Low-temperature 3D-printed collagen/chitosan scaffolds loaded with exosomes derived from neural stem cells pretreated with insulin growth factor-1 enhance neural regeneration after traumatic brain injury

There are various clinical treatments for traumatic brain injury,including surgery,drug therapy,and rehabilitation therapy;howeve r,the therapeutic effects are limited.Scaffolds combined with exosomes represent a promising but challenging method for improving the repair of traumatic brain injury.In this study,we determined the ability of a novel 3D-printed collagen/chitosan scaffold loaded with exosomes derived from neural stem cells pretreated with insulin-like growth factor-1(3D-CC-INEXOS) to improve traumatic brain injury repair and functional recove ry after traumatic brain injury in rats.Composite scaffolds comprising collagen,chitosan,and exosomes derived from neural stem cells pretreated with insulin-like growth fa ctor-1(INEXOS) continuously released exosomes for 2weeks.Transplantation of 3D-CC-INExos scaffolds significantly improved motor and cognitive functions in a rat traumatic brain injury model,as assessed by the Morris water maze test and modified neurological seve rity scores.In addition,immunofluorescence staining and transmission electron microscopy showed that3D-CC-INExos implantation significantly improved the recove ry of damaged nerve tissue in the injured area.In conclusion,this study suggests that transplanted3D-CC-INExos scaffolds might provide a potential strategy for the treatment of traumatic brain injury and lay a solid foundation for clinical translation.

Chitosan conduits enriched with fibrin-collagen hydrogel with or without adipose-derived mesenchymal stem cells for the repair of 15-mm-long sciatic nerve defect

Hollow conduits of natural or synthetic origins have shown acceptable regeneration results in short nerve gap repair;however,results are still not comparable with the current gold standard technique“autografts”.Hollow conduits do not provide a successful regeneration outcome when it comes to critical nerve gap repair.Enriching the lumen of conduits with different extracellular materials and cells could provide a better biomimicry of the natural nerve regenerating environment and is expected to ameliorate the conduit performance.In this study,we evaluated nerve regeneration in vivo using hollow chitosan conduits or conduits enriched with fibrin-collagen hydrogels alone or with the further addition of adipose-derived mesenchymal stem cells in a 15 mm rat sciatic nerve transection model.Unexpected changes in the hydrogel consistency and structural stability in vivo led to a failure of nerve regeneration after 15 weeks.Nevertheless,the molecular assessment in the early regeneration phase(7,14,and 28 days)has shown an upregulation of useful regenerative genes in hydrogel enriched conduits compared with the hollow ones.Hydrogels composed of fibrin-collagen were able to upregulate the expression of soluble NRG1,a growth factor that plays an important role in Schwann cell transdifferentiation.The further enrichment with adipose-derived mesenchymal stem cells has led to the upregulation of other important genes such as ErbB2,VEGF-A,BDNF,c-Jun,and ATF3.

Comparative study of chitosan/fibroin–hydroxyapatite and collagen membranes for guided bone regeneration in rat calvarial defects: micro-computed tomography analysis

This study aimed to utilize micro-computed tomography （micro-CT） analysis to compare new bone formation in rat calvarial defects using chitosan/fibroin-hydroxyapatite （CFB-HAP） or collagen （Bio-Gide） membranes. Fifty-four （54） rats were studied. A circular bony defect （8 mm diameter） was formed in the centre of the calvaria using a trephine bur. The CFB-HAP membrane was prepared by thermally induced phase separation. In the experimental group （n= 18）, the CFB-HAP membrane was used to cover the bony defect, and in the control group （n= 18）, a resorbable collagen membrane （Bio-Gide） was used. In the negative control group （n= 18）, no membrane was used. In each group, six animals were euthanized at 2, 4 and 8 weeks after surgery. The specimens were then analysed using micro-CT. There were significant differences in bone volume （BV） and bone mineral density （BMD） （P〈O.05） between the negative control group and the membrane groups. However, there were no significant differences between the CFB-HAP group and the collagen group. We concluded that the CFB-HAP membrane has significant potential as a guided bone regeneration （GBR） membrane.

Collagen-chitosan scaffold impregnated with bone marrow mesenchymal stem cells for treatment of traumatic brain injury

Combinations of biomaterials and cells can effectively target delivery of cells or other therapeutic factors to the brain to rebuild damaged nerve pathways after brain injury.Porous collagen-chitosan scaffolds were prepared by a freeze-drying method based on brain tissue engineering.The scaffolds were impregnated with rat bone marrow mesenchymal stem cells.A traumatic brain injury rat model was established using the 300 g weight free fall impact method.Bone marrow mesenchymal stem cells/collagen-chitosan scaffolds were implanted into the injured brain.Modified neurological severity scores were used to assess the recovery of neurological function.The Morris water maze was employed to determine spatial learning and memory abilities.Hematoxylin-eosin staining was performed to measure pathological changes in brain tissue.Immunohistochemistry was performed for vascular endothelial growth factor and for 5-bromo-2-deoxyuridine(BrdU)/neuron specific enolase and BrdU/glial fibrillary acidic protein.Our results demonstrated that the transplantation of bone marrow mesenchymal stem cells and collagen-chitosan scaffolds to traumatic brain injury rats remarkably reduced modified neurological severity scores,shortened the average latency of the Morris water maze,increased the number of platform crossings,diminished the degeneration of damaged brain tissue,and increased the positive reaction of vascular endothelial growth factor in the transplantation and surrounding areas.At 14 days after transplantation,increased BrdU/glial fibrillary acidic protein expression and decreased BrdU/neuron specific enolase expression were observed in bone marrow mesenchymal stem cells in the injured area.The therapeutic effect of bone marrow mesenchymal stem cells and collagen-chitosan scaffolds was superior to stereotactic injection of bone marrow mesenchymal stem cells alone.To test the biocompatibility and immunogenicity of bone marrow mesenchymal stem cells and collagen-chitosan scaffolds,immunosuppressive cyclosporine was intravenously injected 12 hours before transplantation and 1-5 days after transplantation.The above indicators were similar to those of rats treated with bone marrow mesenchymal stem cells and collagen-chitosan scaffolds only.These findings indicate that transplantation of bone marrow mesenchymal stem cells in a collagen-chitosan scaffold can promote the recovery of neuropathological injury in rats with traumatic brain injury.This approach has the potential to be developed as a treatment for traumatic brain injury in humans.All experimental procedures were approved by the Institutional Animal Investigation Committee of Capital Medical University,China(approval No.AEEI-2015-035)in December 2015.

Effect of chitosan/type I collagen/gelatin composites in biocompatibility and nerve repair

Chitosan, collagen I and gelatin were mixed in appropriate quantities to develop a new nerve repair material, with good arrangement and structure, as well as even aperture size. The composite material was sterilized by 60Co irradiation for 24 hours prior to implantation in the right thigh of rats following sciatic nerve damage. Results showed that the material was nontoxic to the kidneys and the liver, and did not induce an inflammatory response in the muscles. The composite material enhanced the recovery of sciatic nerve damage in rats. These experimental findings indicate that the composite material offers good biocompatibility and has a positive effect on injured nerve rehabilitation.

Preparation of a Cyclomaltoheptaose(β-cyclodextrin) Cross-linked Chitosan Derivative via Glyoxal or Glutaraldehyde

A cyclomaltoheptaose--cyclodextrin (-CD) crosslinked chitosan derivative via glyoxal or glutaraldehyde was prepared. The structures of -CD crosslined chitosan with glyoxal or glutaraldehyde were characterized by IR spectra. The surface morphology of the -CD crosslinked chitosan particles was examined using a scanning electron microscope. The immobilization capacity of ?CD on chitosan was affected on the weight ratio of -CD/chitosan, the utilization amount of crosslinking agent, the acidity of the reaction system and the temperature. The adsorption for nicotine indicated that the chitosan--CD was a good adsorbent.

Collagen/Chitosan Electrospun Nanofibrous Membrane for Wound Dressing

Collagen(Col)/chitosan(CS)nanofibrous membrane has great potential to be used as wound dressing.However,current Col/CS nanofibrous membrane produced from electrospinning can not offer sufficient mechanical strength for practical applications.Herein,a novel mixed solvent was used to prepare next-generation high-strength Col/CS nanofibrous membrane.Meanwhile,the optimal Col to CS weight ratio was investigated as well.The asproduced membrane was examined by scanning electron microscopy(SEM),attenuated total reflectance Fourier transform infrared spectroscopy(ATR-FTIR),differential scanning calorimetry(DSC),and XF-1A tester to study its morphological,chemical,thermal and mechanical properties.The preliminary results demonstrated that the mechanical properties of Col/CS nanofibrous membranes were enhanced substantially with the increase of CS weight ratios from 0 to 90%and the optimal Col to CS weight ratio was determined to be 1∶1.A promising way was presented to fabricate Col/CS electrospun nanofibrous membrane with sufficient mechanical strength for practical wound dressing applications.

Preparation and characterization of β-cyclodextrin grafted N-maleoyl chitosan nanoparticles for drug delivery

β-cyclodextrin (CD) grafted N-maleoyl chitosan (CD-g-NMCS) with two different degrees of substitution (DS) of N-maleoyl (DS = 21.2% and 30.5%) were synthesized from maleic anhydride and chitosan bearing pendant cyclodextrin (CD-g-CS). CD-g-NMCS based nanoparticles were prepared via an ionic gelation method together with chitosan and CD-g-CS nanoparticles.The size and zeta potential of prepared CD-g-NMCS nanoparticles were 179.2~274.0 nm and 36.2~42.4 m V, respectively. In vitro stability test indicated that CD-g-NMCS nanoparticles were more stable in phosphate-buffered saline compared with chitosan nanoparticles. Moreover, a poorly water-soluble drug, ketoprofen (KTP), was selected as a model drug to study the obtained nanoparticle’s potentials as drug delivery carriers. The drug loading efficiency of CD-g-NMCS20 nanoparticles were 14.8% for KTP. MTT assay showed that KTP loaded CD-g-NMCS nanoparticles were safe drug carriers. Notably, in vitro drug release studies showed that KTP was released in a sustained-release manner for the nanoparticles. The pharmacokinetic of drug loaded CD-g-NMCS20 nanoparticles were evaluated in rats after intravenous administration. The results of studies revealed that, compared with free KTP, KTP loaded CD-g-NMCS20 nanoparticles exhibited a significant increase in AUC0→24h and mean residence time by 6.6-fold and 2.9-fold, respectively. Therefore, CD-g-NMCS nanoparticles could be used as a novel promising nanoparticle-based drug delivery system for sustained release of poorly water-soluble drugs. The carboxylic acid groups of the CD-g-NMCS molecule provide convenient sites for further structural modifications including introduction of tissue-or disease-specific targeting groups.

Synthesis of Novel Chitosan Microspheres Grafted with β-Cyclodextrins and Their Adsorption for Iodine for Antibacterial Activities

The novel chitosan microspheres grafted with beta-cyclodextrins (CMGC) wereprepared by means of the reaction of chitosan microspheres and mono-(6-p-tosyl)- beta-cyclodextrins(beta-CD-OTs-6). beta-CD-OTs-6 were gained by the reaction of p-toluenesul-fonyl chloride (TsCl) andbeta-cyclodextrins (beta-CDs). Their structures were proved by Fourier transform infrared spectralanalysis (FT-IR), X ray powder diffraction analysis, and ^(13)C NMR; the configuration of CMGC wascharacterized by scanning electron micrograph (SEM) and transmittance electron micrograph (TEM). Theinclusion complex of CMGC with iodine was prepared and its inclusion ability was studied. Theexperimental results showed that some iodine was included with CMGC and formed a stable inclusion.The stable complex of CMGC and iodine (CMGC-1) shows good antibacterial effect.

Chitosan-collagen porous scaffold and bone marrow mesenchymal stem cell transplantation for ischemic stroke

In this study, we successfully constructed a composite of bone marrow mesenchymal stem cells and a chitosan-collagen scaffold in vitro, transplanted either the composite or bone marrow mesenchymal stem cells alone into the ischemic area in animal models, and compared their effects. At 14 days after co-transplantation of bone marrow mesenchymal stem cells and the hi- tosan-collagen scaffold, neurological function recovered noticeably. Vascular endothelial growth factor expression and nestin-labeled neural precursor cells were detected in the iscbemic area, surrounding tissue, hippocampal dentate gyrus and subventricular zone. Simultaneously, a high level of expression of glial fibrillary acidic protein and a low level of expression of neuron-spe- cific enolase were visible in BrdU-labeled bone marrow mesenchymal stem cells. These findings suggest that transplantation of a composite of bone marrow mesenchymal stem cells and a chi- tosan-collagen scaffold has a neuroprotective effect following ischemic stroke.

Preparation and Evaluation of Insulin-loaded Nanoparticles based on Hydroxypropyl-β-cyclodextrin Modified Carboxymethyl Chitosan for Oral Delivery

Novel insulin-loaded nanoparticles based on hydroxypropyl-β-cyclodextrin modified carboxymethyl chitosan（CMC-HP-β-CD） were prepared to improve the oral bioavailability of insulin. The CMC-HP-β-CD was characterized by FT-IR spectroscopy and 1H-NMR spectra. The insulin-loaded nanoparticles were prepared through crosslinking with calcium ions, and the morphology and size of the prepared nanoparticles were characterized by transmission electron microscopy（TEM） and dynamic light scattering（DLS）. Cumulative release in vitro study was performed respectively in simulated gastric medium fluid（SGF, p H=1.2）, simulated intestinal fluid（SIF, p H=6.8） and simulated colonic fluid（SCF, p H=7.4）. The encapsulation efficiency of insulin was up to 87.14 ± 4.32% through high-performance liquid chromatography（HPLC）. Statistics indicated that only 15% of the encapsulated insulin was released from the CMC-HP-β-CD nanoparticles in 36 h in SGF, and about 50% of the insulin could be released from the nanoparticles in SIF, whereas more than 80% was released in SCF. In addition, the solution containing insulin nanoparticles could effectively reduce the blood glucose level of diabetic mice. The cytotoxicity test showed that the samples had no cytotoxicity. CMC-HP-β-CD nanoparticles are promising candidates as potential carriers in oral insulin delivery systems.

Preparation,Characterization of Hydrophobic Drug in Combine Loaded Chitosan/Cyclodextrin/Trisodium Citrate Nanoparticles and in vitro Release Study

Chitosan/cyclodextrin/trisodium citrate（CS/CD/TSC） nanoparticles with ibuprofen（IBU） loaded were prepared via the ionic cross-linking method, with trisodium citrate selected as the cross-linking agent. The drug-loading capacity, particle size, zeta potential and surface morphology of the obtained nanoparticles were inves tigated. The results show a good drug-loading capacity. The prepared nanoparticles were spherical in shape with an average size of 293.7 nm and a zeta potential of ＋30.72 mV. The in vitro release studies show that the controlled re lease of IBU from the nanoparticles was followed. The drug release from CS/β-CD/TSC nanoparticles followed non-Fickian or anomalous diffusion.

Mechanical Properties of Collagen Biomimetic Films Formed in the Presence of Calcium,Silica and Chitosan

Using eucollagen solutions from ox hide, we cast collagen films to assess the influence of calcium and silica on the reconstitution of the fibrous structure of collagen. The tensile strength and the breaking elongation of the reconstituted collagen films were measured and analysed. Significant differences were observed between reconstituted collagen films with and without calcium and silica. The breaking elongation of the films obtained in the presence of silica was significantly greater, and the degradation was lower than other films of reconstituted collagen. Collagen and chitosan do not exist together as blends in nature, but the specific properties of each may be used to produce in biomimetic way man-made blends with biomedical applications, that confer unique structural, mechanical （detail） and in vivo properties.

Electrospun Nanofibers of Hydroxyapatite / Collagen / Chitosan Promote Osteogenic Differentiation of BMSCs

Bone tissue engineering, aiming at developing bone substitutes for repair and regeneration of bone defects instead of using autologous bone grafts,has attracted wide attention in the field of tissue engineering and regenerative medicine.Developing biomimetic biomaterial scaffolds able to regulate osteogenic differentiation of stem cells could be a promising strategy to improve the therapeutic efficacy.In this study, electrospun composite nanofibers of hydroxyapatite / collagen / chitosan( HAp / Col / CTS)resembling the fibrous nanostructure and constituents of the hierarchically organized natural bone,were prepared to investigate their capacity for promoting bone mesenchymal stem cells( BMSCs)to differentiate into the osteogenic lineage in the absence and presence of the osteogenic supplementation, respectively.Cell morphology,proliferation and quantified specific osteogenic protein expression on the electrospun HAp / Col / CTS scaffolds were evaluated in comparison with different controls including electrospun nanofibrous CTS,HAp / CTS and tissue culture plate.Our results showed that the nanofibrous HAp / Col / CTS scaffolds supported better spreading and proliferation of the BMSCs than other substrates( P < 0.01).Expressions of osteogenesis protein markers,alkaline phosphatase( ALP) and Col,were significantly upregulated on the HAp / Col / CTS than those on the CTS( P < 0.01) and HAp /CTS( P < 0.05) scaffolds in the absence of the osteogenic supplementation.Moreover,presence of osteogenic supplementation also proved to enhance osteogenic differentiation of BMSCs on HAp /Col / CTS scaffolds, indicative of a synergistic effect.This study highlights the potential of BMSCs / HAp / Col / CTS cell-scaffold system for functional bone repair and regeneration applications.

Elaboration of Materials with Functionality Gradients by Assembly of Chitosan-Collagen Microspheres Produced by Microfluidics

Biopolymers extracted from renewable resources like chitosan and collagen exhibit interesting properties for the elaboration of materials designed for tissue engineering applications,among which are their hydrophilicity,biocompatibility and biodegradability.In many cases,functional recovery of an injured tissue or organ requires oriented cell outgrowth,which is particularly critical for nerve regeneration.Therefore,there is a growing interest for the elaboration of materials exhibiting functionalization gradients able to guide cells.Here,we explore an original way of elaborating such gradients by assembling particles from a library of functionalized microspheres.We propose a simple process to prepare chitosan-collagen hybrid microspheres by micro-and milli-fluidics,with adaptable dimensions and narrow size distributions.The adhesion and survival rate of PC12 cells on hybrid microspheres were compared to those on pure chitosan ones.Finally,functionalized microspheres were assembled into membranes exhibiting a functionalization gradient.

Biological Feature of Collagen-Chitosan Membrane with Basic Fibroblast Growth Factor for the Culture of Human Fibroblast

The effect of collagen-chitosan membrane with different proportionate collagen and bFGF were investigated for culture human fibroblast. The optimum weight ratio of collagen/chitoson and bFGF were selected. Using culture human fibroblast technologies and cytotoxicity evaluated in vitro, Cell morphology was observed. Experimental results show that collagen-chitosan with bFGF promoted human fibroblast adhesion and supported cell proliferation for a long time. Furthermore collagen-chitosan membrane obviously degrade after 18d when human fibroblast was exhibited fusion spreading, compacting and stabilize. Cytotoxic to human fibroblast was revealed very low . Collagen- chitosan with bFGF should be useful as a tissue engineering biomaterial scaffold for cell culture.

Cyclodextrin/chitosan nanoparticles for oral ovalbumin delivery: Preparation, characterization and intestinal mucosal immunity in mice

A novel oral protein delivery system with enhanced intestinal penetration and improved antigen stability based on chitosan(CS) nanoparticles and antigen-cyclodextrin(CD) inclusion complex was prepared by a precipitation/coacervation method. Ovalbumin(OVA) as a model antigen was firstly encapsulated by cyclodextrin, either β-cyclodextrin( β-CD) or carboxymethyl-hydroxypropyl-β-cyclodextrin(CM-HP-β-CD) and formed OVA-CD inclusion complexes, which were then loaded to chitosan nanoparticles to form OVA loaded β-CD/CS or CM-HP-β-CD/CS nanoparticles with uniform particle size(836.3 and 779.2 nm, respectively) and improved OVA loading efficiency(27.6% and 20.4%, respectively). In vitro drug release studies mimicking oral delivery condition of OVA loaded CD/CS nanoparticles showed low initial releases at p H 1.2 for 2 h less than 3.0% and a delayed release which was below to 30% at p H 6.8 for further 72 h. More importantly, after oral administration of OVA loaded β-CD/CS nanoparticles to Balb/c mice, OVA-specific sIgA levels in jejunum of OVA loaded β-CD/CS nanoparticles were 3.6-fold and 1.9-fold higher than that of OVA solution and OVA loaded chitosan nanoparticles, respectively. In vivo evaluation results showed that OVA loaded CD/CS nanoparticles could enhance its efficacy for inducing intestinal mucosal immune response. In conclusion, our data suggested that CD/CS nanoparticles could serve as a promising antigen-delivery system for oral vaccination.

羟基磷灰石-聚乙烯醇/胶原-壳聚糖-明胶复合水凝胶修复兔骨软骨缺损

背景:关节骨软骨缺损是目前骨科医生面临的难题,传统修复方法难以取得满意疗效,以羟基磷灰石-聚乙烯醇为基础的复合水凝胶材料是目前研究的一个方向。目的:制备羟基磷灰石-聚乙烯醇/胶原-壳聚糖-明胶复合水凝胶,表征其物理学特征,验证其植入体内后的组织相容性及细胞黏附增殖能力,探讨其对兔骨软骨缺损的修复效果。方法:利用3D打印技术制备圆柱状多孔聚乳酸支架(孔径分别为1.2,1.4,1.6,1.8 mm),再将聚乙烯醇及羟基磷灰石混合乳液灌入聚乳酸支架中,经过冻融及二氯甲烷溶解后制成羟基磷灰石-聚乙烯醇水凝胶。然后将胶原-壳聚糖-明胶混合液灌入羟基磷灰石-聚乙烯醇水凝胶中,利用京尼平进行交联,最后经乙醇清洗及冷冻干燥制成羟基磷灰石-聚乙烯醇/胶原-壳聚糖-明胶复合水凝胶,表征4组水凝胶的物理学特征,筛选性能最优的水凝胶进行后续实验。将羟基磷灰石-聚乙烯醇水凝胶、胶原-壳聚糖-明胶复合水凝胶分别植入SD大鼠皮下,苏木精-伊红与Masson染色观察材料表面的细胞黏附生长状况。在15只兔双侧膝关节股骨滑车制作骨软骨缺损(直径5 mm、深6 mm)模型,左侧植入复合水凝胶(实验组),右侧未植入任何材料(对照组),Micro-CT及组织学检测来评估其对骨软骨缺损的修复效果。结果与结论:①综合孔隙率、含水率、力学测试及扫描电镜结果,得出孔径1.2 mm的羟基磷灰石-聚乙烯醇/胶原-壳聚糖-明胶复合水凝胶更符合天然软骨的一般特性,用于后续实验;②苏木精-伊红与Masson三色染色显示,随着材料植入大鼠皮下时间的延长,两种材料周边黏附的细胞均明显增多,其中胶原-壳聚糖-明胶植入后的细胞增殖状况更好,可见大量细胞长入其形成的网状结构,且网状结构也逐渐降解;③在兔骨软骨缺损实验中,至术后8周时,Micro-CT检查显示实验组植入的材料和周围骨-软骨整合良好,其表面及内部均有部分骨长入,对照组软骨及软骨下层仍然存在明显的缺损,未形成有效修复;苏木精-伊红与甲苯胺蓝染色显示,实验组植入复合水凝胶4-8周后即与周围骨软骨发生整合,随着时间的延长材料表面逐渐有新生软骨形成,至12周时缺损处大部分被新生软骨覆盖,软骨下层也出现良好的骨长入;对照组至术后12周虽然深层骨缺损大部分修复、浅层的软骨及软骨下骨缺损也有一定程度修复,但主要被纤维组织及部分纤维软骨替代;④结果表明,羟基磷灰石-聚乙烯醇/胶原-壳聚糖-明胶复合水凝胶材料可仿生天然软骨的结构和功能,在动物实验中能有效修复骨软骨缺损。

羧甲基壳聚糖对牛骨胶原蛋白微观结构、热稳定性及自组装性质的影响

构建具备良好热稳定性、自组装性质及生物相容性的可食性细胞外基质(extracellular matrix,ECM)类似物支架对于制造结构化细胞培养肉制品至关重要。将羧甲基壳聚糖(carboxymethyl chitosan,CMCS)引入牛骨胶原蛋白(bovine bone collagen,BBC)体系中,通过光谱分析(紫外、红外、荧光光谱)发现BBC与CMCS的相互作用随着引入CMCS添加量的增加而增强,但并未影响BBC的三螺旋结构。差示扫描量热法/热重分析结果表明,CMCS的引入增强了BBC体系的热稳定性。浊度试验及扫描电子显微镜/透射电子显微镜观察结果证实了CMCS引入后胶原蛋白纤维形成度呈上升趋势,聚集行为更明显且自组装速率产生变化,呈现出更疏松扭曲的三维结构以及更大的纤维直径及更广泛的直径分布。但CMCS的引入并未明显影响BBC的D-周期性结构(胶原纤维自组装过程中形成的特征性明暗交替的周期性横纹结构)形成及其长度,且CMCS引入前后体系的细胞相容性也未呈现显著性差异。随着引入CMCS添加量增加,CMCS和BBC之间的静电作用力可能较共价相互作用和氢键更占优势。这些结果表明,CMCS的引入不影响BBC三螺旋结构完整性和生物相容性,并改善了BBC的热稳定性及体外自组装性质。这为开发新型优良可食性胶原蛋白基ECM仿生支架在细胞培养肉领域的应用以及畜禽骨副产物高值化精深加工利用提供了参考信息。

胶原-壳聚糖微球的制备与载药性能研究

本文采用胶原与壳聚糖作为原材料,利用壳聚糖的pH响应性,制备了一种无添加交联剂的复合微球,并通过天狼星红染色法和羟脯氨酸分析法,测定复合微球中胶原的含量。此外,对微球的形貌、粒径、热稳定性以及持水性能进行了系统性分析。通过将姜黄素作为模拟药物,对微球的载药性能进行了探索。