Induced Regenerative Elastic Matrix Repair in LOXL1 Knockout Mouse Cell Cultures: Towards Potential Therapy for Pelvic Organ Prolapse

Abstract

Impaired elastic matrix remodeling occurs in reproductive tissues after vaginal delivery. This has been linked to development of pelvic organ prolapse (POP) for which there currently is no pharmacologic therapy. We have previously shown that including hyaluronan oligomers and transforming growth factor beta 1 (termed elastogenic factors, EFs), significantly enhance tropoelastin synthesis, elastic fiber assembly, and crosslinking by adult vascular smooth muscle cells (SMCs). The goal of this study was to ascertain if these factors similarly improve the quantity and quality of elastic matrix deposition by vaginal SMCs (VSMCs) isolated from lysyl oxidase like-1 knock out (LOXL1 KO) mouse model of POP. Cells isolated from whole vagina of a LOXL1 KO mouse (multiparous, stage 3 prolapse) were cultured and identified as SMCs by their expression of various SMC markers. Passage 2 vaginal SMCs (VSMCs; 3×10 4 /10 cm 2) were cultured for 21 days with EFs. Cell layers and spent medium aliquots were assessed for elastin content and quality. EF treated VSMCs proliferated at a similar rate to untreated controls but synthesized more total elastin primarily in the form of soluble matrix elastin. Elastin mRNA was also increased compared to controls. The elastic matrix was significantly denser in EF-treated cultures, which was composed of more mature, non-interrupted elastic fibers that were absent in controls. The results are promising towards development of a therapy to enhance regenerative elastic matrix repair in post-partum female pelvic floor tissues.