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. 2025 Sep 16;75(6):103906. doi: 10.1016/j.identj.2025.103906

Evaluating the Effects of Selective Serotonin Reuptake Inhibitors on Osteogenesis of Periodontal Stem Cells: An In Vitro Study

Yang Zeng a,⁎, Sha Li b, Jun Jiang b,⁎
PMCID: PMC12466211  PMID: 40961628

Abstract

Objectives

Selective serotonin reuptake inhibitors (SSRIs) are commonly prescribed antidepressants, but accumulating evidence suggests they may impair bone metabolism. This study aimed to investigate the effects of SSRIs on the osteogenic differentiation of human periodontal ligament stem cells (PDLSCs) and elucidate the underlying molecular mechanisms, with potential implications for dental regenerative therapies.

Methods

In this in vitro study, human PDLSCs were isolated from extracted premolars and treated with varying concentrations of fluoxetine during osteogenic induction. Osteogenesis was evaluated by ALP and Alizarin Red staining. Transcriptomic and proteomic analyses were conducted to identify differentially expressed genes and proteins, followed by validation using ELISA and Western blotting. Rescue experiments involved IGF-1 and HGF overexpression. A comparative analysis of 5 SSRIs was also performed.

Results

Fluoxetine significantly inhibited PDLSC osteogenesis in a dose-dependent manner without inducing cytotoxicity, with ALP activity reduced by 45.2% at 15 μM (p < .001) and calcium nodule formation decreased by 52.8% (p < .001). Among the SSRIs tested, sertraline and paroxetine showed the strongest inhibitory effects (p < .001), while citalopram and fluvoxamine exhibited minimal effects. Integrated omics analysis revealed downregulation of IGF-1 and HGF, and their overexpression partially rescued the impaired osteogenic differentiation.

Conclusions

This study suggests that SSRIs may impair osteogenic differentiation of PDLSCs by suppressing IGF-1 and HGF signaling pathways. These findings provide mechanistic insight into SSRI-induced bone suppression and support the consideration of antidepressant selection in patients requiring bone or periodontal tissue regeneration.

Key words: Periodontal ligament stem cells (PDLSCs), Selective serotonin reuptake inhibitors (SSRIs), Osteogenic differentiation, IGF-1, HGF, Dental tissue regeneration

Introduction

Serotonin (5-hydroxytryptamine, 5-HT) is a monoamine neurotransmitter that plays a critical role in the nervous system and brain functions. Beyond its regulation of mood and cognition, serotonin is also extensively involved in bone remodeling and periodontal tissue repair.1 Its biological effects on bone and dental tissues are primarily mediated by modulating bone cell activity and molecular signaling networks, thereby influencing stem cell-driven dental repair processes. Specifically, upon tooth injury, platelets are activated and release serotonin, which binds to serotonin receptor 2B (5-HT2BR) on dental pulp stem cells, triggering a series of downstream signaling cascades. Among these pathways, the tissue-nonspecific alkaline phosphatase (TNAP) pathway plays a key role by promoting bone mineralization,2 facilitating cementum formation,3 and further contributing to dental tissue repair.4

SSRIs are a class of antidepressants that selectively block the neuronal reuptake of serotonin, enhancing serotonergic neurotransmission. Common SSRIs include fluoxetine, paroxetine, sertraline, fluvoxamine, and citalopram. Compared with tricyclic antidepressants and monoamine oxidase inhibitors, SSRIs are more widely used in clinical practice due to their higher safety and fewer side effects. Nevertheless, long-term SSRI use may cause various adverse effects, including gastrointestinal discomfort, headache, and drowsiness. Importantly, SSRIs have been reported to induce bone loss, which is believed to involve 2 main mechanisms: first, they may affect the apoptosis of osteoblasts and osteoclasts5; second, SSRIs may enhance central serotonin-dependent sympathetic output, promoting bone resorption.6 Previous studies have reported conflicting evidence regarding SSRI effects on bone metabolism. While some clinical studies suggest increased fracture risk with long-term SSRI use,7,8 the underlying cellular mechanisms, particularly in dental tissues, remain poorly understood. Specifically, limited research exists on SSRI effects on periodontal stem cells, with most studies focusing on general bone metabolism rather than dental-specific regenerative processes.

Building upon these general bone metabolism findings, in the field of dentistry, SSRIs have been implicated in marginal bone loss around dental implants,9 potentially impairing osseointegration.10 The process of osseointegration depends on bone formation and remodeling, in which PDLSCs play a pivotal role. PDLSCs possess multipotent differentiation potential (osteogenic, adipogenic, and chondrogenic lineages) and exhibit high proliferative and self-renewal capacities,11 making them an ideal cell source for periodontal tissue regeneration.12 Therefore, investigating the effects of SSRIs on PDLSCs, particularly their osteogenic differentiation, will help elucidate the potential impact of SSRIs on osseointegration and provide valuable information for clinical medication choices.

In this study, we selected fluoxetine as a representative SSRI to investigate its effect on the osteogenic differentiation of PDLSCs and to elucidate the potential molecular mechanisms involved. Furthermore, we compared the differential effects of 5 commonly used SSRIs on PDLSC osteogenesis to provide preclinical evidence for safer antidepressant selection in dental patients (Figure 1).

Fig. 1.

Fig 1

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Experimental workflow of the study. (A) Osteogenic differentiation of PDLSCs was induced with osteogenic medium and fluoxetine was added at the same time. (B) The effect of fluoxetine on osteogenic differentiation of PDLSCs was evaluated by alkaline phosphatase staining and Alizarin red staining. (C) Proteomic and transcriptomic analysis of fluoxetine-treated PDLSCs. (D) Screening of differentially expressed proteins (DEPs) and differentially expressed genes (DEGs) of PDLSCs under the action of fluoxetine. (E) Analysis of DEPs and DEGs to screen key targets. (F) Overexpression of IGF-1 and HGF in PDLSCs to reverse the osteogenesis inhibition caused by fluoxetine. (G) Comparison of the effects of 5 SSRIs on osteogenic differentiation of PDLSCs to provide preclinical basis for drug selection.

Materials and methods

Isolation and culture of human PDLSCs

This study was approved by the Ethics Committee of the Affiliated Stomatological Hospital of Southwest Medical University (Approval No.: 20210323001). The isolation and culture of PDLSCs were performed as previously described.13 Human premolars extracted for orthodontic purposes were collected with informed consent. Periodontal ligament tissues were scraped from the middle third of the root surface and digested with 0.1% type I collagenase at 37°C for 30 minutes. The tissues were then cultured in Minimum Essential Medium (MEM) supplemented with 10% fetal bovine serum. When the cells reached 80% confluence, they were passaged using trypsin. Flow cytometry was used to characterize PDLSCs, confirming positive expression of CD90, CD105, and STRO-1, and negative expression of CD34 and CD45.14 Cells with a purity >95% were used for subsequent experiments.

Osteogenic induction

PDLSCs were cultured in osteogenic induction medium. Alkaline phosphatase (ALP) staining was performed on day 7 using a BCIP/NBT ALP staining kit (Beyotime), and Alizarin Red S (ARS) staining was performed on day 21 using an ARS staining kit (Solarbio) to assess osteogenic differentiation. The staining results were observed and recorded under a microscope. Semi-quantitative analysis was performed according to the manufacturer's instructions by measuring absorbance values after dye extraction.

Effect of fluoxetine on PDLSC viability and osteogenic differentiation

PDLSCs were cultured in osteogenic induction medium supplemented with fluoxetine at concentrations of 5, 10, or 15 μM (Sigma-Aldrich, F-918). The concentration range was determined based on previous studies5 and pre-experiment results to ensure observable effects without significant cytotoxicity. After 24 hours, cell viability was assessed using Cell Counting Kit-8 (CCK-8, Dojindo, Japan). On day 7, ALP staining and semi-quantitative analysis were performed. On day 21, ARS staining and semi-quantitative analysis were conducted.

Transcriptomic and proteomic analyses

PDLSCs were cultured in osteogenic induction medium with 10 μM fluoxetine for 21 days. Cells were washed with PBS and collected in 500 μL PBS. After centrifugation at 1000 rpm for 20 minutes at 4°C, the supernatant was removed, and the cell pellet was lysed with 1 mL TRIzol reagent. Three biological replicates were set for each group (control and fluoxetine-treated). Proteomic analysis was performed by Lumingbio (Shanghai, China). Total proteins were extracted, quantified, and subjected to SDS-PAGE quality control, trypsin digestion, and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Transcriptomic analysis was also conducted by Lumingbio. Total RNA was extracted, mRNA was enriched, fragmented, and reverse transcribed to cDNA, followed by library construction and RNA-seq. Three biological replicates were included per group.

Bioinformatics analysis

Differentially expressed genes (DEGs) and proteins (DEPs) in PDLSCs treated with fluoxetine were analyzed. Gene Ontology (GO) enrichment analysis was performed using the GO database (https://geneontology.org/). Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis was performed using the KEGG database (https://www.genome.jp/kegg/). Protein-protein interaction (PPI) network analysis was conducted using the STRING database (https://string-db.org/).

Screening of osteogenesis-related genes

Proteomic and transcriptomic results were integrated, and DEPs/DEGs with a fold change >2 and P < .05 were selected. A list of osteogenesis-related genes was obtained from GeneCards (https://www.genecards.org/) and compared with the identified DEPs/DEGs to screen for osteogenesis-related targets affected by fluoxetine.

Overexpression of IGF-1 and HGF

PDLSCs (5 × 10^3 cells) were seeded in 96-well plates containing 100 μL DMEM. When cell confluence reached 40%-60%, PDLSCs were infected with lentiviruses overexpressing IGF-1 and HGF (LV-IGF-1 and LV-HGF, GeneChem, Shanghai, China) at a multiplicity of infection (MOI) of 30 for 16 hours. The culture medium was replaced, and the cells were further cultured for 72 hours. Infection efficiency was evaluated using fluorescence microscopy, and IGF-1 and HGF overexpression was confirmed by ELISA and Western blot. Stable overexpressing clones were selected using 400 μg/mL G418 and 2 μg/mL puromycin.

Comparison of different SSRIs on PDLSC function

Five SSRIs (citalopram, fluvoxamine, fluoxetine, paroxetine, sertraline; Sigma-Aldrich) were evaluated for their effects on PDLSC viability and osteogenic differentiation. Stock solutions were prepared in methanol and diluted to working concentrations of 5, 10, and 15 μM. Cell viability was assessed using CCK-8 assay. Osteogenic differentiation was evaluated using ALP and ARS staining.

Statistical analysis

All experiments were performed with at least 3 independent biological replicates. Data are presented as mean ± SD. Statistical analysis was conducted using GraphPad Prism 9.0 (GraphPad Software). Student's t-test was used for comparisons between 2 groups, and one-way analysis of variance (ANOVA) followed by Tukey's post hoc test was used for multiple group comparisons. Normality was assessed by the Shapiro-Wilk test, and homogeneity of variance was evaluated by Levene's test. P-values less than .05 were considered statistically significant. Significance levels were denoted as follows: *P < .05, **P < .01, ***P < .001.

Results

Effect of fluoxetine on PDLSC viability

PDLSCs were treated with different concentrations of fluoxetine (5, 10, and 15 μM). CCK-8 assay results showed a concentration-dependent decrease in PDLSC viability (Figure 2). However, even at the highest concentration of 15 μM, cell viability remained above 85%, indicating low cytotoxicity within this concentration range.

Fig. 2.

Fig 2

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Effect of fluoxetine on PDLSC viability. PDLSCs were treated with different concentrations of fluoxetine (5, 10, and 15 μM) for 24 hours. Cell viability was assessed using the CCK-8 assay. Data are presented as mean ± SD (n = 3). *P < .05, **P < .01 compared with the control group.

Effect of fluoxetine on osteogenic differentiation of PDLSCs

ALP and ARS staining were performed to assess osteogenic differentiation.15 Under osteogenic induction, PDLSCs showed positive ALP staining on day 7 and ARS staining on day 21. Fluoxetine treatment led to a dose-dependent reduction in ALP (Figure 3A and B) and ARS (Figure 3C and D) staining intensity, suggesting an inhibitory effect on osteogenesis.

Fig. 3.

Fig 3

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Effect of fluoxetine on osteogenic differentiation of PDLSCs. (A) Under the induction of osteogenic medium, the ALP level of PDLSCs increased significantly, while after fluoxetine treatment, the ALP level decreased in a dose-dependent manner. (B) Semi-quantitative analysis of ALP activity. (C) ARS staining was used to evaluate the calcium nodule formation ability of PDLSCs. Under the induction of osteogenic medium, the ARS level of PDLSCs increased significantly, while after fluoxetine treatment, the ARS staining level decreased in a dose-dependent manner. (D) Semi-quantitative analysis of ARS staining. Data are presented as mean ± SD (n = 3). *P < .05, **P < .01, ***P < .001 compared with the control group.

Proteomic and transcriptomic characteristics of fluoxetine-treated PDLSCs

To investigate the potential molecular mechanisms by which fluoxetine inhibits the osteogenic differentiation of PDLSCs, we treated PDLSCs undergoing osteogenic induction with 10 μM fluoxetine and performed proteomic analysis to examine their protein expression profiles. The results revealed that fluoxetine treatment significantly altered the protein expression patterns of PDLSCs. Based on the fold change and statistical P values of the differentially expressed proteins (DEPs), we selected the top-ranked key proteins for further analysis (Figure 4A).

Fig. 4.

Fig 4

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Proteomic and transcriptomic analyses of PDLSCs treated with fluoxetine. (A) Protein samples of PDLSCs were separated by SDS-PAGE electrophoresis, and the differences in protein expression between the control group and the fluoxetine-treated group were compared by proteomic analysis. Based on the protein spectrum results, the most significantly different DEPs were screened for further analysis. (B) The changes in mRNA expression of PDLSCs between the control group and the fluoxetine-treated group were analyzed by RNA-seq. Based on the significance of the expression difference, the key DEGs were screened for subsequent analysis. (C) The protein interaction network (PPI) was constructed using the STRING database. First, the PPI network of all DEPs was constructed, and then the DEPs related to osteogenic differentiation were screened according to gene function to construct a specific PPI network. Finally, the key DEPs were further screened by literature review, and finally an osteogenic-related PPI network with IGF1 and HGF as the core was constructed.

In addition, to comprehensively elucidate the effects of fluoxetine on PDLSCs, we conducted RNA sequencing (RNA-seq) to analyze their mRNA expression profiles. The results similarly demonstrated that fluoxetine treatment significantly affected gene expression in PDLSCs. According to the fold change and statistical P values of the differentially expressed genes (DEGs), we selected the top-ranked key genes for subsequent analysis (Figure 4B). Gene Ontology (GO) enrichment analysis indicated that these affected genes were mainly involved in biological processes such as extracellular matrix formation and cell adhesion. Furthermore, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that these genes were closely associated with various cellular metabolic pathways.

To explore the interactions among the DEPs, we performed protein-protein interaction (PPI) network analysis using the STRING database. First, a comprehensive PPI network was constructed using all identified DEPs. Subsequently, based on functional annotations from the GeneCards database, we screened for osteogenesis-related DEPs and constructed an osteogenesis-related PPI network. Finally, through in-depth analysis, we established an osteogenesis-related PPI network centered on IGF-1 and HGF (Figure 4C).

Fluoxetine inhibits PDLSC osteogenesis by downregulating IGF-1 and HGF

By searching the GeneCards database and reviewing relevant experimental literature (Table 1), we selected IGF-1 and HGF from the DEPs as the core targets for further investigation, as both play important regulatory roles in the process of osteogenic differentiation.

Table 1.

Osteogenesis/osteoclastogenesis-related genes/proteins downregulated by fluoxetine treatment and their roles in dental bone remodeling.

Gene Role in PDLSCs or other dental-derived stem cells
IGF-1 (Insulin-like growth factor 1) Promotes osteogenic differentiation and osteogenesis of PDLSCs16 and stem cells from apical papilla.17,18 Also enhances proliferation and osteogenic differentiation of human dental pulp stem cells (hDPSCs)19 and dental pulp stem cells (DPSCs).20
CCN4 (Cellular communication network factor 4) The role of CCN proteins in musculoskeletal development and physiology has been widely studied.21,22 CCN4 regulates chondrogenic differentiation of human bone marrow stromal cells (hBMSCs).23
IGFBP5 (Insulin-like growth factor binding protein 5) Promotes osteogenic differentiation of PDLSCs24 and DPSCs,25 and enhances periodontal tissue regeneration.26,27
HGF (Hepatocyte growth factor) Promotes PDLSC proliferation28 and periodontal bone regeneration.29
FAM20C (Gene encoding a protein kinase targeting S-X-E/pS motifs) Regulates the differentiation of mesenchymal stem cells into functional odontoblasts and controls dentin mineralization.30, 31, 32, 33
TNFRSF11B (TNF receptor superfamily member 11B) Inhibits osteoclastogenesis34 and promotes chondrocyte-to-osteoblast differentiation.35
WNT5A (Wnt Family Member 5A) Inhibits osteogenic differentiation of PDLSCs.36
SFRP1 (Secreted frizzled-related protein 1) Acts as a Wnt antagonist and an important regulator of mineral homeostasis in periodontal tissues. It is highly expressed in PDLSCs37 and promotes their differentiation.38
TGFB2 (Transforming growth factor beta-2) Inhibits osteogenic differentiation of PDLSCs.39
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To validate this hypothesis, we treated PDLSCs with fluoxetine at concentrations ranging from 0 to 15 μM during osteogenic induction and assessed IGF-1 expression by ELISA and HGF expression by Western blot. The results showed that the expression levels of IGF-1 and HGF gradually decreased with increasing concentrations of fluoxetine (Figure 5).

Fig. 5.

Fig 5

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Fluoxetine inhibits the expression of IGF-1 and HGF in PDLSCs. ELISA and Western blot results showed that the expression levels of IGF-1 and HGF gradually decreased with increasing concentrations of fluoxetine. Data are presented as mean ± SD. *P < .05; **P < .01; ***P < .001.

Overexpression of IGF-1 and HGF reverses fluoxetine-induced inhibition of osteogenic differentiation

To validate the roles of IGF-1 and HGF, PDLSCs were infected with lentiviruses overexpressing IGF-1 and HGF. Results showed that overexpression of IGF-1 and HGF significantly restored ALP activity (Figure 6A and B) and calcium nodule formation (Figure 6C and D) suppressed by fluoxetine treatment.

Fig. 6.

Fig 6

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Overexpression of IGF-1 and HGF reverses fluoxetine-induced inhibition of osteogenic differentiation. (A) ALP staining showed that overexpression of IGF-1 and HGF restored the decreased ALP activity induced by fluoxetine treatment. (B) Semi-quantitative analysis of ALP activity. (C) ARS staining demonstrated that overexpression of IGF-1 and HGF significantly reversed the inhibition of calcium nodule formation caused by fluoxetine. (D) Semi-quantitative analysis of ARS staining. Data are presented as mean ± SD. **P < 0.01; ***P < 0.001.

Effects of different SSRIs on PDLSC osteogenesis

This study compared the effects of 5 commonly used SSRIs (citalopram, fluvoxamine, fluoxetine, paroxetine, and sertraline) on the viability and osteogenic differentiation of PDLSCs.

By assessing the effects of these drugs at different concentrations on PDLSC viability, we found that sertraline exhibited a significant inhibitory effect on cell viability. After treatment with 10 μM sertraline for 24 hours, the viability of PDLSCs decreased to 60% (Figure 7). Based on these findings, a concentration of 10 μM was used for citalopram, fluvoxamine, fluoxetine, and paroxetine in subsequent experiments, while the concentration of sertraline was adjusted to 5 μM due to its stronger cytotoxicity.

Fig. 7.

Fig 7

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Effects of 5 SSRIs on the viability of PDLSCs. Different concentrations of 5 SSRIs were added to the osteogenic induction medium of PDLSCs, and cell viability was assessed after 24 hours. All SSRIs exhibited varying degrees of inhibitory effects on PDLSC viability in a dose-dependent manner. However, fluvoxamine, fluoxetine, paroxetine, and citalopram showed relatively weaker inhibitory effects, whereas sertraline exerted a significant inhibitory effect. Data are presented as mean ± SD.

ALP and ARS staining results demonstrated differential effects of SSRIs on PDLSC osteogenesis. Sertraline and paroxetine exhibited the strongest inhibitory effects, while citalopram showed the weakest (Figure 8A-D). ELISA and Western blot analyses of IGF-1 and HGF expression levels showed consistent trends with osteogenic differentiation results (Figure 8E).

Fig. 8.

Fig 8

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Effects of 5 SSRIs on the osteogenic differentiation of PDLSCs. (A and B) ALP staining and semi-quantitative analysis showed that citalopram had the weakest inhibitory effect on ALP activity of PDLSCs, while sertraline and paroxetine exhibited the most significant inhibitory effects. (C and D) ARS staining and semi-quantitative analysis reflected calcium nodule formation. Citalopram had the least inhibitory effect on calcium nodule formation, whereas sertraline and paroxetine exerted the strongest inhibitory effects. (E) The levels of IGF-1 in PDLSCs treated with different SSRIs were detected by ELISA, and HGF expression was analyzed by Western blot. The results showed that citalopram had the weakest effect on the expression of these proteins, while sertraline and paroxetine exhibited the most significant inhibitory effects, which were consistent with the ALP and ARS staining results. Data are presented as mean ± SD. Statistical significance: *P < .05; **P < .01; ***P < .001.

Discussion

SSRIs are commonly prescribed antidepressants due to their safety and efficacy.40 However, recent studies have reported that long-term SSRI use may adversely affect bone metabolism, increasing the risk of osteoporosis and fractures.41 In the field of dentistry, SSRIs are suspected of interfering with alveolar bone remodeling and impairing osseointegration of dental implants,9,10 although the underlying mechanisms remain unclear.

Recent years have witnessed increasing attention to pharmacological regulation of dental stem cells. Wang et al.42 and Yang et al.43 respectively reported the promoting effects of valproic acid (VPA) and ornidazole on dental stem cell differentiation, which presents an interesting contrast to our findings of SSRI inhibitory effects, suggesting that different pharmaceuticals exert distinctly different impacts on dental regeneration and further emphasizing the importance of rational drug selection in dental patients.

In this study, we systematically evaluated the effects of fluoxetine on PDLSC osteogenic differentiation. Our results demonstrated that fluoxetine inhibited PDLSC osteogenic differentiation in a dose-dependent manner without significant cytotoxicity. The fluoxetine concentrations used in this study (5-15 μM) are higher than typical therapeutic plasma concentrations (approximately 0.35-1.45 μM, equivalent to 120-500 ng/mL),44,45 but tissue concentrations can be significantly higher due to extensive tissue binding and distribution, with brain-to-plasma ratios reaching 2.6:1 and other tissues showing even higher distribution coefficients.46 Proteomic and transcriptomic analyses revealed that fluoxetine treatment significantly downregulated the expression of 2 key osteogenesis-related factors, IGF-1 and HGF. Further experiments confirmed that overexpression of IGF-1 and HGF effectively reversed fluoxetine-induced inhibition of osteogenic differentiation, suggesting that these growth factors play pivotal roles in SSRI-mediated suppression of PDLSC osteogenesis.

IGF-1 and HGF have been reported to regulate the proliferation and osteogenic differentiation of various dental stem cells.16, 17, 18, 19, 20,28,29 IGF-1 promotes PDLSC osteogenic differentiation through activation of ERK and JNK MAPK pathways,16 while HGF enhances cell proliferation and osteogenic gene expression, contributing to alveolar bone regeneration.28,29 Our findings indicate that fluoxetine may impair PDLSC osteogenic microenvironment by downregulating IGF-1 and HGF, thereby inhibiting osteogenesis. While our findings provide mechanistic insights into SSRI effects on periodontal stem cells, direct clinical extrapolation requires caution. The in vitro environment lacks the complex interactions present in vivo, including immune responses, vascular supply, and systemic factors that may modulate SSRI effects on bone metabolism.

Although this study elucidated the inhibitory effects of fluoxetine on IGF-1 and HGF expression, the upstream regulatory mechanisms remain unclear. Previous studies have suggested that SSRIs suppress 5-HT signaling and interfere with cAMP/PKA and ERK/MAPK pathways,5,6 which are closely related to the transcriptional regulation of IGF-1 and HGF.16,28 In addition, SSRIs may affect PI3K/AKT signaling, inhibiting osteoblast survival and function.47 We speculate that SSRI-mediated inhibition of PDLSC osteogenesis may involve coordinated regulation of 5-HT/cAMP/PKA, MAPK, and PI3K/AKT pathways, leading to downregulation of IGF-1 and HGF expression. These pathways require further investigation. The temporal effects of SSRI therapy on bone metabolism remain complex. While our study examined acute exposure effects, clinical bone changes typically develop over months to years of treatment. Short-term SSRI therapy may have different effects than long-term exposure, potentially involving adaptive cellular responses and changes in bone turnover markers over time.

Furthermore, we compared the effects of 5 commonly used SSRIs on PDLSC osteogenesis. Our results showed that citalopram and fluvoxamine exerted relatively weaker inhibitory effects, while sertraline and paroxetine exhibited the strongest suppression. These differences may be attributed to variations in 5-HT transporter affinity and downstream signaling effects. These findings suggest that clinicians should consider the potential adverse effects of SSRIs on bone metabolism when selecting antidepressants for patients requiring dental procedures.

Several limitations should be acknowledged. First, this study was conducted entirely in vitro, lacking the complex bone microenvironment present in vivo. Second, we examined only acute effects of SSRI exposure, while clinical effects may require prolonged treatment. Third, the upstream signaling mechanisms regulating IGF-1 and HGF suppression require further investigation. Fourth, individual patient factors such as age, gender, and concurrent medications may influence SSRI effects on bone metabolism. Future studies should include animal models to validate these findings and long-term exposure studies to assess reversibility of effects.

Collectively, this study revealed that SSRIs, particularly fluoxetine, inhibit PDLSC osteogenic differentiation by downregulating IGF-1 and HGF expression. We also compared the differential effects of various SSRIs on PDLSC osteogenesis. These findings provide experimental evidence for rational selection of SSRIs in clinical settings, especially for patients undergoing alveolar bone remodeling treatments. Further studies are warranted to elucidate upstream signaling networks and verify biological effects in animal models and clinical patients. These findings may provide insights into the safe clinical use of SSRIs in dental patients and inform future strategies for periodontal tissue regeneration.

Conclusion

This in vitro study suggests that SSRIs may inhibit osteogenic differentiation of PDLSCs through downregulation of IGF-1 and HGF expression. While these findings provide preliminary evidence for potential SSRI effects on periodontal regeneration, further validation through animal studies and clinical research is essential before making treatment recommendations. For patients requiring both antidepressant therapy and dental procedures, individualized assessment considering the risk-benefit ratio may be warranted.

Conflict of interest

None disclosed.

Acknowledgments

Author contributions

Jun Jiang and Yang Zeng conceived the research and drafted the manuscript. Sha Li performed the experiments. Jun Jiang analyzed the data and revised the manuscript. All authors reviewed and approved the final manuscript.

Funding

This study was supported by the National Natural Science Foundation of China (82070288), the Sichuan Province Science & Technology Program (2022YFS0627), and Luzhou City and Southwest Medical University Collaborative Research Program (2024LZXNYDJ019).

Ethical approval

This study was approved by the Ethics Committee of the Affiliated Stomatological Hospital of Southwest Medical University (Approval No.: 20210323001). Written informed consent was obtained from all participants.

Contributor Information

Yang Zeng, Email: zengyang826@swmu.edu.cn.

Jun Jiang, Email: jiangjun@swmu.edu.cn.

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