In Vivo Wound-Healing Efficacy of Insulin-Loaded Strontium-Cross-Linked Alginate-Based Hydrogels in Diabetic Rats.

The wound-healing effect of insulin is well studied and reported. However, prolonged topical application of insulin without compromising its biological activity is still a challenge. In this study, the effect of topically delivered insulin on promoting wound healing in diabetic animals was evaluated. Alginate diamine PEG-g-poly(PEGMA) (ADPM2S2) was the material used for the topical delivery of insulin. ADPM2S2 hydrogels release insulin and strontium ions, and they synergistically act to regulate different phases of wound healing. Insulin was released from the ADPM2S2 hydrogel for a period of 48 h, maintaining its structural stability and biological activity. In vitro studies were performed under high-glucose conditions to evaluate the wound-healing potential of insulin. Insulin-loaded ADPM2S2 hydrogels showed significant improvement in cell migration, proliferation, and collagen deposition, compared to control cells under high-glucose conditions. Immunostaining studies in L929 cells showed a reduction in phospho Akt expression under high-glucose conditions, and in the presence of insulin, the expression increased. The gene expression studies revealed that insulin plays an important role in regulating the inflammatory phase and macrophage polarization, which favors accelerated wound closure. In vivo experiments in diabetic rat excision wounds treated with insulin-loaded ADPM2S2 showed 95% wound closure within 14 days compared with 82% in control groups. Thus, both the in vitro and in vivo results signify the therapeutic potential of topically delivered insulin in wound management under high-glucose conditions.

Design and evaluation of propranolol hydrochloride loaded thiolated Zein/PEO electrospun fibrous matrix for transmucosal drug delivery.

Thiolated polymers have garnered wide attention from researchers on mucoadhesive drug delivery. This work explores the thiolation of zein protein using cysteine amino acid via the EDC crosslinker. The optimization of thiolation and purification have been done and confirmed using Ellman's assay and Raman spectra. The thiolated Zein/PEO polymer blend has been appraised for electrospinning to fabricate fibrous matrices. The extent of thiol modification augmented the mechanical properties and adhesion in rabbit intestinal mucosa. In vitro cytotoxicity evaluations such as direct contact assay, MTT assay, and live dead assay performed in RPMI 2650 cells corroborated the non-cytotoxicity of the fabricated matrices with and without propranolol hydrochloride (PL). Detailed drug release studies were conducted in PBS. Drug release in PBS followed the Korsmeyer Peppas model of release. On treating RPMI 2650 cells with the matrix, F-actin and adherens junctional proteins retained integrity, and consequently, drug permeation would proceed through the transcellular transport mechanism. Transepithelial electrical resistance (TEER) measurement of the RPMI 2650 cell monolayer also supported the transcellular transport mechanism. Ex vivo permeation study through porcine buccal mucosa showed 41.26 ± 0.56% PL permeation within 24 h of study. It validated the competence of the electrospun thiolated Zein/PEO matrix for transmucosal drug delivery.

Electrospun Mucoadhesive Zein/PVP Fibroporous Membrane for Transepithelial Delivery of Propranolol Hydrochloride.

Mucoadhesive drug delivery systems have been extensively studied to effectively reduce the limitations of conventional drug delivery systems. Zein and polyvinyl pyrrolidone (PVP) are appraised for mucoadhesive properties. This study focuses on developing a mechanically stable zein/PVP electrospun membrane for propranolol hydrochloride (PL) transport. Fourier transform infrared, Raman spectra, and swelling studies gave evidence for PVP crosslinking, whereas circular dichroism spectroscopy revealed crosslinking of zein owing to the conformational change from α-helix to ß-sheet. A 10 h thermal treatment of zein/PVP imparted 3.92 ± 0.13 MPa tensile strength to the matrix. Thermally crosslinked electrospun zein/PVP matrix showed 22.1 ± 0.1 g mm work of adhesion in porcine buccal mucosa tissue. Qualitative and quantitative evaluation of cytotoxicity in RPMI 2650 has been carried out. The in vitro drug release profile of PL from thermally crosslinked zein/PVP best fitted with the Korsmeyer-Peppas model. Immunostaining of ß-catenin adherens junctional protein confirmed the absence of paracellular transport through the junctional opening. Still, drug permeation was observed through the porcine buccal mucosa, attributed to the transcellular transport of PL owing to its lipophilicity. The ex vivo permeation of PL through porcine buccal mucosa was also evaluated.

UV-Crosslinked Electrospun Zein/PEO Fibroporous Membranes for Wound Dressing.

Electrospun zein membranes are suitable for various biomedical applications. A UV-crosslinked electrospun membrane of a zein/PEO blend for wound healing application was explored in this work. The improvement in mechanical properties of the membrane after UV crosslinking was attributed to the change in protein conformation from an α-helix to a ß-sheet. The circular dichroism (CD) spectra and FTIR spectra confirmed this conformational change. XRD analysis was shown to prove the amorphous nature of polymer blends with specific broad peaks at 2θ = 9° and 20°. The water vapor transmission rate (WVTR) of the membrane was found to be in the range of 1500-2000 g m-2 day-1, which was well suited with that of commercially available wound dressing material. Enough number of available functional groups like thiol, amino, and hydroxyl groups supplement a blood clotting index (BCI) to the matrix, causing 99% BCI within 4 min. A 91% cell viability result in the MTT assay with human dermal fibroblast cells confirmed the noncytotoxicity of the membrane. Tripeptides produced after the thermolysin-based hydrolysis of zein caused inhibition of TGF ß1 expression and thus increased fibroblast and collagen production. The membrane stimulated 54% more collagen production compared to control cells at day 2 and caused 84% wound closure in human dermal fibroblast cells, which were desirable index markers of a potential wound care material.

Elastin-like recombinamers with acquired functionalities for gene-delivery applications.

In this work, well-defined elastin-like recombinamers (ELRs) were studied as a choice to the existing nonviral vectors due to their biocompatibility and ease of scale-up. Functional motifs, namely penetratin and LAEL fusogenic peptides were incorporated into a basic ELR sequence, and imidazole groups were subsequently covalently bound obtaining ELRs with new functionalities. Stable polyplexes composed of plasmid DNA and ELRs were formed. A particle size around 200 nm and a zeta potential up to nearly +24 mV made them suitable for gene delivery purposes. Additionally, viability and transfection assays with C6 rat glioma cell line showed an increase in the cellular uptake and transfection levels for the construction containing the LAEL motif. This study highlights the importance of controlling the polymer functionality using recombinant techniques and establishes the utility of ELRs as biocompatible nonviral systems for gene-therapy applications.

Recent advances in the oral delivery of insulin.

Insulin was discovered over 90 years ago. However oral insulin still remains a challenging and elusive goal. Extensive efforts are being made worldwide for developing noninvasive drug delivery systems, mainly via oral route as it is the most widely accepted means of administration. The main barriers faced in oral protein delivery are the enzymatic degradation and poor permeability across the intestinal wall. The approaches for developing an oral insulin delivery system mainly focus on overcoming these barriers. To overcome the gastro-intestinal barriers various types of formulations such as insulin conjugates, permeation enhancers, micro/nanoparticles, liposomes etc. are investigated. In the recent years a number of advances have taken place in understanding the needs and workable mechanisms of improved oral peptide delivery systems. In this review the recent patents on oral insulin is focused. Emphasis is on the technologies based on permeation enhancers and nanoparticle based carrier systems.

Challenges and advances in nanoparticle-based oral insulin delivery.

For many decades, rigorous efforts have been made worldwide to develop a successful oral insulin-delivery system, which still remains an elusive goal. However, over the past few years, tremendous understanding has evolved in the development of biocompatible and biodegradable polymers, synthesis of nanopeptide delivery systems, biocompatibility and its cellular uptake mechanisms. With these advancements, efforts are being directed toward nanoparticle-based oral peptide-delivery systems. It is established that nanoparticles enhance oral bioavailability by facilitating insulin uptake via a transcellular or paracellular pathway. In this process, the particle also reaches the systemic circulation. Hence, biocompatibility and the half-life of the particles in the systemic circulation is an important aspect that needs to be looked into. In this review, the various approaches adopted for nanoparticle-based oral insulin delivery, uptake mechanisms, biocompatibility and bioavailability of the nanoparticle are discussed.