A study on the role of eugenol encapsulated liposomes in facilitating neuron -microglia mediated wound recovery

Revi, N. and Sankaranarayanan, S.A. and Rengan, Aravind Kumar (2022) A study on the role of eugenol encapsulated liposomes in facilitating neuron -microglia mediated wound recovery. Materialia, 23 (101454). pp. 1-11. ISSN 2589-1529

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Abstract

Activated microglia helps in resolving injury and supports wound healing. However, prolonged activation of microglia can lead to the aggravation of wound. Objective: The present study investigates neuronal injury-induced inflammatory response in microglia and how it is influenced by the treatment with an anti-inflammatory formulation, Eugenol encapsulated liposomes. Method: We introduced injury in an in vitro model of neuronal and microglia origin cell lines using scratch assay and evaluated wound closure rates with respect to treatment. On similar lines, both cell lines were treated in a combinatorial model using Eugenol encapsulated liposomes and investigated the effect of secreted factors in Conditioned Media (CM) on Reactive Oxygen Species (ROS) production, cell viability etc. We also evaluated the polarization properties of microglia in case of an injury by evaluating the amount of nitrite release. Further, in a co-culture model, we introduced neuronal injury in the vicinity of microglia. We then evaluated the inflammatory response of microglia after incubating the injured cells with the formulation for an hour. Results: Individual wounds, CM and co-culture models of neuronal injury treated with Eugenol encapsulated liposomes indicated a decrease in the production of ROS and pro-inflammatory microglia, with the latter displaying an enhanced difference (50–75% with respect to control). The markers produced by microglia cells with respect to an inflammatory condition followed by treatment with Eugenol encapsulated liposomes were investigated using ELISA and Western blotting. There was a reduction in the amount of pro-inflammatory markers TNF-α, IL-1β and an increase in the amount of anti-inflammatory markers IL-10 and IL-4. In control and treatment groups, cardiac rhythm was found to be in the basal levels (100–150 beats per minute). Viability of the embryos were also unaffected due to the treatment in individual group of 20 embryos each. Further, uptake of the formulation in embryos 2,3 & 4 days post fertilization (dpf) were analyzed. Co-encapsulating tracker dye, Nile red within the formulation indicated encapsulated tracker dye crossing organ barriers with respect to free dye. Conclusion: Neuronal injury causes microglial activation. Prolonged microglial activation hinders wound closure and can lead to secondary trauma. The present study was conducted using an in vitro scratch assay wound model. Treating the site of injury with an anti-inflammatory formulation, Eugenol loaded liposomes, converts pro-inflammatory microglia to an anti-inflammatory state and increases the rate of wound closure. © 2022

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IITH Creators:

IITH Creators	ORCiD
Rengan, Aravind Kumar	https://orcid.org/0000-0003-3994-6760

Item Type:

Article

Additional Information:

The authors would like to acknowledge Ministry of Edu- cation (MoE), Dept. of Biotechnology (DBT), Dept. of Science & Technology (DST), Govt. of India for the financial support through DST-Inspire (DST/INSPIRE/04/2015/000377), DBT-NNT (BT/NNT/28/1386/2017), DST-AMT (DST/TDT/AMT/2017/227), MHRD IMPRINT (4291), ICMR grant No35/1/2020-GIA/Nano/BMS and SERB-CRG (CRG/2020/005069).

Uncontrolled Keywords:

Eugenol, Microglia, Neuroinflammation, Neurons, Traumatic brain injury

Subjects:

Others > Biotechnology
Biomedical Engineering

Divisions:

Department of Biomedical Engineering

Depositing User:

. LibTrainee 2021

Date Deposited:

23 Jun 2022 11:30

Last Modified:

30 Jun 2022 07:33