In-vitro evaluation of dexpanthenol-loaded nanofiber mats for wound healing

Purpose: To prepare a novel dexpanthenol (DEX)-loaded nanofiber mats for wound healing.Methods: A novel bioactive wound dressing formulation with  dexpanthenol was developed by electrospinning method. Poly (lactic-co-glycolic acid), poly (ethylene oxide) and poly (caprolactone) were used as polymers. Morphological features, swelling properties, in-vitro release behavior, and cell viability properties of the formulations were evaluated.Results: Morphological examination of mats confirmed successful formation of the fibers. Swelling of nanofiber mats results was 34.44 ± 1.05, 18.59 ± 2.11, 86.06 ± 3.25 and 44.62 ± 1.75 % for polycaprolactone (PCL), PCL + DEX, poly lactic-co-glycolic acid (PLGA) and PLGA + DEX, respectively, and occurred in a controlled manner. PLGA + DEX formulation has advantage over PCL + DEX and poly (ethylene oxide) (PEO) + DEX due to controlled in-vitro release of DEX. The highest cell viability was afforded by PLGA+DEX formulation.Conclusion: DEX-loaded PLGA nanofiber formulation may be useful as an alternative wound dressing due its suitable mechanical and biological properties.Keywords: Dexpanthenol, Electrospinning, Wound dressing,  Polycaprolactone, Poly lactic-co-glycolic acid, Poly (ethylene oxide

Transdermal olarak uygulanacak ilaç yüklü formülasyonlar için yenilikçi üretim yöntemlerinin tasarımı ve etkinliklerinin incelenmesi

Transdermal ilaç taşıyıcı sistemler, etkin maddenin deri aracılığıyla sistemik dolaşıma geçişini sağlayan ilaç uygulama yöntemlerindendir. Transdermal ilaç taşıyıcı sistemlerden olan yamalar ve mikro iğnelerin üretim tekniklerinde yenilikçi yaklaşımların geliştirildiği çalışmalarda etkin madde olarak montelukast sodyum (MS) kullanılmıştır. Üç boyutlu (3B) yazıcı filamentleri sıcak eriyik ekstrüzyon (SEE) yöntemi ile üretilmiştir. Filamentlerin mekanik dayanıklılıkları incelenmiş ardından 3B yazıcı ile yamaların üretiminde kullanılmıştır. MS ile ilgili karakterizasyon çalışmaları gerçekleştirilmiştir. MS için yüksek performanslı sıvı kromatografisi ile miktar tayini yöntemi geliştirilip valide edilmiştir. Zorlayıcı testlerde MS'nin ısı ve ışık maruziyetine karşı dayanıklılığı ölçülmüştür. FT-IR, DTK, TGA ve UV spektrumu analizlerinin yanı sıra çözünürlük, lipit/su partisyon katsayısı tayinleri de gerçekleştirilmiştir. Karakterizasyon çalışmalarının ardından MS yüklü yamalar 3B yazıcı ile üretilip, mekanik testler ve in vitro salım çalışması yapılmıştır. 3B yazıcılarda sıklıkla kullanılan poli(laktik asit) filamentler ve bu filamentlerle hazırlanan yamalar kıyaslama amacı ile kullanılmıştır. Sadece MS'den oluşan mikro iğneler kalıba döküm yöntemi ile üretilmiş ve maksimum yükleme kapasitesine ulaşılmıştır. Mikro iğneler ile görüntüleme çalışmalarını mekanik analizler izlemiştir. In vitro ve ex vivo salım çalışmaları yapılmıştır. Stabilite çalışmalarında MS miktarı ve mekanik özellikler incelenmiştir. Deri modelleri ile in vitro transdermal geçiş analizleri yapılmış, mikro iğnelerin keratinosit tabakasını aşabilme yeteneği incelenmiştir. MS'nin ve mikro iğnelerin biyouyumluluğunun incelenmesi için in vitro sitotoksisite ve irritasyon çalışmaları yapılmıştır. Mikro iğne uygulamasının ardından in vitro doku yüzeyi morfolojisi taramalı elektron mikroskobu ve model histolojisi için hematoksilen ve eozin boyama yöntemleri ile incelenmiştir. Çalışmalarımızın sonucunda sadece oral yol ile uygulanabilen farmasötik formları olan MS için yama ve mikro iğneler geliştirilmiştir. Transdermal uygulama ile MS'nin ilk geçiş etkisinden korunacağı ve biyoyararlanımın artacağı ayrıca oral yoldan ilaç alamayan hastalar için alternatif bir uygulama yolu oluşturacağı düşünülmektedir. Geliştirilen üretim yöntemleri ile farklı etkin maddeler de kullanılarak hastaların yaşam kalitesini artırmaya yönelik çeşitli transdermal ilaç taşıyıcı sistemlerin geliştirilebileceği öngörülmektedirTransdermal drug delivery systems are one of the drug delivery systems, enabling the transport of the drug to the systemic circulation through the skin. Montelukast sodium (MS), was used as an active ingredient in our studies, which includes innovative approaches have been developed for the production techniques of patches and microneedles as transdermal drug delivery systems. Three-dimensional (3D) printer filaments were produced with hot melt extrusion (HME) method. The mechanical strengths of the filaments were investigated and used with the 3D printer for fabrication of patches. Characterization studies of MS was carried out. Quantitative determination of MS by high performance liquid chromatography has been developed and validated. As the stress tests, the stability of MS with heat and light exposure was investigated. In addition to the FT-IR, DSC, TGA and UV spectrum analyzes, solubility, lipid/water partition coefficient analyzes were also carried out. Following the characterization studies, MS loaded patches were produced with a 3D printer and mechanical tests and in vitro release studies were performed. Filaments made of poly(lactic acid) which are frequently used in 3D printers and the patches prepared by using these filaments were used for comparison. Microneedles made of pure MS were fabricated with molding method and maximum loading capacity was achieved. Imaging studies of microneedles were followed by mechanical analyzes. In vitro and ex vivo release studies were carried out. Stability of microneedles were examined in terms of MS amount and mechanical properties. In vitro transdermal permeation studies and ability of perforation of keratinocyte layer of microneedles were carried out with skin models. In vitro cytotoxicity and irritation studies were realized to investigate the biocompatibility of MS and microneedles. Morphology of the surface of the tissue was investigated with scanning electron microscope and histology studies were conducted with hematoxylin eosin staining methods. As the result of our studies, patches and microneedles have been developed for MS, which has only orally administered pharmaceutical forms. By transdermal administration of MS, the first pass effect would be avoided and the bioavailability would be increased additionally an alternative route of administration to the oral route may be constituted for the patients who cannot use oral route. It is envisaged that developed production methods, various transdermal drug delivery systems can be used with different active ingredients for improving the quality of patients

Single and multi-dose drug loaded electrospun fiber mats for wound healing applications

The unique properties of electrospun fibers have received significant attention from the scientific community as suitable candidates for biomedical engineering and drug delivery applications. Their morphological similarity to the extracellular matrix fits the purpose of wound healing by providing a scaffold for cell proliferation while acting as a drug delivery system. One challenge to this drug delivery system's commercialization is the electrospun fiber mat consistency. Reproducibility and accuracy, as well as a large production volume to meet the market demand, should be handled effectively. In this study, we addressed this challenge by optimizing the fabrication method for single and multi-dose production by using flat and drum collectors, respectively. Polycaprolactone was used as a biodegradable polymer to produce fiber structure, and melatonin was used to promote wound healing. Melatonin-loaded polycaprolactone fiber mats were successfully fabricated with RSD values of 0.82% (diameter) and %1.94% (width) using flat and drum collectors, respectively. In addition, functionalized polycaprolactone fiber mats were prepared using peptide fragments, named FnIII9*10, that have both Synergy and RGD sides of fibronectin, with a binding efficiency of 53.9% to the blank PCL fiber mats. The proliferative dose of melatonin was found as 125 mu M and 250 mu M for MRC-5 lung fibroblast cells. The biocompatibility of the fiber mats was studied with BJ skin fibroblast, and HACAT immortalized human keratinocytes. Melatonin-loaded polycaprolactone fiber mats showed increased cell viability values of 178% and 141% for both cell lines. The presented study shows the potential of the commercialization of electrospun fiber mats and gets us one step closer to seeing new-generation wound healing products in the pharmaceutical industry.Scientific and Technological Research Council of Turkey-T?; TAK [117S213]This work was supported by The Scientific and Technological Research Council of Turkey-T?BI ? TAK (Project number: 117S213) . The authors would like to thank Prof. Dr.Petek Ballar Kirmizibayrak for her support in providing the MRC-5 lung fibroblast cell line and cell culture lab facilities