| With the improvement of living standard,it has become a hot research hotspot in the field of nutrition and health food to obtain the functional bioactive components with physiological regulation to improve the human health.Therefore,it is very important to fabricate a suitable targeting controlled-release delivery system to improve the effectiveness and bioavailability of nutrients during processing and physiological conditions.Based on the special transport and immune response mechanism of M cell,the starch based self-assembled carrier material which is suitable for M-cell target delivery and its nanomicelles delivery system were developed through starch molecular hydrophobic and targeted modification.Dynamic changes of the interaction between starch based self-assembled carrier materials and immune functional nutrients under the condition of assembly process,storage and digestive tract and cell environment were controlled at the interaction level between the starch based self-assembled nanomicelles and intestinal M cells.Furthermore,the related regulation mechanism was revealed in this thesis.It has important academic value and guiding significance for improving the effectiveness of nutrition and function of healthy food.Based on the molecular interaction and self-assembly behavior and combined with the structural characteristics of nanomicelles and the specific receptors at the M-cell surface,the esterification reaction method is applied to regulate the substituted side chains of fatty acid esters and CC9 M cell targeting peptide and the assembly and aggregation behavior of starchbased carrier material molecules at the molecular level were induced.The effect of the structure characteristics and concentration of the starch based self-assembled carrier materials,assemble temperature,pH and ion strength on the self-assembly and aggregation behavior,stability of micellar particles,encapsulation of the immune polypeptide and M cell targeting and the relative mechanism were obtained,and the M-cell targeting amphiphilic starch self-assembly carrier materials were designed.The results showed that after the starch molecular modification by the hydrophobic side chain of the fatty acid ester,the increase of the length and the substituted degree of the hydrophobic side chain,the concentration of starch-based carriers,assembly temperature and ionic strength are favorable for the material assembly in aqueous solution and the negatively charged spherical micelle particles with an average particle size of 50~380 nm were formed.Compared with the particle size of self-assembled micelles formed by the laurate amphiphilic starch molecules,the particle size distribution of self-assembled micelles formed by the stearic ester amphiphilic starch molecules is relative uniform and the dilution stability and storage stability of micelle particles in aqueous solution were increased with the increase of the length and the degree of substitution of the hydrophobic side ester chains.The CC9 polypeptides with M-cell targeting which was selected by fluorescence labeling and laser confocal microscopy were introducted to the amphiphilic starch molecules and the formation of self-assembly micelles was systematic analyzed using modern analytical techniques.Effects of the material structure and concentration,pH,ionic strength,storage time and dilution on the self-assembly and aggregation behavior of M-cell targeting amphiphilic starch self-assembled materials in aqueous solution were obtained.The results showed that the particle size of the self-assembly micelle particles is relative uniform with the increase of the substituted CC9 targeting polypeptide.The particle size of the self-assembly micelle particles become narrow and larger micelle particles or micelle particle aggregates were formed with the increase of material concentration.The sensitivity of the M cell targeting hydrophilic starch self-assembled carrier materials to the pH,ionic strength,storage time and dilution was increased and the dilution stability of self-assembly micelle particles decreased but the storage stability increased compared with the fatty acid ester amphiphilic starch molecules.By controlling the degree of the grafted M-cell targeting polypeptide side chain,the pH and ionic strength,the ability of the M-cell target amphiphilic starch self-assembled carrier materials to form micelles in aqueous solution can be modulated and the M-cell target amphiphilic starch self-assembled carrier materials were selected for the self-assembled micelle particles with certain tolerance the changes of pH,ionic strength,dilution degree and suitable storage stability.The assembly between the M-cell target amphiphilic starch self-assembled carrier materials and the immune polypeptide molecules is regulated by controlling the hydrophobic interaction between the molecules.The intermolecular interactions,assembly and aggregation behavior between M-cell target amphiphilic-starch self-assembled carrier materials and polypeptide molecules and the M-cell targeting controlled release delivery characteristics and its regulation mechanism were revealed using the modern analysis techniques.Thus,a starchbased self-assembled nanomicelle delivery system with M-cell targeting is established.The results showed that SES-3 starch based self-assembled nanomicelle particle delivery system with good TRP2 polypeptide loading capability and stability was obtained by the control of the length and the substituted degree of the hydrophobic fatty acid esters,material concentration,temperature,pH and ionic strength for regulating the assembly behavior between the amphiphilic starch self-assembled carrier materials and TRP2 polypeptide.This nanomicelle particle delivery system can deliver about 59.25% TRP2 polypeptide to small intestinal epithelial cells.Furthermore,the changes of encapsulation of TRP2 polypeptide,self-assembled micelle particle size and distribution,intermolecular interaction between carrier material and TRP2 polypeptide,dilution stability and storage stability of micelle particles,the in-vitro releasing and M cell-targeting properties were systematically investigated.The results showed that the M-cell-targeted amphiphilic starch self-assembled carrier materials can formed a starch-based self-assembled nanomicelle particle delivery system with good TRP2 polypeptide loading capability,stability and M-cell targeted controlled release characteristics under pH 6.8 without NaCl and 0.1 mg/mL material concentration.This starch-based self-assembled nanomicelle particle delivery system can about 71.98% TRP2 polypeptide to small intestinal epithelial cells and significantly increase its target concentration and translocation at M cells.From the molecular level,the suitable M cell targeting amphiphilic starch self-assembly carrier was established by using the hydrophobic and M cell targeting modification to adjust its self-assembly behavior and the M cell targeting delivery behavior.Relationship among the structure of starch-based carriers and self-assembled nanomicelles and the M cell targeting delivery characteristics were established.The M cell targeting self-assembled nanomicelle delivery system with good loading capacity stability and M-cell targeted controlled release characteristics was obtained and its construction method were constructed.These results are expected to provide technical support and theoretical basis for M-cell targeting controlled release delivery of functional bioactive components and for enhancing their bioavailability. |
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