本研究利用連續性微溝槽的聚二甲基矽氧烷（polydimethalsiloxane，PDMS）來探討人類牙齦纖維母細胞（human gingival fibroblasts，hGF）的行為表現。微溝槽的寬度由1μm至20μm，而深度維持於1μm。人類牙齦纖維母細胞的排列性（alignment）及延展性（elongation）指出人類牙齦纖維母細胞在微溝槽上具有接觸引導（contact guidance）的現象。細胞的排列性及延展性會隨著微溝槽寬度的減少而有所增加；然而，細胞延展面積則是隨著微溝槽寬度的增加而增加。微溝槽寬度較大的區域有較高的細胞貼附及增生表現，而平滑區域則有最高的細胞密度。以上結果與類骨母細胞的貼附、增生趨勢有所差異，而這與不同細胞的延伸能力(stretching ability)有關。電子顯微鏡顯示當微溝槽寬度小於10μm，hGF無法觸及微溝槽底部，而類骨母細胞則能輕易觸及微溝槽底部。也就是說細胞勁度（cell stiffness）可能限制了細胞的延伸能力，具有較高細胞勁度的細胞可能無法跨越過微溝槽邊緣，但可以沿著微溝槽方向生長，這可以由微溝槽表面有較高程度的細胞排列性得知。表面型態會促進鹼性磷酸酶表現，而狹窄的表面溝槽會有最強烈的鹼性磷酸酶表現，此結果與類骨母細胞（ROS cells）的結果相同。對照細胞貼附及鹼性磷酸酶表現結果，微溝槽結構對於骨母細胞的骨誘導性是有所助益；而微溝槽結構對人類牙齦纖維母細胞則有助於細胞骨架（cytoskeleton）的再排列（re-arrangement）。

In this study, human gingival fibroblasts (hGF) cells were cultured onto PDMS surface which has characteristics of continuous groove width varied from 1μm to 20μm. The depth of grooves was kept as 1μm. Phenomenon of “contact guidance” was found in the culture system on microgrooved substrates, which was indicated by the alignment and elongation of hGF. By decreasing the groove width, degree of alignment and elongation would be increased on it. On the other hand, the cells spreading area would be larger with the increase in the groove width. The cell attachment and proliferation were both higher on the surface with larger groove width, while the highest cell densities were found on the smooth surface. The above results did not agree with the attachment and proliferation tendency of osteoblast-like cells; that was, the decrease in groove width would increase cell density. The reason of these findings may come from the divergence of stretching ability from different cells. From SEM images, it would be difficult for hGF to reach the bottom when the groove width was smaller than 10μm, but the osteoblast-like cells could easily reach the bottom of groove with smallest width. Namely, the cells stiffness might restrict their stretching ability. With high stiffness, cells would not cross over the ridge texture but adjusted to the grooves direction. It was shown by the high degree of cell alignment on grooves surface. The ALP expression would be accelerated by surface topography, where the strongest expression would be found on narrow pattern. This finding was in agreement with osteoblast-like ROS cells. Correlating with the contrast findings in attachment, from the ALP expression suggested that grooved pattern might be effectively in contributing osteoconduction of osteoblast cells; while in hGF cells, grooved pattern would be effective in re-arrangement of cytoskeleton.

Albelda SM, Buck CA, Integrins and other cell adhesion molecules, FASEB J., 1990; 4: 2868-2880
American Association of Oral and Maxillofacial Surgeons, 2008, Dental Implants, retrieved July 9, 2009, from http://www.aaoms.org/dental_implants.php
Arias C, (2007), What causes tooth loss?, retrieved July 7, 2009, from http://www.orlandodenturesandimplants.com/tooth-loss/causes-diseases
BH Jo, van Lerberghe LM, Motsegood KM, Beebe DJ, Three-dimensional micro-channel fabrication in polydimethylsiloxane (PDMS) elastomer, J Microelectromech Syst, 2000; 9: 76-81
Bennet JH, Carter DH, Alavi AL, Beresford JN, Walsh S, Patterns of integrin expression in a human mandibular explant model of osteoblast differentiation, Arch Oral Biol, 2001; 46: 229-238
Bissell MJ, Weaver VM, Lelievre SA, Wang F, Petersen OW, and Schmeichel KL, Cancer Res, 1999; 59: 1757–1763
Bissell MJ, Rizki A and Mian IS, Curr. Opin. Cell Biol, 2003; 15: 753–762
Bosman Fred T, Stamenkovic I, Functional structure and composition of the extracellular matrix, Journal of Pathology, 2003; 200: 423-428
Brunette DM, Kenner GS, Gould TRL, Grooved titanium surfaces orient growth and migration of cells from human gingival explants, J. Dent. Res., 1983; 62: 1045-1048
Brunette DM, Oakley C, Jaeger, Nicolas AF, Sensitivity of Fibroblasts and their cytoskeletons to Substratum Topographies: Topographic Guidance and Topographic Compensation by Micromachined Grooves of Different Dimensions, Experimental Cell Research, 1997; 234: 413-424
Chaudhury MK, J Adhes Sci Technol, 1993; 7: 669; Chaudhury MK, Biosens Bioelectron, 1995; 10: 785
Chen CS, Mrksich M, Huang S, Whitesides GM and Ingber DE, Science,1997; 276: 1425–1428
Chen J, Wolke G.C, de Groot K, Biomaterials, 1994; 15: 396
Chou L, Brunette DM, Firth James D, Uitto Veli-Jukka, Substratum surface topography alters cell shape and regulates fibronectin mRNA level, mRNA stability, secretion and assembly in human fibroblasts, Journal of Cell Science, 1995; 108: 1563-1573
Clark P, P Connolly, ASG Curtis, JAT Dow, CDW Wilkinson, Topographical control of cell behavior: II. Multiple grooved substrata, Development, 1990; 108: 635 – 644
Cowles EA, Brailey LL, Gronowicz GA, Integrin-mediated signaling regulates AP-1 transcription factors and proliferation in osteoblasts, J Biomed Mater Res, 2000; 52: 725-737
Curtis ASG, Varde M, Control of cell behavior: topological factors, JNCI, 1964; 33: 15-26
Dalby MJ, Riehle MO, Yarwood Stephen J, Wilkinson CDW, Curtis, ASG, Nucleus alignment and cell signaling in fibroblasts: response to a micro-grooved topography, Experimental Cell Research, 2003; 284:274-282
Defilippi P, Olivo C, Venturino M, Dolce L, Silengo L, Tarone G, Actin cytoskeleton organization in response to integrin-mediated adhesion, Micr Res Tech, 1999; 47: 67-78
den Braber ET, de Ruijter JE, Jansen JA, The effect of a subcutaneous silicone rubber implant with shallow surface microgrooves on the surrounding tissue in rabbits, J Biomed Mater Res, 1997; 37: 539-547
Deutsch J, Motlagh D, Russell B, Desai TA, Fabrication of microtextured membranes for cardiac myocyte attachment and orientation, J Biomed Mater Res, 2000; 53: 267- 275
dos Remedios CG, Chhabra D, Kekic M, Dedove IV, Tsubakihara M, Berry D A, Nosworthy N J, Actin binding proteins: regulation of cytoskeletal microfilaments, Physiol, 2002; 83: 433-473
Dunn GA, Heath JP, A new hypothesis of contact guidance in tissue cells, Exp. Cell Res., 1976; 101: 1-14
Dunn GA, Brown AF, Alignment of fibroblasts on grooved surfaces described by a simple geometric transformation, J. Cell Sci., 1986; 83: 313 – 340

Feng HL, Zhao BH, Bai W, Lee I-S, Cui FZ, Improvement of fibroblast adherence to titanium surface by calcium phosphate coating formed with IBAD, Surface &Coatings Technology, 2005; 193: 366-371
Feng Y, Walsh CA, The many faces of filamin: a versatile molecular scaffold for cell motility and signaling, Nat. Cell Biol, 2004; 6: 1034–1038
Flemming RG, Murphy CJ, Abrams GA, Goodman SL, Nealey PF, Effects of micro- and nano- structured surfaces on cell behavior, Biomaterials, 1999; 20: 573-588
Fransiska Swary, The Effects of Surface Micro-Patterns on Osteoblastic cells' Behaviors, (Master Thesis, University of Science and Technology, 2007)
Fritz JL, Owen MJ, J Adhesion, 1995; 54: 33
Fu Ricky KY, Chu Paul KH, Plasma modification of materials, Encyclopedia of Biomaterials and Biomedical Engineering, 2005; DOI:10.1081/E-EBBE-120007373
Guo, WJ, FG. Giancotti, Integrin signaling during tumor progression. Nature Reviews Molecular Cell Biology, 2004; 5(10):816-826
Gronowicz G, McCarthy MB, Response of human osteoblasts to implant materials: integrin-mediated adhesion, J Orthop Res, 1996; 14: 878-887
Grossner-Schreiber B, Griepentrog M, Haustein I, Muller W-D, Lange K-P, Briedigkeit H, Gobel UB, Plaque formation on surface modified dental implants, Clinical Oral Implant Research, 2001; 12: 543-551
Gougaloff R, 2005, Types of dental implants, retrieved july8, 2009, from http://www.laimplants.net/FredStalley_DI_Types.htm#top
Harrison RG, On the stereotropism of embryonic cells, Science, 1911; 34: 279
Hoebergen A., Uyama Y, Okada T, Ikada Y, Appl Polym Sci, 1993; 48: 1825
Huck TSW, Bowden N, Onck P, Pardoen T, Huychinson JW, Whitesides GM, Langmuir, 2000; 16: 3497
Hynes RO, Integrins: versatility, modulation, and signaling in cell adhesion, Cell, 1992; 69: 11-25
Iwata H, Kishida A, Suzuki M, and Ikada Y, J polym Sci, Polym Chem Ed, 1988; 26: 3309
Ishii M, Komatsubara M, in proceedings,1998 IEEE Conf on Electrical Insulation and Dielectric Phenomena, Atlanta, 1998;134
Jansen, JA, Siebers MC, ter Brugge PJ, Walboomers XF, Integrins as linker proteins between osteoblasts and bone replacing materials. A critical review, Biomaterials, 2005; 26: 137-146
Jessamine Ng Lee, Cheolmin Park, George M, Whitesides, Solvent Compatibility of Poly(dimethylsiloxane)-Based Microfluidic Devices, Anal Chem, 2003; 75: 6544-6554

Kaplowitz GJ, n.d., I2 Screw Type Groovy Dental Implants, retrieved July 9, 2009, from http://www.osseonews.com/drosseo/content/78646-i2-screw-type-groovy-dental-implants
Kim B, Peterson ETK, Papautsky I, Long-Term Stability of Plasma Oxidized PDMS Surfaces, Proceedings of the 26th Annual International Conference of the IEEE EMBS, San Fransisco, CA, USA, September 1-5, 2004
Kim E, Xia Y and Whitesides G M, Nature, 1995; 376: 581
Kim E, Xia Y, Zhao X-M, Whitesides G M, Adv Mater, 1997; 9: 651
Kim H, Sengupta A, Glogauer M, Mc Culloch CA, Filamin A regulates cell spreading and survival via β1 integrin, Experimental Cell Research, 2008; 314: 834-846
Lee JN, Park CM, Whitesides GM, Solvent compatibility of poly-(dimethylsiloxane)-based microfluidic devices, Anal. Chem., 2003; 75: 6544-6554
Lesol H, Osman M, Ruch JV, Immunofluorescent localizations of collagens, fibronectin, and laminin during terminal differentiation of odontoblasts, Developmental Biology, 1981; 82: 371-81
Lin W-C, Yang M-C, Protein adsorption and platelet adhesion of polysulfone membrane immobilized with chitosan and heparin conjugate, Polym Adv Technol, 2003; 14: 103-113
Liston EM, Martinu L, Wertheimer MR, in plasma Surface Modification of Polymers: Relevance to Adhesion, (M Strobel, C Lyons, Mittal, Eds), VSP, Utrecht, 1994;3
Lukinmaa PL, Mackie E, Thesleff I, Immunohistochemical localization of the matrix glycoproteins – tenascin and the ED – sequence containing form of cellular fibronectin –in human permanent teeth and periodontal ligament, Journal of Dental Research, 1991; 70: 19-26
Martinez-Hernandez A, Amenta PS, The basement membrane in pathology, Laboratory Investigation, 1983; 48: 656-77
Mata A, Boehm C, Fleischman A, Muschler GF, Roy S, Growth of connective tissue progenitor cells on microtextured polydimethyl siloxane surfaces, J Biomed Mater Res, 2002; 62: 499- 506
Matsuzaka K, Walboomers X F, Yoshinari Masao, Inoue T, Jansen John A, The attachment and growth behavior of osteoblast-like cells on microtextured surfaces, Biomaterials, 2003; 24: 2711-2719
McDonald JC, Whitesides GM, Poly(dimethylsiloxane) for fabricating microfluidic devices, Acc. Chem. Res. , 2002; 35: 491 – 499
Meyle J, Gultig K, Wolburg H, and von Recum A F., Fibroblast anchorage to microtextured surfaces, Journal of Biomedical Research, 1993; 27: 1553-1557
Meyle J, Gultig K, Contact guidance of fibroblasts on biomaterial surfaces, Journal of Materials Science: Materials in Medicine, 1994; 5: 463-466
Michel B, Bernard A, Bietsch, A, Delamarche, Geissler M, Juncker, D, Kind H, Renault J-P, Rothuizen H, Schmid H, Schmidt-Winkel P, StutzR, Wolf H, Printing meets lithography: Soft approaches to high-resolution patterning, IBM J. RES.& DEV, September 2001; 45: No.5
Nagayama K, Matsumoto T, Contribution of actin filaments and microtubules to quasi-in situ tensile properties and internal force balance of cultured smooth muscle cells on a substrate, American Journal of Physiology - Cell Physiology, 2008; 295: (6), pp. C1569-C1578
Narayanan AS, Page RC, Connective tissue of the periodontium: a summary of a current work, Collagen and Related Research, 1983; 3: 33 -64
Naruse K, Application of soft lithography to mechanobiology , Proceedings of the 2005 International Symposium on Micro-NanoMechatronics and Human Science, Eighth Symposium on Micro- and Nano-Mechatronics for Information-Based Society - The 21st Century COE Progr, 2005; art. no. 1589998, 245-250
Oakley C, Brunette DM, The sequence of alignment of microtubules, focal contacts and actin filaments in fibroblasts spreading on smooth and grooved titanium substrata, Journal of Cell Science, 1993; 106: 343-354
Owen MJ, Smith PJ, J. Adhesion Sci. Technol., 1994; 8: 1063
Pecheva E, Pramatarova L, Altankov G, Hydroxyapatite grown on a native extracellular matrix: initial interactions with human fibroblasts, Langmuir, 2007; 23: 9386-9392
Puelo DA, Holleran LA, Doremus RH, Bizios R, Osteoblasts response to orthopedic implant materials in vitro, J Biomed Mater Res., 1991; 25: 711-723
Rogers John A, Nuzzo Ralph G, Recent progress in soft lithography, materialstoday, Februari 2005
Rosso F, Giordano A, Barbarisi M, Barbarisi A, From cell-ECM interactions to tissue engineering, Journal of Cellular Physiology, 2004; 199: 174-180
Rovensky YA, Slavnaja IL, Vasiliev JM, Behavior of Fibroblast-Like Cells on Grooved Surfaces, Exp. Cell Res., 1971; 65: 193-201
Rovensky YA, Slavnaja IL, Spreading of fibroblast-like cells on grooved substrates, Exp. Cell Res., 1974; 84: 199-206
Ruoslahti E, Fibronectin, Journal of Oral Pathology, 1981; 10: 3-13
Scacchi M, The development of the ITI DENTAL IMPLANT SYSTEM, Clinical Oral Implant Research, 2000; 11 (Suppl.):8-11
Scarano A, Piattelli M, Vrespa G., Caputi S, Piattelli A, Bacterial adhesion on titanium nitride-coated and uncoated implants: an in vivo human study, Journal of Oral Implantology, 2003; 29: 80-85
Shuttleworth CA, Berry L, Wilson NHF, Collagen synthesis in rabbit dental pulp fibroblasts cultures, Archives of Oral Biology, 1980; 25: 201-5
Sinha RK, Tuan RS, Regulation of human osteoblast integrin expression by orthopedic implant materials, Bone, 1996; 18: 451-457
Stein GS, Lian J, Stein JL, vanWijnen AJ, Frenkel B, Montecino M, Mechanisms regulating osteoblast proliferation and differentiation. In: Bilezikian JP, Raisz LG, & Rodan GA (eds), Principles of bone biology, Academic Press, San Diego, 1996, 69-86
Suzuki M., Tamada Y, Iwata H, Ikada Y, in Physicochemical Aspects of Polymer Surfaces, KL Mittel, Ed., Plenum, New York, 1983; 2: 923
Taipale J, Keski-Oja J, Growth factors in the extracellular matrix, FASEB J, 1997; 11: 51-59
Thanawala SK, Chaudhury MK, Langmuir, 2000; 16: 1256 and references therein
Van Amerongan Jp, Lemmens IG, Tonino GJM, The concentration, extractability, and characterization of collagen in human dental pulp, Archives of Oral Biology, 1983; 28: 339- 45
Van Kooten TG, Whitesides JF, von Recum AF, Influence of silicone (PDMS) surface texture on human skin fibroblast proliferation as determined by cell cycle analysis, J. Biomed. Mater. Res, 1998; 43: 1-14
Vasilets VN, Nakamura K, Uyama Y, Ogata S, Ikada Y, Polymer, 1998; 39:2875
Von Recum AF, Shannon CE, Cannon EC, Long KJ, Van Kooten TG, Meyle J, Surface roughness, porosity, and texture as modifiers of cellular adhesion, Tissue Engineering, 1996; 2: 241- 253
von Rossum A G S H, Moolenaar WH, Schuuring E, Cortactin affects cell migration by regulating intercellular adhesion and cell spreading, Exp. Cell Res., 2006; 312: 1658-1670
Walboomers XF, Croes HJE, Ginsel LA, Jansen JA, Growth behavior of fibroblasts on microgrooved polystyrene, Biomaterials, 1998; 19: 1861-1868
Walboomers XF, Croes HJE, Ginsel LA, Jansen JA, Microgrooved subcutaneous implants in the goat, J. Biomed. Mater Res., 1998; 42: 634-641
Walboomers XF, Monaghan W, Curtis, ASG, Jansen, JA, Attachment of fibroblasts on smooth and microgrooved polystyrene, Journal of Biomedical Materials Research, 1999; 46 (2): 212-220
Weiss P, Experiments on Cell and Axon Orientation in vitro: The Role of Colloidal Exudates in Tissue Organization, J. Exp. Zool, 1945; 100: 353-386
Whitesides G.M, Xia Y, Angew. Chem. Int. Ed, 1998; 37(5): 550
Whitesides G.M, Stroock AD, Gitlin G.I., Ng JMK, Components for integrated poly (dimethylsiloxane) microfluidic systems, Electrophoresis, 2002; 23: 3461-3473
Yamada KM, Olden K, Fibronectins- adhesive glycoproteins of cell surface and blood, Nature, 1978; 275: 179-84
Yamada Kenneth M, Akiyama Steven K, Hayashi Masao, The structure of fibronectin and its role in cellular adhesion, Journal of Supramolecular Structure and Cellular Biochemistry, 1981; 16: 345-358
Zimerman B, Volberg T, Geiger B, Early molecular events in the assembly of the focal adhesion-stress fiber complex during fibroblast spreading, Cell Motil. Cytoskelet, 2004; 58:143–159