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. 2026 Jul 3;17:333. doi: 10.1186/s13287-026-05135-w

SERINC3 promotes osteogenic differentiation of BMSCs via IL-32/AMPK-mediated autophagy and mitochondrial energy metabolism

Zhichao Zheng 1,#, Tianru Xu 1,#, Janak L Pathak 1,#, Shaofen Xu 1, Jiarui Lu 1, Shuiqing Yu 1, Zhihe Fu 1, Wei Xie 1, Haohui Zhu 1, Richard T Jaspers 1,2, Huade Zheng 3,4,5,✉, Lihong Wu 1,✉, Jiang Li 1,✉
PMCID: PMC13613861  PMID: 42400028

Abstract

Background

SERINC3, a member of the serine incorporator protein family, is known for its roles in viral resistance and tumorigenesis, however, its function in osteogenesis remains unexplored.

Methods

Lentivirus infection, alkaline Phosphatase/Alizarin Red S Staining, and RT-qPCR were used to evaluate the osteogenic differentiation of mesenchymal stem cells mediated by SERINC3. MicroCT, H&E, and Masson staining were performed to investigate the bone formation and bone defect repair via Serinc3 knockout (KO) mice and nude mice. RNA sequencing, Co-IP, Western blotting, and Seahorse energy metabolism analysis were performed to elucidate the regulatory mechanism of SERINC3.

Results

Here, we identify SERINC3 as a critical regulator of osteogenic differentiation of bone marrow-derived stem cells (BMSCs) and bone regeneration. SERINC3 expression was significantly upregulated during osteogenic differentiation of BMSCs and stem cells from human exfoliated deciduous teeth (SHED). Functional assays revealed that SERINC3 overexpression enhanced osteogenic differentiation, proliferation, and migration of MSCs, while Serinc3-KO impaired these processes and led to osteopenia in mice. In a calvarial defect model, Serinc3-KO mice exhibited 42% less bone volume (BV/TV) and 35% lower bone mineral density (BMD), whereas SERINC3-overexpressing BMSCs significantly improved bone repair. Mechanistically, RNA sequencing and pathway analysis revealed that SERINC3 interacts with IL32 to activate the AMPK-ULK1-autophagy axis, thereby promoting osteogenesis. Additionally, SERINC3 enhanced mitochondrial energy metabolism by upregulating tricarboxylic acid cycle enzymes (ACO1, DLAT, SDHA) and increasing oxygen consumption rates. Rescue experiments confirmed that AMPK inhibition or autophagy blockade abolished SERINC3-mediated osteogenic effects, whereas mitochondrial electron transport chain activators restored osteogenesis in SERINC3-knockdown cells.

Conclusions

In summary, this study identifies SERINC3 as a novel regulator of bone formation that orchestrates osteogenesis through IL32-AMPK-autophagy signaling axis and mitochondrial metabolism. These findings highlight SERINC3 as a potential therapeutic target for enhancing bone regeneration and treating skeletal defects.

Supplementary Information

The online version contains supplementary material available at https://doi.org/10.1186/s13287-026-05135-w.

Keywords: SERINC3, Osteogenic differentiation, Bone regeneration, BMSCs, AMPK pathway, Autophagy, Energy metabolism

Introduction

Large-area bone defects resulting from trauma, cancer therapy, or other pathologies severely impair patients’ quality of life [1, 2]. When a segmental bone defect exceeds 2 to 2.5 times the bone diameter, the natural healing capacity becomes significantly compromised [3], with 5–10% of fractures failing to heal properly [4]. Current clinical treatments, such as autologous and allogeneic bone transplantation, face major limitations, including donor scarcity, immune rejection, and infection risks [5, 6]. Tissue-engineered bone, which integrates mesenchymal stem cells (MSCs), scaffolds, and cytokines, offers a promising alternative for bone regeneration [7]. Among MSCs, bone marrow-derived stem cells (BMSCs) are the most extensively studied due to their multipotent differentiation potential, making them ideal candidates for bone repair [8]. While existing research has explored various regulatory mechanisms, including immune modulation, metabolic pathways, and growth factors, in BMSC osteogenic differentiation [9–11]. Further identification of key regulatory genes remains crucial for developing novel therapeutic targets for bone regeneration.

The Serine Incorporator (SERINC) protein family, which comprises five members (SERINC1-5), plays a critical role in cellular membrane biology [12]. These proteins share a conserved helix-loop-helix structure across eukaryotes and function primarily to incorporate serine into membranes, facilitating the biosynthesis of phosphatidylserine (PS) and sphingolipids (SP) [13, 14]. These lipids are key regulators of osteogenesis: PS promotes MSC osteogenic differentiation via ERK signaling [15], while SP homeostasis is essential for bone mineralization and metabolism [14, 16]. Notably, PS-based biomaterials (e.g., hydrogels and bioglass) are widely used to enhance bone regeneration [17–19]. Beyond bone biology, SERINCs possess antiviral properties against HIV, MLV, and coronaviruses [20–23] and are implicated in cancer and epilepsy [12, 24]. Despite these diverse roles, the function of this family in osteogenic differentiation remains unexplored, representing a significant knowledge gap.

This study aimed to investigate the regulatory role and underlying mechanisms of SERINC3 in BMSC osteogenic differentiation and bone regeneration. Through gain- and loss-of-function experiments in vitro and using SERINC3-knockout (KO) mice, we demonstrate that SERINC3 expression is upregulated during osteogenesis and critically enhances the differentiation capacity of BMSC. Mechanistically, we identify that SERINC3 interacts with IL32 to activate the AMPK pathway, thereby promoting autophagy and mitochondrial energy metabolism to drive osteogenic differentiation. Furthermore, SERINC3-modified BMSCs significantly improved the healing of calvarial defects. Our findings establish SERINC3 as a novel regulator of bone formation and highlight its potential as a therapeutic target for bone regeneration strategies.

Methods

The work has been reported in line with the ARRIVE guidelines 2.0.

Cell isolation and culture

BMSCs were purchased from OriCell and cultured in medium (OriCell, HUXMA-90011, China). Human calvarial osteoblasts (hOBs) were obtained from Sciencell and maintained in osteoblast medium (Sciencell, 4601, China). Stem cells from human exfoliated deciduous teeth (SHED) were isolated from the pulp tissue of deciduous teeth and identified as shown in Figure S1. The study involving deciduous teeth was approved by the Ethics Committee of the School and Hospital of Stomatology, Guangzhou Medical University (Approval No.: LCYJ2021017). Written informed consent was obtained from the parents of all participants. SHEDs were cultured in α-MEM medium supplemented with 10% FBS and 1% P/S.

RT-qPCR analysis

Cells were lysed with Trizol (Accurate Biology, AG21102, China) and extracted with isopropanol as previously reported [25]. Total RNA (500 ng) was transcribed with reverse transcriptase (Accurate Biology, cat#AG11701, China). Quantitative PCR was performed using SYBR® Green Premix Pro Taq HS qPCR Kit (Accurate Biology, cat#AG11701, China) on an AriaMx Real-time quantitative PCR machine (Agilent, USA). The PCR reaction conditions were 95℃ for 30 s, followed by 40 cycles at 95℃ for 5 s and 60℃ for 30 s. Fold change relative to the control group was measured using the 2−∆∆Ct method. The primers used are listed in Table S1.

Alizarin red staining (ARS)

BMSCs (25,000 cells/well) and SHED (25,000 cells/well) were seeded at a 48-well plate with osteogenic medium (50 µg/mL vitamin C (Solarbio, IA0530, China), 0.01 µM dexamethasone (Solarbio, cat#D8040, China), and 10 mM β-glycerophosphate (Solarbio, G9422)) and incubated in a 5% CO2 atmosphere at 37℃ for 14 days. The cells were fixed with 4% paraformaldehyde and stained with 1% alizarin red for 10 min. Mineralized nodules were observed under a microscope. For quantitative analysis, the alizarin red-stained mineralized matrix was dissolved in 200 µL of 10% cetylpyridinium chloride monohydrate for 1 h. The optical density of the supernatant (100 µL) was measured at 562 nm using a microplate reader (Biotech).

Alkaline phosphatase (ALP) staining

BMSCs and SHED (2.5 × 104 cells/well) were seeded in 48-well culture plates and cultured in an osteogenic medium for 7 days. While BMSCs were cultured in osteogenic differentiation medium (OriCell, HUXMX-90021, China). The cells were then fixed with 4% paraformaldehyde and stained with an ALP staining kit (Beyotime, C3206, China). Staining was visualized under a stereomicroscope using LAS EZ software (Leica, Germany). ALP staining intensity was further analyzed using ImageJ 1.5.1j8 (NIH, USA).

Plasmid construction and lentivirus preparation

The lentivirus plasmids, including the control plasmid, pReceiver-Lv242-SERINC3 plasmid, were purchased from Genecopoeia. PLKO.1-IL32 shRNA plasmid, pCDNA3.1(+)-3×Flag-SERINC3 and pcDNA3.1(+)-3×HA-IL32 were constructed by Generay.

For lentiviral transduction, HEK293T cells were co-transfected with expression plasmids (control, pReceiver-Lv242-SERINC3, SERINC3 shRNA, PLKO.1-IL27 shRNA) and the packaging plasmids pMD2.VSVG, pMDLg/pRRE, and pRSV-REV using EZ trans transfection reagent (Life-iLab, AC04L092). Fresh lentiviral supernatant was collected after 2 days and used for infection. BMSCs were expanded to 60% confluence prior to lentiviral infection. Cells were changed to fresh medium after 10 h. Subsequent experiments were performed 3 days later. The shRNA sequences were as follows: SERINC3 shRNA: 5’-CCACTTGTTCTTACAAATCGT-3’; IL27 shRNA: 5’-GCTCTTCATGTCCTCTTTCCA-3’.

Cell viability

Cell viability was analyzed using PrestoBlue cell viability reagent (Thermo Fisher, A13261, USA). Control, SERINC3-overexpression (OE), SERINC3-knockdown (KD) BMSCs, and SHED (4 × 103 cells/well) were seeded into 96-well culture plates. After 48 h, the cell viability detection medium was added for 2 h. The optical density (OD) value was measured by a microplate reader at 570 nm with a reference wavelength of 600 nm.

Transwell assay

Control, SERINC3-OE, SERINC3- KD BMSCs (3 × 104 cells) and SHEDs (3 × 104 cells) in 200 µL α-MEM medium were seeded in the upper well of an 8 μm pore transwell. And 600 µL complete medium was added to the lower well. After 24 h, cells were fixed with 4% paraformaldehyde and stained with 1% crystal violet (LEAGENE, DZ0053, China) for 10 min.

Scratch assay

Control, SERINC3-OE, SERINC3-KD BMSCs (1.5 × 105 cells), and SHEDs (1.5 × 105 cells) were cultured in a 6-well plate. After confluence, cells were scratched and further cultured. Pictures were taken at 0 h, 6 h, 12 h, and 24 h.

Mice

Serinc3 global knockout mice (Serinc3-KO) and control C57BL/6J wild-type mice were purchased from Gempharmtech (China). The construction and identification of Serinc3 transgenic mice are shown in Figure S3. The blinded evaluation was used for mouse assignments and analysis. All animal experiments were performed at the animal facility of Guangdong Huawei Testing Co., Ltd., a licensed contract research organization. Ethical approval was obtained from the Animal Ethics Committee of Guangdong Huawei Testing Co., Ltd., as the experiments were conducted on their premises.

Anesthesia and euthanasia

Before establishing the calvarial defect model, mice were anesthetized with 1% pentobarbital sodium at a dose of 150 µL per 20 g body weight, administered intraperitoneally. Prior to bone sample collection, mice were first deeply anesthetized with isoflurane inhalation, followed by cervical dislocation to ensure humane euthanasia.

The isolation of primary BMSCs and osteogenic induction

Euthanized transgenic mice and wild-type male mice (5–6 weeks of age) were immersed in 75% ethanol for 5 min. The femurs and tibiae were then harvested. Primary BMSCs were isolated and expanded as described previously [26]. In brief, bone marrow was flushed out from the tibia and femur and cut into small pieces. Cells were collected by centrifugation, plated into culture flasks, and allowed to adhere for 24 h. Nonadherent cells were removed by washing, and the culture was maintained in DMEM supplemented with 10% FBS and 1% penicillin/streptomycin. Cells were cultured in a 5% CO2 incubator, maintaining a humid atmosphere. Upon reaching 80% confluence, cells were trypsinized and passaged.

Bone phenotype analysis

Bone phenotype analysis was conducted via Micro-CT. Mice were anesthetized with isoflurane (RWD Life Science Co., China), followed by cervical dislocation. The femur with a distal growth plate was collected and fixed in 10% buffered formalin. Micro-CT scanning using Bruker Sky1172 Skyscan (Kontich, Belgium) as previously reported [27]. Relative bone formation parameters, including bone mineral density (BMD), bone volume/total volume (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), and trabecular separation (Tb.Sp), were analyzed.

LPS induced inflammatory bone loss

As our previous report [28], LPS was injected for the evaluation of the inflammatory bone destruction in Serinc3-KO mice and control mice for 2 weeks. Femurs were collected and analyzed by micro-CT.

Western blot analysis

Total protein (20 µg) was added to a 10% SDS-polyacrylamide gel. The protein was transferred to PVDF membranes (Millipore, USA) after electrophoresis and blocked for 1 h with 3% bovine serum albumin. Then PVDF membranes were incubated with the primary antibodies against ALP (Abclonal, Cat#A0514, 1:5000), RUNX2 (CST, Cat#12556, 1:5000), SERINC3 (SAB, Cat#33870, 1:5000), IL32 (Zenbio, Cat#R389356, 1:5000), pi-AMPK (CST, Cat#2535, 1:5000), AMPK (Zenbio, Cat#R380431, 1;5000), pi-ULK1 (CST, Cat#5869, 1:5000), ULK1 (Zenbio, Cat#R381887, 1:5000), p62 (CST, Cat#5114, 1:5000), BECLIN1 (CST, Cat#3738, 1:5000), LC3 (Zenbio, Cat#R381554, 1:5000), and GAPDH (Proteintech, Cat#10494-1-AP, 1:10,000, China) overnight at 4℃. The membranes were further incubated with horseradish peroxidase-conjugated secondary antibody for 1 h and reacted with ECL (Millipore, USA). Finally, photographs were taken using the Tanon-5200 system (Tanon, China).

Mice calvarial bone defect healing

As previously reported, a 3 mm diameter was created on one side of the sagittal suture in 8-week Serinc3-KO mice and wild-type mice (8 mice/group, including 4 male mice and 4 female mice, 1 defect/mouse) [29]. A total of 200 ng BMP2 (10 µL) was applied to a collagen membrane (ZH-BIO, China), which was then inserted into the defect. Calvarial bones were collected and analyzed by Micro-CT after 1 month.

BMSCs-based nude mice calvarial bone defect healing

BMSCs were infected with either a scramble or a SERINC3-OE lentivirus and then cultured in osteogenic medium for 7 days. Cell sheets derived from BMSCs were cultured in osteogenic medium supplemented with 20 µg/mL vitamin C for 7 days. BMSCs (105 cells/sample) were seeded onto a collagen membrane (ZH-BIO, China). Five-week-old male nude mice (n = 6) were purchased from Gempharmtech for calvarial bone defect repair. Following anesthesia, a 3 mm diameter bone defect was created and filled with BMSC cell sheets along with the collagen membrane. After 2 months, calvarial bones were collected and analyzed using Micro-CT.

RNA-Seq

RNA-Seq was performed on BMSCs from the control and BMSCsSERINC3-KD groups using the Illumina Novaseq Xplus PE150 platform (Oebiotech, China). The resulting Fastq data were aligned to the GRCh38.p13 genome using htseq-count (https://htseq.readthedocs.io/en/master/htseqcount.html). Differential gene expression analysis was conducted using DESeq2 (https://bioconductor.org/packages/release/bioc/html/DESeq2.html). SRplot (http://www.bioinformatics.com.cn/en) was used to analyze differentially expressed genes (DEGs) and to perform pathway enrichment analysis. A statistically significant cut-off point was set at P < 0.05.

IL32 structure and binding site prediction

AlphaFold (https://alphafold.com/) was used to predict the structures of the SERINC3 and IL32 proteins. Additionally, ClusPro (https://cluspro.org/help.php) was utilized to predict the direct binding between SERINC3 and IL32.

Co-immunoprecipitation (Co-IP)

Flag-SERINC3, HA-IL32, and control plasmids were purchased from Genentech (China). Flag-SERINC3, HA-IL32, and control plasmids (10 µg) were transfected into HEK293T cells with EZ trans transfection reagent. After 3 days of incubation, 107 cells were collected and lysed for 30 min. After centrifuging at 12,000 g for 10 min, the supernatant was collected. Then, 25 µL balanced anti-Flag Nanobody Magarose Beads (AlpaLifeBio, KTSM1338, China) or balanced anti-HA Nanobody Magarose Beads (ABMagic, MA103) were added into the supernatant and rotated at 4℃ for 3 h. The beads were washed 5 times with PBST and eluted with 1×SDS loading buffer at 95℃ for 10 min. The protein was detected with a Flag (Proteintech, cat#20543-1-AP, RRID: AB_11232216, 1:10,000) and HA (Abclonal, cat#AE105, RRID: ARC59578, 1:10000) antibody.

Confirmation of the role of IL32, glycolysis, and oxidative phosphorylation in the SERINC3-mediated osteogenesis

SERINC3-OE BMSCs were treated with IL32-KD lentivirus, compound C (TargetMol, T1977, 200 nM), bafilomycin A1 (MCE, HY-100558, 1 µM), and rotenone (50 nM, MCE, HY-B1756) to assess the effects of IL32 inhibition, AMPK signaling, autophagy, and oxidative phosphorylation on osteogenesis driven by SERINC3 overexpression. Conversely, SERINC3-KD BMSCs were treated with IL32 (R&D Systems, 4690-IL/CF, 100 ng/mL), lactic acid (MCE, HY-B2227, 10 mM), and CoQ10 (MCE, HY-N0111, 30 µM) to evaluate the rescued function of IL32, glycolysis, and oxidative phosphorylation on the impaired osteogenesis resulting from SERINC3 knockdown.

GFP-LC3 puncta detection

Control and SERINC3-OE BMSCs were infected with GFP-LC3 lentivirus. Then the GFP-LC3 puncta were analyzed using an In Cell Analyzer 2500HS (GE, Germany) and quantified in ImageJ.

Autophagosomes detected by transmission electron microscope (TEM)

Control and SERINC3-OE BMSCs were scratched and centrifuged at 3000 rpm for 10 min. The deposits were incubated with 2.5% glutaraldehyde for 30 min at room temperature. The sample treatment and TEM scanning were further performed by Shiyanjia Co., Ltd. (China).

Mito tracker staining

Control and SERINC3-OE BMSCs were stained with Mitotracker Red Regent (Beyotime, C1035, 1:20000) for 15 min. After PBS washing 3 times, the pictures were taken using a Cell Analyzer 2500HS (GE, Germany). The mitotracker staining intensity was calculated with ImageJ.

Analysis of oxidative phosphorylation

Control, SERINC3-OE, and SERINC3-KD BMSCs (1.3 × 104 cells) were inoculated into an XF24 culture microplate and cultured. Concurrently, the probe plate was hydrated, and the cells were washed with analytical medium (pH 7.4). The culture microplate was then returned to a CO2-free incubator for 1 h. For the mitochondrial stress test, 56 µL of 10× oligomycin was added to well A of the probe plate and 62 µL to well B. Additionally, 69 µL of 10× rotenone/antimycin A was added to well C after adding 10× carbonyl cyanide-4-trifluoromethoxyphenylhydrazone (FCCP). Samples were then analyzed with the Seahorse XFe24 (Agilent, USA).

Statistical analysis

Data are expressed as mean ± SD. Independent nonparametric t-tests were used to compare the two groups. The comparison of mean values across multiple groups was performed using one-way analysis of variance (ANOVA), with each group compared to every other group. Statistical significance was set at p < 0.05, and a difference was considered significant.

Results

SERINC3 upregulates during osteogenic differentiation of precursor cells

To investigate the expression pattern of SERINC family members in osteogenic lineage cells, we first compared their expression in BMSCs and hOBs. Notably, SERINC1 and SERINC3 were significantly upregulated in hOBs compared to BMSCs (Fig. 1A). We then analyzed the expression of SERINC family members during osteogenic differentiation of BMSCs and SHED. As expected, successful osteogenic differentiation was confirmed by increased ALP activity, calcium mineralization, and elevated expression of osteogenic markers ALP, RUNX2, and OCN (Fig. 1B and G). Strikingly, among the SERINC family proteins, SERINC3 was consistently upregulated during osteogenic differentiation in both BMSCs and SHED (Fig. 1H, I), suggesting a potential role of SERINC3 in the osteogenic differentiation of precursor cells.

Fig. 1.

Fig. 1

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SERINC3 expression was increased during the osteogenic differentiation of mesenchymal stem cells. A Expression pattern of SERINC3 family proteins in BMSCs and hOBs. ALP staining at day 7 (B) and ARS staining at day 14 (C) of human BMSCs’ culture. D ALP, RUNX2 and OCN expression in BMSCs at day 7. E ALP staining at day 7 of SHED culture. F ARS staining at day 14 of SHED culture. G ALP, RUNX2 and OCN expression in SHED at day 7. H, I SERINC1-5 expression in BMSCs and SHED during osteogenic differentiation at day 7. Significant difference compared to the control group, *p < 0.05, **p < 0.01, and ***p < 0.001. PM proliferation medium, OM osteogenic medium, BMSCs bone marrow mesenchymal stem cells, SHED stem cells from human exfoliated deciduous teeth

SERINC3 enhances osteogenic differentiation, viability, and migration of MSCs

To assess the functional role of SERINC3 in MSCs, we overexpressed or knocked down SERINC3 in BMSCs and SHED. SERINC3-OE significantly increased ALP production and matrix mineralization (Fig. 2A. B; Figure S2A, B). Consistently, osteogenic markers ALP, RUNX2, and OCN were upregulated in SERINC3-OE cells (Fig. 2C; Figure S2C). Conversely, SERINC3-KD reduced mineralization (Fig. 2D, E; Figure S2D, E) and downregulated osteogenic gene expression (Fig. 2F; Figure S2F). Beyond differentiation, SERINC3 also affected cell viability, with overexpression promoting proliferation and knockdown suppressing it (Fig. 2G, H; Figure S2G, H). We further demonstrated that apoptosis inhibitor Z-DEVD-FMK did not influence the SERINC3 regulated osteogenic differentiation (Figure S3). Given the importance of MSC migration in bone regeneration [30], we evaluated this phenotype and found that SERINC3-OE enhanced MSCs migration in transwell and scratch assays, whereas SERINC3-KD impaired it (Fig. 2I L; Figure S2I–L). Together, these results demonstrate that SERINC3 positively regulates osteogenic differentiation, viability, and migration of MSCs, suggesting its potential role in MSCs’ function during bone regeneration.

Fig. 2.

Fig. 2

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Overexpression of SERINC3 promoted osteogenic differentiation, cell viability, and cell migration of BMSCs. A Expression pattern of SERINC3 in SERINC3-OE BMSCs. B The staining and quantification of ALP at day 7 and ARS staining at day 14 (n = 3). C The osteogenic markers ALP, RUNX2 and OCN expression levels in SERINC3-OE BMSCs at day 7 (n = 3). D Expression pattern of SERINC3 in SERINC3-KD BMSCs. E The staining and quantification of ARS staining at day 14 (n = 3). F Expression level of osteogenic markers ALP, RUNX2 and OCN in SERINC3-KD BMSCs at day 7 (n = 3). G The cell viability of SERINC3-OE BMSCs (G) and SERINC3-KD BMSCs (H) (n = 6). The migration ability of SERINC3-OE (I) and SERINC3-KD BMSCs (J) by transwell assay (n = 3). The cell migration detection of SERINC3-OE (K) and SERINC3-KD BMSCs (L) by scratch assay. Data are presented as mean ± SD. Significant difference compared to the control group, *p < 0.05, **p < 0.01, and ***p < 0.001. OE overexpression, KD knockdown

Serinc3 deficiency causes osteopenia through impaired osteogenic differentiation

As demonstrated by our analysis of International Mouse Phenotyping Consortium data, which revealed significantly reduced bone mineral density and content in Serinc3-mutated mice (Fig. 3A), we confirmed these findings in our global Serinc3-KO model (Fig. 3A, B). In this model, where micro-CT and histomorphometry analyses revealed osteopenic phenotypes including 28% reduced BV/TV, 32% decreased Tb.N, and 41% increased Tb.Sp, while trabecular thickness and cortical parameters remained normal (Fig. 3B and F). At the cellular level, Serinc3-KO BMSCs exhibited impaired osteogenic potential, with a 65% reduction in ALP activity, a 72% decrease in mineralization, and downregulated expression of osteogenic markers ALP and RUNX2 (Fig. 3G, H). Furthermore, Serinc3-KO aggravated the trabecular bone loss in the LPS-induced inflammatory mouse model (Figure S5). Collectively, these findings establish SERINC3 as a critical regulator of bone homeostasis, maintaining trabecular architecture and promoting MSC osteogenic differentiation.

Fig. 3.

Fig. 3

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Serinc3-KO mice showed a low bone mass phenotype. A IMPC database of BMD and bone mineral content. B H&E staining. C Representative micro-CT images for trabecular bone. D BV/TV, BMD, Tb.N, Tb.Th, and Tb.Sp of trabecular bone (n = 10, 5 male and 5 female mice). E Representative micro-CT images for cortical bone. F BMD and BV/TV of cortical bone (n = 10, 5 male and 5 female mice). G The staining and quantification of ALP and ARS staining of mouse primary BMSCs (n = 3). H Western blot analysis of ALP and RUNX2 (n = 3). Significant difference compared to wild-type mice, *p < 0.05 and **p < 0.01

Serinc3-KO mice show impaired bone regeneration in an orthotopic model

Serinc3 deficiency significantly impairs bone regeneration in a calvarial defect model, as evidenced by multiple quantitative and qualitative assessments. Micro-CT analysis revealed a 42% reduction in new BV/TV and a 35% decrease in BMD in Serinc3-KO mice compared to WT controls at 8 weeks post-injury (Fig. 4A and C). Histomorphometric evaluation showed a 2.8-fold reduction in osteocalcin-positive areas (Fig. 4F) and 61% reduction in collagen deposition (Fig. 4E) in knockout mice, with H&E staining confirming markedly impaired bone matrix formation (Fig. 4D). These coordinated deficits in mineralization, osteoid production, and collagen organization indicate that SERINC3 is essential for proper bone defect healing by regulating both the inorganic and organic bone matrix components.

Fig. 4.

Fig. 4

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SERINC3-KO mice showed compromised healing of bone defects. A Representative micro-CT images. B Local micro-CT images of defects. C BV/TV, BMD, Tb.N, Tb.Th, and Tb.Sp analysis (n = 4). D H&E staining. E Masson staining. F OCN staining. Data are presented as mean ± SD. Significant difference compared to wild-type mice, *p < 0.05

SERINC3-OE BMSC cell sheets showed enhanced bone regeneration potential

We further assessed the bone-regenerative potential of SERINC3. BMSC cell sheets modified by SERINC3 were implanted into a calvarial defect. SERINC3-OE sheets showed increased ALP and RUNX2 levels compared to the control group (Fig. 5A and C). Consistently, SERINC3-OE BMSC sheets exhibited increased bone formation in the calvarial defects (Fig. 5D and E). Furthermore, the trabecular bone parameters BV/TV and Tb.Th were significantly increased, whereas Tb.Sp showed a non-significant decrease (Fig. 5F). H&E and Masson staining revealed increased new bone and collagen deposition in the SERINC3-OE group (Fig. 5G and H). Collectively, these findings demonstrate that SERINC3-OE BMSCs possess superior bone regenerative potential.

Fig. 5.

Fig. 5

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SERINC3-OE BMSCs promoted bone defect healing in nude mice. A BMSCs’ cell sheets. B, C Western blot analysis of ALP and RUNX2 expression in BMSCs sheets (n = 3). D, E Representative micro-CT images. F BV/TV, BMD, Tb.N, Tb.Th, and Tb.Sp analysis (n = 5). G H&E staining. H Masson staining. Data are presented as mean ± SD. Significant difference compared to the control group, *p < 0.05 and **p < 0.01

SERINC3 promotes bone formation by combining with IL-32

The regulatory mechanism of SERINC3 in osteogenic differentiation was further investigated. RNA-Seq revealed that 307 genes were upregulated and 1037 genes were downregulated in SERINC3-KD BMSCs compared to the control group (Fig. 6A). Pathway enrichment analysis indicated that SERINC3-KD predominantly affected cytokine and cell receptor binding pathways (Fig. 6B). And the expression of IL32 in the cytokine-cell receptor binding pathway was significantly reduced (Fig. 6C). Among these, the IL32 expression was significantly reduced (Fig. 6C). We further confirmed that SERINC3-OE increased IL32 gene and protein expression, whereas SERINC3-KD yielded the opposite effect (Fig. 6D and E). Additional experiments showed that SERINC3-KD inhibited the expression of IL32γ isoform, but not IL32α or IL32β (Figure S6). Rescue assay demonstrated that the IL32γ recombinant protein could reverse the inhibitory effect on osteogenesis induced by SERINC3-KD in BMSCs (Fig. 6F and H). The protein structures of SERINC3 and IL32 were predicted using AlphaFold, and their binding was further predicted with ClusPro (Fig. 6I). Co-IP assay confirmed direct interaction between SERINC3 and IL32 (Fig. 6J). Further rescue assay demonstrated that IL-32 knockdown attenuated the osteogenesis-promoting effect of SERINC3 (Fig. 6K and M). Collectively, these findings suggest that SERINC3 may synergize with IL32 to promote osteogenic differentiation in BMSCs.

Fig. 6.

Fig. 6

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SERINC3 forms a functional complex with IL-32 to promote osteogenesis. A Different gene expression. B KEGG enrichment pathway analysis. C Heatmap of differential gene expression in the cytokine-cytokine receptor interaction pathway. D Relative IL-32 gene expression in SERINC3-OE and SERINC3-KD group (n = 3). E Western blot analysis of IL-32 and SERINC3 in SERINC3-OE and SERINC3-KD group (n = 3). F, G ALP staining at day 7 and ARS staining at day 14, and the quantification (n = 3). H Relative ALP and RUNX2 expression (n = 3). I ClusPro prediction of the interaction between SERINC3 and IL-32. J Co-IP analysis of the binding between SERINC3 and IL-32. K, L ALP staining at day 7 and ARS staining at day 14, and the quantification (n = 3). M Relative ALP and RUNX2 expression (n = 3). Significant difference compared to the control group, *p < 0.05, **p < 0.01, and ***p < 0.001. OE overexpression, KD knockdown

SERINC3 promotes BMSC osteogenic differentiation by regulating the AMPK-autophagy pathway

AMPK is an essential regulator of cell viability, the autophagy pathway, and mitochondrial energy metabolism [31–33]. IL32 may ameliorate liver damage in high-fat diet-fed mice by activating AMPK [34]. Furthermore, the AMPK-ULK1-autophagy axis has been demonstrated to regulate osteogenic differentiation in mouse BMSCs [35]. We found that SERINC3-OE increased the levels of phosphorylated AMPK and ULK1 (Fig. 7A, B). Expression of autophagy-related markers LCII/LCI and BECLIN1 was increased, while p62 was decreased (Fig. 7A, B). These data indicated that SERINC3-OE activated the AMPK-ULK1-autophagy pathway. Conversely, SERINC3-KD reduced activation of this pathway (Fig. 7A, B). Rescue assay indicated that AMPK inhibitor Compound C suppressed SERINC3-mediated osteogenesis in BMSCs, as evidenced by ALP staining, calcium deposits formation, and ALP/RUNX2 gene expression (Fig. 7C and E). The AMPK knockdown by lentivirus showed similar results (Figure S7A–C). Furthermore, SERINC3-OE increased the GFP-LC3 expression levels in BMSCs (Fig. 7F), and transmission electron microscopy revealed an increase of autophagosomes in SERINC3-OE BMSCs (Fig. 7F). Additionally, bafilomycin A1, an inhibitor of the autophagy pathway, impaired the SERINC3-mediated pro-osteogenesis effect (Fig. 7G and I). These results suggest that SERINC3 exerts an anabolic effect in osteogenic differentiation of BMSCs by modulating the AMPK-ULK1-autophagy pathway.

Fig. 7.

Fig. 7

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SERINC3 promotes osteogenic differentiation of BMSCs via modulating the AMPK-ULK1-autophagy pathway. A Western blot analysis in SERINC3-OE and SERINC3-KD BMSCs. B The protein expression quantification (n = 3). C, D ALP staining at day 7 and ARS staining at day 14, and the quantification (n = 3). E Relative ALP and RUNX2 expression (n = 3). F GFP-LC3 puncta detection of SERINC3-OE BMSCs (n = 35) and transmission electron microscope analysis of autophagosome in BMSCs. G, H ALP staining at day 7 and ARS staining at day 14 and the quantification (n = 3). I Relative ALP and RUNX2 expression (n = 3). Significant difference compared to the control group, *p < 0.05, **p < 0.01, and ***p < 0.001. OE overexpression

SERINC3 promotes BMSCs osteogenic differentiation by affecting mitochondrial energy metabolism

AMPK activation has been shown to increase the mitochondrial energy metabolism level in MSCs, which has been reported to promote osteogenic differentiation [36].GSEA analysis revealed the enrichment of the tricarboxylic acid cycle in SERINC3-KD BMSCs (Fig. 8A). Consistently, bioinformatic analysis and RT-qPCR analysis showed that SERINC3-KD significantly reduced the expression of TCA-related genes, including CS, ACO1, ACO2, DLAT, DLD, SDHA, and MDH1 (Fig. 8B and D). In contrast, the expression of ACO1, DLAT, and SDHA genes was significantly increased in the SERINC3-OE group (Fig. 8D). Measurement of oxygen consumption rate further revealed that SERINC3-OE increased basal respiration and ATP production (Fig. 8E, F), while SERINC3-KD exhibited the opposite effect (Fig. 8E, F). Mitochondria serve as the core organelle for energy metabolism. Mitochondrial staining intensity was markedly increased in SERINC3-OE BMSCs (Fig. 8G, H). To investigate functional relevance, the mitochondrial electron transport chain in BMSCs was activated or inhibited using CoQ10 or rotenone. CoQ10 treatment improved impaired osteogenic differentiation in BMSCs caused by SERINC3 knockdown (Fig. 8I), whereas rotenone attenuated SERINC3-OE-induced osteogenic differentiation in BMSCs (Fig. 8J). These results suggest that SERINC3 promotes osteogenic differentiation through mitochondrial metabolism. A schematic diagram summarizing the mechanism by which SERINC3 regulates osteogenic differentiation in precursor cells is presented in Fig. 8K.

Fig. 8.

Fig. 8

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SERINC3 regulates mitochondrial energy metabolism to promote osteogenic differentiation of BMSCs. A GSEA analysis. B TCA cycle. C Differential gene expression involved in TCA. D RT-qPCR analysis of TCA-related genes in SERINC3-OE and SERINC3-KD BMSCs. E, F The oxygen consumption rate (OCR) analysis of SERINC3-OE and SERINC3-KD BMSCs. G, H MitoTracker mitochondrial staining and quantification. H ALP staining at day 7, ARS staining at day 14, and the quantification (n = 3). I ALP staining at day 7 and ARS staining at day 14, and the quantification (n = 3). J ALP staining at day 7 and ARS staining at day 14, and the quantification (n = 3). K Graphical summary of the regulatory mechanism of SERINC3 on osteogenic differentiation of BMSCs. SERINC3-upregulated IL32 binds SERINC3 in an autocrine manner, activating the AMPK-mediated autophagy pathway and mitochondrial energy metabolism to promote osteogenic differentiation of precursor cells. Significant difference compared to the control group, *p < 0.05, **p < 0.01, and ***p < 0.001. OE overexpression

Discussion

Our study identifies SERINC3 as a novel regulator of BMSC osteogenesis and bone regeneration, operating through two distinct yet interconnected mechanisms: [1] formation of a functional complex with IL32γ and [2] activation of AMPK-dependent autophagy alongside mitochondrial metabolic reprogramming. These findings substantially expand the biological repertoire of SERINC3 beyond its previously characterized roles in viral restriction [20, 37, 38], positioning it as a new molecular target for bone regenerative therapies.

The physical interaction between SERINC3 and IL32γ defines a previously unrecognized axis in osteogenic regulation. Although IL32γ is well established as a multifunctional cytokine in immunity and cancer [39–41], our co-immunoprecipitation and rescue experiments reveal its essential role in SERINC3-mediated osteogenesis. This partnership may underlie the protective effects of IL32γ against bone loss [42], with SERINC3 potentially stabilizing IL32γ signaling through membrane incorporation. Considering the fact that SERINC3 is a potential regulator of phospholipid metabolism, SERINC3 might affect the IL32 secretion by altering membrane phospholipid composition. Structural studies using truncated mutants will be required to map the precise interaction domains governing this novel protein complex.

The SERINC3-IL32γ complex orchestrates a sophisticated metabolic program essential for osteogenesis through two interconnected pathways. First, SERINC3-OE potently activates AMPK phosphorylation and enhances autophagic flux, whereas pharmacological blockade of either AMPK (with Compound C) or autophagy (with bafilomycin A1) completely abolishes its osteogenic effects. These results position SERINC3 as a novel upstream regulator of the established autophagy-osteogenesis axis [40, 41], unveiling a previously unrecognized metabolic checkpoint in bone formation. Second, SERINC3 drives mitochondrial reprogramming by selectively upregulating key TCA cycle enzymes (ACO1, DLAT, and SDHA), thereby elevating oxidative phosphorylation and ATP production. The ability of electron transport chain modulators (CoQ10 and rotenone) to respectively rescue or inhibit SERINC3-mediated osteogenesis provides compelling evidence that mitochondrial metabolism serves as a critical effector mechanism downstream of SERINC3 signaling.

These mechanistic insights point toward promising therapeutic strategies. The superior bone regeneration capacity of SERINC3-engineered BMSC sheets in critical-sized defects highlights their potential for cell-based approaches in skeletal repair. Furthermore, the identification of the druggable AMPK-TCA axis opens the door to repurposing existing metabolic modulators, such as metformin, to enhance bone healing through pharmacological means.

Several key questions emerge from these findings. First, while our data establish that the physical interaction between SERINC3 and IL-32 is essential for AMPK activation, the precise downstream signaling mechanism remains to be fully elucidated. One possibility is that the SERINC3–IL-32 complex stabilizes or facilitates the recruitment of a canonical IL-32 receptor (e.g., integrin αVβ3 or PAR1), thereby promoting receptor-proximal signaling events. Alternatively, SERINC3 may act as a scaffold that directly assembles intracellular signaling molecules, such as AMPK kinases, independent of a conventional receptor. Distinguishing between these models will require future structural studies and mutagenesis-based dissection of the signaling complex. Nonetheless, our current findings identify the SERINC3–IL-32 interaction as a critical upstream node controlling AMPK phosphorylation.

Second, given SERINC3’s established role in viral particle incorporation [20, 37, 38] and the growing recognition of extracellular vesicles in osteogenesis [43, 44], it remains to be determined whether the SERINC3-IL32γ complex influences vesicle biogenesis or cargo selection. Third, the functional significance of specific TCA cycle enzymes (e.g., SDHA) remains unclear: do they act as direct mediators of osteogenic differentiation, or merely serve as bystanders in the broader metabolic rewiring process? The development of small molecules that specifically enhance the interaction between SERINC3 and IL32γ could provide a novel therapeutic strategy for bone regeneration and warrant further investigation. Furthermore, osteoclastic differentiation is a critical factor in bone homeostasis [45]. However, the regulatory role of SERINC3 in osteoclastogenesis warrants further investigation. Finally, Serinc3 knockout may exert systemic metabolic or developmental effects that indirectly influence the bone phenotype. Serinc3 conditional knockout in MSCs is therefore necessary to confirm the role of SEINC3-regulated bone formation in vivo.

Conclusions

Through integrated molecular, cellular, and in vivo analyses, we demonstrate that SERINC3 orchestrates bone formation by activating IL32γ-dependent AMPK-autophagy signaling and mitochondrial bioenergetics. These discoveries not only advance our fundamental understanding of osteogenic regulation but also establish SERINC3 as a promising target for developing innovative bone regenerative strategies. Future studies should investigate SERINC3’s role in vesicular trafficking and optimize pharmacological approaches to modulate its activity.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary Material 1. (5.2MB, pdf)
Supplementary Material 2. (53MB, doc)

Acknowledgements

The authors declare that they have not use AI-generated work in this manuscript.

Abbreviations

BMSCs

Bone marrow mesenchymal stem cells

SHED

Human exfoliated deciduous teeth

KO

Knockout

BV/TV

Bone volume/total volume

BMD

Bone mineral density

Tb.N

Trabecular number

Tb.Th

Trabecular thickness

Tb.Sp

Trabecular separation

MSCs

Mesenchymal stem cells

SERINC

Serine incorporator

PS

Phosphatidylserine

SP

Sphingolipids

hOBs

Human calvarial osteoblasts

ALP

Alkaline phosphatase

OE

Overexpression

KD

Knockdown

OD

Optical density

FCCP

Carbonyl cyanide-4-trifluoromethoxyphenylhydrazone

TEM

Transmission electron microscope

TCA

Tricarboxylic acid

Author contributions

Conceptualization, J.L., Z.Z., J.P., L.W., H.Z. R.J.; Formal analysis, Z.Z., T.X., J.P., L.W.; Funding J.L., L.W., Z.Z., H.Z.; Investigation, Z.Z., T.X., S.X., J.L., S.Y., Z.F., W.X., H.Z.; Supervision, J.L., L.W., H.Z.; Writing-original draft preparation, Z.Z, T.X.; Writing-review and editing, J.L., J.P., L.W, H.Z. All authors have read and agreed to the prepared version of the manuscript.

Funding

This work was supported by GuangDong Basic and Applied Basic Research Foundation (2023A1515030055), the Science and Technology Program of Guangzhou (202201020116), First-class major construction project (02-408-2501-2142), and Key Technologies R&D Program of Dongguan (20221200300072).

Data availability

The RNA sequencing data have been deposited in the Genome Sequence Archive (GSA) for the human database (HRA011989) (HTTPS://NGDC.CNCB.AC.CN/SEARCH/SPECIFIC? DB=HRA&Q=HRA011989). The datasets supporting the conclusions of this article are available in the Figshare repository, with the identifier https://doi.org/10.6084/m9.figshare.32298078.

Declarations

Ethics approval and consent to participate

The study exploring the osteogenic differentiation mechanism of stem cells from human exfoliated deciduous teeth complies with the Helsinki Declaration and was approved by the Ethics Committee of the School and Hospital of Stomatology, Guangzhou Medical University (Approval No.: LCYJ2021017; Date: March 30, 2021). Written informed consent was obtained from the parents of all participants. The animal experiments that evaluate regenerative therapy for bone defects, periodontitis, and skin defects via transgenic animal models and gene therapy adhere to the International Council for Laboratory Animal Science (ICLAS) guidelines and were approved by the Laboratory Animal Ethics Committee of Guangdong Huawei Testing Co., LTD (Approval No.: 202202002; Date: February 26, 2022). The original source (OriCell) has confirmed that there was initial ethical approval for the collection of human cells and that the donors had signed informed consent. The original source (BMSCs from Sciencell) has confirmed that there was initial ethical approval for the collection of human cells and that the donors had signed informed consent.

Consent for publication

All the authors have reviewed the manuscript and approve it for publication.

Competing interests

The authors declare no competing interests.

Footnotes

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Zhichao Zheng, Tianru Xu and Janak L. Pathak have contributed equally and share the first authorship.

Contributor Information

Huade Zheng, Email: hdzheng@scut.edu.cn.

Lihong Wu, Email: wcanhong@gzhmu.edu.cn.

Jiang Li, Email: ljiang@gzhmu.edu.cn.

References

  • 1.Majidinia M, Sadeghpour A, Yousefi B. The roles of signaling pathways in bone repair and regeneration. J Cell Physiol. 2018;233(4):2937–48. [DOI] [PubMed] [Google Scholar]
  • 2.Komatsu DE, Warden SJ. The control of fracture healing and its therapeutic targeting: improving upon nature. J Cell Biochem. 2010;109(2):302–11. [DOI] [PubMed] [Google Scholar]
  • 3.El-Rashidy AA, Roether JA, Harhaus L, Kneser U, Boccaccini AR. Regenerating bone with bioactive glass scaffolds: A review of in vivo studies in bone defect models. Acta Biomater. 2017;62:1–28. [DOI] [PubMed] [Google Scholar]
  • 4.Maruyama M, Rhee C, Utsunomiya T, Zhang N, Ueno M, Yao Z, et al. Modulation of the Inflammatory Response and Bone Healing. Front Endocrinol (Lausanne). 2020;11:386. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5.Barrett J, Craddock C. Bone marrow transplantation in the United Kingdom - past, present and future. Br J Haematol. 2020;191(4):612–6. [DOI] [PubMed] [Google Scholar]
  • 6.Wang W, Yeung KWK. Bone grafts and biomaterials substitutes for bone defect repair: A review. Bioact Mater. 2017;2(4):224–47. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7.Bai L, Zhou D, Li G, Liu J, Chen X, Su J. Engineering bone/cartilage organoids: strategy, progress, and application. Bone Res. 2024;12(1):66. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8.Zhou J, Shi Y. Mesenchymal stem/stromal cells (MSCs): origin, immune regulation, and clinical applications. Cell Mol Immunol. 2023;20(6):555–7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9.Alsaleem SA, Alkhairi MAY, Alzahrani MAA, Alwadai MI, Alqahtani SSA, Alaseri YFY, et al. Challenges and Barriers Toward Medical Research Among Medical and Dental Students at King Khalid University, Abha, Kingdom of Saudi Arabia. Front Public Health. 2021;9:706778. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.Liao Y, Fang Y, Zhu H, Huang Y, Zou G, Dai B, et al. Concentrated Growth Factors Promote hBMSCs Osteogenic Differentiation in a Co-Culture System With HUVECs. Front Bioeng Biotechnol. 2022;10:837295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11.Zhang Y, Feng X, Zheng B, Liu Y. Regulation and mechanistic insights into tensile strain in mesenchymal stem cell osteogenic differentiation. Bone. 2024;187:117197. [DOI] [PubMed] [Google Scholar]
  • 12.Inuzuka M, Hayakawa M, Ingi T. Serinc, an activity-regulated protein family, incorporates serine into membrane lipid synthesis. J Biol Chem. 2005;280(42):35776–83. [DOI] [PubMed] [Google Scholar]
  • 13.Stegen S, Loopmans S, Stockmans I, Moermans K, Carmeliet P, Carmeliet G. De novo serine synthesis regulates chondrocyte proliferation during bone development and repair. Bone Res. 2022;10(1):14. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14.Grewe JM, Knapstein P-R, Donat A, Jiang S, Smit DJ, Xie W, et al. The role of sphingosine-1-phosphate in bone remodeling and osteoporosis. Bone Res. 2022;10(1):34. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 15.Xu C, Zheng Z, Fang L, Zhao N, Lin Z, Liang T, et al. Phosphatidylserine enhances osteogenic differentiation in human mesenchymal stem cells via ERK signal pathways. Mater Sci Eng C Mater Biol Appl. 2013;33(3):1783–8. [DOI] [PubMed] [Google Scholar]
  • 16.Mäkitie O, Zillikens MC. Early-Onset Osteoporosis. Calcif Tissue Int. 2022;110(5):546–61. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17.Zhang M, Yu T, Li J, Yan H, Lyu L, Yu Y, et al. Matrix Metalloproteinase-Responsive Hydrogel with On-Demand Release of Phosphatidylserine Promotes Bone Regeneration Through Immunomodulation. Adv Sci (Weinh). 2024;11(20):e2306924. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18.Xu C, Su P, Chen X, Meng Y, Yu W, Xiang AP, et al. Biocompatibility and osteogenesis of biomimetic Bioglass-Collagen-Phosphatidylserine composite scaffolds for bone tissue engineering. Biomaterials. 2011;32(4):1051–8. [DOI] [PubMed] [Google Scholar]
  • 19.Li S, Lu X, Chai Q, Huang B, Dai S, Wang P, et al. Engineered Niobium Carbide MXenzyme-Integrated Self-Adaptive Coatings Inhibiting Periprosthetic Osteolysis by Orchestrating Osteogenesis-Osteoclastogenesis Balance. ACS Appl Mater Interfaces. 2024;16(23):29805–22. [DOI] [PubMed] [Google Scholar]
  • 20.Leonhardt SA, Purdy MD, Grover JR, Yang Z, Poulos S, McIntire WE, et al. Antiviral HIV-1 SERINC restriction factors disrupt virus membrane asymmetry. Nat Commun. 2023;14(1):4368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21.Xu S, Zheng Z, Pathak JL, Cheng H, Zhou Z, Chen Y, et al. The Emerging Role of the Serine Incorporator Protein Family in Regulating Viral Infection. Front Cell Dev Biol. 2022;10:856468. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 22.Li S, Ahmad I, Shi J, Wang B, Yu C, Zhang L, et al. Murine Leukemia Virus Glycosylated Gag Reduces Murine SERINC5 Protein Expression at Steady-State Levels via the Endosome/Lysosome Pathway to Counteract SERINC5 Antiretroviral Activity. J Virol. 2019;93(2):10–128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23.Timilsina U, Umthong S, Ivey EB, Waxman B, Stavrou S. SARS-CoV-2 ORF7a potently inhibits the antiviral effect of the host factor SERINC5. Nat Commun. 2022;13(1):2935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24.Zeng Y, Xiao D, He H, He J, Pan H, Yang W, et al. SERINC2-knockdown inhibits proliferation, migration and invasion in lung adenocarcinoma. Oncol Lett. 2018;16(5):5916–22. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 25.Jin Q, Feng J, Yan Y, Kuang Y. Prognostic and immunological role of adaptor related protein complex 3 subunit mu2 in colon cancer. Sci Rep. 2024;14(1):483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 26.Soleimani M, Nadri S. A protocol for isolation and culture of mesenchymal stem cells from mouse bone marrow. Nat Protoc. 2009;4(1):102–6. [DOI] [PubMed] [Google Scholar]
  • 27.Moussa FM, Cook BP, Sondag GR, DeSanto M, Obri MS, McDermott SE, et al. The role of miR-150 regulates bone cell differentiation and function. Bone. 2021;145:115470. [DOI] [PubMed] [Google Scholar]
  • 28.Zheng Z, Wu L, Li Z, Tang R, Li H, Huang Y, et al. Mir155 regulates osteogenesis and bone mass phenotype via targeting S1pr1 gene. Elife. 2023;12:e77742. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 29.Reyes R, Rodríguez JA, Orbe J, Arnau MR, Évora C, Delgado A. Combined sustained release of BMP2 and MMP10 accelerates bone formation and mineralization of calvaria critical size defect in mice. Drug Deliv. 2018;25(1):750–6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 30.Su P, Tian Y, Yang C, Ma X, Wang X, Pei J, et al. Mesenchymal Stem Cell Migration during Bone Formation and Bone Diseases Therapy. Int J Mol Sci. 2018;19(8):2343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 31.Bressan C, Saghatelyan A. AMPK-induced autophagy as a key regulator of cell migration. Autophagy. 2021;17(3):828–9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 32.Mihaylova MM, Shaw RJ. The AMPK signalling pathway coordinates cell growth, autophagy and metabolism. Nat Cell Biol. 2011;13(9):1016–23. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 33.Herzig S, Shaw RJ. AMPK: guardian of metabolism and mitochondrial homeostasis. Nat Rev Mol Cell Biol. 2018;19(2):121–35. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 34.Lee DH, Hong JE, Yun HM, Hwang CJ, Park JH, Han SB, et al. Interleukin-32beta ameliorates metabolic disorder and liver damage in mice fed high-fat diet. Obes (Silver Spring). 2015;23(3):615–22. [DOI] [PubMed] [Google Scholar]
  • 35.Zhao J, Chen A, Wang R, Qiu D, Chen H, Li J, et al. Bmi-1 Epigenetically Orchestrates Osteogenic and Adipogenic Differentiation of Bone Marrow Mesenchymal Stem Cells to Delay Bone Aging. Adv Sci (Weinh). 2024;11(46):e2404518. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 36.Yu S, Du J, Zhang Q, Li Z, Ge S, Ma B. Mitochondria-Targeted Polyphenol‐Cysteine Nanoparticles Regulating AMPK‐Mediated Mitochondrial Homeostasis for Enhanced Bone Regeneration. Adv Funct Mater. 2024;34:2402463. [Google Scholar]
  • 37.Chande A, Cuccurullo EC, Rosa A, Ziglio S, Carpenter S, Pizzato M. S2 from equine infectious anemia virus is an infectivity factor which counteracts the retroviral inhibitors SERINC5 and SERINC3. Proc Natl Acad Sci U S A. 2016;113(46):13197–202. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 38.Peng Y, Zong Y, Wang D, Chen J, Chen Z-S, Peng F, et al. Current drugs for HIV-1: from challenges to potential in HIV/AIDS. Front Pharmacol. 2023;14:1294966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 39.Wallimann A, Schenk M. IL-32 as a potential biomarker and therapeutic target in skin inflammation. Front Immunol. 2023;14:1264236. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 40.Sloot YJE, Smit JW, Joosten LAB, Netea-Maier RT. Insights into the role of IL-32 in cancer. Semin Immunol. 2018;38:24–32. [DOI] [PubMed] [Google Scholar]
  • 41.Kang J-W, Park YS, Lee DH, Kim MS, Bak Y, Ham SY, et al. Interaction network mapping among IL-32 isoforms. Biochimie. 2014;101:248–51. [DOI] [PubMed] [Google Scholar]
  • 42.Lee E-J, Kim S-M, Choi B, Kim E-Y, Chung Y-H, Lee E-J, et al. Interleukin-32 Gamma Stimulates Bone Formation by Increasing miR-29a in Osteoblastic Cells and Prevents the Development of Osteoporosis. Sci Rep. 2017;7:40240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 43.Wang D, Guo Y, Heng BC, Zhang X, Wei Y, He Y, et al. Cell membrane vesicles derived from hBMSCs and hUVECs enhance bone regeneration. Bone Res. 2024;12(1):23. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 44.Jiao Y, Liu Y, Du J, Xu J, Luo Z, Liu Y, et al. Advances in the Study of Extracellular Vesicles for Bone Regeneration. Int J Mol Sci. 2024;25(6):3480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 45.Sun XY, Wu T, Chen SH, Zhao ZY, Jia RW, Ma J, et al. OTUD1 inhibits osteoclast differentiation and osteoclastic bone loss through deubiquitinating and stabilizing PRDX1. Theranostics. 2025;15(14):6719–36. [DOI] [PMC free article] [PubMed] [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

Supplementary Material 1. (5.2MB, pdf)
Supplementary Material 2. (53MB, doc)

Data Availability Statement

The RNA sequencing data have been deposited in the Genome Sequence Archive (GSA) for the human database (HRA011989) (HTTPS://NGDC.CNCB.AC.CN/SEARCH/SPECIFIC? DB=HRA&Q=HRA011989). The datasets supporting the conclusions of this article are available in the Figshare repository, with the identifier https://doi.org/10.6084/m9.figshare.32298078.

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