Preparation and characterization of novel sinomenine microcapsules for oral controlled drug delivery

Background: Developing a sustained release drug to cure arthritis is needed. Sinomenine (SIN) is abstracted from sinomenium acutum and widely used in the treatment of various rheumatism and arrhythmia with few side effects. The primary aim of this study is to develop SIN microcapsules with polyelectrolyte multilayers for controlled drug release. Method: SIN microcrystals were encapsulated with chitosan, gelatin, and alginate by layer-by-layer technique, such as (gelatin/alginate)₄ and (chitosan/alginate)₆. The size distribution, zeta-potential, stability, and morphology of the microcapsules were characterized by a particle size analyzer, zetasizer, ultraviolet spectroscopy, and transmission electron microscope, respectively. The in vitro controlled release pattern of SIN was studied using a diffusion cell assembly at physiological pH of 6.8 or 1.4. Results: Light stability of these microcapsules was improved after microencapsulation. Compared with release rate of the SIN microcapsules coated by the poly(dimethyldiallyl ammonium chloride)/alginate and gelatin/alginate multilayers, release rate of the SIN microcapsules coated with chitosan/alginate multilayers was fast. Release rate progressively decreased with the increase of chitosan/alginate bilayer number and the decrease of pH value of release medium. Conclusion: These novel SIN microcapsules may be developed into oral controlled drug delivery for rheumatism and arthritis.     

Injectable calcium phosphate–alginate–chitosan microencapsulated MC3T3-E1 cell paste for bone tissue engineering in vivo

Osteoblasts or stem cells have been delivered into injectable calcium phosphate cement (CPC) to improve its effectiveness and biological function. However, the osteogenic potential of the new construct in vivo has been rarely reported, and there are no reports on alginate–chitosan microencapsulated osteoblasts mixed with CPC. This study aimed to develop alginate–chitosan microencapsulated mouse osteoblast MC3T3-E1 cells (AC-cells), evaluate the osteogenic potential of a calcium phosphate cement complex with these AC-cells (CPC-AC-cell), and trace the implanted MC3T3-E1 cells in vivo. MC3T3-E1 cells were embedded in alginate microcapsules, cultured in osteogenic medium for 7 days, and then covered with chitosan before mixing with a paste of β-tricalcium phosphate/calcium phosphate cement (β-TCP/CPC). The construct was injected into the dorsal subcutaneous area of nude mice. Lamellar-bone-like mineralization, newly formed collagen and angiogenesis were observed at 4 weeks. At 8 weeks, areas of newly formed collagen expanded; further absorption of β-TCP/CPC and osteoid-like structures could be seen. Cell tracing in vivo showed that implanted MC3T3-E1 cells were clearly visible at 2 weeks. These in vivo results indicate that the novel injectable CPC-AC-cell construct is promising for bone tissue engineering applications.     

Preparation of turmeric oil-loaded chitosan-alginate biopolymeric nanocapsules

Preparation of chitosan-alginate nanocapsules containing turmeric oil was carried out by emulsification of turmeric oil in aqueous sodium alginate solution and gelification with calcium chloride and chitosan, followed by solvent removal. The process parameters were optimized by variation of the molecular weight of chitosan, the chitosan/alginate mass ratio, and the order of addition of calcium chloride and chitosan in the formulation. The nanocapsules were characterized based on average size, zeta potential, morphology, percent recovery, loading capacity of turmeric oil, nanocapsule yield, and stability at 4 °C and 25 °C. The characteristics of the nanocapsules were dependent on the molecular weight and amount of chitosan. Chitosan with a low molecular weight was required to produce small nanocapsules. At a fixed chitosan/alginate mass ratio of 0.1:1, addition of chitosan after calcium chloride was found to be optimal for improving the physical stability, percent recovery of turmeric oil and nanocapsule yield, while maintaining the loading capacity of the turmeric oil.     

Preparation and Characterization of Coacervate Microcapsules for the Delivery of Antimicrobial Oyster Peptides

Oyster peptides-loaded alginate/chitosan/starch microcapsules were prepared using external gelation method and internal emulsion gelation method. The solution of oyster peptides complexes was encapsulated into the microcapsules, which endowed the microcapsules with intestine passive targeting properties. The swelling behavior, encapsulation efficiency, and release behavior of oyster peptides from the microcapsules at different pH values were investigated. The microcapsules exhibited sustained release of the peptides in intestinal medium, and the release rate could be regulated by the pH value: in simulated gastric fluid, the release rate was greatly decreased, and in simulated body fluid and intestinal fluid, the microcapsules exhibited a sustained release in 24 h with different release rates. The microspheres were characterized by Fourier transform infrared. The results suggested that the alginate/chitosan/starch microcapsules could be a suitable copolymeric carrier system for intestinal protein or peptides delivery in the intestine.     

Preparation and characterization of coacervate microcapsules for the delivery of antimicrobial oyster peptides

Oyster peptides-loaded alginate/chitosan/starch microcapsules were prepared using external gelation method and internal emulsion gelation method. The solution of oyster peptides complexes was encapsulated into the microcapsules, which endowed the microcapsules with intestine passive targeting properties. The swelling behavior, encapsulation efficiency, and release behavior of oyster peptides from the microcapsules at different pH values were investigated. The microcapsules exhibited sustained release of the peptides in intestinal medium, and the release rate could be regulated by the pH value: in simulated gastric fluid, the release rate was greatly decreased, and in simulated body fluid and intestinal fluid, the microcapsules exhibited a sustained release in 24 h with different release rates. The microspheres were characterized by Fourier transform infrared. The results suggested that the alginate/chitosan/starch microcapsules could be a suitable copolymeric carrier system for intestinal protein or peptides delivery in the intestine.     

Preparation of embolic NEMs loading capecitabine

The nanoparticles-embedded microcapsules (NEMs) with smooth surface, good sphericity, excellent dispersivity and uniform particle size distribution were prepared by emulsification combined with electrospraying to extend the sustained release performance of the embolic microcapsules loading capecitabine (CAP). The sodium alginate and chitosan with good biocompatibility were used as the materials and CAP as a small-molecule model drug. The drug loading, encapsulation efficiency and drug release of CAP in the NEMs were investigated. The results showed that the drug-loading and encapsulation efficiency both increased with the increment of chitosan and CAP concentration. The maximum values of drug loading and encapsulation efficiency were 1.97 and 18.01 % respectively when initial CAP concentration was 5.0 g/L and chitosan molecular weight 100 kDa. The cumulative release rate of CAP released from the NEMs was lower than 30 % in 0.5 h, which indicated that there was no obvious initial burst release behavior. In the subsequent 240 h, the release results confirmed that the NEMs had better sustained release properties compared to pure microcapsules, and it might be a new anticancer drug delivery system in the future studies.     

微胶囊化脂肪酶型TTI的制备及其性能研究

针对脂肪酶在时间-温度指示剂的应用中存在热稳定性能差,温度、pH值对其活性影响较大等问题,采用离子凝胶技术制备脂肪酶微胶囊。应用单因素试验和正交试验相结合的方法,以海藻酸钠、壳聚糖、氯化钙浓度为变量,优化脂肪酶微胶囊的制备工艺,对原脂肪酶与脂肪酶微胶囊的热稳定性能、最适温度与pH值进行研究,采用红外光谱（FTIR）、扫描电镜（SEM）对脂肪酶微胶囊结构及形貌进行分析,并对脂肪酶微胶囊在时间-温度指示剂中的应用做出初步研究。得到了脂肪酶微胶囊最佳制备工艺条件为：海藻酸钠质量浓度30 g/L,壳聚糖质量浓度6 g/L,氯化钙浓度0.3 mol/L。脂肪酶微胶囊的热稳定性比原脂肪酶有明显提高。脂肪酶微胶囊制备工艺稳定、可靠,具有应用于时间-温度指示剂中的实用价值。     

Encapsulation of Huh-7 cells within alginate-poly(ethylene glycol) hybrid microspheres

Novel calcium alginate poly(ethylene glycol) hybrid microspheres (Ca-alg-PEG) were developed and evaluated as potentially suitable materials for cell microencapsulation. Grafting 5–13% of the backbone units of sodium alginate (Na-alg) with α-amine-ω-thiol PEG maintained the gelling capacity in presence of calcium ions, while thiol end groups allowed for preparing chemically crosslinked hydrogel via spontaneous disulfide bond formation. The combination of these two gelling mechanisms yielded Ca-alg-PEG. Human hepatocellular carcinoma cells (Huh-7) were encapsulated in Ca-alg-PEG and calcium alginate beads (Ca-alg), and cultured for 2 weeks under agitation conditions. Immediately after completion of the microencapsulation, the cell viability was 60% and similar in Ca-alg-PEG and Ca-alg. The proliferation of Huh-7 encapsulated in Ca-alg-PEG was slightly higher than in Ca-alg. Accelerated proliferation after 2 weeks was observed for the encapsulation in Ca-alg-PEG. The production of albumin confirmed the functionality of the encapsulated Huh-7 cells. The study confirms the suitability of Ca-alg-PEG and the one-step technology for cell microencapsulation.     

肉桂精油微囊化及其在果蔬保鲜中的应用

目的以壳聚糖、海藻酸钠为壁材,以肉桂精油为芯材,采用复凝聚法制备壳聚糖微胶囊,并将其应用于水果保鲜中。方法通过光学显微镜、扫描电子显微镜、傅里叶转化红外光谱等,确定微胶囊的最佳工艺参数、芯壁比及添加质量分数,并以芒果为实验对象,研究添加不同微胶囊质量分数的壳聚糖溶液对芒果保鲜效果的影响。结果当壳聚糖/海藻酸钠质量比为1∶2,氯化钙质量分数为1%,p H值为5,戊二醛质量分数为60%,交联时间为60 min,芯壁比为4∶1时,制备形成的壳聚糖微胶囊成球效果较好;微胶囊在壳聚糖溶液中的添加质量分数为4%时,以其涂膜的芒果保鲜效果最佳。结论肉桂精油经囊化后有效减少了气味、延缓了挥发,在果蔬保鲜中有较重要的现实研究意义。     

The effect of alginate addition on the structure and morphology of hydroxyapatite/gelatin nanocomposites

A series of hydroxyapatite/gelatin/alginate nanocomposites with different amount of alginate were synthesized by a co-precipitation method. With the increase of alginate amount, a cross-linked alginate/gelatin polymer network formed, which induced a gradual red shift of organic absorption peaks in FT-IR analysis. TEM images indicated that the development of HAP nanocrystals in an aqueous gelatin/alginate mixture was highly influenced by the alginate content. On increasing alginate content, the dimensions of the crystals increased and their morphology changed from needle-like to long fiber-like, and at high alginate content, the crystals tended to aggregate in separate clusters. The results of the electron diffraction strongly indicated alginate promoted the preferential alignment in c direction of HAP nanocrystals. SEM results showed that high amount of alginate led to regular shape and large size of HAP crystals after the composites were calcined for 4 h at 600 °C.     

Formulation and Comparative Characterization of Chitosan,Gelatin, and Chitosan–Gelatin-Coated Liposomes of CPT-11–HCl

Liposomes containing phosphatidylcholine and cholesterol (uncoated) and coated by chitosan, gelatin, and combination of chitosan and gelatin were prepared by the modified ethanol injection method. The aim of this work was to formulate and characterize liposomes of camptothecin (CPT)-11–HCl (Irinotecan HCl) containing chitosan, gelatin, and both polymers as coating materials; and also to increase its circulation longevity when compared with the free drug while maintaining the agent in its active lactone form. Size, shape, zeta potential, encapsulation efficiency, stability study, in vitro, and in vivo release study were used for characterization of liposomes. The size of liposomes was in the order of uncoated?<?chitosan coated?<?gelatin coated?<?combination of chitosan and gelatin coated. The zeta potential of liposomes was in the order of combination of chitosan and gelatin coated?>?chitosan coated?>?gelatin coated?>?uncoated. The formulations showed the long-term stability. The encapsulation efficiency of liposomes was in order of combination of chitosan and gelatin coated?>?gelatin coated?>?chitosan coated?>?uncoated. The in vitro and in vivo release of drug was observed in the order of combination chitosan and gelatin coated?>?gelatin coated?>?chitosan coated?>?uncoated.     

Sustained-release alginate-chitosan pellets prepared by melt pelletization technique

Context: Alginate-chitosan pellets prepared by extrusion-spheronization technique exhibited fast drug dissolution.

Objective: This study aimed to design sustained-release alginate pellets through rapid in situ matrix coacervation by chitosan during dissolution.

Methods: Pellets made of alginate with chitosan and/or calcium acetate were prepared using solvent-free melt pelletization technique which prevented reaction between processing materials during agglomeration and allowed such reaction to occur only in dissolution phase.

Results: Drug release was retarded in pH 2.2 medium when pellets were formulated with calcium acetate or chitosan till a change in medium pH to 6.8. The sustained-release characteristics of calcium alginate pellets were attributed to pellet dispersion and rapid cross-linking by soluble Ca²⁺ during dissolution. The slow drug release characteristics of alginate-chitosan pellets were attributed to polyelectrolyte complexation and pellet aggregation into swollen structures with reduced erosion. The drug release was, however, not retarded when both calcium acetate and chitosan coexisted in the same matrix as a result of chitosan shielding of Ca²⁺ to initiate alginate cross-linkages and rapid in situ solvation of calcium acetate induced fast pellet dispersion and chitosan losses from matrix.

Conclusion: Similar to calcium alginate pellets, alginate-chitosan pellets demonstrated sustained drug release property though via different mechanisms. Combination of alginate, chitosan and calcium acetate in the same matrix nevertheless failed to retard drug release via complementary drug release pattern.     

Spray-spinning: a novel method for making alginate/chitosan fibrous scaffold

The subject of our investigations was the process of obtaining alginate/chitosan polyelectrolyte complex (PEC) fibers. In this study, a novel method named “spray-spinning” was developed for the making of these hybrid fibers. In spray-spinning, a chitosan solution was sprayed into a flowing sodium alginate solution and sheared into streamlines. The elongated streamlines subsequently transformed into alginate/chitosan PEC fibers. Average diameter of the fibers increased with the increasing of chitosan concentration used in spinning. The fibers showed a high water-absorbability of about 45 folds of water to their dry weight and retained their integrity after incubation in Minimum Essential Medium (MEM) for up to 30 days. In vitro co-culture experiments indicated that the fibers could support the three-dimensional growth of HepG2 cells and did not display any cyto-toxicity. Moreover, in vivo implanting experiments indicated that the connective tissue cells infiltrated into the implanted fibrous scaffolds in 3 weeks after surgery. These results demonstrated the potential applications of the as-spun fibers in regenerative medicine and tissue engineering.     

In vitro and in vivo evaluation of chitosan–gelatin scaffolds for cartilage tissue engineering

Chitosan–gelatin polyelectrolyte complexes were fabricated and evaluated as tissue engineering scaffolds for cartilage regeneration in vitro and in vivo. The crosslinker for the gelatin component was selected among glutaraldehyde, bisepoxy, and a water-soluble carbodiimide (WSC) based upon the proliferation of chondrocytes on the crosslinked gelatin. WSC was found to be the most suitable crosslinker. Complex scaffolds made from chitosan and gelatin with a component ratio equal to one possessed the proper degradation rate and mechanical stability in vitro. Chondrocytes were able to proliferate well and secrete abundant extracellular matrix in the chitosan–gelatin (1:1) complex scaffolds crosslinked by WSC (C1G1_WSC) compared to the non-crosslinked scaffolds. Implantation of chondrocytes-seeded scaffolds in the defects of rabbit articular cartilage confirmed that C1G1_WSC promoted the cartilage regeneration. The neotissue formed the histological feature of tide line and lacunae in 6.5 months. The amount of glycosaminoglycans in C1G1_WSC constructs (0.187 ± 0.095 μg/mg tissue) harvested from the animals after 6.5 months was 14 wt.% of that in normal cartilage (1.329 ± 0.660 μg/mg tissue). The average compressive modulus of regenerated tissue at 6.5 months was about 0.539 MPa, which approached to that of normal cartilage (0.735 MPa), while that in the blank control (3.881 MPa) was much higher and typical for fibrous tissue. Type II collagen expression in C1G1_WSC constructs was similarly intense as that in the normal hyaline cartilage. According to the above results, the use of C1G1_WSC scaffolds may enhance the cartilage regeneration in vitro and in vivo.     

Stability of alginate microbead properties in vitro

Alginate microbeads have been investigated clinically for a number of therapeutic interventions, including drug delivery for treatment of ischemic tissues, cell delivery for tissue regeneration, and islet encapsulation as a therapy for type I diabetes. The physical properties of the microbeads play an important role in regulating cell behavior, protein release, and biological response following implantation. In this research alginate microbeads were synthesized, varying composition (mannuronic acid to guluronic acid ratio), concentration of alginate and needle gauge size. Following synthesis, the size, volume fraction, and morphometry of the beads were quantified. In addition, these properties were monitored over time in vitro in the presence of varying calcium levels in the microenvironment. The initial volume available for solute diffusion increased with alginate concentration and mannuronic (M) acid content, and bead diameter decreased with M content but increased with needle diameter. Interestingly, microbeads eroded completely in saline in less than 3 weeks regardless of synthesis conditions much faster than what has been observed in vivo. However, microbead stability was increased by the addition of calcium in the culture medium. Beads synthesized with low alginate concentration and high G content exhibited a more rapid change in physical properties even in the presence of calcium. These data suggest that temporal variations in the physical characteristics of alginate microbeads can occur in vitro depending on synthesis conditions and microbead environment. The results presented here will assist in optimizing the design of the materials for clinical application in drug delivery and cell therapy.     

聚氨基酸复合微胶囊对Hb的控制释放

以生物相容性好、价格低廉的海藻酸钠(ALG)为聚阴离子芯材,通过静电液滴装置制备了平均粒径在290 μm左右、球形度好、表面光洁的海藻酸钙胶珠;再将生物可降解、具有介入治疗作用的聚精氨酸(PLA)与聚组氨酸(PLH)的混合物作为聚阳离子壁材,在海藻酸钙胶珠表面覆上一层高分子聚合膜以制备聚氨基酸复合微胶囊;并以牛血红蛋白Hb为药物模型,对微胶囊的控制释放性能进行了考察并将其初步应用于体外模拟口服给药。结果表明:聚氨基酸复合微胶囊在前0.5 h的累积释放量均低于40％,溶出结束时累积释放量均达到80%以上;ALG/(PLA-PLH)复合微胶囊和ALG/PLH微胶囊的药物释放速率均低于ALG/PLA微胶囊;于10 min成膜时间内制备的微胶囊具有较高的载药量、包封率和缓释性能;以pH 4.6 HAc-NaAc缓冲液为成膜溶媒制备的微胶囊,Hb持续释放时间和残留量均高于蒸馏水组;前2 h在模拟胃液的pH 1.2 HCl溶媒中累计释放的Hb不超过10％且绝大部分是在模拟肠液环境即pH 6.8 PBS 溶媒中释放的;壳聚糖的引入能在一定程度上延长药物释放时间。聚氨基酸复合微胶囊具备一定的缓释性、pH响应性和生理黏附性,有望成为一种口服给药系统用药物载体。      

In vitro study of the SBF and osteoblast-like cells on hydroxyapatite/chitosan–silica nanocomposite

Hydroxyapatite/chitosan–silica (HApCSi) nanocomposites were synthesized by co-precipitated method and their potential application as filler materials for bone regeneration were investigated in simulated body fluid (SBF). To study their biocompatibility, they were cultured with rat osteoblast-like UMR-106 cells for 3, 7, 14, and 21 days. Studies of the silica contents in chitosan matrix showed the presence of silinol (Si–OH) groups in CSi hybrid and their decrease after being composited with calcium phosphate (CaP) which is desirable for the formation of the apatite. XRD and TEM studies showed that the HAp formed in the CSi matrix were nanometer (20–40 nm) in size. Nanocomposites of HApCSi20 processed with 20%v/v silica whisker showed a micro hardness of 84.7 ± 3.3 MPa. Mineralization study in SBF showed the formation of apatite crystals on the HApCSi surface after being incubated for 7 days. In vitro biocompatibility, cell morphology, proliferation, and cell adhesion tests confirmed the osteoblast attachment and growth on the HApCSi20 surface. The density of cells and the production of calcium nodules on the substrate were seen to increase with increasing cultured time. The mechanical evaluation and in vitro experiment suggested that the use of HApCSi composite will lead to the formation of new apatite particles and thus be a potential implant material.     

Evaluation of alginate-chitosan semi IPNs as cartilage scaffolds

In this study, alginate and alginate:chitosan semi interpenetrating polymer network (IPN) scaffolds were prepared by freeze-drying process. Alginate scaffolds were crosslinked with different concentrations of CaCl₂, i.e. 0.5, 1 or 3% (w/v), in 96% (v/v) ethanol solutions for two different periods, i.e. 4 and 24 h, after freeze-drying. Scanning electron microscope (SEM)/ Energy Dispersive Analysis by X-ray (EDAX) analysis and swelling studies indicated that crosslinking of scaffolds with 3% (w/v) CaCl₂ for 24 h was effectively created suitable alginate scaffolds in terms of optimum porosity and mechanical stability. This is why, alginate:chitosan semi IPN scaffolds were prepared at the crosslinking condition mentioned above in 70:30, 60:40 and 50:50% (v/v) alginate:chitosan ratios. Besides the attachment and proliferation abilities of ATDC5 murine chondrogenic cells on alginate, 70:30% (v/v) alginate:chitosan and 50:50% (v/v) alginate:chitosan scaffolds, their cellular responses were assessed for chondrogenic potential. These structural and cellular outcomes demonstrate potential utility of chitosan semi IPNs in alginate scaffolds. Comparative results found in relation to alginate scaffolds, support the necessity for alginate:chitosan scaffolds for improved cartilage tissue engineering.     

Silk‐based microcarriers: current developments and future perspectives

Cell‐seeded microcarriers (MCs) are currently one of the most promising topics in biotechnology. These systems are supportive structures for cell growth and expansion that allow efficient nutrient and gas transfer between the media and the attached cells. Silk proteins have been increasingly used for this purpose in the past few years due to their biocompatibility, biodegradability and non‐toxicity. To date, several silk fibroin spherical MCs in combination with alginate, gelatin and calcium phosphates have been reported with very interesting outcomes. In addition, other silk‐based three‐dimensional structures such as microparticles with chitosan and collagen, as well as organoids, have been increasingly studied. In this study, the physicochemical and biological properties of these biomaterials, as well as the recent methodologies for their processing and for cell culture, are discussed. The potential biomedical applications are also addressed. In addition, an analysis of the future perspectives is presented, where the potential of innovative silk‐based MCs processing technologies is highlighted.Inspec keywords: biodegradable materials, proteins, calcium compounds, gelatin, biomedical materials, cellular biophysics, molecular biophysicsOther keywords: supportive structures, cell growth, gas transfer, attached cells, silk proteins, biodegradability, nontoxicity, silk fibroin spherical MCs, gelatin, calcium phosphates, silk‐based three‐dimensional structures, chitosan, collagen, physicochemical properties, biological properties, cell culture, silk‐based microcarriers, cell‐seeded microcarriers, biotechnology, efficient nutrient transfer, biocompatibility, alginate, biomedical applications     

Differential physical, rheological, and biological properties of rapid in situ gelable hydrogels composed of oxidized alginate and gelatin derived from marine or porcine sources

Marine derived gelatin is not known to associate with any communicable diseases to mammals and could be a reasonable substitute for gelatin derived from either bovine or porcine sources. The low melting point of marine gelatin (8°C) also offers greater formulation flexibility than mammalian derived gelatins. However, the sub-optimal physical properties of marine gelatin generally limit the interest to further develop it for biomedical applications. This study aimed at investigating the feasibility of using oxidized alginate (Oalg) as a high activity macromolecular crosslinker of marine gelatin to formulate in situ gelable hydrogels with the goal of enhancing the latter’s physical properties. The performance of Oalg/marine gelatin hydrogel was compared to Oalg/porcine gelatin hydrogel; in general, the physicomechanical properties of both hydrogels were comparable, with the hydrogels containing porcine gelatin exhibiting moderately higher mechanical strengths with shorter gelation times, smaller size pores, and higher swelling ratios. On the contrary, the biological performances of the two hydrogels were significantly difference. Cells cultured in the marine gelatin derived hydrogel grew significantly faster, with greater than 60% more cells by 7 days and they exhibited more spread-out conformations as compared those cultured in the porcine derived hydrogel. Production of ECM by cells cultured in the Oalg/marine gelatin hydrogel was up to 2.4 times greater than that of in the Oalg/porcine gelatin hydrogel. The biodegradation rate of the hydrogel formulated from marine gelatin was greater than its counterpart prepared from porcine gelatin. These differences have important implications in the biomedical applications of the two hydrogels. Huijuan Liao and Hanwei Zhang are the first authors.