In Vitro Enzymatic Degradation of Tissue Grafts and Collagen Biomaterials by Matrix Metalloproteinases: Improving the Collagenase Assay

Abstract

Matrix metalloproteinase-1 and -8 are active during the wound healing and remodelling processes, degrading native extracellular matrix and implantable devices. However, traditional in vitro assays utilize primarily matrix metalloproteinase-1 to mimic the in vivo degradation microenvironment. Herein, we assessed the influence of various concentrations of matrix metalloproteinase-1 and 8 (50, 100, and 200 U/mL) as a function of pH (5.5 and 7.4) and time (3, 6, 9, 12, and 24 h) on the degradation profile of three tissue grafts (chemically cross-linked Permacol, nonchemically crosslinked Permacol and nonchemically cross-linked Strattice) and a collagen biomaterial (nonchemically cross-linked collagen sponge). Chemically cross-linked and nonchemically cross-linked Permacol samples exhibited the highest resistance to enzymatic degradation, while nonchemically cross-linked collagen sponges exhibited the least resistance to enzymatic degradation. Qualitative and quantitative degradation analysis of all samples revealed a similar degradation profile over time, independently of the matrix metalloproteinase used and its respective concentration and pH. These data indicate that matrix metalloproteinase-1 and matrix metalloproteinase-8 exhibit similar degradation profile in vitro, suggesting that matrix metalloproteinase-8 should be used for collagenase assay.

Generation, Endothelialization, and Microsurgical Suture Anastomosis of Strong 1-mm-Diameter Collagen Tubes

“…Nevertheless, it is difficult to extrapolate from collagenase digestion rates to in vivo stability; we note that other studies of crosslinked collagenbased scaffolds have obtained similar mass loss rates. 27 Future studies with long-term in vivo implantation will be needed to better assess the stability of our crosslinked tubes.…”

The Collagen Suprafamily: From Biosynthesis to Advanced Biomaterial Development

“…In vitro enzymatic degradation of collagen‐based devices by matrix metalloproteinases, usually MMP‐1, allows investigation of the stability of the devices . However, MMP‐1 preferentially cleaves collagen type III, as opposed to MMP‐8, which is the predominant collagenase present in normal wound healing and degrades collagen type I more efficiently than MMP‐1 . MMP‐1, 2, 8, 13, and 14 are capable of hydrolyzing collagen types I, II, and III, while MMP‐3 and 9 are unable to degrade tropocollagen …”

Biomanufacturing Seamless Tubular and Hollow Collagen Scaffolds with Unique Design Features and Biomechanical Properties

“…In contrast to the hydrolytic degradation of PGA/PLGA, collagen mostly degrades in vivo via MMPs‐mediated enzymatic mechanism . We anticipate that collagen scaffolds will actively participate in dynamic interactions between cells and the regenerated tissues; therefore the fate of collagen scaffolds will depend on cross‐linking as well as tissue‐specific mechanical environment that are known to impact MMP‐mediated collagen degradation and remodeling in vivo . Thus, given their strength and compliance values ( Table 1 ), it is possible to develop tissue target‐specific collagen‐based scaffolds, which may demonstrate superior applicability as a scaffold material compared to previously investigated materials.…”