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JOURNAL OF APPLIED POLYMER SCIENCE · 2017 ·Vol. 135 ·期 2 ·45685 · IF 1.86

Preparation and optimization of calcium pectate beads for cell encapsulation

Shan Zhao,Ying Zhang,Yang Liu,Fan Yang,Zhilong Xiu,Xiaojun Ma,Guangwei Sun

A variety of natural polymers are considered suitable for cell entrapment. Pectin, a natural polysaccharide found in the cell walls of all higher terrestrial plants, presents great potential for cell encapsulation because of its ability to form gels. In this study, we investigated the influences of intrinsic and extrinsic factors on the formation and properties of calcium pectate (CP). Beads of CP prepared with 30–35% methyl esterification presented improved gel properties. The molecular weight ( M w ), calcium content, and gel time did not significantly affect the gel properties. The gelling properties, sphericity factor (SF), and swelling ratio were significantly influenced by the degree of methyl esterification (DE) and pectin concentration. The DE (<40%), M w , and calcium concentration had negative effects on the SFs of CP beads. Moreover, increases in the DE (<40%) and pectin concentration increased the swelling ratio. The C3A cells encapsulated in optimized CP beads maintained good viability and proliferation for up to 2 weeks. In conclusion, CP beads are a potential encapsulation material for cell immobilization and application in related fields. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2018 , 135 , 45685.

研究领域: 生物材料学 · 细胞生物学
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CARBOHYDRATE POLYMERS · 2011 · IF 3.463

Thermo-sensitive alginate-based injectable hydrogel for tissue engineering

Rongwei Tan,Zhending She,Mingbo Wang,Zhou Fang,Yuansheng Liu,Qingling Feng

A thermo-sensitive comb-like copolymer was synthesized by grafting PNIPAAm-COOH with a single carboxy end group onto aminated alginate (AAlg) through amide bond linkages. In the copolymer, alginate was the backbone and poly(N-isopropylacrylamide) (PNIPAAm) was the pendant group. The structures of AAlg and three AAlg-g-PNIPAAm copolymers with different PNIPAAm grafting ratios were determined by FTIR and 1 H NMR. The rheological properties of AAlg-g-PNIPAAm copolymer hydrogels were measured by monitoring the viscosity, storage modulus and loss modulus as a function of temperature. The lower critical solution temperature of AAlg-g-PNIPAAm copolymers was measured as 35   °C through rheological analysis. An in vitro degradation study was carried out by monitoring weight loss. It was confirmed that degradation can be controlled by PNIPAAm modification. Encapsulation of human bone mesenchymal stem cells (hBMSCs) within hydrogels showed that the AAlg-g-PNIPAAm copolymer was not cytotoxic and preserved the viability of the entrapped cells well. The thermo-sensitive AAlg-g-PNIPAAm copolymer has attractive properties that make it suitable as cell or pharmaceutical delivery vehicles for a variety of tissue engineering applications.

研究领域: 生物材料学 · 组织工程学
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES · 2018 · IF 3.909

Optimization of preparation conditions for calcium pectinate with response surface methodology and its application for cell encapsulation

Shan Zhao,Ying Zhang,Yang Liu,Fan Yang,Wenguang Yu,Shixin Zhang,Xiaojun Ma,Guangwei Sun

The most notable and unique property of pectin is its ability to form gels, which is the foundation of its many functions and applications. To obtain the desired pectin hydrogel beads for tissue engineering or biological applications, the combined effect of the key factors of gel properties of calcium pectinate (CP) beads were investigated by response surface methodology (RSM). The results derived from RSM indicated that the model equation of average size and mechanical stability were significant and could be used to describe the process under a wide range of preparation conditions. The optimum condition for preparing variables were gained graphically. Moreover, the degree of methyl-esterification (DE) of pectin and pectin - calcium concentrates showed significant and combined effort on sphericity factor (SF). The mechanical stability of CP beads was significantly affected by the quadratic contributions of the pectin concentration, the interaction effects between pectin and calcium concentration, and the linear contribution of pectin concentration. The viability and proliferation of cells encapsulated in optimal CP beads demonstrated that the optimal formula results in better gel properties and are more suitable to cell encapsulation.

研究领域: 生物工程 · 材料科学
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JOURNAL OF CONTROLLED RELEASE · 2024 · IF 10.5

Encapsulation of individual mammalian cells as a cell-based drug delivery carrier for lung cancer treatment

Chaochen Shi,Tao Chen,Yingying Li,Wenshuai Li,Yuan Shen,Kehan Cai,Ming Wang,Yazhou Chen

Cell-based delivery holds great promise for advancing cell therapy, but it often faces challenges such as low cell survival rates and immunological rejection during cell injection and circulation. In this study, we develop an alternative approach aimed at engineering cell membranes to produce encapsulated individual mammalian (EIM) cells. The encapsulation shell was formed by catalyzing the reaction of H 2 O 2 with hyaluronic acid-dopamine (HA-DA) on the cell surface using horseradish peroxidase (HRP) enzymes. This protective shell not only protects live mammalian cells from physical stress but also from biological assaults. The individual cell encapsulation system enables the loading of cells with chemotherapy drugs, enhancing their targeting ability towards tumor sites and resulting in over 5.1-fold cell enrichment at metastatic tumors, thereby improving tumor-killing efficacy and reducing metastasis. Overall, the individual cell encapsulation system demonstrates potential for targeted drug delivery to the lungs in the treatment of lung cancer by reducing side effects and enhancing treatment outcomes.

CAS / MDL: 51-61-6
研究领域: 生物医学工程 · 肿瘤学 · 药理学
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Small · 2018 ·Vol. 14 ·期 36 ·1801095 · IF 9.598

A Microfluidic Strategy for Controllable Generation of Water-in-Water Droplets as Biocompatible Microcarriers

Hai-Tao Liu,Hui Wang,Wen-Bo Wei,Hui Liu,Lei Jiang,Jian-Hua Qin

Droplet microfluidics has been widely applied in functional microparticles fabricating, tissue engineering, and drug screening due to its high throughput and great controllability. However, most of the current droplet microfluidics are dependent on water-in-oil (W/O) systems, which involve organic reagents, thus limiting their broader biological applications. In this work, a new microfluidic strategy is described for controllable and high-throughput generation of monodispersed water-in-water (W/W) droplets. Solutions of polyethylene glycol and dextran are used as continuous and dispersed phases, respectively, without any organic reagents or surfactants. The size of W/W droplets can be precisely adjusted by changing the flow rate of dispersed and continuous phases and the valve switch cycle. In addition, uniform cell-laden microgels are fabricated by introducing the alginate component and rat pancreatic islet (β-TC6) cell suspension to the dispersed phase. The encapsulated islet cells retain high viability and the function of insulin secretion after cultivation for 7 days. The high-throughput droplet microfluidic system with high biocompatibility is stable, controllable, and flexible, which can boost various chemical and biological applications, such as bio-oriented microparticles synthesizing, microcarriers fabricating, tissue engineering, etc.

研究领域: 生物工程 · 材料科学 · 细胞生物学
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Soft Matter · 2012 ·Vol. 9 ·期 1 ·132-137 · IF 4.39

Fabrication of a shape memory hydrogel based on imidazole–zinc ion coordination for potential cell-encapsulating tubular scaffold application

Wenjing Nan,Wei Wang,Han Gao,Wenguang Liu

A dipole–dipole reinforced copolymer hydrogel was synthesized by the one-step photopolymerization of vinylimidazole (VI) and acrylonitrile (AN) comonomer in the presence of a polyethylene glycol-based crosslinker. The mechanical properties of PVI–AN gels were tremendously increased by only chelating several mmol L−1 zinc ions, which was shown to firmly lock the temporary shape of the gel. Extraction of zinc ions by a complexing agent could facilitate the recovery of a permanent shape, and the memory behavior was reversible. The PVI–AN hydrogels supported the growth of L929 cells. The cell-seeded flat hydrogel sheet was folded into a temporary tubular scaffold and fixed in a culture medium containing 5 mmol L−1 zinc ions. After unfolding the tube, the exposed L929 cells were shown to adhere well on the surface of the gel. It is anticipated that this cell-loaded SM hydrogel could be fixed to a desired shape in vitro by small numbers of zinc ions for a potential implantable tissue engineering scaffold.

CAS / MDL: 1072-63-5
研究领域: 生物材料学 · 组织工程学
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JOURNAL OF COLLOID AND INTERFACE SCIENCE · 2017 · IF 4.233

Antimicrobial colloidal hydrogels assembled by graphene oxide and thermo-sensitive nanogels for cell encapsulation

Wenhua Cheng,Yunhua Chen,Lijing Teng,Bingheng Lu,Li Ren,Yingjun Wang

Hydrogels are promising 3D materials that have demonstrated increasing applications in the encapsulation and delivery of drugs and cells. Herein we report an injectable colloidal hydrogel that directly assembled by graphene oxide (GO) and thermo-sensitive nanogels (tNG). The pH dependent hydrogen bonding interactions between the carboxyl and oxethyl groups induce the reversible assembly of GO and nanogels. The hydrogel is mouldable and can be shaped into different macroscopic objects, and the mechanical strengths are tunable with pH and temperature adjustment. The hybrid hydrogel by its own possesses high antibacterial activity , and demonstrates responsive drug release behaviour and high viability of 3D encapsulated cells. We expect this hybrid colloidal hydrogel can serve as an interesting scaffold for active cargo delivery and cell culture.

研究领域: 生物材料学 · 组织工程学 · 药剂学
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ANALYTICAL AND BIOANALYTICAL CHEMISTRY · 2023 · IF 4.3

Demand-driven active droplet generation and sorting based on positive pressure-controlled fluid wall

Zhang Yiwei,Lin Yiwei,Hong Xianzhe,Di Chao,Xin Yuelai,Wang Xinru,Qi Shuhong,Liu Bi-Feng,Zhang Zhihong,Du Wei

Droplet microfluidics is a rapidly advancing area of microfluidic technology, which offers numerous advantages for cell analysis, such as isolation and accumulation of signals, by confining cells within droplets. However, controlling cell numbers in droplets is challenging due to the uncertainty of random encapsulation which result in many empty droplets. Therefore, more precise control techniques are needed to achieve efficient encapsulation of cells within droplets. Here, an innovative microfluidic droplet manipulation platform had been developed, which employed positive pressure as a stable and controllable driving force for manipulating fluid within chips. The air cylinder, electro-pneumatics proportional valve, and the microfluidic chip were connected through a capillary, which enabled the formation of a fluid wall by creating a difference in hydrodynamic resistance between two fluid streams at the channel junction. Lowering the pressure of the driving oil phase eliminates hydrodynamic resistance and breaks the fluid wall. Regulating the duration of the fluid wall breakage controls the volume of the introduced fluid. Several important droplet microfluidic manipulations were demonstrated on this microfluidic platform, such as sorting of cells/droplets, sorting of droplets co-encapsulated cells and hydrogels, and active generation of droplets encapsulated with cells in a responsive manner. The simple, on-demand microfluidic platform was featured with high stability, good controllability, and compatibility with other droplet microfluidic technologies. Graphical abstract

CAS / MDL: 110-18-9
研究领域: 微流控 · 细胞分析 · 生物技术 · 芯片实验室
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ACS Applied Materials & Interfaces · 2016 · IF 7.145

In Situ “Clickable” Zwitterionic Starch-Based Hydrogel for 3D Cell Encapsulation

Dianyu Dong,Junjie Li,Man Cui,Jinmei Wang,Yuhang Zhou,Liu Luo,Yufei Wei,Lei Ye,Hong Sun,Fanglian Yao

Three-dimensional (3D) cell encapsulation in hydrogel provides superb methods to investigate the biochemical cues in directing cellular fate and behaviors outside the organism, the primary step of which is to establish suitable “blank platform” to mimic and simplify native ECM microenvironment. In this study, zwitterionic starch-based “clickable” hydrogels were fabricated via a “copper- and light- free” Michael-type “thiol–ene” addition reaction between acylated-modified sulfobetaine-derived starch (SB-ST-A) and dithiol-functionalized poly(ethylene glycol) (PEG-SH). By incorporating antifouling SB-ST and PEG, the hydrogel system would be excellently protected from nontarget protein adsorption to act as a “blank platform”. The hydrogels could rapidly gel under physiological conditions in less than 7 min. Dynamic rheology experiments suggested the stiffness of the hydrogel was close to the native tissues, and the mechanical properties as well as the gelation times and swelling behaviors could be easily tuned by varying the precursor proportions. The protein and cell adhesion assays demonstrated that the hydrogel surface could effectively resist nonspecific protein and cell adhesion. The degradation study in vitro confirmed that the hydrogel was biodegradable. A549 cells encapsulated in the hydrogel maintained high viability (up to 93%) and started to proliferate in number and extend in morphology after 2 days’ culture. These results indicated the hydrogel presented here could be a potential candidate as “blank platform” for 3D cell encapsulation and biochemical cues induced cellular behavior investigation in vitro.

CAS / MDL: 25322-68-3
研究领域: 生物材料学 · 组织工程学 · 细胞生物学
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LAB ON A CHIP · 2017 ·Vol. 17 ·期 19 ·3310-3317 · IF 6.045

Microfluidic generation of aqueous two-phase-system (ATPS) droplets by oil-droplet choppers

Chunmei Zhou,Pingan Zhu,Ye Tian,Xin Tang,Rui Shi,Liqiu Wang

Existing approaches for droplet generation with an ultra-low interfacial tension using aqueous two-phase systems, ATPS, are either constricted by a narrow range of flow conditions using passive methods or subjected to complex chip fabrication with the integration of external components using active actuation. To address these issues, we present a simple approach to produce uniform ATPS droplets facilitated by oil-droplet choppers in microfluidics. Our solution counts on the synchronized formation of high-interfacial-tension oil-in-water and low-interfacial-tension water-in-water droplets, where the ATPS interface is distorted by oil droplets and decays into water-in-water droplets. In the synchronization regime, the size and generation frequency of ATPS droplets can be controlled independently by tuning the flow rates of the dispersed aqueous and oil phases, respectively. Our method demonstrates high uniformity of droplets (coefficient of variation between 0.75% and 2.45%), a wide range of available droplet size (droplet radius from 5 μm to 180 μm), and a maximum generation frequency of about 2.1 kHz that is nearly two orders of magnitude faster than that in existing methods. We develop theoretical models to precisely predict the minimum and maximum frequencies of droplet generation and the droplet size. The produced ATPS droplets and oil choppers are separated in the channel using density difference. Our method would boost emulsion-based biological applications such as cell encapsulation, biomolecule delivery, bioreactors, and biomaterials synthesis with ATPS droplets.

研究领域: 生物医学工程 · 微流控技术 · 生物材料合成 · 细胞生物学 · 药物递送系统
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ACS Applied Bio Materials · 2023 · IF 4.7

Microfluidic Fabrication of Gelatin Acrylamide Microgels through Visible Light Photopolymerization for Cell Encapsulation

Tao Tang,Chang Liu,Zeqi Min,Wenjing Cai,Xiacong Zhang,Wen Li,Afang Zhang

Gelatin-based microgels are intriguing for various biomedical applications, which are conventionally prepared through photopolymerization of gelatin methacrylamide (GelMA). Here, we report on the modification of gelatin through acrylamidation to form gelatin acrylamide (GelA) with different substitution degrees, which was found to exhibit fast photopolymerization kinetics, better gelation, steady viscosity at elevated temperatures, and satisfying biocompatibility when compared to GelMA. By the online photopolymerization strategy with a home-made microfluidic setting, microgels of uniform sizes from GelA by blue light were obtained and their swollen properties were investigated. Compared to the microgels from GelMA, they showed an enhanced cross-linking degree and have better shape stability when swollen in water. Cell toxicities of the hydrogels from GelA and cell encapsulation from the corresponding microgels were investigated, which were found to exhibit superior properties than those from GelMA. We therefore believe that GelA has potential for constructing scaffolds for bioapplications and can be an excellent substitute for GelMA.

CAS / MDL: 106797-53-9 760-93-0
研究领域: 生物医学工程 · 材料科学 · 细胞生物学
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Acta Biomaterialia · 2020 · IF 7.242

Continuous microfluidic encapsulation of single mesenchymal stem cells using alginate microgels as injectable fillers for bone regeneration

Chuanfeng An,Weijian Liu,Yang Zhang,Bo Pang,Hui Liu,Yujie Zhang,Haoyue Zhang,Liyuan Zhang,Hongbing Liao,Changle Ren,Huanan Wang

The encapsulation of cells in microscale hydrogels can provide a mimic of a three-dimensional (3D) microenvironment to support cell viability and functions and to protect cells from the environmental stress, which have been widely used in tissue regeneration and cell therapies. Here, a microfluidics-based approach is developed for continuous encapsulation of mesenchymal stem cells (MSCs) at the single-cell level using alginate microgels. This microfluidic technique integrated on-chip encapsulation, gelation , and de-emulsification into a one-step fabrication process, which enables scalable cell encapsulation while retaining the viability and functionality of loaded cells. Remarkably, we observed MSCs encapsulated in Ca-alginate microgels at the single-cell level showed significantly enhanced osteogenesis and accelerated mineralization of the microgels which occurred only after 7 days of induction. Furthermore, MSCs laden in alginate microgels displayed significantly enhanced bone formation compared to MSCs mixed with microgels and acellular microgels in a rat tibial ablation model. To conclude, the current microfluidic technique represents a significant step toward continuous single cell encapsulation, fabrication, and purification . These microgels can boost bone regeneration by providing a controlled osteogenic microenvironment for encapsulated MSCs and facilitate stem cell therapy in the treatment of bone defects in a minimally invasive delivery way. Statement of Significance The biological functions and therapeutic activities of single cells laden in microgels for tissue engineering remains less investigated. Here, we reported a microfluidic-based method for continuous encapsulation of single MSCs with high viability and functionality by integrating on-chip encapsulation, gelation, and de-emulsification into a one-step fabrication process. More importantly, MSCs encapsulated in alginate microgels at the single-cell level showed significantly enhanced osteogenesis, remarkably accelerated mineralization in vitro and bone formation capacity in vivo . Therefore, this single-cell encapsulation technique can facilitate stem cell therapy for bone regeneration and be potentially used in a variety of tissue engineering applications.

研究领域: 组织工程 · 干细胞治疗 · 生物医学工程 · 再生医学
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