Platelet-Rich Plasma Promotes the Proliferation of Human Keratinocytes via a Progression of the Cell Cycle. A Role of Prolidase

Abstract

Although the role of platelet-rich plasma (PRP) in tissue regeneration has been confirmed in many studies, the mechanism of this process is still not fully understood. Human keratinocytes (HaCaT) cells were used as an experimental model for studies on the effects of PRP on cell proliferation, migration, collagen biosynthesis, prolidase activity, and its expression and anabolic signaling. The activation of epidermal growth factor receptor (EGFR), β1-integrin, and insulin-like growth factor-1 receptor (IGF-1R) by PRP were investigated by western blot and immunocytochemistry. It has been found that PRP induced keratinocytes migration and proliferation through activation of cell cycle progression and EGFR downstream signaling. Similar biological effects were achieved by an addition to the culture medium of prolidase (PEPD), a ligand of EGFR (PRP is a rich source of PEPD–2 ng/mL). PRP-dependent stimulation of collagen biosynthesis was accompanied by an increase in the expression of NF-κβ, IGF-1R-downstream signaling proteins, and PEPD activity. The data suggest that PRP activates a complex of growth factors and adhesion receptors that stimulate cell proliferation, migration, and collagen biosynthesis. PRP induces PEPD-dependent human keratinocyte proliferation through activation of the EGFR receptor. Our study provides a novel mechanism of PRP-dependent wound healing.

Recombinant Prolidase Activates EGFR-Dependent Cell Growth in an Experimental Model of Inflammation in HaCaT Keratinocytes. Implication for Wound Healing

“…Under chemically-induced cell disruption PEPD concentration significantly increases ( Yang et al, 2013 ). Another possible source of prolidase can be platelets in the bloodstream ( Guszczyn et al, 2017 ; Misiura et al, 2021a ). Platelets are essential players in the initial stage of inflammation as they carry various inflammatory mediators.…”

PRODH/POX-Dependent Celecoxib-Induced Apoptosis in MCF-7 Breast Cancer

“…Cells were cultured in FBS-free DMEM with Cx (5–25 µM) for 24 h. The procedure for harvesting the cells was performed as previously described [ 31 ]. The supernatant was aliquoted and stored at −80 °C.…”

“…Western blot analysis was carried out as described by Misiura et al [ 31 ]. The membranes were incubated with primary antibodies diluted 1000 times in 5% bovine serum albumin (Sigma Aldrich, Saint Louis, MO, USA) in TBS-T (20 mM Tris, 150 mM NaCl, 0.1% Tween-20, pH 7.6).…”

“…Cells were cultured in FBS-free DMEM with Cx (5-25 µM) for 24 h. The procedure for harvesting the cells was performed as previously described [31]. The supernatant was aliquoted and stored at −80 • C. Protein concentration was measured using the Pierce BCA assay kit (Thermo Fisher Scientific, Waltham, MA, USA).…”

Proline Dehydrogenase/Proline Oxidase (PRODH/POX) Is Involved in the Mechanism of Metformin-Induced Apoptosis in C32 Melanoma Cell Line

“…The cell cycle of C32 WT and C32 POX− cells upon metformin treatment (0-20 mM) for 24 h was analyzed as previously described [69].…”