BMSC derived exosomes accelerate diabetic wound healing via circ-Snhg11 delivery

doi:10.21203/rs.3.rs-1638618/v1

Download PDF

Research Article

BMSC derived exosomes accelerate diabetic wound healing via circ-Snhg11 delivery

https://doi.org/10.21203/rs.3.rs-1638618/v1

This work is licensed under a CC BY 4.0 License

Version 1

posted

You are reading this latest preprint version

Background: Exosome (Exo) secreted by bone mesenchymal stem cells (BMSCs) enhances cutaneous wound healing in mouse with diabetes mellitus (DM). However, the mechanism remains unclear.

Methods: Next-generation sequencing (NGS) was applied to study abnormal circular RNA (circRNA) expressions in ulcer tissues with or without Exo treatment. Bioinformatics and luciferase outputs were employed to verify interactional correlations amongst RNAs. EPCs cells were utilized to study the ROS under high glucose conditions by immunofluorescence. A DM ulceration mouse model was established and the therapeutic Exo effect was investigated through immunofluorescence.

Results: Data showcased that Exo treatment significantly promoted cutaneous wound healing in mice with DM. NGS discovered that circ-Snhg11 functioned in Exo-mediated tissues repair. circ-Snhg11 downregulation reduced therapeutic effects regarding Exo upon wound healing in diabetic mice. Bioinformatics and luciferase reporting outcomes verified that both miR-144-3p and SLC7A11 were circ-Snhg11 downstream targets. SLC7A11 downregulation or miR-144-3p overexpression reversed protective effect regarding circ-Snhg11 on EPCs after exposure to a high glucose microenvironment. circ-Snhg11 upregulation increased the therapeutic Exo effect on wound healing in diabetic mice by stimulating angiogenesis and reducing GPX4-mediated ferroptosis. Furthermore, Exo circ-Snhg11 delivery promoted the miR-144-3p/SLC7A11/GPX4 signal.

Conclusions: In conclusion, circ-Snhg11 from BMSC Exo promoted the SLC7A11/GPX4-mediated anti-ferroptosis signal by sponging miR-144-3p. This promoted angiopoiesis and accelerated diabetic wound healing.

Exosomes

diabetic wound healing

bone mesenchymal stem cells

circ-Snhg11

EPCs

Diabetes mellitus (DM) is a chronic global trait. From epidemiological survey published by the WHO, diabetes prevalence amongst adults (20–79 years) in the world would increment from 6.4% in 2010 to 7.7% in 2030, so that the affected adult numbers would increment from 285 million to 439 million [1, 2]. Hyperglycemia-induced vascular damage might be a main factor causing serious diabetic complications such like diabetic foot ulcers [3, 4]. In this matter, it is super critical to discover new treatment methods for diabetic ulcers.

Recently, mesenchymal stem cells (MSCs) are elucidated to function importantly during tissue regeneration and wound repairing via accelerating wound closure by promoting angiogenesis and modulating the microenvironment [5]. Previous studies found that diabetic ferroptosis functions importantly to trigger inflammation in the diabetic wound [6]. Excessive expression regarding inflammatory factors disrupts angiogenesis [7, 8]. Many investigations revealed that exosomes (Exo) from MSCs function importantly to regulate organizational microenvironment [9, 10]. Circular RNA (circRNA) function importantly in Exo-mediated bioinformation regulation [11]. CircRNAs have an important function to regulate gene expressions indicating that they are candidate therapeutic and diagnostic markers [12].

Our previous study discovered that Exo from bone MSCs (BMSCs) accelerates wound healing in diabetic mice. Next-generation sequencing discovered that circ-Snhg11 expression was incremented after Exo treatment in the ulcer site. lowered in diabetic mice. The mechanistic study discovered that circ-Snhg11 promotion regarding SLC7A11/GPX4 expression mediated an anti-ferroptosis signal by sponging miR-144-3p, which promoted angiopoiesis and accelerated diabetic wound healing.

BMSC isolation, culturing and identification

Tibia and femur from BALB/c mice were extracted under sterile conditions to expose bone marrow cavity, which we rinsed with saline. The bone marrow filtrate was gained and centrifuged at 225 × g for five minutes. Supernatant was discarded and we resuspended cells in low-glucose (LG)-Dulbecco's modified Eagle's medium (DMEM) (LG-DMEM; HyClone, Logan) at 1×10⁶ cells per 100 μl. Cell suspension was added gradually to mice lymphocyte separation medium (Sigma-Aldrich) with 1:1 ratio and centrifuged at 1000×g for twenty minutes. Milky turbid mononuclear cell layer was obtained. Technician resuspended cells in LG-DMEM medium without FBS at 1×10⁶ cells per 100 μl, which were centrifuged at 225 × g for five minutes. Pelleted cells were gained. Technician resuspended cells in LG-DMEM complete medium containing 10% FBS under 5% CO₂ and saturated humidity at 37 °C. Culture medium was altered every 3d, which we subcultured with 1:3 ratio when cell confluency reached 80~90%. BMSCs at passages 3~4 were utilized for the following experiment. For phenotypic analysis, we used phycoerythrin (PE). The following marker expressions were studied: CD29, CD105, CD44, CD90, and von Willebrand factor (vWF). Immunoglobulin G-matched isotype served as internal control for every antibody.

Multilineage BMSC differentiation

To identify the BMSC multilineage differentiation capacity, we cultivated 3rd passage mouse BMSCs. For adipocyte differentiations, we cultured BMSCs in adipogenic differentiation medium. After 14 days, technician conducted oil red O staining to confirm adipocyte differentiation. For osteoblast differentiation, we cultured MSCs in osteogenic differentiation medium. After 3 weeks, we stained cells with alizarin red or ALP.

ADSC-Exo isolation and identification

Upon achieving 80~90% confluency, we rinsed BMSCs utilizing phosphate-buffered saline (PBS) and cultured them in EGM-2MV media without FBS and supplemented with 1× serum replacement solution (PeproTech, New Jersey, USA) for another 2d. Conditioned medium from BMSCs was gained and centrifuged at 300×g for ten minutes and 2000×g for 10 min to eliminate cellular debris and dead cells. Lastly, after centrifugation at 12, 000×g for 0.5 h. All processes were done at 4 °C. Exo protein content was defined employing Pierce BCA Protein Assay Kit (Thermo Fisher Scientific, MA, USA). BMSC-Exos were maintained at -80 °C or immediately utilized for downstream experiments. We applied transmission electron microscopy, NTA assay and western blotting to characterize Exos that collected.

Strand-specific NGS lib preparations

Our lab obtained total RNA from BMSC Exos or PBS-treated diabetic ulcer tissue utilizing TRIzol reagent (Invitrogen, Carlsbad, CA, USA). Technician used ~3μg total RNA from every specimen through VAHTS Total RNA-seq (H/M/R) Library Prep kits from Illumina (Vazyme Biotech Co., Ltd, Nanjing, China) to eliminate ribosomal RNA while retaining other RNAs such like ncRNAs and mRNAs. Our team treated RNA through 40 U RNase R (Epicenter) at 37 °C for 3 h, followed by TRIzol purification. RNA-seq libraries were prepared through KAPA Stranded RNA-Seq Library Prep kits (Roche, Basel, Switzerland), which we employed for NGS (Illumina HiSeq 4000 at Aksomics, Inc., Shanghai, China).

Bioinformatics analyses

Interactional correlations among RNAs were calculated via the website-based tool.

Cell culture and transfection

Our team obtained EPCs cells from ScienCell. We cultured cells in DMEM (HyClone) and 10% FBS (Gibco, Carlsbad, CA), which we stored in humidified incubator at 37 °C under 5% CO₂. SLC7A11 gene and circ-Snhg11 overexpression vector were constructed by inserting circ-Snhg11 cDNA into the pcDNA3.1 vector. miR-144-3p mimics and SLC7A11 siRNA were synthesized by Genepharma (Suzhou, China). Cell transfection was made through Lipofectamine 2000 (Invitrogen) following protocols.

RNA isolation and real-time PCR

Total RNA was obtained with TRIzol Reagent (Invitrogen), followed by cDNA synthesis utilizing TransScript All-in-One First-Strand cDNA Synthesis SuperMix (Transgen Biotech, Beijing, China), which we made following [24]. PCR was made utilizing Bio-Rad PCR instrument (Bio-Rad, Hercules, CA, U.S.A.) using 2× Taq PCR Master Mix (Solarbio, Beijing, China) following protocols. Fold changes were computed by relative quantification means from 2^−△△Ct method. The PCR primers are:

circ-Snhg11: forward 5′-GTTCTGTGATGGTTCCTC-3′ and reverse 5′-CAGCAGCGGAGTCCACG-3′; miR-144-3p: forward 5′-TCATGTAGTAGATATGACAT-3′ and reverse 5′-GGCAGGGTCCGAGGTATTC-3′; SLC7A11: forward 5′-ATACGCTGAGTGTGGTTTGC-3′ and reverse 5′-CTTCATCCACTTCCACAGCG-3′; GPX4: forward 5′-ATACGCTGAGTGTGGTTTGC-3′ and reverse 5′-CTTCATCCACTTCCACAGCG-3′; U6: forward 5′-CTCTCGCTTCGGCAGCACA-3′ and reverse 5′-ACGCTTCACGAATTTGCGT-3′; GAPDH forward 5′-GCAAGGATGCTGGCGTAATG-3′ and reverse 5′-TACGCGTAGGGGTTTGACAC-3′.

Dual-luciferase reporter assay

SLC7A11 and circ-Snhg11 3′UTR including binding sites that predicated for miR-144-3p were amplified by PCR. We inserted fragments into multiple cloning sites in pMIR-REPORT luciferase miRNA expression reporter vector (Ambion, Austin, U.S.A.). We co-transfected EPCs cells with 0.1 μg luciferase reporter vectors including WT or MUT types of the SLC7A11 or circ-Snhg11 3′UTR. The same to miR-144-3p mimic or miR-control using Lipofectamine 2000 (Invitrogen). Relative luciferase activity was computed by normalizing firefly luminescence to Renilla luminescence employing Dual-Luciferase Reporter Assay System (Promega, Madison, WI, U.S.A.) following protocols at 2d post-transfection.

Reactive oxygen species (ROS) analysis

Our lab captured ROS production in EPCs or scar skin employing 2',7'-dichlorofluorescin diacetate (DCF-DA, Molecular Probes, Eugene, OR, USA). Technician seeded EPCs (1×10⁶ cells) in 100-mm culture dishes that pretreated through DAPI for 0.5~1 h and incubated with DCF-DA (20 mM) for 10 min at 37 ºC in dark. After 10 min incubation under normoxia or hypoxia, we examined cell fluorescence images to detect their ROS productions through fluorescence microscope (ECLIPSE E600, Nikon, Tokyo, Japan). Fluorescence intensity regarding DCF-DA was detected and computed by flow cytometry (BD Biosciences, San Jose, CA, USA). For tissues, the ROS were determined immediately after skin was excised from the mice.

Tubule formation assay

We assayed in vitro neovascularization in fibrin matrices. Following treatments, serum-starved EPCs in endothelial basal medium were seeded onto plates coated with Matrigel (10⁵ cells / well into 6 wells) (BD Biosciences, Franklin Lakes, NJ, USA) and incubated at 37 °C for 0.5 d. Technician photographed tubular structures that generated in Matrigel by phase-contrast microscopy. Lengths of the newly formed tubes in 10 randomly selected fields in each well were detected.

Diabetic wound induction

We applied BALB/c mice (male) for diabetes induction through single intraperitoneal injection w.r.t. 60 mg/kg streptozotocin (STZ) dissolved in 0.1 M citrate buffer (pH=4.5). At 3d post STZ administration, diabetes was verified by fasting blood glucose level measurements regarding blood obtained from tail vein. Mice with fasting blood glucose levels >250 mg/dL were regarded as diabetic, which we kept for 1 month and for next-step studies once the blood glucose had stabilized. Following diabetes confirmations, rats were anesthetized by intramuscular injection of a ketamine hydrochloride and xylazine cocktail at 80 and 10 mg/kg, respectively. Hair was erased from dorsal leg region and the area sterilized with povidone iodine solution. A 4 mm full-thickness excisional wound was made utilizing sterile biopsy punch. Our lab randomly allocated mice for 200 μg subcutaneous injection BMSC Exos in 100 μL PBS or the equal volume of PBS alone at 4 sites around the wound (25 μL / site). We euthanized mice after 15 d and harvested skin specimens for histopathological validations.

Immunohistochemical data

Our team fixed tissue samples with 4% paraformaldehyde solution, which we embedded in paraffin. We obtained sections, which we cultured with primary antibodies against CD31 or GPX4 over the night at 4 °C with secondary antibodies (Abcam) for 1h at 37 °C. We stained sections with 3,3-diaminobenzidine, which we counter-stained via hematoxylin.

Statistics analyses

Continuous variables were represented by means ± SD and one-way variance analysis was utilized to compare by GraphPad Prism (GraphPad, La Jolla, USA). P ≤ 0.05 informed statistical significance.

BMSC and BMSC-Exo identification

Present study isolated BMSCs, which have a classical cobblestone-like morphology (Fig. 1A). Immunofluorescence staining was positive for CD44, CD90, CD105 and CD29 expression, while negative for endothelial marker vWF (Fig. 1B-G). Results concerning ALP and oil red O staining verified the osteoblast and adipocyte differentiation potencies of BMSCs, respectively. (Fig. 1H and 1I). BMSC-Exos displayed a sphere- or cup-shaped morphology in transmission electron micrographs (Fig. 1J), which was similar to the Exos in [13]. NTA showed that the mean Exo diameter was approximately 100 nm (Fig. 1K). The Exo marker protein expressions such like CD63 and CD81 was shown with western blotting (Fig. 1L). The outputs informed that nanoparticles were Exos.

BMSC Exo effects on wound healing in diabetic mice.

Present study advised that BMSC Exo treatment accelerated diabetic wound healing (Fig. 2A and 2B). Immunofluorescence for ROS detection showed that Exo treatment decreased the ROS level in ulcer site tissues (Fig. 2C and 2D). Immunofluorescence for GPX4 detection showed that Exo treatment decreased its expression in ulcer site tissues (Fig. 2C and 2D). This suggested that Exo treatment decreased high glucose (HG)-induced ferroptosis in diabetic mice. Immunofluorescence for CD31 detection showed that Exo treatment increased its expression in ulcer site tissues (Fig. 2E and 2F). This suggested that Exo treatment reversed HG-induced disfunction of angiogenesis in ulcer site tissues of diabetic mice.

Mmu_circ_0001100 (circ-Snhg11) functions importantly in BMSC Exo-mediated wound repair from diabetic mice.

An accumulation study found that circRNA functions importantly in microenvironmental regulation. This study also discovered that Exo treatment resulted in abnormal circRNA expression by high-throughput sequencing in ulcer site tissues of diabetic mice (Fig. 3A). RT-qPCR detection showed that compared with the PBS treatment group, Exo treatment resulted in high mmu_circ_0001100 (circ-Snhg11) expression (Fig. 3B). RT-qPCR data showcased that circ-Snhg11 expression decreased in DM serum (Fig. 3C). Current investigation discovered that circ-Snhg11 silencing decreased the protective effect of Exo in accelerating diabetic wound healing (Fig. 3D and 3E). Immunofluorescence for ROS detection showed that circ-Snhg11 silencing decreased restored the ROS level after Exo treatment in ulcer site tissues (Fig. 3F and 3G). Immunofluorescence for GPX4 detection showed that circ-Snhg11 silencing reversed Exo treatment-induced GPX4 expression in ulcer site tissues (Fig. 3H and 3I). Immunofluorescence for CD31 detection demonstrated that circ-Snhg11 silencing reversed Exo treatment-induced CD31 expression in ulcer site tissues (Fig. 3J and 3K). Together, this suggested that circ-Snhg11 functions importantly in BMSC Exo-mediated wound repair in diabetic mice.

SLC7A11 functions essentially in BMSC Exo-mediated wound repair from diabetic mice.

In this study, we also found that Exo treatment resulted in abnormal mRNA expression by high-throughput sequencing in ulcer site tissues of diabetic mice (Fig. 4A). RT-qPCR detection showed that compared with the PBS treatment group, Exo treatment resulted in high SLC7A11 expressions (Fig. 4B). RT-qPCR detection demonstrated that SLC7A11 expression was decremented in DM serum (Fig. 4C). Current study advised that circ-Snhg11 silencing decreased SLC7A11 expression in accelerating diabetic wound healing (Fig. 4D).

MiR-144-3p and SLC7A11 were circ-Snhg11 downstream targets.

Bioinformatics results advised that mmu_circ_0001100 is originated from Snhg11 gene exons that located at chr2:158208239–158208899. Snhg11 is 660 bp and spliced mature circRNA is 660 bp, so, our team also called mmu_circ_0001100 as circ-Snhg11 (Fig. 5A). RT-qPCR detection showed miRNA expression, which can interact with both circ-Snhg11 and the 3'UTR of SLC7A11. Outcomes showcased that only miR-144-3p expression decreased in ulcer site tissues post Exo treatment (Fig. 5B).

Bioinformatics outputs (http://starbase.sysu.edu.cn/) revealed that circ-Snhg11 might interact with miR-144-3p. A luciferase reporter data verified that miR-144-3p suppressed luciferase activity in WT yet not MUT cell lines (Fig. 5C-D), advising that miR-144-3p was circ-Snhg11 target.

Bioinformatics data found that SLC7A11 was miR-144-3p downstream target. To validate correlations between miR-144-3p and SLC7A11, MUT or WT 3'UTR-SLC7A11 sequences containing miR-144-3p binding sequence were inserted into luciferase reporter vector (Fig. 5E). Luciferase reporter vector was transfected to EPCs cells integrated with or not miR-144-3p mimic. Luciferase reporter analyses revealed that miR-144-3p suppressed luciferase activity in WT yet not MUT cells (Fig. 5F), telling that SLC7A11 was miR-144-3p target.

SLC7A11 downregulation or miR-144-3p overexpression reversed protective effect regarding circ-Snhg11 upon EPCs post exposure to HG microenvironment.

RT-qPCR data showcased that circ-Snhg11 expression incremented post transfection with circ-Snhg11 overexpression vector. However, treatment with miR-144-3p mimic or SLC7A11 silence vector (si-SLC7A11) did not affect circ-Snhg11 expression regarding EPCs cells (Fig. 6A), which advised that miR-144-3p along with SLC7A11 was circ-Snhg11 downstream targets. RT-qPCR results discovered that circ-Snhg11 overexpression decremented miR-144-3p expression. SLC7A11 silencing did not affect circ-Snhg11-induced miR-144-3p downregulation (Fig. 6B). This advised that miR-144-3p was circ-Snhg11 downstream. The results also demonstrated that SLC7A11 silencing decreased its expression. By contrast, miR-144-3p upregulation reversed reducing effect w.r.t. circ-Snhg11 upon SLC7A11 expression. Following transfection with si-SLC7A11, SLC7A11 expression was significantly decreased (Fig. 6C). This suggested that circ-Snhg11 promotes SLC7A11 expression by sponging miR-144-3p.

RT-qPCR detection showed that SLC7A11 downregulation or miR-144-3p overexpression reversed circ-Snhg11-induced GPX4 expression in EPCs after exposure to a HG microenvironment (Fig. 6D). Immunofluorescence detection showed that SLC7A11 downregulation or miR-144-3p overexpression restored ROS level in circ-Snhg11-overexpressing EPCs after exposure to a HG microenvironment (Fig. 6E and 6F). An angiogenic differentiation experiment confirmed that SLC7A11 downregulation or miR-144-3p overexpression reversed circ-Snhg11-induced EPC angiogenic differentiation after exposure to a HG microenvironment (Fig. 6G and 6H).

SLC7A11 downregulation or miR-144-3p overexpression reversed circ-Snhg11 protective effects mediated by Exo on wound healing.

In this study, we also found that SLC7A11 downregulation or miR-144-3p overexpression reversed Exo-circ-Snhg11 protective effects to accelerate diabetic wound healing (Fig. 7A and 7B). Immunofluorescence for ROS detection showed that SLC7A11 downregulation or miR-144-3p overexpression restored ROS level after Exo-circ-Snhg11 treatment in ulcer site tissues (Fig. 7C and 7D). Immunofluorescence detection for GPX4 (Fig. 7E and 7F) and CD31 (Fig. 7G and 7H) showed that SLC7A11 downregulation or miR-144-3p overexpression reversed promotional effects regarding Exo-circ-Snhg11 upon GPX4 and CD31. This suggested that SLC7A11 downregulation or miR-144-3p overexpression reversed protective effects regarding circ-Snhg11 mediated by Exo on wound healing.

Former research advised that BMSCs have potential therapeutic effects to accelerate diabetic wound healing [14]. BMSC-derived Exos display potential therapeutic effects on tissue regeneration and angiogenesis by delivering bioinformatic molecules [15]. In this study, we also found that pretreatment with Exo from BMSCs accelerated diabetic wound healing. The data showed that Exo treatment promoted angiogenesis and decreased the ROS level in the ulcer site. At the same time, we found that Exo treatment increased GPX4 expression in the ulcer site. This suggested that Exo treatment reduced GPX4-mediated ferroptosis and restored EPC function.

The accumulation study found that circRNA plays an important role in Exo-mediated bioinformatics [16, 17]. Present study advised that Exo treatment incremented mmu_circ_0001100 (circ-Snhg11) expression in the ulcer site by high-throughput sequencing. Circ-Snhg11 downregulation reduced treatment effects regarding Exo in diabetic wound healing by restoring the ROS level and inhibiting angiogenesis. We further found that Exo treatment increased SLC7A11 (the key cystine transporter) expression in ulcer site. Former investigations found that SLC7A11 gene belongs to solute transport family and encodes cystine/glutamate xCT transporter, which is an essential gene regulating “iron overload-ferroptosis” [18]. Therefore, SLC7A11 was shown to be a suppressor of ferroptosis, and its inhibition by distinct approaches is able to trigger ferroptosis [19, 20].

Bioinformatics and luciferase reporter analyses found that miR-144-3p interacted with SLC7A11 and circ-Snhg11 3'UTR. Circ-Snhg11 overexpression decreased miR-144-3p, but promoted SLC7A11 expression. Former research revealed that miR-144-3p expression inhibits EPC mobilization [21]. Present results revealed that SLC7A11 downregulation or miR-144-3p overexpression reversed protective effects regarding circ-Snhg11 upon EPCs after exposure to a HG microenvironment by inhibiting GPX4-mediated ferroptosis in in vitro and in vivo experiments. Ferroptosis-related characteristic alternations were discovered in diabetic wound models and ferroptosis suppression improved inflammatory infiltration regarding diabetic wounds [6].

Taken together, the present study found that BMSC-derived Exos accelerate diabetic wound healing by circ-Snhg11 delivery and inhibit HG-induced ferroptosis. Analysis of the mechanism found that circ-Snhg11 expression promotes the SLC7A11/GPX4-mediated anti-ferroptosis signal by sponging miR-144-3p, thereby promoting angiopoiesis and accelerating diabetic wound healing.

Acknowledgements

None.

Funding

This work was Supported by the Fundamental Research Funds for the Central Universities (0214-14380481), and the Project funded by China Postdoctoral Science Foundation (2020M670035ZX), and the Natural Science Foundation of Jiangsu Province (SBK2020040321), and the Postdoctoral Research Funding Program of Jiangsu Province (2020Z368), and the Nanjing special fund for health science and technology development (YKK21073), and the 2020 Jiangsu Province Shuangchuang Ph.D. Introducing Talent Project of Nan Hu.

Availability of data and materials

Raw data is available from the corresponding author upon reasonable request.

Authors’ contributions

TT, LC, and CW conceived and wrote the manuscript. XD and SY revised the paper. NC, HC, XL reviewed and edited the manuscript. All the authors read and approved the final manuscript.

Ethics approval and consent to participate

This animal experiment was approved by the The Affiliated Nanjing Drum Tower Hospital, Nanjing University Medical School.

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

Cho NH, Shaw JE, Karuranga S, Huang Y, da Rocha Fernandes JD, Ohlrogge AW, et al. IDF Diabetes Atlas: Global estimates of diabetes prevalence for 2017 and projections for 2045. Diabetes Res Clin Pract. 2018;138:271–81.
Yang J, Chen Z, Pan D, Li H, Shen J. Umbilical Cord-Derived Mesenchymal Stem Cell-Derived Exosomes Combined Pluronic F127 Hydrogel Promote Chronic Diabetic Wound Healing and Complete Skin Regeneration. Int J Nanomedicine. 2020;15:5911–26.
Lim JZ, Ng NS, Thomas C. Prevention and treatment of diabetic foot ulcers. J R Soc Med. 2017;110:104–9.
Everett E, Mathioudakis N. Update on management of diabetic foot ulcers. Ann N Y Acad Sci. 2018;1411:153–65.
Hu Y, Tao R, Chen L, Xiong Y, Xue H, Hu L, et al. Exosomes derived from pioglitazone-pretreated MSCs accelerate diabetic wound healing through enhancing angiogenesis. J Nanobiotechnol. 2021;19:150.
Li S, Li Y, Wu Z, Fang H. Diabetic ferroptosis plays an important role in triggering on inflammation in diabetic wound. Am J Physiol Endocrinol Metab. 2021;321:E509-E20.
Liu W, Yu M, Xie D, Wang L, Ye C, Zhu Q, et al. Melatonin-stimulated MSC-derived exosomes improve diabetic wound healing through regulating macrophage M1 and M2 polarization by targeting the PTEN/AKT pathway. Stem Cell Res Ther. 2020;11:259.
Kasuya A, Tokura Y. Attempts to accelerate wound healing. J Dermatol Sci. 2014;76:169–72.
Wang M, Wang C, Chen M, Xi Y, Cheng W, Mao C, et al. Efficient Angiogenesis-Based Diabetic Wound Healing/Skin Reconstruction through Bioactive Antibacterial Adhesive Ultraviolet Shielding Nanodressing with Exosome Release. ACS Nano. 2019;13:10279–93.
Yu M, Liu W, Li J, Lu J, Lu H, Jia W, et al. Exosomes derived from atorvastatin-pretreated MSC accelerate diabetic wound repair by enhancing angiogenesis via AKT/eNOS pathway. Stem Cell Res Ther. 2020;11:350.
Shi R, Jin Y, Hu W, Lian W, Cao C, Han S, et al. Exosomes derived from mmu_circ_0000250-modified adipose-derived mesenchymal stem cells promote wound healing in diabetic mice by inducing miR-128-3p/SIRT1-mediated autophagy. Am J Physiol Cell Physiol. 2020;318:C848-C56.
Zhang L, Zeng M, Tang F, Chen J, Cao D, Tang ZN. Circ-PNPT1 contributes to gestational diabetes mellitus (GDM) by regulating the function of trophoblast cells through miR-889-3p/PAK1 axis. Diabetol Metab Syndr. 2021;13:58.
Lu GD, Cheng P, Liu T, Wang Z. BMSC-Derived Exosomal miR-29a Promotes Angiogenesis and Osteogenesis. Front Cell Dev Biol. 2020;8:608521.
Chen S, Shi J, Zhang M, Chen Y, Wang X, Zhang L, et al. Mesenchymal stem cell-laden anti-inflammatory hydrogel enhances diabetic wound healing. Sci Rep. 2015;5:18104.
Pomatto M, Gai C, Negro F, Cedrino M, Grange C, Ceccotti E, et al. Differential Therapeutic Effect of Extracellular Vesicles Derived by Bone Marrow and Adipose Mesenchymal Stem Cells on Wound Healing of Diabetic Ulcers and Correlation to Their Cargoes. Int J Mol Sci. 2021;22.
Cao G, Meng X, Han X, Li J. Exosomes derived from circRNA Rtn4-modified BMSCs attenuate TNF-alpha-induced cytotoxicity and apoptosis in murine MC3T3-E1 cells by sponging miR-146a. Biosci Rep. 2020;40.
Fu M, Fang L, Xiang X, Fan X, Wu J, Wang J. Microarray analysis of circRNAs sequencing profile in exosomes derived from bone marrow mesenchymal stem cells in postmenopausal osteoporosis patients. J Clin Lab Anal. 2022;36:e23916.
Xie Y, Hou W, Song X, Yu Y, Huang J, Sun X, et al. Ferroptosis: process and function. Cell Death Differ. 2016;23:369–79.
Koppula P, Zhuang L, Gan B. Cystine transporter SLC7A11/xCT in cancer: ferroptosis, nutrient dependency, and cancer therapy. Protein Cell. 2021;12:599–620.
Zhang W, Sun Y, Bai L, Zhi L, Yang Y, Zhao Q, et al. RBMS1 regulates lung cancer ferroptosis through translational control of SLC7A11. J Clin Invest. 2021;131.
Liu Y, Xu J, Gu R, Li Z, Wang K, Qi Y, et al. Circulating exosomal miR-144-3p inhibits the mobilization of endothelial progenitor cells post myocardial infarction via regulating the MMP9 pathway. Aging. 2020;12:16294–303.

Download PDF

Version 1

posted

You are reading this latest preprint version

BMSC derived exosomes accelerate diabetic wound healing via circ-Snhg11 delivery

Status:

Version 1

Abstract

Figures

Introduction

Materials And Methods

Results

Discussion

Conclusion

Declarations

References

Status:

Version 1