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

• Published in: Biomedical materials (Bristol) Vol. 8; no. 6; p. 065002
• Main Authors: Jwo, Shyh-Chuan, Chiu, Chu-Hua, Tang, Shye-Jye, Hsieh, Ming-Fa
• Format: Journal Article
• Language: English
• Published: England IOP Publishing 01.12.2013
• Subjects: Actins - metabolism; Animals; Biocompatible Materials - chemistry; Calcium-Binding Proteins - metabolism; Calponins; Cell Count; Cell Proliferation; Elasticity; Gelatin; Humans; Hydrogels; intestinal smooth muscle cells; 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; poly ( -caprolactone)-poly (γ-glutamic acid) block copolymer; Polyesters - chemistry; Polyglutamic Acid - analogs & derivatives; Polyglutamic Acid - chemistry; Rats; Regeneration - physiology; Short Bowel Syndrome - physiopathology; Short Bowel Syndrome - therapy; small intestine tissue engineering; Tissue Engineering - methods; Tissue Scaffolds - chemistry; Viscosity
• ISSN: 1748-6041; 1748-605X
• DOI: 10.1088/1748-6041/8/6/065002

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( -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.