Chemical extraction method versus detergent-enzymatic method for the preparation of tissue-engineered trachea matrix

doi:10.3969/j.issn.2095-4344.2016.51.006

Abstract

Abstract:

BACKGROUND: Acellular tracheal matrix is similar to the native trachea with structure and biological performance preserved after decellularization, and it is an important aim in tissue engineering to find an effective method of decellularization.
OBJECTIVE: To select the optimal decellularization method through comparing chemical extraction method with detergent-enzymatic method for preparing rabbit tissue engineering trachea matrix.
METHODS: Thirty tracheas from New Zealand white rabbits were randomly divide into three groups. Twenty of rabbit tracheas were subjected to decellularization using 2% TritonX-100 combined with deoxyribonuclease I and ribonuclease (chemical extraction method group), and sodium deoxycholate combined with deoxyribonuclease I (detergent-enzymatic group), respectively. The other ten were given no intervention as controls. Samples were collected and observed by hematoxylin-eosin staining, Masson-trichrome staining, safranin O staining, DAPI staining and scanning electronic microscope.
RESULTS AND CONCLUSION: Hematoxylin-eosin staining demonstrated that compared with the control group, almost all cellular components of the mucosal epithelium were removed in the detergent-enzymatic and chemical extraction groups, and there were few remnant chondrocytes. Masson-trichrome staining indicated that compared with the control group, components of the mucosal layer chondrocytes in the chemical extraction and detergent-enzymatic groups were completely removed, with only part of remained chondrocytes in the cartilage zone. Glycosaminoglycan was slightly decreased both in the chemical extraction and detergent-enzymatic groups shown by Safranin O staining, but more reduction was found in the chemical extraction group. DAPI staining reveled that only a small amount of cartilage cells remained in the dense layer of cartilage and lacuna both in this two methods. Scanning electronic microscope showed that using the detergent-enzymatic method there were the hierarchical structures of native trachea, but slight disruption using the chemical extraction method. In conclusion, decellularized rabbit trachea matrix obtained by detergent-enzymatic method is better, with little disruption to the matrix.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Extracellular Matrix, Trachea, Tissue Engineering

CLC Number:

R318

Jiang Yuan, Xu Yan-fei, Zhang Si-quan, Shi Hong-can. Chemical extraction method versus detergent-enzymatic method for the preparation of tissue-engineered trachea matrix[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(51): 7642-7647.

Figures/Tables 1

References

[1] Sukumaran R,Nair RA,Jacob PM,et al.Extramedullary Plasmacytoma of the Trachea.Head Neck Pathol. 2014; 8(2):220-224.

[2] Angelelini GD,Penny WJ,el-Ghamary F,et al.The incidence and significance of early pericardial effusion after open heart surgery.Eur J Cardiothorac Surg. 1978; 1:165-168.

[3] Kojima K,Vacanti CA.Tissue engineering in the trachea.Anat Rec (Hoboken). 2014;297(1):44-50.

[4] Ott LM,Weatherly RA,Detamore MS.Overview of tracheal tissue engineering :Clinical need drives the laboratory approach.Ann Biomed Eng. 2011,39:2091-2113.

[5] Gonfiotti A,Jaus MO,Barale D,et al.The first tissue-engineered airway transplantation: 5-year follow-up results.Lancet.2014;383(9913):238-244.

[6] Haykal S,Salna M,Waddell TK,et al.dvances in Tracheal Reconstruction.Plast Reconstr Surg Glob Open.2014;2(7):e178.

[7] Góra A,Pliszka D,Mukherjee S,Ramakrishna S.Tubular Tissues and Organs of Human Body--Challenges in Regenerative Medicine.J Nanosci Nanotechnol. 2016; 16(1):19-39.

[8] Langer R,Vacanti JP.Tissue engineering. Science, 1993;260: 920-926.

[9] Roosens A,Somers P,De Somer F,et al.Impact of Detergent-Based Decellularization Methods on Porcine Tissues for Heart Valve Engineering.Ann Biomed Eng. 2016;44(9):2827-2839.

[10] Dong J,Mo X,Li Y,et al.Recent research progress of decellularization of native tissues.Sheng Wu Yi Xue Gong Cheng Xue Za Zhi.2012;29(5):1007-1013.

[11] 章方彪,史宏灿,张卫东,等.兔脱细胞同种异体气管基质衍生材料的形态特征及其生物力学分析[J].中华实验外科杂志,2014,31(3):573-576.

[12] Sun F,Pan S,Shi HC,et al.Structural integrity, immunogenicity and biomechanical evaluation of rabbit decelluarized tracheal matrix.J Biomed Mater Res A.2015;103(4):1509-1519.

[13] Yang H,Tan Q,Zhao H.Progress in various crosslinking modification for acellular matrix.Chin Med J (Engl). 2014; 127(17):3156-3164.

[14] Tomizawa Y.Late spontaneous nonanastomotic transgraft hemorrhage from biological material-impregnated fabric vascular graft may be due to autologous tissue detachment: a clinical hypothesis. Artif Organs.2014;38(12):1058-1060.

[15] Pimenta FR,Ferreira MD,Gir E,et al.Care and specialized clinical follow-up of nursing professionals who have been victims of accidents with biological material. Rev Esc Enferm USP.2013;47(1):198-204.

[16] Crapo PM,Gilbert TW,Badylak SF.An overview of tissue and whole organ decellularization processes.Biomaterials.2011;32(12):3233-3243.

[17] Rice RD,Ayubi FS,Shaub ZJ,et al.Comparison of Surgisis, AlloDerm, and Vicryl Woven Mesh grafts for abdominal wall defect repair in an animal model.Aesthetic Plast Surg. 2010;34(3):290-296.

[18] Scarritt ME,Pashos NC,Bunnell BA.A review of cellularization strategies for tissue engineering of whole organs.Front Bioeng Biotechnol.2015;3:43.

[19] Faulk DM,Johnson SA,Zhang L,et al.Role of the extracellular matrix in whole organ engineering.J Cell Physiol.2014;229(8):984-989.

[20] Zhou J,Fritze O,Schleicher M,et al.Impact of heart valve decellularization on 3-D ultrastructure, immunogenicity and thrombogenicity. Biomaterials. 010;31(9): 2549-2554.

[21] Annunziata C,Alessandra C,Attila T,et al.Inflammation in tissue engineering: the Janus between engraftment and rejection.Eur J Immunol.2015;45(12):3222-3236.

[22] Keane TJ,Swinehart IT,Badylak SF.Methods of tissue decellularization used for preparation of biologic scaffolds and in vivo relevance.Methods.2015;84:25-34.

[23] Crapo PM,Gilbert TW,Badylak SF.An overview of tissue and whole organ decellularization processes. Biomaterials.2011;32(12):3233-3243.

[24] Maghsoudlou P,Totonelli G,Loukogeorgakis SP,et al.A decellularization methodology for the production of a natural acellular intestinal matrix.J Vis Exp. 2013;(80): e50658.

[25] Perea-Gil I,Uriarte JJ,Prat-Vidal C,et al.In vitro comparative study of two decellularization protocols in search of an optimal myocardial scaffold for recellularization.Am J Transl Res.2015;7(3):558-573.

[26] Shupe T,Williams M,Brown A,et al.Method for the decellularization of intact rat liver.Organogenesis. 2010;6(2):134-136.

[27] Cebotari S,Tudorache I,Jaekel T,et al.Detergent decellularization of heart valves for tissue engineering: toxicological effects of residual detergents on human endothelial cells.Artif Organs.2010;34(3):206-210.

[28] Xu H,Xu B,Yang Q,et al.Comparison of decellularization protocols for preparing a decellularized porcine annulus fibrosus scaffold.PLoS One.2014;9(1):e86723.

[29] Badylak SF.The extracellular matrix as a biologic scaffold material. Biomaterials. 2007;28:3587-3593.

[30] Uriel S,Labay E,Francis-Sedlak M,et al.Extraction and assembly of tissue-derived gels for cell culture and tissue engineering.Tissue Eng Part C Methods. 2009; 15:309-321.

[31] Farag A,Vaquette C,Theodoropoulos C,et al.Decellularized periodontal ligament cell sheets with recellularization potential.J Dent Res. 2014;93(12):1313-1319.

[32] Jungebluth P,Go T,Asnaghi A,et al.Structural and morphologic evaluation of a novel detergent-enzymatic tissue-engineered tracheal tubular matrix.J Thorac Cardiovasc Surg. 2009;138(3):586-593.

[33] Illien-Jünger S,Sedaghatpour DD,Laudier DM,et al.Development of a bovine decellularized extracellular matrix-biomaterial for nucleus pulposus regeneration.J Orthop Res.2016;34(5):876-888.

[34] Yu BT,Li WT,Song BQ,et al.Comparative study of the Triton X-100-sodium deoxycholate method and detergent-enzymatic digestion method for decellularization of porcine aortic valves.Eur Rev Med Pharmacol Sci.2013;17(16):2179-2184.