In Vitro and In Vivo Evaluation of the Biocompatibility and Cytotoxicity of Local Hemostatic Agents

Abstract

When local hemostatic agents are used in surgery, rapid dissolution followed by prompt absorption without adverse effect after successful hemostasis are essential qualities. Residual hemostatic materials greatly influence host cells during the wound healing process. Biocompatibility of material is also essential. Furthermore, hemostatic agents also should be free of cytotoxicity that may block mitosis and migration of host cells, so that wound healing can proceed smoothly. For the evaluation of biocompatibility and cytotoxicity, 4 commercially available hemostatic agents; oxidized regenerated cellulose (Surgicel(R)) , gelatin sponge (Spongel(R)), microfibrillar collagen (Avitene(R)) and cotton type collagen (Integran(R)) were tested in vitro and in vivo. The hydrogen ion concentration (pH) of culture medium containing hemostatic agents was measured. Fibroblasts were cultured with the hemostatic agents in petri dishes for 5 days. A rabbit ear chamber (REC) model was used to evaluate tissue compatibility and the healing process. Each hemostatic agent was placed in the REC and evaluated macroscopically once a week up to 5 weeks. At 72h, the pH of the culture medium containing Surgicel was low at 7.2, while they stayed between 7.7-7.8 with the other agents. In the fibroblast culture containing Surgicel, cell detachment occurred and the cell numbers decreased, while no particular changes occurred with other hemostatic agents. In the REC model, after 5 weeks Surgicel was dissolved and remained in the effusion, and the healing process was disturbed by inflammation. Spongel was dissolved and absorbed, with normal vasculature. Avitene was dissolved and remained in the effusion, but did not induce strong inflammation. With Integran, the healing process was prompt but the material was still recognizable at 5 weeks. The 4 hemostatic materials tested showed differences in biocompatibility and cytotoxicity. The ability of hemostasis is important; however, after hemostasis is achieved, unused hemostatic material should be eliminated, leaving as little hemostatic agent as possible to avoid postoperative complications.

Evaluation of cytotoxicity of a new hemostatic agent Ankaferd Blood Stopper® using different assays

“…34 The ideal hemostatic agents also should be free of cytotoxicity, but the ability of hemostasis is more important; however, after hemostasis is achieved, unused hemostatic material should be eliminated, leaving as little hemostatic agent as possible in order to avoid postoperative complications. 35 Odabas et al studied that ABS was cytotoxic to human pulp fibroblasts by MTT (3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide, a yellow tetrazole) assay. The influence of cytotoxicity to human pulp fibroblasts depended on the concentration of ABS.…”

“…The ideal hemostatic agents also should be free of cytotoxicity, but the ability of hemostasis is more important; however, after hemostasis is achieved, unused hemostatic material should be eliminated, leaving as little hemostatic agent as possible in order to avoid postoperative complications. 35…”

Cytotoxicity of a new hemostatic agent on human pulp fibroblasts in vitro

“…They reported that the mean clinical and radiographic success rates of pulpotomy treatment with ferric sulphate were 91.6 % and 73.5 %, respectively. The ideal hemostatic agents also should be free of cytotoxicity, but the ability of hemostasis is more important; however, after hemostasis is achieved, unused hemostatic material should be eliminated, leaving as little hemostatic agent as possible in order to avoid postoperative complications (29). Moreover, it should be discussed into a broader perspective that many materials such as peroxides for dental bleaching and bonding agents used in adhesive dentistry all demonstrate cytotoxicity in vitro (30), yet they form an important part of every dentist's restorative armamentarium.…”

Use of an absorbable haemostatic gauze product to control bleeding in 20 horses

“…Acidic products may provoke a strong inflammatory response at the implanted site and slow down the healing process (Ibarrola et al 1985; Krishnan et al 2004; Meric et al 2016). In addition, studies have shown that other ORC products, made from different materials than the studied product, are not entirely biodegradable for as long as 5 weeks after implantation (Tamargo et al 1989; Alpaslan et al 1997; Ibrahim et al 2002; Tomizawa 2004).…”