Critical Limb Ischemia (CLI) / Peripheral Arterial Occlusive Disease (PAOD)
Conditions
Interventions
None listed
Sponsors
Universitair Medisch Centrum Utrecht
Eligibility
Age
18 Years to 99 Years
Inclusion criteria
Inclusion criteria: - Age >18 years; - Patients undergoing surgical procedures during which bony structures containing active bone marrow are encountered (eg pelvic girdle, caput femoris, acetabulum, distal femur or proximal tibia); - Scheduled for surgical intervention; - Approval of both the anaesthetist and the surgeon performing the surgical procedure; - Written informed consent.
Exclusion criteria
Exclusion criteria: - History of overt cardiovascular disease; - Major trauma involving multiple bones or damaged internal organs; - Known disease originating from the bone marrow (ie leukaemia, lymphoma, metastatic disease); - Chronic autoimmune disease (ie SLE, rheumatic arthritis etc); - Known infection with HIV, hepatitis B or C virus.
Design outcomes
Primary
| Measure | Time frame |
|---|---|
| The main study parameters tested in this study are the progenitor cell levels, of EPC in particular, in BM and blood of healthy controls and how they relate to levels in patients with CLI. Cells will be counted and characterized using fluorescence activated cell sorting (FACS). This will be performed using the same protocols as for patients suffering from CLI, which participate in the Juventas trial. With this method the amount of EPCs can be assessed as well as certain other cell populations potentially involved in vascular health and neovascularization. Antibodies to CD34, CD31, KDR, CD133, CD45, CD184, CD140b, CD14 and CD26 will be used. | — |
Secondary
| Measure | Time frame |
|---|---|
| Secondary parameters in this study apply to mechanisms involved in EPC mobilization and function, such as BM and PB levels of growth factors (VEGF, EGF, bFGF, SDF-1a etc), cytokines (IL-6, IL-2 etc) and other mediators such as proteases (MMP*s) and nitric oxide related mechanisms (eNOS, NO, MAPK etc). Additionally, specific cell populations will be isolated (ie mononuclear cell fraction [MNC]) and cultured according to standardized pre-tested culture protocols to obtain mesenchymal stem cells (MSCs; from BM-MNC) and circulating angiogenic cells (CACs; obtained from PB-MNC). The former being involved in progenitor cell mobilization and angiogenic processes and the latter deemed to be important for paracrine stimulation of angiogenesis. Different BM derived subfractions (eg BM-MNCs, CD14+, CD34+, and MSCs) will be studied for there in vitro migratory function, using trans-well assays, and in vivo neovascularization stimulating potential in an animal model (hindlimb ischemia mouse model). Studying levels and function of these cells from healthy donors compared to patients with CLI will provide detailed insights in whether and how the function of (endothelial) progenitor cells is impaired in patients with CLI. This improved knowledge on disturbed mechanisms will be a foundation to elucidate how we can further improve cell-based therapies in patients with cardiovascular diseases, CLI in particular. These quantitive and functional assays will enable an as optimal as possible use of the materials obtained in the Juventas trial, as well. | — |
Countries
Netherlands
Outcome results
None listed