| Popis: |
Faecal incontinence is a major public health issue that has yet to be adequately addressed. The reported prevalence of postpartum faecal incontinence ranges from 3% to 29%, with obstetric trauma being the most common cause of faecal incontinence. Trauma is often attributed to injury of the anal sphincter muscles. New therapies to restore continence are emerging which combine minimally invasive procedures with restoration of functional sphincter muscle. The aim of the research undertaken in this thesis was to evaluate the use of poly(D,L-lactide-co-glycolide) thermally induced phase separation (TIPS) microspheres for delivery of muscle precursor cells to regenerate injured skeletal muscle. The objectives were as follows: (a) to test the feasibility of attaching muscle precursor cells to the surface of TIPS microspheres; (b) to characterize the phenotypic status of cells attached to the TIPS microspheres; (c) to demonstrate targeted cell delivery using TIPS microspheres; and (d) to investigate the engraftment of cells delivered via TIPS microspheres into host tissue. Major findings of this thesis indicated that the development of a static–dynamic culture technique successfully attached muscle precursor cells to the surface of TIPS microspheres. The technique produced a uniform distribution of cells across the surface of the TIPS microsphere within 24 hours. Cells attached to TIPS microspheres were phenotypically characterized at a gene and protein level. Muscle precursor cells were demonstrated to show an increase in both mRNA and protein expression of cell contractile apparatus. The delivery technique showed no apparent cell membrane damage upon delivery, with substantial cell migration off the TIPS microspheres for delivery in vitro and in vivo. Preliminary findings of an in vivo delivery study showed localized delivery, migration, integration of cells with host tissue, and evidence of myotube formation. The current research supports further study of muscle precursor cells delivered via TIPS microspheres. The concepts developed in this thesis offer a novel approach to regenerating a skeletal muscle tear for obstetric trauma-induced faecal incontinence. |
