Experimental and clinical applications of fibrin glue

A 2-year experience with laboratory and clinical applications of fibrin glue is presented. An autologous technique, which eliminates the danger of multidonor preparations, has been developed in our blood bank. While one can obtain different fibrinogen concentrations from the same amount of a patient's blood, in vitro mechanical testing demonstrated that at higher fibrinogen concentrations there is an increase in shear adhesive strength. Evaluation of skin-graft take in 16 Sprague-Dawley rats did not demonstrate significant differences in healing when adhesive use was compared with suture technique. In a clinical study, four different groups of patients (facial burns, hand burns, difficult graft sites, and miscellaneous surgical applications) benefited from autologous or single-donor fibrin glue for a total of 82 cases. There are several distinct advantages to the use of fibrin adhesive: The autologous technique eliminates the risk of transmissible viral diseases (AIDS, hepatitis); it can be used as a sealant in the treatment of seromas, dural leaks, and lymphoceles; and it improves hemostasis and early graft adherence. Face and hands are resurfaced with sheet grafts in a single procedure, obtaining a better aesthetic result with complete graft take and immediate start of physical therapy. Neither sutures nor pressure dressings are required. The minimal postoperative care associated with early return to normal activities seems to increase the satisfaction of patients and nurse personnel.     

The use of aerosolized fibrin glue in face-lift surgery

The purpose of this study was to report the use of aerosolized fibrin glue in face-lift surgery. A prospective study was conducted of 48 patients undergoing face-lift surgery sequentially assigned into two groups. The first 24 patients underwent face lifts without glue and the next 24 patients with the use of aerosolized fibrin glue. One surgeon (J.P.F.) performed all the face lifts using the same technique. Drains were only used in those patients who did not receive fibrin glue. The amount of bruising and edema was compared in the two groups, as was the incidence of complications, such as hematomas. Operating time was also assessed in the two groups. The patients in whom glue was used had significantly less bruising and swelling (p < 0.0001), with a more rapid healing response. The risk of hematoma was also less with the use of glue (0 percent) than without glue (8.3 percent), but this was not statistically significant (p = 0.489). Another benefit was that drains were not needed when glue was used. Operating times were shorter by 13.3 minutes with the use of glue (p < 0.0001). Aerosolized fibrin glue has great promise in improving face-lift results, with excellent outcomes and fewer complications. The added cost of the glue is partially offset by an expedited patient recovery period without the need for drains.     

Experimental studies in animals on the use of a fibrin glue from the human plasma fraction Cohn I in nerve reconstruction

In order to compare the effects of suture and glue direct nerve connections and nerve transplantations in the sciatic nerve in rats were performed. 4, 6 and 12 weeks later, the nerve anastomosis were histologically studied. In the direct nerve connections, despite a single holding suture, dehiscences were frequently detected with the penetration of the adhesive between the nerve ends. Due to the exact adaptation with the aid of inserted nerve grafts, the anastomoses could be repaired sutureless successfully using a fibrin glue cuff. Since histologically, in comparison to suture, no foreign body granulomas were found, the findings in the literature could be confirmed. No cicatricial contractions of the anastomoses could be found. In the interfascicular nerve transplantation, comparatively good results may be obtained using the two-component adhesive on the Cohn I-fraction basis.     

The effect of fibrin on cultured vascular endothelial cells

The normal cobblestone monolayer architecture of cultured vascular endothelium becomes rapidly disorganized after contact of the cell layer with a fibrin clot. The cells of a confluent endothelial monolayer separate into individual migratory cells in 4–6 hr after contact with fibrin. The effect is reversible in that removal of the fibrin clot results in resumption of the normal morphology within about 2 hr. No other cell type tested exhibits the same change in organization when exposed to fibrin. A similar morphological change in endothelium does occur after the cell layer is overlaid with a collagen fibril gel but a gel of methylcellulose has no effect. It is proposed that the change in behavior of endothelial cells in response to contact with fibrin may represent a cellular component of fibrinolysis. The implications of this finding for the pathophysiology of disease states involving intravascular fibrin deposition are discussed.     

On the use of phase partition to isolate vacuoles from cultured cells

Vacuoles isolated from cultured rose-cell protoplasts by controlled lysis were subjected to partition in polyethylene glycol-dextran two-phase systems. The vacuoles showed a strong affinity for the polyethylene glycol phase, like plasma membrane and unlike other membraneous organelles. After phase partition at 4°C, dilution of the polyethylene glycol destroyed a large proportion of the vacuoles. After phase partition at room temperature (ca. 20°C), the vacuoles in the polyethylene glycol phase were relatively stable to dilution. Two-phase partition may be useful for purification of intact vacuoles and possibly of tonoplast membranes. Minor proportions of acid phosphatase and -mannosidase were associated with these vacuoles.     

The use of haptenated-immunoglobulin molecules to induce tolerance in B cells from neonatal mice

The role of the Fc region of trinitrophenylated (TNP)-immunoglobulins (Ig) in their ability to induce tolerance in immature B cells was examined. With the use of B cells from neonatal mice, tolerogens that could or could not bind to Fc receptors were assessed for their ability to induce tolerance. This was accomplished by tolerizing spleen cells in bulk culture and assessing the degree of tolerance by challenging the cells with the thymus-independent antigen TNP-Brucella abortus (TNP-BA) in limiting dilution cultures. It was found that by using tolerogens containing 10 to 11 haptens per Ig molecule, immature B cells were very susceptible to tolerance induction. Mature B cells were not as susceptible. This increased susceptibility was independent of the Fc portion of the tolerogen, because TNP11-HGG and a TNP10-F(ab')2 induced equivalent degrees of unresponsiveness. When the TNP density was lowered to approximately five haptens per Ig molecule, those Ig molecules that contained Fc portions were superior tolerogens with the use of B cells from 6-day-old mice. Thus, a TNP4-HGG, TNP7-mouse IgG1, and TNP6-mouse IgG2a were more effective tolerogens than either TNP5-F(ab')2 or TNP6-mouse IgG3. These results confirm previous findings that immature B cells are inherently more susceptible to tolerance induction than mature B cells. They also suggest that very lightly haptenated Ig molecules may depend on Fc receptor-binding for effective tolerance induction. Finally, by means of a cytofluorograph, the surface IgD (sIgD) and sIgM phenotypes of splenic B cells from neonates of increasing age were determined. When comparing the phenotype of maturing cells with their tolerance susceptibilities, a correlation between the appearance of sIgD and the acquisition of resistance to tolerance was observed.     

Liposomes induce chromosome aberrations in human cultured cells

The genotoxic effect of multilamellar lipid vesicles (MLV) was analysed on cultured heteroploid and diploid human cells. Dose-dependent reduction of cell survival and mitotic rate as well as induction of chromosome aberrations were observed. Chromatid and chromosome breaks and chromatid exchanges were found in 24-h culture after liposome treatment, whereas chromosome rearrangements were prevalent at 48 h. Neutral (PC/Chol) and positive (PC/SA) MLV showed a greater damage than negative (PS/PC; PS) MLV. Fibroblasts were the most sensitive cell type. In the case of PC/Chol MLV vesicles, control experiments with PC and Chol of controlled purity ruled out the possibility that the observed chromosome aberrations were caused by toxic oxidation products present in commercial preparations.     

Use of fibrin glue in sucrose gap method

Isolated frog nerves or rabbit sinus node strips were mounted in a single sucrose gap chamber. A fibrin glue Tissucol was used in this arrangement as an intercompartment seal. Under such conditions, the specimens produced stable trans-gap action potentials of relatively high amplitude. In other experiments the gap was filled with fibrin in place of sucrose solution. The results obtained in the fibrin gap experiments resembled these mentioned above.     

PDGF-BB and IGF-I use different signaling pathways to induce fibronectin synthesis in cultured rat thoracic aortic smooth muscle cells

Fibronectin, an extracellular matrix protein, acts as an early signal in initiating cell proliferation. Results have indicated that platelet-derived growth factor BB (PDGF-BB) and insulin-like growth factor-I (IGF-I) both enhance fibronectin gene expression. Genistein inhibits PDGF-BB-induced fibronectin levels without inhibiting IGF-I-induced fibronectin levels. It indicates that PDGF-BB and IGF-I utilize separate signaling pathways to induce the synthesis of fibronectin.     

PDGF-BB and IGF-I use different signaling pathways to induce NaK-ATPase subunits in cultured rat thoracic aortic smooth muscle cells

(Na(+)+K(+))-adenosine triphosphatase (NaK-ATPase), an ubiquitous membrane transport protein consisting of alpha and beta subunits, regulates Na(+)/K(+)fluxes and maintains many vital physiological functions, including cell growth. Results have indicated that platelet-derived growth factor (PDGF) and insulin-like growth factor-I (IGF-I) both enhance NaK-ATPase subunits. Genistein, an inhibitor of tyrosine phosphorylation, inhibits serum- and PDGF-BB-induced NaK-ATPase alpha(1)subunit protein levels without inhibiting IGF-I-induced NaK-ATPase alpha(1)subunit protein levels. These results indicate that PDGF-BB and IGF-I utilize separate signaling pathways to induce the synthesis of NaK-ATPase alpha(1)subunits. In addition, genistein failed to inhibit PDGF-BB-stimulated NaK-ATPase beta(1)subunit levels, suggesting that two separate pathways are involved to induce the synthesis of the NaK-ATPase alpha(1)and beta(1)subunits, respectively.     

The use of fibrin glue in skin grafts and tissue-engineered skin replacements: a review.

Fibrin glue has been widely used as an adhesive in plastic and reconstructive surgery. This article reviews the advantages and disadvantages of its use with skin grafts and tissue-engineered skin substitutes. Fibrin glue has been shown to improve the percentage of skin graft take, especially when associated with difficult grafting sites or sites associated with unavoidable movement. Evidence also suggests improved hemostasis and a protective effect resulting in reduced bacterial infection. Fibrin, associated with fibronectin, has been shown to support keratinocyte and fibroblast growth both in vitro and in vivo, and may enhance cellular motility in the wound. When used as a delivery system for cultured keratinocytes and fibroblasts, fibrin glue may provide similar advantages to those proven with conventional skin grafts. Fibrin glue has also been shown to be a suitable delivery vehicle for exogenous growth factors that may in the future be used to accelerate wound healing.