Transdermal Delivery Of Macromolecules Research Articles

Transdermal Delivery of Macromolecules Using Nano Lipid Carriers.

Skin being the largest external organ, offers an appealing procedure for transdermal drug delivery, so the drug needs to reach above the outermost layer of the skin, i.e., stratum corneum. Small molecular drug entities obeying the Lipinski rule, i.e., drugs having a molecular weight less than 500 Da, high lipophilicity, and optimum polarity, are favored enough to be used on the skin as therapeutics. Skin's barrier properties prevent the transport of macromolecules at pre-determined therapeutic rates. Notable advancements in macromolecules' transdermal delivery have occurred in recent years. Scientists have opted for liposomes, the use of electroporation, low-frequency ultrasound techniques, etc. Some of these have shown better delivery of macromolecules at clinically beneficial rates. These physical technologies involve complex mechanisms, which may irreversibly incur skin damage. Majorly, two types of lipid-based formulations, including Solid Lipid Nanoparticles (SLNs) and Nanostructured Lipid Carriers (NLCs), are widely investigated as transdermal delivery systems. In this review, the concepts, mechanisms, and applications of nanostructured lipid carriers used to transport macromolecules via transdermal routes are thoroughly reviewed and presented along with their clinical perspective.

Transdermal Delivery of Macromolecules Using Two-in-One Nanocomposite Device for Skin Electroporation.

Delivery of hydrophilic molecules through the skin using electroporation is a promising alternative approach to intradermal injection. Recently, we developed a two-in-one electrode/reservoir material composed of carbon nanotubes and agarose hydrogel. In this work, we evaluated the potential of the device to achieve non-invasive transdermal drug delivery using skin electroporation. As it involved an electrode configuration different from the literature, critical questions were raised. First, we demonstrated the efficiency of the device to permeabilize the skin of hairless mice, as observed by propidium iodide (PI) uptake in the nuclei of the epidermis cells through macro fluorescence imaging and histology. Application of Lucifer yellow (LY) at different times after unipolar electroporation treatment demonstrated the partial reversibility of the skin permeabilization after 30 min, and as such, that barrier function properties tended to be restored. We uncovered, for the first time to our knowledge, an intrinsic asymmetry of permeation pathways generated in the stratum corneum during treatment. Electrophoresis was here the main driving force for macromolecule delivery, but it competed with passive diffusion through the generated aqueous pathways for smaller molecules. Finally, we validated 4 kDa dextran labelled with fluorescein isothiocyanate (FD4) as a model molecule to optimize the electrical parameters, needed to improve macromolecule delivery.

Transdermal Penetrating Peptide Conjugated Liposomes as Drug Delivery Carrier Comprising Macromolecules

: The aim of the study was to investigate a system using liposomes and cell penetrating peptides (CPP) for optimal transdermal delivery of macromolecules. Typical DOPE liposomes were prepared with lipid mixture (DOPE:PC:Chol = 1.5:1.5:2.0, molar ratio) and active materials (Rhodamine B; MW ~480 and Dextran-RITC; MW ~10,000). CPP-conjugated DOPE liposomescontaining active materials were prepared by conjugating the peptide to DOPE liposomes (DOPE:PC:Chol:DSPE-PEG-Mal = 1.5:1.1:2.0:0.2 or 0.4, molar ratio). Physical properties of both liposomes were evaluated, including particle size and zeta potential. The particle sizes of typical liposome and CPP-DOPE liposome were approximately 100 nm, and the zeta potential values of both liposomes were approximately -25mV and over +11mV respectively. Moreover, cellular uptake efficiency was assessed by flow cytometry (FACS). CPP-conjugated liposomes resulted higher cellular uptake efficiency compared to typical DOPE liposomes, showing higher fluorescent intensity in CPP-DOPE liposomes. In confocal laser scanning microscope (CLSM) studies, both cellular uptake and skin permeation were visually estimated. In the case of Rhodamine B, having a relatively small molecular weight,absorption into the cell was successful, and showed the highest rate of cellular uptake with CPP-DOPE liposomes. Dextran-RITC, a macromolecule with a relatively bigger molecular weight, showed similar results to Rhodamine B. In terms of skin permeation, CPP-DOPE liposomes containing Rhodamine B showed noticeable skin absorption after 4 and 18 hours, and the permeation range was wider and thicker than that with typical liposomes. For Dextran-RITC, with typical DOPE liposome, it was hardly permeable through the skin, but with CPP-DOPE liposomes, on the other hand, the skin permeations after 4 and 18 hours were remarkable. The improved cellular uptake and skin permeation of the CPP-conjugated liposomes are due to the cationic arginine-rich peptide. In vivo studies also proved that the CPP-conjugated liposomes are superior in depigmentation and anti-wrinkle studies than typical liposomes. These results demonstrate that the CPP-conjugated liposomes could also be effective for transdermal drug delivery of antioxidant and anti-aging therapeutics.

Trypsin as a novel potential absorption enhancer for improving the transdermal delivery of macromolecules

The aim was to assess the effect of trypsin on the transdermal delivery of macromolecules by applying its specific biochemical properties to the stratum corneum of the skin. Fluorescein isothiocyanate (FITC)-labelled dextrans (FDs), with molecular weights of 4 to 250 kDa, and FITC-insulin were used as model macromolecules and a model polypeptide, and the in-vitro transdermal permeation experiments, with or without trypsin (0.1-2.5%), were carried out using rat skin and cultured human epidermis. The mechanism for the enhancement of trypsin was also studied using fluorescence and conventional light microscopy. Trypsin significantly increased the transdermal permeability of all FDs through the rat skin (2.0- to 10.0-fold). It also markedly enhanced the permeation of FD4 through three-dimensional cultured human epidermis (3.1-fold), which was used to evaluate the transport pathways other than the transfollicular route. Furthermore, the permeation flux of FITC-insulin was increased by 10.0-fold with trypsin pretreatment (from 0.02 +/- 0.00 to 0.20 +/- 0.07 microg/cm(2) per h). Mechanistic studies indicated that trypsin affects both the intercellular pathway and the hair follicular route, and may alter stratum corneum protein structures, thereby affecting skin barrier properties. This study suggests that trypsin could be effective as a biochemical enhancer for the transdermal delivery of macromolecules including peptide and protein drugs.

Transdermal delivery of macromolecules by erbium:YAG laser

The aim of this study was to assess the effect of molecular weight (MW) on the transdermal delivery of macromolecules by erbium:yttrium–aluminum–garnet (Er:YAG) laser treatment. Fluorescein isothiocyanate (FITC)-labeled dextran (FD) of increasing MWs (4.4, 19.4, 38, and 77 kDa) was used as the model macromolecules to investigate the skin permeation in vitro. Fluorescence microscopy and scanning electron microscopic (SEM) images were utilized to examine the transport mechanisms of the macromolecules via the skin after laser treatment. The results indicate a significant increase in the permeation of FITC and FD across skin treated by the laser. The MWs of macromolecules and laser fluences were found to play important roles in controlling macromolecular absorption. Transdermal delivery of FD with a MW of at least 77 kDa could be achieved with laser treatment. Follicular routes were significant for FITC permeation, whereas intercellular pathways played important roles on the delivery of FD. Ablation of the stratum corneum (SC) layer, photomechanical stress on intercellular regions, and alterations of the morphology and arrangement of corneocytes are possible mechanisms of how the Er:YAG laser promotes macromolecular delivery. No alteration of viable skin morphology was observed after laser treatment and the partly ablation of the SC may be reversible. Hexameric insulin showed higher skin permeation than did FD with similar MWs (38 kDa) with laser enhancement. From the study presented herein, it is concluded that the Er:YAG laser can be effective for transdermal delivery of macromolecules and hydrophilic permeants such as peptides and protein-based drugs.

Transdermal delivery of macromolecules using skin electroporation.

(1) To evaluate the feasibility of transdermal delivery of macromolecules by skin electroporation. (2) To assess the influence of the molecular weight of the permeant on transport and examine whether there exists a "cut-off" value of molecular weight. (3) To localize the transport pathways of the macromolecules in the skin. FITC-dextran (FD) of increasing molecular weight (4.4, 12 and 38 kDa) were used as model macromolecules to study the extent of transport across hairless rats skin in vitro and to localize their distribution in the skin by confocal scanning laser microscopy. Electroporation enhanced the transport of the macromolecules as compared to passive diffusion. The transdermal delivery by skin electroporation of FITC and FD 4.4 was equivalent whereas transport of higher molecular weight FD was lower but significant. FITC and FD 38 were observed in the epidermis both around and in the keratinocytes. Transdermal and topical delivery of macromolecules of at least 40 kDa can be achieved by skin electroporation.

Reversible Skin Permeabilization for Transdermal Delivery of Macromolecules

Great advances in transdermal delivery of macromolecules have been made within the last few years using ultradeformable liposomes, electroporation, and low-frequency ultrasound, each of which has been shown to deliver macromolecules at clinically useful rates. Until recently, delivery of high-molecular-weight compounds across the skin was not a realistic option, since the skin's great barrier properties prevent transport of macromolecules across human skin at therapeutically relevant rates. An overview of chemical (liposomes and chemical enhancers) and physical (iontophoresis, electroporation, and ultrasound) methods of enhancement is presented, with emphasis on work published within the last two years. Key experimental results and their mechanistic interpretation are provided for each enhancement technique.