Objective To investigate the biological and biomechanical characteristics of acellular porcine aortic valve with dye mediated photo oxidation so that a new and better bioprosthetic valve materials can be obtained. Methods Thirty porcine aortic valves were divided into three groups with random number table. Acellular valves (n=10) were stabilized by dye mediated photo oxidation in dye mediated photo oxidation group; acellular valves (n=10) were stabilized by glutaraldehyde in glutaraldehyde group; and acellular valves (n=10) were acellularized only in acellular valves group. Thickness, appearance, histology, water content, shrinkage temperature, breaking strength and soluble protein level of acellular porcine aortic in three groups were tested respectively. Results There were light blue, soft, flexible and unshrinking valves in dye mediated photo oxidation group. Compared to valves in glutaraldehyde group, valves in dye mediated photo oxidation group had lighter thickness（0.26±0.09mm vs. 0.38±0.08mm，Plt;0.05）, more water content（86.30%±4.03% vs. 71.10%±3.23%，Plt;0.05）, and lower shrinkage temperature （76.30±0.70℃ vs. 87.70±0.30℃，Plt;0.05）; while these indexes had no statistically significant differences compared to those in acellular valves group. At the same time, compared to valves in acellular valves group, valves in dye mediated photo oxidation group had more breaking strength（17.33±2.65 mPa vs. 9.11±0.95 mPa，Plt;0.05） and lower soluble protein level（0.039%±0.013% vs. 0.107%±0.024%，Plt;0.05）; while these indexes had no statistically significant differences compared to those in glutaraldehyde group. Conclusion Acellular porcine aortic valve stabilized by dye mediated photo oxidation has nice biological and biomechanical characteristics.
【Abstract】 Objective To investigate the feasibil ity of applying enzymatic method to prepare decellularizedporcine aorta and to evaluate its biomechanical properties, immunogenicity and cell compatibil ity. Methods 0.1% trypsin- 0.01% EDTA was appl ied to extract cells from porcine aorta under 37 continuously vibrating condition and its histology and microstructure were observed. The thickness, stress-strain curve, ultimate tension stress (UTS) and strain of failure (SOF) were compared before and after decellularization for 48, 96 and 120 hours under uniaxial tensile tests, respectively. The histological change was observed at 1, 3 and 6 weeks after the decellularized tissue was implanted subcutaneously in 3 dogs. According to the HE stains and a semi-quantitative Wakitani grading method, gross changes, category and amounts of infiltrated cells and neo-capillaries were compared between pre- and post-decellularization of porcine aortae. Endothel ial cells from canine external jugular vein were seeded onto the decellularized patches to observe the cell compatibil ity. Results Microscopy and electron microscopies examination identified that cell components was completely removed from the fresh porcine aorta and Masson’ strichrome showed that the structure of matrix (fibrins) was maintained intact at 96 hours using the decellularization method. There were no significant differences in the thickness, UTS and SOF between before and after decellularization (P gt; 0.05). However, The UTS values showed a decrease tendency and SOF showed an increase tendency. The stress-strain curve also verified a decrease tendency in mechanical intensity and an increase one in ductil ity after decellularization. After implanting the acellularized matrix subcutaneously in canine, moderately lymphocyte infiltration was seen at the 1st week and the infiltration was replaced by fibroblasts accompanied by neocapillary formation at the 6th week. A semi-quantity histological evaluation showed that there were differences in gross observation, category and the numbers of the infiltrated cells between decellularized and non-decellularized tissues（P lt; 0.05). A cell monolayer was identified by HE staining and scanning electron microscopywhen the endothel ial cells were seeded onto the inner luminal surface of the scaffold, al igned at the same direction on the whole. Conclusion The decellularized porcine aortic scaffold, prepared by trypsin-EDTA extraction under continuously vibrating condition, could meet the requirements of tissue-engineering graft in biomechanical properties, immunogenicity and cell compatibil ity.
In order to understand how the biomechanical properties of rabbit cornea change over time after corneal ablation, 21 healthy adult rabbits were used in this study, with the left eye as experimental side and the right eye as the control side. Firstly, a lamellar knife was used to remove a portion of the anterior corneal surface tissue (30%~50% of the original corneal thickness) from the left eye of each rabbit, as an animal model simulating corneal refractive surgery. Secondly, postoperative experimental rabbits were kept for one, three, or six months until being euthanized. Strip specimens were produced using their corneas in vitro to perform a uniaxial tensile test with an average loading-unloading rate of approximately 0.16 mm/s. Finally, the visco-hyperelastic material constitutive model was used to fit the data. The results showed that there was a significant difference in the viscoelastic parameters of the corneas between the experimental and the control eyes at the first and third postoperative months. There was a difference in tangential modulus between the experimental and the control eyes at strain levels of 0.02 and 0.05 at the third postoperative month. There was no significant difference in biomechanical parameters between the experimental and the control eyes at the sixth postoperative month. These results indicate that compared with the control eyes, the biomechanical properties of the experimental eyes vary over postoperative time. At the third postoperative month, the ratio of corneal tangential modulus between the experimental and the control eyes significantly increased, and then decreased. This work lays a preliminary foundation for understanding the biomechanical properties of the cornea after corneal refractive surgery based on rapid testing data obtained clinically.