Research ArticleExperimental Studies
Open Access

Cannabidiol Enhances SIRT1 and Autophagy for the Maintenance of Human Mesenchymal Stem Cells

PHONGSAKORN CHUEAPHROMSRI, PHONGSAKORN KUNHORM, AREECHUN SOTTHIBUNDHU, NIPHA CHAICHAROENAUDOMRUNG and PARINYA NOISA
In Vivo January 2026, 40 (1) 222-234; DOI: https://doi.org/10.21873/invivo.14186

Abstract

Background/Aim: Mesenchymal stem cells (MSCs) are used to treat various degenerative diseases. However, their therapeutic potential is limited by cellular aging during in vitro cultivation. This study aimed to explore whether cannabidiol (CBD) can delay MSC aging by enhancing the expression of Sirtuin 1 (SIRT1) and autophagy, two key anti-aging regulators.

Materials and Methods: CBD, the most important non-psychotomimetic phytocannabinoid derived from the Cannabis sativa plant, was used to up-regulate SIRT1 and autophagy in order to maintain MSC stemness. MSCs were treated with CBD and assessed for cell viability, doubling time, key gene/protein expression, relative senescence-associated β-galactosidase (SA-β-gal) assay, relative telomere length, and telomerase expression.

Results: CBD significantly increased the expression of SIRT1 and autophagy-related markers in MSCs. Furthermore, CBD preserved MSC stemness by promoting the deacetylation of SRY-box transcription factor 2 (SOX2) through SIRT1, and delayed cellular senescence by enhancing autophagy, reducing SA-β-gal activity, maintaining proliferation capacity, and supporting telomere function.

Conclusion: CBD promotes MSC stemness and delays cellular senescence, potentially through the activation of SIRT1 and autophagy. These findings suggest that CBD may serve as a promising agent for preserving MSC function in regenerative medicine.

Keywords:

Introduction

Modern medical advancements, including cell therapy and regenerative medicine, have garnered significant attention as innovative approaches for treating severe diseases (1). Mesenchymal stem cells (MSCs) have been utilized to address a wide range of degenerative conditions, such as cardiovascular disease, neurodegenerative disorders, bone and cartilage diseases, cancer, liver and kidney disorders, graft-versus-host disease, multiple sclerosis, Crohn’s disease, type 1 diabetes, systemic lupus erythematosus, and rheumatoid arthritis (2). Classified as adult stem cells, MSCs are commonly derived from bone marrow, adipose tissue, umbilical cord tissue, and umbilical cord blood (3). They possess the ability to develop and differentiate into various cell types, including adipocytes, osteoblasts, chondrocytes, endothelial cells, and cardiomyocytes (4). The defining stemness characteristics of MSCs – self-renewal and multipotency (5) – are crucial for the development of specialized tissues and organs with distinct functions (6). However, these stemness properties decline with aging, posing a major challenge to the therapeutic application of MSCs in degenerative diseases (7). When cultured in vitro over extended periods, MSCs progressively lose their stemness and become unsuitable for clinical use (8). Moreover, replicative senescence leads to long-term changes in phenotype, reduced differentiation potential, and alterations in global gene expression and microRNA profiles – all of which are key considerations for MSC rejuvenation strategies (9). Therefore, identifying effective methods to preserve MSC self-renewal and multipotency is critical for their clinical application.

SIRT1 is a nicotinamide adenine dinucleotide (NAD+)-dependent lysine deacetylase that participates in numerous biological processes, including gene silencing, DNA repair, metabolic regulation, cell cycle control, apoptosis, inflammation, autophagy, and cellular senescence (10). By deacetylating key proteins, SIRT1 directly or indirectly modulates age-related signaling pathways such as FoxO1, NF-κB, AMPK, mTOR, p53, and PGC-1, thereby delaying cellular senescence (11). A growing body of evidence indicates that enhanced SIRT1 activity may decelerate aging and associated diseases in mammals by regulating DNA damage and metabolic decline (12). Furthermore, SIRT1 has been implicated in maintaining stemness through the regulation of pluripotency transcription factors such as NANOG and OCT4 in both embryonic stem cells and MSCs (13). Conversely, SIRT1 suppression in MSCs has been shown to reduce cellular proliferation and promote senescence (14). Thus, strategies aimed at upregulating SIRT1 may be beneficial for maintaining MSC properties.

Autophagy is a fundamental degradative process that enables adaptive responses to metabolic stress, such as nutrient deprivation. It functions to eliminate potentially harmful cellular components, including protein aggregates and dysfunctional organelles, thereby maintaining cellular homeostasis (15). Enhanced autophagy is associated with anti-aging effects, whereas impaired autophagy has been linked to accelerated aging (16).

Cannabidiol (CBD) is the major non-psychotomimetic phytocannabinoid derived from the Cannabis sativa plant (17). Numerous studies have demonstrated its broad pharmacological effects, including antidepressant, anti-inflammatory, antiemetic, neuroprotective, analgesic, antibacterial, anticonvulsant, anxiolytic, antipsychotic, antitumor, and immunomodulatory activities (18). Recently, CBD has been shown to extend lifespan and improve health span in various models, including Caenorhabditis elegans and Danio rerio (19). Moreover, CBD induces autophagy in SH-SY5Y cells, protecting them against mitochondrial dysfunction through the upregulation of SIRT1, which inhibits the NF-κB and NOTCH pathways (20). Therefore, CBD is considered a promising bioactive compound due to its ability to promote SIRT1 and potentially preserve MSC properties.

In this study, we employed cytotoxicity assays and key gene/protein expression analyses to optimize the CBD concentration required for SIRT1 upregulation in MSCs. We further examined key molecular markers to investigate how CBD influences autophagy activation in MSCs. Additionally, we developed CBD-supplemented culture media aimed at preserving MSC self-renewal and multipotency by integrating an optimal CBD concentration into standard media. The anti-aging effects of the formulated culture media were evaluated using doubling time, relative senescence-associated β-galactosidase (SA-β-gal) activity, relative telomere length, and telomerase activity.

Materials and Methods

MSCs and cell culture. MSCs were obtained from I WELLNESS Co., Ltd. (Nakhon Ratchasima, Thailand) in accordance with ethical guidelines. The cells were cultured in α-minimum essential medium (α-MEM) (Cytiva, Marlborough, MA, USA) supplemented with 10% (v/v) fetal bovine serum (FBS) (Gibco, Carlsbad, CA, USA), 100 U/ml penicillin (Gibco), 1 mM L-glutamine (Gibco), and 1 mM non-essential amino acids (Gibco). The MSCs were maintained at 37°C in a humidified incubator with 5% CO2.

CBD preparation. CBD (≥95% pure) was obtained from the Biorefinery Pilot Plant, Suranaree University of Technology (Nakhon Ratchasima, Thailand) as a purified extract. The CBD stock solution was prepared by dissolving it in 99.8% absolute ethanol (Sigma-Aldrich, St. Louis, MO, USA) to a final concentration of 1,000 μg/ml. The solution was filtered through a 0.2 μm syringe filter (Millex®, Millipore, Burlington, MA, USA) prior to incorporation into the standard culture medium for experimental use.

Cell viability assay. MSCs were seeded in 96-well plates (Corning Inc., Corning, NY, USA) at a density of 2,000 cells per well and allowed to adhere for 24 h. The cells were then treated with CBD at concentrations of 0.04, 0.08, 0.16, 0.31, 0.63, and 1.25 μg/ml. Untreated MSCs served as the control group. Cell viability was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT; Sigma-Aldrich) assay according to the procedure described by Meerloo et al. (21). Absorbance at 570 nm was measured using a microplate reader (BMG Labtech, Ortenberg, Germany). Cell viability in the control group was considered 100%, and the half-maximal inhibitory concentration (IC50) of CBD was calculated using the online calculator provided by AAT Bioquest Inc. (22).

Effects of CBD on messenger ribonucleic acid (mRNA) expression in MSCs using real-time-polymerase chain reaction (RT-PCR). MSCs were seeded in 6-well plates (Corning) at a density of 30,000 cells per well and incubated for 24 h. The cells were then treated with CBD at concentrations of 0.04 and 0.08 μg/ml for 24 h, while untreated MSCs served as the control group. After treatment, cells were harvested using 0.025% trypsin-EDTA (Sigma-Aldrich). Total RNA was extracted using the NucleoSpin® RNA kit (MACHEREY-NAGEL, Dueren, Germany) according to the manufacturer’s instructions. cDNA was synthesized using ReverTra Ace™ qPCR RT Master Mix with gDNA Remover (TOYOBO, Osaka, Japan). Quantitative real-time PCR was performed using qPCRBIO SyGreen Mix (PCR Biosystems, Wayne, PA, USA) on a QuantStudio™ 5 Real-Time PCR System (Thermo Fisher Scientific, Waltham, MA, USA), following the procedure described by Olveira et al. (23). GAPDH was used as the housekeeping gene. Primers for SIRT1, SOX2, OCT4, NANOG, p53, mTOR, NF-κB, LC3, Beclin 1, ATG5, and ATG12 were used to amplify target sequences as listed in Table I.

View this table:
Table I.

PCR primer sets used for gene expression analysis.

Immunofluorescence staining (IF). MSCs were seeded on glass slides in 24-well plates (Corning) at a density of 15,000 cells per well and incubated for 24 h. The cells were then treated with 0.08 μg/ml CBD for 24 h, while untreated MSCs served as the control group. After treatment, cells were fixed with Fixing Solution I (4% paraformaldehyde and 400 mM sucrose in PBS; Sigma-Aldrich) for 30 min at 37°C. Fixing Solution II (Fixing Solution I containing 0.5% Triton X-100; Sigma-Aldrich) was then applied for 15 min at room temperature. Slides were washed with PBS and blocked with 0.5% bovine serum albumin (BSA; Sigma-Aldrich) in PBS for 1 h at room temperature. Following three washes with PBS, slides were incubated overnight at 4°C with primary antibodies: anti-phospho-SIRT1 (Affinity Biosciences, Cincinnati, OH, USA) or anti-LC3-I/II (Merck, Darmstadt, Germany) at a 1:500 dilution. After five washes with cold PBS, slides were incubated with Alexa Fluor 488-conjugated goat anti-rabbit IgG (H+L; 1:500; Sigma-Aldrich) and then mounted with DAPI (Invitrogen, Waltham, MA, USA) to visualize nuclei. Fluorescence images were captured using a ZOE™ Fluorescent Cell Imager (Bio-Rad, Singapore).

Relative SA-β-gal assay. MSCs were seeded in 96-well plates (Corning) at a density of 2,000 cells per well and cultured in medium containing 0.08 μg/ml CBD or without CBD as the control. After four days, cells were washed twice with PBS and fixed with freshly prepared 3.7% formaldehyde in PBS (Sigma-Aldrich) for 5 min at room temperature. Following two additional PBS washes, cells were incubated with 100 μL of X-gal staining solution prepared according to Itahana et al. (24). Cells were then incubated at 37°C (without CO2) for 15 h. Relative SA-β-gal activity was quantified by measuring absorbance at 420 nm using a microplate reader (BMG Labtech). MSC passages 3, 5, 7, and 9 from continuous cultures were used to assess relative SA-β-gal activity.

Doubling time. MSCs were seeded in T75 flasks (Corning) at a density of 500,000 cells per flask. Cells were cultured in medium containing 0.08 μg/ml CBD or without CBD as the control. After four days, cells were harvested using 0.025% trypsin-EDTA (Sigma-Aldrich) and counted using a Bright-Line™ Hemocytometer (Sigma-Aldrich). Doubling time was calculated using the online tool provided by Omni Calculator (25). MSC passages 3, 5, 7, and 9 from continuous cultures were used to assess doubling time.

Relative telomere length assessment using RT-PCR. MSC genomic DNA was extracted using a Mouse Direct PCR Kit (Selleck Chemicals LLC, Houston, TX, USA). Quantitative analyses were performed using the qPCRBIO SyGreen Mix (PCR Biosystems) and the QuantStudio™ 5 Real-Time PCR System (Thermo Fisher Scientific). The ribosomal protein gene 36B4, which serves as a single-copy reference gene, and telomere-specific (TEL) primers were used to assess the telomere-to-single copy gene (T/S) ratio according to the procedure described by Vasilishina et al. (26) (Table I).

Telomerase activity measurement by RT-PCR. Total RNA from MSCs was extracted using the NucleoSpin® RNA kit (MACHEREY-NAGEL) according to the manufacturer’s instructions. cDNA was synthesized using ReverTra Ace™ qPCR RT Master Mix with gDNA Remover (TOYOBO). Quantitative real-time PCR was performed using qPCRBIO SyGreen Mix (PCR Biosystems) on a QuantStudio™ 5 Real-Time PCR System (Thermo Fisher Scientific), following the procedure described by Olveira et al. (22). GAPDH was used as the housekeeping gene, and hTERT primers were employed to evaluate telomerase activity according to Liu et al. (27) (Table I).

Statistical analysis. All statistical analyses were performed using IBM SPSS Statistics software (IBM Corp., Armonk, NY, USA). IF images were analyzed using ImageJ software (National Institutes of Health, Bethesda, MD, USA). Data are presented as mean±standard deviation (SD). Differences between groups were evaluated using the independent-sample t-test. Statistical significance was defined as *p<0.05 (*p≤0.05; **p≤0.01).

Results

The cytotoxicity of CBD on MSCs. To evaluate the cytotoxic effects of CBD on MSCs, cells were treated with increasing concentrations of CBD (0.04, 0.08, 0.16, 0.31, 0.63, and 1.25 μg/ml), while untreated MSCs served as the control group (Figure 1A). CBD at concentrations ≥0.16 μg/ml significantly decreased MSC viability, whereas lower concentrations (0.04 and 0.08 μg/ml) promoted a significant increase in cell viability compared with the control. The IC50 of CBD in MSCs was calculated to be 0.37 μg/ml. Moreover, MSCs treated with CBD exhibited no noticeable alterations in morphology, maintaining comparable cell size and flat, fibroblast-like morphology relative to the control group (Figure 1B).

Figure 1.
Figure 1.

The effect of cannabidiol (CBD) on mesenchymal stem cells (MSCs). (A) The MTT assay was used to assess the cytotoxicity of CBD on MSCs. AAT Bioquest website was utilized to determine the IC50. (B) The morphology of MSCs treated with and without CBD. The images were digitally captured at 10X magnification (scale bar: 30 μm). (C) The mRNA expression levels of SIRT1, SOX2, OCT4, NANOG, p53, mTOR, and NF-κB were examined using RT-PCR in MSCs treated with CBD for 24 h. Data is presented as mean±SD. Independent-sample t-test was used to analyze group differences (n=3). Statistical significance is indicated by *p<0.05 (*p≤0.05; **p≤0.01).

The effect of CBD on the expression of SIRT1 and aging-related mRNAs in MSCs. The mRNA expression levels of SIRT1, SOX2, OCT4, NANOG, p53, mTOR, and NF-κB in MSCs were quantified by RT-PCR (Figure 1C). CBD treatment significantly upregulated genes associated with MSC stemness and multipotency, including SIRT1, SOX2, OCT4, and NANOG, with the most pronounced increase observed at 0.08 μg/ml. Conversely, CBD markedly downregulated the expression of aging- and stress-related genes, namely p53, mTOR, and NF-κB, with maximal suppression also occurring at 0.08 μg/ml, compared with untreated controls.

The effect of CBD on phosphorylated SIRT1 (p-SIRT1) protein levels in MSCs. The protein expression of p-SIRT1 in MSCs was assessed using IF (Figure 2). Treatment with 0.08 μg/ml CBD resulted in a significant increase in p-SIRT1 protein levels compared with the untreated control group.

Figure 2.
Figure 2.

The effect of cannabidiol (CBD) on the levels of p-SIRT1 protein expression in MSCs. p-SIRT1 protein levels were measured using immunofluorescent staining, and staining intensity was assessed using ImageJ. DAPI was used to counterstain the cell nuclei. A fluorescence microscope was used to digitally capture the images (scale bar: 30 μm). Data is presented as mean±SD. Independent-sample t-test was used to analyze group differences (n=3). Statistical significance is indicated by *p<0.05 (*p≤0.05; **p≤0.01).

The effect of CBD on autophagy in MSCs. The expression levels of LC3, Beclin1, ATG5, and ATG12 mRNAs in MSCs were assessed using RT-PCR (Figure 3A), and LC3 protein levels were analyzed by IF (Figure 3B). CBD treatment significantly upregulated the mRNA expression of LC3, Beclin1, ATG5, and ATG12 compared with the control group, with the most pronounced effect observed at 0.08 μg/ml. Furthermore, 0.08 μg/ml CBD markedly increased LC3 protein levels in MSCs relative to the control group.

Figure 3.
Figure 3.

The effect of cannabidiol (CBD) on autophagy in mesenchymal stem cells (MSCs). (A) The mRNA expression levels of LC3, Becline1, ATG5, and ATG12 were examined using RT-PCR in MSCs treated with CBD for 24 h. (B) The protein levels of LC3 were determined using immunofluorescent staining. ImageJ was used to assess staining intensity. DAPI was used to stain the cell nuclei. The images were captured digitally using a fluorescence microscope (scale bar: 30 m). Data is presented as mean±SD. Independent-sample t-test was used to analyze group differences (n=3). Statistical significance is indicated by *p<0.05 (*p≤0.05; **p≤0.01).

The anti-aging effects of CBD-supplemented medium on MSCs. To investigate the anti-aging effects of CBD, MSCs were cultured in media supplemented with 0.08 μg/ml CBD from passages 3 to 9, while the control group was maintained in CBD-free medium. Relative SA-β-gal activity in MSCs was assessed using X-gal staining, in which enzymatic reaction with β-galactosidase results in a color change from yellow to blue (Figure 4A). Optical density at 420 nm progressively decreased from passages 3 to 9 in both groups (Figure 4B), reflecting relative SA-β-gal activity. The CBD-treated group exhibited significantly lower relative SA-β-gal activity than the control group at passages 5 and 7, whereas no significant differences were observed at passages 3 and 9. Additionally, doubling time progressively increased from passages 3 to 9 in both groups (Figure 4C). However, CBD-treated MSCs demonstrated a significantly shorter doubling time compared to the control group at passages 5 and 7, with no significant differences detected at passages 3 and 9. Furthermore, MSCs in the CBD group preserved relative telomere length more effectively than the control group at passage 7 (Figure 5A). Consistently, the CBD group showed a significant increase in hTERT expression compared with the control group at passage 7 (Figure 5B).

Figure 4.
Figure 4.

The anti-aging properties of cultural media containing cannabidiol (CBD). (A) The mesenchymal stem cell (MSC) passage 7 reacted with an X-gal staining solution for 15 h after being grown in a medium containing CBD or without CBD. (B) Relative SA-β-gal activity of MSCs from passages 3 to 9 cultured in medium with or without CBD. Relative SA-β-gal activity was determined by measuring optical density (OD) at 420 nm using a microplate reader. (C) The doubling time of MSCs from passage 3 to 9 after being grown in a medium containing CBD or without CBD. Data is presented as mean±SD. Independent-sample t-test was used to analyze group differences (n=3). Statistical significance is indicated by *p<0.05 (*p≤0.05; **p≤0.01).

Figure 5.
Figure 5.

Impact of cannabidiol (CBD) on mesenchymal stem cells’ (MSCs) relative telomere length and telomerase activity. (A) The telomere-to-single copy gene (T/S) ratio was examined using RT-PCR in MSC passage 7 that were cultured in a medium with CBD or without CBD. (B) The expression of hTERT was examined using RT-PCR in MSC passage 7 which was cultured in a medium with CBD or without CBD. Data is presented as mean±SD. Independent-sample t-test was used to analyze group differences (n=3). Statistical significance is indicated by *p<0.05 (*p≤0.05; **p≤0.01).

Discussion

Our study demonstrates that CBD at an appropriate concentration enhances the proliferation of MSCs, consistent with previous findings showing that CBD promotes MSC migration, proliferation, and osteogenic differentiation (28). We further observed that CBD upregulates both gene and protein levels of SIRT1 and autophagy-related markers in MSCs. This effect can be mechanistically linked to AMPK activation (Figure 6). Cannabinoids, a group of bioactive compounds that includes CBD, activate AMPK by inducing cellular stress, including ER stress, which increases the cellular AMP/ATP and ADP/ATP ratios (29). The γ subunit of AMPK senses this energy imbalance, leading to phosphorylation of Thr172 on the α subunit by LKB1, thereby fully activating AMPK (29). In some contexts, CBD may also activate Ca2+/PKA-dependent pathways to further stimulate AMPK signaling (30). Activated AMPK promotes SIRT1 activity by increasing cellular NAD+ levels (31), while SIRT1 can reciprocally enhance AMPK activation, reflecting a positive feedback loop between the two regulators (32, 33).

Figure 6.
Figure 6.

Proposed mechanism by which cannabidiol (CBD) promotes mesenchymal stem cell (MSC) stemness and delays cellular aging. This schematic summarizes the signaling pathways through which CBD enhances SIRT1 activity in MSCs, leading to the maintenance of stemness properties and the delay of cellular senescence.

Our results demonstrated that CBD promotes autophagy in MSCs by upregulating key autophagy-related genes and proteins, including LC3, Beclin 1, ATG5, and ATG12. The induction of autophagy appears to be mediated by SIRT1, which negatively regulates mTOR signaling through its interaction with the TSC1/2 complex (34). In addition, SIRT1 facilitates autophagosome formation by deacetylating essential autophagy regulators, thereby enhancing the autophagic process and contributing to cellular homeostasis (34).

CBD was observed to upregulate the stemness-associated transcription factors SOX2, OCT4, and NANOG in MSCs, thereby contributing to the maintenance of stem cell properties. This finding is consistent with previous reports demonstrating that SOX2, OCT4, and NANOG are essential for preserving the self-renewal and multipotency of MSCs (35-38). Yoon et al. further proposed that SIRT1 serves as a key regulator of MSC stemness by modulating the expression of these transcription factors through deacetylation, which prevents their nuclear export and subsequent ubiquitination and degradation in the cytoplasm (13). In addition, SOX2 within the nucleus can regulate OCT4 and NANOG expression, thereby reinforcing the transcriptional network required to sustain MSC self-renewal and multipotency (39).

SIRT1 has been identified as a key target for promoting healthy longevity, as it integrates multiple signaling and transcriptional pathways, including p53, mTOR, and NF-κB, all of which are critically involved in the regulation of aging (11). In our study, we observed that CBD upregulates SIRT1 expression while concurrently downregulating p53 expression in MSCs, suggesting a protective role against DNA damage and stress-induced cellular senescence (40). Furthermore, our results revealed that CBD reduces mTOR expression in MSCs, potentially through SIRT1-mediated deacetylation of tuberous sclerosis complex 2 (TSC2), thereby promoting autophagy and enhancing cellular longevity (41, 42). Additionally, we found that CBD suppresses NF-κB expression in MSCs, possibly via SIRT1-mediated deacetylation of p65, contributing to the attenuation of inflammatory responses and age-related processes (43).

Finally, the anti-aging effects of CBD-supplemented medium were evaluated by assessing relative SA-β-gal activity, population doubling time, relative telomere length, and telomerase activity. SA-β-gal activity is a widely recognized biomarker of cellular senescence (44), and our findings indicated that CBD delays the onset of senescence in MSCs. Additionally, CBD was found to preserve the proliferation rate of MSCs, as evidenced by an improved doubling time. The ability of CBD to sustain MSC proliferation suggests that it supports the maintenance of self-renewal and multipotency, given that aging is associated with a progressive decline in proliferative capacity (45). Moreover, CBD increased hTERT gene expression, thereby preventing telomere shortening in MSCs. Progressive telomere attrition leads to senescence, apoptosis, or oncogenic transformation in somatic cells, all of which negatively impact health and lifespan (46). Although CBD can preserve MSC characteristics and delay cellular aging, prolonged culture ultimately results in senescence. Therefore, CBD serves as a potential agent for preventing or postponing MSC aging and cellular senescence.

Conclusion

This study demonstrates that an optimal concentration of CBD enhances MSC proliferation and promotes SIRT1 activation, thereby inducing autophagy and maintaining stemness through the regulation of SOX2. Moreover, CBD was found to delay cellular senescence and preserve replicative potential in MSCs. Collectively, these findings highlight CBD as a promising modulatory agent for improving MSC longevity and therapeutic quality, with potential implications for regenerative and anti-aging applications.

Acknowledgements

This research received funding support from (i) Suranaree University of Technology (SUT) and (ii) Thailand Science Research and Innovation (TSRI), and (iii) the National Science, Research and Innovation Fund (NSRF), (NRIIS number 179316).

Footnotes

  • Authors’ Contributions

    P.C. and P.N. designed the study. MSCs were supplied by P.K., N.C., and P.N., who also provided essential resources and materials. All experiments were carried out by P.C. Data analysis was performed by P.C., P.K., A.S., and N.C. The interpretation of experimental findings was conducted by P.C., P.K., and N.C. P.C. prepared all figures and drafted the manuscript. Manuscript revisions and critical editing were contributed by P.K., A.S., and N.C. The final revision and approval of the manuscript were completed by P.N.

  • Conflicts of Interest

    There are no conflicts of interest related to this study.

  • Artificial Intelligence (AI) Disclosure

    During the preparation of this manuscript, a large language model (QuillBot) was used solely for language editing and stylistic improvements in select paragraphs. No sections involving the generation, analysis, or interpretation of research data were produced by generative AI. All scientific content was created and verified by the authors. Furthermore, no figures or visual data were generated or modified using generative AI or machine learning–based image enhancement tools.

  • Received September 11, 2025.
  • Revision received October 15, 2025.
  • Accepted October 16, 2025.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY-NC-ND) 4.0 international license (https://creativecommons.org/licenses/by-nc-nd/4.0).

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Cannabidiol Enhances SIRT1 and Autophagy for the Maintenance of Human Mesenchymal Stem Cells
PHONGSAKORN CHUEAPHROMSRI, PHONGSAKORN KUNHORM, AREECHUN SOTTHIBUNDHU, NIPHA CHAICHAROENAUDOMRUNG, PARINYA NOISA
In Vivo Jan 2026, 40 (1) 222-234; DOI: 10.21873/invivo.14186
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