Co-cultivation of PBMCs microvesicle activated by MSCS and OS-OCs

J. Pharm. Pharmacogn. Res., vol. 10, no. 5, pp. 782-790, September-October 2022.

Original Article

Activation of microvesicle peripheral blood mononuclear cells by mesenchymal stem cells secretome co-cultivated with osteosarcoma stem cell

[Activación de microvesículas de células mononucleares de sangre periférica por secretoma de células madre mesenquimales cocultivadas con células madre de osteosarcoma]

Fachrizal Arfani Prawiragara1,2, Ferdiansyah2*, Mouli Edward2, Dwikora Novembri Utomo2, Mohammad Hardian Basuki2, Alexander Patera Nugraha3, Fedik Abdul Rantam4

1Magister Clinical Medicine Program, Orthopedic and Traumatology Department, General Academic Dr. Soetomo Hospital/Faculty of Medicine, Airlangga, Surabaya, Indonesia.

2Department of Orthopaedic and Traumatology, General Academic Dr. Soetomo Hospital/Teaching Hospital, Faculty of Medicine, Airlangga University, Surabaya, Indonesia.

3Department of Orthodontics, Faculty of Dental Medicine, Airlangga University, Surabaya, Indonesia.

4Stem Cell Research Center and Development, Airlangga University, Surabaya, Indonesia.

*E-mail: ferdiansyah@fk.unair.ac.id

Abstract

Context: Microvesicle is a cell micro molecule that may play a role in the process of osteosarcoma stem cell apoptosis.

Aims: To investigate the activity of peripheral blood mononuclear cells (PBMCs) through the secretion of interleukin-2 (IL-2), interleukin-6 (IL-6), interleukin-10 (IL-10), C-X-C Motif Chemokine Ligand 13 (CXCL13) and tissue inhibitor of metalloproteinases-3 (TIMP-3) on co-cultivation of peripheral blood mononuclear cells (PBMCs) sensitized by mesenchymal stem cell secretome (MSCS) co-cultivated with osteosarcoma stem cells (OS-SCs).

Methods: This study was true experimental with a post-test only control group design. This was in vitro study PBMSCs sensitized by MSCS as then samples were divided into 4 treatment groups, respectively: Zero-day treatment (P0) PBMCs were co-cultivated with OS-SCs for 0 hours; First treatment (P1) PBMCs were co-cultivated with OS-SCs for 1 hour; Second treatment (P2) PBMCs were co-cultivated with OS-SCs for 2 days; Third treatment (P3) PBMCs were co-cultivated with OS-SCs for 4 days. The examination method used in this study was flow cytometry and indirect enzyme-linked immunosorbent assay (ELISA). The data were statistically analyzed with analysis of variance (ANOVA) with a p≤0.05 considered a significant difference.

Results: There was a tendency for a significant increase in extravesicular secretion in the secretion of IL-2, IL-6, IL-10, CXCL13, TIMP3, in the microvesicle PBMCs when sensitized by MSCs secretome co-cultivated with OS-SCs environment co-cultivated after the fourth day with significantly different between groups (p≤0.05).

Conclusions: PBMSCs’ microvesicle such as IL-2, IL-6, IL-10, CXCL13, TIMP3 was significantly sensitized by MSCS and co-cultivated with OS-SCs after the fourth day of in vitro.

Keywords: medicine; non-communicable disease; non-infectious disease; osteosarcoma; stem cells.

Resumen

Contexto: La microvesícula es una micromolécula celular que puede desempeñar un papel en el proceso de apoptosis de las células madre del osteosarcoma.

Objetivos: Investigar la actividad de las células mononucleares de sangre periférica (PBMC) a través de la secreción de interleucina-2 (IL-2), interleucina-6 (IL-6), interleucina-10 (IL-10), C-X-C Motif Chemokine Ligand 13 (CXCL13) e inhibidor tisular de metaloproteinasas-3 (TIMP-3) en el cocultivo de células mononucleares de sangre periférica (PBMC) sensibilizadas por secretoma de células madre mesenquimales (MSCS) cocultivadas con células madre de osteosarcoma (OS-SC).

Métodos: Este estudio fue verdaderamente experimental con un diseño de grupo de control solo posterior a la prueba. Este fue un estudio in vitro de PBMSC sensibilizadas por MSCS, ya que luego las muestras se dividieron en 4 grupos de tratamiento, respectivamente: Tratamiento de día cero (P0) Las PBMC se cocultivaron con OS-SC durante 0 horas; Las PBMC del primer tratamiento (P1) se cocultivaron con OS-SC durante 1 hora; Las PBMC del segundo tratamiento (P2) se cocultivaron con OS-SC durante 2 días; Las PBMC del tercer tratamiento (P3) se cocultivaron con OS-SC durante 4 días. El método de examen utilizado en este estudio fue la citometría de flujo y el ensayo inmunoabsorbente ligado a enzimas indirecto (ELISA). Los datos fueron analizados estadísticamente con análisis de varianza (ANOVA) con p≤0.05 considerado como diferencia significativa.

Resultados: Hubo una tendencia a un aumento significativo en la secreción extravesicular en la secreción de IL-2, IL-6, IL-10, CXCL13, TIMP3, en las microvesículas de PBMC cuando se sensibilizan con el secretoma de MSC cocultivado con el entorno de OS-SC. cocultivados después del cuarto día con diferencias significativas entre grupos (p≤0.05).

Conclusiones: Las microvesículas de PBMSC como IL-2, IL-6, IL-10, CXCL13, TIMP3 fueron significativamente sensibilizadas por MSCS y cocultivadas con OS-SC después del cuarto día de in vitro.

Palabras Clave: células madre; enfermedad no transmisible; enfermedad no infecciosa; medicina; osteosarcoma.

Citation Format: Prawiragara FA, Ferdiansyah, Edward M, Utomo DN, Basuki MH, Nugraha AP, Rantam FA (2022) Activation of microvesicle peripheral blood mononuclear cells by mesenchymal stem cells secretome co-cultivated with osteosarcoma stem cell. J Pharm Pharmacogn Res 10(5): 782–790.
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