Molly M Stevens

Imperial College London, Londinium, England, United Kingdom

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Publications (222)

  • Eunjung Kim · Philip D. Howes · Spencer W. Crowder · Molly M. Stevens
    [Show abstract] [Hide abstract] ABSTRACT: Combining technological developments such as nanomaterials, DNA nanotechnology and functional enzymes has great potential to facilitate next generation high performance molecular diagnostic systems. In this work we describe a microRNA (miRNA) detection assay that combines target recycling and isothermal amplification in an elegantly designed enzyme-mediated cascade reaction. Target recycling is driven by the action of duplex-specific nuclease (DSN), resulting in highly amplified translation of input miRNA to short output DNA fragments. These fragments act as highly specific initiators of rolling circle amplification (RCA), an isothermal reaction that outputs a large volume of polymeric DNAzymes per initiator, and finally a fluorogenic output signal. Based on careful electrophoretic analysis we observed that the DSN produces ca. 10 nt DNA fragments from DNA/miRNA duplexes, regardless of the length of DNA strands. Target recycling yielded ca. five orders of magnitude amplification through DSN-assisted recycling system on magnetic particles, and the RCA a further two orders of magnitude. The final assay exhibited a limit of detection of 1.8 fM of miRNA spiked into 20% human serum, and showed excellent selectivity for miR-21 versus single base-mismatched sequences and other cancer-related miRNAs. The developed assay was further employed to determine accurate amounts of miR-21 in total RNA samples extracted from human cancer cell lines and normal cells, confirming the applicability of the assay for direct and absolute quantification of mature specific miRNA in real biological samples.
    Article · Oct 2016
  • Damia Mawad · Arbel Artzy-Schnirman · Joanne Tonkin · [...] · Molly M. Stevens
    Dataset · Sep 2016
  • [Show abstract] [Hide abstract] ABSTRACT: Due to their outstanding mechanical properties and excellent biocompatibility, zirconia-toughened alumina (ZTA) ceramics have become the gold standard in orthopedics for the fabrication of ceramic bearing components over the last decade. However, ZTA is bioinert, which hampers its implantation in direct contact with bone. Furthermore, periprosthetic joint infections are now the leading cause of failure for joint arthroplasty prostheses. To address both issues, an improved surface design is required: a controlled micro- and nano-roughness can promote osseointegration and limit bacterial adhesion whereas surface porosity allows loading and delivery of antibacterial compounds. In this work, we developed an integrated strategy aiming to provide both osseointegrative and antibacterial properties to ZTA surfaces. The micro-topography was controlled by injection molding. Meanwhile a novel process involving the selective dissolution of zirconia (selective etching) was used to produce nano-roughness and interconnected nanoporosity. Potential utilization of the porosity for loading and delivery of antibiotic molecules was demonstrated, and the impact of selective etching on mechanical properties and hydrothermal stability was shown to be limited. The combination of injection molding and selective etching thus appears promising for fabricating a new generation of ZTA components implantable in direct contact with bone.
    Article · Sep 2016 · Acta biomaterialia
  • [Show abstract] [Hide abstract] ABSTRACT: Bioglass® was the first synthetic material capable of bonding with bone without fibrous encapsulation, and fulfils some of the criteria of an ideal synthetic bone graft. However, it is brittle and toughness is required. Here, we investigated hybrids consisting of co-networks of high cross- linking density polymethacrylate and silica (class II hybrid) as a potential new generation of scaffold materials. Poly(3-(methoxysilyl)propyl methacrylate) (pTMSPMA) and tetraethyl orthosilicate (TEOS) were used as sol-gel precursors and hybrids were synthesised with different inorganic to organic ratios (Ih). The hybrids were nanoporous, with a modal pore diameter of 1 nm. At Ih = 50 %, the release of silica was controlled by varying the molecular weight of pTMSPMA while retaining a specific surface area above 100 m2 g−1. Strain to failure increased to 14.2 %, for Ih = 50 % using a polymer of 30 kDa, compared to 4.5 % for pure glass. The modulus of toughness (UT ) increased from 0.73 (pure glass) to 2.64 GPa. Although, the hybrid synthesised in this report did not contain calcium, pTMSPMA/SiO2 hybrid was found to nucleate bone-like mineral on its surface after 1 week of immersion in simulated body fluid (SBF), whereas pure silica sol-gel glass did not. This increase in apatite forming ability was due to the ion-dipole complexation of calcium with the ester moieties of the polymer that were exposed after release of soluble silica from TEOS. No adverse cytotoxicity for MC3T3-E1 osteoblast-like cells was detected and improved cell attach- ment was observed, compared to a pure silica gel. pTMSPMA/SiO2 hybrids have potential for the regeneration of hard tissue as they overcome the major drawbacks of pure inorganic substrates while retaining cell attachment.
    Article · Aug 2016 · Journal of Materials Chemistry B
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    Paresh A. Parmar · Stacey C. Skaalure · Lesley W. Chow · [...] · Molly M. Stevens
    File available · Dataset · Aug 2016
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    Justin J. Chung · Siwei Li · Molly M. Stevens · [...] · Julian R. Jones
    [Show abstract] [Hide abstract] ABSTRACT: Bioglass® was the first synthetic biomaterial that formed a chemical bond to bone. Although bioactive glass scaffolds can mimic bone’s porous structure, they are brittle. Sol-gel derived hybrids could overcome this problem because their nanoscale co-networks of silica and organic polymer have potential to provide unique physical properties and con-trolled homogenous biodegradation. Copolymers of methyl methacrylate (MMA) and 3-(trimethoxysilyl)propyl meth-acrylate (TMSPMA) has been used as an organic source for hybrids to take advantage of its self-hardening property. However, the effect of well-defined poly(MMA-co-TMSPMA) architecture in the hybrid system has not been investi-gated. Here, linear, randomly branched and star shaped methacrylate based copolymers were synthesized via reversible addition-fragmentation chain transfer (RAFT) polymerization method. These copolymers were then used to fabricate hybrids. The 3-D polymer structure had a significant effect on mechanical properties, providing higher strain to failure while maintaining a compressive strength similar to sol-gel glass. Star copolymer-SiO2 hybrids had a modulus of toughness 9.6 fold greater, and Young’s modulus 4.5 fold lower than a sol-gel derived bioactive glass. During in vitro cell culture, MC3T3-E1 osteoblast precursor cells adhered on the surface regardless of the polymer structure. Introduc-ing star polymer to inorganic-organic hybrids opens possibilities for fine-tuning physical properties for bone scaffold material.
    Full-text available · Article · Aug 2016 · Chemistry of Materials
  • Article · Aug 2016 · Journal of Biomedical Materials Research Part A
  • Damia Mawad · Arbel Artzy-Schnirman · Joanne Tonkin · [...] · Molly M. Stevens
    [Show abstract] [Hide abstract] ABSTRACT: Poly(ethylene dioxythiophene) with functional pendant groups bearing double bonds is synthesized and employed for the fabrication of electroactive hydrogels with advantageous characteristics: covalently cross-linked porous 3D scaffolds with notable swelling ratio, appropriate mechanical properties, electroactivity in physiological conditions, and suitability for proliferation and differentiation of C2C12 cells. This is a new approach for the fabrication of conductive engineered constructs.
    Article · Jul 2016 · Chemistry of Materials
  • [Show abstract] [Hide abstract] ABSTRACT: A direct observation and an in-depth characterization of the steps by which bone mineral nucleates and grows in the extracellular matrix during the earliest stages of maturation, using relevant biomineralization models as they grow into mature bone mineral, is an important research goal. To better understand the process of bone mineralization in the extracellular matrix, we used nano-analytical electron microscopy techniques to examine an in vitro model of bone formation. This study demonstrates the presence of three dominant CaP structures in the mineralizing osteoblast cultures: <80 nm dense granules with a low calcium to phosphate ratio (Ca/P) and crystalline domains; calcium phosphate needles emanating from a foci: "needle-like globules" (100-300 nm in diameter); and mature mineral, both with statistically higher Ca/P compared to that of the dense granules. Many of the submicron granules and globules were interspersed around fibrillar structures containing nitrogen, which are most likely the signature of the organic phase. With high spatial resolution electron energy loss spectroscopy (EELS) mapping, spatially resolved maps were acquired showing the distribution of carbonate within each mineral structure. The carbonate was located in the middle of the granules, which suggested the nucleation of the younger mineral starts with a carbonate-containing precursor and that this precursor may act as seed for growth into larger, submicron-sized, needle-like globules of hydroxyapatite with a different stoichiometry. Application of analytical electron microscopy has important implications in deciphering both how normal bone forms and in understanding pathological mineralization.
    Article · Jul 2016 · ACS Nano
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    [Show abstract] [Hide abstract] ABSTRACT: Abstract This work reports the synthesis of lithium-silicate glass, containing 10 mol% of Li\(_2\)O by the sol–gel process, intended for the regeneration of cartilage. Lithium citrate and lithium nitrate were selected as lithium precursors. The effects of the lithium precursor on the sol–gel process, and the resulting glass structure, morphology, dissolution behaviour, chondrocyte viability and proliferation, were investigated. When lithium citrate was used, mesoporous glass containing lithium as a network modifier was obtained, whereas the use of lithium nitrate produced relatively dense glass-ceramic with the presence of lithium metasilicate, as shown by X-ray diffraction, \(^{29}\)Si and \(^7\)Li MAS NMR and nitrogen sorption data. Nitrate has a better affinity for lithium than citrate, leading to heterogeneous crystallisation from the mesopores, where lithium salts precipitated during drying. Citrate decomposed at a lower temperature, where the crystallisation of lithium-silicate crystal is not thermodynamically favourable. Upon decomposition of the citrate, a solid-state salt metathesis reaction between citrate and silanol occurred, followed by the diffusion of lithium within the structure of the glass. Both glass and glass-ceramic released silica and lithium ions in culture media, but release rate was lower for the glass-ceramic. Both samples did not affect chondrocyte viability and proliferation. Graphical Abstract
    Full-text available · Article · Jun 2016 · Journal of Sol-Gel Science and Technology
  • Nathan J. Liu · Robert Chapman · Yiyang Lin · [...] · Molly M. Stevens
    [Show abstract] [Hide abstract] ABSTRACT: Acute pancreatitis is a relatively common and potentially fatal condition, but the presenting symptoms are non-specific and diagnosis relies largely on the measurement of amylase activity by the hospital clinical laboratory. In this work we develop a point of care test for pancreatitis measuring concentration of secretory phospholipase A2 group IB (sPLA2-IB). Novel antibodies for sPLA2-IB were raised and used to design an ELISA and a lateral flow device (LFD) for the point of care measurement of sPLA2-IB concentration, which was compared to pancreatic amylase activity, lipase activity, and sPLA2-IB activity in 153 serum samples. 98 of these samples were obtained from the pathology unit of a major hospital and classified retrospectively according to presence or absence of pancreatitis, and the remaining 55 were obtained from commercial sources to serve as high lipase (n = 20), CA19-9 positive (n = 15), and healthy (n = 20) controls. sPLA2-IB concentration correlated well with the serum activity of both amylase and lipase, and performed at least as well as either markers in the differentiation of pancreatitis from controls.
    Article · May 2016 · Nanoscale
  • Paresh A. Parmar · Jean-Philippe St-Pierre · Lesley W. Chow · [...] · Molly M. Stevens
    [Show abstract] [Hide abstract] ABSTRACT: Collagen I foams are used in the clinic as scaffolds to promote articular cartilage repair as they provide a bioactive environment for cells with chondrogenic potential. However, collagen I as a base material does not allow for precise control over bioactivity. Alternatively, recombinant bacterial collagens can be used as "blank slate" collagen molecules to offer a versatile platform for incorporation of selected bioactive sequences and fabrication into 3D scaffolds. Here, we show the potential of Streptococcal collagen-like 2 (Scl2) protein foams modified with peptides designed to specifically and noncovalently bind hyaluronic acid and chondroitin sulfate to improve chondrogenesis of human mesenchymal stem cells (hMSCs) compared to collagen I foams. Specific compositions of functionalized Scl2 foams lead to improved chondrogenesis compared to both nonfunctionalized Scl2 and collagen I foams, as indicated by gene expression, extracellular matrix accumulation, and compression moduli. hMSCs cultured in functionalized Scl2 foams exhibit decreased collagens I and X gene and protein expression, suggesting an advantage over collagen I foams in promoting a chondrocytic phenotype. These highly modular foams can be further modified to improve specific aspects chondrogenesis. As such, these scaffolds also have the potential to be tailored for other regenerative medicine applications.
    Article · May 2016 · Advanced Healthcare Materials
  • Paresh A. Parmar · Stacey C. Skaalure · Lesley W. Chow · [...] · Molly M. Stevens
    [Show abstract] [Hide abstract] ABSTRACT: Tissue engineering strategies for repairing and regenerating articular cartilage face critical challenges to recapitulate the dynamic and complex biochemical microenvironment of native tissues. One approach to mimic the biochemical complexity of articular cartilage is through the use of recombinant bacterial collagens as they provide a well–defined biological ‘blank template’ that can be modified to incorporate bioactive and biodegradable peptide sequences within a precisely defined three–dimensional system. We customized the backbone of a Streptococcal collagen–like 2 (Scl2) protein with heparin–binding, integrin–binding, and hyaluronic acid–binding peptide sequences previously shown to modulate chondrogenesis and then cross–linked the recombinant Scl2 protein with a combination of matrix metalloproteinase 7 (MMP7)– and aggrecanase (ADAMTS4)–cleavable peptides at varying ratios to form biodegradable hydrogels with degradation characteristics matching the temporal expression pattern of these enzymes in human mesenchymal stem cells (hMSCs) during chondrogenesis. hMSCs encapsulated within the hydrogels cross–linked with both degradable peptides exhibited enhanced chondrogenic characteristics as demonstrated by gene expression and extracellular matrix deposition compared to the hydrogels cross–linked with a single peptide. Additionally, these combined peptide hydrogels displayed increased MMP7 and ADAMTS4 activities and yet increased compression moduli after 6 weeks, suggesting a positive correlation between the degradation of the hydrogels and the accumulation of matrix by hMSCs undergoing chondrogenesis. Our results suggest that including dual degradation motifs designed to respond to enzymatic activity of hMSCs going through chondrogenic differentiation led to improvements in chondrogenesis. Our hydrogel system demonstrates a bimodal enzymatically degradable biological platform that can mimic native cellular processes in a temporal manner. As such, this novel collagen–mimetic protein, cross–linked via multiple enzymatically degradable peptides, provides a highly adaptable and well defined platform to recapitulate a high degree of biological complexity, which could be applicable to numerous tissue engineering and regenerative medicine applications.
    Article · May 2016 · Biomaterials
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    Andrew Shevchuk · Sergiy Tokar · Sahana Gopal · [...] · Yuri E. Korchev
    [Show abstract] [Hide abstract] ABSTRACT: Scanning ion conductance microscopy (SICM) is a super-resolution live imaging technique that uses a glass nanopipette as an imaging probe to produce three-dimensional (3D) images of cell surface. SICM can be used to analyze cell morphology at nanoscale, follow membrane dynamics, precisely position an imaging nanopipette close to a structure of interest, and use it to obtain ion channel recordings or locally apply stimuli or drugs. Practical implementations of these SICM advantages, however, are often complicated due to the limitations of currently available SICM systems that inherited their design from other scanning probe microscopes in which the scan assembly is placed right above the specimen. Such arrangement makes the setting of optimal illumination necessary for phase contrast or the use of high magnification upright optics difficult. Here, we describe the designs that allow mounting SICM scan head on a standard patch-clamp micromanipulator and imaging the sample at an adjustable approach angle. This angle could be as shallow as the approach angle of a patch-clamp pipette between a water immersion objective and the specimen. Using this angular approach SICM, we obtained topographical images of cells grown on nontransparent nanoneedle arrays, of islets of Langerhans, and of hippocampal neurons under upright optical microscope. We also imaged previously inaccessible areas of cells such as the side surfaces of the hair cell stereocilia and the intercalated disks of isolated cardiac myocytes, and performed targeted patch-clamp recordings from the latter. Thus, our new, to our knowledge, angular approach SICM allows imaging of living cells on nontransparent substrates and a seamless integration with most patch-clamp setups on either inverted or upright microscopes, which would facilitate research in cell biophysics and physiology.
    Full-text available · Article · May 2016 · Biophysical Journal
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    Nadav Amdursky · Xuhua Wang · Paul Meredith · [...] · Molly M. Stevens
    [Show abstract] [Hide abstract] ABSTRACT: Supplementary
    File available · Dataset · Apr 2016
  • Charalambos Kallepitis · Helene Autefage · Felix Allen · [...] · Molly M. Stevens
    Conference Paper · Mar 2016
  • Robert Chapman · Adam J Gormley · Martina H Stenzel · Molly M Stevens
    [Show abstract] [Hide abstract] ABSTRACT: The synthesis of well-defined polymers in a low-volume, combinatorial fashion has long been a goal in polymer chemistry. Here, we report the preparation of a wide range of highly controlled homo and block co-polymers by Enz-RAFT (enzyme-assisted reversible addition-fragmentation chain transfer) polymerization in microtiter plates in the open atmosphere. The addition of 1 μm glucose oxidase (GOx) to water/solvent mixtures enables polymerization reactions to proceed in extremely low volumes (40 μL) and low radical concentrations. This procedure provides excellent control and high conversions across a range of monomer families and molecular weights, thus avoiding the need to purify for screening applications. This simple technique enables combinatorial polymer synthesis in microtiter plates on the benchtop without the need of highly specialized synthesizers and at much lower volumes than is currently possible by any other technique.
    Article · Mar 2016 · Angewandte Chemie International Edition
  • Martin A. B. Hedegaard · Mads S. Bergholt · Molly M. Stevens
    [Show abstract] [Hide abstract] ABSTRACT: Imaging by Raman spectroscopy enables unparalleled label-free insights into cell and tissue composition at the molecular level. With established approaches limited to single image analysis, there are currently no general guidelines or consensus on how to quantify biochemical components across multiple Raman images. Here, we describe a broadly applicable methodology for the combination of multiple Raman images into a single image for analysis. This is achieved by removing image specific background interference, unfolding the series of Raman images into a single dataset, and normalisation of each Raman spectrum to render comparable Raman images. Multivariate image analysis is finally applied to derive the contributing 'pure' biochemical spectra for relative quantification. We present our methodology using four independently measured Raman images of control cells and four images of cells treated with strontium ions from substituted bioactive glass. We show that the relative biochemical distribution per area of the cells can be quantified. In addition, using k-means clustering, we are able to discriminate between the two cell types over multiple Raman images. This study shows a streamlined quantitative multi-image analysis tool for improving cell/tissue characterisation and opens new avenues in biomedical Raman spectroscopic imaging.
    Article · Mar 2016 · Journal of Biophotonics
  • [Show abstract] [Hide abstract] ABSTRACT: The increasing demand for early detection of diseases drives the efforts to develop more and more sensitive techniques to detect biomarkers in extremely low concentrations. Electromagnetic modes at the surface of one dimensional photonic crystals, usually called Bloch surface waves, were demonstrated to enhance the resolution and constitute an attractive alternative to surface plasmon polariton optical biosensors. We report on the development of Bloch surface wave biochips operating in both label-free and fluorescence modes and demonstrate their use in ovalbumin recognition assays.
    Article · Mar 2016 · Proceedings of SPIE - The International Society for Optical Engineering
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    File available · Dataset · Feb 2016

Publication Stats

9k Citations


  • 1970-2015
    • Imperial College London
      • • Department of Materials
      • • Institute of Biomedical Engineering (IBME)
      Londinium, England, United Kingdom