New advanced approach for processing natural polymers chitin and guar gum for developing new bioactive biocompatible drug delivery systems for oral administration of insulin

doi:10.21203/rs.3.rs-1765632/v1

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New advanced approach for processing natural polymers chitin and guar gum for developing new bioactive biocompatible drug delivery systems for oral administration of insulin

https://doi.org/10.21203/rs.3.rs-1765632/v1

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Two nanocomposites samples of oral administrations insulin-drug delivery systems: insulin@chitin and insulin@chitin-grafted (g)-guar gum are prepared by using a modified sol-gel method. The novelty of this preparations is that, it is the first time to load insulin vials on such safe ecofriendly natural polymers using water as green solvent. The traces number of chemical agents used in these preparations are below the toxicity limits. Native chitin and chitin-g-guar gum crosslinked copolymer are used as inert bioactive carriers for insulin vials. Guar gum increased the numbers of functional groups of the copolymer. Spectrophotometric monitoring of insulin release at physiological temperature, 37 ± 0.5ºC and buffer solutions pH 1.2, 6.8, 7.4. These tested aqueous solutions simulating biological body fluids of stomach, intestine colon and blood stream. Insulin release from insulin@chitin only at pH 7.4 as monitored by spectrophotometry. No insulin released can be monitored at pH 1.2, and 6.8 due strong bonding of insulin to acetyl group of chitin. In contrast, insulin@chitin- g-guar gum showed sustained targeting insulin-release followed the order: pH 6.8 > pH 7.4 > pH 1.2. The release data obeyed pseudo second order kinetic model indicating that this coplymer is heterogeneous solid surface of energetically different active sites and each insulin molecule occupied two active sites. Slow rate of insulin release at pH 1.2 indicated its protection against stomach juice. The kinetic results and the physicochemical characteristics porosity, swelling ratio, peptide&amino acid sequence, zeta potential, particle size showed that: insulin loaded on chitin by chemisorbed monolayer and it is loaded on the copolymer by chemi-, and phyi-sorption forces. Both insulin@chitin and insulin@chitin-g-GG nanocomposites showed negative zeta potential indicating adequate stability against coagulation. However, insulin@chitin-g-GG nanocomposite showed more monodispersed particle size distribution (than insulin@chitin) enhancing insulin release.

Chitin

guar gum

insulin

drug delivery system

release

rate

linear fitting

Chitin is smart natural polymer for formulations of drug delivery systems (DDS). It is available, biocompatible and biodegradable [1], an ecofriendly second abundant natural polysaccharide after cellulose obtained from renewable resources such as seaweeds, exoskeleton of anthoropods specially crustaceans like lobster, insects and internal shells of cephalopods such as squid and octopus. Its main source is wastes of crabs, and shrimp's shells or generally shellfish wastes.

Chitin is chemically extracted from crustacean shells via demineralization and deproteinization, supplementary information (SI), section I.1[2].The chemical structure of chitin is represented in, Fig.SI.1a, (β-(1→4 glycosidic bond)-N-acetyl-D-glucosamine), or 2-acetamido-2-deoxy-D-glucopyranose. The main functional groups are primary hydroxyl CH₂OH-, and acetamide CH₃CONH-groups. Chitin chains can form intramolecular hydrogen bonding.

Guar gum (GG) is: extracted from plant seeds; a linear poly- β-1,4 anhydro mannose -galactose residues, Fig.SI.1b. GG contains four hydroxyl groups per each monomeric unit [3]. It is extensively used in formulation of hydrogel biosorbents for wastewater treatment [4]. However, its uses in formulation of therapeutic DDS, especially for insulin is not reported yet in the literature.

Human insulin hormone, C₂₅₇H₃₈₃N₆₅O₇₇S₆ is a dipeptide protein synthesized, release and stored in pancreas. Dipeptide chains A and B have 21 and 30 amino acids (aa) residues respectively and are linked by two disulfide (S-S) bonds between cysteine residues. Insulin is transporter hormone in blood fluid controls and regulates response of certain cells or organs to level blood glucose (BGL) [5]. After a meal, insulin releases into blood stream. Insulin activate tissues cells to absorb blood glucose for growth, energy demands to avoid starvation. Insulin binding receptors on cell membranes, in response, G-transporter proteins are released from cell interior to cell membrane to carry and transport glucose from blood fluid into tissue.

Design and formulation of high-performance therapeutic DDS for oral administration of insulin is an advanced technology deals with human health [6]. Therapeutic insulin vials such as Lantus Glargine and Levemir Detemir are long acting insulin for human analogue. Glargine hormone is a four-polypeptide protein contains 10–50 amino acid residues regulates uptake BGL by human cells. It is a transporter protein binds and transport glucose molecules in blood fluid or through cell membrane [7, 8].

Diabetes Miletus (DM) is a chronic epidemic metabolic disease negatively impact many people especially in undeveloped countries. DM causes microvascular complications such as retino-, nephron-, neuro-pathy and diabetic foot disorders. Conventional therapeutic subcutaneous injected insulin for 1, 2 DM: T1DM and T2DM showed many drawbacks. Patients are non-compliance to insulin vial that may cause discomfort, pain and local infections [8].

Oral insulin administration mimic pharmacokinetics of endogenous insulin and improved patient acceptability. Sub-micron sized pH-sensitive, bio-degradable,-and compatible insulin carriers include: nano-, micro-particles and liposomes; layer-by-layer coating, self-polymerized shells and nanocomposites. However, these formulations have various challenges such broad size distribution, unreproducible uncontrolled initial burst insulin release, excessive hypoglycemia and immunological response [9]. This technology needs optimization of physiochemical and pharmacokinetic of insulin carriers.

The reported oral insulin medications included: Microspheres of poly(acrylic acid)-coated chitosan (CT)-5-F-uracil and Na taurocholate absorption enhancer [10, 11]. Insulin@alginate microspheres contain Ca(II) ions showed insulin leakage before target release [12]. Insulin@ β-cyclodextrin emulsions produced a dose-dependent hypoglycemia 13]. insulin@alginate chitosan microspheres had good broad release rate in response to pH but have broad size distribution and multi dispersity index [12]. Insulin carriers poly-(anhydride, p-(carboxyethylformamido)-benzoic anhydride] contain toxic chemicals [14]. Microemulsion insulin@Eudragit (L100, S100) showed strong adherence to GIT [15].

Anionic polymers blend methyl acrylamide (MAA) and MAA-methyl ester resisted enzymatic degradation causing insulin released at high pH [16]. Microspheres Insulin@-OH-propyl-methyl-cellulose acetate succinate efficiently encapsulated insulin (An aprotinin enzyme inhibitor, absorption enhancer Na-glycocholate) showed burst insulin release. Eudragit and polylactic acid microspheres had low insulin entrapment efficiency and enzymatically degraded. Trimethyl-CT facilitated insulin absorption in synergism with Eudragit clay [17]. For insulin@ Eudragit microspheres, Mg-stearate lubricant optimize emulsion droplet size [17].

Hydroxy propyl methylcellulose phthalate incorporated Na N-(8-[2-OH-benzoyl] amino) an absorption enhancer for insulin microspheres improved bioavailability [18]. Insulin@poly ester amide microspheres undergoes enzymatic hydrolysis ]19]. High Mw. chitosan microspheres increased insulin contact with duodenum and jejunum in diabetic rats [20] but showed low insulin absorption. This current study aims formulation of new oral administration insulin-DDS using natural polymer chitin and guar gum as inert biocompatible polymeric carriers. These novel formulations may overcome all reported obstacles in insulin oral administrations. This aim is also extending to evaluate invitro insulin release from these DDS in the simulated physiological fluids.

II.1. Materials

All chemical used in this study are all of analytical grades have high purity purchased from certified Co. of chemical products, and are used as received without further purifications,

SI.II.1. The novel sol-gel method for formulation of insulin drug delivery system (IDDS) is explored in the following section:

II.2. Methods

Two samples of IDDS: insulin@chitin and insulin@chitin-g-GG nanocomposites are prepared in 250 mL three neck cell following sol-gel method [21]. A weight 2.0 g chitin or (1g chitin + 1g GG) and traces amounts of the following chemical agents are used. Ammonium persulphate an initiator, Glutaraldehyde a cross linker, Fig.SI.2 [21], sodium dodecyl sulphate (SDS) as a catalyst, Table 1.

Table 1

The chemical constituents of IDDS
Physical characteristic of DDS			Insulin	Chitin	GG	Glutaraldehyde	NH₄₂S₂O₈	SDS
Formulation	Color	%Yield	Wt.(g)
Insulin@chitin	Pale yellow	86.1	3 mL	2	-	0.1	0.05	0.03
Insulin@chitin-g-GG	Dark yellow	75.2	3 mL	1	1	0.1	0.05	0.03

The specified weights of these materials are added to 40 mL double distilled water. Alkaline pH 8 is adjusted using 0.1M NaOH. All preparations are conducted at constant agitation 50 rpm. Reaction vessel is refluxed for three hrs. under inert nitrogen, flow rate 10 mL.min^− 1. The flask containing the reaction product is left to cool to the room temperature. The product precipitate is filtered under suction and it is washed three times by double distilled water to remove any unreacted homopolymer. The composite is grinded into nanoscale using mortar and pestle, followed by packing in sealed vials for characterization as discussed in, SI, section II.2. The details of monitoring insulin-release from IDDS is followed by spectrophotometric and electrochemical techniques, SI. section II.3.

Figure 1a, b showed FTIR spectra of insulin@chitin and insulin @chitin-guar gum respectively [30].

The distinctive FTIR spectra of both insulin@chitin and insulin@chitin-GG nanocomposites confirmed the modification of chemical structure of chitin on grafting by GG. Insulin@chitin-GG nanocomposite showed both functional groups of chitin (CH₂OH, CH₃CONH-) in addition to NH₂ and COOH of insulin. All the assigned FTIR vibrational bands at the characteristic wavenumber, (\(\stackrel{-}{\upsilon },\)cm^− 1) confirmed successful intercalation between chitin and GG polymeric matrix, Table SI.1. The SEM micrographs of native insulin and chitin as well as insulin@chitin and insulin@chitin-g-guar gum are shown in Figs. 2 (a, b) and 3 (a, b) at magnification 45000x.

The layered microstructure of insulin polypeptide is attributed to the interconnected polypeptide chains by: disulphide bond, hydrogen bond between amino (NH₂) and carboxylic (COOH) group of cysteine amino acids (aa) residues [6].

Bright fine spherical Chitin nanoparticles (NPs) are regularly and homogeneously distributed on insulin polypeptide chains. Morpholgy of chitin and insulin are maintained in both IDDS.The copolymer is a smoother polymeric carrier for insulin therapy. Insulin chains are more firmely linked to chitin NPs as carbonyl group in chitin acetamide (NHCOCH₃) is broken in alkaline pH of reaction media for binding NH₂ group of insulin molecules.

In insulin@chitin-g-GG, acloholic CH₂OH group of chitin form intermolecular hydrogen bonding H.B. with guar gum [6]. Some aggregations are also attibuted due to intra-molecular H-B. within GG chaines. No aggregation or agglomeration are observed in both IDDS.

Powder XRD patterns of DDS sample are show in Fig. 4a, b

PXRD pattern of insulin@chitin showed semi-crystalline structure characterizing native chitin. Insulin@chitin-g-GG has both crystalline and amorphous domains [31]. Grafting chitin by GG declined crystallinity of chitin, improve proteolytic stability and membrane permeability of this IDDS. The higher crystalline insulin@chitin indicated that N atom of (NH) group of chitin is strongly binding to O atom of COOH of insulin in a bidentate chelating mode. Binding GG to both insulin and chitin involved H.B. and Van der Waals interaction. Figures 5 (a, b) and 6 (a, b) showed TGA and DTA thermograms respectively.

These Figs.showed that insulin@chitin exhibited better thermal stability compared to Insulin@chitin-g-GG. The comparative thermal parameters of the tested samples are collected in Table 2 [32].

Table 2

Thermal data of insulin@chitin and Insulin@chitin-g-GG
Sample	Steps	t, °C	Wt. loss%	Residue%	DTA, peaks t, °C
Insulin@chitin	I II III	36.4–74.8 74.8-230.4 230.4–696.0	3.45 18.30 27.74	66.86	86.3-140.8 (endo, intense) 248.4-292.7 (exo weak) 295.5-381.1 (exo weak)
Insulin@chitin-g-GG	I II	36.3–376.0 376.0-700.1	59.78 21.53	18.59	276.5-402.1 (exo, broad& intense), 447.4-508.2 ( exo, broad& intense)

Insulin@chitin thermally decomposed in three steps, wt. loss % 3.45, 18.30, 27.74, and 66.86 residue. Insulin@chitin-g-GG decomposed in two steps, wt. loss% (59.78%, 21.53% (higher than Insulin@chitin) and only 18.59 residue. Insulin@chitin exhibited only one endothermic peak at temperature range: 86.3^oC-140.8^oC due to the strong binding between functional groups of chitin and insulin. Insulin@chitin-g-GG exhibited two broad and intense exothermic peaks due to bond formation between various decomposed carboneous polymeric residues species [33].

Figure 7 showed UV-Vis.absorption bands for the known concentrations of insulin used in construction of its calibration curve, Fig.SI.3.

Applying the least square method on absorbance-concentration plot, good straight line is obtained with slope equals molar extension coefficient, ε 0.005). Insulin concentration is determined using Beers Lambert law. The high correlation coefficient R² 0.9984 confirmed the validity of UV-Vis. spectroscopy for monitoring invitro-insulin release. Fig. SI.3 (a-c) showed UV-Vis. spectra for insulin release from insuline@chitin-g-guar gum [27].

Figure 8a, b represented insulin release form insulin@chitin at pH 7.4, insulin@chitin-g-GG at pH 1.2, 6.8, 7.4 respectively.

Insulin release at pH 7.4 from insulin@chitin give 5 ppm negligible maximum concentration. Insulin release from this IDDS at pH 1.2 and 6.8 cannot be followed spectrophotometrically. Insulin@chitin-g-GG showed higher concentration (10 ppm,16 ppm and 18 ppm at pH 1.2, 7.4, and 6.8) respectively of released insulin than insulin@chitin. The initial rapid burst Insulin-release is attributed to the fast detachment of encapsulated insulin which adhered to surface of polymer shell in response to pressure of the media [34]. Burst release at about 15 min. is in agreement with half life time of insulin pharmacokinetic in blood stream as few min. [35]. Insulin@chitin-GG showed controllable insulin release that is limited at 76.2%, 80.7% and 67.2% at pH 1.2, 6.8 and 7.4 respectively. Insulin released data from insulin@chitin-g-GG are linearly fit to different kinetic models, SI.III.3 applying least square method in linear regression analysis. The suitable kinetic model for release kinetic is selected to have highest correlation coefficient, R² ≥ 0.95 between the dependent parameter y and the independent parameter x in the empirical equation of each kinetic model [29].

Table 3 collected R² obtained from the linear fitting to release data of insulin to the tested kinetic models.

Table 3

Correlation coefficient, R² for linear fitting of release data to kinetic models
pH	Run	Zero order		Pseudo 1^o		Pseudo 2^o	Elovich	Higuchi	Ritger-Peppas
					Correlation coefficient, R²
1.2	1		0.7259		0.8845	0.9965	0.943	0.8452	0.897
	2		0.8338		0.974	0.9994	0.9857	0.9261	0.9609
6.8	1		0.3691		0.3611	0.9959	0.5906	0.4794	0.6197
	2		0.3977		0.3821	0.9618	0.6574	0.5293	0.6915
7.4	1		0.7466		0.7549	0.9975	0.8645	0.8098	0.8368
	2		0.7721		0.7889	0.9998	0.9652	0.8815	0.9432

Higuchi plot model showed low correlation coefficient, therefore the mechanism of insulin desorption from polymer matrix is not under diffusion control as suggested by this kinetic model [38]. The linear fitting of insulin release to pseudo second order kinetic model gave the highest value of R² in the range (0.9618–0.9998) [6], Fig. 9. Hence pseudo 2^o model is the suitable kinetic model for describing release kinetics of insulin from the prepared IDDS.

Figure 9 showed plot t/q versus t for pseudo second order kinetic. The linear regression equation, R² and the standard errors (S.D.) are collected in Table SI.3. The kinetic parameters k₂, q_e are obtained from the slope and the intercept of the obtained straight line and are collected in Table 4.

Table 4

kinetic parameter of pseudo-2^o model
pH	Run NO.	k₂, g/mg min	q_{e cal}., ppm	h, ppm. g^− 1 min^− 1	R²
1.2	Run1	0.0073	19.64	2.816	0.9965
1.2	Run2	0.0067	19.92	2.674	0.9994
6.8	Run1	0.0305	11.32	3.908	0.9959
6.8	Run2	0.0290	15.79	7.237	0.9618
7.4	Run1	0.0134	15.12	3.063	0.9975
7.4	Run2	0.0161	21.05	7.133	0.9998

The best fitting of data of insulin release to Pseudo 2^o order model indicated that of insulin@chitin-g-GG is heterogeneous solid surface has active sites of different energy [40]. Each insulin molecule is release from two adsorption sites. The rate constant k₂ for insulin release followed the order:

pH 6.8 > 7.4 > 1.2

This trend is confirmed by impedance measurements expressed as Nyquist plots, Fig. 10

Charge transfer resistance (R_ct) equals diameter of semicircle. The smaller R_ct for insulin @chitin-GG at all pH indicated that low concentration of insulin released due to tight linkage to chitin. Correspondingly, few insulin molecules are liberated from insulin@chitin microcapsule to adsorb on surface of working electrode by heteroatoms of functional groups. DDS polarized by applied AC current field, the liberated insulin from microcapsule adsorb on surface working electrode increased charge transfer resistance over electrode surface [29].

Rate of insulin release is retarded at pH 1.2, while there is a significant release at pH 6.8 and 7.4.

For insulin @chitin-GG, slow insulin release at low pH indicating insulin protection against aggressive HCl in GIT environment (pH 1–3). Hence increases transit time till passage into small intestine for enhanced absorption in blood stream. Chitin is mucoadhesive polymeric carrier retard insulin degradation, hence reduces insulin loss in gastric acid and intestinal environment. At pH 6.8 of body fluids, the absorption enhancers in mosaic layer and enterocytes connected by tight junction improved release and absorption of insulin.

This result also confirmed that insulin release is pH dependent, with a higher release rate at colon pH (6.8), where insulin-NPs connections become weak and insulin is released without any protein degradation. Similar pH response, such as retention of insulin within polymer matrix at acidic pH and significant release at higher pH are also observed in insulin-loaded alginate microspheres [12, 27].

Applied AC electric signal potential on various applied frequency polarized electron clouds and changed the dielectric properties in atoms of IDDS generating electron-hole pair and current flow. The amphoteric Zwitter ionic amino of insulin confirmed polarizability of insulin monomers [6, 40]. Capacitive impedance plots confirmed the macroscopic dielectric properties and polarizability of IDDS as well charge separation and phase transition [41, 42]. The mucoadhesive GG enhanced insulin release in a controlled sustained manner via improving hydrophilicity, charge density, pH-sensitivity and response [40].

Impedance plots confirmed pH effect on insulin release from DDS in simulated physiological fluids. Release insulin from microcapsule core via swelling of polymeric shell by aqueous buffer solution. Relaxation of released insulin appear as one time constant.

Impedance results satisfactory agreed UV-Vis. Spectroscopy in the fact that insulin@chitin is very difficult to release insulin. This finding suggested the validation of impedance spectroscopy is novel technique for qualitative determination of release insulin in physiological body fluids.

The better performance of insulin @chitin-g-GG in insulin release is interpreted in terms of swelling and porosity [22, 23], Table 5.

Table 5

Porosity and swelling% of insulin DDS
Sample	Chitin	Guar gum	Insulin @chitin	Insulin @chitin-g-guar gum
Porosity	503 ± 20	723 ± 37	63.37 ± 20	47.37 ± 20
Swelling%	3100	2900	980	650

The largest swelling percentage belonged to chitin. GG the greatest ability to form intramolecular H.B. (four H.B. for each monomeric unit) forming 3D network structure but its high rheology limited water absorption. Although the swelling of insulin @chitin exceeded that of insulin @chitin-g-GG, however the strong linkage between semi crystalline chitin and insulin retard insulin release [22].

The largest porosity of GG is attributed strong intramolecular H.B. The lower porosity of insulin @chitin-g-GG although having better insulin release suggested high cross linking on this DDS increased insulin loading capacity. Insulin loaded by both penetration into pores and binding multiple functional groups. However, chitin-g-GG copolymer is loaded by more insulin peptide chains decreased porosity of DDS than that of insulin@chitin [23].

Swelling of DDS in aqueous physiological body fluid enabled absorption of body fluid, increase interior surface accessible for body fluid. Insulin @chitin-g-GG has hydrogel character behave as low viscus liquid with rapid phase transition. Grafted chitin is loaded by more insulin than native chitin due to multiple functional groups [23].

Amino acid test from IDDS along 8 hrs. immersion in phosphate buffer saline (BFS), Table 6 confirmed more peptides and aa sequence from insulin@chitin-g-GG [22, 23].

Table 6

Peptides and amino acid sequence for insulin drug delivery systems
Variable	Insulin DDS
Variable	Insulin@chitin			Insulin@chitin-g-GG
C _mg/mL	1	3	8	1	3	8
Peptide	0.39	0.43	0.50	0.61	1.46	2.27
aa	0.05	0.51	0.9	0.071	0.69	1.04

Peptide and aa produced by degradation of insulin by ninhydrin as insulin monomer is a biological polypeptide protein hormone; Mw. 5.800 k Da.. The aa are linked by peptide bond via removal water molecule [42] giving dipeptide that link a third aa yield tripeptide. Repeated peptide bond formation gives specific aa sequence of insulin [22], especially sulfur-containing aa cysteine and methionine.

Insulin exists in solution as monomers, dimers, tetramers, and hexamers due to ions-, and solvent interaction. The aa monomers of insulin are nonpolar hydrophobic, polar, neutral, basic, and acidic. These aa differ in NH₂ position to COOH group [23]. Insulin has primary protein structure (chains of aa sequence and S-S linkage) has neither α-helix pleated sheet nor 3D conformation in addition to the absence of any associated two or more chains.

The formulated oral administration nanocomposites of IDDS showed insulin loading efficiency and permeability to biological fluids facilitate insulin absorption and bioavailability.

Figure 11 showed that both IDDS have the same particle size distribution between nanoscale microscale (70 nm-1000nm) up to 1 µm. confirmed successful insulin loading on polymeric carrier.

Particle size distribution is slightly modified at µm region for insulin@chtin-g-GG due to increased surface area by additional functional groups [26].

Negative zeta potential, Fig. 12 for both IDDS confirmed stability against coagulation and enabled outstanding to storage temperatures, mechanical shear force, etc. to ensure sustained insulin release [26].

Chitin-g-GG is ether copolymeric hydrophilic hydrogel arranged in a block, random or alternating configuration throughout polymeric chains network, or interpenetrating polymeric hydrogel or polymer blends [40–43].

The traces amount of catalyst, initiator and cross linkers used in preparation of IDDS are all below the toxicity limits. Both insulin@chitin and insulin@chitin-g-GG nanocomposites could not show neuroinflammation or alteration autophagy metabolic process [44, 45].

The natural polymers chitin or chitin grafted guar gum are used as inert biodegradable drug carriers in formulation of two bioactive DDS for oral administration of insulin vials. Insulin hormone is loaded on the native chitin or its copolymer by using simple sol-gel method. GG increased the numbers of functional groups on copolymer. Insulin-chains of amino acid sequence bounded to chitin decreased rigidity of chitin chains. Insulin@chitin has depth pores, while the smoother insulin@chitin-g-GG multiple functional groups and little pores. Polymeric chains of chitin and GG are extensively cross linked via intermolecular H.B. Insulin@chitin is semicrystalline with well defined deep tunable pores. Insulin@chitin-g-GG has both crystalline and amorphous features. Chitin strongly binding insulin by NH-, OH group and O atom of COOH group in insulin. The coplymer linked insulin via H.B. and Van der Waals interaction. Insulin chitin is more thermally stable than insulin@chitin-g-GG. Insulin@chitin-g-GG showed better sustained insulin release than insulin@chitin. The best better linear fit of release data to pseudo 2^oorder indicated that chitin-g-GG is heterogeneous solid surface and each insulin molecule occupied two active sites. Targeting insulin-release at physiological temperature 37 ± 0.5ºC followed the order: pH 6.8 > pH 7.4 > pH 1.2. This trend is confirmed by impedance measurements. The low insulin release rate at pH 1.2 indicated its protection against acidic- or enzymatic hydrolysis in GIT for targeting sustained absorption in blood stream to regulate BGL. The physicochemical characteristics of insulin@chitin-g-GG and insulin@chitin porosity, swelling ratio, peptide &amino acid sequence, zeta potential, particle size distribution and pseudo 2^o kinetics reflect that: Insulin loaded on chitin by chemisorbed monolayer and loaded on chitin-g-GG by chemi-, and phyi-sorption. The latter adsorption mechanism is caused by electrostatic bonding to multiple functional groups of copolymers facilitated insulin release rate. Chitin surface act as electron acceptor Lewis acid. Chitin-g-GG copolymer acted as both Lewis and Bronsted acid (gives proton or accept OH⁻). Loading capacity of insulin may be higher than that of chitin-g-GG. However, the strong irreversible chemical bonding to insulin retard release rate.

Supporting Information Summary

Supporting information includes all remaining data and materials of this current study.

Ethics approval and consent to participate

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Availability of data and materials

All data and materials of this study are available in the manuscript and supplementary information.

Competing interests

Authors have declared that no competing interests exist. The products used for this research are commonly and predominantly use products in our area of research and country. There is no conflict of interest between the authors and producers of the products because authors do not intend to use these products as an avenue for any litigation but for the advancement of knowledge.

Funding

Open access funding provided by Science Technology&Innovation Funding Authority (STDF) in cooperation with The Egyptian Knowledge Bank (EKB). Funding was provided by Alexandria University.

Author contribution

H. A. Fetouh, E.E. A.Aleem, N. H. Mohammed, A. M. Ismail conceived the study; A. M. Ismail and H. A. Fetouh suggested the research point; A.Aleem help to explore null toxicity of trace chemicals used; N. H. Mohammed carried out spectrophotometric measurments under supervision of A. M. Ismail; H.A.Fetouh performed impedance measurements; A. M. Ismail and H. A. Fetouh wrote the first draft of the manuscript

Acknowledgements

Authors acknowledged Alexandria University for faciliatating funding provided by Science Technology & Innovation Funding Authority (STDF) in cooperation with The Egyptian Knowledge Bank (EKB).

Authors' information

Authors

Howida A.Fetouh Tel.:+201203166565. E-mail address: [email protected]

Engy E. Abdel Aleem Tel.:+201002804461. E-mail address: [email protected]

Najiyah H.Mohammed Tel.:+201061156198.E-mail address: [email protected]

Amel M.Ismail Tel.:+201090010143. E-mail address: [email protected]

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New advanced approach for processing natural polymers chitin and guar gum for developing new bioactive biocompatible drug delivery systems for oral administration of insulin

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I. Introduction

Ii. Materials And Methods

II.1. Materials

II.2. Methods

Iii. Results And Discussion

Iv. Conclusion

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