Design and characterization of highly porous curcumin loaded freeze-dried wafers for wound healing.

The goal of this research was to evaluate the beneficial effects of topical curcumin loaded freeze-dried wafers in wound healing. Curcumin wafers were fabricated by cross-linking of chitosan with beta glycerophosphate under magnetic stirring. Composite wafers were prepared by the addition of sodium hyaluronate. Wafers were fabricated by freeze-drying technique. The resulted wafers were examined by naked eye and their dimensions were measured using a caliper. % Drug content, in-vitro release and % water uptake tests were conducted to characterize the fabricated wafers. Porosity testing, compressive mechanical behavior, morphological examination using scanning electron microscopy, thermal behavior using differential scanning calorimetry and Fourier transform infrared spectroscopy were all carried out on the optimized cross-linked wafers followed by their microbiological assays and cytotoxicity studies. The results showed that the optimized wafers possessed high water uptake capabilities while entertaining very high porosity levels (86-89%). Microbiological assay revealed the superiority of the selected curcumin wafers versus free curcumin in bacterial growth inhibition against Staphylococcus epidermidis and Staphylococcus aureus (MRSA) bacteria. The anti-inflammatory effects of the selected curcumin wafers were evaluated against pro-inflammatory cytokines. The results suggested that they were significantly better than free curcumin in lowering cytokines levels. To conclude, the obtained findings revealed that curcumin wafers offered a promising solution in the field of wound healing.

Design and Characterization of Spray-Dried Proliposomes for the Pulmonary Delivery of Curcumin.

PURPOSE: The goal was to directly deliver curcumin, a natural polyphenolic anticancer and anti-inflammatory compound, to the lung tissues with minimal systemic exposure through the fabrication of proliposomes, overcoming its poor aqueous solubility and oral bioavailability. METHODS: Nano-spray drying was employed to prepare proliposomes using hydroxypropyl beta-cyclodextrin as a carrier. Lecithin and cholesterol were used as lipids, stearylamine and Poloxamer 188 were added as positive charge inducer and a surfactant, respectively. Different characterization parameters were evaluated like percentage yield, entrapment efficiency, drug loading, aerodynamic particle size, in vitro release besides morphological examination. Cytotoxicity studies on cell line A549 lung tumor cells as well as in vivo lung pharmacokinetic studies were also carried. RESULTS: The optimized formulations showed superior aerosolization properties coupled their enhanced ability to reach deep lung tissues with a high % of fine particle fraction. Cytotoxicity studies using MTT assay demonstrated enhanced growth inhibitory effect on lung tumor cells A549 and significant reduction of proinflammatory cytokines such as tumor necrosis factor-α, interleukin-6 and interleukin-10 compared to the pure drug. Results of lung pharmacokinetic tests confirmed the superiority of proliposomal curcumin over curcumin powder in both, the rate and extent of lung tissue absorption, as well as the mean residence time within the lung tissues. CONCLUSION: The pulmonary delivery of curcumin-loaded proliposomes as dry powder provides a direct approach to lung tissues targeting while avoiding the limitations of the oral route and offering a non-invasive alternative to the parenteral one.

Nanofibrillated cellulose/cyclodextrin based 3D scaffolds loaded with raloxifene hydrochloride for bone regeneration.

This study intended to design novel nanofibrillated cellulose/cyclodextrin-based 3D scaffolds loaded with raloxifene hydrochloride for bone regeneration. The scaffolds were prepared using two different types of cyclodextrins namely; beta-cyclodextrin and methyl-beta-cyclodextrin. The prepared scaffolds were evaluated by characterizing their porosity, compressive strength, in-vitro drug release, FT-IR and XRD as well as their morphological properties using SEM. Results presented that the prepared scaffolds were highly porous, additionally, the scaffold containing drug/beta-cyclodextrin kneaded complex (SC5) showed the most controlled drug release pattern with the least burst effect and reached almost complete release at 480 h. The in-vitro cytocompatibility and regenerative effect of the chosen scaffold (SC5) was assessed using Saos-2 cell line. Results proved that SC5 was biocompatible. Moreover, it enhanced the cell adhesion, alkaline phosphatase enzyme expression and calcium ion deposition which are essential factors for bone mineralization. The obtained observations presented a novel, safe and propitious approach for bone engineering.

Etoricoxib-loaded bio-adhesive hybridized polylactic acid-based nanoparticles as an intra-articular injection for the treatment of osteoarthritis.

Osteoarthritis is a major problem in elder people. Etoricoxib-loaded bio-adhesive hybridized nanoparticles were prepared using polylactic acid (PLA) and chitosan hydrochloride (CS-HCl) in presence of Captex®200 as a liquid oil, polyvinyl alcohol (PVA) and Tween®80 as surfactants. The study aimed to present a new intra-articular treatment of osteoarthritis with anti-inflammatory as well as bone rebuilding effects. Hybridized nanoparticles were fabricated applying the emulsion solvent evaporation technique then assessed for particle size, zeta potential, entrapment efficiency and in-vitro drug release. Furthermore, FT-IR and DSC in addition to morphological examination were done. Results revealed that the formulation composed of PLA:Captex®200 in ratio 1:2 (w/w), 1%w/v Tween®80, 0.3% w/v CS-HCl and 3%w/v PVA possessed the smallest particle size and the most sustained drug release, thus was sorted for further analyses. The selected formulation ability to interact with the negatively charged sodium fluroscein was evaluated to predict its binding with the naturally occurring hyaluronic acid in the knee joint where promising results were obtained. Results showed the cytocompatibility of the formulation when tested using MC3T3-E1 normal bone cell line, enhanced ALP activity and increased calcium ion deposition and binding. Results suggested that the presented formulation can be considered as an innovative approach for osteoarthritis.

Risedronate-Loaded Macroporous Gel Foam Enriched with Nanohydroxyapatite: Preparation, Characterization, and Osteogenic Activity Evaluation Using Saos-2 Cells.

The application of minimally invasive surgical techniques in the field of orthopedic surgery has created a growing need for new injectable synthetic materials that can be used for bone grafting. In this work, novel injectable thermosensitive foam was developed by mixing nHAP powder with a thermosensitive polymer with foaming power (Pluronic F-127) and loaded with a water-soluble bisphosphonate drug (risedronate) to promote osteogenesis. The foam was able to retain the porous structure after injection and set through temperature change of PF-127 solution to form gel inside the body. The effect of different formulation parameters on the gelation time, porosity, foamability, injectability, and in vitro degradation in addition to drug release from the prepared foams were analyzed using a full factorial design. The addition of a co-polymer like methylcellulose or sodium alginate into the foam was also studied. Results showed that the prepared optimized thermosensitive foam was able to gel within 1 min at 37°C, and sustain the release of drug for 72 h. The optimized formulation was further tested for any interactions using DSC and IR, and revealed no interactions between the drug and the used excipients in the prepared foam. Furthermore, the ability of the pre-set foam to support osteoblastic-like Saos-2-cell proliferation and differentiation was assessed, and revealed superior function on promoting cellular proliferation as confirmed by fluorescence microscope compared to the plain drug solution. The activity of the foam treated cells was also assessed by measuring the alkaline phosphatase activity and calcium deposition, and confirmed that the cellular activity was greatly enhanced in foam treated cells compared to those treated with the plain drug solution only. The obtained results show that the prepared risedronate-loaded thermosensitive foam would represent a step forward in the design of new materials for minimally invasive bone regeneration.

Bioactive injectable triple acting thermosensitive hydrogel enriched with nano-hydroxyapatite for bone regeneration: in-vitro characterization, Saos-2 cell line cell viability and osteogenic markers evaluation.

Hydrogels forming in-situ have gained great attention in the area of bone tissue engineering recently, they were also showed to be a good and less invasive alternative to surgically applied ones. The primal focus of this study was to prepare chitosan-glycerol phosphate thermosensitive hydrogel formed in-situ and loaded with risedronate (bone resorption inhibitor) in an easy way with no requirement of complicated processes or large number of equipment. Then we investigated its effectiveness for bone regeneration. In-situ forming hydrogels were prepared using chitosan cross-linked with glycerol phosphate and loaded with risedronate and nano-hydroxyapatite as bone cement. The prepared hydrogels were characterized by analyzing their gelation time at 37 °C, % porosity, swelling index, in-vitro degradation, rheological properties, and in-vitro drug release. Results showed that the in-situ hydrogels prepared using 2.5% (w/v) chitosan cross-linked with 50% (w/v) glycerol phosphate in the ratio (9:1, v/v) reinforced with 20 mg/mL and nano-hydroxyapatite possessed the most sustained drug release profile. This optimized formulation was further evaluated using DSC and FTIR studies, in addition to their morphological properties using scanning electron microscopy. The effect on Saos-2 cell line viability was evaluated also using MTT assay on the optimized hydrogel formulation in addition to their action on cell proliferation using fluorescence microscope. Moreover, calcium deposition on the hydrogel and alkaline phosphatase activity were evaluated. Risedronate-nano-hydroxyapatite loaded hydrogels significantly enhanced the Saos-2 cell proliferation in addition to enhanced alkaline phosphatase activity and calcium deposition. Such results suggest that risedronate-nano-hydroxyapatite loaded hydrogels present great biocompatibility for bone regeneration. Proliferation of cells, as well as deposition of mineral on the hydrogel, was an evidence of the biocompatible nature of the hydrogel. This hydrogel formed in-situ present a good less invasive alternative for bone tissue engineering.

Mesenchymal stem cells associated with chitosan scaffolds loaded with rosuvastatin to improve wound healing.

In this study we explored the role of rosuvastatin calcium in skin regeneration as statins play important role in the field of tissue engineering. Chitosan hydrochloride was crosslinked with different weight ratios of collagen, ß-glycerolphosphate and carboxymethyl cellulose to produce scaffolds by lyophilization technique. Subsequently, the fabricated scaffolds were examined for their morphology, water absorption capacity, water retention, friability and in-vitro drug release as well as in-vivo studies. The results revealed porous 3-D structured scaffolds with maximum water absorption values-ranging between 396 and 2993%. Scaffolds containing carboxymethyl cellulose revealed highest water absorption-values. In-vitro drug release results showed gradual drug release for 60 h with mean dissolution time-values (MDT) between 13 and 21 h. Combination of chitosan, collagen, carboxymethyl cellulose in weight ratio of 40:30:30, respectively achieved gradual disintegration of the scaffold in a simulating medium to an open wound after 4 days. This selected scaffold loaded with rosuvastatin revealed increase proliferation of human dermal fibroblasts compared to placebo scaffold. After 30 days of implantation of selected medicated scaffold loaded with/without mesenchymal stem cells and placebo scaffolds to induced wounds in Albino rats, enhanced skin regeneration and absence of scar formation for drug loaded scaffolds were observed. The histopathological study showed the advantage of stem cells-loaded scaffolds through the normal redistribution of collagen in the epidermal layer. In conclusion, rosuvastatin calcium and stem cells loaded in the tested scaffolds proved their potential effect in enhancing skin healing and regeneration.

Design and characterization of emulsified spray dried alginate microparticles as a carrier for the dually acting drug roflumilast.

Roflumilast is a selective inhibitor of phosphodiesterase-4 isoenzyme in lung cells. Having psychiatric adverse reactions when administered orally affects negatively the patients' adherence to the drug. This work aimed to prepare emulsified spray dried alginate microparticles for the pulmonary delivery of roflumilast. Sodium alginate was used as microparticle-forming material, isopropyl myristate as an oil, Tween®80 as surfactant and calcium beta-glycerophosphate as cross-linking agent to enhance the mechanical properties of the particles. The prepared particles were evaluated for their encapsulation efficiency, particle size and in-vitro drug release. From the studied carriers, beta-cyclodextrin (CD) was the best regarding giving formulation with smaller particle size and more sustained drug release. The inhalation profile of CD-based microparticles was investigated using Anderson cascade impactor. The aerosolization profile of CD-based microparticles suggested their efficiency to deliver the drug deep in the lung. The CD-based microparticles possessed more inhibitory effects on the viability of A549 cells and on the pro-inflammatory cytokines (TNF-α, IL-6 and IL-10) compared to the pure drug. Hence, CD-based microparticles could regulate the tumorigenesis besides tumor-associated inflammation. Finally, CD-based microparticles showed more sustained bronchodilatation properties in healthy human volunteers when compared to Ventolin®HFA. CD-based microparticles proved to be a promising carrier for inhaled roflumilast in human.