Injectable small intestinal submucosa is co-cultured with adipose-derived mesenchymal stem cells in vitro

doi:10.3969/j.issn.2095-4344.2016.38.016

Abstract

Abstract:

BACKGROUND: The decellularized porcine small intestinal submucosa is a kind of bioactive extracellular matrix, which is mainly composed of collagen, glycoprotein, proteoglycan and rich in collagen, glycosaminoglycan and various growth factors, and these components play an important role in promoting the differentiation and proliferation of tissue cells.
OBJECTIVE: To prepare the injectable small intestinal submucosa and to investigate its co-culture with rat adipose-derived mesenchymal stem cells in vitro.
METHODS: The injectable small intestinal submucosa and rat adipose-derived stem cells were prepared. Cell counting kit-8 test for cell proliferation: Passage 3 adipose-derived stem cells were seeded onto the injectable small intestinal submucosa (experimental group) and cells cultured under normal condition as control group. The cell proliferation was observed at 1, 3, 5 and 7 days of incubation. Live/dead staining test for the survival of cells: Passage 3 adipose-derived stem cells were respectively cultured in the injectable small intestinal submucosa extracts (experimental group) and complete culture medium (control group). Cell survival was determined at 1, 3, 5 and 7 days of culture.
RESULTS AND CONCLUSION: Scanning electron microscope oval and strip adipose- derived stem cells adhered onto the material. The absorbance values in the experimental group were higher than those in the control group at 1 and 5 days of incubation (P < 0.05). Cell survival: The number of cells appeared to be in a rising trend with time in both two groups; after 1-day co-culture, all cells in the two groups survived. Then dead cells appeared in both two groups, showing no significant difference. These results show that the injectable small intestinal submucosa exhibits a good cytocompatibility.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

Key words: Extracellular Matrix, Stem Cells, Tissue Engineering

CLC Number:

R318

Guo Xing, Zhou Hong, Li Dan, Gao Xiao-chun, Dai Lei, Huang Hai-jun, Tan Mei-yun.

Injectable small intestinal submucosa is co-cultured with adipose-derived mesenchymal stem cells in vitro

[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(38): 5730-5736.

Figures/Tables 6

References

[1]Wilson A,Butler PE,Seifalian AM.Adipose-derived stem cells for clinical applications: A review.Cell Prolif. 2011; 44(1):86-98.
[2]Hong SJ,Jia SX,Xie P,et al.Topically delivered adipose derived stem cells show an activated-fibroblast phenotype and enhance granulation tissue formation in skin wounds. PloS One.2013;8(1):e55640.
[3]Dong Y,Hassan WU,Kennedy R,et al.Performance of an in situ formed bioactive hydrogel dressing from a peg-based hyperbranched multifunctional copolymer.Acta Biomaterialia. 2014;10(5):2076-2085.
[4]Gugerell A,Neumann A,Kober J,et al.Adipose-derived stem cells cultivated on electrospun l-lactide/glycolide copolymer fleece and gelatin hydrogels under flow conditions-aiming physiological reality in hypodermis tissue engineering. Burns.2015;41(1):163-171.
[5]Haubner F,Gassner HG.Potential of adipose-derived stem cells concerning the treatment of wound healing complications after radiotherapy. HNO. 2015;63(2): 111-117.
[6]Ozpur MA,Guneren E,Canter HI,et al.Generation of skin tissue using adipose tissue-derived stem cells. Plast Reconstr Surg.2016;137(1):134-143.
[7]Shalaby SM,Sabbah NA,Saber T,et al.Adipose-derived mesenchymal stem cells modulate the immune response in chronic experimental autoimmune encephalomyelitis model.IUBMB Life.2016;68(2): 106-115.
[8]Zuk PA,Zhu M,Mizuno H,et al.Multilineage cells from human adipose tissue: Implications for cell-based therapies.Tissue Eng.2001;7(2):211-228.
[9]Park BS,Jang KA,Sung JH,et al.Adipose-derived stem cells and their secretory factors as a promising therapy for skin aging.Dermatol Surg. 2008;34(10):1323-1326.
[10]Taha MF,Hedayati V.Isolation, identification and multipotential differentiation of mouse adipose tissue-derived stem cells.Tissue Cell. 2010;42(4): 211-216.
[11]Gimble JM,Katz AJ,Bunnell BA.Adipose-derived stem cells for regenerative medicine.Circ Res.2007;100(9): 1249-1260.
[12]Yamane S,Iwasaki N,Majima T,et al.Feasibility of chitosan-based hyaluronic acid hybrid biomaterial for a novel scaffold in cartilage tissue engineering. Biomaterials. 2005;26(6):611-619.
[13]Lutolf MP,Hubbell JA.Synthetic biomaterials as instructive extracellular microenvironments for morphogenesis in tissue engineering.Nat Biotechnol. 2005;23(1):47-55.
[14]Shi L,Ronfard V.Biochemical and biomechanical characterization of porcine small intestinal submucosa (sis): A mini review.Int J Burns Trauma.2013;3(4): 173-179.
[15]Davies OG,Cooper PR,Shelton RM,et al.Isolation of adipose and bone marrow mesenchymal stem cells using cd29 and cd90 modifies their capacity for osteogenic and adipogenic differentiation.J Tissue Eng.2015;6:2041731415592356.
[16]Nie C,Yang D,Xu J,et al.Locally administered adipose-derived stem cells accelerate wound healing through differentiation and vasculogenesis.Cell Transplant.2011;20(2):205-216.
[17]Kang KN,Kim da Y,Yoon SM,et al.In vivo release of bovine serum albumin from an injectable small intestinal submucosa gel.Int J Pharm.2011;420(2): 266-273.
[18]Kim K,Kim MS.An injectable hydrogel derived from small intestine submucosa as a stem cell carrier.J Biomed Mater Res B Appl Biomater.2015.doi: 10.1002/jbm.b.33504. [Epub ahead of print]
[19]Guo X,Xia B,Lu XB,et al.Grafting of mesenchymal stem cell-seeded small intestinal submucosa to repair the deep partial-thickness burns.Connect Tissue Res. 2016:1-10.
[20]Cowles EA,Brailey LL,Gronowicz GA. Integrin-mediated signaling regulates ap-1 transcription factors and proliferation in osteoblasts. J Biomed Mater Res.2000;52(4):725-737.
[21]Andree B,Bar A,Haverich A,et al.Small intestinal submucosa segments as matrix for tissue engineering: Review.Tissue Eng Part B Rev.2013;19(4):279-291.
[22]杨浩,吴迪,李世和,等.猪小肠黏膜下层基质支架材料复合脂肪基质干细胞的生物相容性[J].中国组织工程研究与临床康复,2010,14(3):415-418.
[23]房艳,倪伟民,单伟,等.海绵状的小肠粘膜下层促进成骨样细胞增殖分化[J].中国生物工程杂志,2013,33(6):18-23.
[24]田伟,王效杰.猪小肠黏膜下层海绵的三维重建、表面修饰和细胞黏附性[J].中国老年学杂志, 2014,34(14):3949- 3951.
[25]Choi JW,Park JK,Chang JW,et al.Small intestine submucosa and mesenchymal stem cells composite gel for scarless vocal fold regeneration.Biomaterials. 2014;35(18):4911-4918.
[26]Zonari A,Martins TM,Paula AC,et al. Polyhydroxybutyrate-co-hydroxyvalerate structures loaded with adipose stem cells promote skin healing with reduced scarring.Acta Biomaterialia. 2015;17: 170-181.
[27]Zeng Y,Zhu L,Han Q,et al.Preformed gelatin microcryogels as injectable cell carriers for enhanced skin wound healing.Acta Biomaterialia. 2015;25: 291-303.
[28]Kono H,Teshirogi T.Cyclodextrin-grafted chitosan hydrogels for controlled drug delivery.Int J Biol Macromol. 2015;72:299-308.
[29]Kwon JS,Yoon SM,Shim SW,et al.Injectable extracellular matrix hydrogel developed using porcine articular cartilage.Int J Pharm.2013;454(1):183-191.
[30]Naderi-Meshkin H,Andreas K,Matin MM,et al. Chitosan-based injectable hydrogel as a promising in situ forming scaffold for cartilage tissue engineering. Cell Biol Int.2014;38(1):72-84.