Chondrocyte suspension in fibrin glue

Autologous chondrocyte implantation has been shown to be a promising method for treatment of deep articular cartilage defects. The hyaline cartilage formed by implanted autologous chondrocytes has biomechanical properties similar to those of natural articular cartilage. Between June 2006 and September 2008 we performed Autologous chondrocyte implantation (ACI) in 50 patients and the chondrocytes were supported in fibrin glue. The cartilage biopsy samples were taken from the non-weight bearing area of the patient’s femoral condyle and the samples were transferred to the cell culture laboratory. Chondrocyte were kept in culture about 20 days. Fibrin glue was used as a three dimensional carrier for chondrocyte implantation. A 450 ml of patient’s own blood was collected prior to transplantation to produce autologous fibrinogen. Alternatively the allogenic fibrinogen was prepared from Regional Blood Center voluntary donors. Before surgery the chondrocyte suspension was mixed with fibrin glue and gel—like fibrograft was prepared. The total number of cells and the size of fibrograft depended on the defect size in the knee. Our results suggest that ACI technique with fibrin glue is a promising method for treatment of cartilage defect.     

Experimental use of fibrin glue to induce site-directed osteogenesis from cultured periosteal cells

The purpose of this study was to determine whether a combination of fibrin glue and cultured periosteal cells will result in new bone formation at heterotopic sites in nude mice. Growing cells and developing matrices surrounding periosteal explants from the diaphyses of radii of newborn calves were minced and mixed with fibrin glue in a syringe. The cell/matrix-fibrin glue admixture was then injected into the subcutaneous space on the dorsum of athymic nude mice. After 12 weeks of implantation, gross morphology and histologic investigations showed newly formed bone structures in all cell/matrix-fibrin glue admixtures, but none in fibrin glue injected alone and used as control samples. Osteopontin, a protein important in bone development, was identified by a Western blot assay of the cell/matrix-fibrin glue composite. This study supports the feasibility of initiating site-directed formation of bone structures at heterotopic tissue sites by means of injection of cultured periosteal cells and matrix in a fibrin glue carrier.     

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.     

Salmon fibrin glue in rats: Antibody studies

Fibrin sealants and topical thrombin preparations are often used for haemostatic and sealing applications in clinical practice. Some of these preparations contain coagulation factors from bovine sources. To minimize the risk of infection and immunogenicity connected with mammalian blood products, proteins derived from the plasma of farmed Atlantic salmon have been considered as an alternative to these mammalian sources. The purpose of this study is to characterize the immunogenicity of salmon fibrin glue in an animal model focusing on crossreactivity of IgG antibodies to host endogenous counterparts. After two immunizations with salmon fibrin glue, rats developed antibodies of IgG and IgM type to both fibrin glue components. Weak crossreactivity to endogenous fibrinogen and thrombin was seen in a subset of rats after the second application of salmon proteins. Coagulation tests showed that salmon fibrin application has no effect on coagulation profiles in mammalian hosts, consistent with previous reports that found no evidence of significant crossreactivity with host proteins. These studies support the potential suitability of salmon fibrin glue for the development of preparations with clinical impact. Before human use can be considered, however, additional data about safety of this preparation in other animal models, including large animal studies, should be obtained.     

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.     

Evaluation of fibrin glue in rat sciatic nerve repairs

Using the rat sciatic nerve model, we evaluated the merits of homologous fibrin glue in the repair of peripheral nerve transections as compared to standard epineural suture repairs. A total of four study groups were used, with 10 animals assigned to each group. In group I, the transected sciatic nerve was repaired with six interrupted 10-0 nylon sutures; in group II, only two interrupted sutures were used; in group III, a two-suture repair was reinforced with fibrin glue; and in group IV, only fibrin glue was used. All animals were sacrificed at 8 weeks, and histologic sections evaluated. When fibrin alone was used, dehiscence occurred in 80 percent of the animals, and as reinforcement of a two-suture repair, it only increased the inflammatory reaction.     

Injectable tissue-engineered cartilage using a fibrin glue polymer.

The purpose of this study was to demonstrate the feasibility of using a fibrin glue polymer to produce injectable tissue-engineered cartilage and to determine the optimal fibrinogen and chondrocyte concentrations required to produce solid, homogeneous cartilage. The most favorable fibrinogen concentration was determined by measuring the rate of degradation of fibrin glue using varying concentrations of purified porcine fibrinogen. The fibrinogen was mixed with thrombin (50 U/cc in 40 mM calcium chloride) to produce fibrin glue. Swine chondrocytes were then suspended in the fibrinogen before the addition of thrombin. The chondrocyte/polymer constructs were injected into the subcutaneous tissue of nude mice using chondrocyte concentrations of 10, 25, and 40 million chondrocytes/cc of polymer (0.4-cc injections). At 6 and 12 weeks, the neocartilage was harvested and analyzed by histology, mass, glycosaminoglycan content, DNA content, and collagen type II content. Control groups consisted of nude mice injected with fibrin glue alone (without chondrocytes) and a separate group injected with chondrocytes suspended in saline only (40 million cells/cc in saline; 0.4-cc injections). The fibrinogen concentration with the most favorable rate of degradation was 80 mg/cc. Histologic analysis of the neocartilage showed solid, homogeneous cartilage when using 40 million chondrocytes/cc, both at 6 and 12 weeks. The 10 and 25 million chondrocytes/cc samples showed areas of cartilage separated by areas of remnant fibrin glue. The mass of the samples ranged from 0.07 to 0.12 g at 6 weeks and decreased only slightly by week 12. The glycosaminoglycan content ranged from 2.3 to 9.4 percent for all samples; normal cartilage controls had a content of 7.0 percent. DNA content ranged from 0.63 to 1.4 percent for all samples, with normal pig cartilage having a mean DNA content of 0.285 percent. The samples of fibrin glue alone produced no cartilage, and the chondrocytes alone produced neocartilage samples with a significantly smaller mass (0.47 g at 6 weeks and 0.46 g at 12 weeks) when compared with all samples produced from chondrocytes suspended in fibrin glue (p < 0.03). Gel electrophoreses demonstrated the presence of type II collagen in all sample groups. This study demonstrates that fibrin glue is a suitable polymer for the formation of injectable tissue-engineered cartilage in the nude mouse model. Forty million chondrocytes per cc yielded the best quality cartilage at 6 and 12 weeks when analyzed by histology and content of DNA, glycosaminoglycan, and type II collagen.     

The effect of fibrin glue on skin grafts in infected sites.

Fibrin bonding of skin grafts to wounds is an essential part of the graft-adherence process. Bacteria, in concentrations greater than 10(5)/gm of tissue, are associated with graft failure. Sixty-five rats were randomly divided into three groups, dorsal split-thickness skin grafts were harvested, and the sites were inoculated with Staphylococcus aureus. After incubation, each wound was quantitatively biopsied and treated with saline, fibrin glue with aprotinin, or fibrin glue alone. We found that the addition of commercially available fibrin glue with or without the antifibrinolytic agent aprotinin is capable of restoring graft adherence to normal levels in graft sites infected with greater than 10(5) bacteria/gm of tissue. Fibrin glue may have potential for increasing skin-graft take in the clinical situation where the graft bed is infected.     

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.