EVALUATION OF DURABILITY IN BOVINE PERICARDIUM HEART VALVES SUBJECT TO CHEMICAL PROCESS DESCELULARIZAO

Angela M. Peruzzo, University Technological Federal do Paran, Curitiba, Francisco D. A. Costa, Irmandade da Santa Casa de Misericrdia de Curitiba, Carlos A. Laurindo, Joo G. Roderjan, Eduardo D. Vieira, Ricardo D. Torres, Paulo Soares, Pontifcia Universidade Catlica do Paran, Curitiba, Joo A. P. Setti, University Technological Federal do Paran, Curitiba. e-mail: angelaperuzzo@yahoo.com.br

INTRODUCTION
The heart valves can be classified into two categories according to the source: mechanics, manufactured with synthetic and biological material. The biological can be (heterologous): giver of different species. (E.g. bovine and porcine valves). Counterparts: donor of the same species. (Allografts or homografts can also prove useful) [MATHEUS, 2004]. Biological valves have a hydrodynamic performance better than the mechanical valves and also do not require continuous use of anticoagulant. However, they still need to improve the performance of its durability, this different chemical treatments as well as several tests have already been widely studied for a continuous improvement in valves that use biological membranes [COSTA, 1996]. There are now many ongoing research on the performance of bovine pericardium decellularized valves compared to others who also use the bovine pericardium with other types of chemical treatments. The decellularization of arrays are techniques to avoid the failure of the immune response induced valve allowing, in theory at least, improve the late survival of implanted valves [LOPES, 2009]. In order to have reliable results it is necessary to consider various factors that may interfere with the test as the operator training, the reliability of the equipment used for the execution of the tests and criteria for sampling which offer security in the final result, among others [PINTO, 2000]. The test will check if the decellularized pericardium gets minimal results stipulated in standard for the manufacture of heart valves, once your immune behavior tends to delay your implant after via useful. [LOPES, 2009].

MATERIALS AND METHODS

The method used for the fabrication of bovine pericardium heart valves of the control group and the test group was the same, changing only the chemical steps in the process to obtain the comparative results of different treatments. Preparation of the samples. Control group: the bovine pericardium control group underwent conventional treatment with glutaraldehyde [SUBIA, 1967] and used glutamic acid [FERREIRA, 2005] to decrease on calcification. Group test: To test the Group bovine pericardium was used the chemical treatment following the decellularization Protocol with valves, sodium dodecil sulfate (SDS), followed by fixing and sterilization with glutaraldehyde. In both groups the valves were stored in isotonic saline buffered and preserved with methyl paraben and propyl paraben. These chemical treatments aim to provide an adequate physical Constitution at bovine pericardium membrane, reduce its antigenicity for despicable levels, decreased calcification, as well as to ensure the final product sterility. Durability test. The test allows to estimate the lifetime of prostheses when placed in vivo. The equipment used was Vivitro Systems, model HCS 4991. The equipment can be seen in figures 1 and 2. Control group: Premium dentures Were used six 25 mm. The operating rate was 2000 cycles and closing pressure average was 100 5 mmHg. Group test: Tested on the same equipment, using three valves decellularized with 23 mm with operating frequency of 2000 cycles per minute and the closing pressure average 100 5 mmHg. Table 1. Results of durability tests Valves tested 01 control valve 05 valve controls 03 decellularized valves Durability 550 million cycles 800 million cycles 447 million cycles

Fig. 1. fatigue Simulator front view

Fig. 2. fatigue Simulator top view

RESULTS AND DISCUSSION

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In preliminary trials where they were tested six 25 mm Premium dentures, all exceeded the value of 200 million cycles accumulated, minimum value laid down in the standard ABNT ISO ISO 5840-5850-1999, and 2005. Being that the first failure occurred in a prosthesis with 550 million cycles. The other continued the test up to accumulate prostheses 800 million cycles, without any sign of failure as shown in dissertation performed by Laurindo, C.A., 2008. In the test group, where they were tested on the same equipment, three valves decellularized with 23 mm, was reached 447 million cycles and showed no sign of failure. The in vitro assays are performed in extreme conditions, and can estimate the useful life of the valve in vivo. Being that in vivo failure not valve gives only by physical factors, but also by biological factors inherent in each patient. Results of the tests are transcribed in table 1, above. Before arriving at a definitive conclusion and say that the results of obtained are within or not of the parameters required by the standardization is important to also assess some metrological aspects involved in determining a minimum measure or value obtained in an essay. At work was analyzed the pressure sensor error which presented a 0.83 calibration error and an expanded uncertainty of 2.24, indicated in the calibration report, which did not interfere in the final outcome of the evaluation, because in standard asks if a minimum pressure difference across the aortic valves closed 90 mmHg with a measurement accuracy of at least 5 mmHg. The same is true with the assessment of the relative standard uncertainty of the module indicated by R (T) = 2.56 mmHg/mV/v. the indicated module on standard uncertainty means indicated in the manual of the equipment [Albertazzi, 2008]. Then the value obtained from 447 million cycles for the test carried out tends to be considered suitable for decellularized bovine pericardium heart valves.

CONCLUSION
With the results obtained for the value of 447 million cycles in vivo was estimated a value of approximately 11 years. According to studies carried out by Costa, also concluded that the durability of decellularized reached similar results the valves made bovine pericardium with decellularized not. [COSTA, 2005]. The test is only one of numerous factors to be evaluated so that the process of decellularization in pericardium cattle can be considered suitable for the manufacture of heart valves. Getting here a suggestion for new tests that can lead to a definitive conclusion to the use or non-use of the technique used.
Referncias: A National Standars Institute, Inc. Association for the Advancement of Medical Instrumentation, International Standardization Organization 5840 (2005), Cardiovascular
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