Live encapsulated Lactobacillus acidophilus cells in yogurt for therapeutic oral delivery: preparation and in vitro analysis of alginate–chitosan microcapsulesThis article is one of a selection of papers published in this special issue (part 1 of 2) on the Safety and Efficacy of Natural Health Products.

2007 ◽  
Vol 85 (9) ◽  
pp. 884-893 ◽  
Author(s):  
Aleksandra Malgorzata Urbanska ◽  
Jasmine Bhathena ◽  
Satya Prakash
Keyword(s):  
Lactobacillus Acidophilus ◽  
Targeted Delivery ◽  
Milk Fat ◽  
Oral Delivery ◽  
Live Cells ◽  
Natural Health Products ◽  
Bacterial Cells ◽  
Natural Polymers ◽  
Artificial Cells

Targeted delivery of live microencapsulated bacterial cells has strong potential for application in treating various diseases, including diarrhea, kidney failure, liver failure, and high cholesterol, among others. This study investigates the potential of microcapsules composed of two natural polymers, alginate and chitosan (AC), and the use of these artificial cells in yogurt for delivery of probiotic Lactobacillus acidophilus bacterial live cells. Results show that the integrity of AC microcapsules was preserved after 76 h of mechanical shaking in MRS broth and after 12 h and 24 h in simulated gastric and intestinal fluids. Using an in vitro computer-controlled simulated human gastrointestinal (GI) model, we found 8.37 log CFU/mL of viable bacterial cells were present after 120 min of gastric exposure and 7.96 log CFU/mL after 360 min of intestinal exposure. In addition, AC microcapsules composed of chitosan 10 and 100 at various concentrations were subjected to 4-week storage in 2% milk fat yogurt or 0.85% physiological solution. It was found that 9.37 log CFU/mL of cells encapsulated with chitosan 10 and 8.24 log CFU/mL of cells encapsulated with chitosan 100 were alive after 4 weeks. The AC capsule composed of 0.5% chitosan 10 provided the highest bacterial survival of 9.11 log CFU/mL after 4 weeks. Finally, an investigation of bacterial viability over 72 h in different pH buffers yielded highest survival of 6.34 log CFU/mL and 10.34 log CFU/mL at pH 8 for free and AC-encapsulated cells, respectively. We conclude from these findings that encapsulation allows delivery of a higher number of bacteria to desired targets in the GI tract and that microcapsules containing bacterial cells are good candidates for oral artificial cells for bacterial cell therapy.

scholarly journals Artificial Cell Therapy: New Strategies for the Therapeutic Delivery of Live Bacteria

2005 ◽  
Vol 2005 (1) ◽  
pp. 44-56 ◽  
Author(s):  
Satya Prakash ◽  
Mitchell Lawrence Jones
Keyword(s):  
Oral Delivery ◽  
Therapeutic Potential ◽  
Immobilized Cells ◽  
Genetically Engineered ◽  
Live Cells ◽  
Bacterial Cells ◽  
Artificial Cells ◽  
Artificial Cell ◽  
Wide Range ◽  
Live Bacterial Cells

There has been rapid growth in research regarding the use of live bacterial cells for therapeutic purposes. The recognition that these cells can be genetically engineered to synthesize products that have therapeutic potential has generated considerable interest and excitement among clinicians and health professionals. It is expected that a wide range of disease modifying substrates such as enzymes, hormones, antibodies, vaccines, and other genetic products will be used successfully and will impact upon health care substantially. However, a major limitation in the use of these bacterial cells is the complexity of delivering them to the correct target tissues. Oral delivery of live cells, lyophilized cells, and immobilized cells has been attempted but with limited success. Primarily, this is because bacterial cells are incapable of surviving passage through the gastrointestinal tract. In many occasions, when given orally, these cells have been found to provoke immunogenic responses that are undesirable. Recent studies show that these problems can be overcome by delivering live bacterial cells, such as genetically engineered cells, using artificial cell microcapsules. This review summarizes recent advances in the therapeutic use of live bacterial cells for therapy, discusses the principles of using artificial cells for the oral delivery of bacterial cells, outlines methods for preparing suitable artificial cells for this purpose, addresses potentials and limitations for their application in therapy, and provides insight for the future direction of this emergent and highly prospective technology.

scholarly journals Thermodynamics of protein destabilization in live cells

2015 ◽  
Vol 112 (40) ◽  
pp. 12402-12407 ◽  
Author(s):  
Jens Danielsson ◽  
Xin Mu ◽  
Lisa Lang ◽  
Huabing Wang ◽  
Andres Binolfi ◽  
...  
Keyword(s):  
Self Organization ◽  
Live Cells ◽  
Heat Capacity Change ◽  
Bacterial Cells ◽  
Crowding Effects ◽  
Capacity Change ◽  
The Individual ◽  
Cellular Components

Although protein folding and stability have been well explored under simplified conditions in vitro, it is yet unclear how these basic self-organization events are modulated by the crowded interior of live cells. To find out, we use here in-cell NMR to follow at atomic resolution the thermal unfolding of a β-barrel protein inside mammalian and bacterial cells. Challenging the view from in vitro crowding effects, we find that the cells destabilize the protein at 37 °C but with a conspicuous twist: While the melting temperature goes down the cold unfolding moves into the physiological regime, coupled to an augmented heat-capacity change. The effect seems induced by transient, sequence-specific, interactions with the cellular components, acting preferentially on the unfolded ensemble. This points to a model where the in vivo influence on protein behavior is case specific, determined by the individual protein’s interplay with the functionally optimized “interaction landscape” of the cellular interior.

scholarly journals Artificial cell microcapsules containing live bacterial cells and activated charcoal for managing renal failure creatinine: preparation and in-vitro analysis

2019 ◽  
Vol 3 (4) ◽  
pp. 190-196
Author(s):  
Trisna Lim ◽  
Wei Ouyang ◽  
Christopher John Martoni ◽  
Nasri Balit ◽  
Satya Prakash
Keyword(s):  
Lactobacillus Acidophilus ◽  
Activated Charcoal ◽  
Bacterial Cells ◽  
Removal Capacity ◽  
Artificial Cell ◽  
Urea Uptake ◽  
Vitro Analysis ◽  
Live Bacterial Cells ◽  
In Vitro Analysis

Abstract Activated charcoal was microencapsulated with Lactobacillus acidophilus 314 previously adapted for urea uptake. The creatinine removal capacity of this combination microcapsule was evaluated in-vitro in media simulating the small intestine. Results show that microcapsules containing both activated charcoal and L. acidophilus 314 demonstrated potential for decreasing creatinine. Interestingly, when co-encapsulating both activated charcoal and L. acidophilus 314 a smaller decrease in creatinine was observed than when encapsulating them separately. However, co-encapsulated microcapsules were more stable in various parts of the gastrointestinal system and survived longer in storage. These results suggest the feasibility of using microcapsules containing activated charcoal and probiotic bacteria as oral adjuvants for creatinine removal and provides a theoretical model for the use of these microcapsules to remove any unwanted metabolite.

scholarly journals Transcription-dependent confined diffusion of enzymes within subcellular spaces of the bacterial cytoplasm

BMC Biology ◽  
2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Daniel A. O. Rotter ◽  
Christoph Heger ◽  
Luis M. Oviedo-Bocanegra ◽  
Peter L. Graumann
Keyword(s):  
Single Cells ◽  
Single Particle Tracking ◽  
Live Cells ◽  
Fast Diffusion ◽  
Bacterial Cells ◽  
Substrate Channeling ◽  
Active Transcription ◽  
Bacterial Cytoplasm

Abstract Background Knowledge on the localization and mobility of enzymes inside bacterial cells is scarce, but important for understanding spatial regulation of metabolism. The four central enzymes (Rib enzymes) of the riboflavin (RF) biosynthesis pathway in the Gram positive model bacterium Bacillus subtilis have been studied extensively in vitro, especially the heavy RF synthase, a large protein complex with a capsid structure formed by RibH and an encapsulated RibE homotrimer, which mediates substrate-channeling. However, little is known about the behavior and mobility of these enzymes in vivo. Results We have investigated the localization and diffusion of the Rib enzymes in the cytoplasm of B. subtilis. By characterizing the diffusion of Rib enzymes in live cells using single particle tracking (SPT) we provide evidence for confined diffusion at the cell poles and otherwise Brownian motion. A majority of RibH particles showed clear nucleoid occlusion and a high degree of confined motion, which is largely abolished after treatment with Rifampicin, revealing that confinement is dependent on active transcription. Contrarily, RibE is mostly diffusive within the cell, showing only 14% encapsulation by RibH nanocompartments. By localizing different diffusive populations within single cells, we find that fast diffusion occurs mostly across the nucleoids located in the cell centers, while the slower, confined subdiffusion occurs at the crowded cell poles. Conclusions Our results provide evidence for locally different motion of active enzymes within the bacterial cytoplasm, setting up metabolic compartmentalization mostly at the poles of cells.

scholarly journals Investigation of Genipin Cross-Linked Microcapsule for Oral Delivery of Live Bacterial Cells and Other Biotherapeutics: Preparation and In Vitro Analysis in Simulated Human Gastrointestinal Model

2010 ◽  
Vol 2010 ◽  
pp. 1-10 ◽  
Author(s):  
Hongmei Chen ◽  
Wei Ouyang ◽  
Christopher Martoni ◽  
Fatemeh Afkhami ◽  
Bisi Lawuyi ◽  
...  
Keyword(s):  
Oral Delivery ◽  
Oral Therapy ◽  
Model Material ◽  
Bacterial Cells ◽  
Gi Tract ◽  
Viable Cells ◽  
Vitro Analysis ◽  
Live Bacterial Cells ◽  
In Vitro Analysis

Oral therapy utilizing engineered microorganisms has shown promise in the treatment of many diseases. By microencapsulation, viable cells can overcome the harsh gastrointestinal (GI) environment and secrete needed therapeutics into the gut. These engineered cells should be encased without escaping into the GI tract for safety concerns, thus robust microcapsule membrane is requisite. This paper examined the GI performance of a novel microcapsule membrane using a dynamic simulated human GI model. Results showed that the genipin cross-linked alginate-chitosan (GCAC) microcapsules possessed strong resistance to structural disintegration in the simulated GI environment. Leakage of encapsulated high molecular weight dextran, a model material to be protected during the simulated GI transit, was negligible over 72 h of exposure, in contrast to considerable leakage of dextran from the non-cross-linked counterparts. These microcapsules did not alter the microflora and enzymatic activities in the simulated human colonic media. This study suggested the potential of the GCAC microcapsules for oral delivery of live microorganisms and other biotherapeutics.

scholarly journals Preparation andIn VitroRelease of Drug-Loaded Microparticles for Oral Delivery Using Wholegrain Sorghum Kafirin Protein

2015 ◽  
Vol 2015 ◽  
pp. 1-8 ◽  
Author(s):  
Esther T. L. Lau ◽  
Stuart K. Johnson ◽  
Roger A. Stanley ◽  
Deirdre Mikkelsen ◽  
Zhongxiang Fang ◽  
...  
Keyword(s):  
Drug Delivery ◽  
Gastrointestinal Tract ◽  
Drug Delivery System ◽  
Delivery System ◽  
Targeted Delivery ◽  
Oral Delivery ◽  
Protein Digestibility ◽  
Target Area ◽  
Sorghum Grains

Kafirin microparticles have been proposed as an oral nutraceutical and drug delivery system. This study investigates microparticles formed with kafirin extracted from white and raw versus cooked red sorghum grains as an oral delivery system. Targeted delivery to the colon would be beneficial for medication such as prednisolone, which is used in the management of inflammatory bowel disease. Therefore, prednisolone was loaded into microparticles of kafirin from the different sources using phase separation. Differences were observed in the protein content,in vitroprotein digestibility, and protein electrophoretic profile of the various sources of sorghum grains, kafirin extracts, and kafirin microparticles. For all of the formulations, the majority of the loaded prednisolone was not released inin vitroconditions simulating the upper gastrointestinal tract, indicating that most of the encapsulated drug could reach the target area of the lower gastrointestinal tract. This suggests that these kafirin microparticles may have potential as a colon-targeted nutraceutical and drug delivery system.

scholarly journals Formulation Development of a Food-Graded Curcumin-Loaded Medium Chain Triglycerides-Encapsulated Kappa Carrageenan (CUR-MCT-KC) Gel Bead Based Oral Delivery Formulation

Materials ◽  
2021 ◽  
Vol 14 (11) ◽  
pp. 2783
Author(s):  
Kei-Xian Tan ◽  
Ling-Ling Evelyn Ng ◽  
Say Chye Joachim Loo
Keyword(s):  
Delivery System ◽  
Oral Delivery ◽  
Drug Carrier ◽  
In Vitro Release ◽  
Beta Carotene ◽  
Natural Polymers ◽  
Formulation Development ◽  
New Research ◽  
Kappa Carrageenan

In recent years, curcumin has been a major research endeavor in food and biopharmaceutical industries owing to its miscellaneous health benefits. There is an increasing amount of research ongoing in the development of an ideal curcumin delivery system to resolve its limitations and further enhance its solubility, bioavailability and bioactivity. The emergence of food-graded materials and natural polymers has elicited new research interests into enhanced pharmaceutical delivery due to their unique properties as delivery carriers. The current study is to develop a natural and food-graded drug carrier with food-derived MCT oil and a seaweed-extracted polymer called k-carrageenan for oral delivery of curcumin with improved solubility, high gastric resistance, and high encapsulation of curcumin. The application of k-carrageenan as a structuring agent that gelatinizes o/w emulsion is rarely reported and there is so far no MCT-KC system established for the delivery of hydrophobic/lipophilic molecules. This article reports the synthesis and a series of in vitro bio-physicochemical studies to examine the performance of CUR-MCT-KC as an oral delivery system. The solubility of CUR was increased significantly using MCT with a good encapsulation efficiency of 73.98 ± 1.57% and a loading capacity of 1.32 ± 0.03 mg CUR/mL MCT. CUR was successfully loaded in MCT-KC, which was confirmed using FTIR and SEM with good storage and thermal stability. Dissolution study indicated that the solubility of CUR was enhanced two-fold using heated MCT oil as compared to naked or unformulated CUR. In vitro release study revealed that encapsulated CUR was protected from premature burst under simulated gastric environment and released drastically in simulated intestinal condition. The CUR release was active at intestinal pH with the cumulative release of >90% CUR after 5 h incubation, which is the desired outcome for CUR absorption under human intestinal conditions. A similar release profile was also obtained when CUR was replaced with beta-carotene molecules. Hence, the reported findings demonstrate the potencies of MCT-KC as a promising delivery carrier for hydrophobic candidates such as CUR.

scholarly journals Development of a Novel Oral Delivery Vehicle for Probiotics

2020 ◽  
Vol 26 (26) ◽  
pp. 3134-3140 ◽  
Author(s):  
Kevin Enck ◽  
Surya Banks ◽  
Hariom Yadav ◽  
Mark E. Welker ◽  
Emmanuel C. Opara
Keyword(s):  
Targeted Delivery ◽  
Oral Delivery ◽  
Oral Drug Delivery ◽  
Oral Drug ◽  
Bacterial Cells ◽  
Alginate Hydrogel ◽  
Chemically Modified ◽  
Delivery Platform ◽  
Ph Conditions

Background: There is a significant interest in effective oral drug delivery of therapeutic substances. For probiotics, there is a particular need for a delivery platform that protects the bacteria from destruction by the acidic stomach while enabling targeted delivery to the intestine where microbiota naturally reside. The use of probiotics and how they impact the gut microbiota is a growing field and holds promise for the treatment of a variety of gastrointestinal diseases, including irritable bowel disease Crohn’s disease and C. diff and other diseases, such as obesity, diabetes, Parkinson’s, and Alzheimer’s diseases. Objective: The aim of this research was to use our newly developed chemically-modified alginate hydrogel with the characteristic feature of stability in acidic environments but disintegration under neutral-basic pH conditions to design a novel system for effective targeted delivery of ingested probiotics. Method and Results: We have used the approach of encapsulation of bacterial cells in the hydrogel of the modified alginate with in vitro studies in both simulated stomach acid and intestinal fluid conditions to demonstrate the potential application of this novel platform in oral delivery of probiotics. Our data provide a proof-of-concept that enables further studies in vivo with this delivery platform. Conclusion: We have demonstrated in the present study that our chemically modified alginate hydrogel is resistant to acidic conditions and protects bacterial cells encapsulated in it, but it is sensitive to neutral-basic pH conditions under which it disintegrates and releases its viable bacteria cell payload. Our data provide a proof-ofconcept that enables further studies in vivo with this delivery platform for the efficacy of therapeutic bacteria in various disease conditions.

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