PREPARATION AND BIOCOMPATIBILITY OF PORCINE SKELETAL MUSCLE ACELLULAR MATRIX FOR ADIPOSE TISSUE ENGINEERING_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

TIAN Chunxiang ^1,2 , FAN Xuejiao ^1,2 , CHEN Xiaohe ² , DENG Li ² , QIN Tingwu ² , LUO Jingcong ² , LI Xiuqun ² , LUuml ¹

1Department of Thyroid &;
Breast Surgery, 2State Key Laboratory of Biotherapy, Division of Stem Cell and Tissue Engineering, West China Hospital, Sichuan University, Chengdu Sichuan, 610041, P.R.China.;

Corresponding author：

Keywords：

Adi pose tissue engineering; Skeletal muscle; Acellular matrix; Human adi pose-derived stem cells; Scaffold material

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Objective Extracellular matrix is one of the focus researches of the adi pose tissue engineering. To investigate the appropriate method to prepare the porcine skeletal muscle acellular matrix and to evaluate the biocompatibility of the matrix. Methods The fresh skeletal muscle tissues were harvested from healthy adult porcine and were sl iced into
2-3 mm thick sheets, which were treated by hypotonic-detergent method to remove the cells from the tissue. The matrix was then examined by histology, immunohistochemistry, and scanning electron microscopy. The toxic effects of the matrix were tested by MTT. Human adi pose-derived stem cells (hADSCs) were isolated from adi pose tissue donated by patients with breast cancer, and identified by morphology, flow cytometry, and differentiation abil ity. Then, hADSCs of passage 3 were seeded into the skeletal muscle acellular matrix, and cultured in the medium. The cellular behavior was assessed by calcein-AM (CA) and propidium iodide (PI) staining at 1st, 3rd, 5th, and 7th days after culturing. Results Histology, immunohistochemistry, and scanning electron microscopy showed that the muscle fibers were removed completely with the basement membrane structure; a large number of collagenous matrix presented as regular network, porous-like structure. The cytotoxicity score of the matrix was grade 1, which meant that the matrix had good cytocompatibil ity. The CA and PI staining showed the seeded hADSCs had the potential of spread and prol iferation on the matrix. Conclusion Porcine skeletal muscle acellular matrix has good biocompatibility and a potential to be used as an ideal biomaterial scaffold for adi pose tissue engineering.

Citation： TIAN Chunxiang,FAN Xuejiao,CHEN Xiaohe,DENG Li,QIN Tingwu,LUO Jingcong,LI Xiuqun,LUuml,Qing. PREPARATION AND BIOCOMPATIBILITY OF PORCINE SKELETAL MUSCLE ACELLULAR MATRIX FOR ADIPOSE TISSUE ENGINEERING. Chinese Journal of Reparative and Reconstructive Surgery, 2012, 26(6): 749-754. doi: Copy

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1. Lutolf MP, Hubbell JA. Synthetic biomaterials as instructive extracellular microenvironments for morphogenesis in tissue engineering. Nat Biotechnol, 2005, 23(1): 47-55.
2. Gilbert TW, Sellaro TL, Badylak SF. Decellularization of tissues and organs. Biomaterials, 2006, 27(19): 3675-3683.
3. Borschel GH, Dennis RG, Kuzon WM Jr. Contractile skeletal muscle tissue-engineered on an acellular scaffold. Plast Reconstr Surg, 2004, 113(2): 595-602.
4. Valentin JE, Turner NJ, Gilbert TW, et al. Functional skeletal muscle formation with a biologic scaffold. Biomaterials, 2010, 31(29): 7475-7484.
5. Penolazzi L, Mazzitelli S, Vecchiatini R, et al. Human mesenchymal stem cells seeded on extracellular matrix scaffold: Viability and osteogenic potential. J Cell Physiol, 2012, 227(2): 857-866.
6. Zhu WD, Xu YM, Feng C, et al. Different bladder defects reconstructed with bladder acellular matrix grafts in a rabbit model. Urologe A, 2011, 50(11): 1420-1425.
7. Zhang X, Yang J, Li Y, et al. Functional neovascularization in tissue engineering with porcine acellular dermal matrix and human umbilical vein endothelial cells. Tissue Eng Part C Methods, 2011, 17(4): 423-433.
8. 张建, 吴英锋, 陈亮. 猪主动脉脱细胞基质的简化制备及生物学评价. 中国修复重建外科杂志, 2008, 22(3): 364-369.
9. Wainwright JM, Hashizume R, Fujimoto KL, et al. Right ventricular outflow tract repair with a cardiac biologic scaffold. Cells Tissues Organs, 2012, 195(1-2): 159-170.
10. Soto-Gutierrez A, Zhang L, Medberry C, et al. A whole organ regenerative medicine approach for liver replacement. Tissue Eng Part C Methods, 2011, 17(6): 677-686.
11. Uygun BE, Soto-Gutierrez A, Yagi H, et al. Organ reengineering through development of a transplantable recellularized liver graft using decellularized liver matrix. Nat Med, 2010, 16(7): 814-820.
12. Petersen TH, Calle EA, Zhao L, et al. Tissue-engineered lungs for in vivo implantation. Science, 2010, 329(5991): 538-541.
13. Karabekmez FE, Duymaz A, Moran SL. Early clinical outcomes with the use of decellularized nerve allograft for repair of sensory defects within the hand. Hand (N Y), 2009, 4(3): 245-249.
14. Vindigni V, Mazzoleni F, Rossini K, et al. Reconstruction of ablated rat rectus abdominis by muscle regeneration. Plast Reconstr Surg, 2004, 114(6): 1509-1515.
15. Conconi MT, De Coppi P, Bellini S, et al. Homologous muscle acellular matrix seeded with autologous myoblasts as a tissue-engineering approach to abdominal wall-defect repair. Biomaterials, 2005, 26(15): 2567-2574.
16. 薛辉, 陈东, 张秀英, 等. 化学去细胞肌肉组织工程支架与大鼠脊髓的生物相容性. 吉林大学学报: 医学版, 2009, 35(5): 801-804.
17. Zhang XY, Xue H, Liu JM, et al. Chemically extracted acellular muscle: A new potential scaffold for spinal cord injury repair. J Biomed Mater Res A, 2012, 100(3): 578-587.
18. Wang B, Borazjani A, Tahai M, et al. Fabrication of cardiac patch with decellularized porcine myocardial scaffold and bone marrow mononuclear cells. J Biomed Mater Res A, 2010, 94(4): 1100-1110.
19. Flynn L, Semple JL, Woodhouse KA. Decellularized placental matrices for adipose tissue engineering. J Biomed Mater Res A, 2006, 79(2): 359-369.
20. Flynn L, Woodhouse KA. Adipose tissue engineering with cells in engineered matrices. Organogenesis, 2008, 4(4): 228-235.
21. Gomillion CT, Burg KJ. Stem cells and adipose tissue engineering. Biomaterials, 2006, 27(36): 6052-6063.
22. Cherubino M, Marra KG. Adipose-derived stem cells for soft tissue reconstruction. Regen Med, 2009, 4(1): 109-117.
23. Huss FR, Kratz G. Adipose tissue processed for lipoinjection shows increased cellular survival in vitro when tissue engineering principles are applied. Scand J Plast Reconstr Surg Hand Surg, 2002, 36(3): 166-171.
24. Beahm EK, Walton RL, Patrick CW Jr. Progress in adipose tissue construct development. Clin Plast Surg, 2003, 30(4): 547-558, viii.
25. Patrick CW Jr. Tissue engineering strategies for adipose tissue repair. Anat Rec, 2001, 263(4): 361-366.
26. Flynn LE. The use of decellularized adipose tissue to provide an inductive microenvironment for the adipogenic differentiation of human adipose-derived stem cells. Biomaterials, 2010, 31(17): 4715-4724.
27. Crapo PM, Gilbert TW, Badylak SF. An overview of tissue and whole organ decellularization processes. Biomaterials, 2011, 32(12): 3233-3243.
28. 卿泉, 秦廷武. 大鼠骨骼肌脱细胞处理方法的优化研究. 中国修复重建外科杂志, 2009, 23(7): 836-839.
29. Ciapetti G, Cenni E, Pratelli L, et al. In vitro evaluation of cell/biomaterial interaction by MTT assay. Biomaterials, 1993, 14(5): 359-364.
30. 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.
31. Zuk PA, Zhu M, Ashjian P, et al. Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell, 2002, 13(12): 4279-4295.
32. Gimble JM, Guilak F. Adipose-derived adult stem cells: isolation, characterization, and differentiation potential. Cytotherapy, 2003, 5(5): 362-369.
33. Gimble JM, Katz AJ, Bunnell BA. Adipose-derived stem cells for regenerative medicine. Circ Res, 2007, 100(9): 1249-1260.
34. Dobson DE, Kambe A, Block E, et al. 1-Butyryl-glycerol: a novel angiogenesis factor secreted by differentiating adipocytes. Cell, 1990, 61(2): 223-230.
35. Rodriguez AM, Pisani D, Dechesne CA, et al. Transplantation of a multipotent cell population from human adipose tissue induces dystrophin expression in the immunocompetent mdx mouse. J Exp Med, 2005, 201(9): 1397-1405.
36. McIntosh K, Zvonic S, Garrett S, et al. The immunogenicity of human adipose-derived cells: temporal changes in vitro. Stem Cells, 2006, 24(5): 1246-1253.

Chinese Journal of Reparative and Reconstructive Surgery

PREPARATION AND BIOCOMPATIBILITY OF PORCINE SKELETAL MUSCLE ACELLULAR MATRIX FOR ADIPOSE TISSUE ENGINEERING

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