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. 2022 Jan;9(1):e2104128.
doi: 10.1002/advs.202104128. Epub 2021 Nov 5.

Precise Diabetic Wound Therapy: PLS Nanospheres Eliminate Senescent Cells via DPP4 Targeting and PARP1 Activation

Renliang Zhao  1 , Xiangyun Jin  2 , Ang Li  1 , Bitong Xu  3 , Yifan Shen  1 , Wei Wang  1 , Jinghuan Huang  1 , Yadong Zhang  3 , Xiaolin Li  1
Affiliations

Precise Diabetic Wound Therapy: PLS Nanospheres Eliminate Senescent Cells via DPP4 Targeting and PARP1 Activation

Renliang Zhao et al. Adv Sci (Weinh). 2022 Jan.

Abstract

Diabetic ulcers, a difficult problem faced by clinicians, are strongly associated with an increase in cellular senescence. Few empirical studies have focused on exploring a targeted strategy to cure diabetic wounds by eliminating senescent fibroblasts (SFs) and reducing side effects. In this study, poly-l-lysine/sodium alginate (PLS) is modified with talabostat (PT100) and encapsulates a PARP1 plasmid (PARP1@PLS-PT100) for delivery to target the dipeptidyl peptidase 4 (DPP4) receptor and eliminate SFs. PARP1@PLS-PT100 releases encapsulated plasmids, displaying high selectivity for SFs over normal fibroblasts by targeting the DPP4 receptor, decreasing senescence-associated secretory phenotypes (SASPs), and stimulating the secretion of anti-inflammatory factors. Furthermore, the increased apoptosis of SFs and the disappearance of cellular senescence alleviates SASPs, accelerates re-epithelialization and collagen deposition, and significantly induces macrophage M2 polarization, which mediates tissue repair and the inflammatory response. This innovative strategy has revealed the previously undefined role of PARP1@PLS-PT100 in promoting diabetic wound healing, suggesting its therapeutic potential in refractory wound repair.

Keywords: DPP4 receptor; diabetic wound healing; nanospheres; selective targeting; senescence.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
A,B) Schematic illustration of the encapsulation of PARP1@PLS‐PT100 nanospheres; C) NMR analysis confirmed successful sodium alginate scion grafting of PT100; D) FTIR spectrum results showed characteristic bands of SA‐DA‐PT100; G) the morphology of a graded concentration ratio of PLL and SA complex and the E) complex were measured for particle size by NTA. PARP1 encapsulated PLS‐PT100 nanospheres were detected with F) NTA and H)TEM. (n = 3 images per group.)
Figure 2
Figure 2
A) GFP expression was indicated by green fluorescence in SFs after transfecting with liposomes, PLS, PLS@PT100. B) The relative PARP1 gene expression in SFs with the indicated treatment and the C) expression of the protein. (n = 3 per group; * p < 0.05, ns, p > 0.05.)
Figure 3
Figure 3
A) Proliferating human foreskin fibroblasts (HFF‐1) and B) senescent fibroblasts (SFs) were incubated with a series of nanospheres. The expression of SASPs: C) IL‐6; D: IL‐1α; E: IFN‐γ; F) TNF‐α) and anti‐inflammatory factors: G) IL‐3; H) G‐CSF was detected in HFF‐1 cells, which were incubated with the supernatant collected from SFs after treatment with PBS, PLS, PLS‐PT100, PARP1, PARP1@PLS, PARP1@PLS‐PT100 nanocarriers. Selective expression of DPP4 receptor in SFs compared with I) normal HFF‐1, J) quantitative evaluation of the DPP4 positive cell ratio. K) The immunofluorescence staining images indicated that PT100 modified nanospheres could target SFs, and L) the quantitative results were calculated. (n = 3 per group; * p < 0.05, ns, p > 0.05).
Figure 4
Figure 4
A) The expression of P16INK4a was measured ( and the C) number P16INK4a of positive cells was also calculated. B) The immunofluorescence staining images of α‐SMA on HFF‐1 cells with the treatment of SFs’ supernatant and the D) mean fluorescence intensity (MFI) were calculated. (n = 3 per group; * p < 0.05, ns, p > 0.05).
Figure 5
Figure 5
A) TUNEL fluorescence staining of wound tissues (. Staining of senescence‐associated B) b‐galactosidase (SA‐b‐gal) and the D) SA‐b‐gal‐positive area were calculated. C) Immunohistochemistry staining of Ki67 in wound sections at day 12 and day 21 after surgery and E) quantitative evaluation of the Ki67‐positive cell ratio. F) The expression of PARP1 and AIF in wound tissue on day 12 after surgery. (n = 3 per group; * p < 0.05, ns, p > 0.05).
Figure 6
Figure 6
A) Immunofluorescence staining of CD206 and B) quantitative evaluation of CD206‐positive cells. E,F) Flow cytometry of wound tissue at days 12 and 21 after surgery, and the C,D) analysis of flow cytometry were calculated. The relative SASP (IL‐1α, IL‐6, TNF‐α, and IFN‐γ) and anti‐inflammatory factor (IL‐3, CSF) gene expression in rat wound tissues at G) day 12 and H) day 21, (n = 3 per group; * p < 0.05, ns, p > 0.05).
Figure 7
Figure 7
A) Gross photographs of wound closure and B) simulation plots of wound closure. C) H&E staining of the wound indicated the healing situation on days 12 and 21. The quantitative analysis of D) wound closureand E) H&E staining. (n = 3 per group; * p < 0.05, ns, p > 0.05.)
Figure 8
Figure 8
A) Masson staining and B) Sirius red staining indicated collagen deposition and alignment. D) The analysis of Sirius red staining indicated the ratio of collagen I (yellow) / collagen III (green). C) The immunofluorescence staining images of α‐SMA and the E) quantitative analysis of α‐SMA positive area rate. (n = 3 per group; * p < 0.05, ns, p > 0.05.)
Scheme 1
Scheme 1
Schematic illustration of therapeutic nanocarriers releasing PARP1 pDNA for wound healing. a) Preparation of PARP1@PLS‐PT100 nanocarriers. b) senescence is the significant source of inflammation and persistent continuing inflammatory microenvironment, and inflammation accelerates the progression of senescence. c) Sustained release of pDNA with the injection of nanospheres promoted the expression of AIF and its release and leading to the senescent fibroblast apoptosis d) Regeneration of wound Senescenct wound healing process tissue by the composited with PARP1@PLS‐PT100 therapeutic nanospheres.
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References

    1. Armstrong D. G., Boulton A. J. M., Bus S. A., N. Engl. J. Med. 2017, 376, 2367. - PubMed
    1. Falanga V., Lance 2005, 366, 1736. - PubMed
    1. Dorschner R. A., Lee J., Cohen O., Costantini T., Baird A., Eliceiri B. P., Sci. Adv. 2020, 6, eaay0518. - PMC - PubMed
    1. Wong S., Demers M., Martinod K., Gallant M., Wang Y., Goldfine A., Kahn C., Wagner D., Nat. Med. 2015, 21, 815. - PMC - PubMed
    1. Wu P. Y., Huang C. C., Chu Y., Huang Y. H., Lin P., Liu Y. H., Wen K. C., Lin C. Y., Hsu M. C., Chiang H. M., Int. J. Mol. Sci. 2017, 18. 10.3390/ijms18040782. - DOI - PMC - PubMed

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