跳到主要內容

臺灣博碩士論文加值系統

(18.97.14.87) 您好!臺灣時間:2025/03/18 11:57
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果 :::

詳目顯示

我願授權國圖
: 
twitterline
研究生:蔡曉雯
研究生(外文):Shesu-Wen Tsai
論文名稱:含褐藻酸鹽之膠原蛋白基材之製備與質傳模擬
論文名稱(外文):Preparation and Simulation of Mass Transfer for Alginate - Containing Collagen Matrix
指導教授:王盈錦蔡瑞瑩
指導教授(外文):Yng-Jiin WangRuey-Yug Tray
學位類別:博士
校院名稱:國立陽明大學
系所名稱:醫學工程研究所
學門:工程學門
學類:生醫工程學類
論文種類:學術論文
論文出版年:1999
畢業學年度:87
語文別:中文
中文關鍵詞:膠原蛋白褐藻酸鹽質傳模擬
外文關鍵詞:collagenalginatemass transfer simulation
相關次數:
  • 被引用被引用:0
  • 點閱點閱:237
  • 評分評分:
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:0
本研究主要在於利用褐藻酸鹽微膠囊技術,製備一含膠原蛋白纖維的細胞培養顆粒擔體。並以理論評估生長因子於膠原基質上的濃度分布。膠原蛋白是結締組織中主要的構造性蛋白質,經適當的處理可具高度生物相容性,及低免疫排斥性。高純度之膠原蛋白在37℃、pH 7.4時可重組形成三度空間的網狀結構。目前商品化的膠原顆粒製備方法,乃將動物膠原纖維懸浮液直接滴入液態氮中,以急速冷凍製成,再以戊二醛固定。此方法所製成之顆粒因易含有雜質,生物相容性不佳而易引發免疫反應,因此無法直接作醫療應用。本研究乃利用褐藻酸鹽凝膠特性,將褐藻酸鹽與膠原蛋白纖維混合液,於4℃下滴入鈣離子溶液中,以製得含膠原蛋白纖維的顆粒擔體。一但將褐藻酸鹽膠體以檸檬酸液化後,在37℃下膠原蛋白即可在顆粒內重組。相較於傳統的微膠囊系統,GH3或L929細胞在此含膠原蛋白的擔體內均有一較良好生長表態。若能於擔體內加入適當的生長因子則應更能促進細胞的生長情形。本研究之理論模擬主要在於修正過去文獻中,有關複合基材上藥物釋放理論中”perfect sink”的邊界條件。以基材邊界”matching”作理論推導,藉以評估生長因子於培養基材上的濃度分布情形,以作為設計細胞培養基材的依據。
The aim of this study is to prepare a collagen-based microcapsule for cell culture using alginate as an encapsulation medium. Furthermore, theoretical modeling has been conduct to simulate the concentration distribution of growth factor within the cell culture matrix. Collagen is the major structural protein in the extracellular matrices of connective tissues. With proper treatment, its immune response can be eliminated and provide a better biocompatibility. Purified collagen molecules can be reconstituted into a three-dimensional fibrous network at 37℃, pH = 7.4. Present commercialized products of collagen beads are obtained by quenched in liquid nitrogen and then crosslinked with glutaraldehyde. Such kind of gel beads can''t be applied as a biomaterial directly because of its poor biocompatibility. We introduce a method of preparing microcapsules containing collagen fibrous network. This method takes the advantage of the gelation properties of alginate and forms spherical gel beads by dropping a mixture of collagen and alginate into a solution of CaCl2 at 4℃. Collagen was then reconstitute within the microcapsules at 37℃ after alginate was liquefied with citrate. Both of the growth oh the GH3 rat pituitary tumor cell and the L929 fibroblastic-like cell were studied for the present microcapsules. The results indicate that both cells proliferate faster in the collagen-containing capsule than those of in the conventional collagen-free microcapsules. It is believed that by applying specific growth factor, the cell proliferation rate can be promoted. In this study, a mass transfer model is developed to simulate the concentration distribution of growth factor molecules. The established model can be applied in designing a cell culture material.
封面
目錄
中文摘要
ABSTRACT
第一章 緒論
1.1. 細胞培養擔體發展之回顧
1.2. 膠原蛋白簡介
(1)膠原蛋白的分子結構
(2)膠原蛋白的純化與重組
(3)膠原蛋白的分類及生醫應用
1.3. 褐藻酸鹽簡介
(1)褐藻酸鹽的分子結構、物性、化性
(2)褐藻酸鹽的生醫應用
1. 4. TGF-B(Transformation Growth Factor-)
1. 5. BMPs(Bone Morphogenetic Proteins)
1. 6 理論質傳模型
(1)膨潤與溶蝕型之藥物釋放動力學
(2)滲透型之釋放動力學
1.7 研究動機與目的
第二章 理論模式的建立
2.1. 質傳理論模擬
2.2. 生長因子消耗速率常數分析
第三章 實驗儀器與藥品
儀器:
藥品:
第四章
褐藻酸鹽/膠原纖維微粒之製備與細胞培養實驗方法
4.1. 動物膠原蛋白的萃取與純化
4.2. 動物膠原蛋白的純度鑑定與濃度測試
(1)膠原蛋白純度測定
(2)膠原蛋白濃度測定
4.3. 含褐藻酸鹽之膠原基材熱性質分析
4.4. 含褐藻酸鹽之膠原基材凝膠性質測試
4.5. 含褐藻酸鹽之膠原微粒之製備
4.6. 含褐藻酸鹽之膠原微粒機械性質測試
4.7. 含褐藻酸鹽之膠原微粒包覆3T3或L929細胞的製備與培養
4.8. 含褐藻酸鹽之膠原微粒包覆GH3細胞的製備與培養
第五章 褐藻酸鹽/膠原纖維微粒製備與細胞培養 結果與討論
5. 1. 動物膠原蛋白的萃取與純化
(1)純度測定
(2)濃度量測
(3)掃描式顯維鏡維結構之觀察
5.2. 含褐藻酸鹽之膠原蛋白基材之熱性質分析
5.3. 含褐藻酸鹽之膠原蛋白基材凝膠性質測試
5.4. 褐藻酸鹽/膠原蛋白顆粒經serum - free之medium清洗不同時間後之電顯圖
表17. 動物膠原/褐藻酸鹽不同比例之混合
5.5. 含褐藻酸鹽之膠原蛋白顆粒電顯圖
5.6. 褐藻酸鹽/膠原蛋白顆粒機械性質測試
5.7. 褐藻酸鹽/膠原蛋白顆粒包覆細胞培養之生長曲線
5.8. 褐藻酸鹽/膠原蛋白微粒包覆GH3細胞之生長曲線
第六章 理論質傳模擬參數建立實驗方法
6. 1. 散係數的求取
6. 2. BMPs(Bone morphogenetic proteins)的萃取與純化
6. 3. 生長因子(TGF-B、BMPs)對細胞生長之動力影響
第七章 理論質傳模擬參數之建立
7.1. 擴散係數的求取
7.2. BMPs的萃取與純化
7.4. TGF-B對細胞生長動力之影響
第八章 結論
APPENDIX A
APPENDIX B
參考文獻
Hellman, KB., Picciolo, GL. and Fox,.CF. (1994).“ Prospects for Application of Biotechnology-Derived Biomaterials,” Journal of Cellular Biochemistry, 56 : 210-224
Lacy, PE., Hegre, OD., Gerasimidi-Vazeou, A., Gentile, FT. and Dionne, KE.(1991).” Maintenance of normoglycemia in diabetic mice by subcutaneous xenografts of encapsulated islets”. Science. 254 : 1782-1784,
Lanza, RP., Borland, KM., Staruk, JE., Appel, MC., Solomon, BA. and Chick WL.(1992) ”Transplantation of encapsulated canine islets into spontaneously diabetic BB/Wor rats without immunosuppression”. Endocrinology. 131:637-642
Cieslinski, DA., Humes, HD.(1994).”Tissue engineering of a bioartificial kidney”. Biotech. Bioeng. 43 : 678-681
Fu, XW. and Sun, AM.(1989).”Microencapsulated parathyroid cells as a bioartificial parathyroid. In vivo studies,” Transplantation. 47 : 432-435,
Nilsson, K., Buzsaky, F. and Mosbachy, K. (1984) “Growth of anchorage-dependent cells on macroporous microcarriers,” Bio/Technology, 4 : 989-990
Altankov, G. et al, (1991) “Synthesis of protein —coated gelatin microsphere and their use as microcarrier for cell culture,” J. Biomater. Sci., Polym. Edn., 2 : 81-89
Karel, SF., Libicki, SB. and Roberston, CR. (1985) “The immobilization of whole cells :Engineering Principle,” Chem. Eng. Sci. 40: 1321-1354
Panikkar, B., Hari, PR. and Sharma CP.”(1997).”Modified polyacrylamide microspheres as immunosorbent.” Art.Cells,Blood Sub. & Immob.Biotech.. 25:541-550,
Kiremitzi, M. and Piskin, E. (1990) “ Cell adhesion on the surfaces of polymeric beads,” Biomat., Art. Cell, Art. Org. 18 : 599-603
Zekorn, T., Siebers, U., Horcher, A., Schnettler, R., Klock, G., Bretzel, RG., Zimmermann, U.and Federlin, K.(1992).”Barium-alginate beads for immunoisolated transplantation of islets of Langerhans.” Transp. Proc. 24 : 937-9,
Gaserod O., Smidsrod O. and Skjak-Braek G.(1998).“Microcapsules of alginate-chitosan--I. A quantitative study of the interaction between alginate and chitosan.” Biomaterials. 19:1815-1825,
Lin, F. and Sun, AM.. (1980)”Microencapsulated islets as bioartificial endocrine pancreas,” Science ,210 : 908-910
Wong, H., Sc, B.. and Chang,.TMS (1991) “ The microencapsulation of cells within alginate poly-l-lysine microcapsules prepared with the standard single step drop technique : histologically identified membrane imperfections and the associated graft rejection,” Biomat. Art. Cell & Immob. Biotech. ,19 :675-686
Grandolfo, M., D’Andrea, P., Paoletti, S., Martina, M., Silvestrini, G., Bonucci, E. and Vittur, F. (1993) “Culture and differentiation of condrocytes entrapped in alginate gels,” Calcif. Tissue Int. 52:42-48
Sabelman, EE. (1985). “ Biology, Biotechnology and Biocompatibility of Collagen,” In Williams, DF. (ed.) Biocompatibility of Tissue Analogs: CRC Press.
Nimni, ME. et al. (1988). “ Bioprosthesis Derived from Cross-Linked and Chemically Modified Collagenous Tissue,” In Nimni,.M. E. (ed.) Collagen, Vol. 3.
Lynch, MP., Stein, JL. Stein, GS. and Lian, JB. (1995). “The Influence of Type 1 Collagen on the Develo[ment and Mqintenance of the Osteoblasts Phenotype in Primary and Passaged Rat Calvarial Osteoblasts: Modification of Expression of Genes Supporting Cell Growth, Adhesion, and Extracellular Matrix Mineralization, ”Experimental Cell Research, 216 : 35-45.
Susante, JLC. et al.(1995). “Culture of chondrocytes in alginate and Collagen Carrier Gels,” Acta Orthop Scand,.66 : 549-556.
Alberts, B., Bray, D., Lewis, J.,. Raff, M., Roberts, K. and Watson, JD.. (1994). Molecular Biology of The Cell. New York and London: Garland Publishing, Inc.: 978-985.
Bronzino, JD. (1995). Biomedical Engineering Handbook, Press:Chap 45
Timpl, R. (1982). “Antibodies to collagens and procollagens,” Methods in Enzymology, Vol. 82
Huang-Lee LL. and Nimni, ME. (1993).”Fibroblast contraction of collagen matrices with and without covalently bound hyaluronan.”J. of Biomat. Sci., Poly. Edi.5:99-109
Karube, I. and Nishida, T.( 1979) “Effect of alcohols on the collagen-phosphatidylcholine interaction." Biochimica et Biophysica Acta. 581:106-113,
Hsu, CY.and Tsay, R..(1997).”A study on the microstructure and permeation properties of collagen matrix,” Institute of Biomedical Engineering, National Yang-Ming University, Taiwan, Master Thesis.
Painter, TJ. ”algal polysaccharides” p263-285
Clark, AH. and Ross-Murphy, SB. “Structure and Mechanical Properties of Biopolymer Gels” p 125-129
Smidsrod, O. (1973).”Molecular Basis for Physical Properties of Alginates in the Gel State,”
Morris, ER., Rees, DA., Thom, D. and Boyd, J.(1978).”Chiroptical and stoichiometric evidence of a specific primary dimerisation process in alginate gelation,” Carbohydrate Research 66 : 145-154
Martinsen A., Skjak, BG. Smidsrod, O. (1989)”Alginate as immobilization material I. Correlation between chemical and physical properties of alginate gel beads. Biotechnol. Bioeng.33 : 79-89.
Thu, B., Bruheim, P., Espevik, T., .Smidsrod, O., Soon-Shiong, P. and Skjak-Brak G.( 1996).”Alginate polycation microcapsules,” Biomaterials, 17 : 1031-1040
Thu, BP., Bruheim, T., Espevik., O., Smidsrod, Soon-Shiong, P. and Skjak-Brak, G. (1996).”Alginate polycation microcapsules,” Biomaterials, 17 : 1069-1079
Smidsrod, O. and Skjak-Brek, G. (1990). “Alginate as Immobilization Matrix for Cells,” Trends in Biotech, 8 : 71-78
Sinacore, MS., Bernard, CC. and Robert, B. (1989).”Entrapment and Growth of Murine Hybridoma Cells in Calcium Alginate Gel Microbeads,” Bio/Technology, 7 :1275-1279
Cheetham, PSJ., Garrett, C. and Clark, J. (1985) “Isomaltulose production using immobilized cells,”Biotechnol. Bioeng. 27: 471-481.
Tanaka, H., Matsumura, M. and Veliky, IA. (1984).”Diffusion characteristics of substrates in Ca-alginate gel beads,” Biotechnol. Bioeng.16 : 53-58
Langer, R. and Vacanti, .JP. (1993). Science, Vol. 260: 920-926.
Harris, SE., Bonewald, LF., Harris, MA,. Sabatini,M., Dallas,S., Feng, JQ. Ghosh-Choudhury,N., Wozney, J. and Mundy, GR.. (1994). “ Effects of Transforming Growth Factor  on Bone Nodule Formation and Expression of Bone Morphogenetic Protein 2, Osteocalcin, Osteopontin, Alkaline Phosphatase, and Type I Collagen mRNA in Long-Term Cultures of Fetal Rat Calvarial Osteoblasts,” Journal of Bone and Mineral Research, 9: 855-863.
Talley, DJ., Lajiness, E. and Nagodawithana, K.. (1995). “ Transforming Growth Factor-Beta Inhibition of Mineralization by Neonatal Rat Osteoblasts in Monolayer and Collagen Gel Culture,” Vitro Cell. Dev. Biol., 31: 274-282.
Komaki, M., Katagiri,T. and Suda, T.. (1996). “ Bone Morphogenetic Protein-2 Does Not Alter The Differentiation Pathway of Committed Progenitors of Osteoblasts and Chondroblasts,” Cell & Tissue Research, 284: 9-17.
Yamaguchi, A., Ishizuya,T., Kintou,N., Wada,Y., Katagiri,T., Wozney, JM., Rosen, V. and Yoshiki, S. (1996). “ Effects of BMP-2, BMP-4, and BMP-6 on Osteoblastic Differentiation of Bone Marrow-Derived Stromal Cell Lines, ST2 and MC3T3-G2/PA6,” Biochemical and Biophysical Research Communications, 220: 336-371.
Aldinger, G., Herr,G., Kiisswetter, W., Reis,HJ., Thielemann, FW. and Holz, U.. (1991). “ Bone Morphogenetic Protein: A Review,” International Orthopaedics (SICOT), 15: 169-177.
Urist, MR., Lietze, A. and Dawson, E. (1984). “ -Tricalcium Phosphate Delivery System for Bone Morphogenetic Protein,” Clinical Orthopaedics and Related Research, 187: 277-280.
Kawamura, M., Iwata, H,. Sato, K. and Miura,T. (1987). “ Basic Science and Pathology: Chondroosteogenetic Response to Crude Bone Matrix Proteins Bound to Hydrozyapatite,” Clinical Orthopaedics and Related Research, 217: 281-292.
Deatherage, JR., and Miller, EJ. (1987). “ Packaging and Delivery of Bone Induction Factors in a Collagenous Implant,” Collagen Rel. Res., Vol. 7: 225-231.
Hotz, G., and Herr,.G. (1994). “ Bone Substitute With Osteoinductive Biomaterials - Current and Future Clinical Applications,” International Journal of Oral & Maxillofacial Surgery, Vol. 23: 413-417.
Hotz, G. and Herr, G. (1994),”Bone substitute with osteoinductive biomaterials current and further clinical application,”Int. J. Oral. Maxillofac.Surg. 23 : 413-417
Centrella, M., Horowitz MC., Wozney, J.M. and McCarthy T.L. (1994)”Transforming growth factor-beta gene family members and bone,” Endocr. Rev. 15: 27-39;
Robey, PG., Young, MF., Flander, KC. et al., (1994)”Osteoblasts synthesize and repond to transforming growth factor-beta in vitro,” J.Cell. Biol.105:201-205;
Bonewald, LF. and Dallas,SL. (1994)”Role of active and latent transforming growth factor beta in bone formation,”J.Cell.Biochem.55:35-357;
Miyazono,H., Ichijo,H., and Heldin,CH. (1993)”Transforming growth factor-beta : latent form, binding proteins and receptors,” Growth factor 8: 11-22;
Dallas, SL., Park-Snyder, S., Miyazono, K. et al., (1994)” Characterization and autoregulation of latent transforming growth factor-beta complexes in osteoblast-like cell lines. Production of a latent complex lacking the latent binding protein ,”J. Biol. Chem.269:6815-6821;
Pfeilschifter, J., D’Souza, SM., and Mundy, GR. (1987)” Effect of transforming growth factor-beta on osteoblastic osteosarcoma cells,”Endocrinology 121:212-218;
Rickard, DJ., Gowen, M. and MacDonald, BR. (1993)”Proliferation responces to estradiol, IL-1 alpha and TGF-beta by cells expressing alkaline phosphatase in human osteoblast-like cell culture,” Calcif. Tissue Int.52:227-233;
Canalis,E. (1985) “Effect of growth factors on bone cell replication and differentiation,” Clin Orthop.193:246-263;
Centrella, M., Casinghino, S., Ignotz, R. and MaCarthy, TL. (1992) “Multiple regulatory efects by transforming growth factor-beta on type- collagen levels in osteoblast-enriched cultures from fetal rat bone,” Endocrinology 131:2863-2872;
Wrana, JL., Maeno, M., Hawrylshyn, B., Yao, KL. and Sodek, J. (1998) ”Differentiation effects of TGF-beta on the synthesis of extracellular matrix proteins by normal fetal rat calvarial bone cell populations,”J.Cell.Biol.106:915-924;
Antosz, ME., Bellows, CG. and Aubin JE. (1989).”Effects of transforming growth factor-beta and epidermal growth factor on cell proliferation and the formation of bone nodules in isolated fetal rat calvaria cells,”J.Cell Physiol.140:386-395;
Noda, M. and Rodan, GA. (1986)”Type-beta transforming growth factor inhibits proliferation and expresion of alkaline phosphatase in murine osteoblast-like cells.”Biochem. Biophys. Res. Commun.140:56-65;
Wonzey, J.M.(1992).”The bone morphogenetic protein family and osteogenesis, ”Mol.Reprod. Dev.32 : 160-167
Celeste, AJ., Jannazzi, JA., Taylor, RC., Hewick, RM., Rosen, V., Wang, EA. and Wonzey, JM.(1990).”Identification of transforming growth factor-beta family members present in bone morphogenetic protein purified from bovin bone.”Proc. Natl. Acad. Sci. USA. 87 : 9843-9847
Amedee, J., Bareille, R., Rouais, F., Cunningham, N., Reddi, H. and Harmand, MF.(1994).”Osteogenin inhibits proliferation and stimulates differentiation of osteoprogenitors in human bone marrow,” Differentiation 58 : 157-164
Richard, DJ., Sullivan, TA., Shenker, BJ., Leboy, PS. and Hazhdan, I.(1994).”Induction of rapid osteoblast differentiation in rat bone marrow stromal cell culture by dexamathasone and BMP-2,” Dev.Biol. 161:218-228
Yasuharu, Y., Oida, S., Akimoto, Y. and Shioda, S.(1988).”Response of the Mouse Femoral Muscle to an Implant of a Composite of Bone Morphogenetic Protein and Plaster of Paris,” Clin Orthop.Vol.234 :240-249.
Peppas NA. (1987) “Time- and position-dependent drug delivery in controlled-release systems. “J. of Pharmal Sci. 76(3):267,.
Higuchi T.(1963).”Mechanism of Sustained-Action Medication .” J. of Pharma.Sci. 52:1145-1149
Fessi, H., Marty, JP., Puisieux, F. and Carstensen, JT. (1982). “Square Root of Time Dependence of Matrix Formulations with Low Drug Content,” Journal of Pharmaceutical Sciences, Vol. 71(7) : 749-752
Nava Saucedo, JE., and Barbotin, JN. (1993). “ Bioencapsulation Revisited,” Bioamt., Art. Cells & Immob. Biotech., Vol. 21(3): 383-389.
Jeng, MJ., Wang, YJ., and Tsai,.R.(1996). “ Reconstitution of Collagen in Alginate,” Institute of Biomedical Engineering, National Yang-Ming University, Taiwan, Master Thesis.
Miller, EJ. and Rhodes, RK.(1982) ”Preparation and Characterization of the Different Types of Collagen,” Methods in Enzymology Vol.82.. P.59-64
Kim, BS. and Mooney, DJ.(1998) “Development of biocompatible synthetic extracellular matrices for tissue engineering,” TIBTECH Vol.16 : 224-230
Marcel, EN. (1988). Collagen Vol.1 Biochemistry : 120-122
Lee, JM., Pereira, CA., Abdulla, D., Naimark, WA. and Crawford I. (1995). “A multi-sample denaturation temperature tester for collagenous biomaterials,” Med. Eng.Phys. Vol.17(2) : 115-121
Sarti,B. and Scandola, M.(1995).”Viscoelastic and thermal properties of collagen/ploy(vinyl alcohol) blends,” Biomaterials, 16 : 785-792
Lee,JM., Pereira,CA., Abdulla,D., Naimark,WA. And Crawford,.I.(1995).”A multi-sample denaturation temperature tester for collagenous biomaterials,”Med.Eng.Phys. 17 : 115-1221
Urist, MR., Iwata, H., Ceccotti, PL., Dorfman, RL., Boyd, SD., McDowell, RM. and Chien, C.. (1973). “ Bone Morphogenesis in Implants of Insoluble Bone Gelatin,” Proc. Nat. Acad. Sci. USA, Vol. 70(12): 3511-3515.
Lowry, OH., Rosebrough, NJ. Farr, AL and Randall, RJ. “J. Biol. Chem.” Vol. 193: P265, 1951.
Tanaka, H., Matsumura, M. and Veliky, IA.(1984)”Diffusion characteristics of substrates in Ca-alginate gel beads,” Biotech. Bioeng. 24:53-58
Pu, HT. and Yang, YK..(1988)”Diffusion of sucrose and yohimbine in calcium alginate gel beads with or without entrapped plant cells,” Biotech. Bioeng. 32:891-896
Mizutan,i H. and Urist, MR.(1982)”The nature of bone morphogenetic protein (BMP) fraction derived from bovine bone matrix gelatin,”Clinical Orthopaedics and Related Research, 171 : 213-223
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top