Human Parathyroid Hormone Analog (3–34/29–34) promotes wound re-epithelialization through inducing keratinocyte migration and epithelial–mesenchymal transition via PTHR1-PI3K/AKT activation

Abstract

Background Re-epithelialization is important in the process of wound healing. Various methods have been identified to expedite the process, but their clinical application remains limited. While parathyroid hormone (PTH) has shown promising results in wound healing due to its role in promoting collagen deposition and cell migration, application is limited by its potentially inhibitive effects when being continuously and locally administrated. Herein, we developed a novel PTH analog, Human parathyroid hormone (hPTH) (3–34/29–34) (henceforth MY-1), by partially replacing and repeating the amino acid sequences of hPTH (1–34), and evaluated its effect on skin wound re-epithelialization. Methods CCK-8, colony formation unit assay, and Ki67 immunofluorescent staining were performed to evaluate the effect of MY-1 on HaCaT cell proliferation. Then, wound scratch assay, Transwell assay and lamellipodia staining were carried out to evaluate the effect of MY-1 on cell migration. Moreover, the epithelial–mesenchymal transition (EMT) markers were measured using qPCR and western blot analysis. For in-vivo drug delivery, gelatin methacryloyl (GelMA) hydrogel was employed to load the MY-1, with the physicochemical characteristics evaluated prior to its application in wound models. Then, MY-1’s role in wound healing was determined via acute skin wound models. Finally, the mechanism that MY-1 activated was also detected on HaCaT cells and in-vivo wound models. Results In-vitro, MY-1 accelerated the migration and EMT of HaCaT cells, while having little effect on cell proliferation. GelMA and MY-1-incorporated GelMA hydrogels showed similar physicochemical characteristics and were used in the in-vivo studies, where the results revealed that MY-1 led to a stronger re-epithelialization by inducing basal keratinocyte migration and EMT. Further studies on in-vivo wound models and in-vitro HaCaT cells revealed that MY-1 regulated cell migration and EMT through activating PI3K/AKT signaling. The parathyroid hormone type 1 receptor (PTHR1), the main receptor of PTH, was found to be the upstream of PI3K/AKT signaling, through interfering PTHR1 expression with a small interference RNA following detection of the PI3K/AKT activation. Conclusion Collectively, our study demonstrated that MY-1 accelerates skin wound re-epithelialization by inducing keratinocyte migration and EMT via PTHR1-PI3K/AKT axis activation.

Neuro-Inspired Biomimetic Microreactor for Sensory Recovery and Hair Follicle Neogenesis under Skin Burns

“…To verify whether the protein loaded in Neuro-MOF induces EMT, we selected human immortalized keratinocytes (HaCaT cells) treated with Neuro-MOF to test their EMT, proliferation, and migration capabilities. It is a commonly used cellular model for examining the EMT of keratinocytes during skin wound healing. − As shown in Figure d–f, fibronectin and Vimentin, as markers of EMT, are significantly upregulated under the action of Neuro-MOF, whether in fluorescent confocal observation or Western blotting. It confirms the effect of Neuro-MOF on the induction of EMT in HaCaT cells.…”

Human Parathyroid Hormone (1–34) accelerates skin wound healing through inducing cell migration via up-regulating the expression of Rac1

“…Additionally, it also exerts effects on angiogenesis and extracellular matrix (ECM) deposition [ 4 , 12 , 13 ]. However, the definitive role of hPTH(1–34) in skin-wound healing and the underlying mechanisms remain unclear, although a few studies reported that PTH or PTH analogs (PTHrP2 and MY-1) led to a quicker wound closure [ 4 , 14 – 16 ]. To date, although the in-vivo application of PTH has been under-reported, there is a paucity of detailed experiments that demonstrate the definitive effects of PTH on fibroblasts and keratinocytes.…”

Correction: Human Parathyroid Hormone Analog (3–34/29–34) promotes wound re-epithelialization through inducing keratinocyte migration and epithelial–mesenchymal transition via PTHR1-PI3K/AKT activation

“…Following publication of the original article [ 1 ], the authors wish to clarify their affiliations. The affilaitions in this correction article are displayed correctly (specifically affiliation 1 and 3).…”