Extracellular Matrix Digestion Products for Tissue Engineering

Facing the medical needs regardless tissue engineering, we´re trying to develop an injectable hydrogel from digested ECMs. The digestion protocol was modified from [1] using trypsin, pepsin and collagenase. Glycosaminoglycan (GAGs) and peptides quantification were performed after continuous and stepwise digestion, showing that regardless the substrate and enzyme, the stepwise process yields a higher amount of GAGs and peptides, so a less degraded product. ECMs were also analyzed by proteomics, as in [2], to identify its composition. For tendon ECM: cadherin EGF LAG seven-pass G-type receptor 1, leucine rich-repeat containing 9, dynein axonemal heavy-chain and collagen alpha-1 (IV); pericardium ECM: folliculin interactin protein 2, TGF-beta receptor type-1, fibrillin-1, periostin, collagen alpha-1 (XII) chain, zinc-finger C2H2-type, histone H4, prolargin, collagen alpha-2 (1) chain, collagen type VI alpha-1 chain, lumican, annexin A5, anion exchange protein, olfactomedin like, collagen type VI alpha-3 chain, collagen type VI alpha-2 chain and protein kinase C-delta type; bone ECM: complement C3, collagen alpha-2(1) chain; ring finger protein 187 and ciliary rootlet coiled-coil. After proteomic, we verified that pericardium ECM has a highly diverse pool of proteins, differently from the other ECM types. Therefore, it is expected that after in vitro assays with stem cells, samples treated with pericardium ECM digested in stages might show better results for tissue regeneration.