Biomedical subjects

Alberto Balbarini

Publications and source records attributed to Alberto Balbarini.

21 records · Page 2Linked to original sources

PubMedDate not supplied

Different growth conditions for peripheral blood endothelial progenitors.

PURPOSE: To compare different growth conditions for endothelial progenitor cells (EPCs) from peripheral blood mononuclear cells (PBMNCs). METHODS AND MATERIALS: PBMNCs of healthy volunteers were cultured on fibronectin as follows: M199 with VEGF, bFGF, IGF-I; the same medium with bovine retina-derived extract (RDE); freshly isolated or depleted of adherent cells PBMNCs in HUVEC conditioned medium; DiI-stained PBMNCs with HUVECs (1:4 ratio) in Ml99 with RDE. PBMNCs were analysed by FACS using mAbs for endothelial markers. EPCs migration was determined using a modified Boyden chamber assay and VEGF as chemoattractant. EPCs were seeded alone or with HUVECs on Matrigel to assess in vitro angiogenesis. RESULTS: With growth factors, numerous cell clusters appeared within 1 week. Spindle-shaped and attached cells sprouted, differentiating in endothelial cell (EC)-like cells within 2 weeks and forming cobblestone-like monolayers within 3 weeks. With RDE, numerous large cell clusters appeared within 1 week, but the number of cells with an EC morphology decreased during culture. FACS confirmed the endothelial phenotype and attached cells were able to migrate in response to VEGF. When nonadherent cells were cultured in HUVEC conditioned medium, they proliferated readily and EPCs were induced while freshly isolated cells neither proliferated nor induced EPCs. FACS analysis of the cocultures showed the presence of double-labeled PBMNCs expressing endothelial antigens. Capillary-like structures were observed on Matrigel only from cocultures and PBMNCs were able to incorporate in these networks. CONCLUSIONS: PBMNCs are able to differentiate in EPCs when stimulated with appropriate culture conditions (growth factors, HUVEC conditioned medium, HUVECs).

Anticoagulants↗

PubMedDate not supplied

Endothelial progenitor cells induction by short-term stimulation with phytohaemagglutinin of mononuclear cells.

BACKGROUND: The identification of circulating endothelial progenitors cells (EPCs) in the adult has forced to reconsider how new blood vessels grow in physiological and pathological conditions. Neovascularization during adult life has long been attributed to angiogenesis only. However, recent studies have revealed that peripheral blood EPCs may be recruited and incorporated into sites of active neovascularization. PURPOSE: To verify that EPCs are induced from peripheral blood mononuclear cells (PBMCs) and bone marrow derived mononuclear cells (BMMCs) upon short-term stimulation with phytohaemoagglutinin (PHA), a potent T-cell mitogen. METHODS: PBMCs and BMMCs were isolated from healthy donors. Freshly isolated or depleted of adherent cells (one day and three days of adherence) mononuclear cells (MCs) were cultured in RPMI, 10% FBS, containing PHA (10 microl/10(6) cells) for 24 h. After stimulation with PHA, clusters of adherent cells were further propagated in M199 containing L-glutammine, Hepes, 20% FBS, heparin, antibiotics and bovine retina extract for 1 and 2 weeks. PBMCs and BMMCs cultured without PHA stimulation served as controls. FACS of EPCs was performed on attached cells after 7 and 14 days of culture. RESULTS AND CONCLUSION: After stimulation of MCs with PHA for 24 h, many cells clusters were observed and around these clusters some adherent EC-like cells were observed. These cells were ovoid but a very little of these were elongated in morphology, however their number and size gradually increased during culture. However a longer time was needed for obtaining EPCs from MCs harvested after adherence. Thus this indicates that short-term signals provided by PHA must be sufficient for MCs to express the ligands necessary for the induction of EPCs but signals from monocytes/macrophages are important for a more rapid differentiation.

Adult↗

PubMedDate not supplied

Transcutaneous oxygen and carbon dioxide levels with iloprost administration in diabetic critical limb ischemia.

Iloprost, a prostacyclin analogue, is a treatment option for surgically unsuitable diabetic chronic critical limb ischemia (CLI), although its outcome is difficult to be anticipated clinically. Whether transcutaneous (tc) oxygen tension (PO2) predicts the response to iloprost in diabetic CLI is unclear at this point and, in that same context, the prognostic role of tc carbon dioxide tension (PCO2), another ischemia-sensitive parameter, is unknown. Supine and dependent tcPO2 and tcPCO2 were measured at baseline and after 4 weeks of iloprost treatment in 31 limbs of 26 type-2 diabetic angiopathies with CLI not amenable to surgery. Success was defined as pain relief and significant reduction of analgesics. Clinical outcome was stratified by baseline tcPO2 and tcPCO2 tertiles, and likelihood ratios (LR) quantified the increase from pretest chances given a certain result. Iloprost succeeded in 16 (52%) and failed in 15 limbs (48%) and post-treatment tcPO2 followed a parallel course. Failures increased by ascending baseline tcPCO2 and descending tcPO2 tertiles; successes behaved specularly. Predictions of failure based on elevated tcPCO2 (>53 mm Hg) were more efficient than relying on depressed tcPO2 (LR 10.7 vs 3.6); success was almost certain when tcPO2 was >23 mm Hg (LR = 17.8). Dependent determinations were less useful than supine measurements for prognostic use. Elevated tcPCO2 predicted failure efficiently and high tcPO2 was a useful prognostic tool for success of iloprost, suggesting that their combined use may allow better prognostic stratification and improve the therapeutic approach to diabetic CLI.

Aged↗