Ultrastructural analysis and residual DNA evaluation of rabbit vein scaffold

Abstract

Purpose: To investigate the ultrastructural characteristics and analysis of residual DNA in scaffold models, produced with decellularized vena cava in an experimental model with rabbits. Methods: Three groups were created for ultrastructural and residual DNA analysis: group 1 -control, consisting of samples of vena cava in natura; group 2 -SD, consisting of vein fragments submitted to 2% sodium deoxycholate decellularization by shaking (160rpm -Shaker News Brunswick Scientific ® ) for 1 hour at controlled temperature shaker at 37°C; group 3 -SDS, consisting of vein fragments submitted to 1% sodium dodecyl sulfate decellularization under the same previous condition, for 2 hours. Results: The ultrastructural matrix of the blood vessel maintained its vintegrity after either decellularization models. The results of the two quantification methods demonstrated a significant decrease in the DNA content of the decellularized vena cava samples as compared to the control samples and, differed statistically from each other, p <0.05. Conclusion: The 2% DS protocol for vein decellularization, in this experimental model, was considered the best protocol because it presented less amount of residual DNA without causing substantial destruction of the extracellular matrix.

Urinary Bladder Patch Made with Decellularized Vein Scaffold Seeded with Adipose-Derived Mesenchymal Stem Cells: Model in Rabbits

“…The IVCs were fragmented into 1 cm length segments and processed with the standardized decellularization protocol using 1% sodium dodecyl sulfate (SDS, Sigma-Aldrich, Santo André, SP, Brazil) with a 2 h, 37 °C incubation on a shaker ( Supplemental File S1 ). The decellularization process aims to remove all cellular and nuclear material, including antigens, while preserving the structural and mechanical integrity and chemical properties of the extracellular matrix [ 12 , 13 ]. The decellularized fragments were maintained in a sterile solution containing antibiotic ciprofloxacin (Ciprofloxacin 10 mg/mL, Bayer, Paulina, SP, Brazil) and antifungal amphotericin B (Amphotericin B 20 mg/mL, Geolab, Anápolis, GO, Brazil) at 4 °C for 7 days.…”

“…Furthermore, the physicochemical signals remain intact for regulating cell adhesion, proliferation, and differentiation, thus, providing a superior substrate for MSC seeding [ 7 ]. The scaffold produced through venous decellularization has biomechanical properties sufficient to resist the stresses of the urinary system and offers an extracellular matrix favorable to the cell population [ 12 , 13 , 14 ]. Tissue engineering research for the production and replacement of urinary bladder tissue has grown in recent decades; however, the clinical applications have not been completely established [ 15 , 16 ].…”

Evaluation of Biointegration and Inflammatory Response to Blood Vessels Produced by Tissue Engineering—Experimental Model in Rabbits

“…A major challenge for TEBV to widely reach clinical practice is the production of a functional, resistant, and three-dimensional blood vessel [ 6 , 7 ]. For that, the basis of development requires that the scaffold (synthetic or biological) be sufficient to provide the necessary support for the implanted stem cells to differentiate, renew the extracellular matrix (ECM) and do not induce immunological reactions [ 8 , 9 , 10 ].…”