Area Expansion Modulus Research Articles

Thicket and Mesh: How the Outer Membrane Can Resist Tension Imposed by the Cell Wall.

The cell envelope of Gram-negative bacteria is composed of an outer membrane (OM) and an inner membrane (IM) and a peptidoglycan cell wall (CW) between them. Combined with Braun's lipoprotein (Lpp), which connects the OM and the CW, and numerous membrane proteins that exist in both OM and IM, the cell envelope creates a mechanically stable environment that resists various physical and chemical perturbations to the cell, including turgor pressure caused by the solute concentration difference between the cytoplasm of the cell and the extracellular environment. Previous computational studies have explored how individual components (OM, IM, and CW) can resist turgor pressure although combinations of them have been less well studied. To that end, we constructed multiple OM-CW systems, including the Lpp connections with the CW under increasing degrees of strain. The results show that the OM can effectively resist the tension imposed by the CW, shrinking by only 3-5% in area even when the CW is stretched to 2.5× its relaxed area. The area expansion modulus of the system increases with increasing CW strain, although the OM remains a significant contributor to the envelope's mechanical stability. Additionally, we find that when the protein TolC is embedded in the OM, its stiffness increases.

Polyunsaturated fatty acids differentially modulate membrane elasticity in membranes containing high cholesterol

Membrane composition is important for the function of membrane proteins including ion channels and membrane-embedded receptors. One class of proteins that are especially sensitive to membrane composition are mechanosensitive channels, which activate in response to changes in membrane tension. Recently, polyunsaturated fatty acids (PUFAs) were demonstrated to alter mechanosensitive channel behavior, but the mechanism for these observations remains unknown. Mechanosensitive channel behavior is expected to be dependent on membrane mechanical properties such as the membrane area expansion modulus (Ka), which measures the extent to which membrane tension exerts a change in bilayer area.

EPA and DHA differentially modulate membrane elasticity in the presence of cholesterol

Polyunsaturated fatty acids (PUFAs) modify the activity of a wide range of membrane proteins and are increasingly hypothesized to modulate protein activity by indirectly altering membrane physical properties. Among the various physical properties affected by PUFAs, the membrane area expansion modulus (Ka), which measures membrane strain in response to applied force, is expected to be a significant controller of channel activity. Yet, the impact of PUFAs on membrane Ka has not been measured previously. Through a series of micropipette aspiration studies, we measured the apparent Ka (Kapp) of phospholipid model membranes containing nonesterified fatty acids. First, we measured membrane Kapp as a function of the location of the unsaturated bonds and degree of unsaturation in the incorporated fatty acids and found that Kapp generally decreases in the presence of fatty acids with three or more unsaturated bonds. Next, we assessed how select ω-3 PUFAs, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), affect the Kapp of membranes containing cholesterol. In vesicles prepared with high amounts of cholesterol, which should increase the propensity of the membrane to phase segregate, we found that inclusion of DHA decreases the Kapp in comparison to EPA. We also measured how these ω-3 PUFAs affect membrane fluidity and bending rigidity to determine how membrane Kapp changes in relation to these other physical properties. Our study shows that PUFAs generally decrease the Kapp of membranes and that EPA and DHA have differential effects on Kapp when membranes contain higher levels of cholesterol. Our results suggest membrane phase behavior and the distribution of membrane-elasticizing amphiphiles impact the ability of a membrane to stretch.

Fat inclusions strongly alter membrane mechanics

Neutral lipids (NLs) are apolar oil molecules synthesized in the endoplasmic reticulum bilayer upon diverse biological stimuli. NLs synthesized are released in the hydrophobic core of the bilayer. At a critical concentration, NLs condense by phase separation and nucleate a lipid droplet (LD). After an LD forms, a fraction of NLs can be present in the bilayer but at a concentration below that of the nucleation. Here, we study whether and how the accumulation of NLs alters a lipid bilayer’s mechanical properties. In synthetic systems, we found that NLs proffer unusual bilayer stretching capacities, especially in the presence of negatively curved phospholipids. This impact becomes spectacular when an LD is contiguous with the bilayer and supplies it with NLs. The tested NLs markedly decrease the bilayer area expansion modulus and significantly increase lysis tension but had opposite effects on membrane bending rigidity. Our data unveil how NL molecules modify overall membrane mechanics, the alteration of which may be linked to pathologies or anticancer treatments targeting NLs.

Quantifying the Effect of Fatty Acids on the Elasticity of Model Membranes

Dietary fatty acids function in many processes that impact cardiovascular health. Specifically, polyunsaturated fatty acids (PUFAs), are expected to improve cardiovascular function through the alteration of membrane properties of diverse cell types. Global membrane properties have increasingly been shown to impact the activity of distinct mechanosensitive membrane proteins. However, the mechanism by which these properties affect mechanosensitive channel activity remains unclear. PUFAs are hypothesized to alter bilayer mechanical properties but monoacyl fatty acids have been studied sparingly in phospholipid bilayers. Moreover, the effect of monoacyl fatty acid chain length, unsaturation, and location of unsaturation on membrane mechanical properties have not yet been determined. With these questions in mind, we incorporated monoacyl PUFAs of different chain lengths and degrees of unsaturation into phospholipid bilayers. Using micropipette aspiration and fluorescence anisotropy, we measured the effect of various PUFAs on the membrane area expansion modulus and fluidity, respectively, of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) vesicle membranes. By systematically altering the identity of fatty acids, we observed that long-chain omega-3 fatty acids—but not monounsaturated fatty acids—reduce the membrane area expansion modulus and increase the fluidity of DOPC vesicle membranes. In addition, we observe that two major omega-3 fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), impact the area expansion modulus and fluidity of DOPC membranes containing cholesterol domains in different ways. These results provide insight into how EPA and DHA may have distinct effects on the bilayer properties of mammalian cells, despite being chemically similar. Our results demonstrate several ways in which dietary monoacyl fatty acids impact the mechanical properties of phospholipid bilayers and present a potential route through which fatty acid uptake into cell membranes may influence the activity of membrane proteins.

Membrane stiffness is one of the key determinants of E. coli MscS channel mechanosensitivity

Mechanosensitive (MS) channels have an intimate relationship with membrane lipids that underlie their mechanosensitivity. Membrane lipids may influence channel activity by directly interacting with MS channels or by influencing the global properties of the membrane such as elastic area expansion modulus or bending rigidity. Previous work has implicated membrane stiffness as a potential determinant of the mechanosensitivity of E. coli (Ec)MscS. Here we systematically tested this hypothesis using patch fluorometry of azolectin liposomes doped with lipids of increasing elastic area expansion modulus. Increasing dioleoylphosphatidylethanolamine (DOPE) content of azolectin liposomes made it more difficult to activate EcMscS by membrane tension (i.e. increased gating threshold). This effect was exacerbated by stiffer forms of phosphatidylethanolamine such as the branched chain lipid diphytanoylphosphoethanolamine (DPhPE) or the fully saturated lipid distearoyl-sn-glycero-3-phosphoethanolamine (DSPE). Furthermore, a comparison of the branched chain lipid diphytanoylphosphocholine (DPhPC) to the stiffer DPhPE indicated again that it was harder to activate EcMscS in the presence of the stiffer DPhPE. We show that these effects are not due to changes in membrane bending rigidity as the membrane tension threshold of EcMscS in membranes doped with PC18:1 and PC18:3 remained the same, despite a two-fold difference in their bending rigidity. We also show that after prolonged pressure application sudden removal of force in softer membranes caused a rebound reactivation of EcMscS and we discuss the relevance of this phenomenon to bacterial osmoregulation. Collectively, our data suggests that membrane stiffness (elastic area expansion modulus) is one of the key determinants of the mechanosensitivity of EcMscS.

Differential or Curvature Stress? Modus Vivendi

Differential or Curvature Stress? Modus Vivendi

Diblock copolymers enhance folding of a mechanosensitive membrane protein during cell-free expression

The expression and integration of membrane proteins into vesicle membranes is a critical step in the design of cell-mimetic biosensors, bioreactors, and artificial cells. While membrane proteins have been integrated into a variety of nonnatural membranes, the effects of the chemical and physical properties of these vesicle membranes on protein behavior remain largely unknown. Nonnatural amphiphiles, such as diblock copolymers, provide an interface that can be synthetically controlled to better investigate this relationship. Here, we focus on the initial step in a membrane protein's life cycle: expression and folding. We observe improvements in both the folding and overall production of a model mechanosensitive channel protein, the mechanosensitive channel of large conductance, during cell-free reactions when vesicles containing diblock copolymers are present. By systematically tuning the membrane composition of vesicles through incorporation of a poly(ethylene oxide)-b-poly(butadiene) diblock copolymer, we show that membrane protein folding and production can be improved over that observed in traditional lipid vesicles. We then reproduce this effect with an alternate membrane-elasticizing molecule, C12E8 Our results suggest that global membrane physical properties, specifically available membrane surface area and the membrane area expansion modulus, significantly influence the folding and yield of a membrane protein. Furthermore, our results set the stage for explorations into how nonnatural membrane amphiphiles can be used to both study and enhance the production of biological membrane proteins.

Dewetting-induced formation and mechanical properties of synthetic bacterial outer membrane models (GUVs) with controlled inner-leaflet lipid composition.

The double-membrane cellular envelope of Gram-negative bacteria enables them to endure harsh environments and represents a barrier to many clinically available antibiotics. The outer membrane (OM) is exposed to the environment and is the first point of contact involved in bacterial processes such as signaling, pathogenesis, and motility. As in the cytoplasmic membrane, the OM in Gram-negative bacteria has a phospholipid-rich inner leaflet and an outer leaflet that is predominantly composed of lipopolysaccharide (LPS). We report on a microfluidic technique for fabricating monodisperse asymmetric giant unilamellar vesicles (GUVs) possessing the Gram-negative bacterial OM lipid composition. Our continuous microfluidic fabrication technique generates 50-150 μm diameter water-in-oil-in-water double emulsions at high-throughput. The water-oil and oil-water interfaces facilitate the self-assembly of phospholipid and LPS molecules to create the inner and outer leaflets of the lipid bilayer, respectively. The double emulsions have ultrathin oil shells, which minimizes the amount of residual organic solvent that remains trapped between the leaflets of the GUV membrane. An extraction process by ethanol and micropipette aspiration of the ultrathin oil shells triggers an adhesive interaction between the two lipid monolayers assembled on the water-oil and oil-water interfaces (i.e., dewetting transition), forcing them to contact and form a lipid bilayer membrane. The effect of different inner-leaflet lipid compositions on the emulsion/vesicle stability and the dewetting transition is investigated. We also report on the values for bending and area expansion moduli of synthetic asymmetric model membranes with lipid composition/architecture that is physiologically relevant to the OM in Pseudomonas aeruginosa bacteria.

Evolutionary specialization of MscCG, an MscS-like mechanosensitive channel, in amino acid transport in Corynebacterium glutamicum

MscCG, a mechanosensitive channel of Corynebacterium glutamicum provides a major export mechanism for glutamate in this Gram-positive bacterium, which has for many years been used for industrial production of glutamate and other amino acids. The functional characterization of MscCG is therefore, of great significance to understand its conductive properties for different amino acids. Here we report the first successful giant spheroplast preparation of C. glutamicum amenable to the patch clamp technique, which enabled us to investigate mechanosensitive channel activities of MscCG in the native membrane of this bacterium. Single channel recordings from these spheroplasts revealed the presence of three types of mechanosensitive channels, MscCG, MscCG2, and CgMscL, which differ largely from each other in their conductance and mechanosensitivity. MscCG has a relatively small conductance of ~340 pS followed by an intermediate MscCG2 conductance of ~1.0 nS and comparably very large conductance of 3.7 nS exhibited by CgMscL. By applying Laplace’s law, we determined that very moderate membrane tension of ~5.5 mN/m was required for half activation of MscCG compared to ~12 mN/m required for half activation of both MscCG2 and CgMscL. Furthermore, by combining the micropipette aspiration technique with molecular dynamics simulations we measured mechanical properties of the C. glutamicum membrane, whose area elasticity module of KA ≈ 15 mN/m is characteristic of a very soft membrane compared to the three times larger area expansion modulus of KA ≈ 44 mN/m of the more elastic E. coli membrane. Moreover, we demonstrate that the “soft” properties of the C. glutamicum membrane have a significant impact on the MscCG gating characterized by a strong voltage-dependent hysteresis in the membrane of C. glutamicum compared to a complete absence of the hysteresis in the E. coli cell membrane. We thus propose that MscCG has evolved and adapted as an MscS-like channel to the mechanical properties of the C. glutamicum membrane enabling the channel to specialize in transport of amino acids such as glutamate, which are major osmolytes helping the bacterial cells survive extreme osmotic stress.

The Interplay of Structure and Dynamics in Mixed Lipid Bilayers

The implication of lipids in membrane mediated processes has motivated significant theoretical and experimental research interest to understand the functional significance of lipid chemical structure variations on the biomembrane structure and dynamics. In particular, hydrophobic mismatch between lipids with different acyl chain lengths is expected to play an important role in determining the local membrane structure and collective membrane dynamics. Herein we investigated these phenomena in large unilamellar vesicles composed of binary mixtures of lipids with a 4 carbon mismatch in tail length, dimyristoylphosphatidylcholine (DMPC, 14:0 C) and distearoylphosphatidycholine (DSPC, 18:0 C), using neutron scattering techniques. Structural studies using small angle neutron scattering (SANS) revealed that the mixed lipid bilayers were thinner than expected for a simple composition-weighted average of the pure component bilayers. The structural differences were accompanied by dramatic differences in the mixed lipid membrane dynamics. Neutron spin echo spectroscopy (NSE) experiments showed that the mixed lipid bilayers were more dynamic than their single component analogs, with the mixed bilayers having a lower bending modulus and increased thickness fluctuation amplitude than the pure DMPC or DSPC bilayers. Both of these enhanced dynamics were consistent with a decrease in the area expansion modulus of the mixed systems. Together our results demonstrate the influence of lipid composition on the bilayer biophysical properties and highlight the complex interplay between structure and dynamics in lipid membranes.

Mechanical Characterization of Hybrid Vesicles Based on Linear Poly(Dimethylsiloxane-b-Ethylene Oxide) and Poly(Butadiene-b-Ethylene Oxide) Block Copolymers.

Poly(dimethylsiloxane-ethylene oxide) (PDMS-PEO) and poly(butadiene-b-ethylene oxide) (PBd-PEO) are two block copolymers which separately form vesicles with disparate membrane permeabilities and fluidities. Thus, hybrid vesicles formed from both PDMS-PEO and PBd-PEO may ultimately allow for systematic, application-specific tuning of vesicle membrane fluidity and permeability. However, given the relatively low strength previously noted for comb-type PDMS-PEO vesicles, the mechanical robustness of the resulting hybrid vesicles must first be confirmed. Toward this end, we have characterized the mechanical behavior of vesicles formed from mixtures of linear PDMS-PEO and linear PBd-PEO using micropipette aspiration. Tension versus strain plots of pure PDMS12-PEO46 vesicles revealed a non-linear response in the high tension regime, in contrast to the approximately linear response of pure PBd33-PEO20 vesicles. Remarkably, the area expansion modulus, critical tension, and cohesive energy density of PDMS12-PEO46 vesicles were each significantly greater than for PBd33-PEO20 vesicles, although critical strain was not significantly different between these vesicle types. PDMS12-PEO46/PBd33-PEO20 hybrid vesicles generally displayed graded responses in between that of the pure component vesicles. Thus, the PDMS12-PEO46/PBd33-PEO20 hybrid vesicles retained or exceeded the strength and toughness characteristic of pure PBd-PEO vesicles, indicating that future assessment of the membrane permeability and fluidity of these hybrid vesicles may be warranted.

Membrane mechanical properties of synthetic asymmetric phospholipid vesicles.

Synthetic lipid vesicles have served as important model systems to study cellular membrane biology. Research has shown that the mechanical properties of bilayer membranes significantly affects their biological behavior. The properties of a lipid bilayer are governed by lipid acyl chain length, headgroup type, and the presence of membrane proteins. However, few studies have explored how membrane architecture, in particular trans-bilayer lipid asymmetry, influences membrane mechanical properties. In this study, we investigated the effects of lipid bilayer architecture (i.e. asymmetry) on the mechanical properties of biological membranes. This was achieved using a customized micropipette aspiration system and a novel microfluidic technique previously developed by our team for building asymmetric phospholipid vesicles with tailored bilayer architecture. We found that the bending modulus and area expansion modulus of the synthetic asymmetric bilayers were up to 50% larger than the values acquired for symmetric bilayers. This was caused by the dissimilar lipid distribution in each leaflet of the bilayer for the asymmetric membrane. To the best of our knowledge, this is the first report on the impact of trans-bilayer asymmetry on the area expansion modulus of synthetic bilayer membranes. Since the mechanical properties of bilayer membranes play an important role in numerous cellular processes, these results have significant implications for membrane biology studies.

The effect of stabilizer on the mechanical response of double-emulsion-templated polymersomes.

Recent studies have shown that polymersomes templated by microfluidic double-emulsion possess several advantages such as high monodispersity and encapsulation efficiency compared with those generated based on thin-film rehydration and electroformation. Stabilizers, including bovine serum albumin (BSA) and polyvinyl alcohol (PVA), have been used to enhance the formation and stability of double emulsions that are used as templates for the generation of polymersomes. In this work, the effect of stabilizers on the mechanical response of double-emulsion-templated polymersomes using micropipette aspiration is investigated. It is demonstrated that the existence of stabilizers results in the inelastic response in poly-mersomes in the early stage of solvent removal. However, aged polymersomes that have little residual solvent show elastic behavior. Polymersomes prepared from PVA-stabilized double emulsions have noticeably lower area expansion moduli than polymersomes prepared from stabilizer-free and BSA-stabilized double emulsions, suggesting that PVA is incorporated in the bilayer membrane of polymersomes.

Optical Measurement of Biomechanical Properties of Human Red Blood Cell using Digital Holographic Microscopy: Malaria and Sickle Cell Diseases

Pathophysiological aspect of several hematologic diseases is largely determined by biomechanical properties of red blood cells (RBCs) and their hemodynamical properties in circulatory system 1. Here we present the biomechanical properties of individual RBCs from patients with sickle cell disease and RBCs infected with malaria-inducing parasites Pf. falciparum. using laser digital holographic technique, we non-invasively quantify membrane fluctuation in RBCs at the nanometer and millisecond scale, which is analyzed with the mathematical model to retrieve four important mechanical properties of RBCs; bending modulus, shear modulus, area expansion modulus, and cytoplasmic viscosity 2. We find significant alterations in the mechanical properties of RBCs in several pathophysiological states, ranging from depletion of Adenosine-5'-triphosphate (ATP) 3, different osmotic pressures 4, malaria infections 5-7, and sickle cell diseases 8,9.

Direct Measurements of Effect of Counterion Concentration on Mechanical Properties of Cationic Vesicles

Theoretical analyses of charged membranes in aqueous solutions have long predicted that the electric double layer surrounding them contributes significantly to their mechanical properties. Here we report the first, direct experimental measurements of the effect of counterion concentration on the bending and area expansion modulus of cationic surfactant vesicles. Using the classical technique of micropipet aspiration coupled with a modified experimental protocol that is better suited for cationic vesicles, we successfully measure the mechanical properties of a double-tailed cationic surfactant, diethylesterdimethyl ammonium chloride (diC18:1 DEEDMAC) in CaCl2 solutions. It is observed that the area expansion modulus of the charged membrane exhibits no measurable dependence on the counterion concentration, in accordance with existing models of bilayer elasticity. The measured bending modulus, however, is found to vary nonmonotonically and exhibits a minimum in its variation with counterion concentration. The experimental results are interpreted based on theoretical calculations of charged and bare membrane mechanics. It is determined that the initial decrease in bending modulus with increasing counterion concentration may be attributed to a decreasing double layer thickness, while the subsequent increase is likely due to an increasing membrane thickness. These mechanical moduli measurements qualitatively confirm, for the first time, theoretical predictions of a nonmonotonic behavior and the opposing effects of ionic strength on the bending rigidity of charged bilayers.

Optical Measurement of Biomechanical Properties of Human Red Blood Cell using Digital Holographic Microscopy: Malaria and Sickle Cell Diseases

Pathophysiological aspect of several hematologic diseases is largely determined by biomechanical properties of red blood cells (RBCs) and their hemodynamical properties in circulatory system 1. Here we present the biomechanical properties of individual RBCs from patients with sickle cell disease and RBCs infected with malaria-inducing parasites Pf. falciparum. using laser digital holographic technique, we non-invasively quantify membrane fluctuation in RBCs at the nanometer and millisecond scale, which is analyzed with the mathematical model to retrieve four important mechanical properties of RBCs; bending modulus, shear modulus, area expansion modulus, and cytoplasmic viscosity 2. We find significant alterations in the mechanical properties of RBCs in several pathophysiological states, ranging from depletion of Adenosine-5'-triphosphate (ATP) 3, different osmotic pressures 4, malaria infections 5-7, and sickle cell diseases 8,9.

Optical measurement of biomechanical properties of individual erythrocytes from a sickle cell patient

Sickle cell disease (SCD) is characterized by the abnormal deformation of red blood cells (RBCs) in the deoxygenated condition, as their elongated shape leads to compromised circulation. The pathophysiology of SCD is influenced by both the biomechanical properties of RBCs and their hemodynamic properties in the microvasculature. A major challenge in the study of SCD involves accurate characterization of the biomechanical properties of individual RBCs with minimum sample perturbation. Here we report the biomechanical properties of individual RBCs from a SCD patient using a non-invasive laser interferometric technique. We optically measure the dynamic membrane fluctuations of RBCs. The measurements are analyzed with a previously validated membrane model to retrieve key mechanical properties of the cells: bending modulus; shear modulus; area expansion modulus; and cytoplasmic viscosity. We find that high cytoplasmic viscosity at ambient oxygen concentration is principally responsible for the significantly decreased dynamic membrane fluctuations in RBCs with SCD, and that the mechanical properties of the membrane cortex of irreversibly sickled cells (ISCs) are different from those of the other types of RBCs in SCD.

Bursting Bubbles and Bilayers

This paper discusses various interactions between ultrasound, phospholipid monolayer-coated gas bubbles, phospholipid bilayer vesicles, and cells. The paper begins with a review of microbubble physics models, developed to describe microbubble dynamic behavior in the presence of ultrasound, and follows this with a discussion of how such models can be used to predict inertial cavitation profiles. Predicted sensitivities of inertial cavitation to changes in the values of membrane properties, including surface tension, surface dilatational viscosity, and area expansion modulus, indicate that area expansion modulus exerts the greatest relative influence on inertial cavitation. Accordingly, the theoretical dependence of area expansion modulus on chemical composition - in particular, poly (ethylene glyclol) (PEG) - is reviewed, and predictions of inertial cavitation for different PEG molecular weights and compositions are compared with experiment. Noteworthy is the predicted dependence, or lack thereof, of inertial cavitation on PEG molecular weight and mole fraction. Specifically, inertial cavitation is predicted to be independent of PEG molecular weight and mole fraction in the so-called mushroom regime. In the “brush” regime, however, inertial cavitation is predicted to increase with PEG mole fraction but to decrease (to the inverse 3/5 power) with PEG molecular weight. While excellent agreement between experiment and theory can be achieved, it is shown that the calculated inertial cavitation profiles depend strongly on the criterion used to predict inertial cavitation. This is followed by a discussion of nesting microbubbles inside the aqueous core of microcapsules and how this significantly increases the inertial cavitation threshold. Nesting thus offers a means for avoiding unwanted inertial cavitation and cell death during imaging and other applications such as sonoporation. A review of putative sonoporation mechanisms is then presented, including those involving microbubbles to deliver cargo into a cell, and those - not necessarily involving microubbles - to release cargo from a phospholipid vesicle (or reverse sonoporation). It is shown that the rate of (reverse) sonoporation from liposomes correlates with phospholipid bilayer phase behavior, liquid-disordered phases giving appreciably faster release than liquid-ordered phases. Moreover, liquid-disordered phases exhibit evidence of two release mechanisms, which are described well mathematically by enhanced diffusion (possibly via dilation of membrane phospholipids) and irreversible membrane disruption, whereas liquid-ordered phases are described by a single mechanism, which has yet to be positively identified. The ability to tune release kinetics with bilayer composition makes reverse sonoporation of phospholipid vesicles a promising methodology for controlled drug delivery. Moreover, nesting of microbubbles inside vesicles constitutes a truly “theranostic” vehicle, one that can be used for both long-lasting, safe imaging and for controlled drug delivery.

Soft biodegradable polymersomes from caprolactone-derived polymers

Polymersomes have promise for advanced theranostic delivery. We report here the design and characterization of a series of block copolymers that assemble into soft, bioresorbable, and non-toxic vesicles. The polymers are based on poly(ethylene glycol)–poly(caprolactone), but the caprolactone (CL) is copolymerized with a second monomer, 1,4,8-trioxaspiro-[4,6]-9-undecanone (TOSUO). Because TOSUO polymers have no crystalline character, copolymerizing TOSUO with CL should reduce the crystallinity of the polymersomes. After synthesizing polymers with different ratios of CL to TOSUO, we found that all copolymers assemble into both micron and nano-metric vesicles. Increasing the TOSUO content of the copolymer reduces the polymer crystalline melting temperature and the area expansion modulus of vesicle membranes. Membranes with partial crystalline structure exhibit hysteresis in the tension versus strain curve during aspiration. Vesicles are not cytotoxic and exhibit first-order release of encapsulated gemcitabine. These materials are promising for the development of deformable, biodegradable polymersomes for biomedical applications.