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Analysis of In Vitro Cytotoxicity of Carbohydrate-Based Materials Used for Dissolvable Microneedle Arrays

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Abstract

Purpose

Dissolvable microneedle arrays (MNAs) can be used to realize enhanced transdermal and intradermal drug delivery. Dissolvable MNAs are fabricated from biocompatible and water-soluble base polymers, and the biocargo to be delivered is integrated with the base polymer when forming the MNAs. The base polymer is selected to provide mechanical strength, desired dissolution characteristics, and compatibility with the biocargo. However, to satisfy regulatory requirements and be utilized in clinical applications, cytotoxicity of the base polymers should also be thoroughly characterized. This study systematically investigated the cytotoxicity of several important carbohydrate-based base polymers used for production of MNAs, including carboxymethyl cellulose (CMC), maltodextrin (MD), trehalose (Treh), glucose (Gluc), and hyaluronic acid (HA).

Methods

Each material was evaluated using in vitro cell-culture methods on relevant mouse and human cells, including MPEK-BL6 mouse keratinocytes, NIH-3T3 mouse fibroblasts, HaCaT human keratinocytes, and NHDF human fibroblasts. A common laboratory cell line, human embryonic kidney cells HEK-293, was also used to allow comparisons to various cytotoxicity studies in the literature. Dissolvable MNA materials were evaluated at concentrations ranging from 3 mg/mL to 80 mg/mL.

Results

Qualitative and quantitative analyses of cytotoxicity were performed using optical microscopy, confocal fluorescence microscopy, and flow cytometry-based assays for cell morphology, viability, necrosis and apoptosis. Results from different methods consistently demonstrated negligible in vitro cytotoxicity of carboxymethyl cellulose, maltodextrin, trehalose and hyaluronic acid. Glucose was observed to be toxic to cells at concentrations higher than 50 mg/mL.

Conclusions

It is concluded that CMC, MD, Treh, HA, and glucose (at low concentrations) do not pose challenges in terms of cytotoxicity, and thus, are good candidates as MNA materials for creating clinically-relevant and well-tolerated biodissolvable MNAs.

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Data Availability

The authors declare that all data supporting the findings of this study are available within the paper and its supplementary material, source data for the figures in this study are available from the authors upon request.

Abbreviations

CMC:

Carboxymethyl cellulose

DMEM:

Dulbecco’s Modified Eagle’s Medium

FBS:

Fetal bovine serum

FDA:

U.S. Food and Drug Administration

Gluc:

Glucose

GMP:

Good manufacturing practices

HA:

Hyaluronic acid

ISO:

International Organization for Standardization

MD:

Maltodextrin

MNA:

Microneedle array

PBS:

Phosphate-buffered saline

Treh:

Trehalose

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Authors and Affiliations

  1. Department of Mechanical Engineering, Carnegie Mellon University, Pittsburgh, Pennsylvania, 15213, USA

    Ezgi P. Yalcintas, Emrullah Korkmaz & O. Burak Ozdoganlar

  2. Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, Pennsylvania, 15213, USA

    Daniel S. Ackerman, Jonathan W. Jarvik & Marcel P. Bruchez

  3. Molecular Biosensor and Imaging Center, Carnegie Mellon University, Pittsburgh, Pennsylvania, 15213, USA

    Daniel S. Ackerman, Cheryl A. Telmer, Jonathan W. Jarvik, Phil G. Campbell & Marcel P. Bruchez

  4. Department of Biomedical Engineering, Carnegie Mellon University, Pittsburgh, Pennsylvania, 15213, USA

    Phil G. Campbell & O. Burak Ozdoganlar

  5. Department of Chemistry, Carnegie Mellon University, Pittsburgh, Pennsylvania, 15213, USA

    Marcel P. Bruchez

  6. Department of Materials Science and Engineering, Carnegie Mellon University, Pittsburgh, Pennsylvania, 15213, USA

    O. Burak Ozdoganlar

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  1. Ezgi P. Yalcintas
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Contributions

Conceptualization, E.P.Y, E.K., P.G.C., and O.B.O.; Methodology, E.P.Y., E.K., D.S.A., P.G.C., and M.P.B.; Investigation, E.P.Y., D.S.A., E.K., and P.G.C.; Resources, O.B.O., P.G.C., M.P.B., and J.W.J.; Writing - original draft, E.P.Y. and E.K.; Writing - review editing, E.P.Y., E.K., P.G.C., M.P.B., J.W.J., C.A.T., and O.B.O.; Visualization, E.P.Y., E.K., and D.S.A.; Supervision, E.K., P.G.C., M.P.B., J.W.J., and O.B.O.; Project administration, P.G.C., O.B.O., and M.P.B.; Funding acquisition; O.B.O., P.G.C., M.P.B., and J.W.J.

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Correspondence to O. Burak Ozdoganlar.

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Yalcintas, E.P., Ackerman, D.S., Korkmaz, E. et al. Analysis of In Vitro Cytotoxicity of Carbohydrate-Based Materials Used for Dissolvable Microneedle Arrays. Pharm Res 37, 33 (2020). https://doi.org/10.1007/s11095-019-2748-7

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