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Generation of Genetically Modified Organotypic Skin Cultures Using Devitalized Human Dermis
Published on: December 14, 2015
Epidermal and dermal cell-composed organospheres to assess microplastic-induced skin toxicity
Yunxia Hu1, George Michael Nicolas2, Yongde Cai3
1Institute of Biopharmaceutical and Health Engineering (iBHE), Tsinghua Shenzhen International Graduate School (SIGS), Tsinghua University, Shenzhen, 518055, China; Key Lab of Industrial Biocatalysis, Ministry of Education, Shenzhen, 518055, China; Key Lab of Active Proteins and Peptides Green Biomanufacturing of Guangdong Higher Education Institutes, Tsinghua Shenzhen International Graduate School, Shenzhen, 518055, China.
Abstract:
The alarming prevalence of microplastics (MPs) in personal care products and their inherent presence in animals and humans necessitate the development of alternative experimental approaches to evaluate their cytotoxicity, mechanisms, and long-term effects. Microfluidic bead-jet printed skin organospheres, comprising epidermal and dermal cells, present significant promise in addressing MP research, creating high-throughput and reproducible screening systems. Medium-density bioprinted E18 pregnant mouse fetus epidermal and dermal cell (Epi-Dc) organospheres exhibited fast self-aggregation, migration, and cellular differentiation. Co-culture revealed a size-dependent MP uptake rate, with smaller MPs having higher internalization ratios. The 100 nm MP penetrated the epidermal layer and migrated toward the central region, while the 500 nm MP remained in the peripheral area. Ultraviolet A (UVA)-irradiated MPs also significantly increased oxidative stress, cell apoptosis, and ROS levels via upregulation of oxidative stress-associated genes (SOD, p53, and Bax). Furthermore, vitamin C treatment following MP-UVA irradiation decreased reactive oxygen species levels, while cell viability assay using conventional skin-associated drugs indicate dose- and time-dependent induced cytotoxicity. These results demonstrate the broad applications of Epi-Dc organospheres in skin toxicity evaluation and rapid drug testing.
Insights
New bioprinted skin organospheres offer a promising alternative for testing microplastic (MP) toxicity. These models show size-dependent MP uptake and reveal UVA-irradiated MPs induce significant oxidative stress and apoptosis.
Area of Science:
- Biomedical Engineering
- Toxicology
- Materials Science
Background:
- Microplastics (MPs) are increasingly prevalent in consumer products and the environment, posing risks to human health.
- Current methods for evaluating MP cytotoxicity are limited, necessitating novel experimental approaches.
- Skin organospheres offer a potential in vitro model for assessing MP effects on human skin.
Purpose of the Study:
- To develop and validate microfluidic bead-jet printed skin organospheres for microplastic toxicity assessment.
- To investigate the size-dependent uptake and effects of microplastics on skin cells.
- To evaluate the impact of UVA-irradiated microplastics on oxidative stress and apoptosis.
Main Methods:
- Bioprinting of epidermal and dermal cells (Epi-Dc) from E18 mouse fetuses into organospheres using microfluidic bead-jet technology.
- Co-culture of organospheres with varying sizes of microplastics (100 nm and 500 nm).
- Assessment of microplastic uptake, cellular differentiation, oxidative stress markers (ROS), apoptosis, and gene expression (SOD, p53, Bax).
- Evaluation of the protective effects of Vitamin C and cytotoxicity of conventional drugs.
Main Results:
- Bioprinted Epi-Dc organospheres demonstrated rapid self-aggregation, migration, and differentiation.
- A size-dependent microplastic uptake was observed, with smaller MPs showing higher internalization.
- 100 nm MPs penetrated the epidermal layer, while 500 nm MPs remained peripheral.
- UVA-irradiated MPs significantly elevated oxidative stress, apoptosis, and ROS levels.
- Vitamin C mitigated ROS levels, and drug assays showed dose- and time-dependent cytotoxicity.
Conclusions:
- Epi-Dc organospheres are a viable high-throughput screening system for evaluating microplastic skin toxicity.
- The model effectively demonstrates size-dependent microplastic uptake and UV-induced damage.
- This bioprinting approach facilitates rapid drug testing and toxicological assessments for skin-related applications.
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