Background. In 2003 we started investigations concerning the possibility of using new techniques, including bone-marrow cell implantation, in the therapy of critical leg ischaemia. Safety procedures, scheme of work and inclusion and exclusion criteria were established during the first faze in collaboration with various local departments and laboratories.
Material and methods. Inclusion criteria consisted of critical leg ischaemia, no possibility of surgical operation, no effective 2 months conservative treatment or the possibility of limb amputation in the following months. Exclusive criteria included causes of ischaemia other than atherosclerosis, absence of consent, need for urgent amputation, cancer or poor general condition. After acquiring conscious consent, patients were randomized into two groups: conventional therapy (control group) (n = 15), and those treated with bone-marrow cells (n = 14). Standard bone marrow collection was performed as early as the first operation (usually about 8 a.m.) under general anaesthesia. Material was then prepared filtration and concentration in a separator - using standard procedure. Morphology and measurement of the amount of nuclear cells and CD34 cells were performed as well as microbiological examination. This material, including progenitor cells, was injected using two methods about 46 hours after bone marrow aspiration. In every case it involved several intramuscular injections into the ischemic limb (thigh and leg). In 4 cases we performed intravascular placement of progenitors using arteriography (in the distal part near the occluded part of the axial limb artery).
Results. Observation was based on several parameters: vital parameters (blood pressure, heart rate, respiratory rate, temperature) measured every day during hospitalisation; ankle/brachial index (ABI), ulcerations or pedal necrosis documented by photography and subjective parameters: feeling of pain (VAS), self care (WHO scale) measured 1, 3 and 6 months after our procedure. Baseline angiography was performed in every case. Control angiography followed bone marrow collection after 3 months in 5 cases. No changes in angiograms were detected. Endpoints were: amputation (10) and death (no observations). In case of amputation, material was histologically examined and signs of angiogenesis (mostly new vein formation) were detected. In the group treated by stem cell implantation we reached greater subjective improvement in 12 cases compared to 7 in the control group; healing of ulcers in 8 cases (complete in 5 cases) compared to 3 cases in the control group. Amputations were performed in 3 patients in the index group compared to 7 in the control group.
Conclusions. We think that implantation of patients' bone-marrow progenitor cells in cases of critical leg ischaemia is safe and is a prospective therapeutic method.