簡易檢索 / 詳目顯示

研究生: 陳松青
Sung-Ching Chen
論文名稱: 多孔性去細胞牛心包膜於心肌組織再生上之研究
Porous Acellular Bovine Pericardia for Myocardial Tissue Regeneration
指導教授: 宋信文
Hsing-Wen Sung
口試委員:
學位類別: 博士
Doctor
系所名稱: 工學院 - 化學工程學系
Department of Chemical Engineering
論文出版年: 2005
畢業學年度: 93
語文別: 英文
論文頁數: 81
中文關鍵詞: 心肌梗塞人工補綴片去細胞牛心包膜間葉幹細胞心肌組織再生組織工程
外文關鍵詞: myocardial infarction, artificial patch, acellular bovine pericardium, mesenchymal stem cell, myocardial tissue regeneration, tissue engineering
相關次數: 點閱:2下載:0
分享至:
查詢本校圖書館目錄 查詢臺灣博碩士論文知識加值系統 勘誤回報
  • Clinically, adult human myocardium lacks the possibility of regeneration after myocardial infarction. This results in a progressive loss of functional myocardium and a successive reduction in cardiac performance. Non-living synthetic materials have been widely used to repair myocardial defects; however, material-related failures do occur. In the study, an acellular bovine pericardium with a porous structure fixed by genipin (the AGP patch) was prepared and employed to repair a surgically created myocardial defect in the right ventricle of a rat model. A commercially available expanded polytetrafluoroethylene (e-PTFE) patch was used as a control. At retrieval, a computerized mapping system was employed to acquire local epicardial electrograms of each implanted sample and the appearance of each retrieved sample was grossly examined. The retrieved samples were then processed for histological examinations.
    The amplitude of local electrograms on the AGP patch increased significantly with increasing the implantation duration, while only low-amplitude electrograms were observed on the e-PTFE patch throughout the entire course of the study. No aneurysmal dilation of the implanted patches was seen for both studied groups. Additionally, no tissue adhesion was observed on the outer (epicardial) surface of the AGP patch, while a moderate tissue adhesion was observed on the e-PTFE patch. On the inner (endocardial) surface, intimal thickening was observed for both studied groups; however, no thrombus formation was found. Intact layers of endothelial and mesothelial cells were identified on the inner and outer surfaces of the AGP patch, respectively.
    At 4-week postoperatively, smooth muscle cells together with neo-muscle fibers (with a few neo-collagen fibrils), neo-glycosaminoglycans, and neo-capillaries were observed to fill the pores in the AGP patch, an indication of tissue regeneration. These observations were more pronounced at 12-week postoperatively. In contrast, no apparent tissue regeneration was observed in the e-PTFE patch. In summary, the AGP patch may preserve the structure of the right ventricle and prevent aneurysmal dilation while providing the potential for tissue regeneration. These results indicated that the AGP patch holds promise to become a suitable patch for surgical repair of myocardial defects. However, cardiomyocytes were not found within the AGP patch, a limitation of the study.
    To overcome this problem, syngenic rat bone-marrow derived mesenchymal stem cells (MSCs) were seeded onto the AGP patch (the MSC-seeded AGP patch) and implanted in the same animal model. After seeding MSCs onto the AGP patch, the cells had an uniform and viable fibroblast-like morphology and revealed a good interconnectivity. At retrieval, all the gross and histological observations were similar to those seen in the AGP patch, with the exception of the following phenomenon. Cardiomyocytes, stained positively by troponin T and myosin heavy chain, were observed in the MSC-seeded AGP patch, while the AGP patch was negatively stained. Additionally, MSCs induced angiogenesis in the MSC-seeded AGP patch.
    In conclusion, cardiomyocytes together with neo-muscle fibers were observed in the MSC-seeded AGP patch, an indication of myocardial tissue regeneration. The MSC-seeded AGP patch may permit the construction of myocardial defects.


    Table of Content ABSTRACT…...………………………………………………………..I TABLE OF CONTENT……………………………………………….III LIST OF FIGURES…….………………………………………..……VI Chapter 1. Introduction 1 Chapter 2. Tissue Regeneration Observed in a Porous Acellular Bovine Pericardium Used to Repair a Myocardial Defect in the Right Ventricle of a Rat Model 2.1 Materials and Methods 5 2.1.1 Preparation of Test Samples 5 2.1.2 Animal Study 6 2.1.3 Computerized Mappings (Epicardial Electrograms) 8 2.1.4 Histological Examinations 8 2.2 Results 9 2.2.1 Test Samples 9 2.2.2 Computerized Mappings (Epicardial Electrograms) 12 2.2.3 Gross Examination 12 2.2.4 Histological Findings 17 2.3 Discussion 24 2.4 Conclusions 29 Chapter 3. Mesenchymal Stem Cell-seeded Porous Acellular Bovine Pericardia as a Patch to Repair a Myocardial Defect in a Syngenic Rat Model 3.1 Materials and Methods 30 3.1.1 Experimental Animals 30 3.1.2 Preparation of the Porous Acellular Bovine Pericardium 31 3.1.3 Isolation and Culture of Bone Marrow-derived Mesenchymal Stem Cells (Bone marrow MSCs) 32 3.1.4 Construction of the Bone Marrow MSC-seeded Patch (In Vitro Graft Preparation, Cell Seeding onto the CMAGP Patches) 33 3.1.5 Animal Study 34 3.1.6 Echocardiography (Assessment of Cardiac Function) 35 3.1.7 Histological Examinations 36 3.1.8 Statistical Analysis 38 3.2 Results 38 3.2.1 Preparation of the Porous Acellular Bovine Pericardium 38 3.2.2 In Vitro Evaluation of Bone Marrow MSC-seeded Patch (the BMMSC Patch) 40 3.2.3 Echocardiography (Assessment of Cardiac Function) 43 3.2.4 Gross Examination 43 3.2.5 Histological Findings 44 3.3 Discussion 57 3.4 Conclusions 63 References 65 著作目錄 75 作者簡歷 81

    References
    1. Pasumarthi KB, Field LJ. Cardiomyocyte cell cycle regulation. Circ Res. 2002;90:1044-1054.
    2. Jugdutt BI. Prevention of ventricular remodelling post myocardial infarction: timing and duration of therapy. Can J Cardiol. 1993;9:103-114.
    3. Takemura G, Ohno M, Hayakawa Y, Misao J, Kanoh M, Ohno A, Uno Y, Minatoguchi S, Fujiwara T, Fujiwara H. Role of apoptosis in the disappearance of infiltrated and proliferated interstitial cells after myocardial infraction. Circ Res. 1998;82:1130-1138.
    4. Akhyari P, Fedak PWM, Weisel RD, Lee TYJ, Verma S, Mickle DAG, et al. Mechanical stretch regimen enhances the formation of bioengineered autologous cardiac muscle grafts. Circulation. 2002;106(suppl I):I-137-142.
    5. Ozawa T, Mickle DAG, Weisel RD, Koyama N, Wong H, Ozawa S, et al. Histologic changes of nonbiodegradable and biodegradable biomaterials used to repair right ventricular heart defects in rats. J Thorac Cardiovasc Surg. 2002;124:1157-1164.
    6. Oechslin EN, Harrison DA, Harris L. Reoperation in adults with repair of tetralogy of Fallot: indications and outcomes. J Thorac Cardiovasc Surg. 1999;118:245-251.

    7. Askari AT, Unzek S, Popovic ZB, Goldman CK, Forudi F, Kiedrowski M, et al. Effects of stromal-cell-derived factor 1 on stem-cell horming and tissue regeneration in ischaemic cardiomyopathy. Lancet. 2003;362:697-703.
    8. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR. Multilineage potential of adult human mesenchymal stem cells. Science. 1999;284:143-147.
    9. Minguell JJ, Erices A, Conget P. Mesenchymal stem cells. Exp Biol Med. 2001;226:507-520.
    10. Devine SM. Mesenchymal stem cells: will they have a role in the clinic? J Cell Biochem Suppl. 2002;38:73-79.
    11. Xu W, Zhang X, Qian H, Zhu W, Sun X, Hu J, Zhou H, Chen Y. Mesenchymal stem cells from adult human bone marrow differentiate into a cardiomyocyte phenotype in vitro. Exp Biol Med. 2004;229:623-631.
    12. Orlic D, Kajstura J, Chimenti S, Jakoniuk I, Anderson SM, Li B, Pickel J, McKay R, Nadal-Ginard B, Bodine DM, Leri A, Anversa P. Bone marrow cells regenerate infracted myocardium. Nature. 2001;410:701-705.
    13. Toma C, Pittenger MF, Cahill KS, Byrne BJ, Kessler PD. Human mesenchymal stem cells differentiate to a cardiomyocyte phenotype in the adult murine heart. Circulation. 2002;105:93-98.
    14. Tomita S, Mickle DA, Weisel RD, Jia ZQ, Tumiati LC, Allidina Y, Liu P, Li RK. Improved heart function with myogenesis and angiogenesis after autologous porcine bone marrow stromal cell transplantation. J Thorac Cardiovasc Surg. 2002;123:1132-1140.
    15. Strauer BE, Brehm M, Zeus T, Köstering M, Hernandez A, Sorg RV, Kögler G, Wernet P. Repair of infracted myocardium by autologous intracoronary mononuclear bone marrow cell transplantation in humans. Circulation. 2002;106:1913-1918.
    16. Noishiki Y, Miyata T. A simple method to heparinize biological materials. J Biomed Mater Res. 1986;20:337-346.
    17. Noishiki Y, Miyata T, Kodaira K. Development of a small calibre vascular graft by a new crosslinking method incorporating slow heparin release collagen and natural tissue compliance. ASAIO J. 1986;32:114-119.
    18. Courtman DW, Pereira CA, Kashef V, MicComb D. Development of a pericardial acellular matrix biomaterial: Biochemical and mechanical effects of cell extraction. J Biomed Mater Res. 1994;28:655-666.
    19. Chang Y, Tsai CC, Liang HC, Sung HW. In vivo evaluation of cellular and acellular bovine pericardia fixed with a naturally occurring crosslinking agent (genipin). Biomaterials. 2002;23:2447-2457.
    20. Sung HW, Chen CN, Huang RN, Hsu JC, Chang WH. In vitro surface characterization of a biological patch fixed with a naturally occurring crosslinking agent. Biomaterials. 2000;21:1353-1362.
    21. Chang Y, Tsai CC, Liang HC, et al. Reconstruction of the right ventricular outflow tract with a bovine jugular vein graft fixed with a naturally occurring crosslinking agent (genipin) in a canine model. J Thorac Cardiovasc Surg. 2001;122:1208-1218.
    22. Chang Y, Lee MH, Liang HC, Hsu CK, Sung HW. Acellular bovine pericardia with distinct porous structures fixed with genipin as an extracellular matrix. Tissue Eng. 2004;10:881-892.
    23. Wei HJ, Liang HC, Lee MH, Huang YC, Chang Y, Sung HW. Construction of varying porous structures in acellular bovine pericardia as a tissue-engineering extracellular matrix. Biomaterials. 2005;26:1905-1913.
    24. Sung HW, Chang Y, Chiu CT, Chen CN, Liang HC. Crosslinking characteristics and mechanical properties of a bovine pericardium fixed with a naturally occurring crosslinking agent. J Biomed Mater Res. 1999;47:116-126.
    25. Ramshaw AM, Casagranda F, White JF, Edwards GA, Hunt JA, Williams DF, et al. Effects of mesh modification on the structure of a mandrel-grown biosynthetic vascular prosthesis. J Biomed Mater Res. 1999;47:309-315.
    26. Martin I, Shastri VP, Padera RF, Yang J, Mackay AJ, Langer R, et al. Selective differentiation of mammalian bone marrow stromal cells cultured on three-dimensional polymer foams. J Biomed Mater Res. 2001;55:229-235.
    27. Lee JM, Haberer SA, Boughner DR. The bovine pericardial xenograft. I: Effect of fixation in aldehydes without constraint on the tensile viscoelastic properties of bovine pericardium. J Biomed Mater Res. 1989;23:457-475.
    28. Ozawa T, Mickle DAG, Weisel RD, Koyama N, Ozawa S, Li RK. Optimal biomaterial for creation of autologous cardiac grafts. Circulation. 2002;106(suppl I): I-176-182
    29. Wu TJ, Doshi RN, Huang HLA, Blanche C, Kass RM, Trento A, et al. Simultaneous biatrial computerized mapping during permanent atrial fibrillation in patients with organic heart disease. J Cardiovasc Electrophysiol. 2002;13:571-577.
    30. Prophet EB, Mills B, Arrington JB, Sobin LH. Laboratory Methods in Histotechnology. 2nd ed. Washington: American Registry of Pathology; 1994, p. 136.
    31. Allaire E, Guettier C, Bruneval P, Plissonnier D, Michel JB. Cell-free arterial grafts: Morphologic characteristics of aortic isografts, allografts, and xenografts in rats. J Vasc Surg. 1994;19:446-456.
    32. Courtman DW, Errett BF, Wilson GJ, et al. The role of crosslinking in modification of the immune response elicited against xenogenic vascular acellular matrices. J Biomed Mater Res. 2001;55:576-586.
    33. Bader A, Schiling T, Teebken OE. Tissue engineering of heart valves-human endothelial cell seeding of detergent acellularized porcine valves. Euro J Cardiothorac Surg. 1998;14:279-284.
    34. Matsuda T, Nakayama Y. Surface microarchitectural design in biomedical applications: In vitro transmural endothelialization on microporous segmented polyurethane films fabricated using an excimer laser. J Biomed Mater Res. 1996;31:235-242.
    35. Izutani H, Gundry SR, Vricella LA, Xu H, Bailey LL. Right ventricular outflow tract reconstruction using a GoreTex membrane monocusp valve in infant animals. ASAIO J. 2000;46:553-555.
    36. Uemura H, Yagihara T, Kawahira Y, Yoshikawa Y. Kitamura S. Total cavopulmonary connection in children with body weight less than 10 kg. Eur J Cardiothorac Surg. 2000;17:543-549.
    37. Macchi E, Cavalieri M, Stilli D, Musso E, Baruffi S, Olivetti G, et al. High-density epicardial mapping during current injection and ventricular activation in rat hearts. Am J Physiol. 1998;275: H1886-1897.
    38. Whitaker D, Papadimitriou JM, Walters M. The mesothelium: its fibrinolytic properties. J Pathol. 1982;136:291-299.
    39. Scott SM, Barth MG, Gaddy LR, Ahl ET Jr. The role of circulating cells in the healing of vascular prostheses. J Vasc Surg. 1994;19:585-593.
    40. Chang Y, Liang HC, Wei HJ, Chu CP, Sung HW. Tissue regeneration patterns in acellular bovine pericardia implanted in a canine model as a vascular patch. J Biomed Mater Res. 2004;69A:323-333.

    41. Schürch W, Seemayer TA, Gabbiani G. Myofibroblast: In: Sternberg SS, editor. Histology for Pathologists. New York, NY: Raven Press Publishers; 1992, p. 109-144.
    42. Shinoka T, Ma PX, Shum-Tim D, Breuer CK, Cusick RA, Zund G, et al. Tissue-engineered heart valves: autologous valve leaflet replacement study in a lamb model. Circulation. 1996;94(9 Suppl): II-164-168.
    43. Yoo KJ, Li RK, Weisel RD, Mickle DAG, Li G, Yau TM. Autologous smooth muscle cell transplantation improved heart function in dilated cardiomyopathy. Ann Thorac Surg. 2000;70:859-865.
    44. Amenta PS, Amenta PS. Muscle tissue: In: Elias H, Pauly JE, editors. Histology From Normal Microanatomy To Pathology. Padova: Piccin Nuova Libraria S.p.A.; 1997, p. 153.
    45. Li RK, Jia ZQ, Weisel RD, Mickle DAG, Zhang J, Mohabeer MK, et al. Cardiomyocyte transplantation improved heart function. Ann Thorac Surg. 1996;62:654-661.
    46. Wakitani S, Saito T, Caplan AI. Myogenic cells derived from rat bone marrow mesenchymal stem cells exposed to 5-aza-cytidine. Muscle Nerve. 1995;18:1417-1426.
    47. Tomita S, Li RK, Weisel RD, Mickle DAG, Kim EJ, Sakai T, Jia ZQ. Autologous transplantation of bone marrow cells improves damaged heart function. Circulation. 1999;100[suppl]:II247-256.

    48. Krupnick AS, Kreisel D, Engels FH, Szeto WY, Plappert T, Popma SH, Flake AW, Rosengard BR. A novel small animal model of left ventricular tissue engineering. J Heart Lung Transplant. 2002;21:233-243.
    49. Miyagawa S, Sawa Y, Taketani S, Kawaguchi N, Nakamura T, Matsuura N, Matsuda H. Myocardial regeneration therapy for heart failure hepatocyte growth factor enhances the effect of cellular cardiomyoplasty. Circulation. 2002;105:2556-2561.
    50. Jones JE, Mendes L, Rudd MA, Russo G, Loscalzo J, Zhang YY. Serial noninvasive assessment of progressive pulmonary hypertension in a rat model. Am J Physiol Heart Circ Physiol. 2002;283:364-371.
    51. Li RK, Jia ZQ, Weisel RD, et al. Cardiomyocyte transplantation improves heart function. Ann Thorac Surg. 1996;62:654-660.
    52. Li RK, Jia ZQ, Weisel RD, et al. Smooth muscle cell transplantation into myocardial scar tissue improves heart function. J Mol Cell Cardiol. 1999;31:513-522.
    53. Matsubayashi K, Fedak PWM, Mickle DAG, Richard D, Weisel RD, Ozawa T, Li RK. Improved left ventricular aneurysm repair with bioengineered vascular smooth muscle grafts. Circulation. 2003;108 [suppl II]:II219-225.

    54. Chang Y, Chen SC, Wei HJ, Wu TC, Liang HC, Lai PH, Yang HH, Sung HW. Tissue regeneration observed in a porous acellular bovine pericardium used to repair a myocardial defect in the right ventricle of a rat model. J Thorac Cardiovasc Surg. (in press)
    55. Malone JM, Brendel K, Duhamel RC, Reinert RL. Detergent-extracted small-diameter vascular prostheses. J Vasc Surg. 1984;1:181-191.
    56. Graham LM, Vinter DW, Ford JW, Kahn RH, Burkel WE, Stanley JC. Endothelial cell seeding of prosthetic vascular grafts. Early experimental studies with cultured autologous canine endothelium. Arch Surg. 1980;115:929-933.
    57. Wang JS, Shum-Tim D, Galipeau J, et al. Marrow stromal cells for cellular cardiomyoplasty: feasibility and potential clinical advantages. J Thorac Cardiovasc Surg. 2000;120:999-1005.
    58. Davani S, Marandin A, Mersin N, Royer B, Kantelip B, Hervé P, Etievent JP, Kantelip JP. Mesenchymal progenitor cells differentiate into an endothelial phenotype, enhance vascular density, and improve heart function in a rat cellular cardiomyoplasty model. Circulation. 2003;108[suppl II]:II253-258.
    59. Asahara T, Murohara T, Sullivan A, Silver M, Zee RVD, Li T, Witzenbichler B, Schatteman G, Isner JM. Isolation of putative progenitor endothelial cells for angiogenesis. Science. 1997;275:964-967.
    60. Liang HC, Chen CT, Chang Y, Huang YC, Chen SC, Sung HW. A novel angiogenic agent (Ginsenoside Rg1) loaded in an acellular biological tissue for tissue regeneration. Tissue Eng. (in press)

    無法下載圖示 全文公開日期 本全文未授權公開 (校內網路)
    全文公開日期 本全文未授權公開 (校外網路)

    QR CODE