Multi-functional bandage - bioactive glass/metal oxides/alginate composites based regenerative membrane facilitating re-epithelialization in diabetic wounds with sustained drug delivery and anti-bactericidal efficacy.

Chronic wounds, especially diabetic, foot and pressure ulcers are a major health problem affecting >10 % of the world's populace. Calcium phosphate materials, particularly, bioactive glasses (BG), used as a potential material for hard and soft tissue repair. This study combines nanostructured 45S5 BG with titania (TiO2) and alumina (Al2O3) into a composite via simple sol-gel method. Prepared composites with alginate (Alg) formed a bioactive nanocomposite hydrogel membrane via freezing method. X-ray diffraction revealed formation of two phases such as Na1.8Ca1.1Si6O14 and ß-Na2Ca4(PO4)2SiO4 in the silica network. Fourier transformed InfraRed spectroscopy confirmed the network formation and cross-linking between composite and alginate. <2 % hemolysis, optimal in vitro degradation and porosity was systematically evaluated up to 7 days, resulting in increasing membrane bioactivity. Significant cytocompatibility, cell migration and proliferation and a 3-4-fold increase in Collagen (Col) and Vascular Endothelial Growth Factor (VEGF) expression were obtained. Sustained delivery of 80 % Dox in 24 h and effective growth reduction of S. aureus and destruction of biofilm development against E. coli and S. aureus within 24 h. Anatomical fin regeneration, rapid re-epithelialization and wound closure were achieved within 14 days in both zebrafish and in streptozotocin (STZ) induced rat in vivo animal models with optimal blood glucose levels. Hence, the fabricated bioactive membrane can act as effective wound dressing material, for diabetic chronic infectious wounds.

Chitosan bioactive glass scaffolds for in vivo subcutaneous implantation, toxicity assessment, and diabetic wound healing upon animal model.

This study aims to develop chitosan-bioactive glass (BG) scaffolds for diabetic wound healing, toxicity valuation, and subcutaneous implantation in animals for biocompatibility assessment. The scaffolds were prepared by lyophilization technique. In specific BG without sodium (Na), composited with chitosan for better biological activities. The equipped scaffolds were studied for their physiochemical, biological, in vitro and in vivo performances. The chitosan and chitosan-BG (Na free) scaffolds show reliable biocompatibility, cytocompatibility, anti-oxidant, and tissue regeneration. The biocompatibility, toxicity assessments, and diabetic skin wound healing experiments were examined through in vivo studies using Sprague Dawley rats. The extracted tissue samples were analyzed using hematoxylin-eosin- (H and E) and Masson's trichrome staining. Further, tissue excised after scaffold implantation declared non-toxic, non-allergic, and anti-inflammatory nature of chitosan scaffolds. Moreover, the total ribonucleic acid (RNA) expression levels were measured using reverse transcription-polymerase chain reaction (RT-PCR) for the scaffolds against vascular endothelial growth factor (VEGF), and collagen type one (Col-1) primers. Admirably, the scaffolds achieved the best level of skin wound healing via tissue regeneration by increasing epithetical cell formation and collagen deposition. Thus, the biocompatibility, non-toxicity, anti-inflammatory, and wound healing efficiency proved that the chitosan-BG (Na free) scaffold can be readily substantial for wound healing.

Unveiling pro-angiogenesis and drug delivery using dual-bio polymer with bio-ceramic based nanocomposite hydrogels.

In regenerative medicine, blood vessel development is of utmost importance as it enables the restoration of blood flow to tissues, and facilitate rapid vascularization in clinical tissue-engineered grafts. Herein, we fabricate the nanocomposite hydrogels from BG (clinophosinaite), alginate, Polyethylene glycol (PEG) and Dexamethasone (DEX) for the dual applications of drug delivery and angiogenesis assay. The hydrogels were fabricated through cross-linking approach and termed as alginate/PEG (A), alginate/PEG/clinophosinaite (AC), and alginate/PEG/clinophosinaite/DEX (ACD) that further subjected to structural characterization, using powder X-ray diffraction, and fourier-transform infrared spectroscopy. Porous nanostructures and sheets were imaged using field emission scanning electron microscopy (FESEM), which aid in nutrient and oxygen transport to support angiogenesis. The nanocomposite hydrogels evidently demonstrated good hemocompatibility and fully hydrophilic (30.20°). By means of liquid displacement technique, the nanocomposite hydrogel achieves 47% of porosity with the compressive strength about 0.04 MPa. In alginate/PEG/clinophosinaite and alginate/PEG/clinophosinaite/DEX systems, water absorption capacity reached 85% in 6 h and maintained 90% retention after 12 h. Further, leachable tests revealed that the hydrogel had not deformed even after 24 h. In vitro drug release studies evidently divulge sustainable delivery of DEX from alginate/PEG/clinophosinaite/DEX hydrogel with superior characteristics for drug release. The angiogenesis assay also evidently revealed that the AC and ACD hydrogels, demonstrated higher angiogenic properties with, promoted blood vessel development.

3D interconnected porous PMMA scaffold integrating with advanced nanostructured CaP-based biomaterials for rapid bone repair and regeneration.

Bioactive scaffolds with polymer and nanostructured bioactive glass-based composites are promising materials for regenerative applications in consequence of close mimics of natural bone composition. Poly methyl methacrylate (PMMA) is a highly preferred thermoplastic polymer for orthopedic applications as it has good biocompatibility. Different kinds of bioactive, biodegradable as well as biocompatible biomaterial composites such as Bioglass (BG), Hydroxyapatite (Hap), and Tricalcium phosphate (TCP) can be integrated with PMMA, so as to augment the bioactivity, porosity as well as regeneration of hard tissues in human body. Among the bioactive glass, 60S BG (Bioactive glass with 60 percentage of Silica without Sodium ions) is better materials among aforementioned systems owning to mechanical stability as well as controlled bioactive material. In this work, the fabrication of PMMA-CaP (calcium phosphate)-based scaffolds were carried out by Thermal Induced Phase Separation method (TIPS). X-ray diffractogram analysis (XRD) is used to examine the physiochemical properties of the scaffolds that evidently reveal the presence of calcium phosphate besides calcium phosphate silicate phases. The Field Emission Scanning Electron Microscopy (FESEM) studies obviously exhibited the microstructure of the scaffolds as well as their interconnected porous morphology. The PMMA/60S BG/TCP (C50) scaffold has the maximum pore size, measuring 77 ± 23 µm, while the average pore size ranges from 50 ± 20 to 80 ± 23 µm. By performing a liquid displacement method, the C50 scaffold is found to have the largest porosity of 50%, high hydrophilicity of 118.16°, and a compression test reveals the scaffolds to have a maximum compressive strength of 0.16 MPa. The emergence of bone-like apatite on the scaffold surface after 1st and 21st days of SBF immersion is further supported by in vitro bioactivity studies. Cytocompatibility and hemocompatibility analyses undoubtedly confirmed the biocompatibility behavior of PMMA-based bioactive scaffolds. Nano-CT investigation demonstrates that PMMA-CaP scaffolds provide more or less alike morphologies of composites that resemble the natural bone. Therefore, this combination of scaffolds could be considered as potential biomaterials for bone regeneration application. This detailed study promisingly demonstrates the eminence of the unique scaffolds in the direction of regenerative medicines.

Bioceramic and polycationic biopolymer nanocomposite scaffolds for improved wound self-healing and anti-inflammatory properties: an in vitro study.

The development of wound healing scaffolds with high porosity, rapid healing properties, and anti-inflammatory functionality is vital in the chronic wound healing stage for the production of extracellular matrices of injured tissues. The 45S5 bioactive glass (BG) possesses good biocompatibility and provides a potential bonding resource for fibroblast cell proliferation, growth factor synthesis, and granulated tissue formation. Chitosan, a natural polymer, promotes tissue regeneration and has anti-microbial properties. BG and chitosan scaffolds were prepared by the freeze-drying (lyophilization) method. The chitosan scaffold is a semi-crystalline polymer with a random crystal structure because it contains more hydroxyl groups. Chitosan alone shows a sheet-like morphology with a porous microstructure (1.7475 nm). BG particulates were well decorated over the surface of the chitosan scaffold with a homogeneous dispersion. Cell viability was observed for L929 cells on the chitosan-BG scaffolds. Confocal images vividly depict the interaction of the L929 cells with the scaffold without causing any damage to the cell membrane. In vitro scratch assay shows the best wound healing activity (complete wound closure) for the BG-chitosan nanocomposite scaffolds at 18 h. The chitosan-BG scaffolds were combined with anti-inflammatory drugs and induced inflammatory genes at an inhibition rate of COX of (36, 28, and 30%), LOX of (20, 13, and 14%), and NO of (48, 38, and 39%) for chitosan, chitosan-BG, and chitosan-BG (Na-free) at 100 µL addition. The in vitro bioactivities proved that the chitosan-BG scaffolds could enable better cell formation, and exhibited improved biocompatibility, and anti-inflammatory and wound healing properties.

Drug infused Al2O3-bioactive glass coatings toward the cure of orthopedic infection.

A unique implant coated substrate with dual-drug-eluting system exhibiting antibacterial, anti-inflammatory, and bone regenerative capacity has been fabricated using spray pyrolysis deposition (SPD) method. Bioglass (BG) and BG-alumina (BG-Al) composites coatings with different concentrations of Al incorporated on BG network over the Cp-Ti substrate were fabricated using SPD technique. Phase purity of BG and BG-Al composites were analyzed by XRD in which Na2Ca2Si3O9 and ß-Na2Ca4(PO4)2SiO4) and Na7.15(Al7.2Si8.8O32) phases were formed. Surface morphology of the coated substrates was analyzed by SEM. Uniformity of the coatings were evaluated by surface profilometer and the uniform distribution the nanoparticles were confirmed with Elemental mapping. Systematically, each apatite layer formation on coated substrate was confirmed by immersing the samples for 1, 3, and 7 days in simulated body fluid and the needle-like structure was characterized using SEM. Cumulative release of Tetracycline hydrochloride (Tet) antibiotic and Dexamethasone (Dex) anti-inflammatory drug-loaded BG-Al and BG-Al composite-coated substrate were studied for 24 h. Antibacterial activity of the coated substrates were evaluated by time-dependent growth inhibition and minimal inhibitory concentration (MIC) assays in which BG-Al and BG-Al composite loaded with Tet showed considerable growth inhibition against S. aureus. Osteoblast-like cells (MG-63) exhibited profound proliferation with no cytotoxic effects which was due to release of Dex drug-coated substrates. Thus, surface modification of Cp-Ti substrate with BG, BG-Al composites coatings loaded with Tet and Dex drug can be considered for post-operative orthopedic implant infection application.

Bioactive, degradable and multi-functional three-dimensional membranous scaffolds of bioglass and alginate composites for tissue regenerative applications.

With a worldwide increase in the aged populace and associated geriatric diseases, there is an enormous need for the regeneration of degenerated organ systems. For this purpose, bioactive glass particulate (nBG) integrated alginate (Alg) composite membrane scaffolds were fabricated by a sol-gel assisted freeze-drying method and validated for their multifunctional utility in regenerative medicine. The presence of the combeite highly crystalline structure of nBG and Alg amorphous broad peaks were confirmed. Repetitive peaks from acids along with stretching confirmed the chemical interactions of the composites. Swelling ability, porosity, and in vitro degradation and biomineralization were analysed for up to 7 days. The results indicated that reduced swelling and degradation enhanced apatite formation. Hemocompatibility and the hemostatic properties on scaffolds were also systematically investigated. Additionally, significant cyto-compatibility and proliferation were noted in a culture with KB3-1. Further 3-D co-cultures with HDF cells and KB3-1 cells exhibited spheroid formation on Alg, nBG/Alg and nBG-Zr/Alg with profound dynamism required to establish organoids of interest. Thus, the results indicate that these 3D hydrogel membranes could offer infinite possibilities in the field of regenerative medicine, notably as an extracellular matrix (ECM) supporting the regeneration of bone, intra-vascularization, and neo-tissue formation, such as cartilage and ligaments.

Impact of copper on in-vitro biomineralization, drug release efficacy and antimicrobial properties of bioactive glasses.

This study highlights the incorporation of copper in the bioactive glasses (BAG) network that greatly influences the morphological, structural and biological properties. By increasing the copper incorporation in BAG, increment in cell volume was obtained from XRD patterns, and concomitantly, dominant phosphate bands and latent silica bands were observed by FT-IR and Raman spectroscopic results. The Cu addition also affected particle appearance to vary from spherical to cluster-like cubes in 1.5% and 2.5% copper-doped BAG. Due to the mesoporous network 1.5% and 2.5% copper-doped BAG showed enhanced release of anti-inflammatory drugs such as Acetaminophen (ACE) and Ibuprofen (IBU) in which, the drug release profiles showed best fit with kinetic models of First order, Korsmeyar-Peppas and Higuchi. Copper doping influences the lattice of BAG, as a result morphology and porosity varied, which regulates the ionic dissolution, hence, prompting bioactivity was perceived from 1.5% and 2.5% copper-doped bioactive glasses (Cu-BGs). Moreover, 2.5% Cu-BG and 1.5% Cu-BG showed highest rate of ROS detection, as well as improved antimicrobial activity. This study established that up to certain proportion of copper incorporation in BAG network, potentially enhances the biomineralization and turns the morphology towards minimal size with mesoporous nature. Due to the abundance in oral microbial exposure, copper amplifies the superior antimicrobial properties, and Cu-BGs act as a drug carrier to load ACE and IBU, which potentially up-regulate the healing properties in dental application.

Effect of microwave and probe sonication processes on sol-gel-derived bioactive glass and its structural and biocompatible investigations.

This study is to investigate the effect of synthesis approaches on morphology, porosity, and biocompatibility of bioactive glass (BG). BG prepared through sol-gel approach was subsequently subjected to microwave and probe sonication techniques to investigate the structural and morphological effect. Hexagonal rod-shaped morphology was obtained in sol-gel-derived bioactive glass, whereas mesoporous particles and spherical-shaped morphology were observed in probe-sonicated and microwave-assisted sol-gel approaches, respectively. The probe-sonicated BG has mesopores with pore diameter of 14.7 nm, whereas surface porosity of 1.5 nm, and 3.5 nm for pure sol-gel and microwave-assisted sol-gel fabricated BGs. Granular size, shape, and porosity have a significant role at the point of contact with cellular membrane. Therefore, we studied the biocompatibility with respect to morphology and porosity of the fabricated BGs. From this study, we observed that the BG prepared using probe sonication method controls the particle size, further it enhances the porosity that altogether improves the biocompatibility. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 108B:143-155, 2020.

Effect of Titania Concentration in Bioglass/TiO2 Nanostructures and Its In Vitro Biological Property Assessment.

Bioglass 45S5 (45% SiO2-24.5% NaO-24.5% CaO-6% P2O5) is a unique bioactive material, which is being used for bone and dental substitution. This system has been highly preferred for its osteoconductive and osteoinductive performance. Despite its attractive bioactivity, there are limitations in using this material for orthopedic and dental applications due to its poor processability and mechanical strength. To improve the load-sharing and stress distribution, TiO2 nanoparticles have been introduced into the nanoBioglass (nBG) by sol-gel method. The structural analyses of the samples were confirmed using X-ray diffraction, Raman-spectroscopy and FTIR. The morphologies of the samples were characterized by FESEM. The apatite formation of the nBG/TiO2 composites was investigated by immersing the samples in simulated body Fluid (SBF) solution for 1 and 3 days, which reveals the acceptable compatibility for different concentrations of all the composition. Hemolysis studies of the nanobiomaterials were carried out to understand the interactions of biomaterials with blood which shows 0.2%-2% of lysis which is acceptable as per ASTM standard. Cell culture and cell proliferation studies of bioglass, nBG/TiO2 nanocomposite on MG-63 pre-osteoblast cell line for 24 h, 48 h and 72 h showed 80% to 95% of cell viability. Also, it was found that the nBG/TiO2 bio-nanocomposites containing low content of titania had good bioactivity properties that is comparable to cortical bone. Hence, nBG/TiO2 bio-nanocomposites are greatly promising for medical applications such as bone substitutes especially in load-bearing sites.