Vitamin E triggers poly(2-hydroxyethyl methacrylate) (PHEMA) embolic potential: a proposed application for endovascular surgery.

Poly(2-hydroxyethyl methacrylate) (PHEMA) is a biocompatible polymer used as embolizing agent for endovascular surgery. Blending of PHEMA with a hydrophobic and anti-oxidant agent, Vitamin E (Vit.E, 0.1-10%, w/v), modified PHEMA's haemocombatibility, evaluated measuring wettability, plasma protein adsorption along with whole blood coagulation time. The presence of Vit.E increases PHEMA's hydrophobicity and plasma protein adsorption (in particular albumin and Immunoglobulin G), while it also accelerates blood clot formation. These effects are developed due to a combination of issues such as surface hydrophobicity and plasma protein adsorption induced by the presence of Vit.E, suggesting that Vit.E blending could improve the use of PHEMA as embolizing agent.

Mechanical stretching modulates growth direction and MMP-9 release in human keratinocyte monolayer.

Cells within human skin are exposed to mechanical stretching that is considered a trigger stimulus for keratinocyte proliferation, while its effect on keratinocyte migration has been poorly investigate. In order to explore the effect of stretching on keratinocyte migration spontaneously immortalized human keratinocyte (HaCaT) monolayers seeded onto collagen I-coated silicon sheets were stimulated 3 times for 1 hour every 24 hours (total time = 72 hours) by mechanical stretching increasing substrate deformations (10%) applied both as static (0 Hz) and cyclic (0.17 Hz) uniaxial stretching. At the end of stimulations monolayer areas measured in both static and cyclic samples appeared reduced and strongly oriented in a direction perpendicular to the stress direction compared to unstimulated ones. Moreover during the mechanical stimulation period HaCaT monolayers strongly increased the release in the medium of matrix metalloproteinase 9 (MMP-9), a proteolytic enzyme necessary for keratinocyte migration.

Adsorption of matrix metalloproteinases onto biomedical polymers: a new aspect in biological acceptance.

Matrix metalloproteinases (MMPs) are zinc-dependent enzymes involved in the remodelling of connective tissues during the development and wound healing. Moreover, two MMPs, Gelatinase A (MMP-2) and Gelatinase B (MMP-9), are also present in body fluids such as blood and urine and, therefore, they can be in contact with implanted biomaterials and can be adsorbed onto their surface. In order to test this hypothesis disks of different polymers (polystyrene (PS), polyvinyl chloride (PVC), poly(D,L-lactide) (PLA), polymethyl methacrylate (PMMA) and poly(2-hydroxyethyl methacrylate) (PHEMA)) have been exposed to human plasma and adsorbed proteins have been eluted and analyzed. Using Western blot and substrate zymography analysis, we observed that both MMP-2 and MMP-9 adsorbed onto the surfaces of all the polymers, especially hydrophilic ones (PMMA and PHEMA) and PLA, in both the active and inactive forms. Furthermore, we observed that adhesion of human granulocyte neutophils to PMMA, the polymer that adsorbed the higher quantity of MMP-2 and MMP-9 compared to the others, was reduced by more that 50% by the presence of a gelatinase inhibitor. This data suggest a surprising role of these absorbed enzymes in the adhesion of neutrophil onto some polymeric biomaterials surface and, therefore, in the setting of inflammation.

Haemocompatibility of vitamin-E-enriched poly(D,L-lactic acid).

Poly(D,L-lactic acid) (P(D,L)LA) is a biocompatible and biodegradable polymer whose use is limited to orthopaedic applications. In fact, the mechanical properties of P(D,L)LA are not usually utilized for cardiovascular applications, as the polymer has been proven to activate both granulocyte- and platelet-causing inflammation. In order to improve P(D,L)LA haemocompatibility vitamin E (alpha-tocoferol, 10-30% (w/w)), a natural biological anti-oxidant and anti-inflammatory agent, was added during the solvent casting of P(D,L)LA film. The P(D,L)LA films obtained were then analysed using FT-IR analysis to assess vitamin E presence; polymer surface wettability and human plasma protein adsorption were measured by sessile drop test, spectrophotometric protein quantification and Western blot, respectively, and polymer haemocompatibility was assessed measuring platelet and granulocyte adhesion and whole blood coagulation. Vitamin E presence caused an increase in polymer surface wettability and human plasma protein adsorption. The combination of both effects may account for the decrease in platelet and granulocyte adhesion and for the doubling of whole blood clotting time measured onto vitamin-E-enriched P(D,L)LA compared to control P(D,L)LA. Our results indicate that vitamin E addition improves P(D,L)LA haemocompatibility, making this polymer suitable for cardiovascular application.

Cellular behavior of neointima-like cells onto vitamin E-enriched poly(D,L)lactic acid.

In-stent restenosis is a process that occurs in 10-50% of cases currently treated with stent and it is caused by an abnormal smooth muscle cell (SMC) proliferation and migration in the vascular lumen. One of the most promising strategy to reduce restenosis is stent coating with biodegradable polymers to deliver in situ anti-proliferative drugs. Poly(D,L)lactic acid (P(D,L)LA), one of the most interesting candidate for stent coating, has been observed to induce inflammation and neointimal proliferation. In our laboratory, we developed P(D,L)LA enriched with Vitamin E (Vit.E), an anti-oxidative and anti-inflammatory agent that reduces also SMC proliferation. In order to evaluate the in vitro cellular behaviour of neointima cells onto Vitamin E-enriched P(D,L)LA, cell adhesion and proliferation along with the expression of two SMC migration markers (MMP-9 and hyaluronic acid receptor CD44) were measured in rat vascular SMC A10 cells seeded onto control P(D,L)LA (PLA) and P(D,L)LA films containing 10-30% (w/w) Vit.E (PLA10-30). Cell adhesion, proliferation and MMP-9 production were unaffected by the Vit.E presence in the PLA films after 24 h, while proliferation was slowed or blocked after 48 and 72 h onto PLA10, 20 and 30. MMP-9 production was almost blocked and CD44 density decreased significantly after 72 h for cells grew onto PLA30 compare to cells seeded onto PLA. These data indicate that Vit.E-enriched P(D,L)LA could be an interesting polymer for stent coating.