الفهرس 
IntroductionOnly 14 pages are availabe for public view
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Abstract
Aim Of the Work is to:
• Establishment of the best formula which provides the total caloric requirement of critically ill neonate, with ɷ6: ɷ3 ratios lower than conventional used one (SMOF lipid®) and least complications on long term usage.
1- Study the formulation, characterizations, and stability of the selected formulae.
2- To evaluate the physicochemical stability of TPN mixtures, successfully applied in treatment of newborns and young children,
3- To analyses the possible interactions between the components,
4- To compare the kinetic and chemical stability of TPN admixtures containing two kinds of triglycerides (medium and long-chain triglycerides).
5- To monitor the stability of TPNs as a function of storage conditions
(Temperature, storage time).
The USP defines emulsions as follows: “Emulsions are two-phase systems in which one liquid is dispersed throughout another liquid in the form of small droplets. where oil is the dispersed phase and an aqueous solution is the continuous phase, the system is designated as an oil-in-water (O/W) emulsion. Conversely, where water or an aqueous solution is the dispersed phase and oil or oleaginous material is the continuous phase, the system is designated as a water-in-oil (W/O) emulsion”. More complicated emulsions such as O/W/O (i.e., oil droplets contained within aqueous droplets dispersed in a continuous oil phase) or W/O/W are also possible.
The type of emulsion is determined mainly by the volume ratio of the two liquids, the order of addition and the nature of the emulsifying agent.
Most pharmaceutical emulsions designed for oral administration are of the O/W type, emulsified lotions and creams for topical applications are either O/W or W/O, depending on their use. For the parenteral administration, both W/O and O/W emulsions are applicable, but only the O/W type could be injected intravascularly.
Oils that are most often included in the composition of nanoemulsions for parenteral nutrition are soybean oil, olive oil, fish oil, coconut oil, and rarely sunflower oil or cottonseed oil. Since they are used for the purpose of parenteral nutrition, they must be highly purified and exempted from peroxides, pigments, and unsaponifiable matters such as sterols and polymers. The soybean and fish oil, olive oil, medium-chain triglycerides (MCTs) as well as Linseed oil will be further discussed in more details since they were used in this research.
Our hypothesis was that O/W NEs could do alteration that was impacted mainly by the droplet size of the NEs.
To test the hypothesis, the overall objective of the study was to develop O/W NEs with significantly different droplet size and similar droplet surface structure, and to compare their in vivo bio distributions. The specific aims of the study were as follows:
1. To prepare O/W NEs which have distinct ” ” " ~ " ” ”250 nm for the medium-size NE (ME) with narrow size distribution.
2. To evaluate the prepared NEs stability for their suitability for the in vivo study.
3. To identify an agent to label the NEs.
4. To validate the assay method for the labeling agent.
6. To establish the in vivo biological sample preparation and assay protocol.
7. To study and compare the bio-distribution of labeled NEs with distinct droplet sizes after intravenous administration.
Chapter I: Clinical Survey
A three-month prospective study on Minia University Pediatric hospital NICU for critically ill patients undergoing major surgeries, from January 2019 to April 2020 and two groups selected, as a control group A of (twenty cases with neonates undergoing major surgeries and didn’t require a TPN plans for more than 14 days) was compared with group B (twenty cases with neonates undergoing major surgeries required a TPN plans for more than 14 days).
Chapter II: preliminary nanoemulsion formulation (Statistical factorial Design)
This design enables testing of the main effects due to formulation factors of the emulsion, determination of the possibility of interactions within the model and the significance of these factors and the interaction terms. The factors were the emulsion components such as Lipoid E80, Oil mixtures, Lipoid Sodium oleate®, Pl-188 and Millipore water. The dependent variables were z-average, z-potential, polydispersity index, dynamic viscosity, and kinematic viscosity of the emulsions.
Based on the optimization results, the composition of the nanoemulsion with the smallest mean droplet diameter and the lowest increase of that diameter within 60 days was defined. The optimal nanoemulsion was defined by the set of formulation parameters including oil 20%, egg phospholipids 1.2%, Pl-188 0.6%, Sodium oleate 0.03%, Glycerol 2.5%, α-tocopherols 0.01%, Thioglycolic acid 0.01%, water for injections up to 100%, while the process parameters were five cycles of homogenization and the pressure of 500 bar.
from the factorial experimental design, we concluded that by comparing the obtained results, it can be concluded that the values obtained by characterizing the TPN admixture containing the optimal nanoemulsion F3, F8, F9 and F10 has the best results on stability of structured lipid emulsions of Linseed oil
Chapter III: Optimization of nanoemulsions formulation.
All the operations used in the preparation of the finished product represent standard techniques to produce intravenous nanoemulsions. The process is the current state. of-the-art in the production of nanoemulsions for parenteral nutrition.
Thirty combinations were prepared with the ratios of Oil: Surfactant as 4:1, 8:1, and 16:1. noticed that best ratio giving the most stable emulsion, according to the experimental design, was 16:1. The Surfactant/co-surfactant (0.6 mg) ratios were 2:1. These mixtures were then mixed with the oil phase to form an isotropic produced by high speed combined with high pressure homogenization method. Thus, each combination had a total of 10 samples with different proportions of oil, surfactants and co-surfactants.
- The oil phase was composed of the following components:
Soybean oil (LPS Oil 700) (SO), Fish oil, (FO), medium chain triglycerides (MCT), Migylol 812 N, olive oil and linseed oil all filtered through a 0.22 µm filter membrane. Then we add, Egg phospholipids with 80% phosphatidylcholine, Lipoid ® E80 – (EP) and antioxidant (α-tocopherol. Toc), and the second antioxidant thioglycolic acid (TA).
- The water phase was composed of the following components:
Hydro soluble Kolliphor® Pl-188, emulsifier (PI.188) as a second surfactant, then glycerol G, Sodium Oleate as co-surfactant (SOI), sodium hydroxide for pH adjustment 7 . 8.5 and Water used in the experiment was double distilled at 100%, w/w.
Chapter IV: Biological Study.
The four chosen formulae (F3, F8, F9 &F10) and a SMOF control group were injected into 5 groups, each group contains 10 rats.
Histo-pathological Evaluation Liver samples were fixed by 10% buffered formaldehyde and embedded in paraffin. They were stained using standard H&E staining techniques and then graded by a single non-blinded pathologist for steatosis and inflammation (acute and chronic) using the histological grading system of Li et al. The grades of steatosis were scored as follows: 0, no steatosis; 1, mild steatosis in < 33% of the liver; 2, moderate steatosis in 34–66% of the liver; 3, severe steatosis in > 66% of the liver. The grades of inflammatory cell infiltration were the following: 0, none; 1, less than 5 foci/field; 2, 5 or more foci/field. Serum Biochemical Analysis The serum was prepared by centrifugation (10 min, 3000 rpm) and was stored at − 80 °C until the tests were analyzed. Serum total and direct bilirubin, aspartate transaminase (AST), and alanine transaminase (ALT) were determined with an automated chemistry analyzer (Olympus AU400, Olympus, Tokyo, Japan). Procedures were scaled up or down based on the manufacturer’s instructions.
Conclusion and recommendations
from all previously discussed data and experiment we concludes that the SLL (structure-Lipid of Linseed oil) is s lab oil mixture with best ɷ 6 : ɷ 3 ratio prepared by nanoemulsion technique, and after all characterizations and stability studies, may be effective for long term use of critically ill neonates undergoing abdominal surgeries and need a nutritional support for more than 14 days according to Experiments on rats and F3 formula may have the best composition that ensure the long last stability and efficacy, meanwhile, a clinical human studies and many further examinations are required.