Avaliação da expressão gênica de citocinas e fatores neurotróficos em células tronco mesenquimais caninas pré-condicionadas com extrato de canabis rico em canabidiol
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Data
2023-05-24
Autores
Perino, Vinicius Skau
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Universidade Estadual Paulista (Unesp)
Resumo
As células-tronco mesenquimais (MSCs) são caracterizadas como um tipo de célula-tronco multipotente conhecidas por possuir a capacidade de diferenciação em diversas linhagens celulares, modulação da secreção de moléculas bioativas (fatores neurotróficos, citocinas, entre outros), atua na reparação e regeneração tecidual, características que permitem sua aplicação em doenças degenerativas, imunomediadas e expressa seu enorme potencial dentro da medicina regenerativa. O uso terapêutico da canabis tem experimentado um crescimento significativo ao longo dos anos, em grande parte devido aos efeitos benéficos dos fitocanabinóides, como o canabidiol (CBD) e o Δ9-tetrahidrocannabinol (THC), dentre outros componentes da planta como os terpenos e flavonóides. Estudos têm demonstrado que o sistema endocanabinoide é responsável por funções biológicas cruciais, como neuroproteção, imunomodulação, modulação da dor e inflamação. O pré-condicionamento das células-tronco mesenquimais derivadas de tecido adiposo canino (cAT-MSCs) utilizando extrato de canabis rico em CBD é uma área emergente de pesquisa que explora os potenciais efeitos dos fitocanabinóides de aprimorar suas propriedades terapêuticas (imunomodulação/ efeitos anti-inflamatórios/ viabilidade e sobrevivência celular/ diferenciação e potencial Regenerativo). O CBD e o THC são os componentes mais estudados e desempenham um importante papel na proliferação, migração, diferenciação e modulação de citocinas e fatores neurotróficos das MSCs in vitro. O objetivo deste estudo in vitro foi avaliar a morfologia e viabilidade celular, assim como a expressão gênica de fatores neurotróficos (BDNF, GDNF e HGF) e citocinas (IL-10, IDO, TNF-α, INF-ɣ e PTGE2) após a estimulação das cAT-MSCs com extrato de canabis rico em CBD. Cinco amostras diferentes de cAT-MSCs, previamente isoladas, caracterizadas imunofenotipicamente (CD44+, CD90+, CD105+, CD34-, CD45-, MHC-II) e diferenciadas (osteogênica, adipogênica e condrogênica), foram pré-condicionadas com concentrações altas (2,25 µM) e baixas (225 nM) do extrato de cannabis rico em CBD por 24 horas. As cAT-MSCs estimuladas com baixa e alta concentração de CBD não apresentaram alterações na morfologia ou na atividade metabólica. O presente trabalho sugere que o perfil de expressão dos fatores neurotróficos (HGF e BDNF) das cAT-MSCs pode ser modulado pelos fitocanabinóides presentes no extrato de canabis rico em CBD, no entanto, o pré-condicionamento das cAT-MSCs com extrato de canabis rico em CBD é uma área de pesquisa relativamente nova, sendo necessário a realização de novos estudos para uma melhor análise do comportamento das cAT-MSCs frente a diferentes extratos, doses e tempo de estimulação in vitro.
Mesenchymal stem cells (MSCs) are characterized as a type of multipotent stem cell known for their ability to differentiate into various cell lineages and modulate the secretion of bioactive molecules (neurotrophic factors, cytokines, among others). They also play a role in tissue repair and regeneration, making them applicable to degenerative and immune-mediated diseases, and showcasing their enormous potential in regenerative medicine. The therapeutic use of cannabis has experienced significant growth over the years, largely due to the beneficial effects of phytocannabinoids such as cannabidiol (CBD) and Δ9-tetrahydrocannabinol (THC), as well as other components of the plant like terpenes and flavonoids. Studies have demonstrated that the endocannabinoid system is responsible for crucial biological functions such as neuroprotection, immunomodulation, pain modulation, and inflammation. Preconditioning canine adipose tissue-derived mesenchymal stem cells (cAT-MSCs) using CBD-rich cannabis extract is an emerging area of research that explores the potential effects of phytocannabinoids in enhancing their therapeutic properties (immunomodulation, anti-inflammatory effects, viability and cell survival, differentiation, and regenerative potential). CBD and THC are the most studied components and play an important role in the proliferation, migration, differentiation, and modulation of cytokines and neurotrophic factors of MSCs in vitro. The objective of this in vitro study was to evaluate the cellular morphology and viability, as well as the gene expression of neurotrophic factors (BDNF, GDNF, and HGF) and cytokines (IL-10, IDO, TNF-α, INF-ɣ, and PTGE2) after stimulating cAT-MSCs with CBD-rich cannabis extract. Five different samples of cAT-MSCs, previously isolated, immunophenotypically characterized (CD44+, CD90+, CD105+, CD34-, CD45-, MHC-II), and differentiated (osteogenic, adipogenic, and chondrogenic), were preconditioned with high (2.25 μM) and low (225 nM) concentrations of CBD-rich cannabis extract for 24 hours. The cAT-MSCs stimulated with low and high concentrations of CBD showed no changes in morphology or metabolic activity. This study suggests that the expression profile of neurotrophic factors (HGF and BDNF) in cAT-MSCs can be modulated by the phytocannabinoids present in CBD-rich cannabis extract. However, preconditioning cAT-MSCs with CBD-rich cannabis extract is a relatively new area of research, and further studies are needed to better analyze the behavior of cAT-MSCs in response to different extracts, doses, and duration of in vitro stimulation.
Mesenchymal stem cells (MSCs) are characterized as a type of multipotent stem cell known for their ability to differentiate into various cell lineages and modulate the secretion of bioactive molecules (neurotrophic factors, cytokines, among others). They also play a role in tissue repair and regeneration, making them applicable to degenerative and immune-mediated diseases, and showcasing their enormous potential in regenerative medicine. The therapeutic use of cannabis has experienced significant growth over the years, largely due to the beneficial effects of phytocannabinoids such as cannabidiol (CBD) and Δ9-tetrahydrocannabinol (THC), as well as other components of the plant like terpenes and flavonoids. Studies have demonstrated that the endocannabinoid system is responsible for crucial biological functions such as neuroprotection, immunomodulation, pain modulation, and inflammation. Preconditioning canine adipose tissue-derived mesenchymal stem cells (cAT-MSCs) using CBD-rich cannabis extract is an emerging area of research that explores the potential effects of phytocannabinoids in enhancing their therapeutic properties (immunomodulation, anti-inflammatory effects, viability and cell survival, differentiation, and regenerative potential). CBD and THC are the most studied components and play an important role in the proliferation, migration, differentiation, and modulation of cytokines and neurotrophic factors of MSCs in vitro. The objective of this in vitro study was to evaluate the cellular morphology and viability, as well as the gene expression of neurotrophic factors (BDNF, GDNF, and HGF) and cytokines (IL-10, IDO, TNF-α, INF-ɣ, and PTGE2) after stimulating cAT-MSCs with CBD-rich cannabis extract. Five different samples of cAT-MSCs, previously isolated, immunophenotypically characterized (CD44+, CD90+, CD105+, CD34-, CD45-, MHC-II), and differentiated (osteogenic, adipogenic, and chondrogenic), were preconditioned with high (2.25 μM) and low (225 nM) concentrations of CBD-rich cannabis extract for 24 hours. The cAT-MSCs stimulated with low and high concentrations of CBD showed no changes in morphology or metabolic activity. This study suggests that the expression profile of neurotrophic factors (HGF and BDNF) in cAT-MSCs can be modulated by the phytocannabinoids present in CBD-rich cannabis extract. However, preconditioning cAT-MSCs with CBD-rich cannabis extract is a relatively new area of research, and further studies are needed to better analyze the behavior of cAT-MSCs in response to different extracts, doses, and duration of in vitro stimulation.
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Palavras-chave
Fitocanabinóides, Sistema endocanabinóide, Imunomodulação, Priming de repetição, Células-tronco mesenquimais, Cannabis, Cães, Canabidiol