Cytoskeleton remodelling of confluent epithelial cells cultured on porous substrates

The impact of substrate topography on the morphological and mechanical properties of confluent MDCK-II cells cultured on porous substrates was scrutinized by means of various imaging techniques as well as atomic force microscopy comprising force volume and microrheology measurements. Regardless of the pore size, ranging from 450 to 5500 nm in diameter, cells were able to span the pores. They did not crawl into the holes or grow around the pores. Generally, we found that cells cultured on non-porous surfaces are stiffer, i.e. cortical tension rises from 0.1 to 0.3 mN m⁻¹, and less fluid than cells grown over pores. The mechanical data are corroborated by electron microscopy imaging showing more cytoskeletal filaments on flat samples in comparison to porous ones. By contrast, cellular compliance increases with pore size and cells display a more fluid-like behaviour on larger pores. Interestingly, cells on pores larger than 3500 nm produce thick actin bundles that bridge the pores and thereby strengthen the contact zone of the cells.     

Insect adhesion on rough surfaces: analysis of adhesive contact of smooth and hairy pads on transparent microstructured substrates

Insect climbing footpads are able to adhere to rough surfaces, but the details of this capability are still unclear. To overcome experimental limitations of randomly rough, opaque surfaces, we fabricated transparent test substrates containing square arrays of 1.4 µm diameter pillars, with variable height (0.5 and 1.4 µm) and spacing (from 3 to 22 µm). Smooth pads of cockroaches (Nauphoeta cinerea) made partial contact (limited to the tops of the structures) for the two densest arrays of tall pillars, but full contact (touching the substrate in between pillars) for larger spacings. The transition from partial to full contact was accompanied by a sharp increase in shear forces. Tests on hairy pads of dock beetles (Gastrophysa viridula) showed that setae adhered between pillars for larger spacings, but pads were equally unable to make full contact on the densest arrays. The beetles'' shear forces similarly decreased for denser arrays, but also for short pillars and with a more gradual transition. These observations can be explained by simple contact models derived for soft uniform materials (smooth pads) or thin flat plates (hairy-pad spatulae). Our results show that microstructured substrates are powerful tools to reveal adaptations of natural adhesives for rough surfaces.     

Human fibroblast attachment on fibrous parylene-C thin-film substrates

Although the polymeric form of parylene-C is used in many medical devices, the mechanistic nature of cellular attachment to polymeric parylene-C is not clear. We examined the effects of (i) substrate morphology, (ii) surface wettability and (iii) presence of serum proteins on fibroblast attachment. A physicochemical vapor deposition technique was implemented to deposit flat parylene-C substrates as well as fibrous substrates of three different morphologies: slanted columnar, chevronic and chiral. Flat parylene-C surfaces were moderately hydrophobic while fibrous substrates were superhydrophobic. Pretreatment with oxygen plasma changed the substrate surfaces from hydrophobic to superhydrophilic. The attachment efficiency of human fibroblast cells to the flat and three fibrous thin-film parylene-C substrates was investigated. Fibroblast attachment was better on fibrous substrates than on flat substrates, and oxygen plasma pretreatment facilitated fibroblast attachment on all four morphologies. Serum proteins also facilitated cell attachment on all substrates. The combination of oxygen plasma pre-treatment and serum proteins increased fibroblast adhesion in an additive manner on flat, but not on fibrous parylene-C substrates. The morphology of cell–substrate interactions differed between fibrous and flat parylene-C substrates.     

中温固体氧化物燃料电池阳极基底及负载型电解质薄膜的研制

在用固相反应法合成电解质材料Ｌａ_０．９Ｓｒ_０．１Ｇａ_０．８Ｍｇ_０．２Ｏ_３－δ（ＬＳＧＭ）的基础上,研制出中温固体氧化物燃料电池ＬＳＧＭ＋ＮｉＯ阳极基底;考察了阳极基底孔隙率、孔径分布及电导率随组成变化的规律;研究了阳极基底组成、微观结构、制备工艺等对负载型ＬＳＧＭ电解质薄膜成膜过程及质量的影响和负载型电解质薄膜在还原气氛中的结构稳定性;采用湿化学物理方法及等静压烧结工艺成功地制备出了厚度为２０～５０μｍ的负载型致密ＬＳＧＭ电解质薄膜．研究表明;ＮｉＯ含量为６０％的阳极基底具有适宜的烧结收缩率、孔隙率与孔径分布,且比表面积与比孔容积均较大,适合作为ＳＯＦＣ的阳极．随着ＮｉＯ含量的增加,还原后阳极基底的电导率有所增大．其中低ＮｉＯ含量的阳极基底在还原后的初生态,其电导率在交变信号的诱导下发生弛豫现象而迅速增大,并由离子导电性转变为金属导电性．而高ＮｉＯ含量的阳极基底,其还原后的电导率随测量时间的延长变化很小,并从一开始就表现出金属导电的性质．采用无约束烧结程序制备的负载型ＬＳＧＭ电解质薄膜,表面为粗大的片状晶粒,还原后在晶界处产生裂纹．而采用等静压烧结程序制备的负载型ＬＳＧＭ电解质薄膜,表面为细小、形状规则的晶粒, 晶界结合紧密, 且还原后晶界无裂纹出现.     

Extracellular-matrix tethering regulates stem-cell fate

To investigate how substrate properties influence stem-cell fate, we cultured single human epidermal stem cells on polydimethylsiloxane (PDMS) and polyacrylamide (PAAm) hydrogel surfaces, 0.1?kPa-2.3?MPa in stiffness, with a covalently attached collagen coating. Cell spreading and differentiation were unaffected by polydimethylsiloxane stiffness. However, cells on polyacrylamide of low elastic modulus (0.5?kPa) could not form stable focal adhesions and differentiated as a result of decreased activation of the extracellular-signal-related kinase (ERK)/mitogen-activated protein kinase (MAPK) signalling pathway. The differentiation of human mesenchymal stem cells was also unaffected by PDMS stiffness but regulated by the elastic modulus of PAAm. Dextran penetration measurements indicated that polyacrylamide substrates of low elastic modulus were more porous than stiff substrates, suggesting that the collagen anchoring points would be further apart. We then changed collagen crosslink concentration and used hydrogel-nanoparticle substrates to vary anchoring distance at constant substrate stiffness. Lower collagen anchoring density resulted in increased differentiation. We conclude that stem cells exert a mechanical force on collagen fibres and gauge the feedback to make cell-fate decisions.     

Optimizing PANi doped electroactive substrates as patches for the regeneration of cardiac muscle

In scaffold aided regeneration of muscular tissue, composite materials are currently utilized as a temporary substrate to stimulate tissue formation by controlled electrochemical signals as well as continuous mechanical stimulation until the regeneration processes are completed. Among them, composites from the blending of conductive (CPs) and biocompatible polymers are powerfully emerging as a successful strategy for the regeneration of myocardium due to their unique conductive and biological recognition properties able to assure a more efficient electroactive stimulation of cells. Here, different composite substrates made of synthesized polyaniline (sPANi) and polycaprolactone (PCL) were investigated as platforms for cardiac tissue regeneration. Preliminary, a comparative analysis of substrates conductivity performed on casted films endowed with synthesized polyaniline (sPANi) short fibres or blended with emeraldine base polyaniline (EBPANi) allows to study the attitude of charge transport, depending on the conducting filler amount, shape and spatial distribution. In particular, conducibility tests indicated that sPANi short fibres provide a more efficient transfer of electric signal due to the spatial organization of electroactive needle-like phases up to form a percolative network. On the basis of this characterization, sPANi/PCL electrospun membranes have been also optimized to mimic either the morphological and functional features of the cardiac muscle ECM. The presence of sPANi does not relevantly affect the fibre architecture as confirmed by SEM/image analysis investigation which shows a broader distribution of fibres with only a slight reduction of the average fibre diameter from 7.1 to 6.4 μm. Meanwhile, biological assays—evaluation of cell survival rate by MTT assay and immunostaining of sarcomeric α-actinin of cardiomyocites-like cells—clearly indicate that conductive signals offered by PANi needles, promote the cardiogenic differentiation of hMSC into cardiomyocite-like cells. These preliminary results concur to promise the development of electroactive biodegradable substrates able to efficiently stimulate the basic cell mechanisms, paving the way towards a new generation of synthetic patches for the support of the regeneration of damaged myocardium.     

Efficient co‐cultivation of human fibroblast cells (HFCs) and adipose‐derived stem cells (ADSs) on gelatin/PLCL nanofiber

In this study, we investigated whether the nanofibers produced by natural‐synthetic polymers can probably promote the proliferation of co‐cultured adipose‐derived stem cells/human fibroblast cells (ADSs/HFCs) and synthesis of collagen. Nanofiber was fabricated by blending gelatin and poly (L‐lactide co‐ɛ‐caprolactone) (PLCL) polymer nanofiber (Gel/PLCL). Cell morphology and the interaction between cells and Gel/PLCL nanofiber were evaluated by FESEM and fluorescent microscopy. MTS assay and quantitative real‐time polymerase chain reaction were applied to assess the proliferation of co‐cultured ADSs/HFCs and the collagen type I and III synthesis, respectively. The concentrations of two cytokines including fibroblast growth factor‐basic and transforming growth factor‐β1 were also measured in culture medium of co‐cultured ADSs/HDCs using enzyme‐linked immunosorbent assay assay. Actually, nanofibers exhibited proper structural properties in terms of stability in cell proliferation and toxicity analysis processes. Gel/PLCL nanofiber promoted the growth and the adhesion of HFCs. Our results showed in contact co‐culture of ADSs/HFCs on the Gel/PLCL nanofiber increased cellular adhesion and proliferation synergistically compared to non‐coated plate. Also, synthesis of collagen and cytokines secretion of co‐cultured ADSs/HFCs on Gel/PLCL scaffolds is significantly higher than non‐coated plates. To conclude, the results suggest that Gel/PLCL nanofiber can imitate physiological characteristics in vivo and enhance the efficacy of co‐cultured ADSs/HFCs in wound healing process.Inspec keywords: biomedical materials, enzymes, adhesion, fluorescence, polymer fibres, tissue engineering, wounds, nanofibres, cellular biophysics, molecular biophysics, gelatin, biochemistry, nanomedicine, field emission scanning electron microscopy, nanofabricationOther keywords: cell morphology, cell proliferation, efficient cocultivation, HFCs, ADSs, gelatin‐PLCL nanofiber, natural‐synthetic polymers, cocultured adipose‐derived stem cells‐human fibroblast cells, FESEM, fluorescent microscopy, MTS assay, quantitative real‐time polymerase chain reaction, collagen type I synthesis, collagen type III synthesis, cytokines, transforming growth factor‐β1, fibroblast growth factor‐basic growth factor‐β1, culture medium, enzyme‐linked immunosorbent assay assay, structural properties, toxicity analysis, cellular adhesion, physiological characteristics in vivo, wound healing     

The effect of nanoscale surface curvature on the oligomerization of surface-bound proteins

The influence of surface topography on protein conformation and association is used routinely in biological cells to orchestrate and coordinate biomolecular events. In the laboratory, controlling the surface curvature at the nanoscale offers new possibilities for manipulating protein–protein interactions and protein function at surfaces. We have studied the effect of surface curvature on the association of two proteins, α-lactalbumin (α-LA) and β-lactoglobulin (β-LG), which perform their function at the oil–water interface in milk emulsions. To control the surface curvature at the nanoscale, we have used a combination of polystyrene (PS) nanoparticles (NPs) and ultrathin PS films to fabricate chemically pure, hydrophobic surfaces that are highly curved and are stable in aqueous buffer. We have used single-molecule force spectroscopy to measure the contour lengths L_c for α-LA and β-LG adsorbed on highly curved PS surfaces (NP diameters of 27 and 50 nm, capped with a 10 nm thick PS film), and we have compared these values in situ with those measured for the same proteins adsorbed onto flat PS surfaces in the same samples. The L_c distributions for β-LG adsorbed onto a flat PS surface contain monomer and dimer peaks at 60 and 120 nm, respectively, while α-LA contains a large monomer peak near 50 nm and a dimer peak at 100 nm, with a tail extending out to 200 nm, corresponding to higher order oligomers, e.g. trimers and tetramers. When β-LG or α-LA is adsorbed onto the most highly curved surfaces, both monomer peaks are shifted to much smaller values of L_c. Furthermore, for β-LG, the dimer peak is strongly suppressed on the highly curved surface, whereas for α-LA the trimer and tetramer tail is suppressed with no significant change in the dimer peak. For both proteins, the number of higher order oligomers is significantly reduced as the curvature of the underlying surface is increased. These results suggest that the surface curvature provides a new method of manipulating protein–protein interactions and controlling the association of adsorbed proteins, with applications to the development of novel biosensors.     

Neuronal networks in vitro: formation and organization on biofunctionalized surfaces

Receptor-mediated recognition of substrate molecules is a prerequisite for nerve cells in order to find their target structures in vivo and leads to formation of neuronal connections and networks. In order to study these mechanisms under in vitro conditions, we cultured embryonic hippocampal neurons or neuronal cell lines, SH-SY5Y and PCC7-PCC7-Mz1, onto biofunctionalized surfaces. Micropatterning on polymer surfaces, glass- and silicone-oxide-based chip materials was performed in a micrometer range by microcontact printing using polydimethylsiloxane (PDMS) stamps. Hippocampal neurons were found to form networks on chip surfaces under serum-free conditions and remained functional for more than a week. Human neuroblastoma cells SH-SY5Y as well as PCC7-Mz1 stem cells were found to follow microcontact printed pattern on polystyrene surfaces. Both cell lines showed neuronal marker expression and were cultured for up to 7 days with serum containing culture medium. Widths of 3–5 µm of coating lines were found to enhance single cell spreading along the pattern. The techniques described in this study may be useful in promoting nerve cell regeneration and organization following transection due to trauma or surgery. The neuronal alignment and network formation in vitro may furthermore serve as a model system in the field of biosensors. © 1999 Kluwer Academic Publishers     

Honey‐incorporated nanofibre reduces replicative senescence of umbilical cord‐derived mesenchymal stem cells

Umbilical cord‐derived mesenchymal stem cells (UCDMSC) are attractive candidates for cell‐based regenerative medicine. However, they are susceptible to replicative senescence during repetitive passaging for in‐vitro expansion and induced senescence in an oxidative, inflammatory microenvironment in vivo. Aim of this study is to investigate if honey‐incorporated matrices can be employed to reduce senescence of UCDMSC. Matrices were prepared by electrospinning solutions of honey with poly‐vinyl alcohol (PVA). PVA:honey matrices exhibited free radical scavenging activity. Culture of UCDMSC on PVA:honey matrices showed improvement in cell proliferation compared to pure PVA nanofibres. Expression of vimentin indicated that mesenchymal phenotype is preserved after culturing on these matrices. Further, UCDMSC were serially subcultured and cells of two passages (P2 and P6) were evaluated for reactive oxygen species (ROS) load and senescence parameters. P6 cells showed a higher ROS load and β‐galactosidase (β‐gal) positive senescent cells compared to P2. However, culturing on PVA:honey substrates significantly reduced both ROS and β‐gal markers compared to cells on PVA substrates. Honey contains several antioxidant and anti‐inflammatory components, which can reduce the ROS‐related senescence. Thus, it is concluded that honey containing nanofibres can be effective substrates for stem cell‐based wound healing and regenerative medicine.Inspec keywords: molecular biophysics, nanofibres, nanomedicine, polymer fibres, cellular biophysics, nanofabrication, enzymes, biochemistry, electrospinning, wounds, biomedical materialsOther keywords: pure PVA nanofibres, UCDMSC, PVA:honey substrates, PVA substrates, ROS‐related senescence, honey containing nanofibres, stem cell‐based wound healing, honey‐incorporated nanofibre, replicative senescence, umbilical cord‐derived mesenchymal stem cells, cell‐based regenerative medicine, induced senescence, PVA:honey matrices, cell proliferation, honey‐incorporated matrices, electrospinning solutions, poly‐vinyl alcohol, free radical scavenging activity, vimentin expression, mesenchymal phenotype, reactive oxygen species load, senescence parameters, P6 cells, β‐galactosidase positive senescent cells, β‐gal markers, antiinflammatory components, antioxidant components     

The interplay of cell–cell and cell–substrate adhesion in collective cell migration

Collective cell migration often involves notable cell–cell and cell–substrate adhesions and highly coordinated motion of touching cells. We focus on the interplay between cell–substrate adhesion and cell–cell adhesion. We show that the loss of cell-surface contact does not significantly alter the dynamic pattern of protrusions and retractions of fast migrating amoeboid cells (Dictyostelium discoideum), but significantly changes their ability to adhere to other cells. Analysis of the dynamics of cell shapes reveals that cells that are adherent to a surface may coordinate their motion with neighbouring cells through protrusion waves that travel across cell–cell contacts. However, while shape waves exist if cells are detached from surfaces, they do not couple cell to cell. In addition, our investigation of actin polymerization indicates that loss of cell-surface adhesion changes actin polymerization at cell–cell contacts. To further investigate cell–cell/cell–substrate interactions, we used optical micromanipulation to form cell–substrate contact at controlled locations. We find that both cell-shape dynamics and cytoskeletal activity respond rapidly to the formation of cell–substrate contact.     

Adhesive interactions of geckos with wet and dry fluoropolymer substrates

Fluorinated substrates like Teflon^® (poly(tetrafluoroethylene); PTFE) are well known for their role in creating non-stick surfaces. We showed previously that even geckos, which can stick to most surfaces under a wide variety of conditions, slip on PTFE. Surprisingly, however, geckos can stick reasonably well to PTFE if it is wet. In an effort to explain this effect, we have turned our attention to the role of substrate surface energy and roughness when shear adhesion occurs in media other than air. In this study, we removed the roughness component inherent to commercially available PTFE and tested geckos on relatively smooth wet and dry fluoropolymer substrates. We found that roughness had very little effect on shear adhesion in air or in water and that the level of fluorination was most important for shear adhesion, particularly in air. Surface energy calculations of the two fluorinated substrates and one control substrate using the Tabor–Winterton approximation and the Young–Dupré equation were used to determine the interfacial energy of the substrates. Using these interfacial energies we estimated the ratio of wet and dry normal adhesion for geckos clinging to the three substrates. Consistent with the results for rough PTFE, our predictions show a qualitative trend in shear adhesion based on fluorination, and the quantitative experimental differences highlight the unusually low shear adhesion of geckos on dry smooth fluorinated substrates, which is not captured by surface energy calculations. Our work has implications for bioinspired design of synthetics that can preferentially stick in water but not in air.     

A neural cell culture study on thin film electrode materials

Functional neural stimulation requires good interface between the neural cells and the electrode surfaces. In order to study the effect of electrode materials and surface structure on cell adhesion and biocompatibility, we cultured cortical neurons on thin films of platinum and iridium oxide. We used both flat, as-deposited and laser micro-structured films. The laser micro-structuring consisted of creating regular arrays of micro-bumps or holes with diameters of 4–5 μm and height of about 1.5 μm. The micro-bumps were fabricated onto platinum and iridium film surfaces deposited on borosilicate glass substrates, using mask-projection irradiation with single nano-second pulses from a KrF excimer laser (λ = 248 nm). Amorphous and crystalline (deposited at 250 °C) IrO₂ films were deposited onto the laser micro-structured iridium films by pulsed-DC reactive sputtering to obtain micro-structured IrO₂ films. Cortical neurons isolated from rat embryo brain were cultured onto these film surfaces. Our results indicate that flat and micro-structured film surfaces are biocompatible and non-toxic for neural cell growth. The use of poly-d-lysine as a mediator for cell adhesion onto the thin film surfaces is also discussed.     

Differentiation of human mesenchymal stem cells on nano- and micro-grain size titania

The current study investigated the osteogenic differentiation of human mesenchymal stem cells cultured on titania surfaces with grain sizes ranging from 50 to 1500 nm in either control or osteogenic medium. Characterization of osteogenic differentiation included quantification of the osteopontin and alkaline phosphatase expression by the cells, as well as of the content of calcium in the extracellular matrix. Mesenchymal stem cell differentiation was not observed on any of the grain sizes tested without dexamethasone and osteogenic-stimulating chemical agents (specifically, ascorbic acid and beta-glycerolphosphate) in the culture medium. Little-to-no mesenchymal stem cell differentiation was detected on the 50 nm substrates under osteogenic media. In contrast, osteogenic differentiation occurred earlier, and to greater extent, on the 200 nm grain size titania, compared to results obtained on either the 50 or 1500 nm grain sizes, or the glass (reference) surfaces, under osteogenic media. These results demonstrated that biomaterial substrate topography, such as ceramic grain size, affects mesenchymal stem cell differentiation in a size-dependent but, non-linear, manner.     

Apatite–organic polymer composites prepared by a biomimetic process: improvement in adhesion of the apatite layer to the substrate by ultraviolet irradiation

A dense and uniform layer of highly bioactive apatite can be formed in arbitrary thickness on any kind and shape of organic polymer substrates by the following biomimetic process. The substrate is first placed in contact with granular particles of CaO, SiO2-based glass soaked in a simulated body fluid with ion concentrations nearly equal to those of human blood plasma for forming apatite nuclei, and then soaked in another fluid highly supersaturated with respect to the apatite for making the apatite nuclei grow. In the present study, the polymer substrates were pretreated with ultraviolet (UV) light, and then subjected to the biomimetic process described above. By UV irradiation, the induction period for the apatite nucleation of poly(ethylene terephthalate) (PET), poly-ether sulphone (PESF), polyethylene (PE), poly(methyl methacrylate) (PMMA) and polyamide 6 (N6) substrates were reduced form 24 h to 10 h. The adhesive strengths of the apatite layer to the substrates increased from 2.5–3.2 MPa to 4.5–6.0 MPa for PET, PESF and PMMA, and from about 1.0 MPa to 4.0–6.5 MPa for PE and N6 substrates. These results have been explained by assuming that silicate ions, which induce apatite nucleation, are easily adsorbed on the substrates due to the formation of polar groups, with an improved hydrophilic nature, on the polymer surfaces by UV irradiation. © 1998 Chapman & Hall     

Direct evidence of phospholipids in gecko footprints and spatula–substrate contact interface detected using surface-sensitive spectroscopy

Observers ranging from Aristotle to young children have long marvelled at the ability of geckos to cling to walls and ceilings. Detailed studies have revealed that geckos are ‘sticky’ without the use of glue or suction devices. Instead, a gecko''s stickiness derives from van der Waals interactions between proteinaceous hairs called setae and substrate. Here, we present surprising evidence that although geckos do not use glue, a residue is transferred on surfaces as they walk—geckos leave footprints. Using matrix-free nano-assisted laser desorption-ionization mass spectrometry, we identified the residue as phospholipids with phosphocholine head groups. Moreover, interface-sensitive sum-frequency generation spectroscopy revealed predominantly hydrophobic methyl and methylene groups and the complete absence of water at the contact interface between a gecko toe pad and the substrate. The presence of lipids has never been considered in current models of gecko adhesion. Our analysis of gecko footprints and the toe pad–substrate interface has significant consequences for models of gecko adhesion and by extension, the design of synthetic mimics.     

A mathematical model of wound healing and subsequent scarring

Wound healing is a complex process involving the delicate interaction between elements that vary widely in nature and size scales, from the nanometre level, such as molecules, to cells measured in micrometres, and fibres with width and length measured on both scales. Hybrid approaches, where each species is represented by a model on an appropriate size scale, have received attention recently. In this study, we provide a review of earlier work on such hybrid models of wound healing. General models for each of the element types involved in dermal wound healing used in this research are described: cells, modelled as discrete individuals; chemicals, modelled as continua; and fibres, modelled with a novel tensorial representation. Techniques for integrating such disparate models are outlined. A six-species model (fibrin, collagen, macrophages, fibroblasts, transforming growth factor-β (TGF-β) and tissue plasminogen activator) of dermal wound healing is presented. The role of the cytokine TGF-β in the healing cascade is investigated using the model, along with its role in the degree of scarring in the healed tissue.     

The attachment strategy of English ivy: a complex mechanism acting on several hierarchical levels

English ivy (Hedera helix L.) is able to grow on vertical substrates such as trees, rocks and house plaster, thereby attaching so firmly to the surface that when removed by force typically whole pieces of the climbing substrate are torn off. The structural details of the attachment process are not yet entirely understood. We studied the attachment process of English ivy in detail and suggest a four-phase process to describe the attachment strategy: (i) initial physical contact, (ii) form closure of the root with the substrate, (iii) chemical adhesion, and (iv) shape changes of the root hairs and form-closure with the substrate. These four phases and their variations play an important role in the attachment to differently structured surfaces. We demonstrate that, in English ivy, different mechanisms work together to allow the plant''s attachment to various climbing substrates and reveal the importance of micro-fibril orientation in the root hairs for the attachment based on structural changes at the subcellular level.     

Knowing one's place: a free-energy approach to pattern regulation

Understanding how organisms establish their form during embryogenesis and regeneration represents a major knowledge gap in biological pattern formation. It has been recently suggested that morphogenesis could be understood in terms of cellular information processing and the ability of cell groups to model shape. Here, we offer a proof of principle that self-assembly is an emergent property of cells that share a common (genetic and epigenetic) model of organismal form. This behaviour is formulated in terms of variational free-energy minimization—of the sort that has been used to explain action and perception in neuroscience. In brief, casting the minimization of thermodynamic free energy in terms of variational free energy allows one to interpret (the dynamics of) a system as inferring the causes of its inputs—and acting to resolve uncertainty about those causes. This novel perspective on the coordination of migration and differentiation of cells suggests an interpretation of genetic codes as parametrizing a generative model—predicting the signals sensed by cells in the target morphology—and epigenetic processes as the subsequent inversion of that model. This theoretical formulation may complement bottom-up strategies—that currently focus on molecular pathways—with (constructivist) top-down approaches that have proved themselves in neuroscience and cybernetics.     

Nanofibrous membrane of collagen–polycaprolactone for cell growth and tissue regeneration

Nanofibrous substrates of synthetic polymers including polycaprolactone (PCL) have shown considerable potential in tissue regeneration. This paper reports the use of PCL/collagen nanofibers to improve the in vitro osteoblastic responses for the applications in bone regeneration area. Collagen and PCL were dissolved in a co-solvent, and the resulting solution was electrospun into a nanofibrous web. Nonwoven fibrous matrices were successfully produced at various compositional ratios (PCL/collagen = 1/3, 1 and 3 by weight). Although the PCL nanofiber was hydrophobic, the presence of collagen significantly improved the water affinity, such as the water contact angle and water uptake capacity. Tensile mechanical tests showed that the collagen–PCL nanofiber had a significantly higher extension rate (approximately 2.8-fold) than the PCL while maintaining the maximum tensile load in a similar range. The osteoblastic cells cultured on the collagen–PCL nanofibrous substrate showed better initial adhesion and a higher level of growth than those cultured on the PCL nanofiber. Furthermore, real-time RT-PCR revealed the expression of a series of bone-associated genes, including osteopontin, collagen type I and alkaline phosphatase. The expression of these genes was significantly higher on the collagen–PCL nanofiber than on the PCL nanofiber. When subcutaneously implanted in mouse the collagen–PCL membrane facilitated tissue cells to well penetrate into the nanofibrous structure at day 7, whilst no such cell penetration was noticed in the pure PCL nanofiber. Overall, the presence of collagen within the PCL nanofiber improves the water affinity, tensile extension rate, and the tissue cell responses, such as initial adhesion, growth, penetration and the expression of bone-associated genes. Therefore, the collagen–PCL nanofibrous membrane may have potential applications in the cell growth and bone tissue regeneration.