Production and Mass Transfer Characteristics of Non-Newtonian Biopolymers for Biomedical Applications

ABSTRACT:?

The market for microbial biopolymers is currently expanding to include several emerging biomedical applications. Specifically, these applications are drug delivery and wound healing. A fundamental understanding of the key fermentation parameters is necessary in order to optimize the production of these biopolymers. Considering that most microbial biopolymer systems exhibit non-Newtonian rheology, oxygen mass transfer can be an important parameter to optimize and control. In this article, we present a critical review of recent advances in rheological and mass transfer characteristics of selected biopolymers of commercial interest in biomedical applications.     

Applications of biopolymers and other biotechnological products in building materials

Bio admixtures are functional molecules used in building products to optimize material properties. They include natural or modified biopolymers, biotechnological and biodegradable products. Concrete and dry-mix mortars (e.g. wall plasters or tile adhesives) represent two major applications for bio admixtures. Examples of bio products used in concrete are lignosulfonate, sodium gluconate, pine root extract, protein hydrolysates and Welan gum; and in dry-mix mortar methyl hydroxypropyl cellulose, hydroxypropyl starch, guar gum, tartaric acid, casein, succinoglycan and Xanthan gum. In a number of applications, bio admixtures compete well with synthetic admixtures. Sometimes, they are indispensable in the formulation of certain building products. Their market share is expected to increase because of technological advances, particularly in the field of microbial biopolymers, and because of the growing trend to use naturally based or biodegradable products in building materials.     

Novel chitin and chitosan nanofibers in biomedical applications

Chitin and its deacetylated derivative, chitosan, are non-toxic, antibacterial, biodegradable and biocompatible biopolymers. Due to these properties, they are widely used for biomedical applications such as tissue engineering scaffolds, drug delivery, wound dressings, separation membranes and antibacterial coatings, stent coatings, and sensors. In the recent years, electrospinning has been found to be a novel technique to produce chitin and chitosan nanofibers. These nanofibers find novel applications in biomedical fields due to their high surface area and porosity. This article reviews the recent reports on the preparation, properties and biomedical applications of chitin and chitosan based nanofibers in detail.     

Film forming microbial biopolymers for commercial applications—A review

Abstract

Microorganisms synthesize intracellular, structural and extracellular polymers also referred to as biopolymers for their function and survival. These biopolymers play specific roles as energy reserve materials, protective agents, aid in cell functioning, the establishment of symbiosis, osmotic adaptation and support the microbial genera to function, adapt, multiply and survive efficiently under changing environmental conditions. Viscosifying, gelling and film forming properties of these have been exploited for specific significant applications in food and allied industries. Intensive research activities and recent achievements in relevant and important research fields of global interest regarding film forming microbial biopolymers is the subject of this review. Microbial polymers such as pullulan, kefiran, bacterial cellulose (BC), gellan and levan are placed under the category of exopolysaccharides (EPS) and have several other functional properties including film formation, which can be used for various applications in food and allied industries. In addition to EPS, innumerable bacterial genera are found to synthesis carbon energy reserves in their cells known as polyhydroxyalkanoates (PHAs), microbial polyesters, which can be extruded into films with excellent moisture and oxygen barrier properties. Blow moldable biopolymers like PHA along with polylactic acid (PLA) synthesized chemically in vitro using lactic acid (LA), which is produced by LA bacteria through fermentation, are projected as biodegradable polymers of the future for packaging applications. Designing and creating of new property based on requirements through controlled synthesis can lead to improvement in properties of existing polysaccharides and create novel biopolymers of great commercial interest and value for wider applications. Incorporation of antimicrobials such as bacteriocins or silver and copper nanoparticles can enhance the functionality of polymer films especially in food packaging applications either in the form of coatings or wrappings. Use of EPS in combinations to obtain desired properties can be evaluated to increase the application range. Controlled release of active compounds, bioactive protection and resistance to water can be investigated while developing new technologies to improve the film properties of active packaging and coatings. An holistic approach may be adopted in developing an economical and biodegradable packaging material with acceptable properties. An interdisciplinary approach with new innovations can lead to the development of new composites of these biopolymers to enhance the application range. This current review focuses on linking and consolidation of recent research activities on the production and applications of film forming microbial polymers like EPS, PHA and PLA for commercial applications.     

One-step synthesis of polyesters specialties for biomedical applications

Polyesters are widely used for biomedical applications such as drug delivery systems and resorbable implants. The degradation kinetic of these biopolymers can be tailored by the introduction of functional groups in their backbone, leading to a modification of their morphology and hydrophilicity. This is usually realized via long multistep reaction pathways. This contribution describes the emergence of one-step procedures for this purpose including enzymatic and Lewis acid catalyzed polycondensation as well as coordinative ring opening polymerization.     

Gellan gum

For decades microbial exopolysaccharides have been invaluable ingredients in the food industry, as well as having many attractive pharmaceutical and chemical applications. Gellan gum is a comparatively new gum elaborated by the Gram-negative bacterium Sphingomonas paucimobilis. Although its physico-chemical properties have been well characterized, the ecology and physiology of Sphingomonas, and the factors influencing the fermentation process for production of this gum have received much less attention. This review focuses on the metabolism and the enzymic activity of this bacterium, as well as the factors that influence gellan production, including process temperature, pH, stirring rate, oxygen transfer, and composition of the production medium. Potential strategies for improving the production process are discussed in the context of processes for the production of other microbial biopolymers, particularly exopolysaccharides. In addition, the importance and potential utility of gellan lyases in modification of gellan and in other applications is critically evaluated.     

Microbial diversity as a source of useful biopolymers

Conclusions The few examples presented here, taken from research performed in my laboratory during the last 15 years, provide additional evidence that bacteria are a rich source of highly specialized polymers, many of which have potential commercial applications. Although modern molecular genetics is a valuable tool for modifying and overproducing proteins, the wonderful diversity of the microbial world remains the major source for discovering new and useful biopolymers. More than ever, the rate-limiting steps in discovering these microbial materials are imagination and techniques for enriching and screening for microorganisms that produce the desired products.     

Rheology,oxygen transfer,and molecular weight characteristics of poly(glutamic acid) fermentation by Bacillus subtilis

Poly(glutamic acid) (PGA) is a water-soluble, biodegradable biopolymer that is produced by microbial fermentation. Recent research has shown that PGA can be used in drug delivery applications for the controlled release of paclitaxel (Taxol) in cancer treatment. A fundamental understanding of the key fermentation parameters is necessary to optimize the production and molecular weight characteristics of poly(glutamic acid) by Bacillus subtilis for paclitaxel and other applications of pharmaceuticals for controlled release. Because of its high molecular weight, PGA fermentation broths exhibit non-Newtonian rheology. In this article we present experimental results on the batch fermentation kinetics of PGA production, mass transfer of oxygen, specific oxygen uptake rate, broth rheology, and molecular weight characterization of the PGA biopolymer.     

Recent developments in the synthesis of poly(hydroxybutyrate) based biocomposites

Poly(hydroxybutyrate) (PHB) has become an attractive biomaterial in research and development for past few years. It is natural bio-based aliphatic polyester produced by many types of bacteria. Due to its biodegradable, biocompatible, and eco-friendly nature, PHB can be used in line with bioactive species. However, high production cost, thermal instability, and poor mechanical properties limit its desirable applications. So there is need to incorporate PHB with other materials or biopolymers for the development of some novel PHB based biocomposites for value addition. Many attempts have been employed to incorporate PHB with other biomaterials (or biopolymers) to develop sustainable biocomposites. In this review, some recent developments in the synthesis of PHB based biocomposites and their biomedical, packaging and tissue engineering applications have been focused. The development of biodegradable PHB based biocomposites with improved mechanical properties could be used to overcome its native limitations hence to open new possibilities for industrial applications.     

Fabrication of three-dimensional collagen scaffold using an inverse mould-leaching process

Natural biopolymers, such as collagen or chitosan, are considered ideal for biomedical scaffolds. However, low processability of the materials has hindered the fabrication of designed pore structures controlled by various solid freeform-fabrication methods. A new technique to fabricate a biomedical three-dimensional collagen scaffold, supplemented with a sacrificial poly(ethylene oxide) mould is proposed. The fabricated collagen scaffold shows a highly porous surface and a three-dimensional structure with high porosity as well as mechanically stable structure. To show its feasibility for biomedical applications, fibroblasts/keratinocytes were co-cultured on the scaffold, and the cell proliferation and cell migration of the scaffold was more favorable than that obtained with a spongy-type collagen scaffold.     

Production of chondroitin sulfate and chondroitin

The production of microbial polysaccharides has recently gained much interest because of their potential biotechnological applications. Several pathogenic bacteria are known to produce capsular polysaccharides, which provide a protection barrier towards harsh environmental conditions, and towards host defences in case of invasive infections. These capsules are often composed of glycosaminoglycan-like polymers. Glycosaminoglycans are essential structural components of the mammalian extracellular matrix and they have several applications in the medical, veterinary, pharmaceutical and cosmetic field because of their peculiar properties. Most of the commercially available glycosaminoglycans have so far been extracted from animal sources, and therefore the structural similarity of microbial capsular polysaccharides to these biomolecules makes these bacteria ideal candidates as non-animal sources of glycosaminoglycan-derived products. One example is hyaluronic acid which was formerly extracted from hen crests, but is nowadays produced via Streptococci fermentations. On the other hand, no large scale biotechnological production processes for heparin and chondrotin sulfate have been developed. The larger demand of these biopolymers compared to hyaluronic acid (tons vs kilograms), due to the higher titre in the final product (grams vs milligrams/dose), and the scarce scientific effort have hampered the successful development of fermentative processes. In this paper we present an overview of the diverse applications and production methods of chondroitin reported so far in literature with a specific focus on novel microbial biotechnological approaches.     

Sustainable films and coatings from hemicelluloses: a review

This review summarizes the results of past research on films and coatings from hemicelluloses, biopolymers that are as yet relatively unexploited commercially. The targeted uses of hemicelluloses have primarily been packaging films and coatings for foodstuffs as well as biomedical applications. Oxygen permeability of hemicellulose films, an important characteristic for food packaging, was typically comparable to values found for other biopolymer films such as amylose and amylopectin. As expected, the modification of hemicelluloses to create more hydrophobic films reduced the water vapor permeability. However, modified hemicellulose coatings intended for food still exhibited water vapor permeabilities several magnitudes higher than those of other polymers currently used for this purpose. Research on hemicelluloses for biomedical applications has included biocompatible hydrogels and coatings and material surfaces with enhanced cell affinity. Numerous possibilities exist for chemically modifying hemicelluloses, and fundamental studies of films from modified hemicelluloses have identified other potential applications, including selective membranes.     

Mixing,mass transfer,and scale-up of polysaccharide fermentations

Microbial polysaccharides are rapidly emerging as a new and important source of polymeric materials. These biopolymers have novel and unique properties and already have found a wide range of applications in the food, pharmaceutical, and other industries. In view of the impending importance of polysaccharides as an industrial commodity, there is renewed interest in the area of product and process development. This paper summarizes the state-of-the art in polysaccharide fermentations. An attempt is being made to review the following areas: rheological characteristics of polysaccharide solutions, mixing and power requirements of polysaccharides and other highly viscous non-Newtonian systems, oxygen mass transfer, and scale-up problems encountered in polysaccharide fermentations.     

Polylactic acid: synthesis and biomedical applications

Social and economic development has driven considerable scientific and engineering efforts on the discovery, development and utilization of polymers. Polylactic acid (PLA) is one of the most promising biopolymers as it can be produced from nontoxic renewable feedstock. PLA has emerged as an important polymeric material for biomedical applications on account of its properties such as biocompatibility, biodegradability, mechanical strength and process ability. Lactic acid (LA) can be obtained by fermentation of sugars derived from renewable resources such as corn and sugarcane. PLA is thus an eco-friendly nontoxic polymer with features that permit use in the human body. Although PLA has a wide spectrum of applications, there are certain limitations such as slow degradation rate, hydrophobicity and low impact toughness associated with its use. Blending PLA with other polymers offers convenient options to improve associated properties or to generate novel PLA polymers/blends for target applications. A variety of PLA blends have been explored for various biomedical applications such as drug delivery, implants, sutures and tissue engineering. PLA and their copolymers are becoming widely used in tissue engineering for function restoration of impaired tissues due to their excellent biocompatibility and mechanical properties. The relationship between PLA material properties, manufacturing processes and development of products with desirable characteristics is described in this article. LA production, PLA synthesis and their applications in the biomedical field are also discussed.     

Affinity capillary electrophoresis: important application areas and some recent developments

Affinity capillary electrophoresis (ACE) is a broad term referring to the separation by capillary electrophoresis of substances that participate in specific or non-specific affinity interactions during electrophoresis. The interacting molecules can be found free in solution or can be immobilized to a solid support. Every ACE mode has advantages and disadvantages. Each can be used for a wide variety of applications. This paper focuses on applications that include purification and concentration of analytes present in diluted solutions or complex matrices, quantitation of analytes based on calibration curves, and estimation of binding constants from direct and derived binding curves based on quantitation of analytes or on analyte migration shifts. A more recent chemicoaffinity strategy in capillary electrophoresis/capillary electrochromatography (CE/CEC) termed molecular imprinting (`plastic antibodies') is discussed as well. Although most ACE studies are aimed at characterizing small-molecular mass analytes such as drugs, hormones, and peptides, some efforts have been pursued to characterize larger biopolymers including proteins, such as immunoglobulins. Examples of affinity interactions that have been studied are antigen–antibody, hapten–antibody, lectin–sugar, drug–protein, and enzyme–substrate complexes using ultraviolet, laser-induced fluorescence, and mass spectrometer detectors. This paper also addresses the critical issue of background electrolyte selection and quantitation of analytes. Specific examples of bioaffinity applications are presented, and the future of ACE in the biomedical field is discussed.     

Elastin-like recombinamers: biosynthetic strategies and biotechnological applications

The past few decades have witnessed the development of novel naturally inspired biomimetic materials, such as polysaccharides and proteins. Likewise, the seemingly exponential evolution of genetic-engineering techniques and modern biotechnology has led to the emergence of advanced protein-based materials with multifunctional properties. This approach allows extraordinary control over the architecture of the polymer, and therefore, monodispersity, controlled physicochemical properties, and high sequence complexity that would otherwise be impossible to attain. Elastin-like recombinamers (ELRs) are emerging as some of the most prolific of these protein-based biopolymers. Indeed, their inherent properties, such as biocompatibility, smart nature, and mechanical qualities, make these recombinant polymers suitable for use in numerous biomedical and nanotechnology applications, such as tissue engineering, "smart" nanodevices, drug delivery, and protein purification. Herein, we present recent progress in the biotechnological applications of ELRs and the most important genetic engineering-based strategies used in their biosynthesis.     

Absorption of a mixture of volatile organic compounds (VOCs) in aqueous solutions of soluble cutting oil

A semi-industrial bioscrubber was developed to treat a complex mixture of VOCs: oxygenated, aromatic and chlorinated compounds. In order to optimize the VOCs mass transfer, an original washing agent made up of water and cutting oil was tested, and the impact of this washing agent on bioscrubbing performances was investigated. The results obtained with a laboratory unit show that the addition of oil strongly increases the quantity of transferred aromatics. For these compounds, the apparent mass transfer coefficient k(L)a is lower than with water alone. In term of bioscrubbing performances, comparison of the results obtained with the water-oil mixture and water alone showed that the removal efficiency for aromatics is enhanced: from 12% to 36% (applied load of 852 g VOCs m(-3)h(-1)); the elimination of chlorinated compounds is slightly improved. The addition of oil does not seem to lead to any dysfunction of the microbial communities that metabolize the transferred compounds.     

The future prospects of microbial cellulose in biomedical applications

Microbial cellulose has proven to be a remarkably versatile biomaterial and can be used in wide variety of applied scientific endeavors, such as paper products, electronics, acoustics, and biomedical devices. In fact, biomedical devices recently have gained a significant amount of attention because of an increased interest in tissue-engineered products for both wound care and the regeneration of damaged or diseased organs. Due to its unique nanostructure and properties, microbial cellulose is a natural candidate for numerous medical and tissue-engineered applications. For example, a microbial cellulose membrane has been successfully used as a wound-healing device for severely damaged skin and as a small-diameter blood vessel replacement. The nonwoven ribbons of microbial cellulose microfibrils closely resemble the structure of native extracellular matrices, suggesting that it could function as a scaffold for the production of many tissue-engineered constructs. In addition, microbial cellulose membranes, having a unique nanostructure, could have many other uses in wound healing and regenerative medicine, such as guided tissue regeneration (GTR), periodontal treatments, or as a replacement for dura mater (a membrane that surrounds brain tissue). In effect, microbial cellulose could function as a scaffold material for the regeneration of a wide variety of tissues, showing that it could eventually become an excellent platform technology for medicine. If microbial cellulose can be successfully mass produced, it will eventually become a vital biomaterial and will be used in the creation of a wide variety of medical devices and consumer products.     

Production of glucuronic acid-based polysaccharides by microbial fermentation for biomedical applications

This review provides an overview of the properties, different biosynthetic machineries, and biotechnological production processes of four microbially derived glucuronic acid-based polysaccharides that are of interest for diverse biomedical purposes. In particular, the utilization of hyaluronic acid and heparin sulfate in high-value medical applications is already well established, whereas chondroitin sulfate and alginate show high potential within this ever-growing field. Furthermore, new strategies exploiting genetically engineered microorganisms generated through improving naturally existing pathways or de novo designed ones are described. These new developments result in increased fermentation titers, and thereby, pave the way towards feasible, or at least improved, process economy. Moreover, these strategies also allow for the future possibility of producing tailor-made biopolymers with specified characteristics, even novel molecules.     

元基因组学及其在转化医学中的应用

微生物群落遍布于人体的每个角落,与人共生并对人体健康产生重要和深刻的影响。与人类共生的全部微生物的基因组总和称为“元基因组”或“人类第二基因组”。研究人体微生物群落及相关元基因组数据,对转化医学领域的基础研究和临床应用具有重要的价值。通过对生物医学相关的高通量元基因组数据进行分析,不仅能为基础医学研究向医学临床应用转化提供新思路和新方法,而且具有广阔的应用前景。基于新一代测序技术产生的数据,元基因组分析技术和方法能够弥补以往人体微生物先培养后鉴定方法的缺陷,同时能有效鉴定和分析微生物群落的组成及功能,从而进一步探究和揭示微生物群落与机体生理状态之间的关系,为解决许多医学领域的难题提供了全新的切入角度和思维方法。文章系统介绍了元基因组研究的现状,包括元基因组的方法概念和研究进展,并以元基因组在医学研究中的应用为着眼点,综述了元基因组在转化医学方面的研究进展,进一步阐述了元基因组研究在转化医学应用领域中具有的重要地位。