Influence of cyclodextrins on the proliferation of HaCaT keratinocytes in vitro

Abstract

Safety and efficacy of pharmaceutical agents can be greatly improved by encapsulation within, or covalent attachment to, a biomaterial carrier. Drug delivery systems must deliver the necessary amount of drug to the targeted site for a necessary period of time, both efficiently and precisely. Various kinds of high-performance biomaterials are being constantly developed for this purpose. Cyclodextrins are potential candidates for such a role, because of their ability to alter physical, chemical, and biological properties of guest molecules through the formation of inclusion complexes. The alpha-, beta-, and gamma-cyclodextrins are widely used natural cyclodextrins, consisting of six, seven, and eight D-glucopyranose residues, respectively, linked by -1,4 glycosidic bonds into a macro cycle. Each cyclodextrin has its own ability to form inclusion complexes with specific guests, an ability, which depends on a proper fit of the guest molecule into the hydrophobic cyclodextrin cavity. The most common pharmaceutical application of cyclodextrins is to enhance the solubility, stability, and bioavailability of drug molecules. Such kinds of ligand-receptor complexes can be used for different applications, e.g., for a transdermal therapeutic system (TTS) or in biofunctional textiles. The aim of this study was the investigation of the influence of the different cyclodextrins on the cell proliferation using HaCaT keratinocytes as an in vitro test system. Moreover, the study was performed to find harmless and nontoxic cyclodextrin concentrations for dermal applications. By means of different independent in vitro tests could be confirmed that alpha-, beta-, and gamma-cyclodextrins in concentrations up to 0.1% (w/v) do not show any antiproliferative influence on HaCaT keratinocytes. Sometimes even proliferative effects could be found. However, all used cyclodextrins (besides gamma-cyclodextrin and its derivatives) in concentrations of 0.5 and 1% (w/v), respectively, exert a cytotoxic influence on the proliferation of HaCaT keratinocytes. On the basis of these findings, the following rank order of cyclodextrins regarding their cytotoxicity was proposed: M-beta-CD > beta-CD > HP-beta-CD > alpha-CD > (gamma-CD). It could be confirmed that beta-CD and M-beta-CD trigger the activity of the effectors caspases -3 and -7. A significant increase of LDH release could be found for beta-CD and methyl-beta-CD in concentrations of 0.5 and 1% (w/v). The calculated cytotoxicity amounted 45 and 79%, respectively. The measurements of the interleukins IL-6 and IL-8 confirmed the findings of the proliferation assays as well as the LDH measurements. These findings may provide further rationale to the use of CDs in topical formulations for dermal and transdermal drug delivery or as raw material to functionalize textiles for medical applications.

Evaluation of Biocompatibility and Cytotoxicity Using Keratinocyte and Fibroblast Cultures

“…7 ). These results confirmed the negative effect of high concentrations of ␤ -cyclodextrin as found in the proliferation assay as well as the LDH measurements for the biocompatibility study on ␤ -cyclodextrin [10] .…”

“…This method is quick and easily adaptable for different types of cells and test conditions, but the assay is sensitive to interference from compounds with inherent LDH activity like serum-supplemented culture media. Using ATP and LDH measurement, our group has been able to further explore the cytotoxic influence of ␤ -cyclodextrin in concentrations of 0.5 and 1% (w/v) on HaCaT cells [10] . A significant increase in LDH release was found that accompanied the rapid decline of cell viability ( fig.…”

“…Cyclodextrins are cyclic oligosaccharides that are able to form inclusion complexes with a variety of substances. Hence, they are potential candidates for drug delivery systems because of their ability to alter physical, chemical and biological properties of their guest molecules [10] .…”

Protein encapsulation in polymeric microneedles by photolithography

“…61 In our study, low cell toxicities were observed in all the three cell lines for cells treated with polymer extracts. The percentage cytotoxicity for cells treated from 24-72 hours did not vary significantly (P .…”

Introduction of phospholipids to cultured cells with cyclodextrin

“…Previously, exposure of HaCaT cells to me ␤ -CD was shown to cause i ) a leakage of lactate dehydrogenase, ii ) an increase in caspase-3/7 activity, iii ) a decrease of ROS concentration, and iv ) an increase in interleukin 6 and 8 ( 35 ). In addition, inhibitory effects, such as inhibition of cell growth and cell cycle arrest, have been reported for various cell types ( 36,37 ).…”