol blend (top layer) on an alginate/agarose/polyvinyl alcohol-based lyophilized porous (bottom) layer. Ciprofloxacin was incorporated in both layers as a potential antibacterial drug. The bilayer dressing exhibited high swelling (~1300 %), biocompatibility (>90 % with NIH 3T3 and L929 mouse fibroblasts), and hemocompatibility (hemolysis <5 %). In vitro, scratch assay revealed a faster wound closure (~ 95-100 %) than control. Inhibition zone assay revealed antibacterial activity against Staphylococcus aureus and Escherichia coli. Real-time (in vitro) gene expression experiments performed using human THP-1 macrophages exhibited a significant increase in anti-inflammatory cytokines (4.51 fold in IL-10) and a decrease in pro-inflammatory cytokines (1.42 fold in IL-6) in comparison to lipopolysaccharide. Thus, fabricated dressings with high swelling, hemostatic, immunomodulatory, and antibacterial characteristics can serve as potential multifunctional and sustainable templates for healing full-thickness diabetic wounds.

Fabrication of injectable antibiotic-loaded apatitic bone cements with prolonged drug delivery for treating post-surgery infections.

Antibiotic-loaded bioactive bone substitutes are widely used for treating various orthopedic diseases and prophylactically to avoid post implantation infection. Calcium deficient hydroxyapatite (also known as apatitic bone cement) is a potential bioactive bone substitute in orthopedics due to its chemical composition similar to that of natural bone minerals. In this study, fabrication of mannitol (a solid porogen) incorporated injectable synthetic (Syn) and eggshell derived (ESD) apatitic bone cements loaded with antibiotics (gentamicin/meropenem/ rifampicin/vancomycin) was investigated. The release kinetics of the antibiotics were studied by fitting them with different kinetic models. All the antibiotics-loaded apatitic bone cements set within clinically accepted setting time (20 ± 2 min) and with good injectability (>70%). The antibiotics released from these bone cements were found to be controlled and sustained throughout the study time. Weibull and Gompertz (applies in least initial burst and sustain drug release rate models) were the best models to predict the release behavior. They cements had acceptable compressive strength (6-10 MPa; in the range of trabecular bone) and were biodegradable (21%-27% within 12 weeks of incubation) in vitro in simulated body fluids at physiological conditions. These bone cements showed excellent antibacterial activity from day 1 onwards and no bacterial colony was found from day 3 onwards. The viability of MG63 cells in vitro after 72 h was significantly higher after 24 h (i.e., ~110%). The cells were well attached and spread over the surface of the cements with extended morphology. The ESD antibiotic-loaded apatitic bone cements showed better injectability, degradation and cytocompatibility compared when compared to Syn antibiotic-loaded apatitic bone cements. Thus, we believe that the ESD antibiotic-loaded apatitic bone cements are suitable as potential injectable bone substitutes to avoid post-operative implant associated and other acute or chronic bone infections.

Perspectives of nanofibrous wound dressings based on glucans and galactans - A review.

Wound healing is a complex and dynamic process that needs an appropriate environment to overcome infection and inflammation to progress well. Wounds lead to morbidity, mortality, and a significant economic burden, often due to the non-availability of suitable treatments. Hence, this field has lured the attention of researchers and pharmaceutical industries for decades. As a result, the global wound care market is expected to be 27.8 billion USD by 2026 from 19.3 billion USD in 2021, at a compound annual growth rate (CAGR) of 7.6 %. Wound dressings have emerged as an effective treatment to maintain moisture, protect from pathogens, and impede wound healing. However, synthetic polymer-based dressings fail to comprehensively address optimal and quick regeneration requirements. Natural polymers like glucan and galactan-based carbohydrate dressings have received much attention due to their inherent biocompatibility, biodegradability, inexpensiveness, and natural abundance. Also, nanofibrous mesh supports better proliferation and migration of fibroblasts because of their large surface area and similarity to the extracellular matrix (ECM). Thus, nanostructured dressings derived from glucans and galactans (i.e., chitosan, agar/agarose, pullulan, curdlan, carrageenan, etc.) can overcome the limitations associated with traditional wound dressings. However, they require further development pertaining to the wireless determination of wound bed status and its clinical assessment. The present review intends to provide insight into such carbohydrate-based nanofibrous dressings and their prospects, along with some clinical case studies.

Fabrication and evaluation of agarose-curdlan blend derived multifunctional nanofibrous mats for diabetic wounds.

Diabetic wounds with complex pathophysiology significantly burden the wound care industry and require novel management strategies. In the present study, we hypothesized that agarose-curdlan based nanofibrous dressings could be an effective biomaterial for addressing diabetic wounds due to their inherent healing properties. Hence, agarose/curdlan/polyvinyl alcohol based nanofibrous mats loaded with ciprofloxacin (0, 1, 3, and 5 wt%) were fabricated using an electrospinning technique with water and formic acid. In vitro evaluation revealed the average diameter of the fabricated nanofibers between 115 and 146 nm with high swelling (~450-500 %) properties. They exhibited enhanced mechanical strength (7.46 ± 0.80 MPa -7.79 ± 0.007 MPa) and significant biocompatibility (~90-98 %) with L929 and NIH 3T3 mouse fibroblasts. In vitro scratch assay showed higher proliferation and migration of fibroblasts (~90-100 % wound closure) compared to electrospun PVA and control. Significant antibacterial activity was observed against Escherichia coli and Staphylococcus aureus. In vitro real-time gene expression studies with human THP-1 cell line revealed a significant downregulation of pro-inflammatory cytokines (8.64 fold decrease for TNF-α) and upregulation of anti-inflammatory cytokines (6.83 fold increase for IL-10) compared to lipopolysaccharide. In brief, the results advocate agarose-curdlan mat as a potential multifunctional, bioactive, and eco-friendly dressing for healing diabetic wounds.

Biomimetic ion substituted and Co-substituted hydroxyapatite nanoparticle synthesis using Serratia Marcescens.

Biomimicry is becoming deep-rooted as part of bioceramics owing to its numerous functional advantages. Naturally occurring hydroxyapatite (HA) apart from primary nano structures are also characterised by various ionic substitutions. The ease of accommodating such key elements into the HA lattice is known to enhance bone healing properties of bioceramics. In this work, hydroxyapatite synthesized via biomimetic approach was substituted with individual as well as multiple cations for potential applications in bone repair. Ion substitutions of Sr, Mg and Zn was carried out on HA for the first time by using Serratia grown in a defined biomineralization medium. The individual ions of varying concentration substituted in Serratia HA (SHA) (Sr SHA, Mg SHA and Zn SHA) were analysed for crystallinity, functional groups, morphology and crystal size. All three showed decreased crystallinity, phase purity, large agglomerated aggregates and needle-shaped morphologies. Fourier transform infrared spectroscopy (FTIR) spectra indicated increased carbonate content of 5.8% resembling that of natural bone. Additionally, the reduced O-H intensities clearly portrayed disruption of HA lattice and subsequent ion-substitution. The novelty of this study lies primarily in investigating the co-substitution of a combination of 1% Sr, Zn and Mg in SHA and establishing the associated change in bone parameters. Scanning electron microscope (SEM) and transmission electron microscope (TEM) images clearly illustrated uniform nano-sized agglomerates of average dimensions of 20-50 nm length and 8-15 nm width for Sr SHA; 10-40 nm length and 8-10 nm width for both Zn SHA and Mg SHA and 40-70 nm length and 4-10 nm width in the case of 1% Sr, Zn, Mg SHA. In both individual as well as co-substitutions, significant peak shifts were not observed possibly due to the lower concentrations. However, cell volumes increased in both cases due to presence of Sr2+ validating its dominant integration into the SHA lattice. Rich trace ion deposition was presented by energy dispersive X-ray spectroscopy (EDS) and quantified using inductively coupled plasma optical emission spectrometer (ICP-OES). In vitro cytotoxicity studies in three cell lines viz. NIH/3T3 fibroblast cells, MG-63 osteosarcoma cells and RAW 264.7 macrophages showed more than 90% cell viability proving the biocompatible nature of 1% Sr, Zn and Mg in SHA. Microbial biomineralization by Serratia produced nanocrystals of HA that mimicked "bone-like apatite" as evidenced by pure phase, carbonated groups, reduced crystallinity, nano agglomerates, variations in cell parameters, rich ion deposition and non-toxic nature. Therefore ion-substituted and co-substituted biomineralized nano SHA appears to be a suitable candidate for applications in biomedicine addressing bone injuries and aiding regeneration as a result of its characteristics close to that of the human bone.

Process optimization for the rapid conversion of calcite into hydroxyapatite microspheres for chromatographic applications.

Microsphere hydroxyapatite (HAp) is widely used in various biomedical and chromatographic applications. The work described in this manuscript focuses on a dissolution precipitation method for production of HAp microspheres. This method overcomes certain drawbacks of conventional preparation methods used for HAp preparation, which produce polydisperse particles and are time-consuming and expensive. In the present work, the calcium carbonate (calcite) particles were directly and rapidly converted into HAp microspheres using an inexpensive dissolution precipitation method. The effects of the reaction temperature, time, and mechanical stirring rates were studied, and the reaction parameters were optimized. As confirmed by the XRD studies, the higher reaction temperature and time promote complete HAp conversion, while calcite residues were observed for lower temperatures and times. SEM images show the influence of reaction parameters on the surface microstructure of the microspheres produced. It was observed that the HAp microspheres undergo disintegration at a higher stirring rate. The reaction parameters optimized in this work were ideal for preparing HAp microspheres. The resultant HAp particles were utilized as matrices for chromatographic separation of protein mixtures.

Fabrication and evaluation of multifunctional agarose based electrospun scaffolds for cutaneous wound repairs.

Despite several advances in chronic wound management, natural product based scaffolds with high exude absorption and mechanical strength are still a hotspot in the medical field. Thus, present study illustrates the fabrication of agarose (AG; 10% w/v)/polyvinyl alcohol 12% w/v) based multifunctional nanofibrous electrospun scaffolds. Zinc citrate (1%, 3% and 5% w/w of the polymer) was used as a potential antibacterial agent. The fabricated scaffolds exhibit a swelling of ∼550% in phosphate buffer saline and mechanical strength of 10.11 ± 0.31 MPa which is suitable for most of the wound healing applications that require high strength. In vitro study revealed an increased migration and proliferation of L929 fibroblasts with AG blends when compared to the control. The fabricated scaffolds exhibited antibacterial properties against both Staphylococcus aureus (Gram-positive) and Escherichia coli (Gram-negative) bacterial strains. Hence, a multifunctional (ability to protect wounds from bacterial infections along with effective swelling and mechanical support), natural product based, eco-friendly scaffold to serve as a potential wound dressing material has been successfully fabricated.

A STUDY ON THE RADIATION DOSE DUE TO DISSOLVED 222RN IN WATER SAMPLES OF MANDYA CITY, KARNATAKA, INDIA.

Measurement of dissolved radon concentrations in the water samples collected from selected borewells (depth ~300 ft), wells (depth ~25 ft) and overhead tanks (height ~30 ft) of Mandya city, Karnataka, India, has been carried out by using Emanometry method. The radon concentrations in the waters of borewell, well and overhead tank ranges from 1.5 ± 0.1 to 102.8 ± 5.1, 1.3 ± 0.1 to 3.8 ± 0.4 and 2.5 ± 0.2 to 9.7 ± 1.1 Bq l-1 with the mean values of 16.8, 2.5 and 6.2 Bq l-1, respectively. Majority of borewell water samples showed higher concentrations of dissolved 222Rn compared to waters of well and overhead tank. The overall mean value of dissolved radon concentration of 12.2 Bq l-1 is found to be close to the maximum contaminant level of 11 Bq l-1 suggested by US Environment Protection Agency. The physicochemical parameters like pH, TDS and conductivity were also measured, and dependence of dissolved 222Rn on these parameters has been studied. Using the mean value of dissolved radon concentration, a new attempt has been made to compute the doses imparted to different organs and tissues of the human body. Dosimetric calculations showed that stomach and small intestine receive greater doses due to dissolved radon compared to other organs.

Nanocarrier-based drug delivery systems for bone cancer therapy: a review.

Bone cancer is a malignant tumor that originates in the bone and destroys the healthy bone tissues. Of the various types of bone tumors, osteosarcoma is the most commonly diagnosed primary bone malignancy. The standard treatment for primary malignant bone tumors comprises surgery, chemotherapy and radiotherapy. Owing to the lack of proven treatments, different forms of alternative therapeutic approaches have been examined in recent decades. Among the new therapeutic methodologies, nanotechnology-based anticancer therapy has paved the way for new targeted strategies for bone cancer treatment and bone regeneration. They include approaches such as the co-delivery of multiple drug cargoes, the enhancement of their biodistribution and transport properties, normalizing accumulation and the optimization of drug release profiles to overcome shortcomings of the existing therapy. This review examines the standard treatments for osteosarcoma, their lacunae, and the evolving therapeutic strategies based on nanocarrier-mediated combinational drug delivery systems, and future perspectives for osteosarcoma therapy.

Odontogenic differentiation of inflamed dental pulp stem cells (IDPSCs) on polycaprolactone (PCL) nanofiber blended with hydroxyapatite.

The objective of this study was to develop electrospun polycaprolactone (PCL) membranes blended with hydroxyapatite (HA) and evaluate its potential in differentiating inflamed dental pulp stem/progenitor cells (IDPSCs) into odontoblasts. Electrospun nanofibrous membrane consisting of PCL blended with 10 wt% and 15 wt% of HA were fabricated and the characterization was done by Scanning electron microscopy (SEM), Fourier- transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and contact angle analysis. Cytocompatibility, cell adhesion and odontogenic differentiation ability of the membranes were assessed by MTT, Live/Dead, SEM/DAPI and qPCR studies. The mineral deposition ability of the membranes with IDPSCs was estimated by SEM-EDS. The SEM analysis revealed a nanofibrous texture with an average fiber diameter of 140 nm for PCL, 220 nm for PCL10%HA and 250 nm for PCL15%HA. Among the membranes tested, PCL10%HA favored positive cell attachments, upregulated expression of DSPP and ALP gene and higher Ca/P ratio compared to PCL and PCL15%HA.

Evaluation of effect of two different functionalized nanoparticle photodynamic therapy on nanohardness of root dentin-An in vitro study.

INTRODUCTION: The aim of this study was to evaluate the effect of functionalized nanoparticle photodynamic therapy on Nano hardness of root dentin METHODOLOGY: Fifty single rooted lower premolars were decoronated and sectioned into two halves. Then the samples were embedded horizontally in to the acrylic resin to expose the dentin surface. Baseline nanohardness was done at midroot level using a Nanohardness tester. Exposed dentin surfaces were immersed in the following irrigating solutions Post treatment nanohardness testing was done and results were analyzed statistically RESULTS: In general, all the samples in their respective groups had significant change in nanohardness following immersion in irrigant solutions except in NaOCl + EDTA and saline group. CSRB-np and PLGA-MBnp showed increased nanohardness (P = 0.005 and P = 0.007 respectively). Whereas NaOCl + EDTA + CHX showed decrease in nanohardness (P = 0.04). With regards to Modulus of elasticity (MOE), CSRB-np showed significant difference (P = 0.002) compared to the other groups. MOE increased in CSRB-np and PLGA-MBnp while it decreased in all the other groups. CONCLUSION: In this study, the improvement of nanohardness and modulus of elasticity following the immersion of root dentin in CSRB-np solution was demonstrated.

Combinational delivery of anticancer drugs for osteosarcoma treatment using electrosprayed core shell nanocarriers.

In bone cancer treatment, local delivery of chemotherapeutic agents is preferred compared to other routes of administration. Delivery of multiple drugs using biodegradable carriers improves the treatment efficiency and overcomes drug resistance and toxicity. With this approach, we have developed multilayer biodegradable core shell nanoparticles (NPs) using the electro-spraying technique to deliver methotrexate (MTX) and doxorubicin (DOX) for the treatment of osteosarcoma. These core-shell NPs with a mean particle size of 212 ± 41 nm consist of hydroxyapatite (HA) and DOX as core with the outer shell made of chitosan (CH) followed by polycaprolactone (PCL) with MTX. The encapsulation efficiency of MTX was around 85% and DOX was 38%. In vitro drug release studies were performed in phosphate buffered saline (PBS) at pH 5 and pH 7.4 for 8 days. Different release profiles were observed in both acidic and alkaline pH. The sequential release of MTX followed by DOX was observed in both pH in sustained manner. Human osteosarcoma MG 63 (OMG-63) cells lines were used to test the cytotoxicity of drug loaded NPs. Multi-drug encapsulated bioresorbable and biodegradable electro-sprayed core shell NPs will be promising as a bone substitute for the treatment of osteosarcoma.

Dimethylaminoethyl modified curdlan nanoparticles for targeted siRNA delivery to macrophages.

Curdlan, an insoluble and neutral polysaccharide, was produced from Agrobacterium sp. ATCC 31750 and chemically modified with dimethylaminoethyl (DMAE) group to introduce gene binding ability. The resulting DMAE-curdlan was crosslinked with curdlan nanoparticles using epichlorohydrin. The prepared nanoparticles are spherical with an average diameter of 523 ±â€¯195 nm, stable and are highly biocompatible with differentiated THP-1 macrophages with viability of above 90%. They are taken up more efficiently by RAW 264.7 macrophage cells than by L929 fibroblast cells. They increase the expression of M1 macrophage marker genes, TNFα and CXCL10, and decrease the expression of M2 marker, CD206, indicating their ability to activate M1 phenotype and aid in tumor regression. They are also capable of delivering siRNA to human macrophage-like cells efficiently and inhibit ~59% of the expression of target MMP-9 protein. These results indicate that this modified curdlan-based nanoparticle is a promising vehicle for the delivery of siRNAs to macrophages, which could open up treatment strategies for a range of diseases.

Lower oesophageal peptic stricture after laparoscopic sleeve gastrectomy: World's first case report.

Laparoscopic sleeve gastrectomy (LSG) is the most commonly performed bariatric surgery worldwide. De novo gastroesophageal reflux disease after LSG has been reported in the range of 0%-34.9%. Benign lower oesophageal peptic stricture is rare and has not been reported till date. We present the first case report of benign oesophageal peptic stricture post-sleeve gastrectomy and its management. The management modalities for peptic stricture post-LSG include proton pump inhibitors, endoscopic dilatation and surgical management. Revisional Roux-en-Y gastric bypass along with optimal usage of serial dilatation and medical treatment has been shown to be an effective treatment for the same.

Fatty acids-carotenoid complex: An effective anti-TB agent from the chlorella growth factor-extracted spent biomass of Chlorella vulgaris.

ETHNOPHARMACOLOGICAL RELEVANCE: The multidrug-resistant (MDR) pathogen, Mycobacterium tuberculosis, still remains as one of the major threat to mankind, despite the availability of a live attenuated vaccine and effective antibiotics. Marine microalgae, at all times, act as a key resource for valuable therapeutic compounds with limited side effects. AIM OF THE STUDY: The present explorative attempt is to isolate the biomolecules of pharmacological importance from the marine microalgae, Chlorella vulgaris, and to evaluate its effect on the ever dreadful disease, Tuberculosis. The study is also aimed to develop an economically feasible methodology for by-products extraction from microalgae. MATERIALS AND METHODS: Fatty acids-carotenoid complexes (FACC), namely, FACC-1 (red oil) and FACC-2 (brown oil) were isolated, in addition to lipid and lutein from the Chlorella Growth Factor (CGF, a protein fraction enriched with vitamins, minerals and carbohydrates)-extracted spent biomass through column chromatography. RESULTS: FACC-1 is a complex of fatty acids such as oleic and linoleic acids, and carotenoids such as canthaxanthin and neoxanthin. FACC-2 is a complex of oleic, linoleic and linolenic acids and carotenoids (cryptoxanthin and echinenone). Initial screening for evaluation of minimum bactericidal concentration (MBC) of FACC-1 and -2 was performed against Mycobacterium tuberculosis strains such as H37Rv, SHRE sensitive clinical isolate and SHRE resistant clinical isolate. MBC was noted at 10 µg/mL by FACC-1 and at 5 µg/mL by FACC-2, determined using colony forming and Lucipherase Reporter Mycobacteriophages (LRP) assay. Testing in the PAN sensitive isolates indicated that the MBC was noted at 5 µg/mL by FACC-1 and at 2.5 µg/mL by FACC-2. Complete inhibition (100%) was observed at 100 µg/mL by FACC-1 and at 50 µg/mL by FACC-2. Testing of FACC-1 and FACC-2 individually as well as in combination on two different types of MDR strains confirmed the efficacy of the algal oils, wherein in MDR-strain 1, FACC-1 revealed 50% inhibition at 10 µg/mL, while FACC-2 exhibited the same at 5 µg/mL. Conversely, in the case of MDR strain-2, MBC of FACC-1 was at 500 µg/mL and MBCof FACC-2 to be at 250 µg/mL. No significant synergistic effect was observed on combining both the oils. CONCLUSION: The study signifies the development of a potent therapeutic agent comprising of a complex of anti-TB agent (fatty acids) and antioxidants (carotenoids) from the CGF-extracted spent biomass of C. vulgaris.

Eggshell derived brushite bone cement with minimal inflammatory response and higher osteoconductive potential.

Brushite cements are known for excellent osteoconductive and degradation properties, however, its widespread use is limited due to rapid setting time and poor mechanical properties. The eggshell derived calcium phosphates exhibits improved physical and biological properties due to the presence of biologically relevant ions. In this study, eggshell derived brushite cement (EB) was fabricated using ß-tricalcium phosphate synthesized from eggshells. The presence of trace elements in EB prolonged its setting time. The size of brushite crystals in EB was found to be smaller than the pure brushite cement (PB) leading to increased initial compressive strength and higher in vitro degradation rate. The L6 and MG63 cell lines exhibited good biocompatibility with the cement at the end 72 h. In vivo studies of the cements were performed in rat calvarial defect model. Micro CT analysis showed faster degradation and accelerated bone formation in EB filled defect. Histological studies revealed infiltration of inflammatory cells into the implant site for both the cements till 6th week. However, inflammation was found to be significantly reduced at the 12th week in EB compared to PB leading to complete bone bridge formation. Multi-ion substituted EB seems to be a potential bone substitute material with a reasonable setting time for ease of handling, higher mechanical strength, minimal inflammatory response and higher bone regeneration.

Theranostic Calcium Phosphate Nanoparticles With Potential for Multimodal Imaging and Drug Delivery.

Calcium phosphate (CaP) bioceramics closely resemble the natural human bone, which is a main reason for their popularity as bone substitutes. However, this compositional similarity makes it difficult to distinguish CaPs, especially in particulate form, from native bone by imaging modalities such as X-ray radiography, computed tomography (CT), and magnetic resonance imaging (MRI) to monitor the healing progress. External contrast agents can improve the imaging contrast of CaPs but can affect their physicochemical properties and can produce artifacts. In this work, we have attempted to improve the contrast of CaP nanoparticles via ion substitutions for multimodal imaging. Calcium-deficient hydroxyapatite (CDHA) nanoparticles with silver (Ag), gadolinium (Gd), and iron (Fe) substitution were prepared by a microwave-accelerated wet chemical process to improve the contrast in CT, T1 (spin-lattice), and T2 (spin-spin) MRI relaxation modes, respectively. Ag, Gd, and Fe were substituted at 0.25, 0.5, and 0.25 at.%, respectively. The ion-substituted CDHA (ICDHA) was found to be phase pure by X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FT-IR). Transmission electron microscopy (TEM) images showed that the ICDHA nanoparticles were platelet shaped and of 52 ± 2 nm length and 6 ± 1 nm width. The ICDHA showed high contrast in X-ray and CT compared to CDHA. The vibrating sample magnetometry (VSM) studies showed the ICDHA to exhibit paramagnetic behavior compared to diamagnetic CDHA, which was further confirmed by improved contrast in T1 and T2 MRI mode. In addition, the in vitro tetracycline drug loading and release was studied to investigate the capability of these nanoparticles for antibiotic drug delivery. It was found that a burst release profile was observed for 24 h with 47-52% tetracycline drug release. The ICDHA nanoparticles also showed in vitro antibacterial activity against Staphylococcus aureus and Escherichia coli due to Ag, which was further enhanced by antibiotic loading. In vitro biocompatibility studies showed that the triple-ion-substituted ICDHA nanoparticles were cytocompatible. Thus, the ion-substituted CDHA nanoparticles can have potential theranostic applications due to their multimodal image contrast, antibacterial activity, and drug delivery potential. Future work will be conducted with actual bone samples in vitro or in animal models.