Microbead Diameter Research Articles

Effect of high‐speed homogenisation on the morphological characteristics and survivability ofLactobacillus rhamnosusGG‐coated microbeads

SummaryXanthan gum can be utilised to overcome the structural defects in alginate microbeads and to improve the stability of microbeads under harsh conditions, that is, high acidic pH and homogenisation speeds. Present study was conducted to assess the survival ofLactobacillus rhamnosusGG using alginate (1.5% w/v) and xanthan gum (0.5% w/v) for encapsulation and structural modifications of microbeads. Homogenisation emulsion technique was used for encapsulation at various homogenisation speeds, that is, 3000 rpm (HS1), 6000 rpm (HS2), 9000 rpm (HS3) and 12 000 rpm (HS4). The resultant microbeads were characterised for diameter, beads yield, structural morphology using the scanning electron microscopy, probiotic count, bacterial survival, storage stability and homogenisation efficiency. Scanning electron microscopy images indicated that the beads produced at 12 000 rpm are more uniform, spherical in shape and have large surface area when compared with the other treatment groups. The diameter of microbeads was reduced from 323.1 to 238.2 μm as the homogenisation speed increased from 3000 to 12 000 rpm. The highest bacterial count (i.e., 9.45 log CFU g−1), homogenisation efficiency (91.6%), beads yield (23.0%), survivability (86.5%) and storage stability (9.84 to 7.90 log CFU g−1) were recorded at 12 000 rpm (HS4). Conclusively, the homogenised encapsulated microbeads at 12 000 rpm significantly improved the beads surface and sphericity, and bacterial survivability.

Injectable cell-laden hybrid bioactive scaffold containing bioactive glass microspheres.

The rising incidence of bone disorders has resulted in the need for minimally invasive therapies to meet this demand. Injectable bioactive filler, alone or with cells, could be applied in a minimally invasive manner to fulfill irregular cavities in non-load bearing sites, which do not require high mechanical properties. Thermosensitive chitosan hydrogels that transition from a liquid to a mechanically stable solid at body temperature provide interesting features as in-situ injectable cytocompatible biomaterials, but they are not osteoconductive. Osteoconductivity can be applied in combination with bioactive ceramics e.g., 45S5-Bioglass® (BG). However, BG addition in chitosan hydrogels results in pH elevation, due to rapid ions release, which adversely affects gel formation, mechanical properties, and cytocompatibility. To address this, we created hybrid hydrogels, where BG is concentrated in chitosan-based microbeads, incorporated in in-situ gelling chitosan hydrogels. We then compared the hybrid hydrogels' properties to chitosan hydrogels with homogenously distributed BG. By varying the stirred emulsification process, BG percentage, and CH formulation, we could tune the microbeads' properties. Incorporation of BG microbeads drastically improved the hydrogel's compressive modulus in comparison to homogeneously distributed BG. It also strongly increased the survival and metabolic activities of encapsulated cells. Calcium/phosphate increase on BG microbeads suggests hydroxyapatite formation. The small diameter of microbeads allows minimally invasive injection through small needles. The feasibility of freezing and thawing microbeads provides the possibility of long-term storage for potential clinical applications. These data indicate that this hybrid hydrogel forms a promising injectable cell-laden bioactive biomaterial for the treatment of unloaded bone defects.

Microfabrication of re-entrant surface with hydrophobicity/oleophobicity for liquid foods

Re-entrant texturing may potentially improve the hydrophobicity and oleophobicity of a surface. The food industry requires a microfabrication method to keep surfaces clean without leaving a packaging residue for applications such as food bottles, food containers, and preservation bags. The goal of this study is thus to establish a microfabrication method for re-entrant texturing with spherical curvature to produce hydrophobic/oleophobic surfaces for liquid foods, such as soy sauce and canola oil. Samples with a spherical curvature are created from an ultra-violet-cure (UV-cure) resin and poly (tetrafluoroethylene) (PTFE) microbeads with diameters between 2.26 to 1,353 microns by spin coating on a glass substrate. The resin thickness, the mass and diameter of the microbeads, and the spin coater rotation speed are used as the microfabrication parameters. A side view of samples showing the spherical curvature reveals that a re-entrant texture indeed forms. Distilled water, soy sauce, and canola oil are dropped softly onto the re-entrant surface, however, the droplets cannot be placed stably. For appropriate microbead diameters, the apparent contact angles of soy sauce and canola oil showed 130.2 and 119.4 degrees, respectively. This facile fabrication method for re-entrant surfaces could prove useful for generating hydrophobic/oleophobic surfaces for Newtonian liquid foods.

X-ray CT in Phase Contrast Enhancement Geometry of Alginate Microbeads in a Whole-Animal Model.

Imaging soft biomaterials in vivo is a significant challenge, as most conventional techniques are limited by biomaterial contrast, penetration depth, or spatial resolution. Exogeneous contrast agents used to increase contrast may also alter material properties or exhibit local toxicity. The capability to observe biomaterial constructs in vivo without introducing exogenous contrast would improve preclinical testing and evaluation. Conventional X-ray Computed Tomography allows fast, high-resolution imaging at high penetration depth, but biomaterial contrast is low. Previous studies employing X-ray phase contrast (XPC) and utilizing a synchrotron source provided support for the significant potential of XPC in imaging biomaterials without contrast agents. In this study, XPC tomography was used to image alginate hydrogel microspheres within a small animal omental pouch model using a commercially available X-ray source. Multilayer microbeads could be identified in the XPC images with volumetric and structural information not possible in histological analysis. The number of microbeads present and microbead volume and diameter could be quantified from the images. The results of this study show that XPC tomography can be a useful tool for monitoring of implanted soft biomaterials in small animal models.

Repeated detection of polystyrene microbeads in the Lower Rhine River

Microplastics are emerging pollutants in water bodies worldwide. The environmental entry areas must be studied to localise their sources and develop preventative and remedial solutions. Rivers are major contributors to the marine microplastics load. Here, we focus on a specific type of plastic microbead (diameter 286–954 μm, predominantly opaque, white–beige) that was repeatedly identified in substantial numbers between kilometres 677 and 944 of the Rhine River, one of Europe's main waterways. Specifically, we aimed (i) to confirm the reported abrupt increase in microbead concentrations between the cities of Leverkusen and Duisburg and (ii) to assess the concentration gradient of these particles along this stretch at higher resolution. Furthermore, we set out (iii) to narrow down the putative entry stretch from 81.3 km, as reported in an earlier study, to less than 20 km according to our research design, and (iv) to identify the chemical composition of the particles and possibly reveal their original purpose. Surface water filtration (mesh: 300 μm, n = 9) at regular intervals along the focal river stretch indicated the concentration of these spherules increased from 0.05 to 8.3 particles m−3 over 20 km. This spot sampling approach was supported by nine suspended solid samples taken between 2014 and 2017, encompassing the river stretch between Leverkusen and Duisburg. Ninety-five percent of microbeads analysed (202/212) were chemically identified as crosslinked polystyrene-divinylbenzene (PS-DVB, 146/212) or polystyrene (PS, 56/212) via Raman or Fourier-transform infrared spectroscopy. Based on interpretation of polymer composition, surface structure, shape, size and colour, the PS(-DVB) microbeads are likely to be used ion-exchange resins, which are commonly applied in water softening and various industrial purification processes. The reported beads contribute considerably to the surface microplastic load of the Rhine River and their potential riverine entry area was geographically narrowed down.

Tailored and biodegradable poly(2-oxazoline) microbeads as 3D matrices for stem cell culture in regenerative therapies

We present the synthesis of hydrogel microbeads based on telechelic poly(2-oxazoline) (POx) crosslinkers and the methacrylate monomers (HEMA, METAC, SPMA) by inverse emulsion polymerization. While in batch experiments only irregular and ill-defined beads were obtained, the preparation in a microfluidic (MF) device resulted in highly defined hydrogel microbeads. Variation of the MF parameters allowed to control the microbead diameter from 50 to 500 μm. Microbead elasticity could be tuned from 2 to 20 kPa by the POx:monomer composition, the POx chain length, net charge of the hydrogel introduced via the monomer as well as by the organic content of the aqueous phase. The proliferations of human mesenchymal stem cells (hMSCs) on the microbeads were studied. While neutral, hydrophilic POx-PHEMA beads were bioinert, excessive colonization of hMSCs on charged POx-PMETAC and POx-PSPMA was observed. The number of proliferated cells scaled roughly linear with the METAC or SPMA comonomer content. Additional collagen I coating further improved the stem cell proliferation. Finally, a first POx-based system for the preparation of biodegradable hydrogel microcarriers is described and evaluated for stem cell culturing.

Ex vivo 3D osteocyte network construction with primary murine bone cells.

Osteocytes reside as three-dimensionally (3D) networked cells in the lacunocanalicular structure of bones and regulate bone and mineral homeostasis. Despite of their important regulatory roles, in vitro studies of osteocytes have been challenging because: (1) current cell lines do not sufficiently represent the phenotypic features of mature osteocytes and (2) primary cells rapidly differentiate to osteoblasts upon isolation. In this study, we used a 3D perfusion culture approach to: (1) construct the 3D cellular network of primary murine osteocytes by biomimetic assembly with microbeads and (2) reproduce ex vivo the phenotype of primary murine osteocytes, for the first time to our best knowledge. In order to enable 3D construction with a sufficient number of viable cells, we used a proliferated osteoblastic population of healthy cells outgrown from digested bone chips. The diameter of microbeads was controlled to: (1) distribute and entrap cells within the interstitial spaces between the microbeads and (2) maintain average cell-to-cell distance to be about 19 µm. The entrapped cells formed a 3D cellular network by extending and connecting their processes through openings between the microbeads. Also, with increasing culture time, the entrapped cells exhibited the characteristic gene expressions (SOST and FGF23) and nonproliferative behavior of mature osteocytes. In contrast, 2D-cultured cells continued their osteoblastic differentiation and proliferation. This 3D biomimetic approach is expected to provide a new means of: (1) studying flow-induced shear stress on the mechanotransduction function of primary osteocytes, (2) studying physiological functions of 3D-networked osteocytes with in vitro convenience, and (3) developing clinically relevant human bone disease models.

Microplastic pollution in St. Lawrence River sediments

Although widely detected in marine ecosystems, microplastic pollution has only recently been documented in freshwater environments, almost exclusively in surface waters. Here, we report microplastics (polyethylene microbeads, 0.40–2.16 mm diameter) in the sediments of the St. Lawrence River. We sampled 10 freshwater sites along a 320 km section from Lake St. Francis to Québec City by passing sediment collected from a benthic grab through a 500 μm sieve. Microbeads were discovered throughout this section, and their abundances varied by four orders of magnitude across sites. Median and mean (±1 SE) densities across sites were 52 microbeads·m−2 and 13 832 (±13 677) microbeads·m−2, respectively. The highest site density was 1.4 × 105 microbeads·m−2 (or 103 microbeads·L−1), which is similar in magnitude to microplastic concentrations found in the world’s most contaminated marine sediments. Mean diameter of microbeads was smaller at sites receiving municipal or industrial effluent (0.70 ± 0.01 mm) than at non-effluent sites (0.98 ± 0.01 mm), perhaps suggesting differential origins. Given the prevalence and locally high densities of microplastics in St. Lawrence River sediments, their ingestion by benthivorous fishes and macroinvertebrates warrants investigation.

Structure, MC3T3-E1 Cell Response, and Osseointegration of Macroporous Titanium Implants Covered by a Bioactive Microarc Oxidation Coating with Microporous Structure

Macroporous Ti with macropores of 50-400 μm size is prepared by sintering Ti microbeads with different diameters of 100, 200, 400, and 600 μm. Bioactive microarc oxidation (MAO) coatings with micropores of 2-5 μm size are prepared on the macroporous Ti. The MAO coatings are composed of a few TiO2 nanocrystals and lots of amorphous phases with Si, Ca, Ti, Na, and O elements. Compared to compact Ti, the MC3T3-E1 cell attachment is prolonged on macroporous Ti without and with MAO coatings; however, the cell proliferation number increases. These results are contributed to the effects of the space structure of macroporous Ti and the surface chemical feature and element dissolution of the MAO coatings during the cell culture. Macroporous Ti both without and with MAO coatings does not cause any adverse effects in vivo. The new bone grows well into the macropores and micropores of macroporous Ti with MAO coatings, showing good mechanical properties in vivo compared to Ti, MAO-treated Ti, and macroporous Ti because of its excellent osseointegration. Moreover, the MAO coatings not only show a high interface bonding strength with new bones but also connect well with macroporous Ti. Furthermore, the pushing out force for macroporous Ti with MAO coatings increases significantly with increasing microbead diameter.

Measurement of Spatiotemporal Intracellular Deformation of Cells Adhered to Collagen Matrix During Freezing of Biomaterials

Preservation of structural integrity inside cells and at cell-extracellular matrix (ECM) interfaces is a key challenge during freezing of biomaterials. Since the post-thaw functionality of cells depends on the extent of change in the cytoskeletal structure caused by complex cell-ECM adhesion, spatiotemporal deformation inside the cell was measured using a newly developed microbead-mediated particle tracking deformetry (PTD) technique using fibroblast-seeded dermal equivalents as a model tissue. Fibronectin-coated 500 nm diameter microbeads were internalized in cells, and the microbead-labeled cells were used to prepare engineered tissue with type I collagen matrices. After a 24 h incubation the engineered tissues were directionally frozen, and the cells were imaged during the process. The microbeads were tracked, and spatiotemporal deformation inside the cells was computed from the tracking data using the PTD method. Effects of particle size on the deformation measurement method were tested, and it was found that microbeads represent cell deformation to acceptable accuracy. The results showed complex spatiotemporal deformation patterns in the cells. Large deformation in the cells and detachments of cells from the ECM were observed. At the cellular scale, variable directionality of the deformation was found in contrast to the one-dimensional deformation pattern observed at the tissue scale, as found from earlier studies. In summary, this method can quantify the spatiotemporal deformation in cells and can be correlated to the freezing-induced change in the structure of cytosplasm and of the cell-ECM interface. As a broader application, this method may be used to compute deformation of cells in the ECM environment for physiological processes, namely cell migration, stem cell differentiation, vasculogenesis, and cancer metastasis, which have relevance to quantify mechanotransduction.

Simultaneous determination of position and mass in the cantilever sensor using transfer function method

We present the simultaneous measurement of mass and position of micro-beads attached to the cantilever-based mass sensors using the transfer function method. 10 μm diameter micro-beads were placed on micro-cantilevers and the cantilevers were excited by lead-zirconate-titanate through low-pass filtered random voltages. The cantilever vibration was measured via a laser Doppler vibrometer before and after applying the beads. From the measured transfer function, the bead position was identified using its influence on the cantilever kinetic energy. The bead mass was then obtained by analyzing the wave propagation near the beads. The predicted position and mass agreed well with actual values.

Winner for outstanding research in the Ph.D. category for the 2013 society for biomaterials meeting and exposition, April 10–13, 2013, Boston, Massachusetts

Modular tissue engineering applies biomaterials-based approaches to create discrete cell-seeded microenvironments, which can be further assembled into larger constructs for the repair of injured tissues. In the current study, we embedded human bone marrow-derived mesenchymal stem cells (MSC) and human adipose-derived stem cells (ASC) in collagen/fibrin (COL/FIB) and collagen/fibrin/hydroxyapatite (COL/FIB/HA) microbeads, and evaluated their suitability for bone tissue engineering applications. Microbeads were fabricated using a water-in-oil emulsification process, resulting in an average microbead diameter of approximately 130 ± 25 μm. Microbeads supported both cell viability and cell spreading of MSC and ASC over 7 days in culture. The embedded cells also began to remodel and compact the microbead matrix as demonstrated by confocal reflectance microscopy imaging. After two weeks of culture in media containing osteogenic supplements, both MSC and ASC deposited calcium mineral in COL/FIB microbeads, but not in COL/FIB/HA microbeads. There were no significant differences between MSC and ASC in any of the assays examined, suggesting that either cell type may be an appropriate cell source for orthopedic applications. This study has implications in the creation of defined microenvironments for bone repair, and in developing a modular approach for delivery of pre-differentiated cells.

Video-based and interference-free axial force detection and analysis for optical tweezers

For measuring the minute forces exerted on single molecules during controlled translocation through nanopores with sub-piconewton precision, we have developed a video-based axial force detection and analysis system for optical tweezers. Since our detection system is equipped with a standard and versatile CCD video camera with a limited bandwidth offering operation at moderate light illumination with minimal sample heating, we integrated Allan variance analysis for trap stiffness calibration. Upon manipulating a microbead in the vicinity of a weakly reflecting surface with simultaneous axial force detection, interference effects have to be considered and minimized. We measured and analyzed the backscattering light properties of polystyrene and silica microbeads with different diameters and propose distinct and optimized experimental configurations (microbead material and diameter) for minimal light backscattering and virtually interference-free microbead position detection. As a proof of principle, we investigated the nanopore threading forces of a single dsDNA strand attached to a microbead with an overall force resolution of ±0.5 pN at a sample rate of 123 Hz.

Development of Na-CN—κ-carrageenan Microbeads for the Encapsulation of Lipophilic Compounds

The ionotropic gelation of double-layered emulsions composed of sodium caseinate and κ-carrageenan at pH values of 7 and 3.5 was evaluated, in order to obtain potential encapsulation matrices for hydrophobic compounds. The influence of some of the extrusion process variables (nozzle diameter at fluid exit and collecting distance) on the microbead production was studied, as well as the stability of the microbeads. The fluid nozzle diameter showed little influence on the shape of the microbeads, with a slight tendency for a decrease in microbead diameter with increase in fluid nozzle diameter. On the other hand, the collecting distance strongly influenced the microbead shape and they became more spherical (aspect ratio was reduced from ~2.0 to ~1.4) as the collecting distance was increased from 10 cm to 50 cm. The emulsion pH did not affect the aspect ratio of the microbeads, but the diameter was greater for microbeads produced at pH 3.5. This difference was attributed to the kind of interactions occurring between the κ-carrageenan and sodium caseinate at these distinct pH values. The microbeads were highly unstable when dispersed in deionized water, sugar solutions and low salt concentrations, releasing the encapsulated oil. However, no release of oil from the microbeads was observed when they were dispersed in ethanol or potassium chloride solutions with concentrations above 0.75 %, although their shape was modified when dispersed in ethanol. In general, the results obtained demonstrated the viability of the extrusion process to produce biopolymer-based microbeads and the potential application of these systems.

Laser direct-write of single microbeads into spatially-ordered patterns

Fabrication of heterogeneous microbead patterns on a bead-by-bead basis promotes new opportunities for sensors, lab-on-a-chip technology and cell-culturing systems within the context of customizable constructs. Laser direct-write (LDW) was utilized to target and deposit solid polystyrene and stem cell-laden alginate hydrogel beads into computer-programmed patterns. We successfully demonstrated single-bead printing resolution and fabricated spatially-ordered patterns of microbeads. The probability of successful microbead transfer from the ribbon surface increased from 0 to 80% with decreasing diameter of 600 to 45 µm, respectively. Direct-written microbeads retained spatial pattern registry, even after 10 min of ultrasonication treatment. SEM imaging confirmed immobilization of microbeads. Viability of cells encapsulated in transferred hydrogel microbeads achieved 37 ± 11% immediately after the transfer process, whereas randomly-patterned pipetted control beads achieved a viability of 51 ± 25%. Individual placement of >10 µm diameter microbeads onto planar surfaces has previously been unattainable. We have demonstrated LDW as a valuable tool for the patterning of single, micrometer-diameter beads into spatially-ordered patterns.

Construction and operation of a microrobot based on magnetotactic bacteria in a microfluidic chip.

Magnetotactic bacteria (MTB) are capable of swimming along magnetic field lines. This unique feature renders them suitable in the development of magnetic-guided, auto-propelled microrobots to serve in target molecule separation and detection, drug delivery, or target cell screening in a microfluidic chip. The biotechnology to couple these bacteria with functional loads to form microrobots is the critical point in its application. Although an immunoreaction approach to attach functional loads to intact MTB was suggested, details on its realization were hardly mentioned. In the current paper, MTB-microrobots were constructed by attaching 2 μm diameter microbeads to marine magnetotactic ovoid MO-1 cells through immunoreactions. These microrobots were controlled using a special control and tracking system. Experimental results prove that the attachment efficiency can be improved to ∼30% via an immunoreaction. The motility of the bacteria attached with different number of loads was also assessed. The results show that MTB can transport one load at a velocity of ∼21 μm/s and still move and survive for over 30 min. The control and tracking system is fully capable of directing and monitoring the movement of the MTB-microrobots. The rotating magnetic fields can stop the microrobots by trapping them as they swim within a circular field with a controllable size. The system has potential use in chemical analyses and medical diagnoses using biochips as well as in nano/microscale transport.

SSA-MOA: a novel CTC isolation platform using selective size amplification (SSA) and a multi-obstacle architecture (MOA) filter

Circulating tumor cells (CTCs) have gained increasing attention as physicians and scientists learn more about the role these extraordinarily rare cells play in metastatic cancer. In developing CTC technology, the critical criteria are high recovery rates and high purity. Current isolation methods suffer from an inherent trade-off between these two goals. Moreover, ensuring minimal cell stress and robust reproducibility is also important for the clinical application of CTCs. In this paper, we introduce a novel CTC isolation technology using selective size amplification (SSA) for target cells and a multi-obstacle architecture (MOA) filter to overcome this trade-off, improving both recovery rate and purity. We also demonstrate SSA-MOA's advantages in minimizing cell deformation during filter transit, resulting in more stable and robust CTC isolation. In this technique, polymer microbeads conjugated with anti-epithelial cell adhesion molecules (anti-EpCAM) were used to selectively size-amplify MCF-7 breast cancer cells, definitively differentiating from the white blood cells (WBCs) by avoiding the size overlap that compromises other size selection methods. 3 μm was determined to be the optimal microbead diameter, not only for size discrimination but also in maximizing CTC surface coverage. A multi-obstacle architecture filter was fabricated using silicon-on-glass (SOG) technology-a first such application of this fabrication technique-to create a precise microfilter structure with a high aspect ratio. The filter was designed to minimize cell deformation as simulation results predicted that cells captured via this MOA filter would experience 22% less moving force than with a single-obstacle architecture. This was verified by experiments, as we observed reliable cell capture and reduced cell deformation, with a 92% average recovery rate and 351 peripheral blood leukocytes (PBL) per millilitre (average). We expect the SSA-MOA platform to optimize CTC recovery rates, purity, and stability, increasing the sensitivity and reliability of such tests, thereby potentially expanding the utilization of CTC technologies in the clinic.

Field-deployable and near-real-time optical microfluidic biosensors for single-oocyst-level detection of Cryptosporidium parvum from field water samples

Cryptosporidium spp. is an obligate, parasitic protozoan that is difficult to detect and causes diarrhea in healthy adults while potentially causing death in the immunocompromised and children. Its treatment options are few and treat the symptoms, not the actual parasite. Current methods of detection are inefficient and rely too heavily upon laboratory sample preparations and technician skill, including differential staining, negative staining, and immunofluorescence methods [especially U.S. Environmental Protection Agency (EPA) Method 1623]. These assays can take from hours to days and require a laboratory environment. In this work, we demonstrated the microbead immunoagglutination assay combined with Mie scatter detection in a microfluidic device to provide a field-deployable and near-real-time alternative to the laboratory-based method (especially EPA Method 1623). Two main challenges were the relatively big diameter of Cryptosporidium oocysts (5-6 μm) and the contaminants in field water samples that negatively affected the immunoagglutination and its scatter detection. We used 4 min sonication to liberate Cryptosporidium oocyst wall proteins (COWP), which was previously used to inactivate Cryptosporidium oocysts. As for the contaminants, we optimized the microbead diameter (920 nm) and the wavelength of incident light (375 nm) to find the angle of scatter detection (45°) where the Mie scatter from immunoagglutinated microbeads was maximum and the background scatter from contaminants was minimum. This enabled the sub-single-oocyst-level detection despite the fact that only a very small volume of water sample (15 μL) was introduced to the microfluidic biosensor. When combined with filtration/concentration, this method is able to detect ≤1 oocyst per large volume of water, comparable to or potentially better than the EPA method 1623, while effectively reducing the time and labor necessary for staining and microscopic analysis. For faster, near-real-time assays, filtration/concentration may not be used, where the detection limit was 1-10 oocysts per mL with the total assay time of 10 min including the 4 min sonication time. The linear range of assay was over 5 orders of magnitude. The final device was compact and had the potential to be used in field situations, and required less technical expertise and/or training compared to the other methods.

The Application of High Pressure Electrostatic in Microcapsule Preparation

High pressure electrostatic (HPE) was applied to prepare microcapsules, and the operation parameters of HPE were investigated in this paper. When ecletrocity voltage is 20 kV and the distance between two electrodes is 5 cm, the diameter of micro-beads can be around 500 μm that is the optimal diameter for dispersing in water. The release rate of BSA was coming up to 45% after 120 min in AGF and even to 89% after 240 min in AIF at 37°C to basically meet the release in full. The amount of bacteria in 2% (w/v) Na-alginate microparticles declined to 4.08 log cfu per ml which is 4 times more than that of free bifidobacteria after 120 min in AGF with incubation time by 5 log cycles.

Phase-separated chitosan–fibrin microbeads for cell delivery

Matrix-enhanced delivery of cells is a promising approach to improving current cell therapies. Our objective was to create cell-laden composite microbeads that combine the attractive features of the natural polymers chitosan and fibrin. Liquid polydimethylsiloxane was used to emulsify a chitosan–fibrinogen solution containing suspended human fibroblast cells, followed by initiation of thrombin-mediated polymerization of fibrin and thermal/pH-mediated gelation of chitosan. Chitosan/fibrin weight percent (wt%) ratios of 100/0, 75/25, 50/50 and 25/75 were investigated. Microbead diameters ranged from 275 ± 99 µm to 38 ± 10 µm using impeller speeds from 600 to 1400 rpm. Fibroblasts remained viable on day 1 post-fabrication in all matrices, but cell viability was markedly higher in high-fibrin microbeads by day 8 post-fabrication. Cell spreading and interaction with the extracellular matrix was also markedly increased in high-fibrin matrices. Such composite microbeads containing viable entrapped cells have potential for minimally invasive delivery of cells for a variety of tissue repair applications.