Conversion of finished leather waste incorporated with plant fibers into value added consumer products - An effort to minimize solid waste in Ethiopia.

Presently, the leftovers from leather product industries are discarded as waste in Ethiopia. The objective of the present study was therefore, to prepare composite sheets by incorporating various plant fibers like enset (Ensete ventricosum), hibiscus (Hibiscus cannabinus), jute (Corchorus trilocularis L.), palm (Phoenix dactylifera) and sisal (Agave sisal) in various proportions into the leather waste. Resin binder (RB) and natural rubber latex (NRL) were used as binding agents for the preparation of the composite sheets. The composite sheets prepared were characterized for their physicochemical properties (tensile strength, elongation at break, stitch tear strength, water absorption, water desorption and flexing strength). Composite sheets prepared using RB having 10% hibiscus, 20% palm and 40% sisal fibers showed better mechanical properties than their respective controls. In composite sheets prepared using NRL having 30% jute fiber exhibited better mechanical properties than its control. Most of the plant fibers used in this study played a role in increasing the performance of the sheets. However, as seen from the results, the contribution of these plant fibers on performance of the composite sheets prepared is dependent on the ratio used and the nature of binder. The SEM studies have exhibited the composite nature of the sheets and FTIR studies have shown the functional groups of collagen protein, cellulose and binders. The prepared sheets were used as raw materials for preparation of items like stiff hand bags, ladies' purse, keychain, chappal upper, wallet, wall cover, mouse pad and other interior decorating products. By preparing such value added products, we can reduce solid waste; minimize environmental pollution and thereby securing environmental sustainability.

Scaffolds containing chitosan, gelatin and graphene oxide for bone tissue regeneration in vitro and in vivo.

Critical-sized bone defects are augmented with cell free and cell loaded constructs to bridge bone defects. Improving the properties of three-dimensional scaffolds with multiple polymers and others is of growing interest in recent decades. Chitosan (CS), a natural biopolymer has limitations for its use in bone regeneration, and its properties can be enhanced with other materials. In the present study, the composite scaffolds containing CS, gelatin (Gn) and graphene oxide (GO) were fabricated through freeze-drying. These scaffolds (GO/CS/Gn) were characterized by the SEM, Raman spectra, FT-IR, EDS, swelling, biodegradation, protein adsorption and biomineralization studies. The inclusion of GO in the CS/Gn scaffolds showed better physico-chemical properties. The GO/CS/Gn scaffolds were cyto-friendly to rat osteoprogenitor cells, and they promoted differentiation of mouse mesenchymal stem cells into osteoblasts. The scaffolds also accelerated bridging of the rat tibial bone defect with increased collagen deposition in vivo. Hence, these results strongly suggested the potential nature of GO/CS/Gn scaffolds for their application in bone tissue regeneration.

A Combinatorial effect of carboxymethyl cellulose based scaffold and microRNA-15b on osteoblast differentiation.

The present study was aimed to synthesize and characterize a bio-composite scaffold containing carboxymethyl cellulose (CMC), zinc doped nano-hydroxyapatite (Zn-nHAp) and ascorbic acid (AC) for bone tissue engineering applications. The fabricated bio-composite scaffold was characterized by SEM, FT-IR and XRD analyses. The ability of scaffold along with a bioactive molecule, microRNA-15b (miR-15b) for osteo-differentiation at cellular and molecular levels was determined using mouse mesenchymal stem cells (mMSCs). miR-15b acts as posttranscriptional gene regulator and regulates osteoblast differentiation. The scaffold and miR-15b were able to promote osteoblast differentiation; when these treatments were combined together on mMSCs, there was an additive effect on promotion of osteoblast differentiation. Thus, it appears that the combination of CMC/Zn-nHAp/AC scaffold with miR-15b would provide more efficient strategy for treating bone related defects and bone regeneration.

Scaffolds containing chitosan/carboxymethyl cellulose/mesoporous wollastonite for bone tissue engineering.

Scaffold based bone tissue engineering utilizes a variety of biopolymers in different combinations aiming to deliver optimal properties required for bone regeneration. In the current study, we fabricated bio-composite scaffolds containing chitosan (CS), carboxymethylcellulose (CMC) with varied concentrations of mesoporous wollastonite (m-WS) particles by the freeze drying method. The CS/CMC/m-WS scaffolds were characterized by the SEM, EDS and FT-IR studies. Addition of m-WS particles had no effect on altering the porosity of the scaffolds. m-WS particles at 0.5% concentration in the CS/CMC scaffolds showed significant improvement in the bio-mineralization and protein adsorption properties. Addition of m-WS particles in the CS/CMC scaffolds significantly reduced their swelling and degradation properties. The CS/CMC/m-WS scaffolds also showed cyto-friendly nature to human osteoblastic cells. The osteogenic potential of CS/CMC/m-WS scaffolds was confirmed by calcium deposition and expression of an osteoblast specific microRNA, pre-mir-15b. Thus, the current investigations support the use of CS/CMC/m-WS scaffolds for bone tissue engineering applications.

Nanohydroxyapatite-reinforced chitosan composite hydrogel for bone tissue repair in vitro and in vivo.

BACKGROUND: Bone loss during trauma, surgeries, and tumor resection often results in critical-sized bone defects that need to be filled with substitutionary materials. Complications associated with conventional grafting techniques have led to the development of bioactive tissue-engineered bone scaffolds. The potential application of hydrogels as three-dimensional (3D) matrices in tissue engineering has gained attention in recent years because of the superior sensitivity, injectability, and minimal invasive properties of hydrogels. Improvements in the bioactivity and mechanical strength of hydrogels can be achieved with the addition of ceramics. Based on the features required for bone regeneration, an injectable thermosensitive hydrogel containing zinc-doped chitosan/nanohydroxyapatite/beta-glycerophosphate (Zn-CS/nHAp/ß-GP) was prepared and characterized, and the effect of nHAp on the hydrogel was examined. METHODS: Hydrogels (Zn-CS/ß-GP, Zn-CS/nHAp/ß-GP) were prepared using the sol-gel method. Characterization was carried out by scanning electron microscopy (SEM), energy dispersive spectroscopy (EDX), Fourier transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD) as well as swelling, protein adsorption, and exogenous biomineralization studies. Expression of osteoblast marker genes was determined by real-time reverse transcriptase polymerase chain reaction (RT-PCR) and western blot analyses. In vivo bone formation was studied using a rat bone defect model system. RESULTS: The hydrogels exhibited sol-gel transition at 37°C. The presence of nHAp in the Zn-CS/nHAp/ß-GP hydrogel enhanced swelling, protein adsorption, and exogenous biomineralization. The hydrogel was found to be non-toxic to mesenchymal stem cells. The addition of nHAp to the hydrogel also enhanced osteoblast differentiation under osteogenic conditions in vitro and accelerated bone formation in vivo as seen from the depositions of apatite and collagen. CONCLUSIONS: The synthesized injectable hydrogel (Zn-CS/nHAp/ß-GP) showed its potential toward bone formation at molecular and cellular levels in vitro and in vivo. The current findings demonstrate the importance of adding nHAp to the hydrogel, thereby accelerating potential clinical application toward bone regeneration.

Antimicrobial behavior of biosynthesized silica-silver nanocomposite for water disinfection: a mechanistic perspective.

The biosynthesis of nano-silica silver nanocomposite (NSAgNC) and it is as antibacterial effect on gram-negative bacteria viz.Escherichia coli and Pseudomonas aeruginosa has been investigated for disinfection of water. The as-synthesized NSAgNC exhibited antibacterial activity in a dose dependent manner and ∼ 99.9% of E. coli and P. aeruginosa were killed at a concentration of 1.5 mg/mL of NSAgNC (5.1 wt% Ag) within 5h. The NSAgNC showed similar antibacterial activities both in oxic and anoxic conditions. The results further demonstrated that NSAgNC exhibited reactive oxygen species (ROS) independent "particle specific" antibacterial activity through multiple steps in absence of leached out Ag(+) ions. The initial binding of NSAgNC on the cell wall caused loss of cell membrane integrity and leakage of cytoplasmic materials. Inhibition of respiratory chain dehydrogenase by NSAgNC caused metabolic inactivation of the cells and affecting the cell viability. Genomic and proteomic studies further demonstrated the fragmentations of both plasmid and genomic DNA and down regulation of protein expression in NSAgNC treated cells, which leading to the cell death. Thus the biosynthesized NSAgNC has great potential as disinfectant for water purification while minimizing the toxic effects.

Evaluation of biomaterial containing regenerated cellulose and chitosan incorporated with silver nanoparticles.

Biomaterials are used in regenerative medicine, implantable materials, controlled release carriers or scaffolds for tissue engineering. In the present study, the composites containing regenerated cellulose (RC) and chitosan (Ch) impregnated with silver nanoparticles (AgNP) with and without antibiotic gentamicin (G) were prepared. The composites prepared were characterized for their physico-chemical and mechanical properties and the results have shown the composite nature. RC-Ch-Ag and RC-Ch-Ag-G composites were used as wound dressing materials in experimental wounds of rats. The healing pattern of the wounds was evaluated by planimetric studies, macroscopic observations, biochemical studies and mechanical properties. The results have shown faster healing pattern in the wounds treated with RC-Ch-Ag and RC-Ch-Ag-G composites compared to untreated control. This study revealed that RC-Ch-Ag composite might be a potential, economical wound dressing material and may be tried on the clinical wounds of animals before being applied on humans.

Qualitative toxicity assessment of silver nanoparticles on the fresh water bacterial isolates and consortium at low level of exposure concentration.

Silver nanoparticles (AgNPs) pose a high risk of exposure to the natural environment owing to their extensive usage in various consumer products. In the present study we attempted to understand the harmful effect of AgNPs at environmentally relevant low concentration levels (≤1ppm) towards two different freshwater bacterial isolates and their consortium. The standard plate count assay suggested that the AgNPs were toxic towards the fresh water bacterial isolates as well as the consortium, though toxicity was significantly reduced for the cells in the consortium. The oxidative stress assessment and membrane permeability studies corroborated with the toxicity data. The detailed electron microscopic studies suggested the cell degrading potential of the AgNPs, and the FT-IR studies confirmed the involvement of the surface groups in the toxic effects. No significant ion leaching from the AgNPs was observed at the applied concentration levels signifying the dominant role of the particle size, and size distribution in bacterial toxicity. The reduced toxicity for the cells in the consortium than the individual isolates has major significance in further studies on the ecotoxicity of the AgNPs.

Poly(ethylene) glycol-capped silver and magnetic nanoparticles: synthesis, characterization, and comparison of bactericidal and cytotoxic effects.

Silver and magnetic (Fe3O4) nanoparticles have attracted wide attention as novel antimicrobial agents due to their unique chemical and physical properties. In order to study the comparative effects on antibacterial and animal cytotoxicity, Staphylococcus aureus and NIH 3T3 fibroblasts were used, respectively. Both nanoparticles were synthesized via a novel matrix-mediated method using poly(ethylene) glycol. Formation of silver nanoparticles was confirmed by fluorescence and ultraviolet-visible spectroscopic techniques. The poly(ethylene) glycol-coated silver and Fe3O4 nanoparticles were characterized by scanning electron microscope, transmission electron microscope, zeta potential, particle size analysis, Fourier-transform infrared, X-ray diffraction, and X-ray photoelectron spectroscopy. The antimicrobial results indicate that both poly(ethylene) glycol-coated silver and Fe3O4 nanoparticles inhibited S. aureus growth at the concentrations of 5 and 10 µg/mL at all time points without showing any significant cytotoxicity on NIH 3T3 fibroblasts. The particle size of both the poly(ethylene) glycol-coated silver and Fe3O4 nanoparticles dominated in the range 10-15 nm, obtained by particle size analyzer. The poly(ethylene) glycol coating on the particles showed less aggregation of nanoparticles, as observed by scanning electron microscope and transmission electron microscope. The overall obtained results indicated that these two nanoparticles were stable and could be used to develop a magnetized antimicrobial scaffolds for biomedical applications.

Collagen based magnetic nanobiocomposite as MRI contrast agent and for targeted delivery in cancer therapy.

BACKGROUND: In this study, an attempt has been made with the advent of technology to prepare a multifunctional nanobiocomposite (NBC) for targeted drug delivery in cancer therapy. METHODS: Collagen (C) was fabricated as nanofibers with multifunctional moieties viz. CFeAb*D by incorporating iron oxide nanoparticles (Fe), coupling with fluorescein isothiocyanate (FITC) labeled antibody (Ab*) and loading an anticancer gemcitabine drug (D). This NBC was characterized by conventional methods and evaluated for its biological activities. RESULTS: The UV-vis and FTIR spectroscopic studies revealed the fluorescein to protein ratio and revealed the presence of iron oxide nanoparticles and their interaction with the collagen molecules, respectively. While SDS-PAGE showed the proteinaceous nature of collagen, VSM and TEM studies revealed magnetic saturation as 54.97emu/g and a magnetic nanoparticle with a diameter in the range of 10-30nm and the dimension of nanofiber ranging from 97 to 270nm. A MRI scan has shown a super paramagnetic effect, which reveals that the prepared NBC can be used as a MRI contrast agent. The MTT assay has shown biocompatibility and an apoptotic effect while phase contrast microscopy exhibited receptor mediated uptake of endocytosis. CONCLUSION: The novelty in the prepared NBC lies in the collagen nanofibers, which have a higher penetrating property without causing much cell damage, biocompatibility and multifunctional properties and is able to carry multifunctional agents. GENERAL SIGNIFICANCE: The study has demonstrated the possible use of CFeAb*D as a multifunctional NBC for biomedical applications.

A novel cross-linked human amniotic membrane for corneal implantations.

The aim of this study was to evaluate the efficacy of Al2(SO4)3 cross-linked human amniotic membrane for ocular surface reconstruction using tissue culture techniques. The human amniotic membrane was cross-linked with Al2(SO4)3, and the cross-linked human amniotic membrane was characterized for its mechanical properties, percentage of swelling in water, sterility, infrared spectroscopy and scanning electron microscopy. The potential of cross-linked human amniotic membrane to support the attachment and proliferation of corneal limbal epithelial cells was assessed in vitro, using static culture system. About 125% increase in the tensile strength was observed in the cross-linked human amniotic membrane compared to human amniotic membrane. Infrared spectroscopy studies have confirmed the cross-linking of human amniotic membrane with Al2(SO4)3. The cross-linked human amniotic membrane was found to be sterile up to 1 year. In culture studies, confluent sheets of epithelial cells were seen at the end of 14th day resembling the morphological features of limbal epithelia. The cross-linked human amniotic membrane has exhibited improved mechanical properties, and the tissue culture studies have shown its feasibility to be used as a limbal transplant. It was concluded that the crosslinked human amniotic membrane with its improved mechanical properties could be used on par with human amniotic membrane.

Biomimetic mineralization of novel silane crosslinked collagen.

Studies on the mechanisms of mineralization of connective tissues, have gained momentum in the recent past. In the present study, Biomimetic mineralization of modified fish scale collagen in vitro is reported. The fish scale collagen was crosslinked with 3-Aminopropyl triethoxysilane and the crosslinked collagen (FCSi) was characterized using conventional methods. The biomimetic mineralization capacity of FCSi was evaluated in SBF solution for 7 days. Formation of hydroxyapatite crystals on the matrix in vitro has been confirmed by FTIR, XRD and SEM-EDAX techniques. The FCSi may be used as a scaffold in bone tissue engineering and as an osteoinductive material in experimental animal models before applying clinically.

Iron nanoparticles from animal blood for cellular imaging and targeted delivery for cancer treatment.

BACKGROUND: Iron nanoparticles (INPs) are usually prepared from inorganic sources, but we have prepared it from goat blood using incineration method. These INPs are then coated with chitosan (C) and coupled with folic acid (F) to form bionanocomposite for folate receptors. METHODS: The bionanocomposite was characterized for its physicochemical properties and cancer cell targeting studies using Fourier transform infrared spectroscopy, transmission electron microscopy, Zeta potential analysis, scanning electron microscopy-energy dispersive X-ray spectroscopy and magnetic resonance imaging analyses. RESULTS: The results have shown that the particle size of the INP-CF was found to be 80-300nm and confirmed the presence of chitosan and folic acid in the bionanocomposite. Cancer and normal mouse embryonic cell line study confirmed the internalization of INP-CF and this phenomenon was also supported by physicochemical studies. CONCLUSION: Thus, nanobiocomposite prepared using natural sources as a raw material will be beneficial compared to commercially available synthetic sources and can be used as receptor targeting agent for cancer treatment. This nanobiocomposite when coupled with substances such as monoclonal antibodies might act as a theranostic nanoagent for cancer therapy in the years to come. GENERAL SIGNIFICANCE: The prepared novel nanobiocomposite containing INPs isolated from natural source may be used as multifunctional agent due its paramagnetic property apart from its drug delivery effect.

Preparation and evaluation of biocomposites as wound dressing material.

Collagen was isolated from the chrome containing leather waste (CCLW) which is a major solid waste in leather industry. Composite films were made using sago starch (SG), soya protein (SY), and collagen (C) and were cross linked with glutaraldehyde (G).The films prepared were characterized for their physico chemical properties like tensile strength, infrared spectra, thermogravimetric analysis, surface morphology, and water absorption studies. Better mechanical properties and surface morphology were observed for SG-SY-G-C films compared to other films prepared using collagen. The composite films prepared were used as wound dressing material on the experimental wounds of rats and healing pattern was evaluated using planimetric, biochemical, and histopathological studies. These studies have revealed better wound healing capacity of SG-SY-G-C film and utilization of CCLW in the preparation of value added product like wound dressing material.

Preparation, characterization and evaluation of biocomposite films containing chitosan and sago starch impregnated with silver nanoparticles.

The positive attributes of excellent biocompatibility and biodegradability of biopolymers with versatile biological activities have provided ample opportunities for further development of functional biomaterials of high potential in various fields. The biopolymers used in this study, i.e. chitosan and sago starch are abundantly available in nature and can be used in various biomedical applications. In the present study, the composite films of chitosan (Ch) and sago starch (SG) impregnated with silver nanoparticles (AgNP) with and without antibiotic gentamicin (G) were prepared by solvent casting method. The films prepared were characterized for their physic-chemical properties using conventional methods. The results obtained showed that with the increase of chitosan content in the composite results in decrease in its water absorption capacity. The FTIR and SEM studies have shown the composite nature of the films prepared. Ch-SG-AgNP and Ch-SG-AgNP-G composites were used as wound dressing materials in experimental wounds of rats. The healing pattern of the wounds was evaluated by planimetric studies, macroscopic observations, biochemical studies and histopathological observations. The results have shown faster healing pattern in the wounds treated with Ch-SG-AgNP and Ch-SG-AgNP-G composites compared to untreated control. This study suggests that Ch-SG-AgNP film may be a potential candidate as a dressing material for wound healing applications.

Evaluation of biocomposite films containing alginate and sago starch impregnated with silver nano particles.

In recent years, the metal nanoparticles/polymer composites have created lot of attraction due to their wide range of applications. In the present study, the composite films of alginate (AL) and sago starch (SG) impregnated with silver nano particles (AgNP) with and without antibiotic gentamicin (G) were prepared by solvent casting method. The films prepared were characterized for thermo gravimetric analysis, SEM, TEM and mechanical properties and the results have shown the composite nature of the films. AL-SG-AgNP and AL-SG-AgNP-G composites were used as wound dressing materials in experimental wounds of rats. The healing pattern of the wounds was evaluated by planimetric studies, macroscopic observations, biochemical studies and histopathological observations. The results have shown faster healing pattern in the wounds treated with AL-SG-AgNP and AL-SG-AgNP-G composites compared to untreated control. This study revealed that AL-SG-AgNP film might be a potential and economical wound dressing material.

Fabrication of novel biofibers by coating silk fibroin with chitosan impregnated with silver nanoparticles.

Nanoparticle based agents often applied as coatings on biomaterials have shown promise in delivering the improved sterility against variety of microbes. In the present study, silk fibers (SF) were coated with chitosan impregnated with silver nanoparticles (Ag-C-SF). These Ag-C-SF fibers were characterized using scanning electron microscopy (SEM) with energy dispersive X-ray analysis, atomic force microscopy (AFM), Infra Red spectroscopy, Thermogravimetric Analysis and Microbiological assay techniques. AFM studies have confirmed the nano sized silver particles in chitosan solution; SEM pictures have exhibited the coating of chitosan along with silver nanoparticles on the silk fibroin. The modified fibers have also shown anti-microbial activity and improved thermal stability. The Ag-C-SF fibers may be explored as wound dressing and tendon reconstruction material in future.

Preparation and partial characterization of collagen sheet from fish (Lates calcarifer) scales.

Fish scales, which are hitherto discarded as waste, were collected and cleaned thoroughly. The scales were hydrolyzed under controlled acidic conditions, neutralized and made in to a sheet, i.e., fish scale collagen sheet (FCS). The FCS was characterized for its infrared spectroscopy (IR), thermo-gravimetric analysis (TGA), scanning electron microscopy (SEM), and mechanical properties. The IR study has shown that the sheet contains both organic and inorganic phases revealing that the scales are partially deminaralized. The tensile strength of FCS is enough if it is used as a wound dressing material. The SEM studies have shown that FCS is porous and exhibited fibrous nature.

Growth of hydroxyapatite on physiologically clotted fibrin capped gold nanoparticles.

The growth of hydroxyapatite (HAp) on physiologically clotted fibrin (PCF)-gold nanoparticles is presented for the first time by employing a wet precipitation method. Fourier transform infrared (FTIR) spectroscopy confirmed the characteristic functionalities of PCF and HAp in the PCF-Au-HAp nanocomposite. Scanning electron microscopy (SEM) images have shown cuboidal nanostructures having a size in the range of 70-300 nm of HAp, whereas 2-50 nm sized particles were visualized in high-resolution transmission electron microscopy (TEM). Energy-dispersive x-ray (EDX) and x-ray diffraction (XRD) studies have confirmed the presence of HAp. These results show that gold nanoparticles with PCF acted as a matrix for the growth of HAp, and that PCF-Au-HAp nanocomposite is expected to have better osteoinductive properties.

Comparative study of some physico-chemical characteristics of osteoporotic and normal human femur heads.

OBJECTIVE: To compare some of the physico-chemical properties of osteoporotic and normal femur heads -- a sample study. MATERIALS AND METHODS: The organic and inorganic parts of human normal (healthy) (N), osteoporotic (OP) femur heads and were separated using conventional methods and their physico-chemical characteristics were compared using infrared spectroscopy (IR), thermogravimetric analysis (TGA), X-ray diffraction (XRD) and scanning electron microscopy (SEM). RESULTS: The data revealed that the extra crosslinking had taken place between the intramolecular alpha chains of collagen of OP bone. This was confirmed by IR spectroscopy and TGA studies. XRD data of the inorganic part of N have shown well-resolved peaks compared to OP revealing the decreased crystallinity in the osteoporotic bone. CONCLUSIONS: The extra intramolecular crosslinking of OP bone collagen molecules increases its fragility. The crystallinity of inorganic phase is less in OP and this may be the reason for its brittleness.