Structural transitions in TPGS micelles induced by trehalose as a model cryoprotectant

Arya, Shristi; Patidar, Rajesh Kumar; Ray, Debes; Aswal, Vinod K.; Ranjan, Nihar; Bahadur, Pratap; Tiwari, Sanjay

doi:10.1016/j.colsurfa.2022.128714

Cited by 8 publications

(8 citation statements)

References 39 publications

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“…These arguments are well supported by the data on the cryoprotectant factor. 4,12 The values were found to be 0.8 and 0.91 for SL and SL-CHL liposomes, respectively, before the incorporation of sugars. Interestingly, it increased to 1 upon adding sugars.…”

Section: Resultsmentioning

confidence: 96%

“…At higher concentrations, sugar dehydrates the membrane, probably by pulling water molecules toward itself. 12 This triggers interdigitation among phospholipid molecules, thus leading to minor changes in the structural parameters of vesicles. At a threshold concentration, a reduction in the lateral mobility of phospholipids leads to glassy state formation.…”

Section: Resultsmentioning

confidence: 99%

“…A number of sugars and polyols have been tested to preserve the integrity of liposomes during lyophilization. , Incorporation of mono-, di-, and oligo-saccharides is common. − In general, cryoprotectants form a glassy matrix in the bilayer at subzero temperature . It has been reported that sugars interact with the bilayer in a concentration-dependent manner.…”

Section: Introductionmentioning

confidence: 99%

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Sugar-based Cryoprotectants Stabilize Liposomal Vesicles by Exhibiting a Cholesterol-like Effect

Sikandar,

Kumar,

Rathod

et al. 2024

Mol. Pharmaceutics

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Liposomal vesicles tend to fuse and aggregate during lyophilization. To avoid these events, cryoprotectants are added to the dispersion before lyophilization. Herein, we have compared the effect of three commonly used cryoprotectants (mannitol, MTL; trehalose, THL; and β-cyclodextrin, β-CD) upon structural characteristics of liposomes. The formulation was prepared using ethanol injection method, and cryoprotectants were tested at three dose levels (2, 6, and 10 mM). We have elucidated their effect on soy lecithin (SL) liposomes formulated with and without cholesterol (CHL). Characterizations were performed using scattering, thermal, and spectroscopic techniques. CHL molecules interacted hydrophobically with the SL bilayer. In spite of triggering a noticeable increase in the hydrodynamic diameter (about 30 nm), CHL promoted the stabilization of vesicles. Hydrogen bonding interactions were verified by the shift in −OH stretching over 3300−3500 cm −1 . This manifested in an increased phase transition temperature (T m ) of SL liposomes. T m increased further upon incorporation of cryoprotectants, particularly with β-CD. Enthalpic changes were indicative of an affinity interaction between phospholipids and cryoprotectants, regardless of the presence of CHL. β-CD showed concentration-dependent changes in the energetics of this interaction. The affinity of cryoprotectant-liposome interaction has been ranked as β-CD ≫ THL > MNT.

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Section: Resultsmentioning

confidence: 96%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Sugar-based Cryoprotectants Stabilize Liposomal Vesicles by Exhibiting a Cholesterol-like Effect

Sikandar,

Kumar,

Rathod

et al. 2024

Mol. Pharmaceutics

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“…The process depends upon the type of additive and their concentration. Sugars, salts, and solvents have been shown to modulate the clouding through disruption of water structures or induction of growth of aggregates. , The difference in clouding between T1304 and T1307 can be clearly noticed. Given its high hydrophilicity (PEO: 70%), T1307 remained solubilized even at a higher temperature.…”

Section: Resultsmentioning

confidence: 99%

Comparative Effect of Physiological Salts upon Micellization of T1304 and T1307

et al. 2023

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We present a comprehensive investigation on the interaction of tetronics (T1304 and T1307) with some important physiological salts (NaH2PO4, KH2PO4, Na2CO3, NaCl, and KI). Thermodynamic and microstructural aspects of these interactions were studied as a function of the solution temperature, pH and salt concentration. Characterizations were performed using turbidimetric, calorimetric, and scattering techniques. We show that, at ambient temperature, T1304 molecules aggregated to form spherical core–shell aggregates displaying a unimodal distribution pattern. On the other hand, unimers and large clusters dominated in the case of highly hydrophilic T1307. Its micellization was promoted in the presence of salts as per the following trend: NaCl < KH2PO4 < NaH2PO4 ≪ Na2CO3. Aggregation was found to be endothermic, and hydrophobic interactions (TΔS mic > ΔH mic) prevailed. The enthalpy–entropy compensation plot was found to be linear for both copolymers. Demicellization occurred in the presence of KI as it facilitated the buildup of water structures around the copolymer chains. This could be verified from the increase in the cloud point, critical micelle concentration, and free energy. Overall, the temperature and salts inflicted a stronger hydrophobic effect upon T1304 in comparison to T1307.

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“…Strains preserved in the presence of trehalose demonstrated a consistent recovery time (days to fluoresce once rehydrated), with average recovery times between 2 and 4 days through 10 months of storage (Figure 3e). Beyond stabilizing cellular components, it is possible that trehalose may interact with F127‐BUM polymer architecture during freeze‐drying, as has been seen in the case of other micellular assemblies (Arya et al, 2022). These results directly contrast to those without an additive wherein the recovery time increased by 2–3 days at each storage timepoint tested (Figure 3b).…”

Section: Resultsmentioning

confidence: 99%

Enhancing long‐term storage and stability of engineered living materials through desiccant storage and trehalose treatment

Brooks

Reed

Yuan

et al. 2022

Biotech & Bioengineering

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Engineered living materials (ELMs) have broad applications for enabling on-demand bioproduction of compounds ranging from small molecules to large proteins. However, most formulations and reports lack the capacity for storage beyond a few months. In this study, we develop an optimized procedure to maximize stress resilience of yeast-laden ELMs through the use of desiccant storage and 10% trehalose incubation before lyophilization. This approach led to over 1-year room temperature storage stability across a range of strain genotypes. In particular, we highlight the superiority of exogenously added trehalose over endogenous, engineered production in yielding robust preservation resilience that is independent of cell state. This simple, effective protocol enables sufficient accumulation of intracellular trehalose over a short period of contact time across a range of strain backgrounds without requiring the overexpression of a trehalose importer. A variety of microscopic analysis including µ-CT and confocal microscopy indicate that cells form spherical colonies within F127-BUM ELMs that have variable viability upon storage. The robustness of the overall procedure developed here highlights the potential for widespread deployment to enable on-demand, cold-chain independent bioproduction.

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Structural transitions in TPGS micelles induced by trehalose as a model cryoprotectant

Cited by 8 publications

References 39 publications

Sugar-based Cryoprotectants Stabilize Liposomal Vesicles by Exhibiting a Cholesterol-like Effect

Sugar-based Cryoprotectants Stabilize Liposomal Vesicles by Exhibiting a Cholesterol-like Effect

Comparative Effect of Physiological Salts upon Micellization of T1304 and T1307

Enhancing long‐term storage and stability of engineered living materials through desiccant storage and trehalose treatment

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