Small intestinal submucosa: a substrate for in vitro cell growth

Abstract

The extracellular matrix (ECM) of the small intestinal submucosa (SIS) was harvested by removing the superficial layers of the mucosa and the external muscular layers. The remaining 80 microns thick sheet was disinfected and sterilized by methods which removed all cellular components. The SIS-ECM, retaining its native 3-dimensional microarchitecture and composition, was evaluated for its ability to support in vitro cell growth. Six separate cell types were seeded either alone or in coculture with other cells upon this matrix, grown in selected media, a examined daily for time periods ranging from 48 h to 2 weeks. The six cell types tested were NIH Swiss mouse 3T3 fibroblast, NIH 3T3/j2 fibroblasts, primary human fibroblasts, primary human keratinocytes, human microvascular endothelial cells (HMECs), and an established rat osteosarcoma (ROS) cell line. All cell types showed the ability to attach a proliferate. All fibroblast cell line and the keratinocytes proliferated and/or migrated into the 3-dimensional scaffold of the SIS matrix. The ROS cells and the HMECs were confined in their growth pattern to the surface of the matrix. Coculturing of NIH 3T3/J2 fibroblasts and primary human keratinocytes resulted in a distinctive spatial orientation of the two cell types. The fibroblast populated the mid-substance of the 3-dimensional matrix and the keratinocytes formed an epidermal structure with rete ridge-like formation and stratification when the composite was lifted to an air liquid interface in culture. In summary, SIS provides a substratum with a 3-dimensional scaffold that allows for cell migration and spatial organization. The substratum is suitable for in vitro studies of the interaction between epithelial or mesenchymal cells and a naturally occurring extracellular matrix.

ECM-based triple layered scaffolds for vascular tissue engineering

“…Recently, layered vascular grafts have been prepared using decellularized small intestine submucosa (SIS) in combination with arterial elastin and cellularized collagen gels (4,36). A wide variety of constructs with a predefined layering and whose size can be tailored on the dimension of blood vessels to be replaced was achieved using this sandwich technique.…”

Nanostructured materials for applications in drug delivery and tissue engineering

“…Natural materials include collagen [131], silk protein [132], Matrigel [133], small intestinal submucosa (SIS) [134], agarose [135], alginate [136] and chitosan [137]. Although these materials show promise in tissue repair, critical issues regarding biocompatibility, mechanical properties and degradation cannot be neglected.…”

Use of a bioscaffold to improve healing of a patellar tendon defect after graft harvest for ACL reconstruction: A study in rabbits

“…Our findings suggest that the luminal side of SIS indeed acted as a barrier to limit cellular infiltration, while the abluminal side of SIS helped cellular proliferation and matrix production. 25,26 The lack of adhesion between the neo-PT and fat pad is of clinical relevance, as the maintenance of patellofemoral kinematics and knee mobilization may be achieved after BPTB graft harvest. Furthermore, the maintenance of relative motion between the PT and fat pad could provide the needed mechanical stimuli for PT healing while concomitantly reducing the deterioration of the remaining medial and lateral portions of the PT.…”

“…24 Further, SIS allows cellular infiltration only through its abluminal side, and thus, can prevent adhesion formation on its luminal side. 25,26 When a single layer of SIS was applied to a gap injury of the rabbit medial collateral ligament (MCL), the healing ligament had better collagen fiber alignment with larger collagen fibril diameters. 22,23 Also, the mechanical properties of the healing MCL were significantly improved with a 33% increase in tangent modulus and a 50% increase in tensile strength.…”

“…Using the right knee of the New Zealand White rabbit as a model, one layer of SIS was placed between the PT defect and the fat pad, and a second layer was placed over the anterior surface of the defect. As SIS has been shown to contain growth factors which can cause chemotaxis and to possess a preferential alignment of collagen fibers which may provide contact guidance to healing tissues, [24][25][26][27] the experiment was designed to test the hypothesis that SIS can cause increased cell proliferation and matrix production as well as improved matrix organization and biomechanical properties of the neo-PT. Secondly, based on the previous literature showing the ability of SIS to prevent cellular infiltration on its luminal side, 25,26 the SIS can serve as a natural barrier in order to limit cellular and vascular infiltration from the fat pad, thus limiting adhesion formation.…”