Effects of sterilization on an extracellular matrix scaffold: Part I. Composition and matrix architecture

Abstract

The impact of peracetic acid (PAA), lyophilization, and ethylene oxide (EO) sterilization on the composition and three dimensional matrix structure of small intestinal submucosa (SIS), a biologic scaffold used to stimulate the repair of damaged tissues and organs, was examined. Fibronectin and glycosaminoglycans are retained in SIS following oxidation by peracetic acid and alkylation using ethylene oxide gas. Significant amounts of FGF-2 are also retained, but VEGF is susceptible to the effects of PAA and is dramatically reduced following processing. Further, matrix oxidation, lyophilization, and sterilization with EO can be performed without irreversibly collapsing the three dimensional structure of the native SIS. These structural features and growth promoting extracellular matrix constituents are likely to be important variables underlying cellular attachment, infiltration and eventual incorporation of SIS into healing host tissues.

Development and Characterization of Acellular Extracellular Matrix Scaffolds from Porcine Menisci for Use in Cartilage Tissue Engineering

“…Our studies also found that the collagen content was half of that of fresh menisci when treated with acetic acid for 2 h. These results indicate that acetic acid is not an ideal decellularization reagent due to its negative influence on collagen. In addition, peracetic acid is a common decellularization agent for porcine SIS and layers of the urinary bladder, 52,53 and it was reported to be highly efficient at removing residual nucleic acids from thin tissues with minimal effects on the ECM composition at concentrations of 0.10-0.15%. 30 However, here 0.15% peracetic had a limited decellularization effect on menisci in the first 2 h; furthermore, 15% peracetic acid treatment for 10 h decreased the DNA content to 4.68% -0.01%; while the amounts of collagen and GAG were respectively reduced to 44.98% -3.00% and 19.04% -4.01% compared to fresh menisci.…”

Tracheal Tissue Engineering

“…Through DSC analysis it also became clear that PAA treatment has a much more marked effect than originally anticipated. This contradicts existing literature [185,186,187] and is yet another testament to the importance of increasingly sophisticated analysis techniques, especially those capable of bulk tissue analysis. Through quantification of total enthalpy, a measure of overall thermal stability and higher order structure, we were able to determine that PAA significantly reduces thermal stability compared to all non-PAA groups.…”

“…Previous work has shown that PAA effectively removes cells and remnant DNA while having a minimal effect on ECM composition and structure [185,186,187]. However, given the broad applicability and success in the decellularization of membranes and thin planar tissues, there is surprisingly little quantitative data on PAA's benefit when compared to detergent based decellularization in larger, denser tissues.…”

“…We also tested per-acetic acid (PAA), since previous work had shown that PAA effectively removes cells and remnant DNA while having a minimal effect on ECM composition and structure [185,186,187].…”

Tissue engineering with gellan gum

“…Various techniques are in use for this purpose including autoclaving [34][35][36] , ethylene oxide treatment 37 , microfiltration 38 , and antibiotic supplements 39 . Although no studies directly compare these methods with respect to their impacts on GG, equivalent investigations using other polysaccharides [40][41][42][43] and ECM materials 40,44,45 provide a good indication of the likely effects on GG. In general, these studies report that common sterilisation procedures are generally effective at removing or deactivating contaminants without loss of the biological activity.…”