Effect of Heart Valve Decellularization on Xenograft Rejection.

OBJECTIVES: Endothelial cells harbor many antigenic determinants that may be targets for the immune system. The aim of this study was to determine the immunologic effects of decellularization, using 3 different methods, on xenograft rejection. MATERIALS AND METHODS: In a sterile plate containing phosphate-buffered saline, fresh sheep aortic heart valves were decellularized using 3 different enzymatic methods: with 900 µg/mL of collagenase at 40°C (method A), with 450 µg/mL of collagenase at 4°C (method B), and with 900 µg/mL of collagenase at 4°C (method C). Intact and decellularized valves were implanted subdermally into inbred male albino rabbits and extracted after 21 days (extra valve pieces were also extracted after 60 days, as control samples, for assessing chronic rejection). Valves were histologically analyzed for inflammatory cell infiltration. Subendothelial structure integrity was determined using surface electron microscope. RESULTS: No inflammatory cell infiltration was seen around the decellularized valve with method A, and no subendothelial structure change was observed by surface electron microscope. Infiltration of immune cells involved in rejection was not seen around valves decellularized with method B, although the subendothelial structure was relatively preserved and valve stiffness was increased. With method C, we observed a foreign body-type reaction around the intact valve and the decellularized valve. CONCLUSIONS: Method A is considered the optimal method of decellularization in our study, as this method significantly reduced the immune response to xenograft tissue, while maintaining subendothelial tissue.

Increased production of soluble vascular endothelial growth factors receptor-1 in CHO-cell line by using new combination of chitosan-protein lipid nanoparticles.

The soluble vascular endothelial growth factor receptor-1 (VEGFR1) or sFLT-1 has important role in antiangiogenesis. In this study, the increase expression and production of sFLT-1 fragment by newly designed ChPL-NPs nanoparticles (chitosan-protein lipid) using Chinese hamster ovary cell line (CHO) was evaluated. The assessment and purification of sFLT-1 were carried out by western blotting and fast protein liquid chromatography (FPLC). Thereafter, the angiostatic effect of gene transfer of sFLT-1 in Human umbilical vein endothelial cell line (HUVEC) was evaluated. Our results showed a significance rate of transfection with ChPL-NPs (80-85%) in comparison to standard lipofectamine(2000) (65-70%) (P < 0.05). The anti-angiogenic action of sFLT-1 was observed by in-vitro culture of recombinant protein (sFLT-1; 50 ng/ml) with HUVEC cell lines (5 × 10(6)). The ChPL-NPs nanoparticles can consider a potential carrier system for large scale production of sFLT-1, which ultimately may be use as therapeutic agent in targeting solid tumor tissues.

Histological assessment of pulp condition after apical vital root transection in one root of multirooted teeth in dogs: a preliminary study.

One of the most important aspects in surgery is the healing process after the periapical surgery. Past studies have shown occasional encounters with vital root resection and have noted varying degrees of pulpal response after root resection in periodontal disease. The purpose of this investigation was to observe the pulpal and periapical responses to intentional apical vital root transection in one root of multirooted teeth of German-Canadian dogs over a 6-month postoperative period. This is an experimental study performed on left maxillary and mandibular quadrants of four adult German-Canadian dogs after a 3- and 6-month period. Four teeth were assessed in each interval. One of the roots of multirooted teeth in the left quadrant of both maxillary and mandibular jaws was surgically transected. Tissue blocks were prepared by routine histological methods after 12 and 24 weeks after the surgery. The results showed a disruption of the normal pulpal architecture, with initial pulpal degeneration and subsequent early replacement by the periodontal ligament tissue after 24 weeks. Hypercementosis was seen around the apical portion of the root in all specimens. Pulpal regeneration was seen in the both upper and lower molars (p = 0.03). Resorption took place only in two specimens (p = 0.46). The inflammation in the 12th week was more than the 24th week. The pulp of multirooted teeth remains vital after transection of the apical part of the root in dogs. Longer follow-up periods are recommended because root canal therapy or extraction is indicated if resorption, necrosis, or ankylosis is seen.