Tubular scaffolds of gelatin and poly(ε-caprolactone)-block-poly(γ-glutamic acid) blending hydrogel for the proliferation of the primary intestinal smooth muscle cells of rats

Biomed Mater. 2013 Dec;8(6):065002. doi: 10.1088/1748-6041/8/6/065002. Epub 2013 Nov 14.

Abstract

The proper regeneration of intestinal muscle for functional peristalsis is the most challenging aspect of current small intestine tissue engineering. This study aimed to fabricate a hydrogel scaffold for the proliferation of intestinal smooth muscle cells (ISMCs). Tubular porous scaffolds of 10-20 wt% gelatin and 0.05-0.1 wt% poly(ε-caprolactone)-block-poly(γ-glutamic acid) blending hydrogel were cross-linked by carbodiimide and succinimide in an annular space of a glass mold. The scaffolds with higher gelatin contents degraded slower in the phosphate buffer solution. In rheological measurements, the hydrated scaffolds were elastic (all tangent delta <0.45); they responded differentially to frequency, indicating a complete viscoelastic property that is beneficial for soft tissue regeneration. Isolated rat ISMCs, with the characteristic biomarkers α-SMA, calponin and myh11, were loaded into the scaffolds by using either static or centrifugal methods. The average cell density inside the scaffolds increased in a time-dependent manner in most scaffolds of both seeding groups, although at early time points (seven days) the centrifugal seeding method trapped cells more efficiently and yielded a higher cell density than the static seeding method. The static seeding method increased the cell density from 7.5-fold to 16.3-fold after 28 days, whereas the centrifugal procedure produced a maximum increase of only 2.4-fold in the same period. In vitro degradation data showed that 50-80% of the scaffold was degraded by the 14th day. However, the self-secreted extracellular matrix maintained the integrity of the scaffolds for cell proliferation and spreading for up to 28 days. Confocal microscopic images revealed cell-cell contacts with the formation of a 3D network, demonstrating that the fabricated scaffolds were highly biocompatible. Therefore, these polymeric biomaterials hold great promise for in vivo applications of intestinal tissue engineering.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / metabolism
  • Animals
  • Biocompatible Materials / chemistry*
  • Calcium-Binding Proteins / metabolism
  • Calponins
  • Cell Count
  • Cell Proliferation
  • Elasticity
  • Gelatin
  • Humans
  • Hydrogels
  • Intestine, Small / cytology
  • Intestine, Small / physiology
  • Materials Testing
  • Microfilament Proteins / metabolism
  • Microscopy, Electron, Scanning
  • Models, Animal
  • Myocytes, Smooth Muscle / cytology*
  • Myocytes, Smooth Muscle / physiology
  • Myosin Heavy Chains / metabolism
  • Polyesters / chemistry
  • Polyglutamic Acid / analogs & derivatives
  • Polyglutamic Acid / chemistry
  • Rats
  • Regeneration / physiology
  • Short Bowel Syndrome / physiopathology
  • Short Bowel Syndrome / therapy
  • Tissue Engineering / methods*
  • Tissue Scaffolds / chemistry*
  • Viscosity

Substances

  • Actins
  • Biocompatible Materials
  • Calcium-Binding Proteins
  • Hydrogels
  • Microfilament Proteins
  • Polyesters
  • poly(gamma-glutamic acid)
  • smooth muscle actin, rat
  • polycaprolactone
  • Polyglutamic Acid
  • Gelatin
  • Myosin Heavy Chains