PC Lipids Research Articles

Development of LC-FAIMS-MS and its application to lipidomics study of Acinetobacter baumannii infection

The recent advances in mass spectrometry (MS) technologies have enabled comprehensive lipid profiling in biological samples. However, the robustness and efficiency of MS-based lipidomics is compromised by the complexity of biological samples. High-field asymmetric waveform ion mobility spectrometry (FAIMS) is a technology that can continuously transmit one type of ion, independent of the mass-to-charge ratio. Here we present the development and application of LC-FAIMS-MS/MS-based platform for untargeted lipidomics. We used 3 optimally balanced compensation voltages, i.e., 29 V, 34 V and 39 V, to analyze all subclasses of glycerophospholipids. The reproducibility of the method was evaluated using reference standards. The reproducibility of retention times ranged from 0.9% to 1.5% RSD; whereas RSD values of 5%–10% were observed for peak areas. More importantly, the coupling of a FAIMS device can significantly improve the robustness and efficiency. We exploited this NPLC-FAIMS-HRMS to analyze the serum lipid profiles in mice infected intranasally with Acinetobacter baumannii. The temporal profiles of serum lipids after A. baumannii inoculation were obtained for 4 h, 8 h, and 24 h. We found that nearly all ether PC and ether PE lipids were significantly decreased 8 h after inoculation. The resultant volcano plot illustrated the distribution of 28 increased and 28 decreased lipid species in mouse sera 24 h after inoculation. We also found that a single ether PE composition can comprise multiple isomeric structures, and the relative abundance of each isomer could be quantified using the newly developed NPLC-FAIMS-PRM method.

Candida tropicalis ZD-3 prevents excessive fat deposition by regulating ileal microbiota and bile acids enterohepatic circulation in broilers.

Evidence suggests that the dietary intake of Candida tropicalisZD-3 (ZD-3) has various health benefits, but the treatment mechanisms and effects remain unclear. The aim of this study investigates the effect of ZD-3 on reducing fat deposition in broilers and the underlying mechanism. 180 one-day-old, yellow-feathered broilers were randomly divided into three groups: control (CON) group fed a basal diet, an active Candida tropicalis ZD-3 (ZD) group supplemented with ZD, and a heat-inactivated Candida tropicalis ZD-3 (HZD) group supplemented with HZD. The experiment lasted for 28 d. The ZD and HZD treatments significantly reduced the abdominal fat index (p < 0.05), decreased TG levels in serum and liver (p < 0.05), altered the ileal microbial composition by reducing the Firmicutes to Bacteroidetes (F/B) ratio. Additionally, the ZD and HZD treatments reduced liver cholesterol by decreasing ileal FXR-FGF19 signaling and increasing liver FXR-SHP signaling (p < 0.05). The ZD and HZD treatments also changed liver PC and TG classes lipid composition, regulating liver lipid metabolism by promoting TG degradation and modulating the signal transduction of the cell membrane. Overall, ZD-3 was effective in improving lipid metabolism in broilers by regulating the ileal microbial composition and BAs enterohepatic circulation. This study provides a theoretical basis for the development and application of ZD-3 for the regulation of lipid metabolism in broilers.

Elements of the C-terminal tail of a C-terminal domain homolog of the Orange Carotenoid Protein determining xanthophyll uptake from liposomes

Carotenoids perform multifaceted roles in life ranging from coloration over light harvesting to photoprotection. The Orange Carotenoid Protein (OCP), a light-driven photoswitch involved in cyanobacterial photoprotection, accommodates a ketocarotenoid vital for its function. OCP extracts its ketocarotenoid directly from membranes, or accepts it from homologs of its C-terminal domain (CTDH). The CTDH from Anabaena (AnaCTDH) was shown to be important for carotenoid transfer and delivery from/to membranes. The C-terminal tail of AnaCTDH is a critical structural element likely serving as a gatekeeper and facilitator of carotenoid uptake from membranes. We investigated the impact of amino acid substitutions within the AnaCTDH-CTT on echinenone and canthaxanthin uptake from DOPC and DMPG liposomes. The transfer rate was uniformly reduced for substitutions of Arg-137 and Arg-138 to Gln or Ala, and depended on the lipid type, indicating a weaker interaction particularly with the lipid head group. Our results further suggest that Glu-132 has a membrane-anchoring effect on the PC lipids, specifically at the choline motif as inferred from the strongly different effects of the CTT variants on the extraction from the two liposome types. The substitution of Pro-130 by Gly suggests that the CTT is perpendicular to both the membrane and the main AnaCTDH protein during carotenoid extraction. Finally, the simultaneous mutation of Leu-133, Leu-134 and Leu-136 for alanines showed that the hydrophobicity of the CTT is crucial for carotenoid uptake. Since some substitutions accelerated carotenoid transfer into AnaCTDH while others slowed it down, carotenoprotein properties can be engineered toward the requirements of applications.

Lipid Isobaric Mass Tagging for Enhanced Relative Quantification of Unsaturated sn-Positional Isomers.

Changes in the levels of lipid sn-positional isomers are associated with perturbation of the physiological environment within the biological system. Consequently, knowing the concentrations of these lipids holds significant importance for unraveling their involvement in disease diagnosis and pathological mechanisms. However, existing methods for lipid quantification often fall short in accuracy due to the structural diversity and isomeric forms of lipids. To address this challenge, we have developed an aziridine-based isobaric tag labeling strategy that allows (i) differentiation and (ii) enhanced relative quantification of lipid sn-positional isomers from distinct samples in a single run. The methodology enabled by aziridination, isobaric tag labeling, and lithiation has been applied to various phospholipids, enabling the determination of the sn-positions of fatty acyl chains and enhanced relative quantification. The analysis of Escherichia coli lipid extracts demonstrated the enhanced determination of the concentration ratios of lipid isomers by measuring the intensity ratios of mass reporters released from sn-positional diagnostic ions. Moreover, we applied the method to the analysis of human colon cancer plasma. Intriguingly, 17 PC lipid sn-positional isomers were identified and quantified simultaneously, and among them, 7 showed significant abundance changes in the colon cancer plasma, which can be used as potential plasma markers for diagnosis of human colon cancer.

Digestion of lipid micelles leads to increased membrane permeability.

Lipid-based drug carriers are an attractive option to solubilise poorly water soluble therapeutics. Previously, we reported that the digestion of a short tail PC lipid (2C6PC) by the PLA2 enzyme has a significant effect on the structure and stability of the micelles it forms. Here, we studied the interactions of micelles of varying composition representing various degrees of digestion with a model ordered (70 mol% DPPC & 30 mol% cholesterol) and disordered (100% DOPC) lipid membrane. Micelles of all compositions disassociated when interacting with the two different membranes. As the percentage of digestion products (C6FA and C6LYSO) in the micelle increased, the disassociation occurred more rapidly. The C6FA inserts preferentially into both membranes. We find that all micelle components increase the area per lipid, increase the disorder and decrease the thickness of the membranes, and the 2C6PC lipid molecules have the most significant impact. Additionally, there is an increase in permeation of water into the membrane that accompanies the insertion of C6FA into the DOPC membranes. We show that the natural digestion of lipid micelles result in molecular species that can enhance the permeability of lipid membranes that in turn result in an enhanced delivery of drugs.

Lipid bilayer properties potentially contributed to the evolutionary disappearance of betaine lipids in seed plants

BackgroundMany organisms rely on mineral nutrients taken directly from the soil or aquatic environment, and therefore, developed mechanisms to cope with the limitation of a given essential nutrient. For example, photosynthetic cells have well-defined responses to phosphate limitation, including the replacement of cellular membrane phospholipids with non-phosphorous lipids. Under phosphate starvation, phospholipids in extraplastidial membranes are replaced by betaine lipids in microalgae. In higher plants, the synthesis of betaine lipid is lost, driving plants to other strategies to cope with phosphate starvation where they replace their phospholipids by glycolipids.ResultsThe aim of this work was to evaluate to what extent betaine lipids and PC lipids share physicochemical properties and could substitute for each other. By neutron diffraction experiments and dynamic molecular simulation of two synthetic lipids, the dipalmitoylphosphatidylcholine (DPPC) and the dipalmitoyl-diacylglyceryl-N,N,N-trimethylhomoserine (DP-DGTS), we found that DP-DGTS bilayers are thicker than DPPC bilayers and therefore are more rigid. Furthermore, DP-DGTS bilayers are more repulsive, especially at long range, maybe due to unexpected unscreened electrostatic contribution. Finally, DP-DGTS bilayers could coexist in the gel and fluid phases.ConclusionThe different properties and hydration responses of PC and DGTS provide an explanation for the diversity of betaine lipids observed in marine organisms and for their disappearance in seed plants.

Metabolomic, lipidomic, and proteomic profiles provide insights on meat quality differences between Shitou and Wuzong geese

A comprehensive comparison of metabolomic, lipidomic, and proteomic profiles was conducted between the breast and leg muscles of Shitou goose (STE) and Wuzhong goose (WZE), which exhibit significant variations in body size and growth rate, to evaluate their impact on meat quality. WZE had higher intramuscular fat content in their breast muscles, which were also chewier and had higher drip and cooking losses than STE. Metabolomic analysis revealed differential regulation of amino acid and purine metabolism between WZE and STE. Lipidomic analysis indicated a higher abundance of PE and PC lipid molecules in WZE. Integration of proteomic and metabolomic data highlighted purine metabolism and amino acid biosynthesis as the major distinguishing pathways between STE and WZE. The primary differential pathways between breast and leg muscles were associated with energy metabolism and fatty acid metabolism. This comprehensive analysis provides valuable insights into the distinct meat quality of STE and WZE.

Lipid composition and properties affect protein-mediated carotenoid uptake efficiency from membranes

Carotenoids are pigments of diverse functions ranging from coloration over light-harvesting to photoprotection. Yet, the number of carotenoid-binding proteins, which mobilize these pigments in physiological media, is limited, and the mechanisms of carotenoid mobilization are still not well understood. The same applies for the determinants of carotenoid uptake from membranes into carotenoproteins, especially regarding the dependence on the chemical properties of membrane lipids. Here, we investigate xanthophyll uptake capacity and kinetics of a paradigmatic carotenoid-binding protein, the homolog of the Orange Carotenoid Protein's C-terminal domain from Anabaena sp. PCC 7120 (AnaCTDH), using liposomes formed from defined lipid species and loaded with canthaxanthin (CAN) and echinenone (ECN), respectively. Phospholipids with different chain length and degree of saturation were investigated. The composition of carotenoid-loaded liposomes directly affected the incorporation yield and storage ratio of CAN and ECN as well as the rate of carotenoid uptake by AnaCTDH. Generally, saturated PC lipids were identified as unsuitable, and a high phase transition temperature of the lipids negatively affected the carotenoid incorporation and storage yield. For efficient carotenoid transfer, the velocity increases with increasing chain length or membrane thickness. An average transfer yield of 93 % and 43 % were obtained for the formation of AnaCTDH(CAN) and AnaCTDH(ECN) holoproteins, respectively. In summary, the most suitable lipids for the formation of AnaCTDH(CAN/ECN) holoproteins by carotenoid transfer from artificial liposomes are phosphatidylcholine (18:1) and phosphatidylglycerol (14:0). Thus, these two lipids provide the best conditions for further investigation of lipid-protein interaction and the carotenoid uptake process.

Dual metal electrolysis in theta capillary for lipid analysis

Increasing studies associating glycerophospholipids with various pathological conditions highlight the need for their thorough characterization. However, the intricate composition of the lipidome due to the presence of lipid isomers poses significant challenges to structural lipidomics. This study uses the anodic corrosion of two metals in a single theta nESI emitter as a tool to simultaneously characterize lipids at multiple isomer levels. Anodic corrosion of cobalt and copper in the positive ion mode generates the metal-adducted lipid complexes, [M+Co]2+ and [M+Cu]+, respectively. Optimization of parameters such as the distances of the electrodes from the nESI tip allowed the achievement of the formation of one metal-adducted lipid product at a time. Collision-induced dissociation (CID) of [M+Co]2+ results in preferential loss of the fatty acyl (FA) chain at the sn-2 position, thus generating singly charged sn-specific fragment ions. Whereas, multistage fragmentation of [M+Cu]+ via CID generated a CC bond position-specific characteristic ion pattern induced by the π-Cu+ interaction. The feasibility of the method was tested on PC lipid extract from egg yolk to identify lipids on multiple isomer levels. Thus, the application of dual metal anodic corrosion allows lipid isomer identification with reduced sample preparation time, no signal suppression by counter anions, low sample consumption, and no need for an extra apparatus.

Location of dopamine in lipid bilayers and its relevance to neuromodulator function

Dopamine (DA) is a neurotransmitter that also acts as a neuromodulator, with both functions being essential to brain function. Here, we present the first experimental measurement of DA location in lipid bilayers using x-ray diffuse scattering, solid-state deuterium NMR, and electron paramagnetic resonance. We find that the association of DA with lipid headgroups as seen in electron density profiles leads to an increase of intermembrane repulsion most likely due to electrostatic charging. DA location in the lipid headgroup region also leads to an increase of the cross-sectional area per lipid without affecting the bending rigidity significantly. The order parameters measured by solid-state deuterium NMR decrease in the presence of DA for the acyl chains of PC and PS lipids, consistent with an increase in the area per lipid due to DA. Most importantly, these results support the hypothesis that three-dimensional diffusion of DA to target membranes could be followed by relatively more efficient two-dimensional diffusion to receptors within those membranes.

Nanoscale membrane dynamics in chain asymmetric lipid bilayers.

Lipids that have two tails of different lengths are found throughout biomembranes in nature, yet the effects of this asymmetry on the membrane properties are not well understood, especially the membrane dynamics. Accommodating such a difference in tail length could significantly impact the membrane dynamics and elastic properties depending on how the longer tail is situated within the bilayer. To probe these effects, here we study the nanoscale bending fluctuations in model chain-asymmetric 14:0-18:0 PC (MSPC) and 18:0-14:0 PC (SMPC) lipid bilayers using neutron spin echo (NSE) spectroscopy. We find that despite the partial interdigitation that is known to persist in the fluid phase of these membranes, the collective fluctuations are enhanced on timescales of tens of nanoseconds, and the chain-asymmetric lipid bilayers are softer than an analogous chain-symmetric lipid with the same average number of carbons in the acyl tails, di-16:0 PC (DPPC). Quantitative comparison of the NSE results suggests that the enhanced bending fluctuations at the nanosecond timescales are consistent with experimental and computational studies that showed the compressibility moduli of chain-asymmetric lipid membranes are 20 % to 40% lower than chain-symmetric lipid membranes. These studies add to growing evidence that the partial interdigitation in chain-asymmetric lipid membranes in the fluid phase is highly dynamic and influences membrane dynamic processes from the molecular to mesoscopic length scales without significantly changing the bilayer thickness or area per lipid.

Effects of interdigitation on the biophysical properties of lipid bilayers: A molecular dynamics study of chain asymmetric lipids.

Plasma membranes (PMs) are composed of hundreds of different lipids varying in structure and chemical properties. A major challenge in membrane biophysics is to determine how different classes of lipids contribute to the physical properties of the PM. One particularly interesting type of lipid is characterized by a different number of carbons in its two hydrocarbon chains. These chain asymmetric lipids are thought to have a higher propensity for interdigitation with lipids in the opposing leaflet and thus, to have a unique effect on interleaflet coupling. We have used molecular dynamics (MD) simulations to systematically examine fluid-phase bilayers composed of fully saturated PC lipids, each with a total of 32 acyl chain carbons, but increasing asymmetry ranging from zero to four carbons difference between the two chains. We quantified interdigitation in these bilayers using three novel methods and compared the results to standard approaches; we found general agreement in the trends, namely that increasing chain asymmetry results in increased interdigitation. We hypothesized that a greater extent of interdigitation should result in stiffer bilayers and thus, higher values of the bending modulus. To test this hypothesis, we measured various structural and mechanical properties of the simulated bilayers including thickness, lipid packing, and bending and compressibility moduli. Surprisingly, we found that the bilayers with the greatest extent of interdigitation are also the softest at a common temperature of 50°C. These results provide new insights into the structural and mechanical effects of lipid chain asymmetry and open up further questions about the nature of the coupling between lipid chain interdigitation and membrane biophysical properties.

Nanoscale Bending Dynamics in Mixed-Chain Lipid Membranes

Lipids that have two tails of different lengths are found throughout biomembranes in nature, yet the effects of this asymmetry on the membrane properties are not well understood, especially when it comes to the membrane dynamics. Here we study the nanoscale bending fluctuations in model mixed-chain 14:0–18:0 PC (MSPC) and 18:0–14:0 PC (SMPC) lipid bilayers using neutron spin echo (NSE) spectroscopy. We find that despite the partial interdigitation that is known to persist in the fluid phase of these membranes, the collective fluctuations are enhanced on timescales of tens of nanoseconds, and the chain-asymmetric lipid bilayers are softer than an analogous chain-symmetric lipid bilayer with the same average number of carbons in the acyl tails, di-16:0 PC (DPPC). Quantitative comparison of the NSE results suggests that the enhanced bending fluctuations at the nanosecond timescales are consistent with experimental and computational studies that showed the compressibility moduli of chain-asymmetric lipid membranes are 20% to 40% lower than chain-symmetric lipid membranes. These studies add to growing evidence that the partial interdigitation in mixed-chain lipid membranes is highly dynamic in the fluid phase and impacts membrane dynamic processes from the molecular to mesoscopic length scales without significantly changing the bilayer thickness or area per lipid.

Structure of the gel phase of diC22:1PC lipid bilayers determined by x-ray diffraction

High-resolution x-ray data are reported for the ordered phases of long-chain di-monounsaturated C22:1 phosphocholine lipid bilayers. Similar to PC lipids that have saturated chains, diC22:1PC has a subgel phase and a gel phase, but dissimilarly, we find no ripple phase. Our quantitative focus is on the structure of the gel phase. We have recorded 17 lamellar orders, indicating a very well-ordered structure. Fitting to a model provides the phases of the orders. The Fourier construction of the electron density profile has two well-defined headgroup peaks and a very sharp and deep methyl trough. The wide-angle scattering exhibits two Bragg rods that provide the area per molecule. They have an intensity pattern quite different than that of lipids with saturated chains. Models of chain packing indicate that ground state chain configurations are tilted primarily toward next nearest neighbors with an angle that is also consistent with the modeling of the electron density profile. Wide-angle modeling also indicates broken mirror symmetry between the monolayers. Our wide-angle results and our electron density profile together leads to the hypothesis that the sn-1 and sn-2 chains have equivalent penetration depths in contrast to the gel phase structure of lipids with saturated hydrocarbon chains.

Preparation of Giant Quantum Dot-Liposome Complexes by the Asolectin Lipid and Theoretical Model for Stabilization of Nanoparticle Inside the Liposome

We prepare giant Quantum dot-Liposome Complexes (QLCs). Quantum dots (QDs) incorporated inside liposome above 10 μm. QLCs is made by using the electro-swelling method combined with spin coating techniques. Three types of PC lipids and asolectin lipid are used for QLCs with HDA (hexadecylamine) coated QDs, which ranged from blue- (diameter ~2.1 nm) to red-emission (diameter ~5.0 nm). As expected, (blue- or) green-emission QDs (smaller than) comparable to the thickness of PC lipid bilayer (~4 nm) are successfully formed QLCs, but QDs bigger than that fail to reproduce. This observation is well-consistent with those reported by Gopakumar et al. Surprisingly, all QDs irrespective of their size are, contrary to PC lipids, successfully loaded into asolectin lipid bilayer. In order to understand what makes different behaviors between PC and asolectin lipids on QLC formation, we suggest a theoretical model based on a geometrical assumptions for deformed lipid bilayer surrounding QD. The main advantage of this model is that the critical size Rcr of QD radius can be decided without calculating elastic free energy. As a result, it predicts that only QDs below the critical size (diameter ~3.0 nm) can be loaded in a typical PC-lipid, but all size of QDs can be incorporated into asolectin bilayer under the assumption of two different curvatures on deformed monolayer.

Dilute Bicelles for Glycosyltransferase Studies, Novel Bicelles with Phosphatidylinositol.

Solution-state NMR can be used to study protein–lipid interactions, in particular, the effect that proteins have on lipids. One drawback is that only small assemblies can be studied, and therefore, fast-tumbling bicelles are commonly used. Bicelles contain a lipid bilayer that is solubilized by detergents. A complication is that they are only stable at high concentrations, exceeding the CMC of the detergent. This issue has previously been addressed by introducing a detergent (Cyclosfos-6) with a substantially lower CMC. Here, we developed a set of bicelles using this detergent for studies of membrane-associated mycobacterial proteins, for example, PimA, a key enzyme for bacterial growth. To mimic the lipid composition of mycobacterial membranes, PI, PG, and PC lipids were used. Diffusion NMR was used to characterize the bicelles, and spin relaxation was used to measure the dynamic properties of the lipids. The results suggest that bicelles are formed, although they are smaller than “conventional” bicelles. Moreover, we studied the effect of MTSL-labeled PimA on bicelles containing PI and PC. The paramagnetic label was shown to have a shallow location in the bicelle, affecting the glycerol backbone of the lipids. We foresee that these bicelles will be useful for detailed studies of protein–lipid interactions.

Structure of gel phase di-C22:1PC lipid bilayers

There has been a dearth of structural studies of the low temperature bilayer phases of unsaturated phophocholine lipids, partly because most of them have inconveniently low main phase transition temperatures TM as temperature is lowered from the fluid phase. This paper studies the phase behavior of bilayers of the long chain diC22:1PC lipids and the structure at temperatures below the TM of 13 °C. We were curious whether a ripple phase forms below TM, and whether there are gel and subgel phases like those that occur in saturated chain PC lipid bilayers.

Molecular dynamics simulation study of the positioning and dynamics of α-tocopherol in phospholipid bilayers.

Using molecular dynamics simulations, we investigate the interaction of α-tocopherol (α-toc) with dipalmitoylphosphatidylcholine (DPPC), dimyristoylphosphatidylcholine (DMPC), palmitoyloleoylphosphatidylcholine (POPC), and palmitoyloleoylphosphatidylethanolamine (POPE) lipid bilayers. The goal is to develop a better understanding of the positioning and orientation of α-toc inside the bilayers; properties of significant relevance to α-toc anti-oxidant activity. We investigated bilayer systems with 128 lipids in the presence of either single or 14 α-toc molecules. The single α-toc bilayer systems were investigated via biased MD simulations in which the potential of mean force (PMF) and diffusivity were obtained as functions of the distance between α-toc head group and bilayer center. The higher α-toc concentration systems were investigated with unbiased MD simulations. For all four bilayers at both concentrations, the simulations show that the most probable location of the α-toc hydroxyl group is just below the lipid carbonyl group. Overall, the simulation results are in good agreement with existing experimental data except for the DMPC bilayer system for which some experiments predict α-toc to be located closer to bilayer center. The flip-flop frequency calculated shows that the α-toc flip-flop rate is sensitive to bilayer lipid type. In particular, α-toc has a much lower flip-flop rate in a POPE bilayer compared to the three PC lipid bilayers due to the smaller area per lipid in the POPE bilayer. For DMPC and POPC, the α-toc flip-flop rates are significantly higher at higher α-toc concentration and this appears to be related to the local structural disruption caused by α-toc clusters spanning the bilayer.

Specificity of Loxosceles α Clade Phospholipase D Enzymes for Choline-Containing Lipids: Role of a Conserved Aromatic Cage

Phospholipase D (PLD) enzymes are one of the major toxins in the recluse spider (genus Loxosceles) venom. Loxosceles PLD enzymes are classified in either the α clade or the β clade, and some correlation exists between α or β clade membership and high or low catalytic activity against sphingomyelin (SM), respectively. Lajoie et al., recently showed that a β clade enzyme from Sicarius terrosus (St_βID1) had a strong preference for substrates containing ethanolamine headgroups, and suggested that this substrate specificity originated from the interactions between the PLD interfacial face and lipids. To further understand the substrate specificity of these enzymes, we performed simulations of two PLDs from the α clade, Loxosceles intermedia αIA1 (Li_αIA1) and Loxosceles laeta αIII1 (Ll_αIII1), and one from the β clade, St_βIB1i. Simulations were performed in the presence of two types of lipid bilayers, choline-containing and ethanolamine-containing bilayer. We observed that the two α clade PLDs bound to bilayers with choline-containing lipids (PC or SM) using the catalytic loop. By contrast, St_ βIB1i, did not bind to those bilayers. Analyses of the trajectories also reveal the importance, for the two α clade PLDs, of three aromatic residues located on the catalytic loop and forming a π-cage interacting with a choline group. A multiple sequence alignment of 18 PLDs of both clades reveal that the aromatic cage is conserved in α clade PLDs and absent from at least one major subgroup of the β clade enzymes. We suggest that this cage controls the affinity for choline headgroups, which is in agreement with earlier reports of PC lipid headgroups interacting with aromatic amino acids.

Thermal Reversal Surface with “Sticky Tentacle” for Modulating Initial Cell Adhesion and Detachment

Thermal responsive polymers were introduced for effectively cell detachment from the extracellular matrix (ECM), where the poor cell adhesive capacity was improved by the immobilized biodhesion molecules (most are proteins) at the termini of grafted polymer. However, this protein-mediated adhesion makes it hard to be precisely controlled and takes longer time to start initial cell attachment. Here we describe a choline phosphate(CP) modified thermal responsive surface with enhanced initial cell adhesion based on multivalent CP-PC (phosphatidyl choline, the head-group of phospholipids in all the eukaryotic cell membranes) interaction. The well-defined functional surfaces were constructed using surface-initiated ATRP and click reaction, where the initial cell attachment reach saturation in 10 min and can be regulated by CP density on the surface. Consequently, the fast cell adhesion significantly accelerates cell spreading and proliferation. The facile cell harvest, as opposed to the original mechanism of thermally-induced cell detachment, is to take advantage of the thermal contraction of the polymer above the LCST, which release the adherent cell by reducing the accessibility of the CP groups to cell membrane PC lipids. Si-CP is expected to provide a promising universal structural model for modulating cell attachment and detachment in tissue engineering and regenerative medicine field.