Red blood cell salvage analysis from clotted blood.

BACKGROUND: Blood clots discovered within body cavities intra-operatively are often manually broken up and placed in an autotransfusion device to recover autologous blood cells. This study evaluated the efficiency at which these red blood cells can be recovered from clot and to determine if these cells would be free of fibrin and clumping which might pose a risk of micro-emboli. MATERIALS AND METHODS: Whole blood was aliquoted into 25 mL volume samples. The blood was then allowed to clot, and after 24 hours the clotted blood was manually kneaded by hand for 1, 2, 3, or 5 minutes. One mL of the harvested blood was fixed and processed for scanning electron microscope imaging. Plasma from the rest of the sample was then separated and underwent spectrophotometry for analysis of relative free haemoglobin. RESULTS: Blood recovered from the clotted blood ranged from 60 to 80% as time increased from 1 to 5 minutes of kneading. Volume of erythrocytes recovered from 1 minute compared to 2 minutes was statistically significant but not significant between 2 minutes or any longer period of time. Imaging did not show any evidence of fibrin strands or significant cell fragmentation. Spectrophotometry showed a steady increase of observed absorption at 540 nm, indicative of free haemoglobin, as manual kneading time increased. DISCUSSION: Red blood cells were able to be efficiently recovered from clotted blood. Imaging studies did not show any evidence of red blood cells trapped within fibrin mesh.

Evaluation of Silicon Phthalocyanine 4 Photodynamic Therapy Against Human Cervical Cancer Cells In Vitro and in Mice.

BACKGROUND: Cervical cancer is the second most common cancer in women worldwide [1]. Photodynamic therapy has been used for cervical intraepithelial neoplasia with good responses, but few studies have used newer phototherapeutics. We evaluated the effectiveness of photodynamic therapy using Pc 4 in vitro and in vivo against human cervical cancer cells. METHODS: CaSki and ME-180 cancer cells were grown as monolayers and spheroids. Cell growth and cytotoxicity were measured using a methylthiazol tetrazolium assay. Pc 4 cellular uptake and intracellular distrubtion were determined. For in vitro Pc 4 photodynamic therapy cells were irradiated at 667nm at a fluence of 2.5 J/cm2 at 48 h. SCID mice were implanted with CaSki and ME-180 cells both subcutaneously and intracervically. Forty-eight h after Pc 4 photodynamic therapy was administered at 75 and 150 J/cm2. RESULTS: The IC50s for Pc 4 and Pc 4 photodynamic therapy for CaSki and ME-180 cells as monolayers were, 7.6µM and 0.016µM and >10µM and 0.026µM; as spheroids, IC50s of Pc 4 photodynamic therapy were, 0.26µM and 0.01µM. Pc 4 was taken up within cells and widely distributed in tumors and tissues. Intracervical photodynamic therapy resulted in tumor death, however mice died due to gastrointestinal toxicity. Photodynamic therapy resulted in subcutaneous tumor death and growth delay. CONCLUSIONS: Pc 4 photodynamic therapy caused death within cervical cancer cells and xenografts, supporting development of Pc 4 photodynamic therapy for treatment of cervical cancer. Support: P30-CA47904, CTSI BaCCoR Pilot Program.