W. Richard Bowen and Nidal Hilal 4

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218 7. MICRO/NANOENgINEERINg ANd AFM FOR CELLULAR SENSINg or physical in nature, have been reliably produced by various methods. Many of these structures have emerged from the novel combination of advances in micro- or nanofabrication, chemistry and biology. Surface patterning methods offer considerable flexibility in pattern design, ligand specificity and density, and surface composition to investigate interactions of cells with chemical variations in the ECM. A vast range of engineered topographic features, from micro- to nanoscale, have been found to play a significant regulatory role in cellular proliferation, migration and differentiation. These studies provide significant insights into mechanistic questions of how cells are able to sense, integrate and respond to collective chemical and mechanical signals. In addition, they also demonstrate that engineered environments can regulate cell functions and fate, and thus open new opportunities of developing tailored biosubstrates for specific tissue cell types, although this is still at its early stage. Optical microscopes and SEM have been essential tools for many of these studies; however, they are subject to many constraints in exploring the molecular mechanisms at subcellular or cellular level, which are essential for the control of the biological pathway. AFM, having been established as an important tool in development of nanopatterned surfaces, has provided increased momentum to living system studies. Nanometre spatial imaging and quantitative measurement of mechanical properties of functional components of a living cell have been reliably achieved. Long standing questions in the nanoworld, such as whether it is nanoscale topographic features or chemical patterns that predominate in inducing cellular reactions [40], may be answerable. The combination of AFM with existing optical techniques has already shown itself to be a powerful tool, and further insights will be gained into in vitro cell differentiation and disease diagnostics. Together with the engineered environments that can be employed to guide cell function and fate, it might be possible to move towards controlling biological pathways in cell culture, and so explore new medicines and therapies for human health. There are still many open challenges in cell biology and in physiological and pathological dysfunctions. The convergence of micro- or nanoengineering, AFM and interfacial chemistry with cell biology tools might provide new opportunities to address these challenges. For example, a better understanding of the complex processes associated with a whole intact cell interacting with individual molecules when cells are in contact with a surface will facilitate the diagnostics of arthoroplastic failures and improve existing implants. A more ambitious approach is to regenerate failed tissue outside a body; this requires an engineered ECM with the functionalities that are found in a body. Although there have been intensive efforts in these fields, researchers are still a long way from recreating an ECM with similar molecular architecture and functionalities of the ECM found in vivo.
AbbREvIATIONS ANd SyMbOLS 219 Increasing collaborations between scientists and communities from various disciplines, including engineering, nanobiotechnology, materials, biology and clinical medicine, are essential to approach this goal. ACknowlEdgEMEntS H. Yin is supported by the Royal Society of Edinburgh as an RSE personal research fellow. G. McPhee thanks the EPSRC for his studentship. P. S. Dobson is supported by an RCUK Academic Research Fellowship. We would like to thank Dr Matthew Dalby for proof reading and offering constructive suggestions and samples. We also thank Drs Andrew Glide and Mathis Riehle and Professor Jon Cooper for their support and discussions. AbbrEvIAtIonS And SyMbolS � Semi-opening angle of the tip ° AFM Atomic force microscope DNA Deoxyribonucleic acid, a nucleic acid E Young’s modulus N m�2 EBL Electron beam lithography ECM Extracellular matrix F Applied force N MSCs Human mesenchymal stem cells NIL Nanoimprinting lithography PBrS Poly(4-bromostyrene) PDMS Poly(dimethysiloxne) PEG Poly(ethylene glycol) PMMA Poly(methyl methacrylate) PS Polystyrene RGD A peptide sequence consisting of Arg-Gly-Asp SAM Self-assembly monolayer SEM Scanning electron microscopy TEM Transmission electron microscopy V Poisson’s ratio, dimensionless � Indentation depth m
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Butterworth-Heinemann is an imprint
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x Preface in their work. Hence, it
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xii Preface them from hostile condi
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xiv About thE Editors Professor Nid
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xvi List of Contributors Dr Daniel
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2 1. BAsIC PRINCIPLEs OF ATOMIC FOR
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20 1. BAsIC PRINCIPLEs OF ATOMIC FO
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32 2. MEASUREMENT OF PARTICLE ANd S
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82 3. QUANTIFICATION OF PARTICLE-BU
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100 3. QUANTIFICATION OF PARTICLE-B
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C H A P T E R 4 Investigating Membr
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4.2 THE RANgE OF POssIbILITIEs FOR
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4.2 THE RANgE OF POssIbILITIEs FOR
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4.3 CORREsPONdENCE bETwEEN sURFACE
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4.3 CORREsPONdENCE bETwEEN sURFACE
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4.4 IMAgINg IN LIqUId ANd THE dETER
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4.4 IMAgINg IN LIqUId ANd THE dETER
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4.5 EFFECTs OF sURFACE ROUgHNEss ON
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4.6 ‘vIsUALIsATION’ OF THE REjE
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4.7 THE UsE OF AFM IN MEMbRANE dEvE
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Dfractional 0.18 0.16 0.14 0.12 0.1
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4.8 CHARACTERIsATION OF METAL sURFA
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At pH 5.5, dissolution began to occ
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REFERENCEs 137 [4] W.R. Bowen, N. H
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C H A P T E R 5 AFM and Development
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5.2 MEAsUREMENT OF ADHEsION OF COLL
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5.2 MEAsUREMENT OF ADHEsION OF COLL
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The antibacterial activity of initi
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5.3 MODIFICATION OF MEMBRANEs 147 t
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5.3 MODIFICATION OF MEMBRANEs 149 B
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Force (nN m -1 ) Loading force (mN
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5.3 MODIFICATION OF MEMBRANEs 153 p
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Adhesion force (mN m -1 ) 10 8 6 4
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Adhesion force (mN m -1 ) 20 15 10
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Adhesion force (mN m -1 ) 10 8 6 4
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5.3 MODIFICATION OF MEMBRANEs 161 F
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5.4 MODIFICATION OF MEMBRANEs wITH
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5.4 MODIFICATION OF MEMBRANEs wITH
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268 9. APPLICATION OF ATOMIC FORCE
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276 10. FuTuRE PRosPECTs most AFM s
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278 10. FuTuRE PRosPECTs targeted d
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A Actin stress fiber, 197 Adhesion,
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Natural organic matter, 140 Negativ
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W. Richard Bowen and Nidal Hilal 4

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