Bioengineering functional smooth muscle with spontaneous rhythmic contraction in vitro

Abstract

Oriented smooth muscle layers in the intestine contract rhythmically due to the action of interstitial cells of Cajal (ICC) that serve as pacemakers of the intestine. Disruption of ICC networks has been reported in various intestinal motility disorders, which limit the quality and expectancy of life. A significant challenge in intestinal smooth muscle engineering is the rapid loss of function in cultured ICC and smooth muscle cells (SMC). Here we demonstrate a novel approach to maintain the function of both ICC and SMC in vitro. Primary intestinal SMC mixtures cultured on feeder cells seeded electrospun poly(3-caprolactone) scaffolds exhibited rhythmic contractions with directionality for over 10 weeks in vitro. The simplicity of this system should allow for wide usage in research on intestinal motility disorders and tissue engineering, and may prove to be a versatile platform for generating other types of functional SMC in vitro.

Understanding the Biology of Human Interstitial Cells of Cajal in Gastrointestinal Motility

“…When primary cultured mouse ‘intestinal muscularis complexes’ (containing Kit + cells) were combined with electrospun poly-caprolactone scaffold sheets in vitro, spontaneous contractions were observed [ 65 ]. Similar findings occurred when electrospun poly-caprolactone scaffold sheets were seeded with STO feeder cells and primary MACS-purified mouse Kit + ICC [ 66 ], thus demonstrating attachment and alignment onto the scaffold sheets in vitro. In addition, when a primary mouse intestinal smooth muscle cell mixture was cultured on STO-seeded electrospun poly-caprolactone scaffold sheets, rhythmic contractions developed, and Kit + Ano1 + cells were observed after 10 weeks.…”

“…This included spontaneous contractions and the presence of gut cell types including Kit + cells. To further maintain the functionality of ICC in vitro, STO (SIM, Sandos Inbred Mouse) embryonic fibroblast feeder cells have been co-cultured with enriched murine Kit + ICC (purified from primary mouse intestinal smooth muscle cell mixture through the use of magnetic-activated cell sorting and an anti-Kit antibody) [ 66 ]. Intriguingly, this model maintained contractility and contained Kit + Ano1 + cells for up to 14 days.…”

Biomaterial-Based Approaches for Regeneration of Periodontal Ligament and Cementum Using 3D Platforms

“…In addition to the soft lithographic strategy, the electrospinning fabrication technique has been widely utilized to manufacture random or highly aligned nano-/micro-fibrous membranes in dental tissue engineering applications such as periodontal regeneration [50,51,52] or pulp regeneration [53,54,55]. In particular, the electrospun 2D membranes with unidirectional alignments or controllable-aligned patterns of nano-fibers were investigated for specific cell orientations and migrations to regulate cellular responses to designed microenvironments [56,57]. However, there is the limitation to create 3D directional internal architectures to regulate cell or tissue orientations with structural similarities to PDLs because periodontal connective tissues have the spatially perpendicular or oblique angulations to tooth-root surfaces as described above [51,58].…”

Development of a conductive biocomposite combining graphene and amniotic membrane for replacement of the neuronal network of tissue-engineered urinary bladder

“…There is a shortage of literature demonstrating contraction of tissue-engineered grafts in vitro, including the lack of a reference protocol. Kobayashi et al determined that intestinal SMS seeded electrospun poly(3-caprolactone) scaffolds exhibited rhythmic contractions resulting in biomaterial contraction 42 . Krueger et al reported the ability of muscle cells, forming myotubes seeded on graphene foam to mediate consolidated micromovements 43 .…”