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研究生:胡淑文
研究生(外文):Shu-Wen Whu
論文名稱:幾丁聚醣/動物明膠複合物作為軟骨再生支架之研究
論文名稱(外文):Assessment of Chitosan/ Gelatin Complexes as Tissue Engineering Scaffolds for Cartilage Regeneration
指導教授:徐善慧徐善慧引用關係
學位類別:博士
校院名稱:國立中興大學
系所名稱:化學工程學系所
學門:工程學門
學類:化學工程學類
論文種類:學術論文
畢業學年度:96
語文別:英文
論文頁數:116
中文關鍵詞:幾丁聚醣動物明膠軟骨組織工程動態機械分析TGF-β3人類骨髓間質幹細胞動態培養
外文關鍵詞:Chitosangelatincartilagetissue engineeringdynamic mechanical analysisTGF-β3human bone marrow mesenchymal stem cellsdynamic culture
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軟骨組織工程中,基於結合軟骨細胞和生醫材料的細胞支架一直不斷在開發。其中最重要的成功關鍵就是找到最佳的材料作成支架。本篇將利用幾丁聚醣-動物明膠聚電性複合材料作為軟骨組織工程支架進行體外和體內相關測試。自戊二醛、雙環氧化物和水溶性氰胺物等三種交聯劑中找到水溶性氰胺物是動物明膠的最佳交聯劑。當幾丁聚醣和動物明膠比例為1:1時複合支架有適當的降解速率與機械穩定性。以水溶性氰胺物交聯後的幾丁聚醣-動物明膠聚電性複合材料有很好的軟骨細胞增殖和細胞間質分泌表現。進行兔子體內實驗後,幾丁聚醣-動物明膠聚電性複合材料仍然確定為適合做為軟骨組織再生支架。
為了獲得更佳的體外培養條件,利用旋轉型生物反應器提供良好的質傳與生理上的翦應力,使得再生軟骨有好的機械強度和壓縮模數。但是動態培養並不是產生類真實組織結構之新生軟骨的必要條件。另外,發現動物明膠能促進人類軟骨細胞增殖,並且幾丁聚醣能維持人類軟骨細胞型態。
為了增加軟骨細胞來源,將人類骨髓間葉幹細胞(hBMSC)植入包括第二型膠原蛋白改質的PLGA/PLLA支架(BCII)與幾丁聚醣/動物明膠混合支架(CG) 的兩種三維支架。結果發現CG支架內的細胞數量較BCII支架為高。支架種類並不影響TGF-β3促進骨髓間葉幹細胞之分化;動態培養並不幫助骨髓間葉幹細胞之分化。
最後,目前沒有任何方法能夠對組織工程軟骨進行非破壞性的品質控管。本研究研發利用動態機械分析儀和流變分析來確認不同部位的軟骨組織 (如關節、肋骨和耳朵)。不同部位的軟骨有不同的storage modulus。當軟骨組織能負載應力高則有較高的storage modulus表現。而關節和肋骨等屬於透明軟骨等組織有相近的loss tan。另ㄧ方面,來自耳朵的彈性軟骨表現出有別於關節和肋骨的loss tan。儘管耳朵軟骨組織有高於關節軟骨的基質含量和細胞數量,機械性質卻遠低於關節軟骨。而組織工程細胞支架經由1和28天的培養後,細胞數量、細胞間質含量和storage modulus會隨時間增加不過仍遠低於真實軟骨組織但是細胞支架的loss tan卻接近真實組織。所以loss tan可以作為組織工程軟骨的品管參數。
Constructs based on chondrocytes and biomaterial scaffolds were developed for cartilage tissue engineering. One of the keys for success is to select suitable materials for fabrication of the scaffolds. In this study, chitosan-gelatin polyelectrolyte complexes were evaluated as tissue engineering scaffolds for cartilage regeneration in vitro and in vivo. The crosslinker for gelatin was selected among glutaradehyde, bisepoxy and water-soluble carbodiimide (WSC), based upon the growth of chondrocytes on the crosslinked gelatin. WSC was found to be the most ideal crosslinker for the system. The complex scaffolds with chitosan/gelatin ratio equal to one possessed the proper degradation profile and mechanical stability. Chondrocytes proliferated well and secreted extracellular matrix in chitosan-gelatin complex scaffolds crosslinked by WSC. Rabbit implantation confirmed the suitability of using chitosan-gelatin complex scaffolds for cartilage tissue engineering.
For better culture condition, it was hypothesized that good mass transfer and the physiological shear provided by the rotating-type bioreactor were important for the neocartilage formed in the scaffolds to exhibit satisfactory mechanical strength and compression modulus; However, the dynamic culture condition was not prerequisite for the constructs to develop a histological resemblance to the real tissues. Then gelatin was observed to promote the human chondocytes proliferation; while chitosan was observed to maintain the human chondocytes morphology.
For more cell source, human bone marrow mesenchymal stem cells (hBMSC) were seeded into two scaffolds, including blended polymers of PLGA50/50 and PLLA modified by type II collagen (BCII) and chitosan-gelatin complexes (CG). Cell numbers in CG scaffolds were higher than those in BCII scaffolds. The materials of the scaffolds had no effect on TGF-β3 induced hBMSC transformation into differentiated cells. The dynamic culture system promoted cell proliferation, but not cell differentiation.
Finally, there is no current method for non-destructive quality control of tissue-engineered cartilage. This study explored a way to utilize a dynamic mechanical analyzer and rheological analysis to assess the cartilage tissues from different anatomic locations (e.g. arthrosis, costa and ear). Cartilage from different locations showed different storage modulus. Higher storage modulus was observed in positions that offered a greater loading force. Hyaline cartilage, either from arthrosis or costa, had similar values in loss tan. On the other hand, elastic cartilage (from ear) showed a distinct value of loss tan from that of arthrosis or costa. In spite of the much higher matrix content and cell number for ear cartilage vs. arthrosis cartilage, the mechanical properties of ear cartilage were much lower than those of arthrosis cartilage. Tissue-engineered constructs were cultured for 1 and 28 days, where the cell number, matrix content and storage modulus all increased with the culture time, but were still much lower than the values in the real cartilage. The values of loss tan of all constructs, however, approached those of real cartilage. It thus appeared that values of loss tan may serve as one of the major performance indice for tissue-engineered cartilage.
I. Chinese abstract (中文摘要) 1
II. Abstract 3
III. Acknowledgment (謝誌) 6
IV. Contents 7
V. Figures 11
VI. Tables 13
VII. Abbreviation 14
Chapter 1. Neocartilage Formation in Chitosan-Gelatin Scaffolds: In Vitro and In Vivo Studies 16
1.1. Introduction 16
1.2. Materials and methods 18
1.2.1. Sample preparation 18
1.2.2. Sample characterization 20
1.2.3. Cell growth in vitro 22
1.2.4. Animal implantation 24
1.2.5. Statistical analysis 25
1.3. Results 25
1.3.1. Determination of crosslinking condition for gelatin 26
1.3.2. Characterization of scaffolds 27
1.3.3. Three-dimensional cell growth 29
1.3.4. In vivo implantation 30
1.4. Discussion 31
1.5. Conclusion 35
1.6. References 36
Chapter 2. Neocartilage formation in two scaffolds in vitro 43
2.1. Introduction 43
2.2. Materials and methods 44
2.2.1. Cells preparation 44
2.2.2. Cell growth in vitro 44
2.2.3. Statistical analysis 45
2.3. Results 46
2.4. Discussion 47
2.5. Conclusion 48
2.6. References 48
Chapter 3. Evaluation of bone marrow mesenchymal stem cells and composite scaffolds with dynamic culture system for neo-cartilage regeneration in vitro 50
3.1. Introduction 50
3.2. Materials and methods 51
3.2.1. Isolation of bone marrow mesenchymal stem cell 51
3.2.2. Induction of bone marrow mesenchymal stem cell Cell growth in vitro 51
3.2.3. Statistical analysis 52
3.3. Results 53
3.3.1. The transformation of BMSCs in BCII scaffolds 53
3.3.2. The transformation of BMSCs in CG scaffolds 53
3.3.3. The transformation of BMSCs in CG scaffolds under dynamic culture 54
3.4. Discussion 55
3.5. Conclusion 56
3.6. References 56
Chapter 4. Rheological Properties of Cartilage 58
4.1. Introduction 58
4.2. Materials and methods 60
4.2.2. Dynamic mechanical analysis 60
4.2.3. Biochemical content analysis 61
4.2.4. Preparation of tissue engineering constructs 61
4.2.5. Statistical analysis 63
4.3. Results 63
4.3.1. Compressive mechanical properties 63
4.3.2. Tensile mechanical properties 64
4.3.3. Biochemical contents 64
4.3.4. Mechanical properties of tissue- engineered constructs 65
4.4. Discussion 65
4.5. Conclusion 67
4.6. References 68
Chapter 5. Conclusion 72
Appendix A. Culture of human cartilage chondrocytes on different materials ……………………………………………………………100
A.1. Introduction 100
A.2. Materials and methods 100
A.3. Results 101
A.4. Discussion 102
A.5. Conclusion 102
A.6. References 103
Appendix B. Cell density of fresh cartilage tissue in rabbit’s costa and ear…… 110
Appendix C. Journal publication 111
Appendix D. Conference paper publication 113
Appendix E. Prize 116
Chapter 1
1.Hunter, W. Of the structure and diseases of articulating cartilages, Philosophical Transactions 470:514-21, 1743.
2.Buckwalter, J.A., and Mankin, H.J. Articular cartilage repair and transplantation. Arthrit Rheum 41 (8):1331-42, 1998.
3.Buckwalter, J.A., and Mankin, H.J. Articular cartilage. Part I: tissue design ad chondrocyte-matrix interactions, J Bone Joint Surg 79a:600, 1997.
4.Freeman, M.A.R. Adult articular cartilage, Oxford: Alden Press, 1973.
5.Mitchell, N., and Shepard, N. The resurfacing of adult articular cartilage by multiple perforation through the subchondral bone, J Bone Jt Surg 58a(2): 230-3, 1976.
6.Francis Suh, J.K., and Matthew, H.W.T. Application of chitosan-based polysaccharide biomaterials in cartilage tissue engineering: a review. Biomaterials 21:2589-2598, 2000.
7.Tsai, C.L., Hsu, S., and Tang, C.M. Evaluation of the Cellular Adhesion and Growth on Biodegradable Polymers Using Immortalized Rat Chondrocytes. Artif Organs 26(7):647-658, 2002.
8.von Schroeder, H.P., Kwan, M., Amiel, D., and Coutts, R.D. The use of polylactic acid matrix and periosteal grafts for the reconstruction of rabbit knee articular defects. J Biomed Mater Res 25:329-39, 1991.
9.Freed, L.E., Marquis, J.C., Nohria, A., Emmanual, J., Mikos, A.G., and Langer, R. Neocartilage formation in vitro and in vivo using cells cultured on synthetic biodegradable polymers. J Biomed Mater Res 27:11-23, 1993.
10.Ishaug-Riley, S.L., Okun, L.E., Prado, G., Applegate, M.A., and Ratcliffe, A. Human articular chondrocyte adhesion and proliferation on synthetic biodegradable polymer films. Biomaterials 20:2245-46, 1999.
11.Mikos, A.G., McIntire, L.V., Anderson, J.M., and Babensee, J.E. Host response to tissue engineered devices. Adv Drug Deliv Rev 33:111-139, 1998.
12.Madihally, S.V., and Matthew, H.W.T. Porous chitosan scaffolds for tissue engineering. Biomaterials 20, 1133, 1999.
13.Shapiro, L., and Cohen, S. Novel alginate sponges for cell culture and transplantation. Biomaterials 18, 583, 1997.
14.Toolan, B.C., Frenkel, S.R., Pachence, J.M., Yalowitz, L., and Alexander, H. Effects of growth-factor-enhanced culture on a chondrocyte-collagen implant for cartilage repair. J Biomed Mater Res 31, 273, 1996.
15.Grande, D.A., Halberstadt, C., Naughton, G., Schwartz, R., and Manji, R. Evaluation of matrix scaffolds for tissue engineering articular cartilage. J Biomed Mater Res 34, 211, 1997.
16.Pieper, J.S., van der Kraan, P.M., Hafmans, T., Kamp, J., Buma, P., van Susante, J.L.C., van den Berg, W.B., Veerkamp, J.H., and van Kuppevelt, T.H. Crosslinked type II collagen matrices: preparation, characterization, and potential for cartilage engineering. Biomaterials 23, 3183, 2002.
17.Amiel, G.E., Komura, M., Shapira, O., Yoo, J.J., Yazdani, S., Berry, J., Kaushal, S., Bischoff, J., Atala, A., and Soker, S. Engineering of blood vessels from acellular collagen matrices coated with human endothelial cells. Tissue Eng 12(8):2355-65, 2006.
18.Huynh, T., Abraham, G., Murray, J., Brockbank, K., Hagen, P.O., and Sullivan, S. Remodeling of an acellular collagen graft into a physiologically responsive neovessel. Nat Biotechnol 17(11):1083-6, 1999.
19.Aigner, J., Tegeler, J., Hutzler, P., Campoccia, D., Pavesio, A., Hammer, C., Kastenbauer, E., and Naumann, A. Cartilage tissue engineering with novel nonwoven structured biomaterial based on hyaluronic acid benzyl ester. J Biomed Mater Res 42, 172, 1998.
20.Tomihata, K., Burczak, K., Shiraki, K., and Ikada, Y. Cross-linking and biodegradation of native and denatured collagen. ACS Symposium Series No.540.
21.Hodde, J. Naturally occurring scaffolds for soft tissue repair and regeneration. Tissue Eng 8(2): 295-308, 2002.
22.Lu, J.X., Prudhommeaux, F., Meunier, A., Sedel, L., and Guillemin, G. Effect of chitosan on rat knee cartilages. Biomaterials 20:1275-1285, 1999.
23.Sechriest, V.F., Niyibizi, Y.J., Westerhausen-Lason, A., Matthew, H.W.T., Evans, C.H., Fu, F.H., and Francis Suh, J.K. GAG-augmented polysachharide hydrogel: a novel biocompatible and biodegrable materials to support chondrogenesis. J Biomed Mater Res 49:534-41, 2000.
24.Takahashi, O., Takayama, K., Machida, Y., and Nagai, T. Characteristics of polyion complexes of chitosan with sodium alginate and sodium polyacrylate. Int J Pharm 61:35-41, 1990.
25.Gaserod, O., Smidsrod, O., and Skjak-Braek, G. Microcapules of alginate-chitosan-chitosan-I. A quantitative study of the interaction between aginate and chitosan. Biomaterials 19(20): 1815-25, 1998.
26.Denuiziere, A., Ferrier, D., Damour, O., and Domard, A. Chitosan-chondroitin sulfate and chitosan-hyaluronate polyelectrolyte complexes:biological properties. Biomaterials 19:1275-1285, 1998.
27.Hsu, S., and Jamieson, A.M. Viscoelastic behavior at the thermal solgel transition of gelatin. Polymer 34, 2602, 1993.
28.Winter, H. Can the gel point of a cross-linking polymer be detected by G’-G’’ crossover? Polymer Engineering and Science 27, 1698, 1987.
29.Horton, W.E.J., Cleveland, J., Rapp, U., Nemuth, G., Bolander, M., Doege, K., Yamada, Y., and Hassell, J.R. An established rat cell line expressing chondrocyte properties. Exp Cell Res 178:457-68, 1988.
30.Freshney, R.I. Culture of animal cells: a manual of basic technique. 3rd edition. New York: Wiley-Liss, 331-2, 1994.
31.Kim, Y.J., Sah, R.Y., Doong, J.Y., and Grodzinsky, A.J. Fluorometric assay of DNA in cartilage explants using Hoechst 33258. Anal Biochem 174:168-176, 1988.
32.Enobakhare, B.O., Bader, D.L., and Lee, D.A. Quantification of sulfated glycosaminoglycans in chondrocyte/alginate culture by use of 1,9- dimethylmethylene blue. Anal Biochem 243:189-191, 1996.
33.Hsu, S., Whu, S.W., Hsieh, S., Tsai, C., Chen, D.C., and Tan, T. Evaluation of chitosan-alginate- hyaluronate complexes modified by an RGD-containing protein as tissue engineering scaffolds for cartilage regeneration. Artif Organs 28:693-703, 2004.
34.Mao, J.S., Zhao, L.G., Yin, Y.J., and Yao, K.D. Structure and properties of bilayer chitosan-gelatin scaffolds. Biomaterials 24:1067-1074, 2003.
35.Lee, J.H., and Lee, H.B. A wettability gradient as a tool to study protein adsorption and cell adhesion on polymer surface. J Biomater Sci Polym Ed 5:467-81, 1993.
36.Matsumoto, T., Kawai, M., and Masuda, T. Rheological properties and fractal structure of concentrated polyion complexes of chitosan and alginate. Biorheology 30:435-441, 1993.
37.Freed, L.E. Tissue engineering of cartilage. The Biomedical Engineering Handbook. Bronzino, J. D. (ed.), CRC Press, Boca Raton, chap. 124, 2000.
Chapter 2
1.Freed LE, Vunjak-Novakovic G, Culture of organized cell communities, Advanced Drug Delivery Review, 1998; 33;15-30
2.Tsai CL, Hsu S, and Tang CM, Evaluation of the Cellular Adhesion and Growth on Biodegradable Polymers Using Immortalized Rat Chondrocytes, Artif Organs, 2002; 26(7):647-658.
3.Freshney RI, Culture of animal cells: a manual of basic technique, 3rd edition. New York: Wiley-Liss, 1994:331-2
4.Kim YJ, Sah RY, Doong JY, Grodzinsky AJ. Fluorometric assay of DNA in cartilage explants using Hoechst 33258. Anal Biochem 1988;174:168-176.
5.Enobakhare BO, Bader DL, Lee DA. Quantification of sulfated glycosaminoglycans in chondrocyte/alginate culture, by use of 1,9-dimethylmethylene blue. Anal Biochem 1996;243:189-191.
6.Bergman M, Loxley R. Two improved and simplified methods for the spectrophotometric determination of hydroxyproline. Anal Biochem 1961;35(12):1961-1965.
Chapter 3
1.Majumdar MK, Banks V, Pelus DP, and Morris EA, J. Cell. Physiol. 185:98–106, 2000
2.Tsai CL, Hsu, S, and Tang C-M, Artif Organs. 26(7): 647-658, 2002
3.Whu SW, Hsu S, Tsai CL, and Hsieh KH, J. Engineering National Chung Hsing University, Vol.12, No.2, pp.103-108, 2001
4.Freed LE, and Vunjak-Novakovic G, Tissue Engineering of Cartilage, In The Biomedical Engineering Handbook 2nd ed Vol. II, Bronzino JD (ed.), CRC Press, Boca Raton , FL, 2000, Chap.124
chapter 4
1.S. Treppo, H. Koepp, E.C. Quan, A.A. Cole, K.E. Kuettner and A. J. Grodzinsky, Comparison of biomechanical and biochemical properties of cartilage from human knee and ankle pairs, J. Orthop. Res. 18 (2000), 739-48.
2.J.S. Jurvelin, M.D. Buschmann and E.B. Hunziker, Mechanical anisotropy of the human knee articular cartilage in compression, Proc. Inst. Mech. Eng. [H] 217 (2003), 215-9.
3.D.D. Anderson, T.D. Brown, K.H. Yang and E.L. Radin, A dynamic finite element analysis of impulsive loading of the extension-splinted rabbit knee, J. Biomech. Eng. 112 (1990), 119-28.
4.L.J. Bonassar, K.A. Jeffries, E.H. Frank, V.L. Moore, M.W. Lark, E.K. Bayne, J. McDonnell, J. Olszewski, W. Hagmann and K. Chapman, In vivo effects of stromelysin on the composition and physical properties of rabbit articular cartilage in the presence and absence of a synthetic inhibitor, Arthritis Rheum. 38 (1995), 1678-86.
5.E.A. Kennedy, D.S. Tordonado and S.M. Duma, Effects of freezing on the mechanical properties of articular cartilage, Biomed. Sci. Instrum. 43 (2007), 342-7.
6.J.B. Choi, I, Youn, L. Cao, H.A. Leddy, C.L. Gilchrist, L.A. Setton and F. Guilak, Zonal changes in the three-dimensional morphology of the chondron under compression: the relationship among cellular, pericellular, and extracellular deformation in articular cartilage, J. Biomech. 40 (2007), 2596-603.
7.L.J. Bonassar, E.H. Frank, J.C. Murray, C.G. Paguio, V.L. Moore, M.W. Lark, J.D. Sandy, J.J. Wu, D.R. Eyre and A.J. Grodzinsky, Changes in cartilage composition and physical properties due to stromelysin degradation, Arthritis Rheum. 38 (1995),173-83.
8.G. Spahn, E. Kahl, H.M. Klinger, T. Mückley, M. Günther and G.O. Hofmann, Mechanical behavior of intact and low-grade degenerated cartilage, Biomed. Tech. 52 (2007), 216-22.
9.M. Baldassarri, J.S. Goodwin, M.L. Farley, B.E. Bierbaum, S.R. Goldring, M.B. Goldring, D. Burstein and M.L. Gray, Relationship between cartilage stiffness and dGEMRIC index: correlation and prediction, J. Orthop. Res. 25 (2007), 904-12.
10.S. Hsu, S.W. Whu, S. Hsieh, C. Tsai, D.C. Chen and T. Tan. Evaluation of Chitosan-Alginate-Hyaluronate Complexes Modified by an RGD- containing Protein as Tissue Engineering Scaffolds for Cartilage Regeneration, Artificial Organs, 28 (2004), 693-703.
11.S. Hsu, C. Kuo, H. Yen, S.W. Whu and C. Tsai. The effect of two different bioreactors on the neocartilage formation in type II collagen modified polyester scaffolds seeded with chondrocytes, Artificial Organs, 29 (2005), 467-74.
12.S. Hsu, S.H. Chang, H. Yen, S.W. Whu, C. Tsai and D.C. Chen. Evaluation of biodegradable polyesters modified by type II collagen and Arg-Gly-Asp as tissue engineering scaffolding materials for cartilage regeneration, Artificial Organs, 30 (2006), 42-55.
13.Y.J. Kim, R.Y. Sah, J.Y. Doong and A.J. Grodzinsky. Fluorometric assay of DNA in cartilage explants using Hoechst 33258. Anal. Biochem. 174 (1988), 168-176.
14.B.O. Enobakhare, D.L. Bader and D.A. Lee. Quantification of sulfated glycosaminoglycans in chondrocyte/alginate culture, by use of 1,9-dimethylmethylene blue, Anal. Biochem. 243 (1996), 189-191.
15.M. Bergman and R. Loxley. Two improved and simplified methods for the spectrophotometric determination of hydroxyproline, Anal. Biochem. 35 (1961), 1961-1965.
16.K. Tomihata, K. Burczak, K. Shiraki and Y. Ikada. Cross-linking and biodegradation of native and denatured collagen, ACS Symposium Series No.540.
Appendix A
1.Francis Suh, J. K. and Matthew, H. W. T., Biomaterials, 21, pp.2589-2598, 2000.
2.Hye-Won, Yasuhiko Tabata and Yoshito Ikada, Biomaterials, 20, pp. 1339-1344, 1999.
3.Ross Tubo and Francois Binettem, Methods in Molecular Medicine, Vol. 18: Tissue Engineering Methods and Protocols pp.205-215, 1999.
4.Shan-hui Hsu, Chi-Ching Kuo, Shu Wen Whu, Chen-Huan Lin,and Ching-Lin Tsai. The effect of ultrasound stimulat1h3ion versus bioreactors on neocartilage formation in tissue engineering scaffolds seeded with human chondrocytes in vitro. Biomol Eng, Oct;23(5):259-64, 2006
5.Muller-Rath R, Gavénis K, Andereya S, Mumme T, Schmidt-Rohlfing B, Schneider U. A novel rat tail collagen type-I gel for the cultivation of human articular chondrocytes in low cell density. Int J Artif Organs Dec;30(12):1057-67, 2007.
6.Lin YJ, Yen CN, Hu YC, Wu YC, Liao CJ, Chu IM. Chondrocytes culture in three-dimensional porous alginate scaffolds enhanced cell proliferation, matrix synthesis and gene expression. J Biomed Mater Res A. Feb 6, 2008.
7.Yen CN, Lin YR, Chang MD, Tien CW, Wu YC, Liao CJ, Hu YC. Use of porous alginate sponges for substantial chondrocyte expansion and matrix production: effects of seeding density. Biotechnol Prog. Mar-Apr;24(2):452-7, 2008.
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