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Adipose-derived adult stem cells seeded in alginate-based nanocomposites combined with biodegradable precision scaffolds for cartilage tissue engineering
|關鍵字:||脂肪幹細胞;adipose-derived adult mesenchymal stem cells (ADAS);褐藻膠;氫氧基磷灰石;TGF-beta 3;PLGA精密支架;CBD-RGD;nanophase calcium-deficient hydroxy-apatite (nCDHA);alginate;TGF-beta 3;PLGA precision scaffolds;CBD-RGD||出版社:||化學工程學系所||引用:|| Hunter W. On the structure and diseases of articulating cartilage. Phil Trans 1743; 470: 514-521.  Buckwalter JA, Mankin HJ. Articular cartilage: degeneration and osteoarthritis, repair, regeneration, and transplantation. Instr Course Lect 1998; 47:487-504.: 487-504.  Messner K, Maletius W. The long-term prognosis for severe damage to weight-bearing cartilage in the knee: a 14-year clinical and radiographic follow-up in 28 young athletes. Acta Orthop Scand 1996; 67: 165-168.  Kim HK, Moran ME, Salter RB. The potential for regeneration of articular cartilage in defects created by chondral shaving and subchondral abrasion. An experimental investigation in rabbits. J Bone Joint Surg Am 1991; 73: 1301-1315.  Convery FR. The repair of large osteschondral defects. An experimentent study in horse. Clin Orthop 1972; 82: 253.  Meachim G. The effect of scartification on articular cartilage in rabbits. Journal of Bone and Joint Surgery; American volume 1963; 45: 150.  Peterson L. Autologous chondrocytes transplantation - long term results. Cartilage surgery and future perspectives: Springer 2003; 19-27.  Marlovits S, Zeller P, Singer P, Resinger C, Vecsei V. Cartilage repair: generations of autologous chondrocyte transplantation. Eur J Radiol 2006; 57: 24-31.  Sittinger M. Engineering cartilage structures. Cartilage surgery and future perspectives: Springer 2003; 67-74.  Malemud CJ, Papay RS. The in vitro cell culture age and cell density of articular chondrocytes alter sulfated-proteoglycan biosynthesis. J Cell Physiol 1984; 121: 558-568.  O''Driscoll SW. Preclinical cartilage repair: current status and future perspectives. Clin Orthop Relat Res 2001; S397-S401.  Chung C, Burdick JA. Engineering cartilage tissue. Adv Drug Deliv Rev 2008; 60: 243-262.  Paul de V, Faas MM, Strand B, Calafiore R. Alginate-based microcapsules for immunoisolation of pancreatic islets. Biomaterials 2006; 27: 5603-5617.  Skjak-Braek G, Grasdalen H, Larsen B. Monomer sequence and acetylation pattern in some bacterial alginates. Carbohydr Res 1986; 154:239-50.: 239-250.  Berth G. Methodical aspects of characterization of alginate and pectate by light scattering and viscometry coupled with GCP. Carbohydrate polymers 1992; 19: 1-9.  Rees DA.. Polysaccharide shapes and their interactions||摘要:||
本研究先製備一系列nCDHA/alginate奈米複合材料以三維培養方式，探討大鼠脂肪幹細胞經過ㄧ週與二週培養後，軟骨分化基因及軟骨基質分泌 (collagen，GAG) 進行評估，結果發現500 ppm nCDHA可增加第7天軟骨基因分化與第14天膠原蛋白增生。而在100 ppm nCDHA對細胞增生有顯著效果。更進一步，利用500 ppm nCDHA吸附生長因子 (TGF-beta 3) 結合褐藻膠製成之奈米複合材料以三維培養方式，探討大鼠脂肪幹細胞以含TGF-beta 3的軟骨分化劑培養ㄧ週與三週後，結果發現nCDHA吸附TGF-beta 3並以含TGF-beta 3的軟骨分化劑培養大鼠脂肪幹細胞，可增加第7天軟骨基因分化與第21天軟骨基質增生。進一步討論500 ppm nCDHA吸附TGF-beta 3結合alginate應用於PLGA精密支架 (precision scaffolds) 之三維培養，培養21天後，發現軟骨基質明顯增生。此外也評估nCDHA吸附TGF-beta 3/alginate結合CBD-RGD應用於PLGA精密支架，以含TGF-beta 3的軟骨分化劑培養大鼠脂肪幹細胞，可增加第21天軟骨基質增生。本研究建立了一個nCDHA/alginate/PLGA精密支架培養系統，可有效促進軟骨分化。
In this study, the effect of nCDHA on chondrogenic differentiation of rat adipose-derived adult mesenchymal stem cells (ADAS) in alginate gel was evaluated. nCDHA/alginate composites containing different amounts of nCDHA were prepared and used to encapsulate rat ADAS. The chondrogenic differentiation of ADAS was enhanced after 7 days and the total collagen content increased after 14 days at 500 ppm of nCDHA. Cell proliferation was promoted in alginate with 100 ppm of nCDHA. Furthermore, the presence of nCDHA, especially at 500 ppm, in the three dimensional alginate culture induced differentiation of rate ADAS toward chondrogenic phenotype. Furthermore, the combination of growth factor TGF-beta 3 adsorbed onto 500 ppm nCDHA and alginate was used to encapsulate rat ADAS as chondrogenic induction medium. The chondrogenic differentiation of ADAS was enhanced after 7 days and the total collagen content increased after 21 days. The addition of three-dimensional PLGA precision scaffolds and CBD-RGD to chondrogenic induction medium had the same effect on the total collagen content, which increased after 21 days. In this research, we had developed an effective three-dimension culture system for chondrogenesis.
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