Proteomic Analysis To Uncover Variability And Conserved Proteins In The Decellularization Process Of Porcine And Ovine Semilunar Valves
Renata C. C. Freitas, PhD1, Cassandra L. Clift, Ph.D.2, Kathryn G. Henshaw, B.S.1, Angelina J. Joyce, B.A.1, Abhijeet Sonawane, Ph.D.2, Sasha A. Singh, Ph.D.2, Elena Aikawa, M.D., Ph.D.2, John E. Mayer Jr, M.D.1.
1Boston Children's Hospital, Boston, MA, USA, 2Brigham and Women's Hospital, Boston, MA, USA.
Objective(s): The extracellular matrix (ECM) protein composition of heart valves is thought to be conserved across various mammalian species, with immunogenicity caused by cellular components. Proteomic analysis of cardiac valve tissue offers insights into the clinical limitations of heterograft valve devices and may provide guidance for tissue engineering approaches to congenital heart disease. This study assessed proteomic variations between native and decellularized porcine and ovine leaflets.
Methods: Pulmonary valve leaflets were excised from porcine (N=4) and ovine (N=3) hearts. Leaflets were decellularized using NaOH, ionic detergents and nucleases, then analyzed histologically to assess decellularization. Proteomic analysis was conducted using liquid chromatography mass spectrometry.
Results: Proteomic analysis of native porcine and ovine samples revealed significant interspecies variability in individual protein abundances (p<0.05). Unique proteins were identified in each species (392 in porcine/541 in ovine) from a total of >3,000 proteins. Decellularization reduced the total number of proteins compared to native leaflets and enriched ECM and matricellular proteins. Despite decellularization, significant differences persisted between ovine and porcine tissues, indicating discrete species-specific matrix components remain. Following xenograft transplant, species-specific ECM-targeted proteomics analysis can distinguish between native ECM proteins and those deposited by seeded cells during tissue-engineered heart valve implantation. Conclusions: Porcine and ovine leaflets had significant interspecies proteomic variations, which were reduced, but not eliminated, by decellularization. Decellularization preserved some ECM components but the effects on immunogenicity require further study. These interspecies variations have important potential implications for both heterograft and tissue engineered valve materials.
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